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and Preparative-scale Fraction Collectors

1. 3 Unplug the inlet waste tubing 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 tubings out to the bottom 106 Agilent 1260 Infinity Fraction Collectors User Manual Repairing the Fraction Collector 4 4 Install the new inlet waste tubing assembly into the bolt carrier as shown below Slide in the long ends of the tubings from bottom to top and let the snapper click into position assembly to the ports of the diverter valve IMPORTANT The tubings must not be bent up or downwards The cables must not be twisted View from the bottom 6 Connect the finger tight fittings of the inlet waste tubing 5 The rear tubing with the label with the arrow must be connected to the flow cell of the detector The front tubing must be inserted into the waste container From flow cell To waste 7 A color coded ring on one of the tubings and the valve body indicates which cable belongs to which port IMPORTANT It is absolutely vital to connect these tubings as described in order to maximize their lifetime and operating security Agilent 1260 Infinity Fraction Collectors User Manual 107 4 Repairing the Fraction Collector 8 IMPORTANT The cables must run into the ports of the diverter valve in lines parallel to the horizon
2. Solvent Compartment ao a o Preparative Pump A Column Organizer GPIB or LAN i in E to ChemStation gt 3 Broman E m Pump B E J f CAN Bus UV Vis es F Cable Detector E E 0 rine Autosampler e D e Fraction 2 ee on Collector ma E a OC a Figure 4 Recommended Stack Configuration Fraction Collector in a Preparative Sys tem Rear View If a G1330B ALS thermostat is part of the system it must always be installed underneath the module that is thermostatted injector or Fraction Collector see Installing a Thermostatted Fraction Collector on page 26 The fraction collector should never be positioned on top of a module that generates heat which can lead to an unwanted evaporation of fractions in the fraction collector e g Agilent 1260 Infinity Thermostatted Column Compartment G1316A or Agilent 1260 Infinity Diode Array Detectors Agilent 1260 Infinity Fraction Collectors User Manual 21 1 Installing the Fraction Collector Installing the Fraction Collector Preparation Locate bench space Provide power connections Unpack the Fraction Collector 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 supp
3. According to Runtime Checklist ba F Use Sequence Table Information macro SHUTDOWN MAC go v Katuma pam nRdy Timeout min after loading a new method Fraction Information Fraction Start Locatic Sequence Comment p Figure 29 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 select Run Sequence from the RunControl menu e press F6 74 Agilent 1260 Infinity Fraction Collectors User Manual Configuration and Operation of the Fraction Collector 2 Starting Your Run with Fraction Collection 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 30 1 o Iretrumert Method Sequence Pufy View Abot ae Edit Signal Plot Methods 4 z Seanno Avalabin Syah Seinen Sayas DAD A SepuaimIS4 16 Ramere Lapsed na BABA GignaivSSa 16 Netavencen85 100 i up son Joa mau k CHECXOUTOONE D C aaas oeen E oosa Reedy mom Figure 30 Online Tick Marks Agilent 1260 Infinity Fraction Collectors User Manual 75 2 Configuration
4. contact control Distance 0 0 mm Reserved Locations Plate Type l E E poh Installed Plates 7 Plate 1 front left 7 Plate 2 back left Plate 3 front right T Plate 4 back right Row s Column s Single Location s Figure 15 Use for Sample Recovery This checkbox only appears if multiple fraction collectors are configured see Figure 34 on page 79 Then the last fraction collector can be selected for sample recovery If four fraction collectors are configured the fourth one will automatically used for sample recovery To change the order of your fraction collectors select Configure 1260 Access from Fraction Collector Configuration dialog 2 the Instrument menu In the Configuration dialog box use the Up or Down button to change the configuration order and restart your ChemStation Agilent 1260 Infinity Fraction Collectors User Manual 45 2 46 Configuration and Operation of the Fraction Collector Fraction Delay Volumes The Fraction Delay Volume specifies the volume between the detector cell and fraction collector diverter valve This volume has to be determined during the delay calibration See Delay Volumes and Delay Calibration on page 50 for a detailed description Collection Order The Collection Order describes the way of capillary movement during fraction collection Four different settings are possible e row by row e column by column e shorte
5. Otherwise fraction collector 2 and fraction collector 3 have to be temporarily configured as fraction collector 1 during the delay calibration Before using the preparative scale fraction collector G1364B the delay calibration adapter G1364 87301 has to be removed from the needle carrier assembly 58 Agilent 1260 Infinity Fraction Collectors User Manual Configuration and Operation of the Fraction Collector 2 Performing a Delay Calibration with an MSD 1 Place a vial with the Delay Sensor calibrant G1946 85020 in position 1 of the Autosampler 2 Remove the installed column and connect the capillaries with a zero dead volume connector or a mixer Part No 79835 87330 3 Install the 40 x 2ml vial tray analytical scale or the 15 x 6ml vial tray preparative scale in the fraction collector 4 Connect a bottle of water to Channel A of the main pump 5 Connect a bottle of water with 0 1 acetic acid or ammonium formate to the make up pump 6 Load the method DGCALAS M analytical scale or D CALPS M preparative scale and adjust the flow in the main pump to the flow you will be using for the analysis CAUTION Unlike for UV detectors the delay calibration for the MSD needs to be performed whenever the flow rate is changed 7 Set the flow of the make up pump to the flow rate you will be using for your analysis 8 Set the active splitter to the split ratio as used during operation 9 Save the method to a new name 1
6. __ ____ Figure 19 AFC Delay Volume Calibration Window When you have turned ON the pumps click Continue again to start the delay volume calibration The method to be used for the delay volume calibration is displayed DGCALAS M is the default calibration method for the analytical scale fraction collector and DGCALPS M is the default method for the preparative fraction collector if you want to change to a different method click the Change Method button and select a new method See also Making Your Own Calibration Method on page 64 The Info field shows the current status of the delay volume calibration and gives instructions and warnings 8 Install the delay calibration adapter G1364 87301 by attaching it to the needle carrier assembly G1364B preparative scale only The selected method is loaded and the delay volume calibration sample is injected 9 After the delay calibration has been finished you are prompted to remove the delay calibration adapter G1364B preparative scale only 10 When the calibration run has finished click OK The Calibration Results panel see Figure 20 is displayed Agilent 1260 Infinity Fraction Collectors User Manual Configuration and Operation of the Fraction Collector 2 Delay Volumes and Delay Calibration E Fraction Collector 1 Calibration Results Cale Delay Vol ul GSS sia SS ee DAD1 G13158 DE00900058 Current Signalfs AFCA N Delay senso
7. a Agilent Technologies Notices Agilent Technologies Inc 2010 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 90012 Edition 06 10 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 control Technology
8. 0 5 1 2 4 8 0 0182 0 0364 0 0728 0 146 0 328 0 710 1 49 3 08 6 26 12 6 Table 13 Signal Delay Times for the Agilent 1260 Infinity FLD G1321A Peakwidth min Response time sec Signal delay sec lt 0 005 gt 0 005 gt 0 01 gt 0 03 gt 0 05 gt 0 1 gt 0 2 gt 0 4 lt 0 12 0 12 0 25 0 5 1 2 4 8 0 018 0 073 0 18 0 44 0 96 2 0 4 2 8 6 Agilent 1260 Infinity Fraction Collectors User Manual 53 2 Configuration and Operation of the Fraction Collector CAUTION If the internal signal delay is longer than the delay time tp some of the peak will be lost The maximum allowed signal delay time can be calculated using the following equation Von Signal delay time ma x v Flow rate After calculating the maximum signal delay time a Peakwidth setting can be selected that gives a signal delay time which is shorter than the calculated maximum signal delay time This Peakwidth setting should then be used for the LC purification run 54 We recommend to set the Peakwidth always to gt 0 01 for the DAD and MWD or to gt 0 005 for the VWD If the Peakwidth setting cannot be reduced and the signal delay time is longer than tp it is also possible to increase Vp by adding additional tubing However this higher delay volume will increase the peak dispersion between detector and fraction collector The stop time of the run in the ChemStation must be set to at least To
