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CombiFlash Rf User Manual with Rf150

1. 5 4 1 Run Control Buttons e Play The Play button starts the run During the run other run control buttons are active e Pause The Pause button holds the B at the current value while the system continues to operate sometimes called an isocratic hold Note that pausing the run extends the run length While in the Paused state you 5 9 CombiFlash Rf User Manual can resume the run by clicking the Play button or stop the run by clicking the Stop button If you resume the run the system continues with the gradient curve at the B when the system was paused e Stop The Stop button suspends the entire run Unlike the Paused state the pump peak detection and fraction collection will not operate While stopped you can end the run by clicking the Rewind button resume the run by clicking the Play button or skip the rest of the run sequence by clicking the Fast Forward button CAUTION Rf200 and Rf200i only When the Stop button is pressed solvent remains in the tubing from the injection valve to the fraction collector s drop former Follow the steps listed below considering your instrument s configuration Solid Sample Cartridge with air purge enabled If you suspend the run with the Stop button do not remove the column and or solid sample cartridge without first pressing the Fast Forward button to purge solvents from all lines If you stop the run because your peak of interest has been collecte
2. Comments 4 15 CombiFlash Rf User Manual CombiFlash Rf User Manual Section 5 Operation This section provides abbreviated operating instructions for the CombiFlash Rf systems For complete instructions refer to Help menu option from PeakTrak s main menu 5 1 Sample Preparation Before starting a run consider how the sample will be introduced to the column media This section discusses three methods liquid injection solid sample cartridges and preloading the sample on the column 5 1 1 Liquid Sample Injection If the sample is soluble in the starting mobile phase it can be prepared as a solution and injected onto the column when prompted during the purification run 5 1 2 Solid Samples Some compounds are not soluble in solvents that are compatible with the chromatography In addition the compounds may have very limited solubility in any solvent resulting in sample volumes that are impractical for good chromatography The answer in this case is solid sample introduction Here the reaction mixture or dissolved sample is mixed with the same media in the chromatography column Some samples of this type may be simply dissolved and placed into a prefilled cartridge You can then use the cartridge immediately or dry it before placing it on the system Other samples may need more care Typically you would create a mixture of 20 sample load to media w w Then dry the mixture under
3. Figure 5 3 Align and push cartridge into the cap and then rotate the cartridge to secure it 5 4 Section 5 Operation 5 1 3 Preloading on Column You can load the reaction mixture or sample directly onto the column sometimes called direct or dry loading M Note If using this method use care to ensure that a column equilibration does not wash away the sample Bypass the column equilibration at run time by selecting Preload on column or No Equilibration for Rf 150 systems as the sample Loading Type Refer to Section 5 3 for more details 5 2 Loading a RediSep Rf Column To load a RediSep Rf column NWARNING The CombiFlash Rf has redundant safety devices to limit pressure to less than 210 psi 1448 kPa RediSep columns smaller than 100 g are CE certified using standard IEC61010 1 for use on the CombiF ash Rf RediSep columns larger than 100 g meet Pressure Vessel Directive 97 23 EC Teledyne Isco strongly recommends against the use of columns rated less than 210 psi 1448 kPa For best results always use RediSep Rf columns The system will not automatically detect other columns including non Rf RediSep columns The system also limits the maximum operating pressure to 50 psi when the column is not detected 1 Select a RediSep Rf column The on line help includes a column selection guide that can assist you in selecting a stationary phase media and column size 2 Raise the injection valve c
4. RediSep Select the column that will be used for the run See also REDISEP RF COLUMN SELECTION GUIDE ON LINEJ Loading Type Select the type of injection Start Tube Select Next Tube to begin at the next available tube in the current rack or Next Rack to begin with the first tube in the next rack 6 3 5 2 Gradient Plot Area The Gradient Plot Area is the same as the one displayed on the tabs for each column position The display shows the current channel The top corner displays the current system pressure 6 3 5 3 Run Control Buttons The Run Control Buttons allow you to control the operation of the system 6 3 6 Files The Files window is modal That is its function and features change according to the command used to open the window Menu commands such as FILE gt OPEN 6 2 1 2 and SAVE METHOD AS 6 2 1 6 or clicking Open and Save As buttons will open this window Use this window to browse the system s files and folders The following controls appear on the window Current Path The top left corner of the window displays the path current folder As you browse through the files the path will update as you go You can click the folder names to return to upper folder levels File and Folder Operation buttons Copy Click this button to copy a highlighted file to the system s clipboard memory Paste Click this button to paste a file from the clipboard memory If the file already exist
5. 3 3 Connect Power Ensure that the On Standby switch below the touch screen panel is in the Standby position Then use the supplied IEC power cord to connect the CombiFlash Rf to mains power WARNING Mains power must meet the voltage frequency and amperage requirements listed on the serial number label Z WARNING As long as the AC mains power cord is connected power is inside the unit The mains power cord is the disconnect device Position the Rf 150 system so that the power cord can be unplugged or use a power strip where the plug can quickly be removed from the outlet in the event of an emergency CombiFlash Rf User Manual 3 4 Connect Solvent Lines A WARNING Risk of fire ignited by electrostatic discharge Never substitute the black tubing on CombiFlash systems The black tubing P N 023 0503 06 is conductive This tubing is required to dissipate static electricity A CAUTION To prevent damage or premature wear to the pump and internal valves clean solvent should be used The solvent should not contain any dissolved solids The Combiflash Rf 150 system has two solvent inlet ports on the back panel one A solvent and one B solvent Figure 3 2 Solvent Inlet Ports Figure 3 2 CombiFlash Rf150 solvent inlet ports The system requires at least one A and one B solvent to form a binary gradient It assumes that the B solvent is stronger than A When using th
6. You can reset the method to Teledyne Isco recommended defaults While viewing the default method file for a selected column size and type click the Reset file to Isco defaults 6 43 CombiFlash Rf User Manual button The settings will revert to the original factory settings Click the Save button to save the settings for future use 6 4 5 Defining a Gradient The simplest way to change the gradient is to click and drag the inflection points that define the shape of the gradient Add a point by clicking the Insert button then click the gradient curve to add a point You can then drag the new point to the desired location Delete a point by clicking the Delete button and then clicking the undesired inflection point The above methods work in the MAIN WINDOW 6 3 1 and METHOD EDITOR 6 3 2 window Alternatively you can modify add insert and delete points using a tabular view on the Method Editor window Click the Edit Gradient button to open this view then use the controls to modify the settings Click the Edit Gradient button again to close the table view The system can also optimize a gradient from your TLC results To use this feature select a single solvent system and run two TLC plates each with a different concentration of solvent For example run one plate at 1 1 hexane ethyl acetate the other at 1 3 hexane ethyl acetate If the resulting Rf values are between 0 2 and 0 8 you can enter the TLC data in the G
7. 1 3 Operating Overview The Combiflash Rf system is equipped with a touch screen display for local control The system supports TCP IP communication This allows direct control of the system by a Windows computer via a cross over cable between Ethernet ports of the CombiFlash Rf system and the computer The Windows computer must have Microsoft Internet Explorer 6 or newer TCP IP communication also allows remote control of the system via an established network Remote controlling devices on the network can be a Windows PC or an Apple mobile digital device iPod Touch iPhone and iPad M Note Teledyne Isco recommends that you obtain assistance from your Information Technology department before attempting direct or network connections 1 3 1 Multiple Control Possibilities The system can be accessed from the built in touch panel and up to ten network devices The touch panel shares control with all connected devices The system performs the most recent command from any control input 1 3 2 File Storage To support operation from a variety of direct and network connections the software and all files are stored in the CombiFlash Rf on an internal hard drive This ensures that your compound purification methods and run history files can be viewed from any connection Optionally individual run history files may be saved to a USB flash drive and the hard drive or a networked computer and all run history files can be saved to a networ
8. 465 3799 Laboratory Equipment for Light Industrial Commercial Environments Combiflash RF75 2008 EN 61010 1 2 Edition Safety Requirements for Electrical Equipment for Measurement Control and Laboratory Use EN 61326 2006 EMC Requirements for Electrical Equipment for Measurement Control and Laboratory Use Standard Description Severity Applied Performance Criteria EN61000 4 2 Electrostatic Discharge Level 2 4kV contact discharge A Level 3 8kV air discharge EN61000 4 3 Radiated RF Immunity 80 Mhz to 2 7Ghz 80 AM at 1kHz A Level 2 3V m EN61000 4 4 Electrical Fast Transient Level 2 1kV on AC lines EN61000 4 5 Surge on AC Lines Level 2 2kV common mode Level 2 1KV differential mode EN61000 4 6 Conducted RF on AC lines 150 kHz to 80 MHz A Level 1 1V rms 80 modulated EN61000 4 11 Voltage Dips 1 10 25 cycles 100 40 70 A Voltage Interruption 250 cycles 100 drop c CISPR11 RF Emissions Group 1 Class A Industrial Scientific and EN 55011 Medical Equipment EN61000 3 2 3 3 Harmonic and Flicker The undersigned hereby declares that the design of the equipment specified above conforms to the above Directive s and Standards as of September 5 2008 USA Representative odis William Foster Vice President of Engineering ZAN TELEDYNE ISCO 60 5232 506 A Teledyne Technologies Company DECLARATION OF
9. 7 10 Section 7 Remote Interfaces Local Area Connection Properties E Client for Microsoft Networks File and Printer Sharing for Microsoft Networks Y 3Com BCAITDI DMI TDI Internet Protocol TCP IP Properties 192 168 1 200 Figure 7 7 Internet Protocol Properties Window 7 11 CombiFlash Rf User Manual 7 2 5 Completing the Direct Connection 1 Connect the PC to the Combiflash Rf system using the cross over cable 2 Turn the system s On Standby switch to ON Wait for the PeakTrak operating screen to appear on the Combiflash Rf system then start the PC 3 Start Internet Explorer and type the factory default address http 192 168 1 51 The browser will display the PeakTrak screen The PeakTrak screen indicates that the direct connection is working The CombiFlash Rf system and the PC are now configured for communication through a direct connection Refer to PeakTrak s on line help for operating instructions Abbreviated instructions can be found in Section 5 of this guide 7 3 Direct Printing The CombiFlash Rf system supports direct printing to a USB printer The printer must support postscript To set up the printer 1 Connect the printer with the USB cable to the USB port on the back panel of the system 2 Select the Tools Configuration menu command 3 Select the Network Configuration tab 4 Select the Postscript option for the Printer Type 5 For the Connection URL
10. 8 5 8 Monthly Flow Cell Cleaning 8 10 8 6 Injection Valve Maintenance 8 10 8 7 Electrical Troubleshooting 8 11 vi CombiFlash Rf User Manual Section 1 Introduction 1 1 Documentation Overview This Installation Guide provides e Safety Information Unpacking instructions nstallation instructions including connections with a Windows computer Certification and warranty information Once you are operating the CombiFlash Rf you may refer to the Help menu for operating instructions M Note This Installation Guide will use the term Combi Flash Rf when the context applies to all systems This installation guide will refer to the specific model name when the discussion is limited to a particular model or configuration 1 2 Product Overview The Teledyne Isco CombiFlash Rf flash chromatography system gives you high productivity automation programmable gradients UV detection and peak separation and automatic detection of columns and collection tube racks with the exception of the Rf150 Its small size makes it a great personal system and well suited for operation within chemical hoods and other limited indoor spaces Z WARNING Avoid hazardous practices If you use this instrument in any way not specified in this manual the protection provided by the instrument may be impaired this may increase your risk of injury CombiFlash Rf User Manual
11. Flow Cell Pathlength 0 1 mm 25 UV Detection and Monitor Wavelength Selectable from 200 to 360 nm range Optional 200 to 780 nm UV Vis detector Wavelength Accuracy 5 nm Gas Inlet Pressure 60 to 70 psig Gas Consumption lt 2 5 SLPM Spray Chamber Temperature Setting range 10 to 60 C limited to minimum of 5 C below ambient Drift Tube Temperature Setting range 30 to 90 C Must be 5 C above spray chamber temperature Minimum temperature is 5 C above ambient Maximum temperature is 60 C above ambient 1 9 CombiFlash Rf User Manual Table 1 2 CombiFlash Rf 200i Specifications Continued Split Flow Rate 0 75 mL min 10 Fraction Accuracy 2mL flow rate 60 Electrical Safety per EN 61010 1 Pollution Degree 2 Installation Category II Maximum Altitude 2000 meters Note 1 All specifications are subject to change Note 2 For columns undetected by RFID the max pressure is limited to 50 psi Section 1 Introduction Table 1 3 CombiFlash Rf 4x System Module Dimensions 1 Specifications 61 cm 24 x21 6cm 8 5 x43 2 cm 17 HxWxD Module Weight 13 6 kg 30 Ibs Overall Dimensions 61 cm 24 x57 6cm 22 6 x43 cm 17 HxWxD Overall Weight 40 8 kg 90 Ibs Power Options Line Frequency Ambient Temperature Input voltage range from 100 to 240 VAC 50 60 Hz 2 5 Amps system maximum 0 5 amps
12. M Note When using higher density solvents such as dichloromethane DCM ensure that the waste container is no more than 3 feet below the supply solvent levels Placing solvent containers above the system and waste containers on the floor may cause the internal check valves to open and allow solvent to flow through the system when in the standby state 2 6 External Gas 2 6 1 CombiFlash Rf 200 and Rf 200i The CombiFlash Rf 200 and Rf 200i systems use an internal air pump to provide compressed air for solvent and waste level sensing Optionally the system can deliver a compressed gas Inert or other gases may be more suitable than air for some applications If ambient air is compatible with your solvents and waste simply leave the Optional External Gas port open If you want to use a gas instead of air connect user supplied lg I D tubing to the Optional External Gas port Connect the other end to the user supplied compressed gas The external 2 8 Section 2 Preparation Rf 200 and Rf 200i Systems compressed gas should be dry filtered and regulated to 2 5 psi 0 14 0 34 atm 13 8 34 5 KPa 2 6 2 CombiFlash Rf 200i Only ELSD operation requires a carrier gas To connect the carrier gas to the CombiFlash Rf 200i 1 Locate the 4s inch O D FEP tubing part number 023 0503 02 from the accessory package 2 Push one end of the tubing into the Nitrogen Inlet port on the back of the system The tubing should be full
13. The Rf150 cannot read the RFID tag 7 Sample Injection Port Luer type fitting to accept the sample though either a solid load sample cartridge shown or a liquid injection using a syringe or similar device 8 Injection Valve This six way valve is automatically positioned by the Combiflash Rf according to its current mode of operation Modes include column equilibration sample injection elution column flushing valve cleaning and system purging The injection valve is not available on the standard Rf 150 systems 9 Upper Column Mount Secures the column inlet 10 Lower Column Mount Secures the column outlet 11 Top Shelf Allows storage of accessories and solvent bottles 12 Column Expansion Ports These ports allow the system to add column channels with an optional module The 4x module option is not available for the Rf 150 systems Figure 1 6 CombiFlash Rf Features Side Section 1 Introduction Figure 1 7 CombiFlash Rf Features Back 13 Upper Drain Tube Liquids spilled on the top shelf are carried away through this tube to a user supplied container 14 External Detector All systems except the Rf 150 The system can accept an analog peak voltage signal from an external detector The system will process this analog signal to determine the presence of a peak using PeakTrak s slope or threshold algorithms See section 2 10 more information 15 Ethernet Port An 8P8C jack for a netw
14. enter usb dev usb lp0 6 Click OK to save the settings The CombiFlash Rf system will send a test page to the printer After successfully printing a test page the local printer will be available for printing using the File Print menu command or through the Automatically Print Report at End of Run option on the Tools Configuration User Preferences tab 7 12 CombiFlash Rf User Manual Section 8 Maintenance 8 1 System Standby and Shut Down During extended periods of inactivity you can place the system in Standby to conserve power To do so log off the system File gt Log out and place the On Standby switch in Standby When in the Standby state normal system operation is no longer available from the touch screen or remotely However some internal components are still powered WARNING As long as the AC mains power cord is connected power is inside the unit The mains power cord is the disconnect device Position the CombiFlash Rf system so that the power cord can be unplugged or use a power strip where the plug can quickly be removed from the outlet in the event of an emergency When you first place the system in Standby internal components continue to operate for almost one minute while performing file maintenance and preparing the system for possible power removal N CAUTION Removing the AC mains power cord before the file maintenance is complete might corrupt files on the internal hard driv
15. s internal hard drive 6 2 1 3 Manage Files Choose this command to open a window from which you can archive or delete method and run files from the current directory You can archive files by connecting a USB Flash drive and selecting the Copy or Move files to flash drive option To remove a file from the internal hard drive highlight the file or enter the filename and click the Delete button To remove multiple files click the Delete by Age button Then specify the age limit in months and click OK Use the Move and Delete options with caution as the files cannot be recovered from the internal hard drive once the action is complete 6 2 1 4 Set Data Path This opens the SET DATA PATH 6 3 7 window Use this window to specify the default storage folders for users These folders contain multiple METHOD FILE 6 1 2 and RUN FILE 6 1 3 resources created by system users See also USER MANAGEMENT 6 3 12 6 2 1 5 Save Method Choose this menu command to save the settings of the current method file to the system s internal storage The file will be saved under the current file name Note DEFAULT METHODS 6 1 2 1 are an exception and cannot be overwritten by the Save Method command If a default method is open and you attempt to save any modifications to 6 4 Section 6 User Help Reference that method the FILES 6 3 6 window appears so you can rename the method file This preserves the default method If you do wi
16. the Main window displays peak collection data as well as the settings used for that run The Main window dynamically changes to display the controls required for the current state of the system The top region of the Main window always displays a GRADIENT PLOT 6 3 1 1 area The bottom region of the window displays the RUN SETTINGS 6 3 1 2 RUN CONTROL BUTTONS 6 3 1 3 and PEAK COLLECTION BUTTONS 6 3 1 4 After a run the left side of the window changes to show the collection rack map or the method parameters in a PEAK COLLECTION DATA 6 3 1 5 pane Note The main window is expanded for CombiFlash Rf 4x systems The main window the displays the settings and views in four tabs one for each column channel A fifth tab BATCH 6 3 5 contains the controls to run sequential separations 6 3 1 1 Gradient Plot The gradient plot area depicts the current gradient that will be used for the run The X axis of the plot displays the run time in minutes or COLUMN VOLUMES oN LINE The X axis scale can be adjusted by typing or selecting the Run Length The left Y axis displays absorbance units and the right Y axis indicates the percentage of solvent B By default PeakTrak automatically scales the left Y axis to best display the 6 10 Section 6 User Help Reference absorbance trace PeakTrak also sets the right Y axis to 100 You can override the Y axes scales by clicking the Format Graph button and setting the desired values
17. 13 3 10 2 General Settings 005 3 13 3 10 3 Set Date Time 00000005 3 13 3 10 4 Vapor Limit Optional 3 13 3 10 5 Set Default Tube Volumes 3 14 3 10 6 User Preferences 00000 ee eee 3 14 3 10 7 Restart the system 005 3 17 3 11 Prime the Solvent Lines 3 18 3 12 System Verification 0 0000 RR KK 3 19 3 13 Installation Qualification Checklist 3 20 4 Preparation Optional 4x Module 4 1 Unpacking the Unit sees 4 1 4 2 Instrument Location 0 00 cee eee 4 2 4 8 Connect Power cece eee eee 4 2 4 4 Connect Solvent Lines 0 00000 4 3 4 5 Connect Waste Lines RR KK 4 4 4 6 Control Connection 0 0c RR RR KK 4 11 4 7 Position the System RR KK 4 11 4 8 Remove Shipping Hardware 4 11 4 9 Turn on Power kk KK KK eee ees 4 12 4 10 Prime the Solvent Lines 4 13 4 11 System Verification 0 20000 eee eee 4 14 4 12 Installation Qualification Checklist 4 15 5 Operation 5 1 Sample Preparation 000 KK 5 1 5 1 1 Liquid Sample Injection 5 1 5 1 2 Solid Samples RR KK 5 1 5 1 8 Preloading on Column 5 5 Table of Contents 5 2 Loading a RediSep Rf Column 5 3
18. 3 4 command instead 6 14 Section 6 User Help Reference Save As Click this button to open the FILES 6 3 6 window From this window you can rename the current method and save it on the system s internal storage Exit Click this button to close the Method Editor and return to the MAIN WINDOW 6 3 1 Gradient Optimizer Click this button to open the GRADIENT OPTIMIZER 6 3 3 window This feature produces an optimized gradient optimized based on your TLC retention factor data Column Data Rf200 Rf200i and RfAX Click this button to open the COLUMN DATA 6 3 4 window This window reports information about the column installed in the system Time to CV CV to Time Click this button to toggle the Run Length X axis units between minutes and COLUMN VOLUMES oN LINE You can configure Peak Trak s default Run Length units by changing the CONFIGURATION 6 3 10 settings 6 3 2 2 Run Settings RediSep Column Rf200 Rf200i and Rf4X The RediSep column media and size are detected by the system and reported here The column should match the one that will be used for the run You may override the detected column size by selecting a different size See also REDISEP RF COLUMN SELECTION GUIDE oN LiNE and ABOUT REDISEP RF COLUMN DETECTION ON LINE Sample Name The sample name is a text entry box in which you can label the run Type a unique descriptor PeakTrak saves the run info
19. 4 TCP IPv4 and click the Properties button The PC displays the Internet Protocol Version 4 TCP IPv4 Properties window Figure 7 2 M Note If the Internet Protocol Version 4 TCP IPv4 option is not listed click the Install button to add it If assistance is needed press the F1 keyboard button to display Windows Help The following steps change the local area connection settings If the PC must be restored to its original network configuration record the present Internet Protocol TCP IP settings now 5 Select the Use the following IP address option 6 Enter the IP address 192 168 1 200 7 Enter the Subnet Mask 255 255 255 0 8 Click the OK button to close the Internet Protocol Version 4 TCP IPv4 Properties window Click the OK button to close the Local Area Connection properties window 9 Shut down the PC and place the CombiFlash Rf system in Standby CombiFlash Rf User Manual Networking Connect using V VMware Accelerated AMD PCNet Adapter This connection uses the following items 1E Client for Microsoft Networks 8 QoS Packet Scheduler v v IV dB File and Printer Sharing for Microsoft Networks IV 4 Intemet Protocol Version 6 TCP IPv6 a n v v Intemet Link Layer Topology Discovery Mapper I O Driver 4 Link Layer Topology Discovery Responder JOa Transmission Control Protocol Intemet
20. 8 The post run valve cleaning fluids share a common port for channels 2 through 4 Select the 48 inch 120 cm length of black tubing from the accessory package and connect it to the Valve Waste Out bulkhead fitting on the Rf 4x module Route the other end of this tubing to one of the waste bottles The channel 2 waste bottle is shown in Figure 4 8 You can alternatively route it to a container other than those used for channels 2 through 4 however the system will not be able to detect the level and prevent overfilling 4 9 CombiFlash Rf User Manual ae To Waste Bottle 2 Figure 4 8 Valve Waste Out routed to Channel 2 When using higher density solvents such as dichloromethane DCM ensure that waste containers are no more than 3 feet below the supply solvent levels Placing solvent containers above the system and waste containers on the floor may cause the internal check valves to open and allow solvent to flow through the system when in the standby state M Note The Hf 4x back panel ports labeled A and B are not used Section 4 Preparation Optional 4x Module 4 6 Control Connection Operation of the Rf 4x module is controlled by the CombiFlash Rf system via a USB connection Connect the USB cable from the Rf 4x module to USB port on the system back panel 4 7 Position the System After completing the fluid air and control connections the system can be moved to its operating position N CAUTION U
21. 8 Maintenance If the numerical values of the Raw Lamp Energy Figure 8 5 do not change or if the first number remains at zero contact Teledyne Isco s Technical Service department Table 8 1 Suggested Flow Cell Cleaning Solvents Dichloromethane DCM Solvent Description Methanol Miscible with most LC solvents Methanol will force other solvents through the system so they will not interfere with the next sample It breaks down the tertiary structure of silica and allows it to flush through Acetone Common organic solvent with high dissolving properties Acetone absorbs UV light at 254 nm Therefore the Raw Lamp Energy gauge will always read red when acetone is in the flow cell After pumping acetone pump a different solvent through the system to remove the acetone and read the Raw Lamp Energy gauge Effective at solubilizing organic molecules Dimethylsulfoxide DMSO Strong solvent that will clear organic sample Dimethylformamide DMF Strong solvent if other solvents do not produce the desired results aqueous solution Tetrahydrofuran Strong solvent THF Water DMF A 50 25 25 solvent mixture effective in clearing some methanol obstructions Water DMF 30 30 30ml solvent mixture effective in clearing some Acetic Acid obstructions Mildly acidic 0 5M acetic acid in distilled water can be used to clear aqueous solution samples that are not removed by organic solvents Mildly
22. An automatic run extension is a five minute isocratic hold added to the end of the run s maximum B gradient profile During the extension the system continues to pump the maximum B solvent mixture Should the system still detect a peak after an extension the system will add another up toa maximum of three extensions Occasionally compounds might come off the column once the B returns to the minimum value at the end of a run If the Run Length Extension is enabled it will automatically extend the run one time to clear the column and plumbing of any remaining material Extension 1 Extension 3 Minutes Figure 3 9 Automatic Run Extension Examples Figure 3 9 illustrates the possible run extensions The original method was programmed to rise to the maximum B over seven minutes hold for two minutes and return to the minimum B for a final minute At nine minutes a peak was eluting The system extended the run holding the B at the maximum level for another five minutes This also occurred at fourteen and nineteen minutes resulting in the second and third extensions At 24 minutes the B solvent strength returned to zero Before the final minute elapsed more compound was detected causing the system to extend the run for a final five minutes at the minimum B 3 16 Section 3 Preparation Rf 150 Systems Enable rapid equilibration select this option to equilibrate the column at a high flow rate M Not
23. B The gradient plot area also provides a convenient method to edit the gradient curve You can click and drag any of the points to change the shape of the curve or use the buttons at the bottom of the plot area to insert and delete points The following controls appear in the Gradient Plot Area Insert Click this button to enable the gradient point insert mode When this mode is active click the gradient curve to add a single point You can then drag the new point to any desired position Click the insert button once for each gradient point that must be added to the plot area Delete Click this button to enable the gradient point delete mode When this mode is active the system will delete the point nearest the next click on the gradient curve Click the Delete button once for each point that must be deleted from the plot area Run Length Type or select the length of the run Note that this will change the scale of the X axis on the gradient Points that define the gradient will be automatically scaled to fit the new run length The run length can be expressed in minutes or COLUMN VOLUMES on LINE CV You can configure PeakTrak s default Run Length units by changing the CONFIGURATION 6 3 10 settings Solvent A Select the desired solvent from the list The available solvents are those defined by the CONFIGURATION 6 3 10 settings Solvent B Select the desired solvent from the list You can progr
