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1260 Infinity Micro-scale Fraction Collector/Spotter User Manual

1. Control of Thermostat Homote CAN DC Out RS232C ll vaen Ke ower cy HOT 7 N q Figure 3 Cable Connections AC Power CAN cable to next module The micro scale fraction collector spotter is turned ON when the line power switch is pressed and the green indicator lamp is illuminated The detector is turned OFF when the line power switch is protruding and the green light is OFF WARNING To disconnect the micro scale fraction collector spotter from the line unplug the power cord The power will supply still uses some power even switch at the front panel is turned OFF Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 23 1 Installing the Micro scale Fraction Collector Spotter Installing a Thermostatted Fraction Collector Preparation Locate bench space Provide power connections Unpack the micro scale fraction collector spotter and the thermostat Parts required Fraction Collector and thermostat Power cords ChemStation or Agilent Instant Pilot G4208A 1 Place the thermostat on the bench 2 Remove the front cover and route the condensation drain tube to a waste con
2. Fraction Trigger Mode C Off Use Timetable Peak based Max Peak Duration 1 0 min p Peak Detectors Detector Working Mode Up Slope Unit s Down Slope Unit s Threshold Unit Upper Threshold Unit Unit 1 DADI Off 5 00 5 00 5 000 3000 000 mAU 2 UIB Off 5 00 5 00 5 000 3000 000 mv Fraction is collected when a peak is detected by F Use MSD for Mass based Fraction Collection all selected peak detectors C at least one selected peak detector _Fraction Preview r Timetable l t Time Trigger Mode Max Peak Duration min of Fractions Timeslices min Contact Adj K 1f oo w amma _ 2 3 1000 Off Copy Paste gt Time Auxiliary Auxiliary Max fill volume per location as Pump j a Stoptime Palen min IV Recovery on the track as configured ul Posttime Off min Contact Control Adjustment 1 0 I Store temperature signal I Store delay sensor signal I Store UIB signal OK Cancel Help Figure 6 Set up Fraction Collector dialog box Agilent 1260 Infinity Micro sc ale Fraction Collector Spotter User Manual 49 2 2 Configuration and Operation of the Micro scale Fraction Collector Spotter CAUTION Fraction Trigger Mode The Fraction Trigger Mode can either be Use Timetable or Peak based can be selected Furthermore for the Peak based mode a time for Maximum Peak Duration can be specified When the Peak based mode is chosen it overrules all settings in the Tim
3. Agilent Technologies Notices Agilent Technologies Inc 2010 2013 No part of this manual may be reproduced in any form or by any means including elec tronic storage and retrieval or translation into a foreign language without prior agree ment and written consent from Agilent Technologies Inc as governed by United States and international copyright laws Microsoft is a U S registered trademark of Microsoft Corporation Manual Part Number 61364 90013 Rev B Edition 10 2013 Printed in Germany Agilent Technologies Hewlett Packard Strasse 8 76337 Waldbronn Warranty The material contained in this docu ment is provided as is and is sub ject to being changed without notice in future editions Further to the max imum extent permitted by applicable law Agilent disclaims all warranties either express or implied with regard to this manual and any information contained herein including but not limited to the implied warranties of merchantability and fitness for a par ticular purpose Agilent shall not be liable for errors or for incidental or consequential damages in connec tion with the furnishing use or per formance of this document or of any information contained herein Should Agilent and the user have a separate written agreement with warranty terms covering the material in this document that conflict with these terms the warranty terms in the sep arate agreement shall cont
4. Figure 7 Fraction preview dialog box The Fraction Preview screen allows testing the fraction collection parameters against a sample chromatogram It can also be used to optimize the fraction collection parameters interactively With the help of this tool values for up and down slope as well as for upper and lower threshold can easily be graphically specified A chromatogram e g a pilot run can be loaded by pushing the Load Signal button Parameters can now be changed either manually in the Detector Table and Timetable or graphically in the Fraction Preview screen By pushing the desired buttons on the right hand side of the Fraction Preview screen the chromatogram can be zoomed the values for up and down slope can be specified and the upper and lower threshold level can be set up The graphically specified values are automatically transferred to the Peak Detector Table Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Configuration and Operation of the Micro scale Fraction Collector Spotter 2 Starting Your Run with Fraction Collection or MALDI Spotting CAUTION Resetting the current fill levels To protect the collected fractions spots for contaminations and to avoid overfilling of single positions the Agilent 1260 Infinity Micro scale Fraction Collector Spotter stores current fill levels on the module and blocks used positions If a tray with new well plate or MALDI target is inserted in the micro sca
5. Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 103 4 Maintenance and Simple Repairs 2 Press the yellow clips of the capillary guiding assembly 3 Slide the new capillary guiding assembly into the holder of together and pull the assembly towards the rear of the the capillary carrier assembly Make sure to push it all the capillary carrier assembly way to the front 4 Reinstall the Capillary the clamp nut and the wellplate On completion of this procedure adapter IMPORTANT Do not tighten the clamp nut without a Re install the tray s in the tray base capillary installed lt a Start the instrument 7 E Close the front cover p clamp nut 104 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Maintenance and Simple Repairs 4 Exchanging the Flap Septum and the Waste Tubing Frequency When defective or contaminated When waste port is clocked Tools required None Parts required Flap Septum G1364 27107 Waste Tubing G1364 86711 Before beginning this procedure 1 Remove the internal tray from the instrument see Exchanging the Internal Tray on page 107 Don t snap Position the transport unit of the micro off or squeeze the waste tubing collector spotter in the Home Position see Deactivating the Door Lock Sensor on page 74 and remove all
6. 1 If the transport unit has been exchanged orifit See Service Manual G1364 90111 for details is strongly misaligned set the 8 bit configuration switch to the Forced Cold Start Configuration 2 Install the 4 well plate tray G1364 84511 or IMPORTANT Remove all plates G1364 84521 3 Ensure that the wellplate adapter is correctly assembled Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 77 3 Troubleshooting and Test Functions How to perform a Transport Unit Self Alignment Steps Comments 4 Select the Maintenance menu in the Diagnosis view of the Agilent ChemStation 5 Inthe menu choose Fraction Collector gt The Transport Alignment Procedure takes Transport Alignment to start the automated approximately 10 15 minutes procedure 6 Set the 8 bit configuration switch to the default See Service Manual G1364 90111 for details setting 78 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Troubleshooting and Test Functions 3 Arm Length Calibration The Arm length calibration is only be required for 384 well plates This procedure calibrates for the arm length of the capillary carrier assembly The arm length calibration is required e after disassembling the transport unit capillary carrier assembly e if problems with the positioning of the capillary tip occur in particular on plate 4 back right We recommend to switch on the lamp during the calibra
7. Adjust the needle in such a way that it touches all corner positions of the target accurately in their center xy position alignment buttons r Position mm Height mm 7 Up eae Current Position Corrective Values mm Al X Axis 0 Y Axis 0 Set To Default Z Axis 0 Cancel Corrective Values relative to the previous alignment Figure 19 Position Accuracy Calibration Interface for well plates Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 83 3 Troubleshooting and Test Functions Position Accuracy Calibration for MALDI Targets The Position Accuracy Calibration for MALDI targets has to be performed if e a different type of MALDI Target is used e the positioning is inaccurate e after mayor repairs e g exchange of transport assembly capillary carrier or mainboard e after a firmware update We recommend to switch on the lamp during the calibration Open the command line in the ChemStation and type microafclampon to switch the lamp ON microafclampoff to switch the lamp OFF During the position accuracy calibration the door lock sensor is disabled The fraction collector arm will move while the front door is open To avoid personal injury keep fingers away from the needle area during position accuracy calibration All users must be informed about this potential risk CAUTION Th
8. Use the left and right button in the dialog box Figure 18 to adjust the capillary tip to the center of the well Click the Plate 3 button in the Arm Length Calibration dialog box Figure 18 on page 81 Use the left and right button to adjust the capillary tip to the center of the well After adjusting the capillary this way the determined values are stored by pressing the OK button Click on Explain to get detailed instructions and information The capillary carrier will move to well position A7 on plate 1 The capillary carrier will move to well position A18 on plate 3 It might take some time to store the positions on the micro scale fraction collector spotter Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Troubleshooting and Test Functions 3 Arm Length Calibration rm Length Calibration Figure 18 Arm Length Calibration Interface Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 81 3 Troubleshooting and Test Functions Position Accuracy Calibration for 384 Well Plates We recommend to perform the Position Accuracy Calibration for 384 well plates e if the positioning is inaccurate e after mayor repairs e g exchange of transport assembly capillary carrier or mainboard e after a firmware update e if a different type of 384 well plate is used We recommend switching on the lamp during the calibration Open the command line in the ChemSt
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10. Figure 3 Positions of the capillary tip Capillary Alignment for MALDI Spotting If the micro scale fraction collector spotter is used for MALDI spotting the well plate adapter has to be replaced with the spotting adapter G1364 83205 which is supplied in the accessory kit see Table 18 on page 119 For MALDI Spotting applications we recommend using the MALDI spotting tip This tip prevents that the droplet from flowing upwards on the outer capillary wall 1 Remove all trays and move the fraction collector arm to the Change Parts position see Maintenance Functions on page 75 Turn the well plate adapter 90 degrees and remove it from the capillary carrier arm Slide the spotting adapter onto the capillary carrier arm and turn it 90 degrees to fixate it The capillary length should be aligned as displayed in Figure 4 Attach the MALDI spotting tip to the capillary The MALDI spotting tip should be aligned to from a flat surface with the capillary tip For correct alignment leave the spotting tip in the plastic strip and push the spotting tip onto the capillary Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Configuration and Operation of the Micro scale Fraction Collector Spotter 2 Configuration of the Micro scale Fraction Collector Spotter Spotting Adapter Triin Fraction Collector Capillary MALDI Spotting Tip Figure 4 Capillary Align
11. 1 5 2 0 ml 96 Agilent plate for AP MALDI 100 Applied Biosystems 2x96 Applied Biosystems 192 Applied Biosystems 400 Perseptive Biosystems e Micromass 80 96 spots Bruker Anchor Chips 384 1536 spots 4 3 for Bruker Anchor Chip 1536 Typically 2 ul depending on the fraction collection container 100 5000 nl depending on the MALDI plate 20 spots min 1spot 3s 100 ul min Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Specifications 6 Table 19 Performance Specifications Agilent 1260 Infinity Micro scale Fraction Collector Spotter G1364D continued Type Specification delay volumes ul 25 um ID fraction collector capillary 0 25 50 um ID fraction collector capillary 1 100 um ID fraction collector capillary 5 cooling Recommended with additional G1330B maximum capacity 2 micro scale fraction collector spotter connected via 2 Position 6 Port micro valve G1162A GLP features Early maintenance feedback EMF electronic records of maintenance and errors interfaces Controller area network CAN optional LAN or external contacts interface RS232C APG remote for remote start stop signals to from other modules Interface to G1330B Thermostat CAN DC out for operation of Agilent approved external devices like valves Only one type of well plate or MALDI plate can be used at a time in one tray Agilent 1260 Infinity Micro scale Fraction Collector
12. 3 Un clip the cable connector which is located between the plastic clips of the capillary guiding assembly JZ of the IA W Kd ji A 96 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Maintenance and Simple Repairs 4 4 Disconnect the lamp assembly from the extension cable at 5 In case the extension cable needs to be replaced as well the cable connector and pull the cable through the guiding pull the extension cable out and push the small connector hole in the carrier assembly of the new one through the hollow shaft of the z arm guiding hole 6 Carefully bend up the two metal sheets of the lamp assembly that hold the LEDs Then slide the assembly onto the spotting adapter that the t shaped alignment plate fits into the slit between the pusher and the adapter Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 97 4 Maintenance and Simple Repairs Connect the lamp assembly to the extension cable Push the cable connector in between the yellow plastic clip as described in step 3 of this procedure Plug the lamp power cable into the connector at the front of the z arm Install the fraction collector spotter capillary as described in Replacing Micro scale Fraction Collector Spotter Capillary Assembly on page 100 To activate the lamp switch open the command line in the ChemStation and type microafclampon to switch the
13. Figure 1 The Agilent 1260 Infinity Micro scale Fraction Collector Spotter In order to ensure reproducible fraction collection and MALDI spotting without cross contamination and an accurate control of the collected volumes the liquid contact control mode as illustrated in Figure 2 on page 38 has been developed During the entire collection process the instrument ensures that the droplet at the capillary tip is in constant contact with the surface of the well the MALDI plate or the liquid being collected At the start of fraction collection the droplet is delivered at the bottom of the well or the MALDI plate While the well is filled the capillary tip moves continuously upwards keeping in constant contact with the liquid surface At the end of fraction collection the capillary tip moves sharply upwards in order to cut the contact between the liquid and the capillary During the collection the flow rate and the geometry of the well are taken into account to calculate the current fill level Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 37 2 Configuration and Operation of the Micro scale Fraction Collector Spotter Introduction Figure 2 Liquid Contact Control 38 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Configuration and Operation of the Micro scale Fraction Collector Spotter 2 Initialization and Reset After power on the instrument delays the initialization for approximately tw
14. installed trays from the tray base Turn OFF the instrument 2 Remove the screw cap from the waste port 3 Exchange the Flap Septum reassemble and tighten the screw cap Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 105 4 Maintenance and Simple Repairs 4 To exchange the waste tubing turn the internal tray upside On completion of this procedure down and unscrew the waste tube fitting from the waste port Assemble the new waste tubing Install the internal tray in the fraction collector as shown in step 4 on page 108 IMPORTANT Make sure that the waste tubing is guided correctly through the waste tubing channel to the waste tubing outlet as shown in step 4 and step 5 on page 108 106 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Maintenance and Simple Repairs 4 Exchanging the Internal Tray Frequency When defective Tools required None Parts required Internal Tray micro scale G1364 63125 Before beginning this procedure 1 Locate the internal tray assembly with the sensor pad and ae waste port in the bottom of the right front corner of the Position the transport unit of the micro instrument collector spotter in the Home Position see Maintenance Functions on page 75 Remove all installed trays from the tray base Turn OFF the instrument Agilent