9. 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 Agilent 1260 Infinity Fraction Collectors User Manual 39 1 40 Installing the Fraction Collector 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 rela
10. Diverter valve 15 Diverter valve to needle 10 Needle 4 Single wavelength absorbance detector working at 654 nm consisting of a LED and a photo diode 3 2 Diverter valve with low internal volume 15 ul switching time lt 100 ms maximum operating pressure 6 bar Agilent 1260 Infinity Fraction Collectors User Manual 145 146 Specifications Table 36 Performance Specifications Agilent 1260 Infinity Analytical scale Fraction Collector G1364C continued Type Specification Cooling Maximum capacity GLP features Interfaces Safety features Optional with additional G1330B performance depending on ambient conditions and the volume of collected fractions 3 fraction collectors in parallel plus one recovery fraction collector connected via 12 Position 13 Port Selector valve PN G1160A Early maintenance feedback EMF electronic records of maintenance and errors 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 Leak detection and safe leak handling error detection and display exhaust fan for fume extraction of hazardous vapors Vials and well plates and capped vials and well plates with closing mats can be used as recom mended by Agilent Technologies see
11. 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 Fraction Collector on top of the thermostat Make sure that the Fraction Collector is correctly engaged in the thermostat locks 10 Remove the plastic cover from the tray base place the air channel adapter 1 into the Fraction Collector 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 Fraction Collector 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 8 Installation of Thermostat and Fraction Collector 11 The analytical scale Fraction Collector is delivered with a pre installed tray compartment divider This divider should only be used with the thermostatted Fraction Collector 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 Fraction Collector are in use Agilent 1260 Infinity Fraction Collectors User Manual 27 1 28 Installing the Fraction Collector Figure 9 Removing the T
12. Transport Unit Assembly Analytical Scale 135 Needle Assemblies 136 Diverter Valve Assembly 137 Tubing Kits 138 Internal Tray Assembly 139 Fraction Collector Accessory Kit 140 s j Agilent Technologies 125 5 Parts and Materials Supported Trays for Fraction Collectors Table 19 Trays for the Fraction Collector Item Description Part Number 1 Full tray for 215 test tubes 12x100 mm G1364 84516 2 Full tray for 126 test tubes 16x100 mm G1364 84525 3 Full tray for 60 round bottom tubes 25x100 mm G1364 84524 4 Full tray for 40 round bottom tubes 30x100 mm G1364 84523 5 Full tray for 4 well plates G1364 84521 6 Std tray for 2 well plates 10 2ml vials G2258 60011 7 Std tray for 2 well plates 10 collecting funnels G1364 84522 8 Funnel for item 7 5022 2200 9 Funnel seal kit for item 7 pack of 10 61364 68730 10 Tubing kit 10T for item 7 pack of 10 G1364 86707 11 Funnel coupler for item 7 G1364 43201 12 Std tray for 100 x 2 ml vials G1313 44510 Std tray for 100 x 2 ml vials thermostattable G1329 60011 13 Half tray for 15 x 6 ml vials G1313 44513 14 Half tray for 40 x 2 ml vials G1313 44512 15 Adapter air channel installed underneath plug channel if G1329 43200 the fraction collector is used with the thermostat 16 Plug channel G1364 47200 17 Half Tray for 40 Funnels G1364 84532 Only one type of well plates can be used at a time in one tray 126 Agilent 1260 Infinity Fraction Collectors User Manual Parts and M
13. automatically be used for recovery 2 Ifthe checkbox Plate 1 and Plate 2 are not selected the 10 funnel position can be used for regular fraction collection With each start of a new sequence the recovery will start at the same positions In order to avoid contamination the vessels that contain the recovery fractions have to be exchanged CAUTION The number of recovery locations automatically defines the maximum number of injections When using the standard tray for two well plates and 10 funnels only ten injections per sequence are executed Semi preparative Operation 80 The analytical scale fraction collector G1364C can be modified for semi preparative operation This allows to use of external collection vessels through funnels at high flow rates above 10 ml min The maximum flow rate depends on the viscosity of the solvent In order to modify the analytical fraction collector the following parts have to be installed e 20 mm needle assembly G1364 87202 e preparative tubing kit G1364 68711 e internal tray preparative scale G1364 63113 The procedures to exchange these parts are described in Replacing the Inlet Waste Tubings on page 105 Replacing the Valve to Needle Tubing on page 109 Exchanging the Analytical Needle Assembly on page 114 and Exchanging the Internal Tray on page 119 In addition the wellplate adapter has to be removed for operation with the shorter needle Agilent 12
14. Collector in the Home Position see Maintenance Functions on page 95 Remove all installed trays from the tray base Position the transport unit of the Fraction Collector in the Change Parts Position Turn OFF the instrument It might be more convenient to remove the needle from its carrier before unscrewing the needle tubing see Maintenance Functions on page 95 1 Using the 4 mm wrench and the 5 16 wrench for counter holding unscrew the valve to needle tubing from the needle Viewed from the bottom 112 Agilent 1260 Infinity Fraction Collectors User Manual Repairing the Fraction Collector 4 2 Holding the needle assembly between your thumb and forefinger slide out the assembly towards the rear of the needle carrier assembly 3 Insert the new needle assembly into the holder of the needle carrier assembly Make sure to push it all the way to the front xi 4 Using the 4 mm wrench and the 5 16 wrench for counter holding connect the valve to needle tubing to the needle Viewed from the bottom On completion of this procedure Re install the needle to the needle carrier assembly if you previously removed it Make sure to slide the needle all the way to the front of the needle carrier assembly clicks into position IMPORTANT After fixing the needle in the needle carrier clip the tubing into the guide on the bottom of the need
15. List of Recommended Vials and Caps on page 129 and List of Recommended Plates and Closing Mats on page 132 Only one type of well plates can be used at a time in one tray Agilent 1260 Infinity Fraction Collectors User Manual Agilent 1260 Infinity Fraction Collectors User Manual A Safety Information Safety Information 148 Lithium Batteries Information 151 Radio Interference 152 Sound Emission 153 Solvent Information 154 Agilent Technologies on Internet 155 j Agilent Technologies 147 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 If you are using flammable solvents cover the well plates with closing mats to avoid the risk o
16. assembly Exchanging the diverter valve Exchanging the internal tray Repairing or exchanging a funnel of the internal tray or funnel tray When worn out when showing visual signs of damage typically once per year When worn out when showing visual signs of damage typically once per year When needle shows indication of damage or blockage When needle shows indication of damage or blockage Or when using the short needle assembly for operation with high test tubes gt 45mm When defective internal external leak valve not switching any more When flow delay sensor defective When defective leaky blocked or contaminated See Replacing the Inlet Waste Tubings on page 105 See Replacing the Valve to Needle Tubing on page 109 See Exchanging the Preparative Needle Assembly on page 112 See Exchanging the Analytical Needle Assembly on page 114 See Exchanging the Diverter Valve on page 116 See Exchanging the Internal Tray on page 119 See Repairing or Exchanging a Funnel of the Internal Tray on page 121 104 Agilent 1260 Infinity Fraction Collectors User Manual Repairing the Fraction Collector 4 Replacing the Inlet Waste Tubings Frequency When contaminated worn out or visibly damaged Typically once every year Tools required None Parts Required Inlet waste tubing assembly in tubing kit preparative scale 0 8 mm ID PN G1364 68711 analytical scale 0
17. be checked Timetable The Timetable can be used to program changes in the Fraction Trigger Mode during the analysis by entering a Time and specifying the trigger settings Trigger Mode Off Peak Based and Time Based can be selected If the Off is selected no fractions are collected The last entry in the timetable has to be the command Off Whenever the Peak Based mode is specified 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 is mandatory if you use Peak Controlled fraction collection but is disabled for Time Based fraction collection When the Time Based mode is chosen two different options are available The of Fractions can be edited to collect a fixed number of equal fractions in a give time interval This time interval is defined by the time value in the current and following timetable line e Timeslices min can be edited to collect fractions with a defined collection time With this option the collection time of the last fraction can be shorter This depends on the overall runtime For editing the Timetable the functions Insert Append Cut Copy and Paste are offered To access the additional sections in the Setup Fraction Collector dialog box click More Time In the time section of the dialog box the Stoptime and the Posttime for the fraction collector can be specified By def