24. Column buttons Click the Stop button to halt the air purge 6 4 10 3 Raw Lamp Energy The right side of the Manual Control window shows a Raw Lamp Energy gauge This provides an indication of the UV light measured by the optical detection system at 254 nm High lamp energy green means that the flow cell easily passes through a sufficient UV light source Lower lamp energy yellow or red means that either the light source is weak or that the flow cell is obstructed Green lamp energy is sufficient to detect peaks up to 2 4 Absorbance Units Yellow lamp energy is partially obstructed and the peak detector performance might be impaired The system can be operated although the absorbance trace might appear noisy Yellow might be a normal indication for systems with flow cell windows stained by compounds Red lamp energy is obstructed to a degree that the system might not reliably detect peaks If you attempt to 6 50 Section 6 User Help Reference operate the system peak collection will be forced to collect all This prevents diverting desired compounds to waste Depending on what you are doing with the Manual Control functions low lamp energy could be a normal indication For example a UV absorbing compound could be present in the flow cell as you are pumping solvent Or the selected solvent absorbs UV light at 254 nm Abnormal indications could be a flow cell blocked by a compound that has precipitated or there
25. Files to Different Column Sizes If you want to scale a method file to a different column size use the FILE gt Save As menu command From the FILES 6 3 6 window select the Scale option then choose the new column size and click the Save button The system will create a new method file with the gradient and run parameters scaled to the new column size This feature is useful when developing methods using small sample volumes After you optimize the method to successfully extract the compounds of interest you can then scale up the method to load and extract larger volumes using larger RediSep column sizes 6 4 4 Editing a Default Method Default Methods allow you to quickly recall standard method settings There are standard methods for each size and type of RediSep column When shipped from the factory the Default Methods were set to the settings recommended by Teledyne Isco The Default Method settings can be tailored to your application To modify the Default Methods 1 Select the TOOLS gt EDIT DEFAULTS 6 2 3 4 menu command 2 If prompted for a password enter the system password The SET PASSWORD 6 3 11 window controls the password protection feature 3 Select the column size and media from the RediSep Column list 4 Change the settings to meet your specific application requirements Refer to the EDITING A METHOD 6 4 2 instructions 5 Click the Save button or File gt Save to replace the default method settings
26. Install Collection Tube Racks Before beginning a run you must load collection racks with tubes onto the system s fraction collector tray Your system was shipped with two collection tube racks The following tube rack sets are available 60 5237 013 Two racks for 13 x 100 mm test tubes 8 mL Total tubes 216 60 5237 061 Two racks for 16 x 100 mm test tubes 14 mL Total tubes 150 Section 2 Preparation Rf 200 and Rf 200i Systems 60 5237 031 Two racks for 16 x 125 mm test tubes 15 5 mL Total tubes 150 60 5237 032 Two racks for 16 x 150 160 mm test tubes 18 mL Total tubes 150 60 5237 033 Two racks for 18 x 150 mm test tubes 25 mL Total tubes 140 60 5237 034 Two racks for 18 x 180 mm test tubes 80 mL Total tubes 140 60 5237 035 Two racks for 25 x 150 mm vials 50 mL Total vials 60 60 5237 040 One rack for twelve 480 mL French square bottles e 60 5394 469 Two racks for 20 mL 28 x 61 mm scintillation vials Total vials 54 60 5394 468 Two racks for 40 mL 28 x 95 mm scintillation vials Total vials 54 To load the racks 1 Insert test tubes vials or bottles into the rack Fig 2 6 Z CAUTION Risk of broken glass or equipment damage Do not load test tubes longer than the length listed on the tube size label Figure 2 6 Loading test tubes 2 13 CombiFlash Rf User Manual 2 While holding the rack with the tube
27. MANAGEMENT 6 2 3 7 6 32 Section 6 User Help Reference HELP gt SERVICE 6 2 4 2 TOOLS gt SET PASSWORD 6 2 3 6 This protects the current password 6 3 12 User Management Use the TOOLS gt USER MANAGEMENT 6 2 3 7 menu command to open a window from which you can add and remove system users When you add a user the system will create a user folder that becomes the data path for that user User management in conjunction with this data path feature conveniently lets you store your working files apart from other system users To add a user click the Add New button and enter the user name To delete a user click the Delete button next to the user s name and confirm the action See also SET DATA PATH 6 3 7 6 4 Examples of PeakTrak actions In this section you will find instructions for typical PeakTrak tasks such as e STARTING A RUN USING DEFAULT SETTINGS 6 4 1 EDITING A METHOD 6 4 2 e ALTERNATIVE WAYS TO CREATE METHOD FILES 6 4 3 e EDITING A DEFAULT METHOD 6 4 4 DEFINING A GRADIENT 6 4 5 REAL TIME GRADIENT EDITING 6 4 6 BYPASSING THE SOLID LOAD CARTRIDGE 6 4 7 MONITORING THE WAVELENGTH RATIO 6 4 8 e VIEWING RUNS 6 4 9 MANUAL CONTROL OF THE COMBIFLASH RF 6 4 10 e SETTING THE VAPOR LIMIT 6 4 11 6 4 1 Starting a run using default settings After the CombiFlash Rf system has been set up and automatically primed See AUTO PRIME VALVE WASH 6 3 8 6 33 CombiF
28. Network Type Select the Static IP option This option will require an IP Address Netmask and Gateway provided by your network administrator P Address Enter the static IP Address for the system Netmask Enter the Netmask for the system Gateway Enter the Gateway for the system Network Printing The system can print to a network printer and supports JetDirect and line printer LPR queues Consult with your network administrator to determine The IP address of selected printer e Ifthe printer uses an LPR print queue you must also find out the queue name e Ifthe printer uses a JetDirect print queue also ask for the port number The printer type either Postscript or PCL When this information is known you can proceed with configuring system for network printing 6 30 Section 6 User Help Reference Follow the on screen instructions for entering the address queue and printer type information After successfully printing a test page the network printer will be available for printing using the FILE gt PRINT 6 2 1 12 menu command and the AUTOMATICALLY PRINT REPORT AT END OF RUN feature The Network printing controls may also be configured to print to a local USB printer connected the system s back panel USB port The printer must support postscript printing To configure the local printer select Postscript for the Printer Type Then type the following connection URL usb dev us
29. Rf system 1 6 Safety Before installing operating or maintaining this equipment it is imperative that all hazards and preventive measures are fully understood While specific hazards may vary according to location and application take heed in the following general warnings A WARNING Avoid hazardous practices If you use this instrument in any way not specified in this manual the protection provided by the instrument may be impaired A WARNING Liquids associated with this instrument may be classified as carcinogenic biohazard flammable or radioactive Should these liquids be used it is highly recommended that this application be accomplished in an isolated environment designed for these types of materials in accordance with federal state and local regulatory laws and in compliance with your company s chemical hygiene plan in the event of a spill A WARNING If you are using flammable solvents or chemicals with this system vapor concentration levels may exceed the maximum exposure levels as recommended by OSHA Guide 1910 1000 To reduce those levels to a safe exposure Teledyne Isco recommends that you place the system in a laboratory hood designed for the purpose of ventilation This hood should be constructed and operated in accordance with federal state and local regulations In the event of a solvent or chemical spill your organization should have a plan to deal with these mishaps In all cases use good labora
30. Risk of broken glass or equipment damage Do not load test tubes longer than the length listed on the rack label 2 While holding the rack with the rack label visible Figure 3 7 insert the racks into the system Slide the rack in until you feel it drop into its seated position Rack Label Figure 3 7 Inserting the collection tube rack 3 11 CombiFlash Rf User Manual CAUTION An incorrectly installed rack will cause the rack to be misaligned under the fraction collector arm Misaligned racks might cause fractions to miss the tube opening or deposit in the wrong tube Always ensure the tube size label is visible that is facing outward and the rack is pushed in until it is seated 3 9 Turn on Power The system s power switch is located just below the touch screen panel 1 Turn the switch to the ON position The system will begin its startup routine which includes self diagnostics 2 Monitor the movement of the fraction collector arm The arm should move to the left rear corner of the fraction collector area NCAUTION You should not hear any grinding noises during the homing sequence If grinding is heard there is a problem with the homing of the fraction collector arm Contact the factory for assistance 3 The system is ready for operation when the PeakTrak screen is displayed 3 10 Configure the System Before operating the system it should be configured for the desired operation
31. Section 2 Preparation Rf 200 and Rf 200i Systems of level bench space Ensure that the module has at least 3 cm 1 25 of air space behind it for ventilation Instructions for installing the 4x module can be found in Section 4 of this guide Refer to Table 1 1 for environmental conditions and power requirements Z WARNING The system is heavy Use a two person lift to prevent injury Before making any connections to the Combiflash Rf system place it on the bench or in the fume hood where it will be operated Temporarily position the system so you can access the back panels to complete the connections 2 3 Connect Power Ensure that the On Standby switch below the touch screen panel is in the Standby position Then use the supplied IEC power cord to connect the system to mains power A WARNING Mains power must meet the voltage frequency and amperage requirements listed on the serial number label A WARNING As long as the AC mains power cord is connected power is inside the unit The mains power cord is the disconnect device Position the system so that the power cord can be unplugged or use a power strip where the plug can quickly be removed from the outlet in the event of an emergency 2 4 Connect Solvent Lines A WARNING Risk of fire ignited by electrostatic discharge Never substitute the black tubing on CombiFlash systems The black tubing P N 023 0503 06 is conductive This tubing is required to dissip
32. Start a Default Method LL 5 4 During the Run KK KK KK RR RR RR KK eee 5 4 1 Run Control Buttons 5 4 2 Bypassing the Solid Load Cartridge 5 4 3 About Solvent Level Detection 6 User Help Reference 6 1 PeakTrak Overview 0 KK RR KK KK 6 1 1 PeakTrak Window Elements 6 1 2 Method File 0 0 0 0 KK KK eee 6 L9 Run Bie i 3an KA nade bee CEES SR ME kk 6 1 4 How to get PeakTrak help 6 2 PeakTrak Menu Options 0005 22 T Elle Lore tact aden ous A RE d 6 2 2 Method Editor 0 0 20 6 2 3 TOONS 5 gag pues ein co lS Ale Whee CORE epos 6 24 Help uote alee a n eens cues de as 6 3 PeakTrak Windows 00000 RR KK K 6 3 1 Main Window 0 0000 RR RR KK 6 3 2 Method Editor 0 002000 6 3 8 Gradient Optimizer 05 6 3 4 Column Data 0 KK KK 6 3 5 Batch eee RR ve ae RA 6 3 6 Eleg ace ced rese oe Pals Rd S res 6 3 7 Set Data Path RR 6 3 8 Auto Prime Valve Wash 6 3 9 Manual Control KK 6 3 10 Configuration llle 6 3 11 Set Password 0 eee eee eee 6 3 12 User Management sse 6 4 Examples of PeakTrak actions 6 4 1 Starting a run using default settings 6 4 2 Editing amethod 005 6 4 3 Alternative Ways to Create Met
33. The Cartridge Bypass button may be used during a run to relieve backpressure caused by impurities precipitating in the solid load cartridge Use this feature with care to avoid leaving desired compound on the cartridge Read more about this feature in BYPASSING THE SOLID LOAD CARTRIDGE 6 4 7 If the external detector option is enabled on Rf 200 systems the plot area will include a secondary left Y axis to display the analog input voltage trace Likewise if the evaporative light scattering detector ELSD is enabled on an Rf 200i system a secondary left Y axis is included The gradient plot area also provides a convenient method to edit the gradient curve You can click and drag any of the points to change the shape of the curve and use the buttons at the bottom of the plot area to insert and delete points For complete instructions on defining gradient curves see DEFINING A GRADIENT 6 4 5 To zoom in on part of the plot area click the Zoom In button to enable the zoom action and then drag across the area of interest Click the Zoom Out button to revert back to the normal display 6 3 1 2 Run Settings RediSep Rf200 Rf200i and Rf4X systems The system reports the detected RediSep column media and size The column should match the one that will be used for the run You may override the detected column size by selecting a different size or media See also REDISEP RF COLUMN SELECTION GUIDE oN LINE and ABOUT REDISEP RF COLUM
34. The file name will be the same as the Run file except with a TXT file name extension When selecting this command from a remotely connected computer via a web browser PeakTrak displays a file download window so you can select a location and enter a file name You can open edit and print TXT files with text editing or word processing software 6 5 CombiFlash Rf User Manual 6 2 1 9 Import Method Choose this command to load a Method file from an external source see EXPORT METHOD 6 2 1 10 onto the system s internal storage From the touch screen this command opens the FILES 6 3 6 window so you can locate and select the file on a USB Flash storage device When selecting this command from a remotely connected computer PeakTrak displays a window so you can select the method file After selecting the file click the Open button to save the method file on the system s internal hard drive 6 2 1 10 Export Method Choose this command to save a Method file in a location other than the system s internal hard drive This can be done to archive method files or to transfer the method to another system using the IMPORT METHOD 6 2 1 9 menu command From the touch screen this command opens the FILES 6 3 6 window so you can save the file on a USB Flash storage device When selecting this command from a remote computer PeakTrak displays a window from which you can save the method file 6 2 1 11 Export SMM XL Method Choose
35. Zone Select your time zone from the drop down list box 3 10 3 Set Date Time Click this button to open a window from which you can set the system date and time To prevent unauthorized time changes this feature is password protected The system is shipped with the password set to combiflash Use the Tools gt Set Password menu command to change this password for greater security 3 10 4 Vapor Limit Optional The system has internal vapor The sensor detects vapors present in the immediate atmosphere When the limit is exceeded the CombiF lash Rf will shut down to avoid a hazardous condition Teledyne Isco recommends using the default setting of 25 This value represents an organic vapor level slightly above the ambient vapor level of a well ventilated laboratory A setting below 25 could cause random alarms without significant vapors present If PeakTrak displays a Vapor Limit 3 13 CombiFlash Rf User Manual alarm while the Vapor Limit is set at 25 perform the following checks on your laboratory and the instrument e Ensure that no open containers or spills of organic solvent are in close proximity to the system e Ensure that the system is located in a well ventilated area e Ensure that there is no visible solvent leakage from the system If PeakTrak continues to display the Vapor Limit alarm after you have made these checks and corrected any problems found it is likely that organic vapors are present in t
36. and is collected An automatic run extension is a five minute isocratic hold added to the end of the run s maximum 4B gradient profile During the extension the system continues to pump the maximum B solvent mixture Should the system still detect a peak after an extension the system will add another up to a maximum of three extensions Occasionally compounds might come off the column once the B returns to the minimum value at the end of a run If the Run Length Extension is enabled it will automatically extend the run one time to clear the column and plumbing of any remaining material Original method Extension 2 Final Extension Extension 1 Extension 3 15 20 25 Minutes Figure 2 9 Automatic Run Extension Examples Figure 2 9 illustrates the possible run extensions The original method was programmed to rise to the maximum B over seven minutes hold for two minutes and return to the 2 19 CombiFlash Rf User Manual minimum B for a final minute At nine minutes a peak was eluting The system extended the run holding the B at the maximum level for another five minutes This also occurred at fourteen and nineteen minutes resulting in the second and third extensions At 24 minutes the B solvent strength returned to zero Before the final minute elapsed more compound was detected causing the system to extend the run for a final five minutes at the minimum B Enabl
37. below the level of the Rf can contribute to decreased flow due to the high vapor pressure of DCM This problem becomes more pronounced as the ambient temperature increases CombiFlash Rf User Manual 3 8 Install Collection Tube Racks Before beginning a run you must load collection racks with tubes onto the system s fraction collector tray Your system was shipped with two collection tube racks The following tube rack sets are available 60 5237 013 Two racks for 13 x 100 mm test tubes 8 mL Total tubes 216 60 5237 061 Two racks for 16 x 100 mm test tubes 14 mL Total tubes 150 60 5237 031 Two racks for 16 x 125 mm test tubes 15 5 mL Total tubes 150 60 5237 032 Two racks for 16 x 150 160 mm test tubes 18 mL Total tubes 150 60 5237 033 Two racks for 18 x 150 mm test tubes 25 mL Total tubes 140 60 5237 034 Two racks for 18 x 180 mm test tubes 30 mL Total tubes 140 60 5237 035 Two racks for 25 x 150 mm vials 50 mL Total vials 60 60 5237 040 One rack for twelve 480 mL French square bottles 60 5394 469 Two racks for 20 mL 28 x 61 mm scintillation vials Total vials 54 60 5394 468 Two racks for 40 mL 28 x 95 mm scintillation vials Total vials 54 3 10 Section 3 Preparation Rf 150 Systems To load the racks 1 Insert test tubes vials or bottles into the rack Fig 3 6 Figure 3 6 Loading test tubes A CAUTION
38. by Tele dyne Isco 7 2 Section 7 Remote Interfaces To access the system simply open an Internet Browser and enter the IP address as http __ _ where the blanks are replaced by the selected address The browser window will load PeakTrak after you enter the address 7 1 2 Network Printing The CombiFlash Rf system can print to a network printer and supports both JetDirect and line printer LPR queues Consult with your network administrator to determine the IP address of selected printer If the printer uses an LPR print queue you must also find out the queue name If the printer uses a JetDirect print queue also ask for the port number When this information is known you can proceed with configuring system for network printing 1 Select the Tools Configuration menu command 2 Select the Network Configuration tab 3 Follow the on screen instructions for entering the address and queue information 4 Click OK to save the settings The Combiflash Rf system will send a test page to the printer After successfully printing a test page the network printer will be available for printing using the File gt Print menu command or through the Automatically Print Report at End of Run option on the Tools gt Configuration User Preferences tab 7 1 3 Network File Save Configuration By default this option is disabled and post run reports are automatically saved internally on the CombiFlash Rf system You can recal
39. complete the run the run continues If not sufficient the system displays the alert message again Ignore This option is available when the estimated usage rates indicates that the solvent levels are insufficient Select Ignore to clear the message and continue the run The system then ignores the estimated solvent usage rate for the remainder of the run but continues to measure the solvent level Should the level become insufficient the system will display another alert message Disable Select this option to turn off solvent level measurements and estimated solvent usage rates for the remainder of the run 5 14 Section 5 Operation Cancel This option appears before the run is started Select this option to cancel the run and return to the main window which will display the current method 5 15 CombiFlash Rf User Manual CombiFlash Rf User Manual Section 6 User Help Reference The CombiFlash Rf system includes User Help topics that are available from the PeakTrak touchscreen or remote browser window This section contains many of these topics that describe the PeakTrak interface and its use On line Software Help benefits from linked text to navigate through topics of interest a feature not available in printed manuals For your convenience linked text is displayed in SMALL CAPITALIZED TEXT followed by the section number in brackets Some topics are only available in the on line help
40. containers are filled then 1 Select the Tools gt Auto prime menu command 2 Click the Play button to start priming the system The system pumps 100 Solvent B through the system and out the Waste port and then repeats with 100 Solvent A This B A order leaves the internal lines filled with solvent appropriate for the beginning of a purification run When priming the first time inspect the solvent and waste connections to the system If any leaks are observed click the Cancel button to stop the Auto Prime Correct the leak by tightening the fitting an additional 1 4 turn and then restart the Auto Prime from step 1 To advance to the next Auto Prime step you can click the Fast Forward button The Auto Prime window closes when finished After Auto Priming the system is ready for operation Section 5 Use the Auto Prime feature for quick solvent changes by pumping a fixed amount of solvent through the waste fluid path at a high flow rate For more advanced priming functions such as varying the flow rates fluid paths etc use the Tools gt Manual Control option 3 18 Section 3 Preparation Rf 150 Systems 3 12 System Verification It is recommended that the system operation be verified To do so use a pre packaged CombiFlash Rf test kit Part number 60 5237 050 Test kit containing five of the 5 gram size solid load cartridges pre filled with 0 5 grams of test sample NPHE and five 4 gram RediSep Rf sil
41. de las instrucciones del manual que acompa an a este producto Esta se al alerta sobre la presencia de alto voltaje en el interior del producto Punto del machacamiento Sus dedos o manos seriusly A ser n da ados si usted los coloca entre las piezas m viles cerca de estos s mbolos 1 7 For Additional Information Technical assistance for the CombiFlash Rf system can be obtained from Teledyne Isco 4700 Superior St Lincoln NE 68504 Phone 800 775 2965 or 402 464 0231 Fax 402 465 3001 E mail IscoService teledyne com 1 22 CombiFlash Rf User Manual Section 2 Preparation Rf 200 and Rf 200i Systems This section provides instructions for unpacking and installing the CombiFlash Rf 200 and Rf 200i systems To prepare a system for operation sequentially follow all instructions in sections 2 1 through 2 13 M Note Section 2 15 contains an Installation Qualification checklist If required sign off the checklist entries as you successfully complete the following sections 2 1 Unpacking the Unit The CombiFlash Rf 200 or 200i system is shipped in a single carton The optional 4x expansion module is shipped in a second carton Carefully unpack the shipment and inspect the contents NWARNING The system is heavy Use a two person lift to prevent injury NCAUTION Do not lift the system by the fraction collector arm Use the lifting handles located on the instrument s
42. is a film built up on the flow cell If you suspect the low lamp energy is an abnormal indication perform the steps in the QUICK CLEANING WHEN RECOMMENDED ON LINE topic 6 4 11 Setting the Vapor Limit Rf200 Rf200i and Rf4X The system has an internal vapor sensor that detects vapors present in the immediate atmosphere When the limit is exceeded the CombiFlash Rf will shut down to avoid a hazardous condition Teledyne Isco recommends using the default setting of 25 This value represents an organic vapor level slightly above the ambient vapor level of a well ventilated laboratory A setting below 25 could cause random alarms without significant vapors present If PeakTrak displays a Vapor Limit alarm while the Vapor Limit is set at 25 perform the following checks on your laboratory and the instrument Ensure that no open containers or spills of organic solvent are in close proximity to the system Ensure that the system is located in a well ventilated area Ensure that there is no visible solvent leakage from the system If PeakTrak continues to display the Vapor Limit alarm after you have made these checks and corrected any problems found it is likely that organic vapors are present in the ambient environment of your laboratory In this case increase the setting of the Vapor Limit Settings greater than 25 are appropriate for laboratory environments with a somewhat elevated background solvent vapor concentration rep
43. laboratory has more than one Combiflash Rf system Time Zone Select your time zone from the drop down list box 2 12 3 Set Date Time Click this button to open a window from which you can set the system date and time To prevent unauthorized time changes this feature is password protected The system is shipped with the password Section 2 Preparation Rf 200 and Rf 200i Systems set to combiflash Use the Tools gt Set Password menu command to change this password for greater security 2 12 4 Vapor Limit The system has an internal vapor sensor that detects vapors present in the immediate atmosphere When the limit is exceeded the Combiflash Rf will shut down to avoid a hazardous condition Teledyne Isco recommends using the default setting of 25 This value represents an organic vapor level slightly above the ambient vapor level of a well ventilated laboratory A setting below 25 could cause random alarms without significant vapors present If PeakTrak displays a Vapor Limit alarm while the Vapor Limit is set at 25 perform the following checks on your laboratory and the instrument e Ensure that no open containers or spills of organic solvent are in close proximity to the system e Ensure that the system is located in a well ventilated area e Ensure that there is no visible solvent leakage from the system If PeakTrak continues to display the Vapor Limit alarm after you have made these checks and corrected any probl
44. selection valve and internal components M Note When using higher density solvents such as dichloromethane DCM place the solvent container level with or above the CombiFlash Rf system Placing solvent containers below the level of the system can contribute to decreased flow due to the high vapor pressure of DCM This problem becomes more pronounced as the ambient temperature increases 3 5 CombiFlash Rf User Manual 3 5 Connect Waste Lines A WARNING Risk of fire or equipment damage Failure to connect Waste Port tubing may allow organic solvents to pool in unsafe areas possibly creating dangerous levels of flammable vapors Z WARNING Risk of fire ignited by electrostatic discharges Never substitute the black tubing on CombiFlash systems The black tubing P N 023 0503 06 is conductive This tubing is required to dissipate static electricity NCAUTION Elevated flammable vapor levels are possible Ensure that the waste container is adequately ventilated preferably by placing it in a fume hood CAUTION Do not overfill the waste collection containers Regularly monitor and empty the bottles before they overfill The system has one waste outlet on the back panel Diverter Valve waste Accessory package P N 60 5239 009 includes a Waste Tube Assembly P N 60 5234 187 To connect the Waste tube 1 Connect the cut end of this assembly to the Diverter Valve Waste port on the system a locate the blac
45. solvent connections 1 Locate the ABCD label in accessory package 69 5234 008 Affix this label to the system s left side panel just above the solvent ports Figure 4 1 A B C D Figure 4 1 Labeled ports on CombiFlash Rf system side panel 2 Remove the jumper tubing from ports A B C and D 3 Locate the solvent lines on the side of the Rf 4x module These lines are labeled A through D Connect the solvent lines to their respective ports on the CombiFlash Rf system 4 3 CombiFlash Rf User Manual 4 5 Connect Waste Lines A WARNING Risk of fire or equipment damage Failure to connect Waste Port tubing may allow organic solvents to pool in unsafe areas possibly creating dangerous levels of flammable vapors A WARNING Risk of fire ignited by electrostatic discharges Never substitute the black tubing on CombiFlash systems The black tubing P N 023 0503 06 is conductive This tubing is required to dissipate static electricity N CAUTION Elevated flammable vapor levels are possible Ensure that the waste container is adequately ventilated preferably by placing it in a fume hood The Rf 4x module has six liquid waste ports on the back panel Figure 4 4 one inlet to receive diverter valve waste from the CombiFlash Rf system labeled Column Waste From RF four waste outlets one for each column channel labeled Column Waste 1 2 3 and 4 one Valve Waste outlet shared by
46. system uses air or user supplied gas to measure the hydrostatic pressure of the waste above the level sense line end The system will suspend operation when this level is too high This port is not present on the Rf 150 systems Diverter Valve Waste The diverter valve on the fraction collector arm sends fluid to this waste port Inject Valve Waste The injection valve is cleaned after each run The system directs fluids from this automatic cleaning routine to this port 25 Lower Drain Tube This tube carries away liquids spilled onto the rack area 26 Cooling Fan Rf 200i only Cools the internal ELSD components Section 1 Introduction 27 Exhaust Port Rf 200i only Vents the carrier gas and vaporized solvents Refer to Fig 1 7 for other back panel features Figure 1 8 CombiFlash Rf 200i ELSD Features Back 28 Nitrogen Inlet Rf 200i only Connects to the carrier gas supply 60 to 70 psig 29 External Detector Rf 200i only Reserved for future use 30 P Trap Drain Rf 200i only During operation condensate from the Thermo split process drains from the system An internal P trap prevents aerosol particles from escaping through this drain 31 Pump Drain Rf 200i only An internal drip tray is located below the splitter pump for the ELSD Should the pump or its fittings leak liquids will drain from this port to protect internal components CAUTION Discontin