15. low delay volumes high position accuracy of the capillary tip liquid contact control for reproducible collection of small fractions without carryover The principles of operation for the Agilent 1260 Infinity Micro scale Fraction Collector Spotter will be illustrated in this chapter along with detailed descriptions on how to set up the method and change the configuration This will enable the user to run the instrument with optimal performance The Agilent 1260 Infinity Micro scale Fraction Collector Spotter designed for low flow rates from below 100 ul min down to 100nl min In order to keep dispersion a single low volume PEEK coated fused silica capillary is used to direct the flow to the vessel or MALDI target A permanent waste position is built in to allow automated and continuous collection of the eluent The proper alignment of the capillary tip is crucial for precise collection of the fractions especially if 384 conical well plates or MALDI targets are used A sensor pad for automated control of the capillary tip position was therefore integrated into the micro scale fraction collector spotter A diagram of the fraction collector which indicates the most important components is shown in Figure 1 on page 37 36 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Configuration and Operation of the Micro scale Fraction Collector Spotter 2 A Waste port Sensor pad Capillary Carrier
16. 3 0 The Contact Control adjustment can also be used to optimize the MALDI spotting process and if the flow is splitted before the fraction collector Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Configuration and Operation of the Micro scale Fraction Collector Spotter 2 Auxiliary In the Auxiliary section the Maximum fill volume per location can be specified If as configured is selected the pre configured volume see Instrument gt Pre configured Wellplate Types is used This ensures that the location well vial or tube cannot be overfilled during fraction collection This volume can be further reduced by defining a customized volume By default all MALDI plates are configured with an infinite fill volume The Maximum fill volume can be used to determine a maximum spot size if the peak based trigger mode is used Additional check boxes in this section provide the opportunity to Store the temperature signal and the UIB signal Fraction Preview To determine the appropriate fraction collection parameters the Agilent ChemStation provides a valuable tool that becomes accessible by selecting the button labelled Fraction Preview Tool Figure 7 in the Peak Detectors section Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 53 2 54 Configuration and Operation of the Micro scale Fraction Collector Spotter Setting up a Micro scale Fraction Collector Spotter Method
17. 50 um i d capillary G1364 87305 delay volume app 1 uL for overall flow rates LC flow matrix flow gt 8 ul min the black 100 um ID capillary G1364 87306 delay volume app 4 uL The exchange procedure is described in Replacing Micro scale Fraction Collector Spotter Capillary Assembly on page 100 An Exchange of the fraction collector capillary might also be necessary if the capillary is bent or blocked by particles Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Configuration and Operation of the Micro scale Fraction Collector Spotter 2 Adjusting the Peakwidth of Your Detector When using capillaries in these flow ranges for peak based fraction collection a Peakwidth of 0 1 min Responsetime lt 2s or less should be selected to ensure a fast signal processing Otherwise the compound might already be flushed through the micro scale fraction collector spotter when the peak is detected If a smaller peakwidth is set the flow rate ranges can be increased Capillary Alignment for Micro scale Fraction Collection During the initialization process the vertical and the horizontal position of the capillary tip is determined The micro scale fraction collector spotter facilitates the capillary check by pressing the capillary onto the sensor pad Then the relative capillary length as well as the degree to which the end of the capillary is bent is determined 384 well plates require a higher position accu
18. Fraction Collector Spotter User Manual 31 1 Installing the Micro scale Fraction Collector Spotter Table 7 Configuring Well plate Types Location Description Definition Limits Rows Number of rows on the plate up to 16 Columns Number of columns on the plate up to 24 Volume Volume in pl of a sample vessel A Row distance Distance in mm between the center of two rows B Column distance Distance in mm between the center of two columns C Plate length X size in mm at the bottom ofthe 127 75 0 25 mm plate SBS Standard D Plate width Y size in mm at the bottom of the 85 50 0 25 mm plate SBS Standard E Plate height Size in mm from the bottom to the upto 47mm top of the plate If you use well plates with inlets caps and septa you have to specify the distance from the bottom to the top of the caps F Column offset Distance in mm from the left edge bottom to the center of the first hole A1 G Row offset Distance in mm from the back edge bottom to the center of the first hole A1 H Column shift Offset in mm to Y when the rows are not straight but staggered J Well depth Distance in mm from the top of the up to 45 mm plate to the bottom of the well If you use well plates with inlets caps and septa you have to specify the distance from the top of the septa to the bottom of the inlets 32 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Installing the Micro scale Fraction Colle
19. Internal Tray 107 5 Parts and Materials 109 Supported Trays for the Micro scale Fraction Collector Spotter 110 List of Recommended Plates 112 MALDI Spotting Accessories 113 Supported MALDI Targets 115 Transport Unit Assembly 117 Internal Tray Assembly 118 Micro scale Fraction Collector Spotter Accessory Kit 119 6 Specifications 121 Performance Specifications for the Micro Collector Spotter 122 A Safety Information 125 Safety Information 126 General 126 Operation 127 Safety Symbols 128 Lithium Batteries Information 129 Radio Interference 130 Test and Measurement 130 Sound Emission 131 Manufacturer s Declaration 131 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 7 Contents Solvent Information 132 Solvents 132 Agilent Technologies on Internet 133 8 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Agilent 1260 Infinity Series Micro scale Fraction Collector Spotter User Manual 1 Installing the Micro scale Fraction Collector Spotter Site Requirements 10 Unpacking the Fraction Collector 15 Optimizing the Stack Configuration 18 Installing the Micro scale Fraction Collector Spotter 20 Installing a Thermostatted Fraction Collector 24 Micro scale Fraction Collector Spotter Trays 28 Configure Well plate Types 30 Transporting the Micro scale Collector Spotter 34 ee Agilent Technologies 1 Installing the Micro scale Fraction Collector Spotter Site Require
20. Micro scale Fraction Collector Spotter Configuration dialog 44 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Configuration and Operation of the Micro scale Fraction Collector Spotter 2 Trays In the online ChemStation the configuration of the Trays is recognized automatically In the offline ChemStation the tray configuration can be chosen in a dropdown menu Fraction Delay Volumes The fraction delay volumes specify the volume between the detector cell and fraction collector capillary tip This volume is required to calculate the time delay between the detection of the peak in the detector and the start of the collection in the fraction collector For the micro scale fraction collector spotter G1364D this volume depends on the installed capillaries The fraction delay volume can be calculated by fraction delay volume volume detector outlet capillary volume fraction collector inlet capillary The volumes of the most common Agilent 1260 Infinity detector outlet capillaries and the fraction collector inlet capillaries are summarized in Table 1 on page 45 and Table 2 on page 46 To identify the capillary check the label for the part number internal diameter and capillary length For time based operation the detector cell can be bypassed to minimize the delay volume Table 1 Detector Outlet Capillaries description part number length mm ID um Volume nl DAD MWD outlet capillary for 500 nl flow cell
21. Plug for waste container 1 61364 26105 Reorder gives 5m t Reorder gives 10 clamps for corrugated waste tubing large and 10 clamps for waste tubing small Reorder gives 10 clamps for corrugated waste tubing large and 10 clamps for waste tubing small e The Accessory kit G1330 68705 shown in Table 6 is shipped with the G1330B thermostat module if the thermostat module was ordered Table 6 Optional Thermostat Module Accessory Kit Contents G1330 68705 Description Quantity Part Number Waste tubing 1 2 m 1 5062 2463 Cable thermostat module to micro scale fraction 1 G1330 81600 collector spotter Reorder gives 5m Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 17 1 Installing the Micro scale Fraction Collector Spotter Optimizing the Stack Configuration If your Agilent 1260 Infinity Micro scale Fraction Collector Spotter is part of a system you can ensure optimum performance and minimum delay volume by installing the following configuration Figure 1 on page 19 shows the configuration recommended for the micro scale fraction collector spotter within an Agilent 1260 Infinity Capillary or Nanoflow LC system 18 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Solvent Compartment Micro Degasser Capillary Nano Pump Micro Well Plate Sampler Sampler Thermostat Installing the Micro scale Fraction Collector Spotter 1 Optimizing the Stack Configurati
22. Table4 Delivery Checklist for the G1330B Thermostat Module optional Description Quantity Thermostat Module 1 Power cord local Matching the sockets in your country or region 1 Accessory kit thermostat module see below 1 Accessory Kits Each shipment contents an Accessory kit with the necessary tools to install the system and to have an operating system e The Accessory kit G1364 68715 shown in Table 5 is shipped with the Agilent 1260 Infinity Micro scale Fraction Collector Spotter G1364D Table5 Micro scale Fraction Collector Spotter Accessory Kit Contents G1364 68715 Description Quantity Part Number Hex key 2 0 mm 1 8710 2476 MALDI Spotting Adapter 1 G1364 83205 Flap Septum 1 G1364 27107 Waste Tubing 1 4 mm ID 1 G1364 86711 Waste tubing 1 2 m 1 5062 2463 CAN cable 1m 1 5181 1519 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Installing the Micro scale Fraction Collector Spotter 1 Table5 Micro scale Fraction Collector Spotter Accessory Kit Contents G1364 68715 Description Quantity Part Number Sticking clamp for corrugated waste tubing large 3 5065 9976 Sticking clamp for waste tubing small 3 5065 9976 Micro scale collector spotter Capillary Assembly 25 um ID 1 61364 87304 Micro scale collector spotter Capillary Assembly 100 um ID 1 G1364 87306 ZDV Union stainless steel 1 5022 2184 Strip with 10 MALDI Spotting Tips 1 G1364 81701 Plastic Spacers 20 not orderable
23. Trays Installing the Fraction Collector Trays 1 Press the front door latch holding button located at the front of the right side cover 2 Lift the front door Load the micro scale fraction collector spotter tray with 96 or 384 well plates vial plates Eppendorf plates or MALDI plate holders as required 4 Slide the micro scale fraction collector spotter tray into the micro scale fraction collector spotter so that the rear of the tray is seated firmly against the rear of the tray area Installed trays are automatically detected and identified 5 Press the front of the micro scale fraction collector spotter tray down to secure the tray in the micro scale fraction collector spotter 6 Close the front door If the tray pops out of position the air channel adapter is not correctly inserted Before starting a run the instrument has to be correctly configured in the user interface 28 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Installing the Micro scale Fraction Collector Spotter 1 Numbering of Vial Well plate and Spot Positions With the 4 plates full tray Plate in the left front position P1 Plate in the left back position P2 Plate in the right front position P3 Plate in the right back position P4 Vessel Al A2 B1 B2 With the 2 plates 10 x 2 ml vials std trays Plate in the front position P1 Plate in the back position P2 Vessel Al A2 B1 BQ Vials 1 10 With
24. and Next Location Then the fraction collection is started at the next free plate or at the next free location If no fraction start location has been specified the fraction collection starts at first empty location on the well plate or MALDI target Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 57 2 Configuration and Operation of the Micro scale Fraction Collector Spotter Setting up a Micro scale Fraction Collector Spotter Method Sequence Parameters Instrument 1 x Operator Name Bio Chemist Data File Bar Code Reader Auto Prefix Counter F Use In Sequence Prefix Counter On a bar code mismatch SIG1 0001 Inject anyway Subdirectory PROTEOME Don t inject Path C HPCHEM 1 DATA Part of methods to run gt Shutdown Post Sequence Cmd Macro According to Runtime Checklist SA F Use Sequence Table Information macro SHUTDOWN MAC go v ganume min nRdy Timeout min after loading a new method Fraction Information Fraction Start Locatic Sequence Comment p Cancel Help Figure 10 Start Location in Sequence Parameters dialog box To start a single run e click on the Start button in the graphical user interface of the ChemStation select Run Method from the RunControl menu e press F5 To start the sequence e click on the Start button in the graphical user interface of the ChemStation e s
25. box in the RunControl menu in the Sequence Parameters Figure 10 dialog box or in the Sequence Table in the Sequence menu All fraction start locations are entered in the format micro scale fraction collector spotter plate number row column e g 1 P1 A 01 with micro scale fraction collector spotter 1 2 plate number P1 P4 row is A to H and column 1 12 for 96 well plates or the Agilent AP MALDI plate for different well plates or MALDI plates refer to the description from the plate manufacturer Note the orientation of the MALDI plate The starting position A1 is always in the back left corner of the plate 56 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Confeguredona and oe of the a soale hara Sania 2 Sample Info Instrument 1 p a Micro scale ter Method ollector xi Operator Name Bio Chemist p Data File C Prefix Counter Manual Filename IDEFAULT D Subdirectory PROTEOME Path C HPCHEM 1 DATA r Sample Parameters Fraction Start Sample Name Sample_01 Locatio ication 1 P1 A 01 Sample Amount fo Mana PO O ISTD Amou 0 Dilution Figure 9 Sample Info dialog box run if no entry All settings made in the Sequence Table overrule the settings chosen in the Sequence Parameters screen In addition to the exact position for fraction collection start in the Sequence Table it is also possible to specify Next Plate
26. connections 23 capacities 122 capacity 123 capillary 40 cleaning 41 65 92 MALDI spotting 40 micro fraction collection 40 replacing 99 capillary guiding assembly exchange 103 capillary tip 41 position 41 check out procedure fraction collection 66 MALDI spotting 68 ChemStation configuration 44 cleaning the fraction collector 91 collection order 46 condensation 12 configuration 44 collection order 46 fraction delay volumes 45 needle movement 46 trays 45 well plates 47 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual configuring well plate types 30 Contact Control 47 contact control 47 cooling 123 corrugated waste tubing 21 D damaged packaging 15 data analysis 60 delay volumes 123 delivery checklist 15 door lock sensor 74 deactivate 74 E electrical connections thermostat 27 environment 10 12 F fill levels reset 55 fill volume maximum 53 flap septum exchange 105 flow rate 122 fraction collector capillary 99 fraction delay volumes 45 fraction list 60 fraction preview 53 fraction trigger mode 50 fraction vessel capacities and trays 122 fraction volume 122 fuses 10 G GLP features 123 Index half tray combinations 29 home position 76 initialization 39 installing corrugated waste tubing 21 installing the fraction collector 20 interface cables 20 power cable 20 safety 20 installing the thermostatted