18. 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 Fraction Collectors User Manual In This Guide This manual contains technical reference information about the Agilent 1260 Infinity Fraction Collectors analytical and preparative scale The manual describes the following 1 Installing the Fraction Collector This chapter provides information about the installation of the Agilent 1260 Infinity Fraction Collectors G1364B and C 2 Configuration and Operation of the Fraction Collector This chapter describes the configuration and operation of the fraction collector including guidelines to optimize the system and to avoid problems 3 Troubleshooting and Test Functions This chapter describes the modules built in troubleshooting and test functions 4 Repairing the Fraction Collector This chapter contains instructions on simple repair and maintenance procedures 5 Parts and Materials This chapter contains lists for identification of common repair and maintenance parts 6 Specifications This chapter contains performance specifications of the fraction collectors A Safety Information This appendix provides a safety summary Agilent 1260 Infinity Fraction Collectors User Manual
19. interference limits are still met within the premises 152 Agilent 1260 Infinity Fraction Collectors 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 Fraction Collectors User Manual 153 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 154 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
20. region The plug on the power cord which connects to the rear of the instrument is identical for all types of power cord WARNING 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 WARNING Never use cables other than the ones supplied by Agilent Technologies to ensure proper functionality and compliance with safety or EMC regulations Bench Space The Fraction Collector dimensions and weight see Table 1 on page 12 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 Fraction Collector is installed in a horizontal position The thermostatted Fraction Collector 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 3 1 inches at the rear for electrical connections Ensure the Fraction Collector 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 th
21. required Parts from the accessory kit see Fraction Collector Accessory Kit Contents G1364 68725 on page 16 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 inlet waste tubing assembly and the valve to needle tubing and exchange them if they are worn out or show visible signs of damage WARNING Thoroughly follow the described installation procedures to maximize the lifetime of inlet waste tubing assembly the valve to needle tubing and to avoid potential spills or fraction losses Regularly inspect the tubings and exchange them if they are worn out or show visible signs of damage WARNING This instrument should only be used with solvents that have an ignition temperature higher than 200 C Agilent 1260 Infinity Fraction Collectors User Manual 31 1 Installing the Fraction Collector From flow cell TA 5 7 z A Figure 11 Flow Connections to and from the Fraction Collector Route the waste outlet around the instrument as shown below and into a corrugated waste tubing Fix the waste outlet tubing and the corrugated waste tubing to the instrument with the sti
22. start the hardware initialization process At the end of this process the status LED should be green 21 The Fraction Collector 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 To disconnect the Fraction Collector 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 To avoid personal injury keep fingers away from the needle area during Fraction Collector operation Do not attempt to insert or remove a vial or a plate when the needle is positioned Agilent 1260 Infinity Fraction Collectors User Manual 29 1 Installing the Fraction Collector Installing a Thermostatted Fraction Collector aoaaa Fraction Collector Thermostat cable 0 BBB088 BBB6H6 oO CAN bus AC Power Figure 10 Connections at the Rear of the Thermostatted Fraction Collector 30 Agilent 1260 Infinity Fraction Collectors User Manual Installing the Fraction Collector 1 Flow Connections to the Fraction Collector Preparation Fraction Collector is installed in the LC system Parts
23. the bottom of the right front corner of the instrument Position the transport unit of the Fraction Collector in the Home Position see Maintenance Functions on page 95 Remove all installed trays from the tray base Turn OFF the instrument Agilent 1260 Infinity Fraction Collectors User Manual 119 4 Repairing the Fraction Collector 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 0 A ae Ken a fel 4 Install the new tray by sliding it into position underneath the metal latch 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 1
24. the diverter valve from the 6 Install the new diverter valve to the z arm with its holding screw Plug in the new diverter valve power cable to the valve z arm Agilent 1260 Infinity Fraction Collectors User Manual 117 4 Repairing the Fraction Collector 8 Connect the finger tight fittings of the inlet waste tubing assembly to the ports of the diverter valve IMPORTANT The tubings must not be bent up or downwards The cables must not be twisted View from the bottom 9 A color coded ring on one of the tubings and the valve body indicates which cable belongs to which port IMPORTANT It is absolutely vital to connect these tubings as described in order to maximize their lifetime and operating security 10 IMPORTANT The cables must run into the ports of the diverter valve in lines parallel to the horizon E o On completion of this procedure Re install the tray s in the tray base Start the instrument Close the front cover 118 Agilent 1260 Infinity Fraction Collectors User Manual Repairing the Fraction Collector 4 Exchanging the Internal Tray Frequency When defective Tools required None Parts required Internal tray analytical scale G1364 63113 Internal tray preparative scale G1364 63114 Before beginning this procedure 1 Locate the internal tray assembly with the rinse funnel and flow delay sensor in
25. the well plate the vial or the test tube is removed without removing the complete tray Consequently contaminated fractions unwanted overfill events or split fractions might be observed during the following LC run Assignment of Start Location for Fraction Collection 72 The start location for fraction collection can either be assigned in the Sample Info Figure 28 dialog box in the RunControl menu in the Sequence Parameters Figure 29 dialog box or in the Sequence Table in the Sequence menu All fraction start locations are entered in the following formats Agilent 1260 Infinity Fraction Collectors User Manual Configuration and Operation of the Fraction Collector 2 For vials and test tubes fraction collector vessel number e g 1 Vial 1 For well plates fraction collector plate number row column e g 1 P1 A 01 with fraction collector 1 2 plate number P1 P4 row is A to H and column 1 12 for 96 well plates Note the orientation of the well plate The starting position A1 is always in the back left corner of the plate Sample Info Instrument 1 x Operator Name Bio Chemist r Data File C Prefix Counter Manual Filename IDEFAULT D Subdirectory PROTEOME Path C HPCHEM 1 DATA r Sample Parameters blank run if no entry Fraction Start Location 1 P1 A 01 Sample Name ew Sample Amount a Multiplier fr ISTD Amount fo OOO Dilution ft Figure 28 Sampl
26. vials test tubes of up to 75 mm height well plates and a 40 funnel tray connecting to external locations of any size Agilent 1260 Infinity Fraction Collectors User Manual Installing the Fraction Collector 1 G1364C Thermostatted Fraction Collector analytical scale can be created by additionally ordering and installing a G1330B Fraction Collector Thermostat The terms thermostatted analytical scale Fraction Collector and the preparative scale Fraction Collector refer to these Agilent 1260 Infinity modules in this manual Unless otherwise stated all information in this section is valid for all models Table3 Delivery Checklist for the G1364B preparative scale and G1364C analytical scale Fraction Collector Description Quantity Fraction collector module with inlet waste tubing assembly 1 Power cord local Matching the sockets in your country or region 1 Accessory kit see below 1 Delay Calibrant G1946 85020 1 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 Agilent 1260 Infinity Fraction Collectors User Manual 15 1 16 Installing the Fraction Collector Table5 Fraction Collector Ac