47. the system operation be verified To do so use a pre packaged CombiFlash Rf test kit e Part number 60 5237 050 Test kit containing five of the 5 gram size solid load cartridges pre filled with 0 5 grams of test sample NPHE and five 4 gram RediSep Rf silica gel columns e Part number 60 5237 051 Test kit containing four of the 25 gram size solid load cartridges pre filled with 0 5 grams of test sample NPHE and four 4 gram RediSep Rf silica gel columns 2 15 Installation Qualification Checklist Table 2 1 may be completed to verify and document the installation procedures contained in Section 2 of this guide 2 22 Section 2 Preparation Rf 200 and Rf 200i Systems Table 2 1 Installation Qualification Checklist Installer Operator Description Initials Initials UNPACKING THE UNIT INSTRUMENT LOCATION CONNECT POWER CONNECT SOLVENT LINES CONNECT WASTE LINES 2 6 EXTERNAL GAS Optional 2 7 CONNECT AND ROUTE DRAIN LINES 2 8 POSITION THE SYSTEM 2 9 INSTALL COLLECTION TUBE RACKS 2 10 EXTERNAL DETECTOR OPTIONAL 2 11 TURN ON POWER 2 12 CONFIGURE THE SYSTEM 2 13 PRIME THE SOLVENT LINES 2 14 SYSTEM VERIFICATION Certification of Section 2 Completion Installer Name print Installer Signature Date Operator Name print Operator Signature Date Comments 2 23 CombiFlash Rf User Ma
48. this command to export a method file for use on a CombiFlash Companion system equipped with the Solvent Management Module SMM or a CombiFlash Companion XL system From the touch screen this command opens the FILES 6 3 6 window so you can save the file on a USB Flash storage device When selecting this command from a remote computer PeakTrak displays a window from which you can save the method file 6 2 1 12 Print From the touch screen this command prints the completed run on the network printer see NETWORK PRINTING on the NETWORK CONFIGURATION 6 3 10 2 window When selecting this command from a remotely connected computer PeakTrak displays the Printer window From this window you can select and configure the printer of your choice After configuring the settings click the PRINT button to print the completed run 6 6 Section 6 User Help Reference 6 2 1 13 Logout Choose this command to logout of the system After logging out the system displays a login screen and waits for the next user to log in 6 2 2 Method Editor Selecting the Method Editor menu command will open the METHOD EDITOR 6 3 2 window Use this window to view and modify the advanced method file settings 6 2 3 Tools PeakTrak s Tools menu has the following options 6 2 3 1 Auto Prime Valve Wash Choose this command to automatically prime the solvent pumps On CombiF lash Rf 200 and Rf 200i systems this feature will also wash the injection valv
49. waste collection container Drain tubing is not required on the Pump Drain port NCAUTION Discontinue use of the CombiF ash Rf 200i if liquid is present at the Pump Drain Contact Teledyne Isco technical service for assistance with correcting the leak CombiFlash Rf User Manual 2 8 Position the System After completing the various connections the system can be moved to its operating position Turn the system so that the operator can access all of the front view features and controls Figure 1 5 Use care not to damage the connections tubing and cables while moving the system CAUTION Ensure that the CombiFlash Rf has at least 3 cm 1 25 of air space behind it for ventilation Position the solvent and waste containers as necessary When using higher density solvents such as dichloromethane DCM place the solvent container level with or above the CombiFlash Rf system Placing solvent containers below the level of the Rf can contribute to decreased flow due to the high vapor pressure of DCM This problem becomes more pronounced as the ambient temperature increases When using higher density solvents such as dichloromethane DCM ensure that the waste container is no more than 3 feet below the supply solvent levels Placing solvent containers above the system and waste containers on the floor may cause the internal check valves to open and allow solvent to flow through the system when in the standby state 2 9
50. 1000 4 3 2006 Radiated RF Immunity 80 mHz to 2 7gHz 80 AM at 1 kHz 1A1 2007 A2 2010 Level 2 3V m A EN61000 4 4 2004 Electrical Fast Transient EFT on Level 2 1kV on AC lines A 1A1 2010 Mains and I O EN61000 4 5 2005 Surge on AC Lines Level 2 1kV Line Common Mode A Level 2 0 5kV Differential Mode EN61000 4 6 2008 Conducted RF Immunity on Mains and 150 kHz to 80 mHz A vo Level 1 1V rms 80 Modulated EN61000 4 11 2004 Voltage Dips 0 during cycle and full cycle A 70 at 25 cycles CISPR11 RF Emissions Radiated below 1GHz Group 1 Class A Industrial Scientific and PASS EN 55011 2009 and Conducted AC Mains Medical Equipment A1 2010 EN61000 3 2 2005 AC Harmonics Flicker PASS A1 2008 A2 2009 EN61000 3 3 2008 We the undersigned hereby declare that the design of the equipment specified above conforms to the above Directive s and Standards as of March 13 2013 USA Representative Lb Rte Vikas V Padhye Ph D Vice President and Product General Manager 4700 Superior Street Lincoln Nebraska 68504 402 464 0231 402 464 0318 60 5232 509 Rev B TELEDYNE ISCO j Everywhereyoulook iW TELEDYNE ISCO A Teledyne Technologies Company 4700 SUPERIOR ST PHONE 402 464 0231 P O BOX 82531 FAX 402 464 0318 LINCOLN NE 68504 2531 Effective January 5 2012 Subject Teledyne Isco Products RE Bovine Spongiform
51. CONFORMITY ISMEA Application of Council Directive Manufacturer s Name Manufacturer s Address Equipment Type Environment Trade Name Model No Year of Issue Standards to which Conformity is Declared 2004 108 EC The EMC Directive 2006 95 EC The Low Voltage Directive 2002 96 EC The WEEE Directive Teledyne Isco Inc 4700 Superior Lincoln Nebraska 68504 USA P O Box 82531 Lincoln NE 68501 Phone 1 402 464 0231 Facsimile 1 402 465 3799 Laboratory Equipment for Light Industrial Commercial Environments Combiflash RF4X 2008 EN 61010 1 2 Edition Safety Requirements for Electrical Equipment for Measurement Control and Laboratory Use EN 61326 2006 EMC Requirements for Electrical Equipment for Measurement Control and Laboratory Use Standard Description Severity Applied Performance Criteria EN61000 4 2 Electrostatic Discharge Level 2 4kV contact discharge A Level 3 8kV air discharge EN61000 4 3 Radiated RF Immunity 80 Mhz to 2 7Ghz 80 AM at 1kHz A Level 2 3V m EN61000 4 4 Electrical Fast Transient Level 2 1kV on AC lines A EN61000 4 5 Surge on AC Lines Level 2 2kV common mode Level 2 1KV differential mode EN61000 4 6 Conducted RF on AC lines 150 kHz to 80 MHz A Level 1 1V rms 80 modulated EN61000 4 11 Voltage Dips 1 10 25 cycles 100 40 70 A Voltage Interruption 250 cycles 100 drop c CISPR11 RF Emissions G
52. CombiFlash Rf User Manual For indoor use only COPYRIGHT 2013 by Teledyne Isco 4700 Superior St Lincoln Nebraska U S A 68504 Phone 402 464 0231 Toll Free 800 228 4373 FAX 402 465 3022 Revision A August 13 2013 Part 60 5233 584 CombiFlash Rf User Manual CombiFlash Rf User Manual Table of Contents 1 Introduction 1 1 Documentation Overview 0 000 eee 1 1 1 2 Product Overview 0 0 0 KK KK KK RR KK KK KS 1 1 1 3 Operating Overview 0 0 0 1 6 1 3 1 Multiple Control Possibilities 1 6 1 3 2 File Storage cs wg vere 1 6 1 4 Specifications kK KK KK KK KK KK KK KK KK KI 1 7 1 5 Controls Indicators and Features 1 13 TG Safety aa le Con du e ice ds 1 19 1 6 1 Hazard Severity Levels 1 20 1 6 2 Hazard Symbols 00000 1 20 1 7 For Additional Information 1 22 2 Preparation Rf 200 and Rf 200i Systems 2 1 Unpacking the Unit 0 00085 2 1 2 2 Instrument Location KK KK KK KK KK eee 2 2 2 8 Connect Power 0 eee KK KK 2 3 2 4 Connect Solvent Lines 0 000 cee eee 2 3 2 5 Connect Waste Lines eese 2 6 2 6 External Gas 0 eee eee 2 8 2 6 1 CombiFlash Rf 200 and Rf 200i 2 8 2 6 2 CombiFlash Rf 200i Only 2 9 2 7 Connect and Route Drain Lines 2 9 2 8 P
53. Data Path The top left corner of the window displays the path current folder As you browse through the files the path will update as you go New Click this button to add and name a subfolder within the currently selected folder Delete Click this button to delete the selected folder Folder selection box This box lists the available folders and selection buttons e OK Click this button to save your selection as the data path and close the window Cancel Click the Cancel button to close the window without changing the data path 6 3 8 Auto Prime Valve Wash The Auto Prime feature is a convenient way to prime the pumps with solvent This should be done whenever new solvents are connected to the back panel ports or to change between the available solvents An Auto Prime pumps 40 mL 6 25 CombiFlash Rf User Manual of the selected Solvent B drawing it in through the back panel port through the pump and out the waste port On Rf 200 Rf 200i and Rf 4x systems the injection valve is included in the fluid path to wash the valve The Auto Prime then does the same for the selected Solvent A By priming Solvent A last the system is ready for the starting mobile phase of the next run The Auto Prime window is opened by selecting the TOOLS gt AUTO PRIME VALVE WASH 6 2 3 1 menu command The window contains the following controls Progress bar This bar displays how much of the solvent ha
54. Drain Tubing Process Valves Vinyl with FEP liner PEEK PTFE perfluoroelastomer Sample Loading Cap 316 stainless steel Sample Loading Cap Seal PTFE Elgiloy Injection Valve PTFE PCTFE Flowcell 303 SST Type ES Quartz SIMRIZ SZ485 Pump 316 stainless steel PTFE Elgiloy a Elgiloy is a registered trademark of Combined Metals of Chicago L L C Section 1 Introduction 1 5 Controls Indicators and Features Figures 1 5 through 1 7 illustrate the controls on the CombiFlash Rf Figure 1 5 CombiFlash Rf Features Front 1 Touch Panel LCD display Large 10 5 inch display for system monitoring and control 2 USB Port Convenient front panel port that accepts USB Flash memory drive A Flash drive may be inserted into this port for transferring files importing and exporting methods and performing system software updates 3 Adjustable Column Mount The injection valve assembly and upper column mount slide along this mount so the system can accept a variety of column sizes 4 On Standby Switch 1 On 0 standby 5 Fraction Collector Arm and Drop Former The arm and drop former move to deposit liquid in the collection tubes 6 Racks and Collection Tubes Racks hold the fraction collection tubes for all Rf system Racks include an RFID 1 13 CombiFlash Rf User Manual tag which the system uses to read the rack type and collection tube size
55. Encephalopathy BSE Transmissible Spongiform Encephalopathy TSE Teledyne Isco products are not manufactured using materials that are derived from animal sources Therefore we can state that Bovine Spongiform Encephalopathy BSE Transmissible Spongiform Encephalopathy TSE should not be a concern If you have any questions or require further information please contact us or visit our website at www isco com Sincerely Ut Vikas Padhye Ph D Vice President and General Manager 4700 Superior Street Lincoln Nebraska 68504 Phone 402 464 0231 FAX 402 464 0318 This information is considered accurate and reliable as of the date appearing above and is presented in good faith Because use conditions and applicable laws may differ from one location to another and may change with time recipient is responsible for determining whether the information in this document is appropriate for recipient s use Since Teledyne Isco has no control over how this information may be ultimately used all liability is expressly disclaimed and Teledyne Isco assumes no obligation or liability thereof Therefore no warranty expressed or implied is given nor is freedom from any patent owned by Teledyne Isco or others to be inferred FmtASASMALARNEHRAS Name and amount of Hazardous Substances or Elements in the product BBTA BIRER JR BES BR Hazardous Substances or Elements Component Name E Ld TOME ZBRKA SR
56. KE Pb Hg Cd Cr VD PBB PBDE ARER LCD Display o o o o o o none on RF4X SER x O O O e O Circuit boards Beek O e e e X O Wiring Pas Bet O O O O x O Internal Cables ERR O O O O x O Line Cord SHON X O O O X O Stepper Motor TAN O o X o Oo O Deuterium lamp Rik O e e e x O Valve Body J m h B SUB SWART FAR BB Name and amount of Hazardous Substances or Elements in the product o RAT BT EYI RT ZIN II ARMA PHS RSET MEMENRELRAT O Represent the concentration of the hazardous substance in this component s any homogeneous pieces is lower than the ST standard limitation X BINAB RET EI E 212022 N AR HRM rh ES HERIN ST HMERENHREER EAL IZE CARE ARS Sc ESTE EIT X HERRARTE A X Represent the concentration of the hazardous substance in this component s at least one homogeneous piece is higher than the ST standard limitation Manufacturer may give technical reasons to the X marks FRE FIR 4J0 WASE The Environmentally Friendly Use Period EFUP was determined through experience E EL REHAB TE RNS rh BU PECES E77 45 207 1858200755 BEJE E RHSR BD A A B ZA 5 The date of Manufacture is in code within the serial number The first three numbers are the year of manufacture 207 is year 2007 followed by a letter for the month A is January B is February and so on CombiFlash Hazmat Table 60 5233 406 Rev B Teledyne Isco O
57. N DETECTION OoN LINEI Sample Name The sample name is a text entry box in which you can label the run Type a unique descriptor PeakTrak saves the run information under this name Therefore PeakTrak cannot accept any characters that are reserved by the operating system V etc If you do not type a Sample Name PeakTrak will generate a date time stamp for the name when you click the Play button to begin the run CombiFlash Rf User Manual Flow Rate Type or select the desired flow rate for the run When using the Isco default column methods the system sets the flow rate to the optimum flow rate of the column Detector Rf 200 Rf 200i and Rf 4x systems Reports the current peak detection mode To change the peak detection settings open the METHOD EDITOR 6 3 2 window Solvent A Select the desired solvent from the list The solvents in the list are those defined by the CONFIGURATION 6 3 10 window For assistance in selecting a solvent see the SOLVENT SELECTION ON LINE table Solvent B Select the desired solvent from the list 6 3 1 3 Run Control Buttons From left to right the control buttons are Play The Play button starts or resumes the run After a run has started this button is replaced by the Pause button Pause The Pause button holds the B at the current value while the system continues to operate sometimes called an isocratic hold Note that pausing the run extend
58. ON LINE SOLVENT SELECTION ON LINE EDITING A METHOD 6 4 2 EDITING A DEFAULT METHOD 6 4 4 BYPASSING THE SOLID LOAD CARTRIDGE 6 4 7 6 4 2 Editing a method Editing a method allows you to tailor the operation of the CombiFlash Rf system to best separate or purify the compounds of interest PeakTrak has two types of method settings basic and advanced 6 4 2 1 Basic method settings The basic settings allow you to control the B gradient mix flow rate solvents and run length These settings are part of the MAIN WINDOW 6 3 1 These settings are the most frequently changed settings when developing or improving purification methods The B gradient mix and run length are shown on the plot area Refer to the DEFINING A GRADIENT 6 4 5 topic for more details The other settings are Flow Rate When using the Isco default column methods the flow rate is automatically set to the optimum flow rate of the column If using a non Isco column refer to the manufacturer s column documentation to determine the best flow rate for the column Note that if the column is not detected by the system the operating pressure is limited to 50 psi Solvent A and B Select the desired solvents from the lists The solvents are those defined by the CONFIGURATION 6 3 10 window 6 4 2 2 Advanced method settings The METHOD EDITOR 6 3 2 window gives you access to advanced settings such as equilibration volume initial waste air p
59. Protocol The default wide area network protocol that elevi communication General You can get IP settings assigned automatically if your network supports this capability Otherwise you need to ask your network administrator for the appropriate IP settings Obtain an IP address automatically Use the following IP address TP address Subnet mask Default gateway Obtain DNS server address automatically Use the following DNS server addresses Preferred DNS server Alternate DNS server Figure 7 2 Internet Protocol Version 4 TCP IPv4 Properties 7 6 Section 7 Remote Interfaces 7 2 2 Windows Vista Settings 1 From the Windows Start menu right click on Network then select the Properties menu option The PC displays the Network and Sharing Center 2 Under Tasks select Manage Network Connections 3 Right click on the Local Area Connection icon and select the Properties option After you provide permission to continue the PC displays the Local Area Connection Properties window Figure 7 1 4 Select Internet Protocol Version 4 TCP IPv4 then click the Properties button The PC displays the Internet Protocol Version 4 TCP IPv4 Properties window Figure 7 2 If the Internet Protocol Version 4 TCP IPv4 option is not listed click the Install button to add it If assistance is needed press the F1 keyboard button to display Windows Vista Help The following steps
60. RADIENT OPTIMIZER 6 3 3 window When you accept the data the system will generate a gradient curve to separate the target compound from the nearest impurity When using optimized gradients PeakTrak may automatically insert an isocratic hold to further increase the resolution 6 4 6 Real time Gradient Editing The gradient shape can be changed during the run Any time after the fraction collector has positioned the drop former over the first tube click and drag the points on the gradient plot area of the Main window You can also add or remove points by clicking the Insert or Delete point buttons The Insert button allows you to add a single point when you click on the gradient profile Repeat this action to add more points The Delete button allows you to remove a single point when you click on it Note that only the portion of the gradient that has not yet occurred during the run can be modified 6 44 Section 6 User Help Reference 6 4 7 Bypassing the Solid Load Cartridge Rf 200 Rf 200i and Rf 4x Only Occasionally impurities precipitate in the solid load cartridge as compounds are purified This may cause high back pressures resulting in reduced flow rates and long run times Clicking this button changes the injection valve position which removes the solid load cartridge from the solvent path and relieves the back pressure CAUTION Bypassing the solid load cartridge too soon during the purification run may leave desired
61. RING THE WAVELENGTH RATIO 6 4 8 topic for more details on this feature Rf 75 and Rf 150 systems have a fixed UV detection wavelength of 254 nm The peak detection settings for these systems include slope based and peak width options These are described in A1 and A2 above Rf 75 and Rf 150 systems with the variable wavelength option include a control to type or set the detection wavelength 6 3 3 Gradient Optimizer Use this window to automatically generate a gradient curve from the results of two TLC plates Follow the on screen instructions then enter valid TLC data in the Gradient Optimizer window Click the OK button and the Method Editor window will display the new gradient curve 6 21 CombiFlash Rf User Manual When using the Gradient Optimizer the system may insert an isocratic hold into the generated gradient Furthermore the system may automatically extend the isocratic hold during the run to increase peak resolution To open the Gradient Optimizer window open the METHOD EDITOR 6 3 2 window then click the Gradient Optimizer button 6 3 4 Column Data Rf200 Rf200i and Rf4X systems The Column Data window reports information about the RediSep Rf column installed in the system Compatible RediSep Rf and RediSep Rf Gold columns have an RFID tag that identifies the column type The system reads this information to identify the size and media then automatically adjusts the default method to match the inserted c
62. Rf system It also allows you to configure the system for network connections for remote access and printing Open the Configuration window by selecting the TOOLS gt MANUAL CONTROL 6 2 3 2 menu command The configuration window displays three tabs INSTRUMENT CONFIGURATION 6 3 10 1 NETWORK CONFIGURATION 6 3 10 2 and USER PREFERENCES 6 3 10 3 6 3 10 1 Instrument Configuration Solvent Controls Solvent A1 through B2 From the list of solvents specify the solvent connected to the each port of the system Add Solvent button Click this button to enter a solvent name The system can store up to 16 solvent names For assistance in selecting appropriate solvents see the SOLVENT SELECTION oN LINE table Remove Solvent button Click this button to open a list of all entered solvent names Select the names you want to remove and then click the OK button Enable Solvent Level Sensing Rf 200 Rf 200i and Rf 4x systems If this option is selected the system will check the solvent levels before and during a run The system will alert you if a solvent level becomes too low Note that the solvent level monitoring option does not control the operation of Waste Solvent Level monitoring Waste level sensing is always enabled at the beginning of a run Minimum Solvent Level This value is density dependent thus a range is given To understand the range consider that dichloromethane is more dense than hexane If
63. System Specifications 1 Continued UV Detection Wavelength 200 to 360 nm optional 200 to 780 nm UV Vis Monitor Wavelength Optional 200 to 360 nm optional with Purity Ratio 200 to 780 nm UV Vis Wavelength Accuracy 5 nm Fraction Accuracy 2mL flow rate 60 Electrical Safety per EN 61010 1 Pollution Degree 2 Installation Category II Maximum Altitude 2000 meters Note 1 All specifications are subject to change Note 2 For Rf200 columns undetected by RFID the max pressure is limited to 50 psi 1 8 Section 1 Introduction Table 1 2 CombiFlash Rf 200i Dimensions 1 Specifications 61 cm 24 x36 cm 14 1 x43 cm 17 HxWxD Weight 30 75 kg 67 8 lbs Power Options Input voltage range from 100 to 240 VAC 50 60 Hz 2 0 Amps system maximum Line cord is the disconnect device Line Frequency 50 60 Hz Ambient Temperature Humidity when connected to power 20 to 40 C maximum temperature must be at least 15 C below the boiling point of the solvent 0 to 95 non condensing Flow Rate Range 5 to 200 mL min Flow Rate Accuracy 5 Pressure Limit 2 1379 kPa 200 psi Pressure Accuracy 5 of full scale Gradient Formation Peak Detection Modes Binary gradient from A1 A2 and B1 B2 solvent inlets Solvent selection can be automatically controlled by the method Slope or threshold
64. The CombiFlash Rf system is available in different configurations Figure 1 1 CombiFlash Rf 150 system 10 100 mL min 150 psi without injection valve CombiFlash Rf 150 For general purpose Flash chromatog raphy needs the CombiFlash Rf can be factory configured as a 10 100 mL min 150 psi system that pumps two sol vents to form a binary gradient This configuration has an easily accessible fluid path that supports liquid or solid sample introduction and a fixed wavelength UV detector 254 nm Optionally the system can be configured for full spectrum 200 360 nm UV detection with secondary monitoring manual solvent selection valves to form a binary gradient from two of four solvent inlets and a 4 port manual injection valve 1 2 Section 1 Introduction Figure 1 2 CombiFlash Rf 200 system 5 200 mL min 200 psi with automatic injection valve CombiFlash Rf 200 For more advanced needs the Combi Flash Rf can be factory configured as a 5 200 mL min delivery flow rate 200 psi system that forms a binary gra dient by automatically selecting two solvents from four inlets This configuration has an automatic self cleaning injection valve for sample introduction and full spectrum UV 200 360 nm or UV vis 200 780 nm detection with secondary wavelength monitoring An internal air pump is used for post run column air purging active solvent level sensing and waste full bottle detection 1 3 CombiFla
65. To configure the system open the Configuration window by selecting Tools Configuration from the PeakTrak menu The configuration window has three tabs Instrument Configuration Network Configuration and User Preferences Network Configuration settings are discussed later in Section 7 REMOTE INTERFACES Adjust the following sections 3 10 1 through 3 10 6 on the Instrument Configuration and User Preferences tabs for the desired operation and then click OK to save the settings Some configuration settings do not take effect until the system is restarted After completing sections 3 10 1 through 3 10 6 follow the instructions in section 3 10 7 3 12 Section 3 Preparation Rf 150 Systems 3 10 1 Solvents When you installed the system up to two solvents were connected to the back panel This section of the Instrument Configuration tab allows you to name these solvents To do so The system is shipped with common solvent names already loaded If your solvent names are listed in the solvent drop down list boxes on the main window skip these steps 1 Click the Add Solvent button 2 Enter the solvent name and click the OK button 3 Repeat steps 1 and 2 for both solvents 3 10 2 General Settings CombiFlash Rf Name optional Use this option to name your system The name will appear in operational displays and run summaries This feature is useful when your laboratory has more than one CombiFlash Rf system Time
66. VENT B ethyl acetate PRIME B PERCENTAGE SOLVENT B 50 E FLOW RATE ml min PRIME 5096 B STOP Ly PUMP INTO TUBE e VALVE POSITION Through column iil DE a Figure 8 6 Manual Control Settings Quick Cleaning 5 Select Through column for the Valve Position option 6 Select Next for the Pump into Tube option By pumping into a collection tube the diverter valve is also cleaned of any residue during this operation 1 Click the Prime B button to pump 100 Solvent B through the bypass tube and into the collection tubes M Note Pumping solvent B at a moderate flow rate 25 to 45 mL min over time will usually solubilize obstructions Generally the recommended solvent is the highest polarity solvent you have recently used solvent B 8 Monitor the Raw Lamp Energy gauge As the system pumps solvent the raw lamp energy should gradually improve Pump solvent for two to five minutes or until the indicator reaches the far right of the green range When the indicator is in the yellow the lamp energy and flow cell are ready for the next run However continuing the cleaning for greater raw lamp energy should reduce the frequency of flow cell cleaning messages Figure 8 4 If after five minutes the indicator is still in the red range repeat the cleaning steps using an alternative solvent Table 8 1 Or complete the steps in the Monthly Flow Cell Cleaning procedure section 8 5 3 8 8 Section
67. a connection to a printer on the network Network settings are discussed in Section 7 REMOTE INTERFACES Do not select this option for these initial installation steps Default Run Units Run units are displayed along the X axis of the chromatogram Select Time in minutes or Column Volumes A column volume unit is the time it takes to pump enough solvent to exchange the volume held by the column The duration in minutes will vary according to the column media and size and the flow rate Gradient Method Select the standard gradient type Figure 3 8 Linear The system gradually mixes solvents A and B from the last programmed B until it reaches the next programmed B This method connects the inflection points that define the gradient with a straight line Step The system holds each B value until it reaches the next programmed B Because the system holds the B values the changes at each new value are nearly instantaneous producing a stepped appearance on the plot area Isocratic The system holds the initial solvent mix for the entire run Linear Step Isocratic Figure 3 8 Gradient types 3 15 CombiFlash Rf User Manual Original method Extension 2 Final Extension Enable Run Length Extension When enabled this option automatically extends the run if a peak is eluting at the end of the maximum B gradient This ensures that a late eluting peak fully comes off the column and is collected
68. am mid run solvent B changes by clicking the Edit Gradient button to open the GRADIENT TABLE Edit Gradient Although the gradient may be edited directly within the plot area you can also edit the gradient in tabular form Click this button to open the 6 17 CombiFlash Rf User Manual GRADIENT TABLE On Rf 200 Rf 200i and Rf 4x systems the Gradient table also lets you program solvent changes in the gradient For complete instructions on defining gradient curves see DEFINING A GRADIENT 6 4 5 Gradient Table The gradient table depicts the points that define the gradient curve in a tabular format From this table you can change Solvent B Rf 200 Rf 200i and Rf 4x systems the duration and B concentration of any point on the curve To do so select the new value for a gradient point Changes that you make to the table will be reflected in the gradient plot area You can also change the number of points on the curve by inserting or deleting rows in the table To change the number of points first highlight a table cell or row Then click the appropriate Gradient Button for the action you desire Insert Point This command or button will insert a row below the selected point Delete Point This command will delete the selected row Note that you cannot delete the initial point The system automatically updates the Run Length setting or scales the other points when you change the number of points and thei
69. ample and observe the flow rate from the Status Bar in the upper right corner of the main window As long as the system is delivering the programmed flow rate there is no need to bypass the cartridge 4 If the flow rate is slower than that programmed note the slowest displayed flow rate Use the equation below to determine if the cartridge can be bypassed T Cartridge Volume Rf Flow Rate 5 12 Section 5 Operation Where T is the time in minutes to wait after the system reaches the B solvent used to run the TLC plate Rf is the retention factor measured in step 1 Cartridge Volume is the volume in mL from step 2 and Flow Rate is the current flow rate in mL min 5 Observe the current B concentration in the status bar Once the system reaches the B percentage used for the TLC wait at least the time established as T After this time has elapsed it is likely that the desired compound has come off the cartridge and you can safely click the Cartridge Bypass button For example a TLC plate run at 30 B revealed that the last desired compound had an Rf factor of 0 35 An empty 25 gram size solid load cartridge was filled with sample adsorbed onto 10 grams of silica and then placed on the system The programmed flow rate was 60 mL min During the run the flow rate dropped to 30 mL min The equation is then T 45 mL 0 35 30 mL min 4 3 minutes In this example the Cartridge Bypass feature was used 4 3 minutes aft