27. fraction collector interface cables 24 power cable 24 preparation 24 safety 20 24 instrument status indicator 73 interfaces 123 internet 133 L liquid contact control 37 47 M maintenance 75 maintenance functions 74 home position 76 park arm position 76 parts exchange position 76 transport unit self alignment 77 maintenance positions 75 MALDI spotting 42 52 MALDI Spotting tip 42 MALDI targets 47 manual reset 39 maximum capacity 123 maximum flow rate 122 135 method 48 setup 48 49 minimum fraction volume 122 missing parts 15 needle assemblies 117 needle movement 46 0 Online 59 operating modes 122 P park arm position 76 park transport unit assembly 34 parts and materials 15 109 needle assemblies 117 plates 112 transport unit assembly 117 parts exchange position 76 peak based 50 peakwidth 41 Performance 122 performance specifications 121 analytical scale 122 physical specifications 13 fraction collector 13 thermostat 13 plates 112 position accuracy calibration 41 79 power consideration 10 power cords 11 power requirements 10 power supply indicator 72 preview 53 recovery 52 recovery on the track 52 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual removing tray compartment divider 26 repairing the fraction collector 89 repairs cleaning the fraction collector 91 main procedures overview 9
28. pre G1315 87338 120 100 945 mounted DAD MWD outlet capillary for 500 nl flow cell G1315 87328 120 50 235 alternative DAD MWD outlet capillary for 500 nl flow cell pre G1315 68708 700 75 3100 mounted old flow cell kit DAD MWD outlet capillary for 80nl flow cell pre G1315 87328 120 50 235 mounted DAD MWD outlet capillary for 80nl flow cell G1315 87318 600 25 300 alternative Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 45 2 Configuration and Operation of the Micro scale Fraction Collector Spotter Table 2 Micro scale Fraction Collector Spotter Inlet Capillaries description part number length mm ID um Volume nl Micro scale fraction collector spotter capillary 25 um G1364 87304 500 25 250 ID alternative Micro scale fraction collector spotter capillary 50m G1364 87305 500 50 1000 ID pre mounted Micro scale fraction collector spotter capillary G1364 87306 500 100 4000 100m ID alternative 46 Due to the tolerance for the capillary ID small adjustments in delay volume might be necessary in order to optimize the performance of your system Collection Order The Collection Order describes capillary movement during fraction collection Four different strategies are possible row by row and column by column either in one direction or in two directions Collection Mode Selection of the Collection Mode is not available for the micro scale fraction collector spotter and therefor
29. procedures 5 Parts and Materials This chapter contains selected illustrations and lists for identification of parts and materials that are required for maintenance or exchanged in simple repair procedures 6 Specifications This chapter contains performance specifications of the micro collector spotter A Safety Information This appendix provides a safety summary Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Contents Contents 1 Installing the Micro scale Fraction Collector Spotter 9 Site Requirements 10 Power Consideration 10 Power Cords 11 Bench Space 12 Environment 12 Unpacking the Fraction Collector 15 Damaged Packaging 15 Delivery Checklist 15 Accessory Kits 16 Optimizing the Stack Configuration 18 Installing the Micro scale Fraction Collector Spotter 20 Installing a Thermostatted Fraction Collector 24 Micro scale Fraction Collector Spotter Trays 28 Installing the Fraction Collector Trays 28 Numbering of Vial Well plate and Spot Positions 29 Configure Well plate Types 30 Transporting the Micro scale Collector Spotter 34 2 Configuration and Operation of the Micro scale Fraction Collector Spotter 35 Introduction 36 Initialization and Reset 39 Configuration of the Micro scale Fraction Collector Spotter 40 Choosing and Installing the Micro scale Fraction Collector Spotter Capillary 40 Configuration
30. revision A 10 02 or higher To activate the lamp switch open the command line in the ChemStation and type microafclampon to switch the lamp ON microafclampoff to switch the lamp OFF Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 39 2 Configuration and Operation of the Micro scale Fraction Collector Spotter Configuration of the Micro scale Fraction Collector Spotter 40 Choosing and Installing the Micro scale Fraction Collector Spotter Capillary The micro scale fraction collector spotter is delivered with three capillaries each having a different inner diameter 25 50 and 100 um The 50 um capillary is installed in the micro scale fraction collector spotter whereas the 25 and the 100 um capillaries are in the accessory kit see Micro scale Fraction Collector Spotter Accessory Kit on page 119 To optimize the micro scale fraction collector spotter for different flow ranges the correct capillary has to be selected and installed For micro scale fraction collection we recommend flow rates below 4 uL min the yellow 25 um i d capillary G1364 87304 delay volume app 0 25 uL flow rates between 4 and 30 uL min the green 50 um i d capillary G1364 87305 delay volume app 1 uL flow rates between 30 and 100 uL min the black 100 um i d capillary G1364 87306 delay volume app 4 uL For MALDI spotting we recommend for overall flow rates LC flow matrix flow lt 8 ul min the green
31. steel probably acts as a catalyst occurs quickly with dried chloroform if the drying process removes the stabilizing alcohol Chromatographic grade ethers which can contain peroxides for example THF dioxane di isopropylether such ethers should be filtered through dry aluminium oxide which adsorbs the peroxides Solutions of organic acids acetic acid formic acid and so on in organic solvents For example a 1 solution of acetic acid in methanol will attack steel Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Safety Information A e Solutions containing strong complexing agents for example EDTA ethylene diamine tetra acetic acid e Mixtures of carbon tetrachloride with 2 propanol or THF Agilent Technologies on Internet For the latest information on products and services visit our worldwide web site on the Internet at http www agilent com Select Products Chemical Analysis It will provide also the latest firmware of the Agilent 1260 Infinity modules for download Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 133 A Safety Information Agilent Technologies on Internet 134 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Index A accessory kit 119 accessory kit contents 17 Agilent on internet 133 air circulation 12 arm length calibration 79 battery safety information 129 bench space 12 C cable
32. 122 atte Agilent Technologies 121 6 Specifications Performance Specifications for the Micro Collector Spotter 122 Table 19 Performance Specifications Agilent 1260 Infinity Micro scale Fraction Collector Spotter G1364D Type Specification trigger modes operating modes fraction vessel capacities and trays MALDI Spotting plates pre configured MALDI Plate Capacity Minimum fraction volume MALDI spot size maximum spotting rate Maximum flow rate Time slices Peak threshold up downslope Timetable combination of time intervals and peak and Agilent 1260 Infinity UV Vis detectors DAD G1315C D G4212B MWD G1365 C D are fully supported Other detectors with appropriate delay volumes can be connected through UIB interface Above location Into location Liquid Contact Control The tip of the fraction collector capillary will initially move down to the bottom of the well Then it will slowly move upwards while the fraction is collected The contact control mode enables the micro scale fraction collector spotter to collect fractions down to 2 ul in well plates or MALDI spots down to 100 nl 4Awell plates full tray MTP with 384 or 96 well plates standard and conical shape or 4 x 27 Eppendorf tubes 0 5 1 5 2 0 ml MALDI Target Plates e 2x well plates std tray MTP 10 x 2 ml vials 1 half tray with 384 or 96 well plates standard and conical shape or 2x 27 eppendorf tubes 0 5
33. 1260 Infinity Micro scale Fraction Collector Spotter User Manual 107 4 Maintenance and Simple Repairs 2 Remove the internal tray by pushing down the plastic holder that holds it in position underneath the metal latch 1 and sliding the tray to the left at the same time 2 3 Remove the corrugated waste tubing from the front of the instrument 1 and slide the internal tray s waste tubing to the rear of the outlet 2 the before sliding the tray out i a eee Kena 4 Install the new tray by sliding it into position underneath the metal leash that holds it The waste tubing from the internal tray should be guided as shown 5 Make sure that the waste tubing is slid all the way through the outlet 1 Its end should be over the edge and below the level of the laboratory desk that the system stands on to avoid any back flow of solvent Re install the corrugated waste tubing 2 On completion of this procedure Re install the tray s in the tray base Start the instrument Close the front cover 108 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Agilent 1260 Infinity Series Micro scale Fraction Collector Spotter User Manual 5 Parts and Materials Supported Trays for the Micro scale Fraction Collector Spotter 110 List of Re
34. 2 simple repairs 90 report 61 reserved locations 48 reset 39 S safety information on lithium batteries 129 sensor pad 36 shipping 34 simple repairs 90 92 site requirements 10 slope only 50 specifications 13 121 spotting adapter 42 spotting rate 51 start location 56 status indicators 72 step commands 87 storage 12 T temperature 12 thermostat electrical connections 27 threshold upper 50 tick marks 61 time based 51 timetable 50 51 transport 34 transport unit assembly 34 117 transport unit self alignment 77 transporting the fraction collector 34 tray combinations 29 tray compartment divider 26 Index trays 45 122 numbering of vessel positions 29 trigger modes 122 troubleshooting and test functions 71 U unpacking the fraction collector 15 upper threshold 50 wW waste port cleaning 92 waste tubing exchange 105 weight 12 well plates 29 47 112 384 conical well plates 79 136 www agilent com In This Book This manual contains user information about the Agilent 1260 Infinity Micro scale Fraction Collector Spotter The manual describes the following e configuration and operation e troubleshooting and test functions e maintenance and simple repairs e parts and materials e specifications e safety information Agilent Technologies 2010 2013 Printed in Germany 10 2013 G1364 90013 Rev B
35. 6 G2255 68700 Only one type of well plates can be used at a time in one tray mixtures in the instrument If you are using flammable solvents remove the plates from the micro collector spotter after turning it OFF You avoid the risk of building explosive gas 112 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Parts and Materials 5 MALDI Spotting Accessories Table 12 MALDI Plate Carriers Plate Type Part Number MALDI Plate Carrier for Agilent Technologies AP MALDI Targets 5022 6543 MALDI Plate Carrier for Bruker AnchorChip 5022 6541 MALDI Plate Carrier for Bruker Prespotted AnchorChip 5022 6546 MALDI Plate Carrier for Applied Biosystems Targets standard format 5022 6542 MALDI Plate Carrier for Applied Biosystems Opti TOF MTP format 5023 0238 MALDI Plate Carrier for Micromass Targets 5022 6544 Table 13 Calibration Plates Plate Type Part Number Calibration Plate for Agilent Technologies AP MALDI Targets 5023 0214 Calibration Plate for Bruker Targets 5023 0208 Calibration Plate for Applied Biosystems Targets 10x10 20x20 5023 0213 Calibration Plate for Applied Biosystems Targets 192 96x2 5023 0209 Calibration Plate for Micromass Targets 5023 0215 Table 14 Online Matrix Kit G1364 68706 Item Part Number External Contact BCD Board G1351 68701 Cable BCD Board to Syringe Pump 5181 1536 Micro Connector T Type 5042 8519 Agilent 1260 Infinity Micro scale Fraction Co
36. G notice denotes a hazard It calls attention to an operating procedure practice or the like that if not correctly per formed or adhered to could result in personal injury or death Do not proceed beyond a WARNING notice until the indicated condi tions are fully understood and met Agilent 1260 Infinity Micro scale Fraction Collector Spotter In This Guide This manual contains user information about the Agilent 1260 Infinity Micro scale Fraction Collector Spotter G1364D It is intended for users that will operate the Agilent 1260 Infinity Micro scale Fraction Collector Spotter either for micro scale fraction collection or for MALDI Spotting For information about installation advanced troubleshooting repair and a complete list of internal parts please refer to the Agilent 1260 Infinity Fraction Collectors Service Manual G1364 90111 1 Installing the Micro scale Fraction Collector Spotter This chapter provides information about the installation of the Agilent 1260 Infinity Micro scale Fraction Collector Spotter 2 Configuration and Operation of the Micro scale Fraction Collector Spotter This chapter describes the configuration method setup and operation of the micro collector spotter 3 Troubleshooting and Test Functions This chapter describes the modules built in troubleshooting test and maintenance functions 4 Maintenance and Simple Repairs This chapter contains instructions on maintenance and simple repair
37. Spotter User Manual 123 6 Specifications Performance Specifications for the Micro Collector Spotter 124 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Agilent 1260 Infinity Series Micro scale Fraction Collector Spotter User Manual A Safety Information Safety Information 126 Lithium Batteries Information 129 Radio Interference 130 Sound Emission 131 Solvent Information 132 Agilent Technologies on Internet 133 ee Agilent Technologies 125 A Safety Information Safety Information The following general safety precautions must be observed during all phases of operation service and repair of this instrument Failure to comply with these precautions or with specific warnings elsewhere in this manual violates safety standards of design manufacture and intended use of the instrument Agilent Technologies assumes no liability for the customer s failure to comply with these requirements General This is a Safety Class I instrument provided with terminal for protective earthing and has been manufactured and tested according to international safety standards WARNING If you are using flammable solvents remove the well plates from the tray when you turn off the sampler You avoid the risk of building explosive gas mixtures in the tray compartment WARNING After a leak in the sampler make sure the leak plane is cleaned and dry 126 Agilent 1260 Infinity Micro scale Fract
38. Troubleshooting and Test Functions 3 Instrument Status Indicator The instrument status indicator indicates the current instrument condition When the status indicator is OFF and power switch light is on the instrument is in a prerun condition and is ready to begin an analysis A green status indicator indicates the instrument is performing an analysis run mode A yellow status indicator indicates a not ready condition The instrument is in a not ready state when it is waiting for a specific condition to be reached or completed for example front door not closed or while a self test procedure is running The next programmed injection will be inhibited until the not ready condition has been cleared An error condition is indicated when the status indicator is red An error condition indicates the instrument has detected an internal problem which affects correct operation of the instrument Usually an error condition requires attention for example leak defective internal components An error condition always interrupts the analysis and prevents the next run after the current run is finished A flashing yellow status indicator indicates that the module is in the resident mode Call your local service provider for assistance upon observing this error condition A flashing red status indicator indicates an during the startup procedure of the module Call your local service provider for assistance upon observing this error condi
39. all flow rates LC flow and matrix flow gt 8 ul min The peek coated fused silica capillary 125 um ID 550mm length is used to deliver the matrix solution from the syringe to the T type connector where it is merged with the flow from the Agilent 1260 Infinity LC System The maximum spot size strongly depends on the type of the MALDI plate and can range between 0 5 and 5 ul The flow rate has to be adjusted to optimize the performance with spotting rate of 20 spots min maximum flow rate maximum spot size x 20 spots min Table5 Examples for Maximum Flow Rates maximum spot size maximum spotting rate maximum flow rate LC flow matrix flow 0 5 ul 20 spots min 10 ul min Iul 20 spots min 20 ul min 2 ul 20 spots min 40 ul min 5 ul 20 spots min 100 L min 64 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Configuration and Operation of the Micro scale Fraction Collector Spotter 2 Contact Control Adjustment If the matrix is added online the flow that is delivered by the Agilent 1260 Infinity Capillary or Nanoflow Pump is different from the flow at the capillary tip in the MALDI spotter To account for this additional flow the Contact Control Adjustment has to be specified as described on page 52 CAUTION If solvents with high concentrations of matrix buffer or salt are used the capillary has to be flushed thoroughly with salt free water after analysis Such a procedure prevents the capilla