27. 0 Go to Diagnostics menu select Delay volume Calibration from the Fraction Collector Sub menu within the Maintenance menu 11 Press Change method to select the method you created The new method name appears on the info line 12 Start the active splitter 13 Press Continue to execute the Delay Volume Calibration 14 Install the Delay Calibration Adapter G1364 87301 by attaching it to the needle carrier assembly G1364B preparative scale only Agilent 1260 Infinity Fraction Collectors User Manual 59 2 Configuration and Operation of the Fraction Collector The selected method is loaded and the Delay Calibration Sample Injected 15 After the delay calibration has been finished you are prompted to remove the delay calibration adapter G1364B preparative scale only 16 When the Calibration Run is finished press OK 17 Examine the value for the MSD Calculated Delay time 60 Agilent 1260 Infinity Fraction Collectors User Manual Configuration and Operation of the Fraction Collector 2 The delay time should be at least 5 seconds but may in fact be negative see Figure 21 E Fraction Collector 1 Calibration Results Print Close Calibrate Curr Delay Vol ul ee 883 Calc Delay Vol ul DAD1 G13154 US64400562 Calc Delay Time sec 0 294 Detector MSD Curr Delay Time sec Figure 21 Delay Volume Calibration results 18 If the measured delay time is greater than 5 se
28. 2 Transport Unit Assembly Preparative Scale 134 Agilent 1260 Infinity Fraction Collectors User Manual 7 Contents Transport Unit Assembly Analytical Scale 135 Needle Assemblies 136 Diverter Valve Assembly 137 Tubing Kits 138 Internal Tray Assembly 139 Fraction Collector Accessory Kit 140 Specifications 141 Performance Specifications for the Fraction Collectors 142 Safety Information 147 Safety Information 148 General 148 Operation 149 Safety Symbols 150 Lithium Batteries Information 151 Radio Interference 152 Test and Measurement 152 Sound Emission 153 Manufacturer s Declaration 153 Solvent Information 154 Solvents 154 Agilent Technologies on Internet 155 Index 157 Agilent 1260 Infinity Fraction Collectors User Manual Agilent 1260 Infinity Fraction Collectors User Manual 1 Installing the Fraction Collector Site Requirements 10 Unpacking the Fraction Collector 14 Optimizing the Stack Configuration 17 Installing the Fraction Collector 22 Installing a Thermostatted Fraction Collector 26 Flow Connections to the Fraction Collector 31 Fraction Collector Trays 34 Configure Well plate Types 37 Transporting the Fraction Collector 41 f Agilent Technologies 1 Installing the Fraction Collector Site Requirements A suitable site environment is important to ensure optimum performance of the Fraction Collector Power Consideration The Fraction Collector power supply has wide ranging
29. 20 Agilent 1260 Infinity Fraction Collectors User Manual Repairing the Fraction Collector 4 Repairing or Exchanging a Funnel of the Internal Tray Frequency When leaky or contaminated Tools required None Parts required Funnel assembly waste tubing assembly seals for part numbers see Internal Tray Assembly on page 139 Before beginning this procedure 1 Remove the internal tray from the instrument see o Exchanging the Internal Tray on page 119 Position the transport unit of the Fraction E oe Collector in the Home Position see Maintenance Functions on page 95 and remove all installed trays from the tray base Turn OFF the instrument The procedure is almost identical for both the preparative and the analytical scale internal tray but it requires different parts 2 Turn the internal tray upside down and remove the 3 Remove the funnel assembly from the internal tray by funnel s waste tubing through the flow delay sensor screwing it counter clockwise and lifting it out Agilent 1260 Infinity Fraction Collectors User Manual 121 4 Repairing the Fraction Collector 4 Exploded view for the parts of the internal tray Once the funnel has been removed from the internal tray it can be disassembled and defective parts can be replaced funnel screw 1 seals 2 tubing 3 or funnel holder 4 5 Slide the waste tubing of the newly assembled f
30. 25 mm ID PN G1364 68712 or analytical scale 0 15 mm ID G1364 68723 WARNING To avoid personal injury keep fingers away from the needle area during Fraction Collector operation WARNING Follow the described installation procedure exactly to maximize the lifetime of the inlet waste tubings and to avoid potential spills or fraction losses Regularly inspect the tubings and exchange them if they are worn out or show visible signs of damage Agilent 1260 Infinity Fraction Collectors User Manual 105 4 Repairing the Fraction Collector Before beginning this procedure Position the transport unit of the Fraction Collector in the Home Position see Maintenance Functions on page 95 Remove all installed trays from the tray base Position the transport unit of the Fraction Collector in the Change Parts Position see Maintenance Functions on page 95 Turn OFF the instrument Remove the rear end of the Fraction Collector s waste tubing from the waste container unscrew the front end of the Fraction Collector s inlet tubing from the flow cell of the detector 1 Locate the diverter valve with the finger tight fittings of the inlet waste tubing assembly the figure shows the open Fraction Collector seen from the front 2 Unscrew the 2 finger tight fittings of the inlet waste tubing assembly at the diverter valve o R 0
31. 3 Agilent 1260 Infinity Fraction Collectors User Manual Contents 1 Installing the Fraction Collector Site Requirements 10 Power Consideration 10 Power Cords 11 Bench Space 11 Environment 12 Unpacking the Fraction Collector Damaged Packaging 14 Delivery Checklist 14 Accessory Kits 15 Optimizing the Stack Configuration Installing the Fraction Collector 14 17 22 Installing a Thermostatted Fraction Collector 26 Flow Connections to the Fraction Collector 31 Fraction Collector Trays 34 Installing the Fraction Collector Trays 34 Numbering of Vial Test Tube and Well plate Positions 35 Configure Well plate Types 37 Transporting the Fraction Collector 41 2 Configuration and Operation of the Fraction Collector 43 Configuration of the Fraction Collector 44 Configuration of the Fraction Collector in the ChemStation 44 Configuring Multiple Fraction Collectors 48 Agilent 1260 Infinity Fraction Collectors User Manual Contents Delay Volumes and Delay Calibration 50 Delay Times and Volumes 50 Detector Signal Delay 51 Performing a Delay Calibration with an UV Detector 55 Performing a Delay Calibration with an MSD 59 Making Your Own Calibration Method 64 Setting up a Fraction Collector Method 65 Fraction Trigger Mode 66 Fraction Preview 70 Starting Your Run with Fraction Collection 72 Assignment of Start Location for Fraction Collection 72 Online Tick Marks 75 Viewing Your Results 76 Data Analysis 7
32. 6 Report 77 Special Applications 78 Pooling 78 Sample Recovery 79 Semi preparative Operation 80 Using High Test Tubes in the Analytical Scale Fraction Collector 81 Optimizing Fraction Collection 82 Limitations and How to Avoid Problems 83 Check out Procedures 84 Check out Procedure for UV triggered Fraction Collection AS 84 Check out Procedure for UV triggered Fraction Collection PS 87 Application Notes 89 3 Troubleshooting and Test Functions 91 Status Indicators 92 Error Messages 92 Agilent 1260 Infinity Fraction Collectors User Manual Contents Maintenance Functions 92 Transport Unit Self Alignment 92 Status Indicators 93 Power Supply Indicator 93 Instrument Status Indicator 94 Maintenance Functions 95 Transport Unit Self Alignment 97 Step Commands 99 Troubleshooting 100 4 Repairing the Fraction Collector 101 Introduction into Repairing the Fraction Collector 102 Simple Repairs 102 Cleaning the Fraction Collector 102 Overview of Main Repair Procedures 103 Simple Repairs 104 Replacing the Inlet Waste Tubings 105 Replacing the Valve to Needle Tubing 109 Exchanging the Preparative Needle Assembly 112 Exchanging the Analytical Needle Assembly 114 Exchanging the Diverter Valve 116 Exchanging the Internal Tray 119 5 Parts and Materials 125 Supported Trays for Fraction Collectors 126 List of Recommended Test Tubes 128 List of Recommended Vials and Caps 129 List of Recommended Plates and Closing Mats 13
33. 60 Infinity Fraction Collectors User Manual Configuration and Operation of the Fraction Collector 2 In the ChemStation Software the new configuration has to be specified in order to consider the new volumes for the delay calibration In the Configuration dialog box of the fraction collector select More Then change the Inner Capillary Diameter to 0 8 mm as well as the Needle Type to semi prep needle as displayed in Figure 35 on page 81 For further information on delay volumes and delay volume calibration see Delay Volumes and Delay Calibration on page 50 Inner Capillary Needle Type no needle micro needle analytical needle semi prep needle prep needle _ Figure 35 Fraction collector configuration parameters for semi prep operation Using High Test Tubes in the Analytical Scale Fraction Collector In the standard configuration the analytical scale fraction collector G1364C the maximum test tube height is limited to 48 mm This limitation can be extended to 75 mm if e the standard 50 mm needle assembly G1367 87200 is replaced by the 20 mm needle assembly G1364 87202 e the wellplate adapter is removed Agilent 1260 Infinity Fraction Collectors User Manual 81 2 Configuration and Operation of the Fraction Collector Optimizing Fraction Collection Time based Time slices must have a length of at least 0 05 min fraction llecti e Set of Fractions such that length of resulting fractions is at leas