70. an find the MAC Address on the Help gt About PeakTrak window 5 Click the OK button to save the settings 6 Confirm that your network administrator has completed any necessary network changes to support the system 7 Locate the CAT5 connection cable P N 480 6545 01 in the accessory package Insert one end into the Ethernet port on the back panel Connect the other end of the cable to your network access port You should be able to connect to the CombiFlash Rf from a PC on the network Section 7 1 1 and configure other network features such as network printing Section 7 1 2 and network file saving Section 7 1 3 Networks with Wi Fi access points may also support remote control with mobile digital devices Section 7 1 4 7 1 1 Network PC Access A successfully networked Combiflash Rf system can be accessed by a PC that meets the recommendations listed in Table 7 1 Table 7 1 Personal Computer Recommendations Operating Systems Microsoft Windows 2000 XP Vista or Windows 7 Hardware The computer hardware must meet the minimum required specifications of the selected operating System Network Protocol TCP IP Network Connection IEEE 802 3 Ethernet Display 800 x 600 pixels 256 colors minimum Internet Browser Last two versions of Microsoft Internet Explorer a This table shows supported PC configurations Other configurations or AJAX compatible browsers may be possible but are unsupported
71. ary wavelength detection When enabled type or select the peak detection wavelength in nanometers then click the Details button to configure additional settings described below Slope Based Select this option to enable Slope based peak detection When enabled peaks will be recorded if the slope algorithm indicates a peak within the Peak Width setting Peak Width Select the average peak width setting Peak widths are measured at the baseline The slope detector will detect peak widths ranging from about 0 2 to 2 times the peak width setting For example if you entered a peak width of 1 minute the range would be 12 seconds to 2 minutes For best operation the peak width should be set to just over the average peak width being separated For instance if the average peak width is 45 seconds you should enter a peak width of 1 minute For most flash chromatography 1 minute is a good starting point for a peak width Threshold Detection Select this option to enable Threshold peak detection When enabled peaks will be recorded if the Absorbance Units AU value is exceeded Threshold Type or select the Absorbance Units value to be used for Threshold detection Note If both Slope and Threshold peak detection methods are checked the system considers a peak to be present when any one condition is met This logical OR operator means that the system will cut a peak when either the Slope condition is true or when t
72. ased and peak width options described in A1 and 42 above The system will use its internal algorithms to cut detect and cut peaks based on the analog input signal Refer to the External Detector instruction sheet for cable and 6 20 Section 6 User Help Reference plumbing requirements The external detector option is not available on Rf 150 and Rf 200i systems ELS Detector Enable this option to use the evaporative light scattering ELS detector on an Rf 200i system When enabled click the Details button to configure additional settings Signal Gain You may modify the gain to scale the detector signal Slope Based and Threshold Refer to these features described in A1 and 42 above Spray Chamber Temperature The default settings are 50 C normal phase default methods and 15 C reverse phase This setting may be adjusted from 10 to 60 C so that detection can be optimized for the solvent system in use However the lower limit is 5 C below the ambient temperature Drift Tube Temperature The default settings are 55 C normal phase default methods and 60 C reverse phase This setting may be adjusted from 30 to 90 C but is limited to a range of 5 C below and 60 C above the ambient temperature Show Ratio If using two of the above peak detection options check this option to display a ratio of the selected wavelengths The ratio trace is often a useful indicator of purity Refer to the MONITO
73. ate static electricity CombiFlash Rf User Manual To Solvent Supply r To Solvent Level Sensing Connection 60 0923 017 Ferrule 60 0923 015 Compression Fitting Nut 60 5234 130 Label 1 of 4 60 5235 050 38 mm Cap or 60 5235 051 45 mm Cap 60 5234 129 Inlet Filter Assembly Place weighted filter at bottom of container Figure 2 2 Recommended solvent connection NCAUTION To prevent damage or premature wear to the pump and internal valves clean solvent should be used The solvent should not contain any dissolved solids 2 4 Section 2 Preparation Rf 200 and Rf 200i Systems NCAUTION If solvent containers are reused particulate matter can accumulate in the container Containers should be thoroughly cleaned before each refill Failure to clean the containers may result in damage to the solvent selection valve and internal components The CombiFlash Rf 200 and Rf 200i systems have four solvent inlet ports on the back panel two A solvents and two B solvents The system requires at least one and one B solvent to form a binary gradient and assumes that the B solvent is stronger than A The second set of inputs allow you to form alternative binary gradients or to change the B solvent mid run The accessory package includes several pieces to complete the solvent inlet connections To connect the solvent lines 1 Solvent lines between the check valves and the solven
74. ation when the PeakTrak screen is displayed Section 4 Preparation Optional 4x Module 4 10 Prime the Solvent Lines The Rf 4x module was factory tested with 1 1 water isopropyl alcohol and subsequently purged with air If residual fluids would affect your chromatography prime all channels on the system Channel 1 should have been primed during the installation steps in Section 2 13 page 2 21 Channel 1 can be primed using the Auto Prime menu command Channels 2 through 4 must be primed using the Manual Control menu command To do so 1 Ensure that the solvent containers are filled 2 Insert a priming tube assembly from the accessory package between the upper and lower column mounts on channels 2 through 4 3 Select the Tools Manual Control menu command 4 Select the Column 2 the channel option 5 Click the Prime B button The system pumps 100 solvent B through channel 2 When priming the first time inspect the solvent and waste connections to the system If any leaks are observed click the Cancel button to stop the pump Correct the leak by tightening the fitting an additional 1 4 turn and then restart the prime 6 Allow the system to pump solvent B for at least 3 minutes Then click the Cancel button 1 Click the Prime A button The system pumps 100 solvent A through channel 2 8 Allow the system to pump solvent A for at least 3 minutes Then click the Cancel button M Note This B A order
75. b lpO Network File Save Configuration When configured this feature allows the system to access a network directory from which it can save run files as PDF text or run monitor To enable this feature select a file type to be saved and enter the remaining settings necessary for network access Your network administrator will be able to provide the required settings NOTE All server and domain names must be fully qualified That is entries must include the full name server name domain domain Use forward slashes not back slashes when entering the network share path 6 3 10 3 User Preferences Language Select the language for PeakTrak Automatically Print Report at End of Run If this option is enabled the system will print a completed run report using the NETWORK PRINTING feature Default Run Units Run units are displayed along the X axis of the chromatogram Select Time Minutes or COLUMN VOLUMES oN LINE CV run units to be displayed on program settings and chromatogram displays The duration in minutes will vary according to the column media and size and the flow rate Enable Run Length Extensions When enabled this option automatically extends the run if a peak is eluting at the end of the programmed run length This ensures that a late eluting peak is still collected 6 31 CombiFlash Rf User Manual Gradient Method Select the desired gradient method option If the Linear method is selected solven
76. basic A dish type detergent solution can wash through particulates not picked up by the acidic solution The emulsification effect of detergent is an effective tool as well to help wash through oily samples Follow the detergent with several minutes of clean distilled water a Recent synthesis solvents should also be considered CombiFlash Rf User Manual 8 5 3 Monthly Flow Cell Cleaning Perform this procedure as part of your scheduled preventive maintenance or when Quick Cleaning As Required section 8 5 2 does not improve the lamp energy 1 Remove the column and insert a bypass tube between the upper and lower column mounts bypass tube 209 0165 46 can be found in the accessory kit 60 5239 006 2 From the menu select Tools Manual Control This opens the Manual Control window 3 Set the Flow Rate to 40 mL min Figure 8 6 4 Select Through column for the Valve Position option 5 Select Next for the Pump into Tube option 6 Place the B1 Solvent inlet line into a reservoir of methanol acetone or a strong solvent that readily dissolves residual sample material 1 Click the Prime B button to pump 100 Solvent B through the bypass tube and into the collection tubes 8 After three minutes click the Stop button Allow the system to stand for at least six hours Overnight is recommended 9 Return the B1 solvent line to the original solvent container 10 Perform the Quick Cleaning as Require
77. change the local area connection settings If the PC must be restored to its original network configuration record the present Internet Protocol TCP IP settings now 5 Select the Use the following IP address option 6 Enter the IP address 192 168 1 200 7 Enter the Subnet Mask 255 255 255 0 8 Click the OK button to close the Internet Protocol Version 4 TCP IPv4 Properties window Click the OK button to close the Local Area Connection properties window 9 Shut down the PC and place the CombiFlash Rf system in Standby CombiFlash Rf User Manual 7 2 3 Windows XP Settings 1 From the Windows Start button open the Control Panel 2 Locate the Network Connection icon and open this control panel 3 Highlight the LAN or High speed Internet Connection icon Select the Change settings of this connection option or right click and select Properties Refer to Figure 7 3 4 Highlight the Internet Protocol TCP IP item in the list and click on the Properties button Refer to Figure 7 4 5 Select the Alternate Configuration tab Click the User Configured option See Figure 7 5 6 Enter the IP address 192 168 1 200 7 Enter the Subnet Mask 255 255 255 0 8 Click the OK button to close the Internet Protocol properties window Click the Close button to close the Local Area Connection properties window 9 Shut down the PC and place the CombiFlash Rf system
78. channels 2 through 4 for automatic injection valve cleaning The Rf 4x module can automatically detect full waste bottles and stop operation to prevent overfilling a waste container This safety feature requires five air line connections one Waste Level Sense From Rf inlet to receive air or external gas from the CombiFlash Rf system four Waste Level Sense outlets 1 2 3 and 4 M Note The following instructions assume that the waste from each channel will be isolated in separate containers 4 4 Section 4 Preparation Optional 4x Module WASTE LEVEL SENSE FROM RF WASTE LEVEL SENSE VALVE WASTE OUT COLUMN e LO O From DISCONNECT SUPPLY RE BEFORE SERVICING MAINS 2 10O A B 3 Figure 4 2 Rf 4x Module back panel connections IW Note If the waste is to be collected in a common container skip the Column Waste and Waste Level Sense connections steps 1 through 7 and complete step 8 to route the Valve Waste Out from the Rf 4x module to the common container The waste tubing assembly connected to the back panel of the CombiF ash Rf system will route all waste fluids to the common collection container and sense the level To complete the waste line connections 1 At the waste container for the CombiFlash Rf system Channel 1 remove the black tubing that delivers Diverter Valve Waste fluid and the natural tubing that delivers Waste Level Sense air The cap and black tubin
79. chemists Stop and then Fast Forward the run after the last compound elutes This action skips the post separation flush If any of the above conditions appear consider allowing some of the runs to continue through the 8 5 CombiFlash Rf User Manual flush or run a high percentage of B solvent through the system for a few minutes at the end of each day If the separation runs always continue through the flush and the conditions still occur frequently edit the default column methods to extend the flush duration NCAUTION When using methanol as solvent B with silica column media do not pump more than 5096 solvent B Higher percentages of methanol might break down the silica structure possibly causing obstructions in the flow path 8 5 2 Quick Cleaning when Recommended When the lamp energy is lower than normal the system will recommend flow cell cleaning Figure 8 4 before starting a separation run CONTINUE AND CANCEP RN COLLECT ALL ea Figure 8 4 Flow cell cleaning message When the system displays this message you can Cancel Run recommended Click the Cancel Run button so you can perform a quick cleaning described in the following steps Continue Collect All Click this button to ignore the message Because the peak detection operation might be impaired the system automatically collects all fluids to avoid diverting compounds of interest to waste Help Click this button to display the flo
80. compounds on the solid load cartridge After removing the cartridge from the fluid path these compounds cannot be purified If you need to use this feature perform the following to ensure that you do not leave desired compounds on the cartridge 1 Run a TLC plate prior to starting the run Measure the Rf retention factor of the slowest eluting compound of interest The Rf should be between 0 2 and 0 8 the same range used by the GRADIENT OPTIMIZER 6 3 3 Note the Percent B solvent used to run the TLC plate If a compound displays tailing peaks as streaks on TLC the Rf used corresponds to the bottom of the streak 2 Load the Sample in the Solid Load Cartridge and note the cartridge size Refer to list below to determine the cartridge volume If the cartridge is partially filled with silica use the cartridge volume as if the cartridge were completely filled with silica e 5 gram cartridge 10 mL volume 25 gram cartridge 45 mL volume 65 gram cartridge 110 mL volume 3 Run the sample and observe the flow rate from the Status Bar in the upper right corner of the main window As long as the system is delivering the programmed flow rate there is no need to bypass the cartridge 4 If the flow rate is slower than that programmed note the slowest displayed flow rate Use the equation below to determine if the cartridge can be bypassed T Cartridge Volume Rf Flow Rate 6 45 CombiFlash Rf User Manual Wh
81. conditions that will drive off the solvent without affecting the compounds of interest These compounds remain bound to the media Once dried pour the media sample mixture into an empty cartridge 5 1 CombiFlash Rf User Manual To prepare an empty solid sample cartridge 1 Ensure the empty cartridge has a bottom frit CAUTION Missing frits may cause equipment damage UV detection problems or increased maintenance Frits prevent solids from entering the fluid path 2 Prepare the media and sample a Dissolve your sample in a minimal amount of a suitable volatile solvent b Place the media into the solvent If using silica gel a particle size of 40 60 pm 240 400 mesh is recommended The amount of silica required is about four to five times the mass of your sample c Agitate the solvent for a moment to allow the sample to adsorb to the silica d Remove the solvent with a suitable method such as rotary evaporation Alternatively you can dry the prepared cartridge after step 5 by drawing the solvent out using a vacuum or using the Rf 200 or Rf 200i system s air purge 3 Load the media and sample mixture into the cartridge Tap the cartridge on the benchtop to settle the mixture 4 Place a frit on the top of the cartridge Force the frit down against the mixture using the plastic plunger 5 Wipe any residual powder inside the neck of the cartridge On Rf 200 Rf 200i and Rf 4x systems you can remo
82. d protect the next user from being sprayed by solvents left in the lines by pressing Fast Forward so the instrument purges itself and is safe for its next use Solid Sample Cartridge with air purge NOT enabled i e purge time set to zero in Method Editor If you suspend the run with the Stop button remove the column and install the Prime Tube in place of the column Manually purge the Sample Cartridge by pressing Tools gt Manual Control gt Air Purge Cartridge Repeat this procedure for Air Purge Column Liquid sample loaded onto column with air purge enabled If you suspend the run with the Stop button remove the column and install the Prime Tube in its place Use the Fast Forward button to ensure no sample is left in the flow cell and fraction collection lines possibly affecting subsequent separations 5 10 Section 5 Operation Liquid sample loaded onto column with air purge NOT enabled If you suspend the run with the Stop button remove the column and install the Prime Tube in its place Manually purge the liquid lines by pressing Tools gt Manual Control Air Purge Column Next Tube This button advances the fraction collector to the next tube position This allows you to conveniently collect eluate of interest in a new tube When you click the Next Tube button the system immediately marks the tube advance on the chromatogram However the system delays the movement of the drop former over the tube
83. d section 8 5 2 and monitor the Raw Lamp Energy If the lamp energy is in the green or yellow range return the system to operation If the lamp energy is red contact Teledyne Isco s Technical Service department for assis tance 8 6 Injection Valve Maintenance CombiFlash Rf200 Rf 200i and Rf 4x systems have an automatic injection valve Under normal operating conditions this self cleaning valve does not require any maintenance If maintenance is required refer to Instruction Sheet 60 5232 475 CombiFlash Rf Injection Valve Replacement 8 10 Section 8 Maintenance 8 7 Electrical Troubleshooting If your instrument stops working and the touch panel display is off check the line cord connection If the line cord is connected properly check the fuses in the power input module located on the system s rear panel These are time delay fuses T type and rated according to the mains input power voltage CombiF lash Rf 200 and Rf 150 systems 100 to 129 VAC mains input 2 0 A fuse one required e 215 to 264 VAC mains input 1 0 A fuse two required CombiF lash Rf 200i system 100 to 129 VAC mains input 3 0 A fuse one required 215 to 264 VAC mains input 3 0 A fuse two required CAUTION Always replace fuses with the same type and rating The CombiFlash Rf 4x module does not have a user accessible fuse CombiFlash Rf User Manual DECLARATION OF CONFORMITY ISM1 A Applicati
84. d an end time column volumes may also be used Note that all times are relative to start of the run just following the sample injection Then choose a peak collection mode If you select Peaks the module will collect fluid only when the time window is active and a peak is present If you select All the module will collect all fluid during the time window regardless of the peak state Define up to three time windows e Air Purge Rf 200 Rf 200i and Rf 4x systems Type or select the duration of a post run air purge in minutes An air purge clears the solvent from the column and the fluid path forcing any liquid out the waste port If you are disposing the column after each run a sufficient post run air purge ensures that it will be empty To disable the air purge enter a value of 0 zero A column can also be manually air purged See MANUAL CONTROL 6 3 9 6 3 2 3 Run Notes Use this text entry box to enter comments or notes for the run These comments will be saved with the run and will appear in TXT and PDF reports 6 16 Section 6 User Help Reference 6 3 2 4 Gradient Plot Area The gradient plot area depicts the current gradient profile that will be used for the run The X axis of the plot displays the run time in minutes or COLUMN VOLUMES oN LINE The X axis scale can be adjusted by typing or selecting the Run Length The left Y axis displays absorbance units and the right Y axis indicates the percentage of solvent
85. d from the column before the run Loading Type options for the Rf 150 system are Equilibration Select this option if you intend to equilibrate the column before introducing the liquid or solid sample No Equilibration Select this option to skip column equilibration 5 Review or update the Start Rack and Start Tube 6 Review the Minimum Run Requirements The system will report the estimated solvent volumes expected waste collection tube usage etc You can use this information to verify that there will be enough solvent to complete the run the waste volume does not exceed the collection container s capacity and whether or not more tubes will be required during the run The Rf 200 Rf 200i and Rf 4x systems estimate the solvent volumes by monitoring the solvent level in the container and the known usage rate The systems continue to refine this estimate during operation Solvent level sensing is not available on the Rf 150 system 5 7 CombiFlash Rf User Manual If the waste level sensing tube is inserted correctly into the waste container the Rf 200 Rf 200i and Rf 4x systems will automatically suspend operation before an overflow condition might exist To prevent the run from being suspended before completing the run ensure that the container will hold the expected waste volume Waste level sensing is not available on the Rf 150 system 7 Rf 200i only If the ELSD will be used for the run ensure tha
86. deficiencies before returning the instrument to operation CombiFlash Rf User Manual Figure 8 1 Collector tray drain hole Figure 8 2 Top shelf drain hole 8 4 Vacuum or pressurized air applied to the outlet end of the drain tube must exist at the collection tray drain hole Vacuum or pressurized air applied to the outlet end of the drain tube must exist at the top shelf drain hole 8 5 Flow Cell Cleaning 8 5 1 Post Separation Section 8 Maintenance As a preventive measure all default column methods finish the separation run with a high percentage of solvent B Figure 8 3 This brief time two to three column volumes of strong solvent flushes residual compounds from the column flow cell and internal tubing End of Separation Separation Z Gradient Time or Column Volumes 100 C C l 75 Qo i F g HF 2 C D i 50 se E 77 9 Ps lt 1 jmd 2 End of 25 w Run L 0L 0 E Figure 8 3 Default post separation column and flow cell flush Skipping the post separation flush may cause residual compounds to build up and crystallize which might result in cross contaminating later separation runs higher operating pressures reduced flow cell lamp energy a noisy baseline on the absorbance trace frequent messages recommending flow cell cleaning Figure 8 4 Typically
87. e Due to the high pressure that is possible during rapid equilibration this option may not be desirable for some column media or purification methods 3 10 7 Restart the system Some configuration settings do not take effect until the system is restarted After completing sections 3 10 1 through 3 10 6 restart the system 1 If the Configuration window is not closed already click the OK button to close it 2 Place the Power switch below the LCD display in the Off position and wait one minute or more N CAUTION Always wait at least one minute after placing the Power switch in the Off position Moving the switch to the Off position begins a power down sequence that takes up to one minute to complete If you move the switch to the ON position before the sequence is complete the system may not start up correctly A CAUTION Except during an emergency never remove AC power immediately after placing the Power switch in the Off position The power down sequence includes file maintenance that takes up to one minute to complete Interrupting the file maintenance may corrupt files causing abnormal operation or a complete system failure 3 After one minute or more place the Power switch in the On position 4 The system is ready for operation when the PeakTrak screen is displayed 3 17 CombiFlash Rf User Manual 3 11 Prime the Solvent Lines Before the first use the system should be primed Ensure that the solvent
88. e These corrupted files can cause abnormal operation or a complete system failure that requires service Unless power must be removed due to an emergency always wait at least one minute after placing the system in Standby before removing the AC mains power cord CombiFlash Rf User Manual 8 2 Preventive Maintenance The system requires preventive maintenance for safe and reliable operation Refer to the schedule below for the minimum periodic maintenance requirements As Needed Perform these tasks as conditions require Cleaning section 8 3 Quick flow cell cleaning when recommended by a system alert message section 8 5 2 Every Run Allow the separation run to finish with a high percentage of solvent B to flush residual compounds from the column internal tubing and flow cell Refer to section 8 5 1 Monthly Perform these tasks at least monthly more frequently if conditions warrant Tubing Inspection section 8 4 Collection rack and tray cleaning section 8 3 1 Monthly flow cell cleaning section 8 5 3 8 3 Cleaning To clean the exterior surfaces use a cleaning cloth dampened with a mixture of distilled water and a mild detergent Use isopropyl alcohol for tougher stains On printed areas such as labels avoid rubbing vigorously or using aggressive solvents like acetone Each will ruin the printed text NCAUTION Do not immerse the instrument in a water bath or subject it to a liq
89. e When you select this command the system opens the AUTO PRIME VALVE WASH 6 3 8 window 6 2 3 2 Manual Control Choose this command to open the MANUAL CONTROL 6 3 9 window From this window you can operate the system manually Manual control can assist with priming or purging the internal plumbing and aid system troubleshooting 6 2 3 3 Auto Zero Choose this command to zero the absorbance trace during a run 6 2 3 4 Edit Defaults Choose this command to open the METHOD EDITOR 6 3 2 window from which you can modify the default method files To prevent tampering with the default column programs access to this function can be password protected using the SET PASSWORD 6 2 3 6 menu command See Also EDITING A DEFAULT METHOD 6 4 4 6 2 3 5 Configuration Choose this command to open the CONFIGURATION 6 3 10 window From this window you specify the solvents used with the system set default volumes for collection tubes set the system date and time configure network settings efc 6 7 CombiFlash Rf User Manual 6 2 3 6 Set Password Choose this command to open the SET PASSWORD 6 3 11 window Passwords protect unauthorized modifications to Default methods User management system date and time system password Before any of the above can be accessed the user must enter a password Password protection for the default methods can be disabled so all users can modify default methods 6 2 3 7 User Manag
90. e following discussion If a pure compound is eluting the absorbance is linearly related to the concentration of the compound in the solvent If the compound absorbs differently at different wavelengths the absorbance at each wavelength may be different but still linearly related For example assume a compound eluting from the system has an absorbance equal to 2 times the concentration at 254 nm This same compound at 220 nm has an absorbance of 1 5 times the concentration The ratio of these signals will be 1 33 Since the relationship of absorbance to concentration is not variable the ratio remains steady while the concentration changes from the beginning to the end of the 6 46 Section 6 User Help Reference peak During the duration of the peak the ratio will be 1 33 and this constant value is displayed as a horizontal line Now assume a case where there is a second compound eluting only slightly shifted in time from the original compound It is possible that the detection absorbance trace alone would indicate a single valid chromatographic peak In reality itis a combination of two peaks By monitoring a second wavelength it may be possible to reveal the second compound Because of the slight shift in time and the different absorbance properties of the two compounds the changing ratio during the detected peak would reveal the impurity Therefore one can therefore assume that if the ratio is not constant for the entire dura
91. e optional valve kit the second set of inputs allows you to manually switch between alternative binary gradient solvent systems or to change the B solvent mid run 3 4 Section 3 Preparation Rf 150 Systems NCAUTION To prevent damage or premature wear to the pump and internal valves clean solvent should be used The solvent should not contain any dissolved solids An optional manual solvent selection valve kit may be added to provide inlets for two A solvents and two B solvents If using the optional valve kit connect the kit and solvent lines according to the instruction sheet Accessory package P N 60 5239 009 includes pieces to complete the solvent inlet connections 1 Locate a Solvent Inlet Filter Assembly P N 60 5234 193 in the accessory package Connect the tubing end to the A Solvent inlet Thread the nut into the Solvent A Inlet and finger tighten 2 Connect the second Solvent Inlet Filter assembly to the B Solvent inlet 3 Insert the filter end of each solvent inlet filter assembly into their respective solvent containers 4 The accessory package includes labels to easily identify the inlet lines Apply the A and B labels to the solvent lines above the solvent container openings NCAUTION If solvent containers are reused particulate matter can accumulate in the container Containers should be thoroughly cleaned before each refill Failure to clean the containers may result in damage to the solvent
92. e peak detection and collection settings for the run You can return to the rack and tube display by clicking the Display Rack button See Also PEAKTRAK MENU OPTIONS 6 2 e STARTING A RUN USING DEFAULT SETTINGS 6 4 1 REAL TIME GRADIENT EDITING 6 4 6 e METHOD EDITOR 6 3 2 for advanced functions 6 3 2 Method Editor The Method Editor window has several sections The BUTTON BAR 6 3 2 1 at the top of the window gives you quick access to file operations a GRADIENT OPTIMIZER 6 3 3 and COLUMN DATA 6 3 4 Rf200 Rf200i and Rf4X Below the button bar are the RUN SETTINGS 6 3 2 2 and the RUN NOTES 6 3 2 3 The lower part of the window contains the GRADIENT PLOT AREA 6 3 2 4 and settings for PEAK COLLECTION 6 3 2 5 and PEAK DETECTION 6 3 2 6 6 3 2 1 Button bar The Method Editor button bar contains the following New Rf200 Rf200i and Rf4X Click this button to open a new method file using the default method settings for the detected column media and size Open Click this button to open a method file stored on the system s internal hard drive Save Click this button to save any modifications to the current method file Note that if you attempt to save modifications to a default method file the FILES 6 3 6 window will open instead so you can rename the file This preserves the default method If you do wish to edit and save the system s default methods use the TOOLS gt EDIT DEFAULTS 6 2