40. aste tubing The plug G1364 26105 is included in the micro scale fraction collector spotter accessory kit es UVDetector Pump Control Cable T lype Connector Syringe Pump ee ee a es Matrix Capillary SS Be nj eh er me MALDI Spotter Figure 14 Online Matrix Delivery Syringe Pump Control The Online Matrix Kit contains an External Contact Board BCD G1351 68701 to control the syringe pump In addition a cable to connect the KDS 200 syringe pump is provided 5181 1536 To connect other Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Configuration and Operation of the Micro scale Fraction Collector Spotter 2 syringe pumps a general purpose control cable 18594 60520 can be ordered and configured The BCD board can be installed in any Agilent 1260 Infinity LC module For installation details refer to the corresponding service manual The pump control can easily be setup in the ChemStation as displayed in Figure 15 To open the dialog box select the module where the external contact board is installed and click on the icon in the graphical user interface Then select Contacts The default setting should be set as open i e the syringe pump is switched off In the example that is shown in Figure 15 the syringe pump is switched off by default Using the Timetable the matrix flow can be switched as required during the run We recommend switching the
41. ation and type microafclampon to switch the lamp ON microafclampoff to switch the lamp OFF During the position accuracy calibration the door lock sensor is disabled The fraction collector arm will move while the front door is open In addition the side panel can be removed during this procedure To avoid personal injury keep fingers away from the needle area during position accuracy calibration All users must be informed about this potential risk 82 To perform a position accuracy calibration for well plates Step Note 1 Switch to the Diagnosis View of the Agilent ChemStation Software 2 Select Tests from the Diagnosis menu 3 Inthe Test Selection dialog box select Micro AFC G1364D from the drop down list Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Troubleshooting and Test Functions 3 To perform a position accuracy calibration for well plates continued Step Note 4 Select position accuracy calibration well plates and click on Start 5 Start the position accuracy calibration and Click on Explain to get detailed instructions follow the instructions on the screen and information 6 Align the capillary tip to the center of each See Figure 19 on page 83 The height corner well with the corresponding positioning calibration is not required for well plates and buttons therefore greyed out 7 Repeat Step 6 for all configured plates Needle Adjustment Plate 1 x
42. ay 107 ee Agilent Technologies 89 4 Maintenance and Simple Repairs Introduction into Repairing the Micro scale Fraction Collector Spotter Simple Repairs The micro collector spotter is designed for easy repair The most frequent repairs and maintenance tasks such as changing the capillary the flap septum or the waste tubing can be done from the front of the instrument with the instrument in place in the system stack These repairs are described in Simple Repairs Procedures on page 99 WARNING When opening capillary or tube fittings solvents may leak out Please observe appropriate safety procedures for example goggles safety gloves and protective clothing as described in the material handling and safety data sheet supplied by the solvent vendor especially when toxic or hazardous solvents are used WARNING Regularly inspect the capillary and exchange it if they is worn out or shows visible signs of damage WARNING To prevent personal injury the power cable must be removed from the instrument before removing the micro collector spotter cover Do not connect the power cable to the micro collector spotter while the cover is removed 90 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Maintenance and Simple Repairs 4 Cleaning the Micro Collector Spotter The micro collector spotter covers should be kept clean Cleaning should be done with a soft cloth slightly dampened with wa
43. cale fraction collector spotter is OFF 9 Connect the power cable to the power connector at the rear of the micro scale fraction collector spotter 10 Connect the CAN cable to the other Agilent 1260 Infinity LC modules 11 If an Agilent ChemStation is the controller it must be connected to the LAN interface should be installed to the detector 12 Connect the APG remote cable optional for non Agilent 1260 Infinity instruments 13 Install the tray that has been delivered for your micro scale fraction collector spotter Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 21 1 22 Installing the Micro scale Fraction Collector Spotter The micro scale fraction collector spotter G1364D is shipped with 4 x well plate full tray All other trays have to be ordered separately 14 Turn ON power by pushing the button at the lower left hand side of the micro scale fraction collector spotter 15 The exhaust fan will turn ON and remove potential solvent vapor from the inside of the instrument After 2 minutes close the front door Then the micro scale fraction collector spotter will start the hardware initialization process At the end of this process the status LED should be green Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Installing the Micro scale Fraction Collector Spotter 1 CAN cable to previous module
44. calibration plate is not available use the MALDI Target and align the capillary to the center of each corner position 7 Align the height z position of the capillary tip The capillary tip should be aligned just above the MALDI Target leaving enough space for sheet of paper to fit between the capillary tip and the target 8 Repeat Step 6 7 for all configured targets Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 85 3 86 Troubleshooting and Test Functions Position Accuracy Calibration for MALDI Targets Needle Adjustment Plate 1 xy position alignment buttons height z position alignment buttons Only available for MALDI targets Corrective Values relative to the previous alignment Figure 20 Position Accuracy Calibration Interface for MALDI Targets Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Troubleshooting and Test Functions 3 Single Step Commands Some movements of the fraction collection or spotting sequence can be done under manual control This is useful during troubleshooting where close observation of each of the fraction collection steps is required to confirm a specific failure mode or verify successful completion of a repair Each step command actually consists of a series of individual commands which move the micro collector spotter components to predefined positions enabling the specific step to be done In the ChemStation the st
45. ck the More button Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 51 2 52 Configuration and Operation of the Micro scale Fraction Collector Spotter Recovery If the Recovery on the Track mode is checked the solvent that is delivered between the fractions spots is collected on the same plate for recovery The capillary carrier will move to the next well following collection order e g row by row In the Data Analysis view as well as in the Report a fraction list indicates the wells that contain fractions and the wells that contain recovery With this information the user is able to recover compounds that weren t collected Contact Control Adjustment This parameter is only required for MALDI Spotting If you are adding your matrix online as described in Online Matrix Delivery on page 62 the flow rate at the capillary tip of the micro scale fraction collector spotter is higher than the flow rate that is measured at the flow sensor of your Agilent 1260 Infinity Capillary or Nanoflow Pump Consequently the spotting tip retraction speed has to be accommodated to the higher flowrate The factor can be calculated by Contact Control Adjustment pump flow matrix flow pump flow Some examples are presented in the Table 4 below Table 4 Examples for Contact Control Adjustment Pump Flow ol Matrix Flow nl Contact Control Adjustment 500 0 1 0 default 500 500 2 0 1000 500 1 5 500 1000
46. commended Plates 112 MALDI Spotting Accessories 113 Supported MALDI Targets 115 Transport Unit Assembly 117 Internal Tray Assembly 118 Micro scale Fraction Collector Spotter Accessory Kit 119 s f Agilent Technologies 109 5 Parts and Materials Supported Trays for the Micro scale Fraction Collector Spotter Table 10 Trays for the Micro scale Fraction Collector Spotter Item Description Part Number 1 Full tray for 4 well plates G1364 84521 2 Full tray for 4 MALDI plate carriers adjustable G1364 84511 3 Std tray for 2 well plates 10 2ml vials G1367 60011 4 Adapter air channel installed underneath plug channel if G1329 43200 the fraction collector is used with the thermostat 5 Plug channel G1364 47200 Only one type of well plate can be used at a time 110 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Parts and Materials Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 5 111 5 Parts and Materials List of Recommended Plates Table 11 Recommended Plates Item Description Select in ChemStation Volume ml Package Part Number 1 96 polypropylene well plate 96Agilent 0 5 10 5042 1386 2 96 polypropylene well plate 96Agilent 0 5 120 5042 1385 3 384 polypropylene well plate 384Agilent 0 1 30 5042 1388 4 96 polypropylene conical well plate 96EppendorfC 0 18 25 5042 8502 5 54 x 2ml vial plate 54VialPlate 1 5
47. ctor Spotter 1 Table 7 Configuring Well plate Types continued Location Description Definition Limits Well X size Size of the well in x direction Plate min 3 7 mm length If you use well plates with min 3 0 mm with inlets caps and septa you have to position accuracy specify the x size of the septa alignment micro scale Well Y size Size of the well in y direction Plate down to 3 7 mm width If you use well plates with min 3 0 mm with inlets caps and septa you have to position accuracy specify the y size of the septa alignment micro scale Bottom size For round wells the relative of the 1 0 cylindrical well top and bottom of the well 0 0 conical well Square Click in the field to specify whether Yes rectangular the well is rectangular or round No round oval Is well plate Click in this field to specify if this is Yes well plate or a well plate or not Relevant for continuous flow operation MALDI Target No Vial Tray or Eppendorf tray The distances need to be measured with high precision It is recommended to use a caliper If possible contact the vendor of non predefined well plates to obtain these dimensions Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 33 1 Installing the Micro scale Fraction Collector Spotter Transporting the Micro scale Collector Spotter 34 When moving the Agilent 1260 Infinity Micro scale Fraction Collector Spotter inside the labora
48. e greyed out in this configuration The micro scale fraction collector spotter always operates in the continuous flow mode Needle Movement Into location In the into location mode the capillary tip moves into the well to the specified Depth in mm Above location In the above location mode the capillary tip stays at the specified Distance in mm above the well during fraction collection Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Configuration and Operation of the Micro scale Fraction Collector Spotter 2 Contact Control In this mode the capillary tip moves down until is reaches the specified distance between capillary tip and well bottom This ensures that the forming droplet is in contact with the well bottom During the continuing filling process the capillary tip moves upwards while staying in contact with the surface of the collected liquid For low flow rates lt 1 ul min a small Distance lt 0 2 mm should be chosen This value should be increased to 0 3 0 7 mm in particular for MALDI spotting at higher flow rates The capillary movement in the Contact Control mode depends on the flow rate that is delivered by the Agilent 1260 Infinity pump If your LC system contains more than one pump the flow rate of the first pump in your system determines the needle movement In this case verify in your system configuration Instrument gt Configure 1260 access that this pump is the first one in the Config
49. e position accuracy depends on the temperature We recommend to perform the position accuracy alignment at the same temperature as the operation of the instrument If a fraction collector thermostat is present it should be switched on 30 minutes before the calibration to ensure constant temperature conditions in the module 84 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Step Troubleshooting and Test Functions 3 To perform a position accuracy calibration for MALDI targets Note Switch to the Diagnosis View of the Agilent ChemStation Software Select Tests from the Diagnosis menu In the Test Selection dialog box select Micro AFC G1364D from the drop down list Select position accuracy calibration MALDI and click on Start Start the position accuracy calibration and follow the instructions on the screen Align the xy position of the capillary tip to the center of each corner position of the MALDI Target or to each pinhole on the MALDI calibration plates with the corresponding positioning buttons Click on Explain to get detailed instructions and information See Figure 20 on page 86 for Agilent Technologies AP MALDI targets use calibration plate 5023 0214 for Micromass MALDI targets use calibration plate 5023 0215 for Bruker MALDI Targets use calibration plate 5023 0208 for Applied Biosystems MALDI targets use Calibration plates 5023 0209 or 5023 0213 e Ifa
50. elect Run Sequence from the RunControl menu e press F6 58 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Configuration and Operation of the Micro scale Fraction Collector Spotter 2 Setting up a Micro scale Fraction Collector Spotter Method Online Tick Marks To display tick marks for fraction start and stop events in you Online Plot click on the Change button in your Online Plot window Then check Show fraction collection ticks in the Edit Signal Plot window Figure 11 Instrument 1 Fle unono irstnumerk Method Sequence Pufy View Abot Help ig Methods 4 crecnoutas m Sequence Lit Signal Pion GAB A ignabes4a 16 Retarencen2 100 mau Same sone JE CHECKOU TOON e Ka J me JN Rosy mmn Figure 11 Online Tick Marks Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 59 2 Configuration and Operation of the Micro scale Fraction Collector Spotter Viewing your Results Viewing your Results Data Analysis In order to display the tick marks for the collected fractions or spots on the screen click on Signal options from the Graphics menu Then choose Separated in the Layout drop down menu To review your chromatograms file information and a fraction list select the Data Analysis view from the drop down menu and press the Fraction Task button as displayed in Figure 12 Purification Task Button Chromatogram with Fraction Tick Marks instrum
51. ent 1 a Date Time tho b 1 3 2006 2 31 28 DAD A 5105254 3 RemO0 100 CIEL NTACHECKOUTOORRO D PIAN a 73 E7 Wi a DADI ae 214 i eS DADI 38 Dani 0 0063 min 0 198 one 2141 datapoints Fraction Table Figure 12 Data Analysis View 60 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Configuration and Operation of the Micro scale Fraction Collector Spotter 2 Viewing your Results Report In order to create reports with a fraction table and tick marks the Specify Report dialog box the item Add Fraction Table and Tick Marks has to be checked Specify Report Instrument 1 M File Type wTXT M WMF r DF F csy F xLS HTM Figure 13 Fraction collection report setup Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 61 2 Configuration and Operation of the Micro scale Fraction Collector Spotter Online Matrix Delivery 62 The Agilent 1260 Infinity Micro scale Fraction Collector Spotter can be configured for online matrix delivery For this application the Online Matrix Kit G1364 68706 is required see Table 14 on page 113 This kit contains all accessories required to control a syringe pump and to connect the syringe to the LC flow path An overview is displayed in Figure 14 For low flow rates and for MALDI spotting with online matrix delivery we recommend using the plug to close the waste container instead of the w