34. 82 3459 130 Agilent 1260 Infinity Fraction Collectors User Manual Parts and Materials 5 Table 26 Screw Caps Caps for Use with the Analytical Scale Fraction Collector only Description Septa 100 Pack Blue polypropylene Clear PTFE red rubber 5182 0717 Green polypropylene Clear PTFE red rubber 5182 0718 Red polypropylene Clear PTFE red rubber 5182 0719 Blue polypropylene Clear PTFE silicone 5182 0720 Green polypropylene Clear PTFE silicone 5182 0721 Red polypropylene Clear PTFE silicone 5182 0722 Agilent 1260 Infinity Fraction Collectors User Manual 131 5 Parts and Materials List of Recommended Plates and Closing Mats Table 27 Recommended Plates and Closing Mats Std Well Plates and Closing Mats for Use with the Analytical Scale Fraction Collector only Description Volume Package Part ml Number 96 polypropylene well plate 0 5 10 5042 1386 96 polypropylene well plate 0 5 120 5042 1385 96 polypropylene deep well pate 1 0 50 5042 6454 96 polypropylene deep well plate with glass inserts caps 0 35 1 5065 4402 and septa pre assembled glass inserts for 5065 44027 0 35 1000 5188 5321 384 polypropylene well plate 0 1 30 5042 1388 96 polypropylene conical well plate 0 18 25 5042 8502 54 x 2ml vial plate 1 5 6 G2255 68700 15 x 6ml vial plate 4 5 1 5022 6539 Plate for 27 self lock Eppendorf tubes 0 5 1 5 2 1 5022 6538 Silicon Closing mats for 96 well plate 50 5042 1389 For use with the analytical sc
35. A B C D and F can t be used Columns 1 Column 1 can t be used 1 2 Columns 1 and2 can t be used 1 4 Columns 1 2 3 and 4 can t be used Agilent 1260 Infinity Fraction Collectors User Manual 47 2 48 Configuration and Operation of the Fraction Collector Table 8 Syntax for the definition of Reserved Locations continued Location Syntax Description 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 Configuring Multiple Fraction Collectors To increase the capacity of the systems up to three fraction collectors can be configured by using the Agilent 1260 Infinity 12 position13 port valve G1160A In addition a fourth fraction collector can be configured for recovery collection The Configuration has to be edited for all fraction collectors The last fraction collector in the configuration can be selected for sample recovery To configure the recovery fraction collector please read Sample Recovery on page 79 e With multiple fraction collectors a fraction start location has to be specified to define the starting position For further details see Assignment of Start Location for Fraction Collection on page 72 Never used the valve settings to define a start location on systems with multiple fraction collectors e We recommend to use inlet tubing of the same length for all fraction collectors Otherwise a Delay Calibration has to be performed fo
36. DADI Threshold Slope 5 00 10 00 10 000 3000 000 mAU Fraction is collected when a peak is detected by I Use MSD for Mass based Fraction Collection C all selected peak detectors at least one selected peak detector Timetable Time Trigger Mode Max Peak Duration min of Fractions Timeslices min Cut Time Rinse Fraction Collection Needle _Auxiliary Max fill volume per location Stoptime aa Eomp E min I At the start of collection as configured zl mi Shee 2 50 i Posttime off A min IV Between fraction collection Store temperature signal I Store delay sensor signal F Store UIB signal Figure 24 Set up Fraction Collector dialog box Agilent 1260 Infinity Fraction Collectors User Manual 65 2 66 Configuration and Operation of the Fraction Collector Fraction Trigger Mode Use Timetable Enables the Timetable Peak based If Peak based is selected the collection of a fraction is triggered by the signal of the detector e g the Agilent 1260 Infinity Diode Array Detector or variable wavelength detector The detailed trigger conditions are specified in the Peak Detectors table In the peak based trigger mode all entries in the Timetable are ignored Max Peak Duration Defines a maximum collection time in case that the signal does not reach the condition to cut the fraction as exhibited in Figure 25 The could be caused by tailing peaks or if the baseline is drifting during gradient runs The default va
37. E 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 Fraction Collectors User Manual Repairing the Fraction Collector 4 Replacing the Valve to Needle Tubing Frequency When contaminated worn out or visibly damaged Typically once every year Tools required Wrench open end 4mm PN 8710 1534 supplied in accessory kit Wrench open end 1 4 5 16 inch PN 8710 0510 supplied in accessory kit Parts Required Valve to needle tubing assembly included in tubing kit preparative scale 0 8 mm ID PN G1364 68711 analytical scale 0 25 mm ID PN G1364 68712 or analytical scale 0 15 mm ID G1364 68723 WARNING To avoid personal injury keep fingers away from the needle area during Fraction Collector operation WARNING Follow the described installation procedure exactly to maximize the lifetime of the valve to needle tubing and to avoid potential spills or fraction losses Regularly inspect the tubings and exchange them if they are worn out or show visible signs of damage Before beginning this procedure 1 Locate the diverter valve with the finger tight fittings of the valve to needle tubing assembly the figure shows the open Fraction Collector seen from the front Position the transport unit of the Fraction Collector in the Home Position see Maintenance Functions on page 95 Remove all inst
38. 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 WARNING notice denotes a
39. 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 2ml vials or 10 funnels std trays Plate in the front position P1 Plate in the back position P2 Vessel Al A2 B1 B2 Vials funnels 1 10 With 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 Agilent 1260 Infinity Fraction Collectors User Manual 35 1 Installing the Fraction Collector With the 40 60 125 or 215 position test tube full trays Numbering starts in front left corner in columns to the back and then to the right WARNING If you are using flammable solvents remove the plates from the Fraction Collector after turning it OFF You avoid the risk of building explosive gas mixtures in the instrument WARNING If you are using flammable solvents cover the plates You avoid the risk of building explosive gas mixtures in the instrument WARNING Closing mats with adhesive can give some contamination in the system The adhesive is soluble in most of the solvents used in HPLC WARNING In general do not use closing mats with
40. User Manual 69 2 70 Configuration and Operation of the Fraction Collector Fraction Preview To determine the appropriate fraction collection parameters the Agilent ChemStation provides a valuable tool that becomes accessible by pushing the button labelled Fraction Preview Tool Figure 26 in the Peak Detectors section Fraction preview peakeasea x p Peak Detectors Detector Working Mode Up Slope Unit s Down Slope Unit s Threshold Unit Upper Threshold Unit Unit 1 DADI _ Threshold Slope 5 00 5 00 5 000 I 3000 000 mau 2 _ uB Off 5 00 5 00 5 000 3000 000 mv m Timetable Insert Time Trigger Mode Max Peak Duration min of Fractions Timeslices min Append Paste Refresh r Fraction Preview DAD1 A Sig 254 4 Ref 550 100 DEMOW05 0101 D zeje mau i o 1 2 3 4 5 6 min Threshold LoadSignal OK Cancel Help Figure 26 Fraction Preview dialog box The Fraction Preview screen allows to test the fraction collection parameters against an example 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 To load a chromatogram e g a pilot run click Load Signal Parameters can now be changed either manually in the Detector Table and Timetable or graphically in
41. adapter G1364 87301 has to be removed from the needle carrier assembly 64 Making Your Own Calibration Method You make your own calibration method by editing one of the default calibration methods 1 Load the default calibration method DGCALAS M in the case of an analytical system and DGCALPS M in the case of a preparative system 2 Change the method parameters flow runtime solvent composition injection volume or detector parameters to suit your adapted calibration procedure 3 Save the method with a new name in the method folder for your instrument 4 Follow the appropriate procedure as described in the previous sections for running the method Agilent 1260 Infinity Fraction Collectors User Manual Configuration and Operation of the Fraction Collector 2 Setting up a Fraction Collector Method In order to setup or change the method parameters of your fraction collector 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 24 In the Setup Fraction Collector dialog box general method settings are specified Set up Fraction Collector LC113 x Fraction Trigger Mode C Off C Use Timetable Max Peak Duration 0 5 min Peak Detectors Detector Working Mode Up Slope Unit s Down Slope Unit s Threshold Unit Upper Threshold Unit Unit 1