93. e rapid equilibration select this option to equilibrate the column at a high flow rate M Note Due to the high pressure that is possible during rapid equilibration this option may not be desirable for some column media or purification methods 2 12 7 Restart the system Some configuration settings do not take effect until the system is restarted After completing sections 2 12 1 through 2 12 6 restart the system 1 If the Configuration window is not closed already click the OK button to close it 2 Place the Power switch below the LCD display in the Off position and wait one minute or more NCAUTION Always wait at least one minute after placing the Power switch in the Off position Moving the switch to the Off position begins a power down sequence that takes up to one minute to complete If you move the switch to the ON position before the sequence is complete the system may not start up correctly NCAUTION Except during an emergency never remove AC power immediately after placing the Power switch in the Off position The power down sequence includes file maintenance that takes up to one minute to complete Interrupting the file maintenance may corrupt files causing abnormal operation or a complete system failure 2 20 Section 2 Preparation Rf 200 and Rf 200i Systems 3 After one minute or more place the Power switch in the On position 4 The system is ready for operation when the PeakTrak screen is d
94. e system will clean the injection valve Clicking the Fast Forward button will cause the system to skip any remaining time in the current step and advance to the beginning of the next step Unlike the Rewind button the Fast Forward button will not modify the current method s run length setting 6 3 1 4 Peak Collection Buttons Three Peak Collection buttons are located at the bottom of the Main window All Click this button to collect all fluids in the fraction collection tubes Peaks Click this button to collect only eluted peaks in the fraction collection tubes None Click this button to divert all fluids to the waste port 6 3 1 5 Peak Collection Data Peak Collection data is displayed in the Main window after a run You may also open a RUN FILE 6 1 3 for viewing data from previous runs The peak collection data is displayed on the left side of the Main window Rack and tube information Collected peaks are color coded in the rack diagram so that you can easily locate the peaks of interest The tube colors correspond to the color bars under the peaks displayed on the chromatogram If during the run more than one set of 6 13 CombiFlash Rf User Manual tube racks was filled use the Next and Previous Rack buttons to view the additional racks The table below the rack diagram displays the peak data in tabular form Display Method Click the Display Method button to view a summary of th
95. e system to be configured for network operation and a connection to a printer on the network Network settings are discussed in Section 7 REMOTE INTERFACES Do not select this option for these initial installation steps Default Run Units Run units are displayed along the X axis of the chromatogram Select Time in minutes or Column Volumes A column volume unit is the time it takes to pump enough solvent to exchange the volume held by the column The duration in minutes will vary according to the column media and size and the flow rate Gradient Method Select the standard gradient type Figure 2 8 Linear The system gradually mixes solvents A and B from the last programmed B until it reaches the next programmed B This method connects the inflection points that define the gradient with a straight line Step The system holds each B value until it reaches the next programmed B Because the system holds the B values the changes at each new value are nearly instantaneous producing a stepped appearance on the plot area Section 2 Preparation Rf 200 and Rf 200i Systems Isocratic The system holds the initial solvent mix for the entire run Linear Step Isocratic Figure 2 8 Gradient types Enable Run Length Extension When enabled this option automatically extends the run if a peak is eluting at the end of the maximum B gradient This ensures that a late eluting peak fully comes off the column
96. e with the run Solid Select this option if you have already prepared the sample placed it into a solid sample load cartridge and loaded the cartridge on the system You must also specify the cartridge type and size The system will automatically proceed until the end of the programmed run length 6 34 Section 6 User Help Reference e Liquid Select this option if you have prepared a liquid sample and plan to manually inject it into the injection port after column equilibration None on column Select this option if you have preloaded the sample on the column The system will skip column equilibration so that the sample will not be flushed from the column before the run Loading types for the Rf 75 system are Equilibration Select this option if you intend to equilibrate the column before introducing the liquid or solid sample No Equilibration Select this option to skip column equilibration 5 Review or update the Start Rack and Start Tube 6 Review the Minimum Run Requirements The system will report the estimated solvent volumes expected waste collection tube usage etc You can use this information to verify that there will be enough solvent to complete the run the waste volume does not exceed the collection container s capacity and whether or not more tubes will be required during the run Note The Rf 200 Rf 200i and Rf 4x systems estimate the solvent volumes by monitoring the solvent l
97. ecomes more pronounced as the ambient temperature increases 2 5 Connect Waste Lines NWARNING Risk of fire or equipment damage Failure to connect Waste Porttubing may allow organic solvents to pool in unsafe areas possibly creating dangerous levels of flammable vapors ANWARNING Risk of fire ignited by electrostatic discharges Never substitute the black tubing on CombiFlash systems The black tubing P N 023 0503 06 is conductive This tubing is required to dissipate static electricity NCAUTION Elevated flammable vapor levels are possible Ensure that the waste container is adequately ventilated preferably by placing it in a fume hood The system has two waste outlets on the back panel Diverter Valve waste and Inject Valve waste You can route waste to a common collection container or to individual containers The Waste Level Sense uses air pressure to detect the liquid level in the common waste container If using individual containers use the waste level sense to detect the level of the Diverter Valve waste container Under normal operating conditions it will fill faster than the Inject Valve waste container Empty both containers at the same time to prevent overfilling the unmonitored container 2 6 Section 2 Preparation Rf 200 and Rf 200i Systems To Diverter Valve Waste Connection To Waste Level Sense Connection m To Inject Valve Waste Connection 60 0923 017 Ferrule 60 0923 015 Compression Fitt
98. ection port above the column When you click the OK button the system equilibrates the column and then waits When instructed perform a liquid injection as described in steps a through d above or attach the solid sample cartridge to the injection port and connect the solvent delivery tubing Click the OK button to continue e Ifyou selected No Equilibration ensure that the solvent delivery tubing is connected The system starts the run immediately and assumes that the sample has been injected on the column or the solid load cartridge is in place 5 4 During the Run You may allow system to proceed with the run while monitoring the progress on the PeakTrak main window If desired gradient parameters may be modified during the run See the Real time Gradient Editing on line help topic As the run progresses the absorbance trace is drawn on the Gradient Plot area The separation or purification continues until the end of the run defined by the run length setting During the run data is automatically saved every few seconds PeakTrak automatically saves all run history on the system s internal hard drive in its native RUN format You can manually save a Run file as Text or PDF while stopped or any later time as long as the RUN file has not been deleted Selected files can be manually saved to a connected Flash drive To automatically save a Text or PDF file after each run configure the Network File Save feature see Section 7 1 3
99. ement Choose this command to open the USER MANAGEMENT 6 3 12 window Use this window to add or remove users from the system This feature allows you to create a working folder on the system for each user Use the FILE gt SET DATA PATH 6 2 1 4 command to select your folder To prevent unauthorized changes to user management this function is password protected 6 2 3 8 Calibrate Touchscreen Choose this command from the touch screen to calibrate the interaction between touch screen display elements and stylus location The calibration routine places a series of targets on the touch screen display Touch the exact center of each target with a stylus The routine will move clockwise through 20 calibration targets after which the system will return to the normal display During calibration routine ensure that no part of your hand or anything other than the stylus touches the screen Note If you select this command from a remote connection the system will start the calibration routine Invoking this from a remote connection may be useful if the system is so mis calibrated that this menu command cannot be selected from the touch screen 6 8 Section 6 User Help Reference 6 2 4 Help PeakTrak s Help menu has the following options 6 2 4 1 Help Choose this command to view the Help documentation 6 2 4 2 Service Qualified service personnel use this command to access the system s service functions These service function
100. ems found it is likely that organic vapors are present in the ambi ent environment of your laboratory In this case increase the setting of the Vapor Limit Settings greater than 25 are appro priate for laboratory environments with a somewhat elevated background solvent vapor concentration representing a safe setting but one with a reduced margin for error 2 12 5 Set Default Tube Volumes Click this button to open a window from which you can set the default volume of the collection tubes You can increase or decrease these values to set the desired default volume collected in each container Just be sure you don t overflow the containers by exceeding their capacity The system will automatically advance to the next tube when this default volume is met in each tube Keep in mind that advanced method settings discussed in Section 5 and the on line help may override this default setting During a run 2 17 CombiFlash Rf User Manual other factors such as automatic tube advances for detected peaks will affect collected volume 2 12 6 User Preferences View the User Preferences tab to configure system operation for the current user New systems will only have one user account named common Language Select the user s desired language from the drop down list box Automatically Print Report at End of Run if this option is enabled the system will print a report at the end of each run This option requires th
101. er the solvent gradient reached 30 B The calculation is conservative but allows you to minimize the time needed to keep the solid load cartridge in the fluid path 5 13 CombiFlash Rf User Manual 5 4 3 About Solvent Level Detection When enabled in the TOOLS gt Configuration window CombiF lash Rf 200 Rf 200i and Rf 4x systems will estimate the solvent volume usage by measuring the solvent level in the container and the monitoring the usage rate The system continues to refine this estimate of available solvent during operation At the beginning of a run the system calculates the amount of solvent required to complete the run The system will alert you if it detects that there is not enough solvent at the start of the run Likewise the system will alert you during the run if the estimated usage rate or the measured level indicates that there is insufficient solvent Note that pauses and automatic run extensions during a run will increase the required solvent volume The options available on the alert message will vary depending on when the condition occurs before or during a run and which method measured level or estimated usage rate indicated the condition To clear the alert message the user must select one of the options below Continue Select this option after you have added more solvent to the container After selecting this option the system measures the solvent level again If the level is sufficient to
102. ere T is the time in minutes to wait after the system reaches the B solvent used to run the TLC plate Rf is the retention factor measured in step 1 Cartridge Volume is the volume in mL from step 2 and Flow Rate is the current flow rate in mL min 5 Observe the current B concentration in the status bar Once the system reaches the B percentage used for the TLC wait at least the time established as T After this time has elapsed it is likely that the desired compound has come off the cartridge and you can safely click the Cartridge Bypass button For example a TLC plate run at 30 B revealed that the last desired compound had an Rf factor of 0 35 An empty 25 gram size solid load cartridge was filled with sample adsorbed onto 10 grams of silica and then placed on the system The programmed flow rate was 60 mL min During the run the flow rate dropped to 30 mL min The equation is then T 2 45 mL 0 35 30 mL min 4 3 minutes In this example the Cartridge Bypass feature was used 4 3 minutes after the solvent gradient reached 30 B The calculation is conservative but allows you to minimize the time needed to keep the solid load cartridge in the fluid path 6 4 8 Monitoring the Wavelength Ratio When using two peak detection options such as A1 with 2 or All Wavelength Detection with External a ratio of the detection sources can also be displayed which at times can provide the best indication of compound purity Refer to th
103. ese features described in A1 and 42 above Spray Chamber Temperature The default settings are 50 C normal phase default methods and 15 C reverse phase This setting may be adjusted from 10 to 60 C so that detection can be optimized for the solvent system in use This setting is limited to a minimum of 5 C below the ambient temperature Drift Tube Temperature The default settings are 55 C normal phase default methods and 60 C reverse phase This setting may be adjusted from 30 to 90 C but is limited to a range of 5 C below and 60 C above the ambient temperature Show Ratio If using two of the above peak detection options check this option to display a ratio of the selected wavelengths The ratio trace is often a useful 6 41 CombiFlash Rf User Manual indicator of purity Refer to the MONITORING THE WAVELENGTH RATIO 6 4 8 topic for more details on this feature Rf 75 and Rf 150 systems have a fixed UV detection wavelength of 254 nm The peak detection settings for these systems include slope based and peak width options These are described in A1 and A2 above Rf 75 and Rf 150 systems with the variable wavelength option include a control to type or set the detection wavelength 6 4 2 3 Saving Changes to the Method File After you have edited the method file you can save the changes for future use Click the Save As button give the method file a descriptive name then click the Save butt
104. ethod File PeakTrak controls the separation runs performed by the system through a Method file The system uses the method file to direct the system operation after you click the Play button PeakTrak categorizes the method settings as Basic or Advanced Basic settings are the frequently used controls accessed through the MAIN WINDOW 6 3 1 Use the METHOD EDITOR 6 3 2 window to access the Advanced settings Method files are stored by the system and can be opened for review reuse or modification To open a method file use the FILE gt OPEN 6 2 1 2 menu command Method files can be transferred to the system from an external storage device using the FILE gt IMPORT METHOD 6 2 1 9 menu command Method files use an mtd filename extension 6 1 2 1 Default Methods With the exception of the Rf 150 when PeakTrak is started and every time you insert a RediSep Rf column the system loads a default method A default method contains Teledyne 6 2 Section 6 User Help Reference Isco s recommended basic and advanced settings for the use of RediSep Rf columns on the system There is a default method for each size and media type of RediSep Rf column See REDISEP RF COLUMN SELECTION GUIDE ON LINE Default methods provide a starting point for your separation or purification From these initial settings you can perform a purification run or you can modify the settings for your next run Subsequent runs will use the settings in the act
105. evel in the container and the known usage rate The system continues to refine this estimate during operation See also ABOUT SOLVENT LEVEL DETECTION oN LINE Note If the waste level sensing tube is inserted correctly into the waste container the system automatically will suspend operation before an overflow condition might exist To prevent the run from being suspended before completing the run ensure that the container will hold the expected waste volume Waste level sensing is not available on the Rf 75 system Rf 200i only If the ELSD will be used for the run ensure that the ELSD option is selected Otherwise you may disable the ELSD The Rf 200i system will automatically check this option if the previous run used the ELSD 6 35 CombiFlash Rf User Manual 8 Click OK and the run begins The system responds according to the sample Loading Type you selected in step 4 Rf 200 Rf 200i and Rf 4x systems If you selected Solid Pause the system equilibrates the column with the starting B and then waits while you prepare the solid sample load cartridge Place the cartridge on the system and click OK to continue If you selected Solid the system equilibrates the column and immediately starts the separation If you selected None on column the system skips the equilibration and immediately starts the separation If you selected Liquid the system equilibrates the column and then waits When instructed perform the f
106. f 200 only Vacuum or pressurized air applied to the outlet end of the drain tube must exist at the top shelf drain hole Figure 2 5 Top Shelf Drain 3 Route the end of both drain tubes to a suitable waste fluid collection container Section 2 Preparation Rf 200 and Rf 200i Systems It may be necessary to extend the drain tube If so splice the tubing with user supplied tubing The user supplied tubing should have an inside diameter no smaller than the existing drain tubing and must be compatible with the solvents used by the system Route this extension tubing to the waste collection vessel CombiFlash Rf 200i only The CombiFlash Rf 200i system has additional drains that must be connected 4 Fill a syringe with 1 mL of liquid such as water or isopropyl alcohol and inject it into the P Trap port 5 Locate the l4 inch I D silicone tubing part number 029 1312 02 and a nylon tubing clamp part number 149 2002 08 from the accessory kit Connect one end of the tubing to the P Trap port and secure it with the clamp 6 Route the end of the P Trap drain tube to a suitable waste fluid collection container M Note If accessible the outlet end of the P Trap drain tube may be used to refill the P Trap 7 Connect user supplied tubing 1 2 inch I D to the exhaust port and route the exhaust away from the system This will prevent unnecessary solvent vapor alarms M Note The exhaust gas is dry and does not require a
107. format 6 1 PeakTrak Overview PeakTrak is the control software for Teledyne Isco s CombiFlash Rf system This overview provides a description of The PEAKTRAK WINDOW ELEMENTS 6 1 1 The METHOD FILE 6 1 2 The RUN FILE 6 1 3 HOW TO GET PEAKTRAK HELP 6 1 4 6 1 1 PeakTrak Window Elements The MAIN WINDOW 6 3 1 displays the current METHOD FILE 6 1 2 From this window you can access many of PeakTrak s features view the system status and view or edit the method file settings The elements of the main window are the Menu The topmost item in the window is the PEAKTRAK MENU OPTIONS 6 2 from which you can access all of PeakTrak s features 6 1 CombiFlash Rf User Manual System Status PeakTrak displays the system status to the right of the current method Status messages may include e The system mode current position in a run expressed in time or column volumes current B peak status flow rate current time and date Main region This area is where you can view or modify method file settings Frequently used method settings are displayed on the MAIN WINDOW 6 3 1 Advanced method settings are displayed on the METHOD EDITOR 6 3 2 window Many of the PEAKTRAK WINDOWS 6 3 contain these main elements or a subset of them Note that the available commands and options will change according to the active window the state of the system and the current file 6 1 2 M
108. g from the Inject Valve Waste should remain attached to the waste bottle 2 At the CombiFlash Rf system back panel disconnect the black tubing from the Diverter Valve Waste port and the natural tubing from the Waste Level Sense port 4 5 CombiFlash Rf User Manual The Inject Valve Waste tubing must remain connected between the Rf system back panel and its waste container Refer to Figures 4 3 and 4 4 for steps 3 and 4 3 Locate the 20 inch 50 cm long Column Waste Assembly P N 60 5234 192 in accessory package 60 5239 008 Connect this tubing between the Combiflash Rf system and the Rf 4x module a Connect the cut end of the black tubing to the CombiFlash Rf system Diverter Valve Waste port Use a black nut and ferrule from the accessory package Finger tighten the nut to seat the ferrule in the bulkhead fitting oj Figure 4 3 Preparing a bulkhead fitting connection Ferrule Nut Tubing b Connect the other end of the black tubing to the Rf 4x port labeled From Rf in the block labeled Column Waste Finger tighten the threaded fitting 4 Locate the 20 inch 50 cm length of natural tubing from the accessory package Using nuts 60 0923 015 and ferrules 60 0923 017 connect this tubing between the CombiFlash Rf system s Level Sense port and the Rf 4x module s Waste Level Sense From Rf port Level Sense Waste Solvent air Figure 4 4 CombiFlash Rf system to Rf 4x waste tubing connec
109. gned hereby declare that the design of the equipment specified above conforms to the above Directive s and Standards as of April 12 2007 William Foster USA Representative Teledyne Isco Inc William Foster Director of Engineering Teledyne Isco Inc 4700 Superior Street Lincoln Nebraska 68504 Phone 402 464 0231 Fax 402 464 4543 60 5232 141 DECLARATION OF CONFORMITY I8M1 A Application of Council Directive Manufacturer s Name Manufacturer s Address Telephone Facsimile Equipment Type Environment Trade Name Model No Year of Issue Standards to which Conformity is 2004 108 EC The EMC Directive 2002 96 EC The WEEE Directive 2006 95 EC The Low Voltage Directive Teledyne Isco Inc 4700 Superior Lincoln Nebraska 68504 1398 USA P O Box 8253 Lincoln NE 68501 2531 1 402 464 0231 1 402 465 3799 Laboratory Equipment for Light Industrial Commercial Environments Rf200i 2012 EN 61010 1 2010 Safety Requirements for Electrical Equipment for Declared Measurement Control and Laboratory Use EN 61326 1 2006 EMC Requirements for Electrical Equipment for Measurement Control and Laboratory Use EMC Standard Description Severity Applied Performance Criteria EN 61000 4 2 Electrostatic Level 2 4kV contact discharge A Discharge Level 3 8kV air discharge EN 61000 4 3 Radiated RF 80 MHz to 2 7 GHz 80 AM a
110. he Threshold condition is true Monitor Enable this option to only use the detection source as a monitor When enabled the detection source will be displayed as a trace on the gradient plot area but will not be used to cut peaks 6 40 Section 6 User Help Reference e All Wavelength Detection Enable this option to detect peaks within a user selected range of wavelengths When enabled click the Details button to configure additional settings These settings include the slope based and peak width options described in 41 and 22 above Additionally you can type or select the minimum and maximum wavelengths limits in nanometers e External Detector Rf200 Rf200i and Rf4X systems only Enable this option to use a 0 to 1 volt analog input signal from an external detector When enabled click the Details button to configure additional settings These settings include the slope based and peak width options described in A1 and 42 above The system will use its internal algorithms to cut detect and cut peaks based on the analog input signal Refer to the External Detector instruction sheet for cable and plumbing requirements ELS Detector Enable this option to use the evaporative light scattering ELS detector on an Rf 200i system When enabled click the Details button to configure additional settings Signal Gain You may modify the gain to scale the detector signal Slope Based and Threshold Refer to th
111. he ambi ent environment of your laboratory In this case increase the setting of the Vapor Limit Settings greater than 25 are appro priate for laboratory environments with a somewhat elevated background solvent vapor concentration representing a safe setting but one with a reduced margin for error 3 10 5 Set Default Tube Volumes Click this button to open a window from which you can set the default volume of the collection tubes You can increase or decrease these values to set the desired default volume collected in each container Just be sure you don t overflow the containers by exceeding their capacity The system will automatically advance to the next tube when this default volume is met in each tube Keep in mind that advanced method settings discussed in Section 5 and the on line help may override this default setting During a run other factors such as automatic tube advances for detected peaks will affect collected volume 3 10 6 User Preferences View the User Preferences tab to configure system operation for the current user New systems will only have one user account named common Language Select the user s desired language from the drop down list box 3 14 Section 3 Preparation Rf 150 Systems Automatically Print Report at End of Run if this option is enabled the system will print a report at the end of each run This option requires the system to be configured for network operation and
112. hich you can set the default volume of the collection tubes You can increase or decrease these values to set the desired default volume collected in each container Be sure you do not overfill the containers by exceeding their capacity The system will automatically advance to the next tube when this default volume is met in each tube Keep in mind that the PEAK COLLECTION 6 3 2 5 settings on the METHOD EDITOR 6 3 2 window may override this default volume During a run other factors such as automatic tube advances for detected peaks will affect collected volume See also LOADING A COLLECTION RACK oN LINE 6 29 CombiFlash Rf User Manual Enable Automatic Column Purchase This feature requires a current subscription in a Teledyne Isco Column Plan Contact your sales representative for more information on this feature The certificate issued with the column plan includes instructions for activating this feature Multiple Wavelength Detection Rf 75 and Rf 150 Only This feature requires a valid activation code for the variable wavelength option Contact your sales representative for more information on this feature 6 3 10 2 Network Configuration Network configurations settings require assistance from your network administrator Contact your Information Technology department before changing the settings Instrument IP Address Configuration Use these settings to specify the instrument s network address and its parameters
113. hod Files 6 4 4 Editing a Default Method 6 4 5 Defining a Gradient 05 6 4 6 Real time Gradient Editing 6 4 7 Bypassing the Solid Load Cartridge 6 4 8 Monitoring the Wavelength Ratio 6 4 9 Viewing runs llle 6 4 10 Manual Control of the CombiFlash Rf 6 4 11 Setting the Vapor Limit 7 Remote Interfaces 7 1 Network Configuration 005 7 1 1 Network PC Access 0000005 CombiFlash Rf User Manual 7 1 2 Network Printing 04 7 3 7 1 3 Network File Save Configuration 7 3 7 1 4 Apple Mobile Digital Devices 7 4 7 2 Direct Connection 0 00 RR RR KK KK 7 4 7 2 1 Windows 7 Settings KK KK 7 5 7 2 2 Windows Vista Settings 7 7 7 2 3 Windows XP Settings 7 8 7 2 4 Windows 2000 Settings 7 10 7 2 5 Completing the Direct Connection 7 12 7 3 Direct Printing 0 0 0 eee 7 12 8 Maintenance 8 1 System Standby and Shut Down 8 1 8 2 Preventive Maintenance 0085 8 2 8 3 Cleanibg 2 3 02 dae DA oH Sen eae s 8 2 8 3 1 Collection Rack and Tray Cleaning 8 3 8 4 Tubing Inspection KK RR RR RR KK 8 3 8 5 Flow Cell Cleaning KK RR KK KK KK 8 5 8 5 1 Post Separation 00000 ee 8 5 8 5 2 Quick Cleaning when Recommended 8 6
114. ica gel columns e Part number 60 5237 051 Test kit containing four of the 25 gram size solid load cartridges pre filled with 0 5 grams of test sample NPHE and four 4 gram RediSep Rf silica gel columns The test kits include instructions 3 19 CombiFlash Rf User Manual 3 13 Installation Qualification Checklist Table 3 1 Installation Qualification Checklist Installer Operator Description Initials Initials 3 1 UNPACKING THE UNIT 3 2 INSTRUMENT LOCATION 3 3 CONNECT POWER 3 4 CONNECT SOLVENT LINES 3 5 CONNECT WASTE LINES 3 6 CONNECT AND ROUTE DRAIN LINES 3 7 POSITION THE SYSTEM 3 8 INSTALL COLLECTION TUBE RACKS 3 9 TURN ON POWER 3 10 CONFIGURE THE SYSTEM 3 11 RESTART THE SYSTEM 3 12 SYSTEM VERIFICATION Certification of Section 3 Completion Installer Name print Installer Signature Date Operator Name print Operator Signature Date Comments 3 20 CombiFlash Rf User Manual Section 4 Preparation Optional 4x Module This section provides instructions for unpacking and installing the Combiflash Rf 4x module after completing the CombiFlash Rf 200 or Rf 200i system installation Section 2 To prepare the system for operation sequentially follow all instructions in sections 4 1 through 4 10 Section 4 12 contains an Installation Qualification checklist If required sign off