52. ep commands can be selected from the Automatic Fraction Collector Step box see Figure 21 in the Diagnosis display In the Agilent Instant Pilot G4208A the step commands can be accessed from the pull down menu in the micro collector spotter Test Automatic Fraction Collector Steps Needle Up Move to Location Vial Needle into Vessel Figure 21 Fraction Collector Step Commands Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 87 3 88 Troubleshooting and Test Functions Table8 Step Commands Step Action Comments Needle Up Lifts the capillary arm to the upper position Move to Location Moves the capillary arm to the set collection spotting location Move to Location Lowers the capillary tip into the specified vessel Move to Waste Moves the needle into the waste port Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Agilent 1260 Infinity Series Micro scale Fraction Collector Spotter User Manual 4 Maintenance and Simple Repairs Introduction into Repairing the Micro scale Fraction Collector Spotter 90 Simple Repairs 90 Cleaning the Micro Collector Spotter 91 Cleaning the Waste Port 92 Replacing the Micro scale Fraction Collector Spotter Lamp Assembly 95 Replacing Micro scale Fraction Collector Spotter Capillary Assembly 100 Exchanging the Capillary Guiding Assembly 103 Exchanging the Flap Septum and the Waste Tubing 105 Exchanging the Internal Tr
53. etable If the LC system is used for analytical chromatography without fraction collection or MALDI spotting the micro scale fraction collector spotter can be switched Off The spot size for MALDI spotting is specified by the flow rate and the collection time If peak based trigger mode is selected for MALDI spotting the collection time and therefore the spot size is flexible In this case we recommend specifying a maximum fill volume per location as described in Auxiliary on page 53 50 Peak Detectors The Peak Detectors section in this screen comprises a list of all Peak Detectors connected to the system As working modes for each peak detector Threshold only Threshold Slope and Slope only are possible In the Threshold only mode the settings for Up Slope Down Slope and Upper Threshold in the subsequent columns are ignored Fraction collection is triggered whenever the detector signal exceeds the specified threshold value When the signal drops below the threshold value fraction collection is stopped In the Slope only mode fraction collection is triggered on the slopes of the detector signals Adequate values for Up Slope and Down Slope can be specified in the corresponding fields In the Threshold Slope mode fraction collection is triggered on the corresponding values for threshold and slope For more detailed information concerning working with threshold and slopes please refer to Agilent s Application Note Sophisticated pea
54. even though the instrument has been disconnected from its source of supply Dangerous voltages capable of causing serious personal injury are present in this instrument Use extreme caution when handling testing and adjusting Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 127 A Safety Information Safety Symbols Table 20 shows safety symbols used on the instrument and in the manuals Table 20 Safety Symbols Symbol Description The apparatus is marked with this symbol when the user should refer to the instruction manual in order to prevent risk of harm to the operator and to protect the apparatus against damage Indicates dangerous voltages Indicates a protected conductor terminal Eye damage may result from directly viewing the light produced by the Xenon flash lamp used in this product Always turn the xenon flash lamp off before removing it Q A warning alerts you to situations that could cause physical injury or damage to the equipment Do not proceed beyond a warning until you have fully understood and met the indicated conditions CAUTION A caution alerts you to situations that could cause a possible loss of data Do not proceed beyond a caution until you have fully understood and met the indicated conditions 128 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Safety Information A Lithium Batteries Information Danger of explosion
55. fractions will be collected based on the peak detection parameters given in the Peak Detector table Additionally a Maximum Peak Duration in minutes has to be specified This parameter can be used to stop the fraction collection in cases where the baseline drifts and the signal does not drop below the specified threshold value The limits are from 0 1 min to 9999 0 min This parameter is mandatory if you use Peak Controlled fraction collection but is disabled for Time Based fraction collection Timetable When the Time based mode is chosen two different options are available e A specific number of fractions or time slices can be specified The of Fractions refer to the number of fractions that are collected in the corresponding time frame in the table This parameter is applicable only for time based fraction collection The maximum spotting rate is 20 spots min e Alternatively time slices can be set up that define the collection time for each fraction This parameter is only available in the time based fraction collection mode Based on the maximum spotting rate of 20 spots per minute the minimum time slice for one spot is 3s e Contact Adj is a parameter which is only required for MALDI Spotting For details please read Contact Control Adjustment on page 52 To edit the Timetable the functions Insert Append Cut Copy and Paste are available To access the additional sections in the Setup Fraction Collector dialog box cli
56. gle injection under the conditions as described in Table 6 You should observe a chromatogram with four peaks and four fractions should have been collected in the well plate Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 67 2 68 Configuration and Operation of the Micro scale Fraction Collector Spotter Check out Procedure for MALDI Spotting To check out the MALDI spotting functionality a run with blank injection with time based spotting is performed to verify that Agilent 1260 Infinity Micro scale Fraction Collector Spotter consistently collects onto the MALDI target Prerequisites V The Agilent 1260 Infinity Capillary LC or Nanoflow System has been installed and prepared as described in the Agilent 1260 Infinity Capillary LC System Manual G1376 90013 or the Agilent 1260 Infinity Nanoflow LC System Quick Reference G2226 90013 V The Agilent 1260 Infinity Micro scale Fraction Collector Spotter has been installed and configured for MALDI spotting The delay volume has been specified The MALDI adapter and the MALDI spotting tip have been installed V The Position Accuracy Calibration for MALDI Targets on page 84 has been executed before running the checkout V A MALDI plate carrier with supported MALDI target is installed as plate 1 in the 4 well plate tray Table 7 Method Parameters for MALDI Spotting Check out Agilent 1260 Infinity modules micro degasser capillary pump 20pl flow senso
57. ide of the fraction collector It can easily be removed by lifting the clip and pulling the whole tray to the left side 3 Now the whole waste vial can be removed as well 93 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 4 Maintenance and Simple Repairs To clean the wash port and the waste tubing For a thorough cleaning of all components they can sonicated for 15 minutes in a ultrasonic bath filled with water Another way to get rid of plugging is to flush the waste capillary from the exit with a water filled syringe Attach the syringe at the end of the waste tubing using a with a 1 6 mm OD capillary and flush the waste tubing and the waste port thoroughly In order to avoid flooding of the fraction collector a tissue should be placed besides the internal tray close to the spill On completion of this procedure Assemble the cleaned waste tubing Install the internal tray in the fraction collector as shown in step 4 on page 108 IMPORTANT Make sure that the waste tubing is guided correctly through the waste tubing channel to the waste tubing outlet as shown in step 4 and step 5 on page 108 94 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Maintenance and Simple Repairs 4 Replacing the Micro scale Fraction Collector Spotter Lamp Assembly Frequency When lamp is defective or not required in the micro scale f
58. if battery is incorrectly replaced Replace only with the same or equivalent type recommended by the equipment manufacturer Lithium batteries may not be disposed off into the domestic waste Transportation of discharged Lithium batteries through carriers regulated by IATA ICAO ADR RID IMDG is not allowed Discharged Lithium batteries shall be disposed off locally according to national waste disposal regulations for batteries Lithiumbatteri Eksplosionsfare ved fejlagtic handtering Udskiftning ma kun ske med batteri af samme fabrikat og type Lever det brugte batteri tilbage til leverandoren Lithiumbatteri Eksplosionsfare Ved udskiftning benyttes kun batteri som anbefalt av apparatfabrikanten Brukt batteri returneres appararleverandoren Bij dit apparaat zijn batterijen geleverd Wanneer deze leeg zijn moet u ze niet weggooien maar inleveren als KCA Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 129 A Safety Information Radio Interference Never use cables other than the ones supplied by Agilent Technologies to ensure proper functionality and compliance with safety or EMC regulations Test and Measurement If test and measurement equipment is operated with equipment unscreened cables and or used for measurements on open set ups the user has to assure that under operating conditions the radio interference limits are still met within the premises 130 Agilent 1260 Infinity Micro scale F
59. illary tip is at the correct position remove the wellplate adapter tighten the clamp nut and remount the adapter O pe clamp nut 0 102 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Maintenance and Simple Repairs 4 Exchanging the Capillary Guiding Assembly Frequency When Capillary guiding assembly is bent or damaged Tools required None Parts required Capillary guiding assembly G1364 87303 WARNING To avoid personal injury keep fingers away from the needle area during micro collector spotter operation CAUTION Regularly inspect the Fraction Collector Capillary and exchange it if it is worn out or shows visible signs of damage Before beginning this procedure 1 Remove the wellplate adapter and the clamp nut Then pull the capillary from the clip at the bottom of the carrier assembly as well as the capillary guiding assembly Move the transport unit of the micro collector spotter in the Home Position see Maintenance Functions on page 75 Remove all installed trays from the tray base Position the transport unit of the micro collector spotter in the Exchange Parts Position see Maintenance Functions on page 75 Turn OFF the instrument IMPORTANT Do not adjust the capillary while the clamp nut is tightly mounted on the capillary guide
60. ion Accuracy Calibration for 384 Well Plates on page 82 have to be executed before running the check out V All wells are empty and the fill levels which are stored on the module have been reset To verify this remove and insert the 4 well plate tray Then click on Yes in displayed dialog box Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Configuration and Operation of the Micro scale Fraction Collector Spotter 2 Table 6 Method Parameters for Micro scale Fraction Collection Check out Agilent 1260 Infinity modules Column Sample Flow Stoptime Solvent A Solvent B Wavelength DAD MWD Peakwidth Responsetime Injection volume Column temperature Fraction trigger mode Max peak duration DAD MWD working mode Upslope Downslope Threshold Upper threshold micro degasser capillary pump 20pl flow sensor micro autosampler micro well plate sampler column compartment optional diode array detector multiple wavelength detector with 500nl or 80 nl flow cell micro scale fraction collector spotter ZORBAX SB C18 5 um 150 x 0 5mm 5064 8256 Check out sample 01080 68704 diluted 1 10 in Acetonitrile 15 pl min 10 minutes 30 HPLC grade water 70 HPLC grade acetonitrile Signal A 254 4nm Reference 360 80nm gt 0 1min 2s 200 nl 25 0 C or ambient Peak based 1 min Threshold slope 5 mAU s 5 mAU s 5 mAU 3000 mAU Expected Result Run a sin
61. ion Collector Spotter User Manual Safety Information A Operation Before applying power comply with the installation section Additionally the following must be observed Do not remove instrument covers when operating Before the instrument is switched on all protective earth terminals extension cords auto transformers and devices connected to it must be connected to a protective earth via a ground socket Any interruption of the protective earth grounding will cause a potential shock hazard that could result in serious personal injury Whenever it is likely that the protection has been impaired the instrument must be made inoperative and be secured against any intended operation Make sure that only fuses with the required rated current and of the specified type normal blow time delay and so on are used for replacement The use of repaired fuses and the short circuiting of fuseholders must be avoided WARNING Any adjustment maintenance and repair of the opened instrument under voltage is forbidden WARNING Disconnect the instrument from the line and unplug the power cord before maintenance Do not operate the instrument in the presence of flammable gases or fumes Operation of any electrical instrument in such an environment constitutes a definite safety hazard Do not install substitute parts or make any unauthorized modification to the instrument Capacitors inside the instrument may still be charged
62. ions move these assemblies into the appropriate maintenance position All trays have to be removed before starting the Maintenance dialog In the ChemStation the micro scale fraction collector spotter maintenance positions can be selected from the Maintenance menu in the Diagnosis view see Figure 17 In the Agilent Instant Pilot G4208A the functions can be selected in the Test screens of the micro scale fraction collector spotter Fraction collector Maintenance Positions Change Parts Move Arm Home Park Arm for Transport Ss Figure 17 Fraction Collector Maintenance Positions dialog box Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 75 3 76 Troubleshooting and Test Functions Change Parts If you click Start the transport unit will move upwards the capillary carrier assembly will move to the front center and then turn off the rotary motor to allow free rotation of the arm This position enables easy access e to change the capillary guiding assembly e to change the micro scale fraction collector spotter capillary e to change the capillary carrier assembly e to change the micro scale fraction collector spotter lamp assembly e to replace the well plate adapter with the spotting adapter or vice versa e to align the capillary length and install the spotting tip After the maintenance or repair task has been finished click End to reset the micro scale f
63. k based fraction collection working with up and down slope publication number 5988 7895EN The option to specify an Upper Threshold becomes important if the absorbance exceeds the linear range of the UV Vis detector At high absorbance values the light intensity on the detector is extremely low and consequently detector noise will superimpose the detector signal In this case the detector noise might lead to wrong trigger commands for the micro scale fraction collector spotter As soon as the detector signal exceeds the Upper Threshold settings for slopes will be ignored until the signal drops again below the Upper Threshold Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Configuration and Operation of the Micro scale Fraction Collector Spotter 2 When using more than one peak detector fraction collection can be triggered either when all selected peak detectors detect a peak or when at least one selected peak detector detects a peak based on the settings in the detector table The Timetable can be used to program changes in the Fraction Trigger Mode during the analysis by entering a time in the Time field and appropriate values in the fields of the timetable The trigger modes can be specified as Off Peak based and Time based Time ranges can be defined for the different trigger modes In the column headed Time If the Off mode is selected no fractions are collected Whenever the Peak based mode is specified