42. adhesive The Fraction Collector has no prepunch needle therefore the adhesive will clog the needle after several injections 36 Agilent 1260 Infinity Fraction Collectors User Manual Installing the Fraction Collector 1 Configure Well plate Types If the plate you are using is not found on the List of Recommended Plates and Closing Mats on page 132 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 OOS OO gt TO ie nd a ae Gn es CC OO 2 ae ae Co column Figure 13 Well plate Dimensions straight Agilent 1260 Infinity Fraction Collectors User Manual 37 1 38 Installing the Fraction Collector Configure Well plate Types Figure 14 Well plate Dimensions staggered Agilent 1260 Infinity Fraction Collectors User Manual Installing the Fraction Collector 1 Table7 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
43. age 19 show the configuration recommended for the Fraction Collector in an analytical scale system Figure 3 on page 20 and Figure 4 on page 21 show the configuration recommended for the Fraction Collector with a preparative scale system Agilent 1260 Infinity LC Purification Systems can comprise many different LC modules This section describes some common stack configurations and general considerations On more complex systems e g with two or more fraction collectors or with mass selective detector other stack configurations might show advantages Agilent 1260 Infinity Fraction Collectors User Manual 17 Installing the Fraction Collector Optimizing the Stack Configuration Solvent Compartment Degasser Pump l Autosampler il i ris etector i i Eas a Fraction Column E Collector Compartment Figure 1 Recommended Stack Configuration Fraction Collector in an Analytical Sys tem Front View NOTE For information about connecting an Agilent LC MSD a Non 1260 Infinity or Non UV Vis detector to the system please refer to the User s Guide for Purification High Throughput System PN G2262 90010 or to the Installation Note for the Mass Based Fraction Collection Kit 18 Agilent 1260 Infinity Fraction Collectors User Manual Installing the Fraction Collector 1 Solvent Compartment Degasser Pump GPIB or LAN eg pe to LC ChemStation a UV Vis Autosampler D
44. ale Fraction Collector only Only one type of well plates can be used at a time in one tray If you are using flammable solvents remove the plates from the Fraction Collector after turning it OFF You avoid the risk of building explosive gas mixtures in the instrument 132 Agilent 1260 Infinity Fraction Collectors User Manual Parts and Materials 5 WARNING If you are using flammable solvents cover the plates This action avoids the risk of building explosive gas mixtures in the instrument WARNING Closing mats with adhesive can give some contamination in the system The adhesive is soluble in most of the solvents used in HPLC WARNING In general do not use closing mats with adhesive The Fraction Collector has no prepunch needle therefore the adhesive will clog the needle after several injections Agilent 1260 Infinity Fraction Collectors User Manual 133 5 Parts and Materials Transport Unit Assembly Preparative Scale Table 28 Transport Unit Assembly Preparative Scale Item Description Part Number 1 Transport Unit Assembly Preparative Scale G1364 60018 includes items 2 and 4 2a b Tubing diverter valve to needle 0 8 mm ID G1364 86711 3 Diverter valve assembly G1364 61901 4 Needle carrier assembly preparative scale G1364 60011 5 Needle assembly preparative scale G1364 87201 6 Delay Calibration Adapter not shown 61364 87301 Figure 42 Transport Unit Assemb
45. alled trays from the tray base Position the transport unit of the Fraction Collector in the Change Parts Position see Maintenance Functions on page 95 and turn off the instrument It might be more convenient to remove the needle from its carrier before unscrewing the needle tubing Agilent 1260 Infinity Fraction Collectors User Manual 109 4 Repairing the Fraction Collector 2 Unscrew the finger tight fitting of the valve to needle tubing assembly at the diverter valve o R 0 3 Using the 4 mm wrench and the 5 16 wrench for counter holding unscrew the valve to needle tubing from 4 Un clip the tubing from the bottom of the needle carrier assembly 5 Slide the tubing through the hole in the needle carrier assembly from bottom to top and out of the holder in the z arm assembly 110 Agilent 1260 Infinity Fraction Collectors User Manual Repairing the Fraction Collector 4 6 Install the new valve to needle tubing assembly by clipping it in to the holder in the z arm assembly 1 Important and slide it through the hole in the z arm 2 and out on the bottom of the needle carrier assembly top to bottom Screw the finger tight fitting into the port of the diverter valve 3 7 Using the 4 mm wrench and the 5 16
46. and Operation of the Fraction Collector Viewing Your Results Viewing Your Results Data Analysis 76 In order to display the tick marks for the collected fractions on the screen click Signal options from the Graphics menu Then choose Separated in the Layout dropdown list To review your chromatograms file information and a fraction list select the Data Analysis view and press Fraction Task as displayed in Figure 31 Purification Task Chromatogram with Fraction Tick Marks F instrument 1 Fie Graphics Integration Calibration Report S SCREENS CHECKOUTOO0001 D pif Am ISTD Amount Multiplier Dilution Data File jo i 1 CHECKOUTO Method Name CHECKOUTAS M Date Time Operator p 1 3 2006 2 31 28 fl File Information i File Path C CHEM32 1 DATA SCREENS a Fraction Fraction Location Start Time min End Time min _ Volume pl Reason Date 03 Jan 06 15 31 28 E 1 P1A 01 1 75 1 32 187 71 DAD1 Sample 2 1 P1 4 02 214 227 148 33 DAD1 Sample Into 3 1 P1 4 03 3 25 342 188 54 DAD1 Barcode 4 1 P 14 04 4 87 5 12 268 33 DAD1 Dperator Method CHECKOUTAS M Analysis Time 7 133 min Sampling Rate 0 0033 min 0 198 sec 2141 datapoints gt Ready 0 0033 min 0 198 sec 2141 datapoints Fraction table Figure 31 Fraction Task bu
47. anized Clear glass 2 5182 0544 5183 4504 5183 4507 Clear glass 2 5182 0546 5183 4505 5183 4508 write on spot Amber glass 2 5182 0545 5183 4506 5183 4509 write on spot Table 23 Screw Top Vials Caps for Use with the Analytical Scale Fraction Collector only Description Volume ml 100 Pack 1000 Pack 100 Pack silanized Clear glass 2 5182 0714 5183 2067 5183 2070 Agilent 1260 Infinity Fraction Collectors User Manual 129 5 Parts and Materials Table 23 Screw Top Vials Caps for Use with the Analytical Scale Fraction Collector only continued Description Volume ml 100 Pack 1000 Pack 100 Pack silanized Clear glass 2 5182 0715 5183 2068 5183 2071 write on spot Amber glass 2 5182 0716 5183 2069 5183 2072 write on spot Table 24 Crimp Caps Caps for Use with the Analytical Scale Fraction Collector only Description Septa 100 Pack Silver aluminum Clear PTFE red rubber 5181 1210 Silver aluminum Clear PTFE red rubber 5183 4498 1000 Pack Blue aluminum Clear PTFE red rubber 5181 1215 Green aluminum Clear PTFE red rubber 5181 1216 Red aluminum Clear PTFE red rubber 5181 1217 Table 25 Snap Caps Caps for Use with the Analytical Scale Fraction Collector only Description Septa 100 Pack Clear polypropylene Clear PTFE red rubber 5182 0550 Blue polypropylene Clear PTFE red rubber 5182 3458 Green polypropylene Clear PTFE red rubber 5182 3457 Red polypropylene Clear PTFE red rubber 51
48. ased 68 timetable 66 68 transport 41 transport unit assembly 41 134 135 analytical scale 135 preparative scale 134 transport unit self alignment 92 97 transporting the fraction collector 41 tray combinations 35 tray compartment divider 28 tray height adjustment 24 trays 44 126 142 145 installing 33 numbering of vessel positions 35 trigger modes 142 144 troubleshooting and test functions 91 tube height 44 tube volume 44 tubes 128 tubing kits 138 U UIB 67 universal interface box 67 unpacking the fraction collector 14 upper threshold 67 V valve to needle tubings 109 vial contents temperature 142 vials and caps 129 W waste tubings 33 weight 11 well plates 35 47 configuration 47 working mode 67 Agilent 1260 Infinity Fraction Collectors User Manual Index 159 Index 160 Agilent 1260 Infinity Fraction Collectors User Manual www agilent com In This Book This manual contains technical reference information about the Agilent 1260 Infinity Fraction Collectors The manual describes the following e configuration and operation of the fraction collector e troubleshooting and test functions e simple repairs e parts and materials e specifications e safety information Agilent Technologies 2010 Printed in Germany 06 10 G1364 90012 ee Agilent Technologies