115. ide panels If there is any damage to the shipping carton or any components contact the shipping agent and Teledyne Isco or its authorized representative immediately CombiFlash Rf User Manual NWARNING If there is any evidence that the system has been damaged in shipping do not plug it into AC power Contact Teledyne Isco or its authorized representative for advice Compare the contents of the boxes with the enclosed packing slips If there are any shortages contact Teledyne Isco immediately The fraction collector arm was stowed to prevent damage during shipping The arm stowing bracket should be removed at this time 1 Loosen the bracket s two thumbscrews along the sides of the arm Figure 2 1 Thumbscrew Thumbscrew Figure 2 1 Remove arm stowing bracket and spacer 2 Push the arm to the left or right remove the spacer 3 Store the bracket and spacer These parts should be reinstalled if the system must be shipped again 2 2 Instrument Location The CombiFlash Rf 200 and Rf 200i systems have a relatively small footprint requiring about 1550 square centimeters 240 in of level bench space Ensure that the system has at least 3 cm 1 25 of air space behind it for ventilation Additional space may be required for solvent and waste containers The optional 4x expansion module is installed to the left of the system This module requires an additional 933 cm 145 in 2 2
116. in Standby lni xl ge Figure 7 8 LAN or High speed Internet Connection 7 8 Section 7 Remote Interfaces 1 Local Area Connection Properties Internet Protocol TCP IP Properties Figure 7 5 Alternate Configuration Settings CombiFlash Rf User Manual 7 2 4 Windows 2000 Settings 1 From the Windows Start button select Settings gt Control Panel to view the control panels 2 Locate the Network and Dial up Connections icon and open this control panel 3 Highlight the Local Area Connection icon View the properties by right clicking and selecting Properties The Local Area Connection Properties Window will open Figure 7 6 4 Highlight the Internet Protocol TCP IP item in the list and click on the Properties button The Internet Protocol Properties Window will open Figure 7 7 The following steps change the local area connection settings If the PC must be restored to its original network configuration record the present Internet Protocol TCP IP settings now 5 Click the Use the following IP address option 6 Enter the IP address 192 168 1 200 7 Enter the default Subnet Mask 255 255 255 0 8 Click the OK button to close the Internet Protocol properties window Then click the OK button to close the Local Area Connection properties window 9 Shut down the PC and place the CombiFlash Rf system in Standby
117. ing Nut 60 5234 143 Waste Tubing Assembly Ventilation Hole Luer fitting If necessary connect user supplied tubing and See Top View route to fume hood Tubing Stop ferrule 60 5235 053 38 mm Waste Cap or 60 5235 054 45 mm Waste Cap about 2 inches 5 cm Top View Diverter Valve Waste Waste Level Sense O e Ventilation Inject Valve Waste Figure 2 8 Recommended waste connections with plastic plate 2 7 CombiFlash Rf User Manual The accessory package includes parts to complete the waste connections Figure 2 3 shows the recommended connections for user supplied waste containers with either a 38 or 45 mm opening GL 38 or GL 45 Refer to the steps in section 2 4 to connect the tubing to the system using the compression fittings Then complete the connections refer to Figure 2 3 1 Push the three tubes through the cap The tubing will fit snugly in the cap and may require some force 2 Push the Waste Level Sense tubing until the Tubing Stop is against the cap This ensures that the outlet is about two inches 5 cm below the container s maximum level Should the container not have a GL 38 or GL 45 opening omit the use of the waste cap Ensure the solvent tubes are secured so they drain into the waste container Secure the Waste Level Sense air tubing so its outlet is at least two inches 5 cm below the container s maximum level Seal the container opening to avoid solvent vapors
118. ired instrument to the customer Teledyne Isco will not pay air freight or customer s packing and crating charges This warranty does not cover loss damage or defects resulting from transportation between the customer s facility and the repair facility The warranty for any instrument is the one in effect on date of shipment The warranty period begins on the shipping date unless Teledy ne Isco agrees in writing to a different date Excluded from this warranty are normal wear expendable items such as charts ribbon lamps tubing and glassware fittings and wetted parts of valves and damage due to corrosion misuse accident or lack of proper maintenance This warranty does not cover products not sold under the Teledyne Isco trademark or for which any other warranty is specifically stated This warranty is expressly in lieu of all other warranties and obligations and Teledyne Isco specifically disclaims any warranty of merchantability or fitness for a particular purpose The warrantor is Teledyne Isco 4700 Superior Lincoln NE 68504 U S A This warranty applies to the USA and countries where Teledyne Isco does not have an authorized dealer Customers in countries outside the USA where Teledyne Isco has an authorized dealer should contact their Teledyne Isco dealer for warranty service Before returning any instrument for repair please call fax or e mail the Teledyne Isco Service Department for instructions Many problem
119. isplayed 2 13 Prime the Solvent Lines Before the first use the system should be primed Ensure that the solvent containers are filled then 1 Select the Tools gt Auto prime menu command 2 Select the desired A and B solvents from the drop down list boxes 3 Click the Play button to start priming the system The system pumps 100 Solvent B through the system and out the Waste port and then repeats with 100 Solvent A This B A order leaves the internal lines filled with solvent more appropriate for the beginning of a purification run When priming the first time inspect the solvent and waste connections to the system If any leaks are observed click the Cancel button to stop the Auto Prime Correct the leak by tightening the fitting an additional 1 4 turn and then restart the Auto Prime from step 1 To advance to the next Auto Prime step you can click the Fast Forward button The Auto Prime window closes when finished After Auto Priming the system is ready for operation Section 5 Use Auto Prime for quick solvent changes by pumping a fixed amount of solvent through the waste fluid path at 100 mL min For more advanced priming functions such as varying the flow rates fluid paths etc or to purge the system with air before changing between normal and reversed phase solvent systems use the Tools gt Manual Control option CombiFlash Rf User Manual 2 14 System Verification It is recommended that
120. ive window If you have modified the settings you can save the method file for future use If you find that the default methods are not a practical starting point for your applications the default methods can be changed to meet your specific requirements See EDITING A DEFAULT METHOD 6 4 4 6 1 3 Run File When the system has finished a run it saves the run history in a Run file The history includes the METHOD FILE 6 1 2 settings rack and tube information and a chromatogram You can open and review the Run files stored by the system To open a Run file use the FILE gt OPEN 6 2 1 2 menu command Run files use an run filename extension 6 1 4 How to get PeakTrak help PeakTrak help can be displayed by selecting Help Help from PeakTrak s menu If you need additional help refer to Teledyne Isco s CONTACT INFORMATION ON LINE 6 2 PeakTrak Menu Options PeakTrak menus include FILE 6 2 1 METHOD EDITOR 6 2 2 TOOLS 6 2 3 HELP 6 2 4 6 2 1 File The file menu lists the following commands 6 3 CombiFlash Rf User Manual 6 2 1 1 New Choose the New command to open a new method file Rf150 The main window opens using the currently selected method Rf200 and Rf200i PeakTrak will open the main window using the default program settings for the detected column size 6 2 1 2 Open Choose this command to open a METHOD FILE 6 1 2 mtd or a RUN FILE 6 1 3 run stored on the system
121. k drive 1 6 Section 1 Introduction 1 4 Specifications Table 1 1 CombiFlash Rf 150 and Rf 200 System 1 Specifications Rf 150 Systems Rf 200 Systems Without Inject Valve With Inject Valve Overall Dimensions 61 x 36 x 43 cm 24 x 14 1 x 17 HxWxD Weight 22 3 kg 49 2 Ibs 24 5 kg 54 Ibs Power Options Input voltage range from 100 to 240 VAC 50 60 Hz 2 0 Amps maximum Line cord is the disconnect device Line Frequency 50 60 Hz Ambient Temperature 20 to 40 C maximum temperature must be at least 15 C below the boiling point of the solvent Humidity when 95 relative humidity maximum at 20 to 40 C connected to power Flow Rate Range 10 to 100 mL min 5 to 200 mL min Flow Rate Accuracy 10 5 tested with water at 138 kPa or 20 psi Pressure Limit 2 Up to 1379 kPa 200 psi 1 034 kPa 150 psi column dependent Pressure Accuracy 10 of full scale 5 of full scale Gradient Formation Binary gradient from A1 Binary gradient from A1 and B1 solvent inlets A2 and B1 B2 solvent Optional manual valves inlets Solvent selection to select from A1 A2 can be automatically and B1 B2 inlets controlled by the method Gradient Accuracy 2 of full scale typical 1 of full scale 5 maximum Peak Detection Modes Slope or threshold Flow Cell Pathlength 0 1 mm 25 1 7 CombiFlash Rf User Manual Table 1 1 CombiFlash Rf 150 and Rf 200
122. k nut and ferrule in the accessory package Slide the nut and ferrule onto the tubing as shown in Figure 3 3 Ferrule Tubing Nut 3 6 Section 3 Preparation Rf 150 Systems Figure 3 3 Preparing a bulkhead fitting connection b Fully insert the tubing into the threaded bulkhead fitting on the back panel of the system c Finger tighten the nut onto the threaded bulkhead fitting This will seat the ferrule in the fitting 2 Route the tubing and insert the weighted end into the user supplied waste container 3 6 Connect and Route Drain Lines Z WARNING Risk of fire or equipment damage Failure to connect drain lines may allow organic solvents to pool in unsafe areas creating a potential for dangerous levels of flammable vapors Improper draining may damage the instrument s internal components 3 7 CombiFlash Rf User Manual The CombiFlash Rf 150 system has drain tubes extending from its top shelf and back panel The tubes drain away any liquid spilled on the top shelf and the tray beneath the fraction collection racks 1 Test the fraction collector drain by connecting a vacuum or air supply source to the outlet end of the drain tube Then verify the presence of such vacuum or air supply source on the drain Figure 3 4 Vacuum or pressurized air applied to the outlet end of the drain tube must exist at the collection tray drain hole Figure 3 4 Fraction Collect
123. l any run and view it from the system as long as the RUN file has not been deleted To save the post run report for viewing elsewhere you can save the run history as text or PDF files as needed This Disabled option is useful when there is no need or only an occasional need to save a run history file externally To save all run histories automatically as a text or PDF file configure the Network File Save option This allows the CombiFlash Rf system to access the corporate network and save the file in a selected Network Share folder Consult with your network administrator to configure these fields 7 3 CombiFlash Rf User Manual 7 1 4 Apple Mobile Digital Devices You can wirelessly monitor and control a CombiFlash Rf system using the PeakTrak app The app is designed for a mobile digital device iPod Touch iPhone and iPads running iOS 4 2 or later The PeakTrak app is available for download from the App Store Each CombiFlash Rf system requires an active license before using the PeakTrak app A license may be ordered from Teledyne Isco using part number 60 5237 073 which contains the necessary software on a USB flash drive Refer to the instructions supplied with 60 5237 073 to install the license key and configure the system and mobile digital devices to support remote control M Note Remote communication requires the CombiF ash Rf system and mobile digital device to share a common network with Wi Fi access in the remo
124. lash Rf User Manual the system is ready to perform a run using the default settings These default settings are preset to the most common configurations used by chemists and have been optimized for the use of RediSep Rf columns The default settings will collect all fluid in the fraction collector rack e differentiate between UV 254 nm vs non UV absorbing fluid and place the fluids into separate tubes cut UV absorbing peaks based on slope detection or level threshold To start the run from the PeakTrak Main window 1 Enter a sample name If you choose not to PeakTrak will enter the date and time as the sample name when you start the run 2 Review the PeakTrak Main window settings If you want to change any settings refer to the Editing a Method on line help topic 3 Click the Play button 4 Select the sample Loading Type from the list The Loading Type should support the sample injection technique you intend to use see SAMPLE INJECTION METHODS oN LINEI Possible sample loading types for the Rf 200 Rf 200i and Rf 4x systems are Solid Pause Select this option if you intend place the sample into a solid sample load cartridge but have not yet prepared the sample You must also specify the cartridge type and size When you click the run button the system will perform a column equilibration and then wait while you prepare the cartridge After you have placed the cartridge on the system click OK to continu
125. leaves the internal lines filled with solvent appropriate for the beginning of a purification run 9 Repeat steps 4 through 8 for channels 3 and 4 Be sure to change the channel option before starting the pumps CombiFlash Rf User Manual 4 11 System Verification It is recommended that the system operation be verified for each channel To do so use a pre packaged CombiFlash Rf test kit e Part number 60 5237 050 Test kit containing five of the 5 gram size solid load cartridges pre filled with 0 5 grams of test sample NPHE and five 4 gram RediSep Rf silica gel columns e Part number 60 5237 051 Test kit containing four of the 25 gram size solid load cartridges pre filled with 0 5 grams of test sample NPHE and four 4 gram RediSep Rf silica gel columns The test kits include instructions 4 14 Section 4 Preparation Optional 4x Module 4 12 Installation Qualification Checklist Table 4 1 Installation Qualification Checklist Installer Operator Description Initials Initials UNPACKING THE UNIT INSTRUMENT LOCATION CONNECT POWER CONNECT SOLVENT LINES CONNECT WASTE LINES CONTROL CONNECTION POSITION THE SYSTEM REMOVE SHIPPING HARDWARE TURN ON POWER PRIME THE SOLVENT LINES SYSTEM VERIFICATION Certification of Section 4 Completion Installer Name print Installer Signature Date Operator Name print Operator Signature Date
126. ludes self diagnostics 2 Monitor the movement of the fraction collector arm The arm should move to the left rear corner of the fraction collector area You should not hear any grinding noises during the homing sequence If grinding is heard there is a problem with the homing of the fraction collector arm 3 The system is ready for operation when the PeakTrak screen is displayed 2 12 Configure the System Before operating the system it should be configured for the desired operation To configure the system open the Configuration window by selecting Tools Configuration from the PeakTrak menu The configuration window has three tabs Instrument Configuration Network Configuration and User Preferences Network Configuration settings are discussed later in Section 7 REMOTE INTERFACES Adjust the following sections 2 12 1 through 2 12 6 on the Instrument Configuration and User Preferences tabs for the desired operation and then click OK to save the settings Some configuration settings do not take effect until the system is restarted After completing sections 2 12 1 through 2 12 6 follow the instructions in section 2 12 7 2 12 1 Solvents When you installed the system up to four solvents were connected to the back panel This section of the Instrument Configuration tab allows you to manage these solvents To do S0 The system is shipped with common solvent names already loaded If your solvent names are listed in the s
127. module maximum Line cord is the disconnect device 50 60 Hz 20 to 40 C maximum temperature must be at least 15 C below the boiling point of the solvent Humidity when connected to power 95 relative humidity maximum at 20 to 40 C Flow Rate Range 5 to 200 mL min Flow Rate Accuracy 5 Pressure Limit 2 1379 kPa 200 psi Pressure Accuracy 5 of full scale Gradient Formation Binary gradient from A1 A2 and B1 B2 solvent inlets Solvent selection can be automatically controlled by the method Peak Detection Modes Slope or threshold Flow Cell Pathlength 0 1 mm 25 UV Detection and Monitor Wavelength Selectable from 200 to 360 nm range Optional 200 to 780 nm UV Vis detector Wavelength Accuracy 5 nm Fraction Accuracy 2mL flow rate 60 CombiFlash Rf User Manual Table 1 3 CombiFlash Rf 4x System Specifications Continued Electrical Safety per EN 61010 1 Pollution Degree 2 Installation Category Maximum Altitude 2000 meters Note 1 All specifications are subject to change Note 2 For columns undetected by RFID the max pressure is limited to 50 psi Table 1 4 Component Materials List Component Materials Teledyne Isco RediSep Rf Cartridge Virgin Polypropylene silica based media polyethylene HDPE frits Process Tubing Carbon impregnated fluoropolymer tubing
128. n the instrument s technical reference manual The lightning flash and arrowhead within the triangle is a warning sign alerting you of dangerous voltage inside the product The pinch point symbol warns you that your fingers or hands will be seriously injured if you place them between the moving parts of the mechanism near these symbols gt gt Symboles de s curit Ce symbole signale l existence d instructions importantes relatives au produit dans ce manuel Ce symbole signale la pr sence d un danger d lectrocution Risque de pincement Ces symboles vous avertit que les mains ou les doigts seront bless s s rieusement si vous les mettez entre les l ments en mouvement du m canisme pr s de ces symboles gt gt Warnungen und Vorsichtshinweise Das Ausrufezeichen in Dreieck ist ein Warnzeichen das Sie darauf aufmerksam macht daB wichtige Anleitungen zu diesem Handbuch geh ren Der gepfeilte Blitz im Dreieck ist ein Warnzeichen das Sei vor gef hrlichen Spannungen im Inneren des Produkts warnt Vorsicht Quetschgefahr Dieses Symbol warnt vor einer unmittelbar drohenden Verletzungsgefahr f r Finger und H nde wenn diese zwischen die beweglichen Teile des gekennzeichneten Ger tes geraten gt gt 1 21 CombiFlash Rf User Manual Table 1 5 Hazard Symbols Continued Advertencias y Precauciones Esta se al le advierte sobre la importancia
129. nal computer the summary can be printed on any installed printer If you are attempting to print from the systems touch panel display you must first set up NETWORK PRINTING in the NETWORK CONFIGURATION 6 3 10 2 settings Save As PDF Click this button to save the run summary in PDF file format When viewing the run from a remote personal computer the summary can be saved to any connected storage device If you are attempting to save a PDF file from the touch panel display insert a USB Flash drive in the USB port below the display panel Save As TXT Click this button to save the run summary in an ASCII text file format When viewing the run from a remote personal computer the summary can be saved to any connected storage device If you are attempting to save a TXT file from the touch panel display insert a USB Flash drive in the USB port below the display panel Zoom In Click this button to enable the zoom feature Then click and drag across the region to be magnified Zoom Out Click this button to restore the chromatogram to the normal view Set Scale Click this button to open a window from which you can set the left and right Y axis scales These scales are controlled by the Absorbance and B upper limits Extract Method button Click the Extract button to load a new method file based on the parameters for the run you are viewing Close button Click the Close button when you are do
130. ndows enter a start time and an end time All times are relative to start of the run just following the sample injection Then choose a peak collection mode If you select Peaks the module will collect fluid only when the time window is active and a peak is present If you select All the module will collect all fluid during the time window regardless of the peak state Define up to three time windows 6 38 Section 6 User Help Reference Air Purge You can program the system to purge the column with air after a run This feature allows you to remove solvents from the column before it is discarded Enter the desired time to purge the column in minutes To disable the post run air purge enter zero minutes Peak Collection There are three peak collection options All Peaks or None The All option will collect peak and non peak fluid in tubes The Peaks option will collect only peak fluid in tubes and divert all other fluids to the waste port The None option will divert all fluids peak and non peak to the waste port The None option is useful for developing custom methods that perform a column wash system cleaning or similar function where solvents and elute do not need to be collected When using the A or Peaks options you can specify the maximum Peak and Non peak volumes to be collected in the tubes Be sure to enter volumes less than the maximum tube capacity Different peak and non peak tube volumes can be used to conserve t
131. ne Year Limited Warranty Factory Service for Teledyne Isco CombiFlash Systems This warranty exclusively covers Teledyne Isco CombiFlash instruments providing a one year limited warranty covering parts and labor Any instrument that fails during the warranty period due to faulty parts or workmanship will be repaired at the factory at no e to the customer Teledyne Isco s exclusive liability is limited to repair or replacement of defective instruments Teledy ne Isco is not liable for consequential damages CombiFlash systems are designed to use Teledyne Isco RediSep columns RediSep columns lead the industry in quality and performance Customers may use other brands of columns on their CombiFlash instruments These non RediSep columns may fail to separate compounds may leak silica into the flow path of the CombiFlash instrument and may leak solvents from their fittings and seals Damage to CombiFlash instruments resulting from the failure of a non RediSep column may cause the need for repairs that incur billable service activities The customer is responsible for these charges Teledyne Isco will pay surface transportation charges both ways within the 48 contiguous United States if the instrument proves to be defective within 30 days of shipment Throughout the remainder of the warranty period the customer will pay to return the instrument to Teledyne Isco and Teledyne Isco will pay surface transportation to return the repa
132. ne viewing the run 6 48 Section 6 User Help Reference 6 4 9 1 General instructions for reading the data Most elements on the window are color keyed to help you locate the tubes containing the peaks of interest Below each collected peak there is a color bar that matches a tube in the map on the left The tube map provides a visual representation of the tubes that contain the peaks of interest If you wish to identify the tube by number refer to the table below the rack diagram If the run used multiple racks the Previous and Next Rack buttons are active Click these buttons to scroll through the available racks The number of the currently displayed rack is shown at the top left of the window 6 4 9 2 Reading the Chromatogram The plot area displays the following The red absorbance trace produced by the system s peak detector Absorbance units also shown in red that correspond to this trace are shown on the left Y axis Purple absorbance and green ratio traces may be visible if you are monitoring a second wavelength See the MONITORING THE WAVELENGTH RATIO 6 4 8 help topic The blue gradient curve that was used during the run The Solvent B scale is shown in blue on the right Y axis The X axis depicts the run time shown as minutes or COLUMN VOLUMES ON LINE Vertical lines appear at intervals along the X axis These lines indicate collection tube changes To prevent the plot area from being obscured b
133. nect the threaded fitting on the waste line assembly to the Column Waste port labeled 1 b At the Rf 4x back panel connect the natural tubing of the waste line assembly to the Waste Level Sense port labeled 1 c At the CombiFlash Rf system back panel the black tubing should still be connected to the Inject Valve Waste port from step 1 If not reconnect the tubing d At the waste container cap for channel 1 insert the black tubing into the Diverter Valve Waste opening e At the waste container cap for channel 1 insert the natural tubing into the Waste Level Sense opening Figure 4 5 Push the stop ferrule on the Waste Level Sense tubing against the cap This ensures that the Waste Level Sense tubing is far enough into the container to detect when it is nearly full f At the waste container cap for channel 1 insert the black tubing from the Inject Valve Waste port The completed waste connections for Channel 1 are shown in Figure 4 6 6 Using one of the three remaining waste line assemblies and waste caps complete the waste connections for channel 2 as shown in Figure 4 7 7 Using the three remaining waste line assemblies and waste bottle caps complete the waste connections for channels 2 through 4 as shown in Figure 4 7 4 8 Section 4 Preparation Optional 4x Module All other connections not shown To Waste Bottle 2 Figure 4 7 Channels 2 through 4 waste tubing connections
134. ng the FILE gt OPEN 6 2 1 2 command If you click this option the system reads the RFID tag on the RediSep Rf column loads the previous method and adds a run extension You can then modify the method and click Play to continue the run skipping the equilibration and injection This feature can be useful if the run was interrupted or if additional compounds of interest might be left on the column 6 24 Section 6 User Help Reference Load Previous Run from Detected Rack Rf200 Rf200i and Rf4X This option appears when using the FILE OPEN 6 2 1 2 command If you click this option the system reads the RFID tag on a single rack and displays the RUN FILE 6 1 3 This feature is useful when there is a need to recall run results at a later time Cancel Click the Cancel button to close the window without saving or opening the file 6 3 7 Set Data Path The FILE gt SET DATA PATH 6 2 1 4 menu command opens the Set Data Path window Use this window to select a default folder for the current user After selecting a folder file operations such as saving or opening files will use this selected directory The Set Data Path primarily is used with the USER MANAGEMENT 6 3 12 feature User Management will automatically create a folder for each user When using the system set the data path to your folder or a subfolder within Each user has a different data path The window contains the following controls Current
135. nto AC power Contact Teledyne Isco or its authorized representative for advice Compare the contents of the box with the enclosed packing slips If there are any shortages contact Teledyne Isco immediately The fraction collector arm was stowed to prevent damage during shipping The arm stowing bracket should be removed at this time 1 Loosen the bracket s two thumbscrews along the sides of the arm Figure 3 1 or Do Thumbscrew T Thumbscrew Figure 3 1 Remove arm stowing bracket and spacer 2 Push the arm to the left or right remove the spacer 3 Store the bracket and spacer These parts should be reinstalled if the system needs to be shipped again 3 2 Section 3 Preparation Rf 150 Systems 3 2 Instrument Location The CombiFlash Rf 150 has a relatively small footprint requiring about 1550 square centimeters 240 in of level bench space Ensure that the Combiflash Rf has at least 3 cm 1 25 of air space behind it for ventilation Additional space may be required for solvent and waste containers Refer to Table 1 1 of this guide for environmental conditions and power requirements A WARNING The system is heavy Use a two person lift to prevent injury Before making any connections to the Combiflash Rf 150 place the system on the bench or in the fume hood where it will be operated Temporarily position the system so you can access the back panels to complete the connections
136. nual 2 24 CombiFlash Rf User Manual Section 3 Preparation Rf 150 Systems This section provides instructions for unpacking and installing the CombiFlash Rf 150 system To prepare the system for operation sequentially follow all instructions in sections 3 1 through 3 11 M Note Section 3 13 contains an Installation Qualification checklist If required sign off the checklist entries as you successfully complete the following sections M Note The Rf150 is rated for operation with reagent grade solvents Operation at pressures gt 5 bar 75 psi with high purity hexanes gt 99 pure at 100 concentration may cause the pump to stall and produce a grating sound Many times normal operation can be restored by removing the high pressure load and flushing the pump with isopropyl alcohol 3 1 Unpacking the Unit The Combiflash Rf 150 system is shipped in a single carton Carefully unpack the shipment and inspect the contents Z WARNING The system is heavy Use a two person lift to prevent injury NCAUTION Do not lift the system by the fraction collector arm Use the lifting handles located on the instrument side panels CombiFlash Rf User Manual If there is any damage to the shipping carton or any components contact the shipping agent and Teledyne Isco or its authorized representative immediately A WARNING If there is any evidence that the system has been damaged in shipping do not plug it i
137. ollowing steps a Place the injection syringe on the injection port b Slowly force the liquid into the injection port c Chase the first injection with a second injection solution of at least 0 25 mL The chase solution can be the starting mobile phase or diluted sample mixture d Click OK to continue the run Rf 75 and Rf 150 systems If you selected Equilibration ensure that the solvent delivery tubing is connected to the injection port above the column When you click the OK button the system equilibrates the column and then waits When instructed perform a liquid injection as described in steps a through d above or attach the solid sample cartridge to the injection port and connect the solvent delivery tubing Click the OK button to continue If you selected No Equilibration ensure that the solvent delivery tubing is connected The system starts the run immediately and assumes that the sample has been injected on the column or the solid load cartridge is in place You may allow system to proceed with the run while monitoring the progress on the PeakTrak Main window If desired gradient parameters may be modified during the run See REAL TIME GRADIENT EDITING 6 4 6 6 36 Section 6 User Help Reference As the run progresses the absorbance trace is drawn on the Gradient Plot area The purification continues until the end of the run defined by the run length setting See Also REDISEP RF COLUMN SELECTION GUIDE