64. lamp ON microafclampoff to switch the lamp OFF 98 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Maintenance and Simple Repairs 4 Simple Repairs Procedures The procedures described in this section can be done with the micro collector spotter in place in the stack Table9 Simple Repair Procedures Procedure Typical Frequency Notes Replacing the fraction collector When worn out blocked or damaged See Replacing Micro scale Fraction capillary When using a capillary with different diameter Collector Spotter Capillary Assembly on page 100 Exchanging the capillary guiding When bent or damaged See Exchanging the Capillary Guiding assembly Assembly on page 103 Exchanging the internal tray When defective See Exchanging the Internal Tray on page 107 Exchanging the flap septum and When defective or contaminated See Exchanging the Flap Septum and the waste tubing the Waste Tubing on page 105 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 99 4 Maintenance and Simple Repairs Replacing Micro scale Fraction Collector Spotter Capillary Assembly Frequency When contaminated blocked worn out or visibly damaged When using a new capillary with different diameter Tools required None Parts Required Capillary Assembly 25 um ID G1364 87304 Capillary Assembly 50 um ID G1364 87305 or Capillary Assembly 100 um ID G1364 87306 WARNING To avoid per
65. le fraction collector spotter all fill levels can be reset by removing and re installing the 4 well plate tray not just the well plate or MALDI plate carrier After re installation of the tray a pop up window as displayed in Figure 8 will be displayed Click on Yes to reset all fill levels from the previous run gt Tray 1 was replaced by a tray of the same type 4 Do you want to reset the fill volume es Figure 8 Reset fill volumes The window Figure 8 to reset current fill levels is not displayed if the well plate or the MALDI target is removed without removing the complete tray Consequently contaminated fractions unwanted overfill events or split fractions might be observed during the following LC run Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 55 2 Configuration and Operation of the Micro scale Fraction Collector Spotter The run time for a system with a fraction collector must be extended by the delay time to ensure the complete collection of all compounds The run time is calculated as run time end of last peak delay time with delay time delay volume flow rate This calculation is only required if the flow rate or the delay volume capillary is changed For further info read Fraction Delay Volumes on page 45 Assignment of Start Location The start location for fraction collection or MALDI spotting can either be assigned in the Sample Info Figure 9 dialog
66. le fraction collector spotter are in use Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 25 Installing the Micro scale Fraction Collector Spotter Figure 5 Removing the Tray Compartment Divider 12 Install the tray you have ordered for your micro scale fraction collector spotter 13 Ensure the power switch on the front of the micro scale fraction collector spotter is OFF and the power cables are disconnected 14 Connect the cable between the micro scale fraction collector spotter and the thermostat see Figure 6 on page 27 Do not disconnect or reconnect the micro scale fraction collector spotter to thermostat cable when the power cords are connected to either of the two modules This will damage the electronics of the modules 26 15 Connect the power cables to the power connectors 16 Connect the CAN cable to other Agilent 1260 Infinity LC modules 17 If an Agilent ChemStation is the controller it must be connected to the LAN interface 18 Connect the APG remote cable optional for non Agilent 1260 Infinity instruments 19 Turn ON power by pushing the button at the lower left hand side of the micro scale fraction collector spotter 20 The exhaust fan will turn ON and remove potential solvent vapor from the inside of the instrument After 2 minutes close the front door Then the Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Installing the Micro scale F
67. le fraction collector spotter dimensions and weight see Table 2 on page 13 allow the instrument to be placed on almost any laboratory bench The instrument requires an additional 25 cm 10 inches of space on either side for the circulation of air and approximately 8 cm 8 1 inches at the rear for electrical connections Ensure the micro scale fraction collector spotter is installed in a horizontal position If a complete Agilent 1260 Infinity LC System is to be installed on the bench make sure that the bench is designed to carry the weight of all the modules For a complete system including the thermostatted micro scale fraction collector spotter it is recommended to position the modules in two stacks see Recommend Stack Configuration for micro scale collector spotter in the capillary or nanoflow system on page 19 Make sure that in this configuration there is 25 cm 10 inches space on either side of the thermostatted micro scale fraction collector spotter for the circulation of air Environment CAUTION Your micro scale fraction collector spotter will work within specifications at ambient temperatures and relative humidity as described in Table 1 and Table 2 on page 13 Do not store ship or use your micro scale fraction collector spotter under conditions where temperature fluctuations may cause condensation within the micro scale fraction collector spotter Condensation will damage the system electronics If your micro scale f
68. llector Spotter User Manual 113 5 Parts and Materials Table 14 Online Matrix Kit G1364 68706 continued Item Part Number Peek coated fused silica capillary 550mm length ID 125t1m G1375 87318 Adapter 10 32 to 1 4 28 5042 8517 Syringe 1ml 5181 1541 Union to Luer Fitting 5042 8518 Needle LL 22 51 3 2 pk 5183 4614 114 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Supported MALDI Targets Table 15 Supported Targets for MALDI Spotting Parts and Materials 5 Note MALDI target plates for Applied Biosystems Bruker and Micromass are not supplied by Agilent Technologies Dimensions for other targets can be configured in the ChemStation Software Agilent Technologies 96 Agilent for AP MALDI G1972 60025 Applied Biosystems 80 Applied Biosystems 96 Applied Biosystems 100 Applied Biosystems 96x2a Applied Biosystems 96x2b Applied Biosystems 192 Applied Biosystems 400 Perseptive Biosystems 144 Applied Biosystems 192 Opti TOF target 384 Opti TOF target MPT format 96 Opti TOF target MPT format Bruker 384 AnchorChip Sample 384 AnchorChip Lock 1536 AnchorChip 384 Prespotted AnchorChip Micromass Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 115 5 Parts and Materials Table 15 Supported Targets for MALDI Spotting continued Note MALDI target plates for Applied Biosystems Bruker and Micromass are not supplied b
69. ment for MALDI Spotting Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 43 2 Configuration and Operation of the Micro scale Fraction Collector Spotter Configuration of the ChemStation To setup or change the configuration parameters of your fraction collector select More Fraction Collector gt Configuration from the Instrument menu or right click on the fraction collector icon in the graphical user interface In the Fraction Collector Configuration dialog box Figure 5 the configuration of the Trays the Fraction Delay Volumes the Collection Order the Needle Movement and the well plate MALDI target type can be specified Fraction Collector Configuration Instrument 1 E xj m Trays Tray A 4 WellPlates M Tube Yolume ml 0 00 Tube Height mm fo Fraction Delay Volumes Collection Order Shortest path O Detector Volume nl Row byrow 3 iby couat c Column by c Shortest path column by row r Collection Mode Needle Movement G pi s C into location Depth 5 0 mm Discrete fractions c P C above location Distance 20 mm Continuous flow contact control Distance 0 2 mm m Well Plates Plate Type 96E ppendorfC z r Tice Reserved Locations Installed Plates Row s IV Plate 1 front left IV Plate 3 front right Column s Plate 2 back left 7 Plate 4 back right anme Location s Cancel Help More gt gt Figure 5
70. ments 10 A suitable site environment is important to ensure optimum performance of the micro scale fraction collector spotter Power Consideration The micro scale fraction collector spotter power supply has wide ranging capability see Table 1 on page 13 Consequently there is no voltage selector in the rear of the micro scale fraction collector spotter There are also no externally accessible fuses as automatic electronic fuses are integrated in the power supply The thermostatted micro scale fraction collector spotter comprises two modules the micro scale fraction collector spotter G1364D and the thermostat G1330B Both modules have a separate power supply and a power plug for the line connections The two modules are connected by a Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Installing the Micro scale Fraction Collector Spotter 1 control cable and both are turned on by the micro scale fraction collector spotter module The thermostat power supply has two externally accessible fuses To disconnect the micro scale fraction collector spotter from line power unplug the power cord The power supply still uses some power even if the power switch on the front panel is turned off To disconnect the thermostatted micro scale fraction collector spotter from line power unplug the power cord from the micro scale fraction collector spotter and the thermostat The power supplies still use some power e
71. o minutes to vent the fraction compartment Then the door lock is activated and the micro scale fraction collector spotter remains in the not ready condition yellow waiting for the manual reset Before you reset the micro scale fraction collector spotter the door has to be closed To reset the instrument click on the micro scale fraction collector spotter icon in the graphical user interface of the ChemStation and start the Reset If the main pump is switched ON a dialog is displayed which reminds the user to switch OFF the pump Then the reset procedure is started without the risk of contaminating well plates or fractions During the initialization process the vertical and the horizontal position of the capillary tip is determined The micro scale fraction collector spotter performs the capillary check by pressing the capillary tip onto the sensor pad After this capillary check a message is displayed if any further capillary alignment is required For more details about capillary alignment refer to Choosing and Installing the Micro scale Fraction Collector Spotter Capillary on page 40 Switching the Micro scale Fraction Collector Spotter Lamp The micro scale fraction collector spotter includes a lamp to illuminate the fraction compartment The lamp is specifically required for the position accuracy calibration as described on page 82 and page 84 The operation of the lamp requires firmware revision A 05 09 or higher and ChemStation
72. o scale Fraction Collector Spotter User Manual Parts and Materials 5 Micro scale Fraction Collector Spotter Accessory Kit Table 18 Micro scale Fraction Collector Spotter Accessory Kit Contents G1364 68715 Description Quantity Part Number Hex key 2 0 mm 1 8710 2476 MALDI Spotting Adapter 1 G1364 83205 Flap Septum 1 G1364 27107 Waste Tubing 1 4 mm ID 1 G1364 86711 Waste tubing 1 2 m 1 5062 2463 CAN cable 1 m 1 5181 1519 Sticking clamp for corrugated waste tubing large 3 5065 9976 Sticking clamp for waste tubing small 3 5065 9976 Fraction Collector Capillary Assembly 25 um ID 1 G1364 87304 Fraction Collector Capillary Assembly 100 um ID 1 G1364 87306 ZDV Union stainless steel 1 5022 2184 Strip with 10 MALDI Spotting Tips 1 G1364 81701 Plastic Spacers 20 not orderable Plug for waste container 1 G1364 26105 Reorder gives 5 m t Reorder gives 10 clamps for corrugated waste tubing large and 10 clamps for waste tubing small Reorder gives 10 clamps for corrugated waste tubing large and 10 clamps for waste tubing small Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 119 5 Parts and Materials Micro scale Fraction Collector Spotter Accessory Kit 120 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Agilent 1260 Infinity Series Micro scale Fraction Collector Spotter User Manual 6 Specifications Performance Specifications for the Micro Collector Spotter
73. of the ChemStation 44 Setting up a Micro scale Fraction Collector Spotter Method 49 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 5 Contents Fraction Preview 53 Starting Your Run with Fraction Collection or MALDI Spotting 55 Assignment of Start Location 56 Online Tick Marks 59 Viewing your Results 60 Data Analysis 60 Report 61 Online Matrix Delivery 62 Check out Procedures 66 Check out Procedure for Micro scale Fraction Collection 66 Check out Procedure for MALDI Spotting 68 Troubleshooting and Test Functions 71 Status Indicators 72 Power Supply Indicator 72 Instrument Status Indicator 73 Deactivating the Door Lock Sensor 74 Maintenance Functions 75 Transport Unit Self Alignment 77 Arm Length Calibration 79 Position Accuracy Calibration for 384 Well Plates 82 Position Accuracy Calibration for MALDI Targets 84 Single Step Commands 87 Maintenance and Simple Repairs 89 Introduction into Repairing the Micro scale Fraction Collector Spotter 90 Simple Repairs 90 Cleaning the Micro Collector Spotter 91 Cleaning the Waste Port 92 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Contents Replacing the Micro scale Fraction Collector Spotter Lamp Assembly 95 Simple Repairs Procedures 99 Replacing Micro scale Fraction Collector Spotter Capillary Assembly 100 Exchanging the Capillary Guiding Assembly 103 Exchanging the Flap Septum and the Waste Tubing 105 Exchanging the
74. on z a Column Compartment INET f J UV Vis detector i NED C Micro i collector spotter as 4 n Fraction Collector Thermostat rg Figure 1 Recommend Stack Configuration for micro scale collector spotter in the cap illary or nanoflow system The micro scale fraction collector spotter should never be positioned on top of a module that generates heat which can lead to an unwanted evaporation of fractions in the micro scale fraction collector spotter e g Agilent 1260 Infinity Thermostatted Column Compartment G1316A or Agilent 1260 Infinity Diode Array Detectors Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 19 1 Installing the Micro scale Fraction Collector Spotter Installing the Micro scale Fraction Collector Spotter Preparation Locate bench space Provide power connections Unpack the micro scale fraction collector spotter Parts required Fraction Collector Power cord Chemstation or Agilent Instant Pilot G4208A WARNING When opening capillary or tube fittings solvents may leak out Please observe appropriate safety procedures for example goggles safety gloves and protective clothing as described in the material handling and safety data sheet supplied by the solvent vendor especially when toxic or hazardous solvents are used WARNING To avoid personal injury keep fingers away from the needle area during micro scale fraction collector spotter
75. operation Do not attempt to insert or remove a vial ora plate when the needle is positioned 1 Install the LAN interface board in the micro scale fraction collector spotter Gf required Remove the adhesive tape which covers the side and front doors Open the front door and remove the left side door Remove the transport protection foam oF WwW N Install the corrugated waste tube in the plastic port at the front bottom center of the instrument Slide the waste tubing coming from the internal tray if present through the plastic port and the corrugated waste tube see Figure 2 on page 21 Route the corrugated waste tubing into a waste container 20 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Installing the Micro scale Fraction Collector Spotter 1 Figure 2 Installing the Corrugated Waste Tubing in the Plastic Port 6 Re install the left side door take care of the magnet at the back Ensure the side door is correctly installed its presence is sensed by a hall sensor a missing side door will result in a NOT READY state of the instrument 7 Place the micro scale fraction collector spotter in the stack or on the bench in all horizontal position 8 Ensure the power switch at the front of the micro s
76. or spotter has been mechanically and electrically checked If the shipping container or cushioning material is damaged notify the carrier and keep the shipping material for the carrier s inspection If there are signs of damage to the micro scale fraction collector spotter please do not attempt to install the micro scale fraction collector spotter Delivery Checklist Ensure all parts and materials have been delivered with the micro scale fraction collector spotter For this compare the shipment content with the checklist included in each instrument box Please report missing or damaged parts to your local Agilent Technologies sales and service office The Agilent 1260 Infinity Micro scale Fraction Collector Spotter is available as e G1364D micro scale fraction collector spotter designed for flow rates below 100 ul min and for use with well plates and Eppendorf tubes G1364D Thermostatted option of the micro scale fraction collector spotter This option can be setup by additionally ordering and installing a G1330B fraction collector thermostat Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 15 1 16 Installing the Micro scale Fraction Collector Spotter Table 3 Delivery Checklist for the G1364D Micro scale Fraction Collector Spotter Description Quantity Micro scale collector spotter unit 1 Power cord local Matching the sockets in your country or region 1 Accessory kit see below 1