49. assembly analytical scale includes items 2 7 G1364 63114 Internal tray assembly preparative scale includes items2 7 G1364 63113 2 Screw seat adapter 5022 2200 3 Seal funnel reorder No pack of 10 G1364 68730 4a Waste Tubing Kit 0 5T analytical scale 61364 86708 4b Waste Tubing Kit 0 8T preparative scale G1364 86719 5 Funnel coupler G1364 43201 6 Tray internal Order Item 1 7 Flow delay sensor not shown Order Item 1 9 6 3 4 a b db aB Z 2v A q rA ae Figure 47 Internal Tray Assembly Analytical Scale Agilent 1260 Infinity Fraction Collectors User Manual 139 5 Parts and Materials Fraction Collector Accessory Kit Table 34 Fraction Collector Accessory Kit Contents G1364 68725 Description Quantity Part Number Wrench open end 4mm 1 8710 1534 Wrench open end 1 4 5 16 inch 1 8710 0510 Hex key 2 0 mm 1 8710 2476 Finger tight fittings 1 16 f 120 3 0100 1516 Waste tubing 1 2 m 1 5062 2463 Y Type Fitting for waste tubing 1 5065 9971 CAN cable 1 m 1 5181 1519 Air channel adapter 1 61329 43200 Sticking clamp for corrugated waste tubing large 3 5065 9976 Sticking clamp for waste tubing small t 3 5065 9976 ESD wrist strap 1 9300 1408 Tray for 15 x 6 ml vials 1 61313 44513 Tray for 40 x 2 ml vials 1 G1313 44512 Reorder gives pack of 2 t Reorder gives 5m Reorder gives pack of 10 Reorder gives 10 clamps for corrugated waste tub
50. aterials 5 SSS lt a ae ee Pig SS SES S SS SSE Sy CSP SS E E oeae See Se ices se OS PESTS The items in this figure are displayed in different scaling factors Figure 41 Trays Agilent 1260 Infinity Fraction Collectors User Manual 127 5 Parts and Materials List of Recommended Test Tubes Table 20 Round Bottom Test Tubes outer diameter height Volume recommended tray type Part Number 30 mm 100 mm 45 ml G1364 84503 40 tubes 5042 6459 100 pk 30 mm 48 mm 20 ml G1364 84503 40 tubes 5042 6458 100 pk 25 mm 100 mm 35 ml G1364 84504 60 tubes 5042 6470 100 pk 16 mm 100 mm 19 ml G1364 84505 126 tubes 5022 6532 250 pk 16 mm 48 mm 9 ml G1364 84505 126 tubes 5022 6533 100 pk 12 mm 100 mm 11 ml G1364 84506 215 tubes 5022 6431 250 pk 12 mm 48 mm 5 ml G1364 84506 215 tubes 5022 6435 100 pk 128 Agilent 1260 Infinity Fraction Collectors User Manual Parts and Materials 5 List of Recommended Vials and Caps Table 21 Crimp Top Vials Caps for Use with the Analytical Scale Fraction Collector only Description Volume ml 100 Pack 1000 Pack 100 Pack silanized Clear glass 2 5181 3375 5183 4491 Clear glass 2 5182 0543 5183 4492 5183 4494 write on spot Amber glass 2 5182 3376 5183 4493 5183 4495 write on spot Table 22 SnapTop Vials Caps for Use with the Analytical Scale Fraction Collector only Description Volume ml 100 Pack 1000 Pack 100 Pack sil
51. ault the Stoptime is set to as pump and the posttime is switched OFF Rinse Fraction Collection Needle Analytical Scale only If Discrete Fractions is selected as Collection Mode see also Collection Mode on page 46 you can setup a needle rinse step before the fraction collection and or between fractions Then the needle will move to the funnel Agilent 1260 Infinity Fraction Collectors User Manual Configuration and Operation of the Fraction Collector 2 on the internal tray and the diverter valve will switch to flush the needle in order to avoid carry over from the previous fraction The instrument will determine if it is possible to rinse the needle before the next fraction is expected If you have recovery positions in your fraction collector or if you are using one fraction collector for sample recovery in a multiple fraction collector configuration the function rinse between fractions is ignored 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 Additional check boxes in this section provide the opportunity to Store the temperature signal and the UIB signal Agilent 1260 Infinity Fraction Collectors
52. capability see Table 1 on page 12 Consequently there is no voltage selector in the rear of the Fraction Collector There are also no externally accessible fuses as automatic electronic fuses are integrated in the power supply The thermostatted fraction collector comprises two modules the fraction collector G1364B or G1364C 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 control cable and both are turned on by the Fraction Collector module The thermostat power supply has two externally accessible fuses To disconnect the Fraction Collector 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 Fraction Collector from line power unplug the power cord from the Fraction Collector and the thermostat The power supplies still use some power even 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 10 Agilent 1260 Infinity Fraction Collectors User Manual Installing the Fraction Collector 1 Power Cords Your Fraction Collector is delivered with a power cord which matches the wall socket of your particular country or
53. capacities and trays Maximum tube plate height Maximum tube volume Maximum flow rate Delay volumes ul Delay calibration sensor Diverter valve 4x well plates full tray MTP 2x well plates std tray 10 funnels with external containers 1 half tray e 2x well plates std tray MTP 10 x 2 ml vials 1 half tray 100 x2 ml in std tray 1 half tray 3x40x2 ml in half tray 3x 40 funnels in half tray e 3x15x6mlin half tray e Full tray with 40 test tubes 30 mm OD max height 48 mm 20 ml vol e Full tray with 60 test tubes 25 mm OD max height 48 mm e Full tray with 126 test tubes 16 mm OD max height 48 mm Full tray with 215 test tubes 12 mm OD max height 48 mm Installed trays are automatically detected and identified Installed plates and vials can be detected when operating in the needle into location mode max height can be extended by using the short needle assembly G1364 87202 48 mm with long needle assembly G1367 87200 75 mm with short needle assembly G1364 87202 ca 20 ml with 48 mm test tubes ca 30 ml with 75 mm test tubes or unlimited if funnels are used with external containers 10 ml min depending on viscosity and generated back pressure max 6 bar at the diverter valve The analytical scale fraction collector can be modified for flow rates gt 10 ml min Fraction collector inlet to diverter valve 50 typical depends on the length of the tubing
54. cessory Kit Contents G1364 68725 Description Quantity Part Number Wrench open end 4mm 1 8710 1534 Wrench open end 1 4 5 16 inch 1 8710 0510 Hex key 2 0 mm 1 8710 2476 Finger tight fittings 1 16 f 120 3 0100 1516 Waste tubing 1 2 m 1 5062 2463 Y Type Fitting for waste tubing 1 5065 9971 CAN cable 1 m 1 5181 1519 Air channel adapter 1 G1329 43200 Sticking clamp for corrugated waste tubing large 3 5065 9976 Sticking clamp for waste tubing small 3 5065 9976 ESD wrist strap 1 9300 1408 Tray for 15 x 6 ml vials 1 61313 44513 Tray for 40 x 2 ml vials 1 G1313 44512 Reorder gives pack of 2 t Reorder gives 5m Reorder gives pack of 10 Reorder gives 10 clamps for corrugated waste tubing large tt Reorder gives 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 Fraction Collector 1 G1330 81600 Reorder gives 5 m Agilent 1260 Infinity Fraction Collectors User Manual Installing the Fraction Collector 1 Optimizing the Stack Configuration If your Fraction Collector is part of a system you can ensure optimum performance and minimum delay volume by installing the following configuration Figure 1 and Figure 2 on p
55. check out procedures for the UV based trigger are described For the mass based system refer to the Agilent 1260 Infinity Purification System User Manual G2262 90011 Check out Procedure for UV triggered Fraction Collection AS Prerequisites V The Agilent 1260 Infinity LC System has been flushed thoroughly with the liquid phase as described in Table 14 and the lamp s of the UV detector have been switched on for at least 15 minutes to provide a stable baseline V The Agilent 1260 Infinity Fraction Collector has been installed and configured The delay volume has been calibrated as described in Performing a Delay Calibration with an UV Detector on page 55 V The 40 vial tray G1313 44512 with at least four 2ml vials is installed 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 40 vial tray Then click on Yes in displayed dialog box 84 Agilent 1260 Infinity Fraction Collectors User Manual Configuration and Operation of the Fraction Collector 2 Table 14 Method Parameters for UV triggered Fraction Collection Check out AS Agilent 1260 Infinity modules Column Sample Flow Stoptime Solvent A Solvent B Wavelength VWD Wavelength DAD MWD Peakwidth Responsetime Injection volume Column temperature Fraction trigger mode Max peak duration DAD MWD working mode Upslope Downslope Threshold Upper threshold e standard or micro degass
56. cking holders part of the G1364 68725 accessory kit as shown below Route the corrugated waste tubing into a waste container 32 Agilent 1260 Infinity Fraction Collectors User Manual Installing the Fraction Collector Flow Connections to the Fraction Collector Figure 12 Fixing and Routing the Waste Tubings Agilent 1260 Infinity Fraction Collectors User Manual 1 33 1 Installing the Fraction Collector Fraction Collector 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 3 Adjust the top plate of the test tube tray for the correct tube height if required 4 Load the Fraction Collector tray with Fraction Collector well plates test tubes or vials as required 5 Slide the Fraction Collector tray into the Fraction Collector so that the rear of the tray is seated firmly against the rear of the tray area Installed trays are automatically detected and identified 6 Press the front of the Fraction Collector tray down to secure the tray in the Fraction Collector 7 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 34 Agilent 1260 Infinity Fraction Collectors User Manual Installing the Fraction Collector 1 Numbering of Vial Test Tube and Well plate