138. olumn If the column is reusable the system will also write information to the RFID tag that can help you manage its use The Number of times used First used on and Last used on information can help you determine when the column should be replaced The Last fluid used will help you determine if any solvent remaining in the column will be miscible with the solvent currently used in the system To view the Column Data window open the METHOD EDITOR 6 3 2 window then click the Column Data button 6 3 5 Batch The Batch tab contains the controls to stage and run sequential separations on systems with a 4x Module 6 3 5 1 Queue The Queue section of this tab lets you enable column positions and specify operating parameters Queue Check the box by the column positions that are to be run Note that the Rf 4x system performs the separations sequentially When starting a batch run the system starts with position 1 then continues in numeric order through all selected column positions Sample Name The sample name is a text entry box in which you can label the run Type a unique descriptor 6 22 Section 6 User Help Reference PeakTrak saves the run information under this name Therefore PeakTrak cannot accept any characters that are reserved for the operating system V etc If you do not type a Sample Name PeakTrak will generate a date time stamp for the name when you begin the run
139. olumn mount for Rf 150 systems and insert the column into the top column mount Note that the column fittings are keyed to ensure the correct flow direction 3 Slowly lower the injection valve while aligning the bottom column fitting The spring loaded injection valve will hold the column in place CombiFlash Rf User Manual 4 To seal the column fittings give the column a slight twist 14 turn After loading a RediSep Rf column the system with the exception of the Rf 150 will use RFID technology to automatically detect the media type and column size PeakTrak displays the detected column size on the Main and Method Editor windows If the system does not detect the column manually select the column media and size 5 3 Start a Default Method After completing the system installation steps Section 2 through 4 as necessary preparing the sample and inserting the column you are ready to perform a run with the default method The system s default methods are factory set with run parameters typically used by chemists Default methods are optimized for the use of RediSep Rf columns The default settings will collect all fluid in the fraction collector rack differentiate between UV 254 nm vs non UV absorbing fluid and place the fluids into separate tubes cut UV absorbing peaks based on slope detection or level threshold To start the run from the PeakTrak Main window 1 Enter a sample name If you choose n
140. olvent drop down list boxes skip steps 1 through 3 2 15 CombiFlash Rf User Manual 1 Click the Add Solvent button 2 Enter the solvent name and click the OK button 3 Repeat steps 1 and 2 for all solvents 4 For the Solvent A1 drop down list box select the solvent name connected to the A1 Solvent Inlet Port Repeat for Solvents A2 B1 and B2 5 Select the Enable solvent level sensing check box to enable this feature recommended Clear this box to disable this feature The solvent level sensing feature will monitor solvent usage to minimize the risk of running out of solvent during a purification run The system compares the volume required for a purification run with the estimated volume in the solvent container and alerts you when there is not enough 6 If using the solvent level sensing feature enter the Minimum Solvent Level density dependent as a range in centimeters To understand this range consider that dichloromethane is more dense than hexane If the selected range is 2 4 the system triggers an alert when the dichloromethane level falls to about 2 cm above the weighted filter At the same selected range the system would trigger an alert when the hexane falls to about 4 cm above the weighted filter 2 12 2 General Settings CombiFlash Rf Name optional Use this option to name your system The name will appear in operational displays and run summaries This feature is useful when your
141. on The method file will be stored by the CombiFlash Rf system and will be available for future runs 6 4 3 Alternative Ways to Create Method Files Other than editing a method file on the CombiFlash Rf system there are more ways to create method files 6 4 3 1 Importing Method Files from another System If a method file is located on another CombiFlash Rf system you can use the FILE gt EXPORT METHOD 6 2 1 10 menu command to save the method file on an external storage device From the touch screen panel this storage device is a USB Flash Drive connected to the USB port below the display If connected to the system via an Internet browser the storage device may be any device that can be accessed by the local computer To move the method file onto the system insert the USB flash drive into the USB port or connect to the system from the remote computer then use the FILE gt IMPORT METHOD 6 2 1 9 menu command to locate and import the method file 6 4 3 2 Extracting a Method File from a Previous Run You can perform real time editing during a run to fine tune the method After the run you can extract the run parameters as a method file so it can be used on future runs To do so open the Run file and click the Extract button The system will load a new method with identical run parameters You can then 6 42 Section 6 User Help Reference save the method using the FILE gt Save or Save As menu commands 6 4 3 3 Scaling Method
142. on of Council Directive 2004 108 EC The EMC Directive 200286 E C The WE E EDirective 73 23 EEC The Low Voltage Directive Manufacturers Name Teledyne Isco Inc Manufacturer s Address 4700 Superior Lincoln Nebraska 68504 USA Mailing Address P O Box 82531 Lincoln NE 68501 Equipment Type Environment Laboratory Equipment for Light Industrial Commercial Environments Trade Name Model No CombiFlash Rf Year of Issue 2007 Standards to which Conformity is Declared EN 61010 1 2 edition Safety Requirements for Electrical Equipment for Measurement Control and Laboratory Use EN 61326 1 2006 EMC Requirements for Electrical Equipment for Measurement Control and Laboratory Use Standard Description Severity Applied Performance Criteria EN61000 4 2 Electrostatic Discharge Level 2 4kV contact discharge A Level 3 8kV air discharge EN61000 4 3 Radiated RF Immunity 80 MHz to 1000MHz 80 AM at 1kHz A Level 2 3V m EN61000 4 4 Electrical Fast Transient Level 2 1kV on AC lines A EN61000 4 5 Surge on AC Lines Level 2 1kV common mode A Level 2 0 5KV differential mode EN61000 4 6 Conducted RF on AC lines 150 kHz to 80 MHz A Level 1 1V rms 80 modulated EN6100 4 11 Voltage Dips Interruptions 100 drop 10ms A CISPR11 RF Emissions Group 1 Class A Industrial Scientific and EN 55011 Medical Equipment EN61000 3 2 3 3 Harmonic and Flicker We the undersi
143. or Drain 2 Test the top shelf drain by connecting a vacuum or air supply source to the outlet end of the drain tube Then verify the presence of such vacuum or air supply source on the drain Figure 3 5 3 Route the end of both drain tubes to a suitable waste fluid collection container M Note It may be necessary to extend the drain tube If so splice the tubing with user supplied tubing The user supplied tubing should have an inside diameter no smaller than the existing drain tubing and must be compatible with the solvents used by the system Route this extension tubing to the waste collection vessel 3 8 Section 3 Preparation Rf 150 Systems Vacuum or pressurized air A applied to the outlet end of the drain tube must exist at the top shelf drain hole Figure 3 5 Top Shelf Drain 3 7 Position the System After completing the various connections the system can be moved to its operating position Turn the system so that the operator can access all of the front view features and controls Figure 1 5 Use care not to damage the connections tubing and cables while moving the system CAUTION Ensure that the CombiFlash Rf has at least 3 cm 1 25 of air space behind it for ventilation Position the solvent and waste containers as necessary When using higher density solvents such as dichloromethane DCM place the solvent container level with or above the CombiFlash Rf system Placing solvent containers
144. ork connection using a standard CAT5 cable or for a direct connection to a computer using a cross over cable 16 RS 232 Port Serial communication port to interface with expansion modules 17 USB Port For control of peripheral equipment such as the Rf 4x module This rear panel port is also used in service related functions 18 Cooling Fan Cools the internal electronic assemblies CombiFlash Rf User Manual Z WARNING Do not touch the fan 19 Check Valves Externally mounted pump check valves to simplify pump maintenance The check valves are not present on the Rf 150 systems 20 Solvent Level Sensing Outlet Ports uses air or user supplied gas to measure the hydrostatic pressure of the solvent above the weighted tubing outlet The ports are not present on the Rf 150 systems 21 Solvent Inlet Ports Accepts A and B solvents The Rf 150 system does not use the 4 port block Refer to Section 3 4 for more information on Rf 150 solvent inlet ports 22 Optional External Gas Port An inlet for an internal pump which supplies pressurized air or user supplied gas for solvent level sensing waste container full detection and system column purges The external gas should be regulated to 2 5 psig before connecting the source to this port The port is not present on the Rf 150 systems 23 Mains Power Connects the system to AC line voltage 24 Waste Ports From left to right Waste Level Sense The
145. osition the System 0 000 cece KK KK 2 12 2 9 Install Collection Tube Racks 2 12 2 10 External Detector Optional 2 14 2 11 Turn on Power KK KK KK KK 2 15 2 12 Configure the System 00 0 eee 2 15 2 12 1 Solvents ie ss a ca rw ccc cee 2 15 2 12 2 General Settings 00 000 KK 2 16 2 12 3 Set Date Time 0 0 00 000 ee 2 16 2 12 4 Vapor Limit eee eee 2 17 2 12 5 Set Default Tube Volumes 2 17 2 12 6 User Preferences 0000 eee eee 2 18 2 12 7 Restart the system 00055 2 20 2 13 Prime the Solvent Lines 0 2 21 CombiFlash Rf User Manual 14 System Verification RR RR RR KK 2 22 15 Installation Qualification Checklist 2 22 NN 3 Preparation Rf 150 Systems 3 1 Unpacking the Unit 005 3 1 3 2 Instrument Location 0 0 00 RR KK K8 3 3 3 3 Connect Power cece eee 3 3 3 4 Connect Solvent Lines 00000005 3 4 3 5 Connect Waste Lines 00000 K3 3 6 3 6 Connect and Route Drain Lines 3 7 3 7 Position the System 00 RR RR KK 3 9 3 8 Install Collection Tube Racks 3 10 3 9 Turn on Power dae di sl d ye GAS 3 12 3 10 Configure the System 000000 3 12 3 10 1 Solvents 0 ccc eee 3
146. ot to PeakTrak will enter the date and time as the sample name when you start the run 2 Review the PeakTrak Main window settings If you want to change any settings refer to the Editing a Method on line help topic 3 Click the Play button 4 Select the sample Loading Type from the list This Loading Type should support the sample preparation you chose in Section 5 1 Possible sample loading types for the Rf 200 Rf 200i and Rf 4x systems are Solid Pause Select this option if you intend place the sample into a solid sample load cartridge but 5 6 Section 5 Operation have not yet prepared the sample You must also specify the cartridge type and size When you click the run button the system will perform a column equilibration and then wait while you prepare the cartridge After you have placed the cartridge on the system click OK to continue with the run Solid Select this option if you have prepared the sample and placed it into a solid sample load cartridge You must also specify the cartridge type and size The system will proceed automatically until the end of the programmed run length Liquid Select this option if you have prepared a liquid sample and plan to manually inject it into the injection port after column equilibration None on column Select this option if you have preloaded the sample on the column The system will Skip column equilibration so that the sample will not be flushe
147. r duration on the curve To close the Gradient Table click the Edit Gradient button See Also DEFINING A GRADIENT 6 4 5 6 3 2 5 Peak Collection Use the Peak Collection buttons to set the collection mode All Click this button to collect all fluids in the fraction collection tubes during a run Peaks Click this button to collect only eluted peaks in the fraction collection tubes during a run None Click this button to divert all fluids during a run to the waste port Tube Volume You can also specify the Tube Volume for collected fluids This volume can be the default maximum volume Max option as entered in the 6 18 Section 6 User Help Reference CONFIGURATION 6 3 10 window or a method specific volume less than the capacity defined by the CONFIGURATION 6 3 10 window Note that the actual fraction size may be less if a newly detected peak causes a tube change or if you click the Next Tube button Peak Type or select the desired volume to be collected in each tube when the system detects a peak Non peak Type or select the desired volume to be collected in each tube when peaks are not detected This setting allows you to conserve tubes without diverting non peak elute to waste This setting is ignored when the peak collection mode is set to Peaks or None 6 3 2 6 Peak Detection This section of the window contains option buttons to enable and disable various peak detection op
148. re making any connections to the CombiFlash Rf 4x module place it on the bench or in the fume hood where it will be operated Temporarily position the system so you can access the back panels to complete the connections 4 3 Connect Power Ensure that the On Standby switch below the CombiFlash Rf system touch screen panel is in the Standby position Then use the supplied IEC power cord to connect the CombiFlash Rf 4x module to mains power N WARNING Mains power must meet the voltage frequency and amperage requirements listed on the serial number label N WARNING As long as the AC mains power cord is connected power is inside the unit The mains power cord is the disconnect device Position the CombiFlash Rf system and Rf 4x module so that the power cord can be unplugged or use a power strip where the plug can quickly be removed from the outlet in the event of an emergency 4 2 Section 4 Preparation Optional 4x Module 4 4 Connect Solvent Lines WARNING Risk of fire ignited by electrostatic discharge Never substitute the black tubing on CombiF ash systems The black tubing P N 023 0503 06 is conductive This tubing is required to dissipate static electricity NCAUTION To prevent damage or premature wear to the pump and internal valves clean solvent should be used The solvent should not contain any dissolved solids The CombiFlash Rf system has four solvent ports on the left side panel To complete the
149. resenting a safe setting but one with a reduced margin for error 6 51 CombiFlash Rf User Manual 6 52 CombiFlash Rf User Manual Section 7 Remote Interfaces This section provides instructions for controlling the Combiflash Rf system through a remote interface The system can be accessed by several personal computers through a corporate network or by a single personal computer PC through a direct connection 7 1 Network Configuration The procedures described in this section will require assistance from your network administrator Contact your Information Technology department before proceeding The system has a factory assigned IP address 192 168 1 51 In many cases this address must be reconfigured for use on a corporate network To reconfigure the IP address 1 From the touch screen panel select the Tools Configuration menu command 2 Select the Network Configuration tab 3 Select Static IP as the Network Type The CombiF ash Rf system also provides limited support for DCHP connections If DHCP is desired select DCHP as the network type and skip steps 4 through 7 The system will display instructions required to complete a DHCP connection 4 Enter the IP Address Netmask and Gateway information provided by your network administrator CombiFlash Rf User Manual Should your network administrator request the MAC address a label on the system s back panel lists the Address You also c
150. ridge Rf 200 Rf 200i and Rf 4x Only Occasionally impurities precipitate in the solid load cartridge as compounds are purified This may cause high back pressures resulting in reduced flow rates and long run times Clicking this button changes the injection valve position which removes the cartridge from the solvent path and relieves the back pressure A CAUTION Bypassing the solid load cartridge too soon during the purification run may leave desired compounds on the solid load cartridge After removing the cartridge from the fluid path these compounds cannot be purified If you need to use this feature perform the following to ensure that you do not leave desired compounds on the cartridge 1 Run a TLC plate prior to starting the run Measure the Rf of the slowest eluting compound of interest The Rf should be between 0 2 and 0 8 the same range used by the Gradient Optimizer 6 3 3 Note the Percent B solvent used to run the TLC plate If a compound displays tailing peaks as streaks on TLC the Rf used corresponds to the bottom of the streak 2 Load the Sample in the Solid Load Cartridge and note the cartridge size Refer to list below to determine the cartridge volume If the cartridge is partially filled with silica use the cartridge volume as if the cartridge were completely filled with silica e 5 gram cartridge 10 mL volume e 25 gram cartridge 45 mL volume e 65 gram cartridge 110 mL volume 3 Run the s
151. rmation under this name Therefore PeakTrak cannot accept any characters that are reserved by the operating system V etc If you do not type a Sample Name PeakTrak will generate a date time stamp for the name when you click the Play button to begin the run Flow Rate Type or select the desired flow rate for the run When using the Isco default column methods the system sets the flow rate to the optimum flow rate of the column 6 15 CombiFlash Rf User Manual Equilibration Volume Type or select the volume of solvent that is pumped through the column and out the waste port before the sample is introduced Data is not collected while this volume is being pumped Three or more column volumes are recommended for most applications Initial Waste amp Time Windows Click this button to open a window from which you can view and modify these settings For the Initial Waste type or select the volume of eluate that is allowed to pass through the column to the waste port after the sample is injected After this volume has been diverted the system is ready to collect peak or all fluids in the collection tubes Use this feature to clear the internal lines of any elute left from a previous run and to conserve collection tubes by diverting all fluids until the first peak is expected Time Windows can limit the fraction collection to specific time durations of the run To use time windows first enter a start time an
152. roup 1 Class A Industrial Scientific and EN 55011 Medical Equipment EN61000 3 2 3 3 Harmonic and Flicker The undersigned hereby declares that the design of the equipment specified above conforms to the above Directive s and Standards 79 APO A as of September 5 2008 USA Representative William Foster Vice President of Engineering ANA TELEDYNE ISCO 60 5232 505 A Teledyne Technologies Company DECLARATION OF CONFORMITY Y Application of Council Directive Manufacturer s Name Manufacturer s Address Equipment Type Environment Trade Name Model No Year of Issue Standards to which Conformity is Declared 2004 108 EC The EMC Directive 2002 96 EC The WEEE Directive 2006 95 EC The Low Voltage Directive Teledyne Isco 4700 Superior Lincoln Nebraska 68504 USA Mailing Address P O Box 82531 Lincoln NE 68501 Phone 1 402 464 0231 FAX 1 402 465 3799 Laboratory Equipment for Light Industrial Commercial Environments Rf 150 2013 EN 61326 2006 EMC Requirements for Electrical Equipment for Measurement Control and Laboratory Use EN61010 1 2010 Safety Requirements for Electrical Equipment for Measurement Control and Laboratory Use Standard Description Severity Applied Performance Criteria EN61000 4 2 2008 Electrostatic Discharge Level 2 4kV contact discharge A Level 3 8kV air discharge EN6
153. s This delay accounts for the volume held in the tubing between the detector and the drop former sometimes called the delay volume It is important to remember that the chromatogram displays activity in real time while the fraction collector activity lags behind The duration of this delay will vary with the flow rate Rewind This button is active when the system has completed the run or if the run was stopped by clicking the Stop button The Rewind button returns you to the Main window with the current method If you have stopped the run before its programmed run length has elapsed clicking the Rewind button changes the current method to the new shorter run length This modi fied method is ready for the next run or can be saved for future runs Fast Forward Click this button to jump to the next step of a run Once you have started a run the system performs several steps The first step is to deliver solvents using the pro grammed gradient for the entire run length When this step is complete the system will purge the column with air and clean the injection valve Clicking the Fast Forward button will cause the system to skip any remaining time in the current step and advance to the beginning of the next step Unlike the Rewind button clicking the Fast Forward but ton to complete the run will not modify the current method s run length setting CombiFlash Rf User Manual 5 4 2 Bypassing the Solid Load Cart
154. s are password protected to restrict their use to only trained personnel 6 2 4 3 Software Update Choose this command to open a window used to install patch files to update your CombiFlash Rf system software 6 2 4 4 Export Log Files During operation the system writes operating data to a log file Teledyne Isco service personnel can interpret this data to optimize system performance or to troubleshoot difficulties Teledyne Isco recommends that you use the menu command only when advised by a qualified service technician 6 2 4 5 Last Reorder Code This command should only be used by RediSep Rf Column Plan customers 6 2 4 6 About PeakTrak Choose this command to view information about PeakTrak 6 2 4 7 About Isco Choose this command to view contact information for Teledyne Isco 6 3 PeakTrak Windows This section contains descriptions of the windows used for most PeakTrak tasks In this section you will find information on the following e MAIN WINDOW 6 3 1 e METHOD EDITOR 6 3 2 6 9 CombiFlash Rf User Manual GRADIENT OPTIMIZER 6 3 3 COLUMN DATA 6 3 4 BATCH 6 3 5 FILES 6 3 6 e SET DATA PATH 6 3 7 AUTO PRIME VALVE WASH 6 3 8 MANUAL CONTROL 6 3 9 CONFIGURATION 6 3 10 e SET PASSWORD 6 3 11 USER MANAGEMENT 6 3 12 6 3 1 Main Window The Main window gives you control of the frequently used functions before and during the run After a run or when viewing previous runs
155. s been pumped Solvent A and B selections Select the desired solvents from the list The solvent choices are those defined by the CONFIGURATION 6 3 10 window Rf 200 Rf 200i and 4x only Play button Click this button to start the Auto Prime Fast Forward button Click this button to skip to the next Auto Prime step Cancel button Click this button to stop the Auto Prime and close the window 6 3 9 Manual Control Manual Control can assist with method development maintenance of the system plumbing and system troubleshooting The Manual Control window can be opened by selecting TooLS MANUAL CONTROL 6 2 3 2 from the PeakTrak menu The Manual Control Window has several controls e Channel Select the channel for manual control Rf 4x systems only Solvent A and B Selection Rf200 Rf200i and RfAX Select the desired solvents from the list The solvent choices are those defined by the CONFIGURATION 6 3 10 window Percentage Solvent B type or select the mixture percentage Rf 75 150 systems fixed at 50 6 26 Section 6 User Help Reference Flow Rate Type or select the pump s flow rate in mL min Pump into Tube Rf200 Rf200i and Rf4X As a default setting solvent is pumped to the waste port during manual control If you would like to collect the solvent in a tube select the option button and enter the tube number This function cannot be used with the Was
156. s can often be diagnosed and corrected over the phone or by e mail without returning the instrument to the factory Instruments needing factory repair should be packed carefully preferably in the original carton and shipped to the attention of the service department Small non fragile items can be sent by insured parcel post PLEASE BE SURE TO ENCLOSE A NOTE EXPLAINING THE DEFECT Shipping Address Teledyne Isco Attention Repair Service 4700 Superior Street Lincoln NE 68504 USA Teledyne Isco PO Box 82531 Lincoln NE 68501 USA Repair service 800 775 2965 lab instruments 800 228 4373 samplers amp flow meters Sales amp General Information 800 228 4373 USA amp Canada Fax 402 465 3001 Email Mailing address Phone iscoservice teledyne com Web site www isco com 2g TELEDYNE ISCO Everywhereyoulook February 28 2012 P N 60 2132 281 Rev D
157. s in the 6 23 CombiFlash Rf User Manual current folder the system will ignore the Paste command to prevent the original file from being overwritten Delete Click this button to delete a highlighted file or a folder and its contents New Folder Click this button to create and name a new folder Up Click this button to browse the contents of the next higher folder level Search Enter a keyword and click the Search button to find matching file names Click the Clear Search button to clear the results File Management options Click the Copy Files to Flash Drive or Move Files to Flash Drive to create archive copies PDF or Text of the files on a connected flash drive Click the Delete by Age to specify an age limit beyond which will be deleted from the internal hard drive File folder info scroll box This box lists the contents of the current folder The contents can be sorted by clicking the column headings File Type This option is shown when you can limit the display to certain types of files Select the desired file type from the list File Name This text entry box is used to identify the currently selected highlighted file or folder when browsing and opening files When using a Save As command use this box to name the file Open Save This button performs the listed action Continue Last Run on Detected Column Rf200 Rf200i and Rf4X This option appears when usi
158. s the run length While in the Paused state you can resume the run by clicking the Play button or stop the run by clicking the Stop button If you resume the run the system continues the gradient curve from the B when the system was paused Stop This button suspends the entire run Unlike the Paused state the pump peak detection and fraction collection will not operate While stopped you can save the run data by clicking the Save button abort the run by clicking the Rewind button or resume the run by clicking the Play button Next Tube This button advances the fraction collector to the next tube position This allows you to conveniently collect elute of interest in new tube Rewind This button is active when the system has completed the run or if the run was Stopped by clicking 6 12 Section 6 User Help Reference the stop button The Rewind button returns you to the MAIN WINDOW 6 3 1 If you have stopped the run before its programmed run length has elapsed the current method is modified using the new run length This modified method is ready for the next run or can be saved for future runs Fast Forward Click this button to jump to the next step of a run Once you ve started a run the system performs several steps The first step is to deliver solvents using the programmed gradient for the entire run length When this step is complete the system will purge the column with air Following this th
159. se care not to damage the connections tubing and cables while moving the system 1 Turn the CombiFlash Rf system so that the operator can access all of the front view features and controls Figure 1 5 2 Position the Rf 4x module along the left side of the CombiFlash Rf system 3 The Rf 4x module has a tab extending from the right side of its base plate Slightly raise the Combiflash Rf system and slide the Rf 4x module so that the system will rest on the module s tab The weight of the CombiFlash Rf system on the tab will hold the two together 4 8 Remove Shipping Hardware To prevent damage to the injection valve assemblies during shipping each is secured by a thumbscrew which must be removed before operating the system Firmly grasp the cartridge guide above the injection valve and then remove the thumbscrew Figure 4 9 Gently lower the injection valve once the thumbscrew is removed Save the thumbscrews should there be a need again to stow the valves for shipping CombiFlash Rf User Manual Thumbscrew gt lt one of three E Figure 4 9 Remove thumbscrews 4 9 Turn on Power Turn the CombiF ash Rf system switch to the ON position The system will begin its startup routine which includes self diagnostics During the startup routine the system will detect the connected Rf 4x module and automatically configure the user interface to access the additional channels The system is ready for oper
160. sh Rf User Manual Figure 1 3 CombiFlash Rf 200i system with evaporative light scattering and UV or UV vis detection CombiFlash Rf 200i This system has the same high perfor mance features as the Rf 200 system but includes an inter nal evaporative light scattering detector ELSD During operation this detector can be combined with the UV 200 360 nm or UV vis 200 780 nm detection to isolate visible and UV absorbing compounds as well as compounds with little or no chromophores Section 1 Introduction Figure 1 4 CombiFlash Rf 4x system CombiFlash Rf 4x To perform multiple sequential purifi cations in a personal or workgroup setting a 4x module can be added to the Rf 200 or Rf 200i system This configuration is known as the CombiFlash Rf 4x system a fully auto mated four channel Flash chromatography system The four channels are independent including separate waste containers with full bottle detection All Combiflash Rf systems are optimized for use with Teledyne Isco s RediSep9 Rf columns which are pre packed with a variety of media For example the CombiFlash Rf can 1 5 CombiFlash Rf User Manual purify samples from 4 milligrams to more than 33 grams using RediSep Rf columns loaded with 4 to 330 grams of silica gel Applications include purification of organic compounds for drug discovery as well as research in agrochemicals petrochemicals natural products polymers and catalysts