77. ots should be consistently aligned on the MALDI target Due to evaporation the spots collected earlier during the run will be smaller than the later spots Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 69 2 Configuration and Operation of the Micro scale Fraction Collector Spotter Check out Procedures 70 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Agilent 1260 Infinity Series Micro scale Fraction Collector Spotter User Manual 3 Troubleshooting and Test Functions Status Indicators 72 Deactivating the Door Lock Sensor 74 Maintenance Functions 75 Transport Unit Self Alignment 77 Arm Length Calibration 79 Position Accuracy Calibration for 384 Well Plates 82 Position Accuracy Calibration for MALDI Targets 84 Single Step Commands 87 s j Agilent Technologies 71 3 Troubleshooting and Test Functions Status Indicators Two status indicators are located at the front panel of the micro collector spotter The lower left indicates the power supply status the upper right indicates the micro collector spotter status Status indicator green yellow red Line power switch with green light Figure 16 Location of Status Indicators Power Supply Indicator The power supply indicator is integrated into the main power switch When the indicator is illuminated green the power is ON 72 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual
78. r or nanoflow pump micro autosampler micro well plate sampler column compartment optional diode array detector multiple wavelength detector with 500nl or 80 nl flow cell optional micro scale fraction collector spotter configured for MALDI Spotting Column ZORBAX SB C18 5 pm 150 mm x 0 5 mm 5064 8256 for the capillary pump ZORBAX SB C18 3 5 um 150 mm x 100 um 5065 9910 or similar column for the nanoflow pump Sample blank injection no sample required Flow 5 pL min capillary pump or 1 pL min nanoflow pump Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Configuration and Operation of the Micro scale Fraction Collector Spotter 2 Table 7 Method Parameters for MALDI Spotting Check out continued Stoptime 3 minutes capillary pump 6 minutes nanoflow pump Solvent A 30 HPLC grade water Solvent B 70 HPLC grade acetonitrile Injection volume blank injection Column temperature 25 0 C or ambient Fraction trigger mode Use Timetable 1 With the capillary pump Time 0 5 minutes Trigger mode Time based of fractions 20 Time 2 5 minutes Trigger mode Off 2 With the nanoflow pump Time 0 5 minutes Trigger mode Time based of fractions 10 Time 5 5 minutes Trigger mode Off Expected Result Run a single injection under the conditions as described in Table 7 You should observe 20 spots with the capillary LC system or 10 spots with a nanoflow LC system The sp
79. r take care of the magnet at the back Ensure the side door is correctly installed its presence is sensed by a hall sensor a missing side door will result in a NOT READY state of the instrument 9 Place the micro scale fraction collector spotter on top of the thermostat Make sure that the micro scale fraction collector spotter is correctly engaged in the thermostat locks 10 Remove the plastic cover from the tray base place the air channel adapter 1 into the micro scale fraction collector spotter tray base Make sure the adapter is fully pressed down This assures that the cold airstream from the thermostat is correctly guided to the tray area of the micro scale fraction collector spotter Place the plug channel 2 on top of the air channel adapter Both devices must be installed correctly to assure proper operation of the instrument Figure 4 Installation of Thermostat and Fraction Collector 11 The Agilent 1260 Infinity Micro scale Fraction Collector Spotter is delivered with a pre installed tray compartment divider This divider should only be used with the thermostatted micro scale fraction collector spotter if a half or std tray is installed into the left and center positions of the instrument If any tray is installed to the right side of the instrument remove the tray compartment divider The tray compartment divider optimizes the cooling performance of the instrument if only the left and center position of the micro sca
80. raction collector spotter Tools required None Parts Required Micro scale fraction collector spotter lamp assembly G1364 60014 WARNING To avoid personal injury keep fingers away from the capillary arm during micro collector spotter operation WARNING Explicitly follow the described installation procedures to maximize the lifetime of the fraction collector capillary and to avoid potential spills or fraction losses Regularly inspect the capillary and exchange it if it is worn out or shows visible signs of damage Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 95 4 Maintenance and Simple Repairs Before beginning this procedure Position the transport unit of the micro collector spotter in the Home Position see Maintenance Functions on page 75 Remove all installed trays Position the transport unit of the micro collector spotter in the Change Parts Position see Maintenance Functions on page 75 Turn OFF the instrument Unscrew the micro collector spotter capillary from the flow cell of the detector Remove the fraction collector spotter capillary as described in Replacing Micro scale Fraction Collector Spotter Capillary Assembly on page 100 1 Unplug the lamp power cable from the connector at the front of the z arm 2 To disassemble the lamp carefully bend up the two metal sheets that hold the LEDs Then push up the lamp assembly
81. raction Collector Spotter 1 micro scale fraction collector spotter will start the hardware initialization process At the end of this process the status LED should be green 21 The micro scale fraction collector spotter is turned ON when the line power switch is pressed and the green indicator lamp is illuminated The detector is turned OFF when the line power switch is protruding and the green light is OFF WARNING To disconnect the micro scale fraction collector spotter from the line unplug the power cord The power supply still uses some power even if the power switch at the front panel is turned OFF WARNING To avoid personal injury keep fingers away from the needle area during micro scale fraction collector spotter operation Do not attempt to insert or remove a vial or a plate when the needle is positioned aain Fraction Collector Thermostat cable HEBBE 0006006 CAN bus AC Power Figure 6 Connections at the Rear of the Thermostatted Fraction Collector Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 27 1 Installing the Micro scale Fraction Collector Spotter Micro scale Fraction Collector Spotter
82. raction Collector Spotter User Manual Safety Information A Sound Emission Manufacturer s Declaration This statement is provided to comply with the requirements of the German Sound Emission Directive of 18 January 1991 This product has a sound pressure emission at the operator position lt 70 dB e Sound Pressure Lp lt 70 dB A e At Operator Position e Normal Operation e According to ISO 7779 1988 EN 27779 1991 Type Test Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 131 A Safety Information Solvent Information Observe the following recommendations on the use of solvents This instrument should only be used with solvents that have an ignition temperature higher than 200 C 132 Solvents Brown glass ware can avoid growth of algae Always filter solvents small particles can permanently block the capillaries Avoid the use of the following steel corrosive solvents Solutions of alkali halides and their respective acids for example lithium iodide potassium chloride and so on High concentrations of inorganic acids like nitric acid sulfuric acid especially at higher temperatures replace if your chromatography method allows by phosphoric acid or phosphate buffer which are less corrosive against stainless steel Halogenated solvents or mixtures which form radicals and or acids for example 2CHCl3 Og gt 2COCl 2HCl This reaction in which stainless
83. raction collector spotter and move the arm to the waste port Home Position This maintenance function moves the arm up and to the right rear for better access and exchange of the trays Park Arm This maintenance position moves the arm to the park position at the upper rear left side of the tray for transporting or shipping the micro collector spotter Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Troubleshooting and Test Functions 3 Transport Unit Self Alignment The transport unit alignment is required to compensate for larger deviations in positioning the needle carrier assembly This might be necassary after disassembling the system or when you exchange the transport unit the capillary carrier assembly or the MTP main board This function is in the diagnose screen of the ChemStation or the Agilent Instant Pilot G4208A WARNING The sample transport self alignment requires the 4 well plate tray G1364 84511 or G1364 84521 All well plates MUST be removed If the Transport Unit Self Alignment is started with well plates on the tray the alignment procedure is aborted WITHOUT error message When is a Transport Unit Self Alignment Necessary The sample transport self alignment is required after disassembling the module or when you exchange e The transport unit e The capillary carrier assembly e The MTP main board How to perform a Transport Unit Self Alignment Steps Comments
84. raction collector spotter was shipped in cold weather leave it in its box and allow it to warm up slowly to room temperature to avoid condensation 12 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Installing the Micro scale Fraction Collector Spotter 1 Table 1 Physical Specifications Micro scale Fraction Collector Spotter G1364D Type Specification Comments Weight 13 5 kg 29 8 Ibs Dimensions 200 x 345 x 440 mm height x width x depth 8 x 13 5 x 17 inches Line voltage 100 240 VAC 10 Wide ranging capability Line frequency 50 or 60 Hz 5 Power consumption apparent power 200 VA Maximum Power consumption active power 180 W Maximum Ambient operating temperature 4 45 C 41 113 F Ambient non operating temperature 40 70 C 4 158 F Humidity lt 95 at 25 40 C 77 104 F Non condensing Operating Altitude Up to 2000 m 6500 ft Non operating altitude Up to 4600 m 14950 ft For storing the micro scale fraction collector spotter Safety standards IEC CSA UL Installation Category II Pollution Degree 2 For indoor use only Table2 Physical Specifications Thermostat G1330B Type Specification Comments Weight 18 5 kg 40 7 Ibs Dimensions 140 x 345 x 435 mm height x width x depth 5 5 x 13 5 x 17 inches Line voltage 100 120 or 220 240 VAC 10 Wide ranging capability Line frequency 50 or 60 Hz t 5 Power cons
85. racy than 96 well plates Consequently a slightly bent capillary might be tolerated for 96 well plates whereas 384 well plates require the installation of a new capillary or a position accuracy calibration as described in Position Accuracy Calibration for 384 Well Plates on page 82 After the capillary check procedure a message is displayed if any further capillary alignment is required CAUTION If solvents with high concentrations of matrix buffer or salt are used the capillary has to be flushed thoroughly with salt free water after analysis Such a procedure prevents the capillary and the waste port from clogging The capillary length has to be adjusted between the following limits see Figure 3 For 384 conical well plates PCR plates where high position accuracy is required the capillary tip should be approximately set to the groove at the center of the well plate adapter When 96 well plates or deep 384 well plates are used the capillary tip must be set to the lower edge of the well plate adapter opening in order to reach the bottom of the wells Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 41 2 42 Configuration and Operation of the Micro scale Fraction Collector Spotter high position accuracy maximum capillary length e g 384 well plates e g deep well plates
86. ration of the Micro scale Fraction Collector Spotter equal quarters the order of the four plates is the same as displayed in the Installed Plates section Each tray position that contains a well plate has to be checked In the Reserved Locations you have the possibility to specify locations that will not be used for Fraction Collection see Table 3 Table 3 Syntax for the Definition of Reserved Locations Location Syntax Description Rows A Row A can t be used A B Rows A and B can t be used A D A B C and D can t be used A D F Rows A B C D and F can t be used Columns 1 Column 1 can t be used 1 2 Columns 1 and 2 can t be used 1 4 Columns 1 2 3 and 4 can t be used 1 4 12 Columns 1 2 3 4 and 12 can t be used Single locations G12 H12 Locations G12 and H12 can t be used Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Configuration and Operation of the Micro scale Fraction Collector Spotter Setting up a Micro scale Fraction Collector Spotter Method To setup the method parameters of the micro scale fraction collector spotter select Setup Fraction Collector from the Instrument menu or right click on the fraction collector icon in the graphical user interface This will open the Setup Fraction Collector dialog box as displayed in Figure 6 In the Setup Fraction Collector dialog box general method settings are specified Set up Fraction Collector Instrument 1 E i
87. rol Technology Licenses The hardware and or software described in this document are furnished under a license and may be used or copied only in accor dance with the terms of such license Restricted Rights Legend If software is for use in the performance of a U S Government prime contract or subcon tract Software is delivered and licensed as Commercial computer software as defined in DFAR 252 227 7014 June 1995 or asa commercial item as defined in FAR 2 101 a or as Restricted computer soft ware as defined in FAR 52 227 19 June 1987 or any equivalent agency regulation or contract clause Use duplication or disclo sure of Software is subject to Agilent Tech nologies standard commercial license terms and non DOD Departments and Agencies of the U S Government will receive no greater than Restricted Rights as defined in FAR 52 227 19 c 1 2 June 1987 U S Government users will receive no greater than Limited Rights as defined in FAR 52 227 14 June 1987 or DFAR 252 227 7015 b 2 November 1995 as applicable in any technical data Safety Notices CAUTION A CAUTION notice denotes a haz ard It calls attention to an operat ing procedure practice or the like that if not correctly performed or adhered to could result in damage to the product or loss of important data Do not proceed beyond a CAUTION notice until the indicated conditions are fully understood and met A WARNIN
88. ry and the waste port from clogging Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 65 2 Configuration and Operation of the Micro scale Fraction Collector Spotter Check out Procedures 66 The purpose of the check out injection is the verification of the correct installation and configuration of the Agilent 1260 Infinity Micro scale Fraction Collector Spotter system In addition the checkout procedure can used for troubleshooting if the Agilent 1260 Infinity Micro scale Fraction Collector Spotter doesn t operate as expected The checkout procedure depends on the application and is different for micro fraction collection into well plates and for MALDI spotting Check out Procedure for Micro scale Fraction Collection The check out procedure is based on the procedure for the Agilent 1260 Infinity Capillary LC System After the check out sample 01080 68704 has been injected four peaks have to be detected and four fractions collected Prerequisites V The Agilent 1260 Infinity Capillary LC System has been installed and prepared as described in the Agilent 1260 Infinity Capillary LC System Manual G1376 90013 V The Agilent 1260 Infinity Micro scale Fraction Collector Spotter has been installed and configured The delay volume has been specified V A well plate is installed as plate 1 in the 4 well plate tray If 384 well plates are used the Arm Length Calibration on page 79 and the Posit
89. sonal injury keep fingers away from the capillary arm during micro collector spotter operation WARNING Thoroughly follow the described installation procedures to maximize the lifetime of the fraction collector capillary and to avoid potential spills or fraction losses Regularly inspect the capillary and exchange it if it is worn out or shows visible signs of damage Before beginning this procedure Position the transport unit of the micro collector spotter in the Home Position see Maintenance Functions on page 75 Remove all installed trays from the tray base Position the transport unit of the micro collector spotter in the Change Parts Position see Maintenance Functions on page 75 Turn OFF the instrument Unscrew the micro collector spotter capillary from the flow cell of the detector 1 Remove the wellplate adapter from the Capillary Carrier Assembly 100 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Maintenance and Simple Repairs 4 2 Remove the clamp nut and remove the capillary from the clip at the bottom of the carrier assembly Do not adjust the capillary while the clamp nut is tightly mounted on the capillary guide Eg E ULE 3 Pull the capillary out off the capillary guiding pipe and out off the guiding hole in z arm of the transport assembly Do no
90. syringe pump on at least 10 minutes before the start of the LC run in order to prime the matrix capillary and ensure that the matrix is added to the LC flow immediately after the run is started Contact 1 is open syringe pump is switched off Contact 1 is closed syringe pump is switched on xi r Contact 1 Contact 2 Contact 3 Contact 4 open open open open closed closed C closed closed m Timetable Time min C1 c2 c3 C4 Insert 1 0 00 Append _ 2 20 00 open pe Cut Copy Paste OK Cancel Help Figure 15 Syringe Pump Control through External Contacts Capillaries and Fittings The Online Matrix Kit contains Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 63 2 Configuration and Operation of the Micro scale Fraction Collector Spotter e peek coated fused silica capillary 125 um ID 550mm length G1375 87318 e T type connector 5042 8519 e Connector syringe to capillary 5042 8517 To setup the flow connections the outlet capillary from the UV detector the fraction collector capillary and the capillary from the matrix pump have to be connected to the T type Connector as displayed in Figure 14 on page 62 We recommend using e the 50 um ID fraction collector capillary G1364 87305 for overall flow rates LC flow and matrix flow lt 8 ul min e the 100 um ID fraction collector capillary G1364 87306 for over
91. t tighten the clamp nut without a capillary installed M hole 2 guiding pipe 4 Press the metal sheets of fixation spring together and push the capillary assembly out of the holder at the z arm of the transport assembly 5 Unplug the capillary assembly from the bolt carrier located on the top center of the front cover by moving the snapper as indicated by the arrows Slide the Capillary out to the bottom Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 101 4 Maintenance and Simple Repairs 6 Push the new capillary through the bolt carrier as shown in step 5 of this procedure Then attach the fixation spring at the z arm of the transport assembly and push the capillary through the guiding hole 7 Push the capillary through the guiding fitting assembly and attach the wellplate adapter Then adjust the position of the capillary tip For highest accuracy that is required for 384 well plates the tip must be close to the groove in the middle of the wellplate adapter 8 If you are using a deep well plate you should push the capillary through the guiding assembly that the tip is close to the bottom of the wellplate adapter frame The tip must stay within the oval frame 9 If the cap
92. tainer Make sure the condensation drain tube runs down into a waste container without any upwards bends or curves Free and unrestricted flow of the condensation into a waste container must be guaranteed Make sure that the condensation drain tube is always above the liquid level in the container If the tube is located in liquid the condensed water cannot flow out of the tube and the outlet is blocked Any further condensation will then remain in the instrument This may damage the instruments electronics CAUTION The micro scale fraction collector spotter thermostat requires 25 cm 10 inch space on each for sufficient air circulation 24 3 Install the LAN interface board in the micro scale fraction collector spotter Gf required Remove the adhesive tape which covers the side and front doors Open the front door and remove the left side door Remove the transport protection foam xn Oo Oo A Install the corrugated waste tube in the plastic port at the front bottom center of the micro scale fraction collector spotter and route down into a waste container Slide the waste tubing coming from the internal tray Gf present through the plastic port and the corrugated waste tube see Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Installing the Micro scale Fraction Collector Spotter 1 Figure 2 on page 21 Route the corrugated waste tubing into a waste container 8 Re install the left side doo
93. ter or a solution of water and a mild detergent Do not use an excessively damp cloth from which liquid could drip into the micro collector spotter WARNING Do not let liquid drip into the micro collector spotter It could cause a shock hazard or damage to the micro collector spotter Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 91 4 Maintenance and Simple Repairs Cleaning the Waste Port Frequency The frequency depends strongly on the application Tools required None Parts Required None To avoid personal injury keep fingers away from the capillary arm during micro collector spotter operation CAUTION If solvents with high concentrations of matrix buffer or salt are used the capillary has to be flushed thoroughly with salt free water after analysis Such a procedure prevents the capillary and the waste port from clogging 92 For low flow rates and for MALDI spotting with online matrix delivery we recommend to use the plug to close the waste container instead the waste tubing The plug G1364 26105 is included in the micro scale fraction collector spotter accessory kit Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 4 Maintenance and Simple Repairs 2 Turn the tray upside down unscrew the tubing and remove the washer 4 After unscrewing the cap the waste port can be further disassembled 1 The internal tray is located on the right s
94. the 100 vials std tray Vial 1 100 With the half trays Left hand 40 position tray 1 40 Center 40 position tray 101 140 Right hand 40 position tray 201 240 or Left hand 15 position tray 1 15 Center 15 position tray 101 115 Right hand 15 position tray 201 215 Never use Contact Mode or Into Location Mode with 2 mL vials 15 mL vials or deep wellplates The wellplate adapter will dip into the liquid and or the spotter capillary may be too short to touch the liquid Do not use closing mats The micro scale fraction collector spotter has no needle therefore the capillary will be bend or clogged after several injections Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 29 1 Installing the Micro scale Fraction Collector Spotter Configure Well plate Types 30 If the plate you are using is not found on the List of Recommended Plates on page 112 you may configure a custom plate Measure the exact dimensions of the plate as marked below and enter the values in the plate configuration table of the ChemStation or the Agilent Instant Pilot G4208A lt a ae Ja PS ic Figure 7 Well plate Dimensions straight Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Installing the Micro scale Fraction Collector Spotter 1 Configure Well plate Types Figure 8 Well plate Dimensions staggered Agilent 1260 Infinity Micro scale
95. tion Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 73 3 Troubleshooting and Test Functions Deactivating the Door Lock Sensor The Agilent 1260 Infinity Micro scale Fraction Collector Spotter G1364D allows the user to operate the instrument with an open front door In order to use this option the door lock sensor has to be disabled Type the following command into the ChemStation Command Line doorunlock The door lock sensor should only be disabled for special applications It should be activated again if this operation mode is no longer necessary To activate the door lock sensor again type doorlock If the door lock sensor is disabled the instrument will stay in the READY status while the door is open However the fraction collector will always startup in the default mode door lock on after it has been power cycled If the door lock sensor is disabled the fraction collector arm will move while the front door is open To avoid personal injury keep fingers away from moving parts during fraction collector operation All users must be informed about this potential risk 74 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Troubleshooting and Test Functions 3 Maintenance Functions Some maintenance procedures require the fraction collector spotter arm and capillary carrier to move to specific positions to enable easy access to these components The maintenance funct
96. tion Open the command line in the ChemStation and type microafclampon to switch the lamp ON microafclampoff to switch the lamp OFF WARNING During the arm length calibration the door lock sensor is disabled The fraction collector arm will move while the front door is open In addition the side panel can be removed during this procedure To avoid personal injury keep fingers away from the needle area during arm length calibration All users must be informed about this potential risk To perform an arm length calibration for 384 well plates Step Note 1 Before starting the arm length calibration procedure put two 384 well plates into the front positions 1 and 3 of the well plate tray Select the type of these plates in the fraction collector configuration dialog 2 Switch to the Diagnosis View of the Agilent ChemStation Software 3 Select Tests from the Diagnosis menu 4 Inthe Test Selection dialog box select Micro AFC G1364D from the drop down list Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 79 3 80 Troubleshooting and Test Functions To perform an arm length calibration for 384 well plates continued Step Note 10 11 Select arm length calibration 384 well plates and click on Start Start the arm length calibration and follow the instructions on the screen Click the Plate 1 button in the Arm Length Calibration dialog box Figure 18 on page 81
97. tory no special precautions are needed However if the micro scale fraction collector spotter needs to be shipped to another location via carrier ensure V The transport assembly is in the park position Use the ChemStation or the Agilent Instant Pilot G4208A for this command To move the arm to the park position 1 Switch to the Diagnosis view of the ChemStation and select Fraction Collector gt Maintenance Positions from the Maintenance menu 2 In the upcoming dialog box click Park Arm V The vial tray and the sample transport mechanism is secured with the transport protection foam Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Agilent 1260 Infinity Series Micro scale Fraction Collector Spotter User Manual 2 Configuration and Operation of the Micro scale Fraction Collector Spotter Introduction 36 Initialization and Reset 39 Configuration of the Micro scale Fraction Collector Spotter 40 Setting up a Micro scale Fraction Collector Spotter Method 49 Viewing your Results 60 Check out Procedures 66 Agg Agilent Technologies 35 2 Configuration and Operation of the Micro scale Fraction Collector Spotter Introduction The Agilent 1260 Infinity Micro scale Fraction Collector Spotter G1364D is an instrument for micro scale fraction collection or MALDI Spotting with the Agilent 1260 Infinity Capillary or Nanoflow LC Systems With these systems the collection of very low volume fractions requires
98. umption active power 210 W Maximum Power consumption apparent power 260 VA Maximum Ambient operating temperature 4 40 C 41 131 F Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 13 1 Installing the Micro scale Fraction Collector Spotter Table2 Physical Specifications Thermostat G1330B continued Type Specification Comments Ambient non operating temperature Humidity Operating Altitude Non operating altitude Safety standards IEC CSA UL EN 40 70 C 4 158 F lt 95 at 25 40 C 77 104 F Non condensing Up to 2000 m 6500 ft Up to 4600 m 14950 ft For storing the micro scale fraction collector spotter Installation Category II Pollution Degree 2 For indoor use only 14 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Installing the Micro scale Fraction Collector Spotter 1 Unpacking the Fraction Collector CAUTION If you need to ship the micro scale fraction collector spotter at a later date always use the shipping protection foam parts see Transporting the Micro scale Collector Spotter on page 34 Damaged Packaging CAUTION Upon receipt of your micro scale fraction collector spotter inspect the shipping containers for any signs of damage If the containers or cushioning material are damaged keep them until the contents have been checked for completeness and the micro scale fraction collect
99. ured Modules list In addition this pump should also be selected as main pump Instrument gt Change main pump Well Plates In the Well Plates section the type of well plates or MALDI targets used in a well plate tray can be configured The well plate type used can be chosen from the Plate Type dropdown menu More detailed information about all preconfigured well plates and MALDI targets can be found in the Instrument menu A list of recommended Agilent well plates can be found in Table 11 on page 112 and a list of supported MALDI targets is available in Table 15 on page 115 To configure other well plates or MALDI targets choose Configure Well Plates from the Instrument menu Note the orientation of the well plate or the MALDI plate The starting position A1 is always at the back left corner of the plate If the well plate tray is loaded with multiple well plates or vial plates all plates have to be of the same type e g 4 x Agilent conical 96 well plate The checkbox Treat as 4x 96 is only available for 384 well plates and allows splitting the 384 well plate virtually into 4 separate 96 well plates This only changes the collection order but the numbering of the wells remains as indicated on the plate The filling order of each quarter is as specified in the Collection Order section When the 384 well plate is split into four Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 47 2 48 Configuration and Ope
100. ven if the power switch on the front panel is turned off Please make sure that it is always possible to access the power plug Shock hazard or damage of your instrumentation can result if the devices are connected to a line voltage higher than specified Power Cords Your micro scale fraction collector spotter is delivered with a power cord which matches the wall socket of your particular country or region The plug on the power cord which connects to the rear of the instrument is identical for all types of power cord Never operate your instrumentation from a power outlet that has no ground connection Never use a power cord other than the power cord designed for your region Never use cables other than the ones supplied by Agilent Technologies to ensure proper functionality and compliance with safety or EMC regulations Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 11 1 Installing the Micro scale Fraction Collector Spotter Bench Space The micro scale fraction collector spotter dimensions and weight see Table 1 on page 13 allow the instrument to be placed on almost any laboratory bench The instrument requires an additional 2 5 cm 1 0 inch of space on either side and approximately 8 cm 3 1 inches at the rear for the circulation of air and room for electrical connections Ensure the micro scale fraction collector spotter is installed in a horizontal position The thermostatted micro sca
101. y Agilent Technologies Dimensions for other targets can be configured in the ChemStation Software 96 Micromass Sample 96 Micromass Lock 84 Micromass Sample 84 Micromass Lock 84 Micromass Calib 116 Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual Parts and Materials 5 Transport Unit Assembly Table 16 Transport Unit Assembly Micro Scale Item Description Part Number 1 Transport unit assembly micro scale G1364 60020 includes items 2 4 2 Capillary carrier assembly micro scale 61364 60023 3 Capillary Guiding Assembly 61364 87303 4 Wellplate adapter 61364 23203 5a Fraction Collector Capillary Assembly 25 um ID G1364 87304 5b Fraction Collector Capillary Assembly 50 um ID 61364 87305 5c Fraction Collector Capillary Assembly 100 um ID G1364 87306 Figure 23 Transport Unit Assembly Micro Scale Agilent 1260 Infinity Micro scale Fraction Collector Spotter User Manual 117 5 Parts and Materials Internal Tray Assembly Table 17 Internal Tray Assembly Micro Scale Item Description Part Number 1 Internal tray assembly micro scale includes items 2 6 G1364 63125 2 Waste tubing with fitting G1364 86711 3 Washer 3050 2204 4 Screw cap 9301 1379 5 Flap septum G1364 27107 6 Waste Container G1364 22301 l eB O oq m ous 4 Figure 24 Internal Tray Assembly Micro Scale 118 Agilent 1260 Infinity Micr

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