161. sh to edit and save the system s default methods use the TOOLS gt EDIT DEFAULTS 6 2 3 4 command instead 6 2 1 6 Save Method As Choose this command to open the FILES 6 3 6 window From this window you can rename the current method and save it on the system s internal hard drive The Save Method As command also can scale a method to a different column size To scale a method select the scale option and select the new column size from the drop down list Then click the Save button The system saves a new method file with the gradient and run length scaled to the selected column size 6 2 1 7 Save Run As PDF Choose this command to save the displayed run on a remote computer s hard drive or a USB Flash storage device connected to the system When selecting this menu command from the touch screen the system saves the file on the USB storage device The file name will be the same as the Run file except with a PDF file name extension When selecting this command from a remotely connected computer via a web browser PeakTrak displays a file download window so you can select a location and enter a file name You can open and print PDF files with Adobe Reader 6 2 1 8 Save Run As TXT Choose this command to save the displayed run on a remote computer s hard drive or a USB Flash storage device connected to the system When selecting this menu command from the touch screen the system saves the file on the USB drive
162. size label visible Figure 2 7 insert the racks into the system Slide the rack in until you feel it drop into its seated position Tube Size Label Figure 2 7 Inserting the collection tube rack CAUTION An incorrectly installed rack will cause the rack to be misaligned under the fraction collector arm Misaligned racks might cause fractions to miss the tube opening or deposit in the wrong tube Always ensure the tube size label is visible that is facing outward and the rack is pushed in until it is seated When you turn the power switch to on the system will automatically detect the type of rack and configure program settings accordingly Later in section 2 12 5 you can configure the maximum volume for each tube size 2 10 External Detector Optional CombiFlash Rf systems with serial numbers greater than 209C20092 can be configured for use with an analog signal from a user supplied external detector The analog signal should range from 1 2 to 1 2 volts although custom interface cables may convert other analog signals to this range Contact the factory for interface and fluid path options The external detector option is not supported on CombiF ash Hf 200i systems Section 2 Preparation Rf 200 and Rf 200i Systems 2 11 Turn on Power The system s power switch is located just below the touch screen panel 1 Turn the switch to the ON position The system will begin its startup routine which inc
163. t inlet block were stowed for shipping Refer to the diagram on the back of the system and connect the check valve tubing to the respective ports Finger tighten the fittings 2 Connect the solvent source to the system s Solvent Inlet ports Figure 2 2 shows the recommended connections when the user supplied solvent container has a 38 or 45 mm opening GL 38 or GL 45 If the container does not have a GL38 or GL45 opening do not use the cap Use an alternate method to cover the open ing to prevent particulate matter from entering the con tainer Ensure that the solvent and air tubing are secured so that the weighted filter remains at the bottom of the sol vent source The Solvent Level Sensing air line connects to the back panel with a compression fitting This fitting must be air tight To connect tubing using the compression fittings 3 Slide the nut and ferrule onto the tubing as shown in Figure 2 2 4 Fully insert the tubing into the threaded bulkhead fitting on the back panel of the system 2 5 CombiFlash Rf User Manual 5 Finger tighten the nut onto the threaded bulkhead fitting This will seat the ferrule in the fitting M Note When using higher density solvents such as dichloromethane DCM place the solvent container level with or above the CombiFlash Rf system Placing solvent containers below the level of the system can contribute to decreased flow due to the high vapor pressure of DCM This problem b
164. t 1kHz A Immunity Level 2 3V m EN 61000 4 4 Electrical Fast Level 2 1kV on AC Lines A Transient EN 61000 4 5 Surge on AC Lines Level 2 1kV common mode A Level 2 0 5kV differential mode EN 61000 4 6 Conducted RF on AC 150 kHz to 80 MHz A Lines Level 1 1V rms 80 modulated EN 61000 4 11 Voltage 0 during half cycle and full cycle A Dips Interruptions 70 at 25 cycles CISPR11 EN 55011 RF Emissions Group 1 Class A Industrial Scientific and N A 2009 A1 2010 Medical Equipment EN 61000 3 2 3 3 Harmonic Flicker N A N A The undersigned hereby declares that the design of the equipment specified above conforms to the above Directive s and Standards as of April 11 2012 USA Representative p A O A William Foster Director of Engineering Part Number 605232253 Rev A ANA TELEDYNE ISCO A Teledyne Technologies Company END 04 005 8 29 08 DECLARATION OF CONFORMITY ISMEA Application of Council Directive Manufacturer s Name Manufacturer s Address Equipment Type Environment Trade Name Model No Year of Issue Standards to which Conformity is Declared 2004 108 EC The EMC Directive 2006 95 EC The Low Voltage Directive 2002 96 EC The WEEE Directive Teledyne Isco Inc 4700 Superior Lincoln Nebraska 68504 1398 USA P O Box 82531 Lincoln NE 68501 2531 Phone 1 402 464 0231 Facsimile 1 402
165. t the ELSD option is selected Otherwise the ELSD may be disabled The Rf 200i system will automatically check this option if the previous run used the ELSD 8 Click OK and the run begins The system responds according to the sample Loading Type selected in step 4 Rf 200 Rf 200i and Rf 4x systems If you selected Solid Pause the system equilibrates the column with the starting 6B and then waits while you prepare the solid sample load cartridge Place the cartridge on the system and click OK to continue e If you selected Solid the system equilibrates the column and immediately starts the separation The prepared solid load cartridge should be in place before clicking the OK button e f you selected None on Column the system skips the equilibration and immediately starts the separation The sample must be on the column before clicking the OK button If you selected Liquid the system equilibrates the column and then waits When instructed perform the following steps a Place the injection syringe on the injection port b Slowly force the liquid into the injection port c Chase the first injection with a second injection solution of at least 0 25 mL The chase solution can be the starting mobile phase or diluted sample mixture d Click OK to continue the run 5 8 Section 5 Operation Rf 150 systems e Ifyou selected Equilibration ensure that the solvent delivery tubing is connected to the inj
166. te Flow Path valve position This feature can be useful when trying to recover a compound that has precipitated or crashed somewhere in the fluid path e Valve Position Select the valve position for the desired solvent flow path Rf 200 Rf 200i and Rf 4x systems only Prime A Click the Prime A button to pump 100 of solvent A at the selected flow rate Prime B Click the Prime B button to pump 100 of solvent B at the selected flow rate Prime X B Click this button to pump a mix of Solvent A and B according to the Percentage Solvent B value Rf 75 and Rf 150 systems fixed at 50 Stop Click this button to stop the pump This button is only active while the pump is running The window also includes controls to air purge Rf 200 Rf 200i and Rf 4x systems Air Purge Cartridge Click this button to pump compressed air or external gas through the solid sample cartridge Air Purge Column Click this button to pump compressed air or external gas through the column Stop Click this button to stop the air purge This button is only active while purging The right half of the window displays the raw lamp energy Click the Close Manual Control button to close the window See Also MANUAL CONTROL OF THE COMBIFLASH RF 6 4 10 6 27 CombiFlash Rf User Manual 6 3 10 Configuration The Configuration window allows you to define the operating characteristics of your CombiF lash
167. te locations 7 2 Direct Connection A direct connection supports communication between the CombiFlash Rf system and a single PC that meets the recommendations in Table 7 1 This guide provides instructions for Windows 7 7 2 1 Windows Vista 7 2 2 Windows XP 7 2 3 and Windows 2000 section 7 2 4 Refer to the applicable section then follow the instructions in section 7 2 5 to complete the direct connection A direct connection requires a crossover CAT5 cable Standard CAT5 network cables will not work You can purchase a crossover cable locally or ask your Information Technology department to assemble one You can also order a crossover cable part number 480 6545 02 from Teledyne Isco M Note These instructions assume that the system is using the factory set IP address of 192 168 1 51 7 4 Section 7 Remote Interfaces The following instructions may require a user account with Administrator privileges on the PC If you cannot modify the settings in sections 7 2 1 through 7 2 4 contact your Information Technology department 7 2 1 Windows 7 Settings 1 From the Windows Start menu select Control Panel then select the Network and Sharing Center option The PC displays the Network and Sharing Center 2 Select the Change Adapter Settings option to display the available network connections 3 Right click on the Local Area Connection icon and select the Properties option 4 Select Internet Protocol Version
168. the checklist entries as you successfully complete the following sections M Note The optional 4x module is not compatible with the CombiFlash Rf 75 system 4 1 Unpacking the Unit The CombiFlash Rf 4x module is shipped in a single carton Carefully unpack the shipment and inspect the contents If there is any damage to the shipping carton or any components contact the shipping agent and Teledyne Isco or its authorized representative immediately A WARNING If there is any evidence that the system has been damaged in shipping do not plug it into AC power Contact Teledyne Isco or its authorized representative for advice Compare the contents of the boxes with the enclosed packing slips If there are any shortages contact Teledyne Isco immediately 4 1 CombiFlash Rf User Manual 4 2 Instrument Location The CombiFlash Rf system has a relatively small footprint requiring about 1550 square centimeters 240 in of level bench space Ensure that the CombiFlash Rf has at least 3 cm 1 25 of air space behind it for ventilation Additional space may be required for solvent and waste containers The optional 4x expansion module is installed to the left of the CombiFlash Rf system This module requires an additional 933 cm 145 in of level bench space Ensure that the module has at least 3 cm 1 25 of air space behind it for ventilation Refer to Table 1 3 for environmental conditions and power requirements Befo
169. the selected range is 1 2 an alert would be triggered when the dichloromethane falls to about 1 cm above the bottom of the weighted filter At the same selected range an alert would be triggered when the 6 28 Section 6 User Help Reference hexane solvent level falls to about 2 cm above the weighted filter CombiFlash Rf Name Enter a name for the system This name will appear in the title bar of the Main window The name will also appear in run summaries Unique names are helpful when operating more than one system Time Zone Select your local time zone Set Date Time Click this button to set the date and time From the touch screen this command opens a window that allows you to set the instrument date and time If you select this command from a remotely connected computer this menu command automatically sets the system time to match the computer s Note The system will restart after you set the time To prevent unauthorized time changes this feature is password protected See also SET PASSWORD 6 3 11 Vapor Limit Type or select the vapor limit setting The CombiFlash Rf system s internal vapor sensor detects vapors present in the immediate atmosphere When this value is exceeded the system will shut down to avoid a hazardous condition For more information refer to SETTING THE VAPOR LIMIT 6 4 11 This function is an optional addition to the Rf150 Set Default Tube Volume Click this button to open a window from w
170. tion of the peak the compound eluting may not be pure To display the ratio open the METHOD EDITOR 6 3 2 window and select the Show Ratio option 6 4 9 Viewing runs After completing a run the PeakTrak Main window is used to display all collected run data You can also open previous runs to view the chromatogram and the peak tube locations To open a previous run 1 From the main window select File gt Open or click the Open button The FILES 6 3 6 window is displayed 2 Choose a Run file and click the Open button 3 PeakTrak displays the run in the MAIN WINDOW 6 3 1 The PeakTrak Main window will include Rack and tube information The left pane of the window lists the current rack a map for that rack and a table that lists the peaks and their corresponding tube numbers If the window is currently displaying the collection parameters click the Rack View button to view this information Method parameters Click the Display Method button to view a summary of the peak detection and collection settings for the run Chromatogram The right side of the window displays the chromatogram identified by the Sample Name in the title bar of the window 6 47 CombiFlash Rf User Manual Option buttons The option buttons give you quick access to frequently used commands while viewing a run The buttons are e Print Click this button to print a run summary When viewing the run from a remote perso
171. tions all other connections not shown 4 6 Section 4 Preparation Optional 4x Module To Rf 4x Module To Hf 4x module Waste Level Sense Column Waste connections connections 1 2 3 or i 1 2 3 0r4 Channel 1 Only To Hf system Inject Valve Waste 60 0923 017 Ferrule 60 0923 015 Compression Fitting See step 8 for details on Nu this connection for channels 2 through 4 60 5234 188 Waste Line Assembly Ventilation Hole Luer fitting If necessary connect user supplied tubing and route to fume hood See Top View Tubing Stop ferrule 60 5235 053 38 mm Waste Cap or 60 5235 054 45 mm Waste Cap Top View Column Waste Waste Level Sense with tubing stop O eC Ventilation Inject Valve Waste Rf 200 or Valve Waste Out Rf 4x Figure 4 5 Recommended Rf 4x module channel 1 waste connections 4 7 CombiFlash Rf User Manual 5 Select one of the four waste line assemblies PN 60 5234 188 from the accessory package This will be used to complete the waste connections for channel 1 as shown in Figure 4 6 These instructions assume that the column waste from each channel will be isolated in separate containers If the waste is to be collected in a common container omit the use of the waste caps and secure all liquid and air lines in the common container The ends of all air lines should extend at least 2 inches b cm below the container opening a At the Rf 4x back panel con
172. tions When an option is enabled the window also allows you to modify the setting details for that peak detection option All enabled options will be displayed on the chromatogram The options and settings for the Rf 200 Rf 200i and Rf 4x systems are described below Note You can select up to four peak detection options on the Rf 200 Rf 200i and Rf 4x systems If using more than one option such as A1 with 2 the system considers a peak to be present when any option is true a logical OR l and 22 wavelength 1 and 2 Enable these options to use and configure primary and secondary wavelength detection When enabled type or select the peak detection wavelength in nanometers then click the Details button to configure additional settings described below Slope Based Select this option to enable Slope based peak detection When enabled peaks will be recorded if the slope algorithm indicates a peak within the Peak Width setting Peak Width Select the average peak width setting Peak widths are measured at the baseline The slope detector will detect peak widths ranging from about 0 2 to 2 times the peak width setting For 6 19 CombiFlash Rf User Manual example if you entered a peak width of 1 minute the range would be 12 seconds to 2 minutes For best operation the peak width should be set to just over the average peak width being separated For instance if the average peak width is 45 seconds yo
173. tory practices and standard safety procedures 1 19 CombiFlash Rf User Manual N WARNING The CombiFlash Rf has redundant safety devices to limit pressure to less than 210 psi 1448 kPa RediSep columns smaller than 100 g are CE certified using standard IEC61010 1 for use on the CombiFlash Rf RediSep columns larger than 100 g meet Pressure Vessel Directive 97 23 EC Teledyne Isco strongly recommends against the use of columns rated less than 210 psi 1448 kPa 1 6 1 Hazard Severity Levels This manual applies Hazard Severity Levels to the safety alerts These three levels are described in the sample alerts below A CAUTION Cautions identify a potential hazard which if not avoided may result in minor or moderate injury This category can also warn you of unsafe practices or conditions that may cause property damage A WARNING Warnings identify a potentially hazardous condition which if not avoided could result in death or serious injury N DANGER DANGER limited to the most extreme situations to identify an imminent hazard which if not avoided will result in death or serious injury 1 6 2 Hazard Symbols The equipment and this manual use symbols used to warn of hazards The symbols are explained in Table 1 5 1 20 Section 1 Introduction Table 1 5 Hazard Symbols Warnings and Cautions The exclamation point within the triangle is a warning sign alerting you of important instructions i
174. ts A and B will be mixed gradually from the last programmed B until the next programmed B is reached This method connects the inflection points that define the gradient with a straight line If the Step method is selected each B value is held until the next programmed B is reached Because the B values are held the changes at each new value are nearly instantaneous producing a stepped appearance on the plot area e f Isocratic is selected the system will pump a single B mix throughout the run Enable Rapid Equilibration Select this option to turn on rapid equilibration When enabled this time saving feature pumps solvent at the maximum flow rate during column equilibration Note that some column sizes and types might be adversely affected by high flow rates 6 3 11 Set Password This window opened by selecting the TOOLS gt SET PASSWORD 6 2 3 6 menu command is used to enter and change the system password and to enable password protection for the default methods To enter or change a password 1 Type the password The factory default password is combiflash 2 Type the password again to verify your entry 3 To protect the Default methods select the Require Password For Editing Default Methods option 4 Click the OK button to save your settings and close the window The following menu commands will require a password before displaying the requested window TOOLS gt EDIT DEFAULTS 6 2 3 4 TOOLS gt USER
175. u should enter a peak width of 1 minute For most flash chromatography 1 minute is a good starting point for a peak width Threshold Detection Select this option to enable Threshold peak detection When enabled peaks will be recorded if the Absorbance Units AU value is exceeded Threshold Type or select the Absorbance Units value to be used for Threshold detection Note If both Slope and Threshold peak detection methods are checked the system considers a peak to be present when any one condition is met This logical OR operator means that the system will cut a peak when either the Slope condition is true or when the Threshold condition is true Monitor Enable this option to only use the detection source as a monitor When enabled the detection source will be displayed as a trace on the gradient plot area but will not be used to cut peaks All Wavelength Detection Enable this option to detect peaks within a user selected range of wavelengths When enabled click the Details button to configure additional settings These settings include the slope based and peak width options described in 41 and A2 above Additionally you can type or select the minimum and maximum wavelengths limits in nanometers External Detector Enable this option to use a 0 to 1 volt analog signal from an external detector When enabled click the Details button to configure additional settings These settings include the slope b
176. ubes when using the All peak collection mode by collecting large volumes of non peak fluid while creating more smaller fraction volumes of fluids of interest The fraction collector will advance to the next tube whenever a peak is detected Also note that the system will advance to the next tube if a new peak is detected before completing the last one sometimes called a double advance Keep in mind that the slope and threshold detection methods use algorithms which optimize the peak detection For example peak detection includes a hysteresis to reduce the likelihood of multiple tube advances that may occur if there is noise at the beginning or tail end of a peak Peak Detection This section of the window contains option buttons to enable and disable various peak detection options When an option is enabled the window also allows you to modify the setting details for that peak detection option All enabled options will be displayed on the chromatogram The options and settings for the Rf 200 Rf 200i and Rf 4x systems are described below Note You can select up to four peak detection options excluding the External option on the Rf 200 Rf 200i and Rf 4x systems If using more than one option such as 41 with 2 6 39 CombiFlash Rf User Manual the system considers a peak to be present when either option is true a logical OR l and 42 wavelength 1 and 2 Enable these options to use and configure primary and second
177. ue use of the CombiF ash Rf 200i if liquid is present at the Pump Drain Contact Teledyne Isco technical service for assistance with correcting the leak CombiFlash Rf User Manual Figure 1 9 Optional CombiFlash Rf 4x Module Features Back 32 Waste Level Sense The system uses air or user supplied gas to measure the hydrostatic pressure of the waste above the level sense line end The system will suspend operation when this level is too high Four ports allow you monitor the level of each waste container on the Rf 4x system 33 Column Waste Waste fluids are sent out ports one through four Tubing connected to these ports are paired with the waste level sense air lines and routed to a collection container for each channel 34 From Rf This port on the Column Waste block receives fluid from the diverter valve on the fraction collector arm 35 Valve Waste Out The injection valves on channels 2 through 4 are cleaned after each run The system directs fluids from this automatic cleaning routine to this shared port 36 A B reserved for future use 37 USB Cable This cable connects to the USB port on the Rf 200 or Rf 200i system back panel see item 17 The Rf Section 1 Introduction system controls the operation of the Rf 4x module through this connection 38 Mains Power Connects the module to AC line voltage 39 Waste Level Sense From Rf Receives pressurized air or optional external gas from the
178. uid spray The instrument is not watertight and these actions could damage the internal electronics 8 2 Section 8 Maintenance 8 3 1 Collection Rack and Tray Cleaning WARNING Risk of fire or equipment damage Unclean collection racks and tray might inhibit their conductive properties The racks and tray must be kept clean to dissipate static electricity The collection tube racks and tray are made of conductive plastic Dirt film or coatings might prevent their ability to dissipate static electricity To avoid problems that possibly result from an electrostatic discharge clean the racks and tray monthly Use distilled water with a mild detergent For tougher stains use isopropyl alcohol 8 4 Tubing Inspection A WARNING Risk of fire or equipment damage Faulty tubing fittings and drains may allow organic solvents to pool in unsafe areas creating a potential for dangerous levels of flammable vapors Improper draining may damage the instrument s internal components Perform a tubing inspection monthly 1 Visually inspect the solvent waste and drain tubing The tubing must be free of any damage kinks or deterioration Fittings should show no signs of leaks 2 Test the collection tray drain and top shelf drain by connecting a vacuum or air supply source to the outlet end of the drain tubes Then verify the presence of such vacuum or air supply source on the drain hole Figures 8 1 and 8 2 Correct any
179. urge peak detection and peak collection mode 6 37 CombiFlash Rf User Manual Equilibration Volume Column equilibration is optional but you may find that separations are more effective if the column is equilibrated before injecting the sample The recommended equilibration volume is at least three column volumes of the starting mobile phase RediSep Silica gel column volumes are RediSep Rf 4 g 4 8 mL RediSep Rf 12 g 16 8 mL RediSep Rf 24 g 33 mL RediSep Rf 40 g 48 mL RediSep Rf 80 g 125 mL RediSep Rf 120 g 192 mL RediSep Rf 220 g 334 mL RediSep Rf 330 g 443 mL Other media types will have similar column volumes Refer to the REDISEP RF COLUMN SELECTION GUIDE oN LINE for actual volumes As an example consider a RediSep Rf 4 g column and a 3 column volumes setting Before instructing you to inject the sample the system will pump 14 4 mL of solvent A through the column and out the waste port Initial Waste amp Time Windows The initial waste diverts a user determined line volume to the waste port until the eluant is about to be collected At the start of a run the internal solvent lines hold some solvent from the previous run This volume can be diverted to waste along with the anticipated volume of fluid that will pass through the column before a compound of interest will elute Time Windows can limit the fraction collection to specific time durations of the run To use time wi
180. ve solvent from a prepared cartridge by attaching the cartridge cap and manually purging TOOLS gt MANUAL CONTROL the cartridge with air Empty solid sample cartridges also allow a variety of adsorbents such as diatomaceous earth boiling chips cotton balls or paper wipes Even with alternative adsorbents always use top and bottom frits to prevent fluid path problems 5 2 Section 5 Operation After you have prepared the pre filled or empty cartridge place the solid sample cartridge on the system 1 Attach the desired adjustable cartridge cap e 60 5237 047 fits 2 5 and 5 gram solid load cartridges e 60 5237 048 fits 12 and 20 gram solid load cartridges e 60 5237 044 fits 32 and 65 gram solid load cartridges 2 Press the lever on the side of the cap and fully extend the plunger Figure 5 1 Figure 5 1 Press the lever and extend the plunger 3 Slide the plunger into the cartridge until it reaches the top frit Figure 5 2 4 Press the lever and push the cartridge into the cap Align the cartridge so that it fits fully into the recess in the cap Figure 5 3 5 Load the solid sample cartridge with cap on the sample injection port M Note Earlier Combi Flash Rf solid load cartridge caps required the use of sleeves Contact the factory for information on upgrading these older systems to use the new cartridges without sleeves 5 8 CombiFlash Rf User Manual Figure 5 2 Insert the plunger to the bed
181. w cell cleaning on line help topic To perform a quick cleaning Section 8 Maintenance 1 After clicking the Cancel Run button select the Tools gt Manual Control menu option 2 From the Manual Control window Figure 8 5 note the Raw Lamp Energy level at 254 nm The Raw Lamp Energy gauge has three ranges red yellow and green RAW LAMP ENERGY 1000 at 81 ms The flow cell should be cleaned for optimal detector performance A dirty flow cell may cause the detector signal to appear noisy FLOWY CELL CLEANING HELP Figure 8 5 Raw Lamp Energy Gauge e Red lamp energy is obstructed to a degree that the system might not reliably detect peaks If you attempt to operate the system peak collection will be forced to collect all This prevents diverting desired compounds to waste Yellow lamp energy is partially obstructed and the peak detector performance might be impaired The system can be operated although the absorbance trace might appear noisy Yellow might be a normal indication for systems with flow cell windows stained by compounds Green lamp energy is sufficient to detect peaks up to 2 4 Absorbance Units 3 Remove the column and insert a bypass tube between the upper and lower column mounts A bypass tube P N 209 0165 46 can be found in the accessory kit P N 60 5239 006 4 Set the Flow Rate to 40 mL min Figure 8 6 8 7 CombiFlash Rf User Manual SOLVENT A hexane PRIME A SOL
182. y seated in the port 3 Cut the tubing to length and connect the other end to the user supplied carrier gas An assortment of l s inch adapters are supplied in the accessory kit to complete the connection to your gas source Teledyne Isco recommends gt 99 pure nitrogen from a source that can deliver 2 5 SLPM at 60 to 70 psi 2 7 Connect and Route Drain Lines WARNING Risk of fire or equipment damage Failure to connect drain lines may allow organic solvents to pool in unsafe areas creating a potential for dangerous levels of flammable vapors Improper draining may damage the instrument s internal components The CombiFlash Rf system has drain tubes extending from its back panel The tubes drain away any liquid spilled on the top shelf Rf 200 only and the tray beneath the fraction collection racks 1 Test the Fraction Collector drain by connecting a vacuum or air supply source to the outlet end of the drain tube Then verify the presence of such vacuum or air supply source on the drain Figure 2 4 2 Rf 200 only Test the top shelf drain by connecting a vacuum or air supply source to the outlet end of the drain tube Then verify the presence of such vacuum or air supply source on the drain Figure 2 5 CombiFlash Rf User Manual Vacuum or pressurized air applied to the outlet end of the drain tube must exist at the collection tray drain hole Figure 2 4 Fraction Collector Drain R
183. y tube change marks PeakTrak may limit the number of marks 6 4 10 Manual Control of the CombiFlash Rf Manual Control can assist with method development maintenance of the system plumbing and system troubleshooting To manually control the CombiFlash Rf system first open the MANUAL CONTROL 6 3 9 window by clicking the Tools Manual Control menu command 6 49 CombiFlash Rf User Manual 6 4 10 1 Pumping Solvents To pump either solvent select the solvent from the Solvent A or Solvent B lists Then click the Prime A or Prime B buttons to pump the selected solvent To pump a mixture or Solvent A and B adjust the Percentage Solvent B setting and then click the Prime __ B button When finished click the Stop button You can control the flow rate by adjusting the Flow Rate mL min setting By default the system pumps the solvent directly to the waste port With Rf 200 Rf 200i and Rf 4X systems you can change the fluid path by selecting a different Valve Position option through column or through cartridge and column For valve positions other than Direct To Waste you can pump the solvent into a collection tube To do so select the Pump Into Tube option and then select which tube next or specific tube number 6 4 10 2 Air Purging On CombiFlash Rf 200 Rf 200i and 4x systems you can purge the solid sample cartridge and column path with air To air purge a path click the Air Purge Solid Sample or Air Purge

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