57. conds you can skip to step 23 otherwise continue with step 19 CAUTION If the measured delay time is significantly greater than 5 seconds you should continue with the calibration at step 16 to avoid losing fractions at the end of the run 19 Determine how much additional time is needed to create a 5 second delay For example if the time displayed is 1 sec a six second delay is needed 20 Using the main pump flow rate determine the desired additional delay volume For example at 3 ml min and a six second delay an additional 3 0 x 6 60 or 0 3 ml is required 21 Cut off a sufficient piece of the supplied ETFE tubing G1968 60500 to provide the needed volume The tubing s volume is 8 1 ul cm In this case 37 cm would be required 22 Add the tubing between the splitter and the fraction collector or if multiple fraction collectors are installed between the splitter and the G1160A valve 23 Perform the calibration again Verify the time is sufficient gt 5 seconds see Figure 22 Agilent 1260 Infinity Fraction Collectors User Manual 61 2 Configuration and Operation of the Fraction Collector Delay Volumes and Delay Calibration E Fraction Collector 1 Calibration Results Print Close Calibrate Detector Curr Delay Vol al Calc Delay Vol ul DAD1 G13154 US64400562 937 1 Cur Delay Time sec Calc Delay Time sec 11 991 MSD Figure 22 Delay time greater than 5 seconds 24 Press Calibrate if you wa
58. e Info dialog box All settings made in the Sequence Table overrule the settings made 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 and Next Location In the former case fraction collection is started at the next free plate and in the latter case fraction collection is started at the next free location If a fraction start location is specified the collector will start at this location even if this vessel has been used during a previous run Agilent 1260 Infinity Fraction Collectors User Manual 73 2 Configuration and Operation of the Fraction Collector If no fraction start location has been specified the fraction collection starts at the first empty vessel or location CAUTION With multiple fraction collectors a fraction start location must be specified e g 1 P1 A 01 or 2 Vial 10 Otherwise the fraction collection of the next run will start at the fraction collector where the last run stopped Never used the valve settings to define a start location Sequence Parameters Instrument 1 L x Operator Name Bio Chemist Data File Bar Code Reader Auto Prefix Counter F Use In Sequence Prefix Counter On a bar code mismatch slat cout C Inject anyway Subdirectory PROTEOME Don t inject Path C HPCHEM 1 DATA Part of methods to run m Shutdown V Post Sequence Cmd Macro
59. e failure a list of probable causes of the problem and a list of suggested actions refer to the Agilent 1260 Infinity Fraction Collectors Service Manual G1364 90111 Maintenance Functions The maintenance functions position the transport unit and needle carrier assembly on certain positions for maintenance homing parking or delay calibration see Maintenance Functions on page 95 Transport Unit Self Alignment The transport unit alignment with the transport unit and the well plate tray is required to compensate for larger deviations in positioning the needle carrier assembly The transport unit self alignment is required after disassembling the system or when you exchange the transport unit the needle carrier assembly or the MTP main board This function is in the Diagnose screen of the ChemStation or the Agilent Instant Pilot G4208A The sample transport self alignment requires one of the 4 well plate trays G1364 84501 but well plates MUST NOT be installed 92 Agilent 1260 Infinity Fraction Collectors User Manual Troubleshooting and Test Functions 3 Status Indicators Two status indicators are located on the front of the Fraction Collector The lower left indicates the power supply status the upper right indicates the Fraction Collector status Status indicator green yellow red Line power switch with green light Figure 37 Location of Status Indicators Power Supply Indicator The powe
60. e thermostatted Fraction Collector it is recommended to position the modules in two stacks see Recommended Stack Configuration Fraction Collector in an Analytical System Front View on page 18 Make sure that in this configuration there is 25 cm 10 inches space on either side of the thermostatted Fraction Collector for the circulation of air Agilent 1260 Infinity Fraction Collectors User Manual 11 1 Installing the Fraction Collector Environment Your Fraction Collector will work within specifications at ambient temperatures and relative humidity as described in Table 1 and Table 2 on page 13 CAUTION Do not store ship or use your Fraction Collector under conditions where temperature fluctuations may cause condensation within the Fraction Collector Condensation will damage the system electronics If your Fraction Collector was shipped in cold weather leave it in its box and allow it to warm up slowly to room temperature to avoid condensation Table1 Physical Specifications Fraction Collectors G1364B G1364C 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 Line frequency Power consumption apparent power Power consumption active power 100 240 VAC 10 Wide ranging capability 50 or 60 Hz 5 200 VA Maximum 180 W Maximum Ambient operating temperature Ambient non operating temp
61. ed fraction collection the system delay times tp and tpg can be calculated by dividing the delay volumes by the flow rate v Diverter valve Waste Figure 16 Delay Volumes and delay times The delay volume Vp is a system parameter it depends on the installed fraction collector tubing and the fraction collector needle It is approximately 23 ul for the fraction collector AS and approximately 120 ul for the fraction collector PS Delay volume Vp which is specified in the Fraction Collector Configuration window is determined using the Delay Volume Calibration feature of the ChemStation software When a peak is detected during a purification run Figure 17 the diverter valve is triggered using the following delay time calculations Start of fraction collection t t tp End of fraction collection t tg tp tpg Agilent 1260 Infinity Fraction Collectors User Manual Configuration and Operation of the Fraction Collector 2 0 1 2 3 Figure 17 Chromatogram with peak start t0 and peak end tE Detector Signal Delay 8 g min Every Agilent 1260 Infinity detector that is used for triggering fractions has an internal signal delay caused by filtering the raw data The signal delay depends on the Peakwidth setting of the detector and is accounted for when the diverter valve is triggered Table 9 and Table 11 list the internal signal delay times for different Peakwidth settings Table9 Signal delay times for the Ag
62. en fraction collection is ignored Needle The option into location under Needle Movement in the fraction collector Movement configuration must only be used for capped 2 or 5 ml vials or well plates Using other or open vials with this command can lead to a Movement failed error Replacing When replacing filled tubes vials or well plates from the fraction collector fraction make sure to remove and re insert the complete tray Otherwise the fraction containers collector will not recognize that the fraction containers were emptied Pooling When pooling fractions overfill protection no longer exists It is the user s responsibility to make sure that all fraction collection locations are large enough to completely collect all pooled fractions If a fraction collection location is overfilled an error message occurs and the pump is shut OFF Agilent 1260 Infinity Fraction Collectors User Manual 83 2 Configuration and Operation of the Fraction Collector Check out Procedures The purpose of the check out injection is the verification of the correct installation and configuration of the Agilent 1260 Infinity Purification System In addition the checkout procedure can used for troubleshooting if the Agilent 1260 Infinity Fraction Collector doesn t operate as expected The checkout procedure depends on the flow range analytical scale AS or preparative scale PS and on the peak trigger UV based or mass based In the following sections the
63. er isocratic quaternary or binary pump e autosampler well plate sampler column compartment e diode array detector multi wavelength detector or variable wavelength detector with analytical flow cell e fraction collector analytical scale ZORBAX Eclipse XDB C18 5 um 150 x 4 6mm 993967 906 or equivalent Check out sample 01080 68704 0 8 ml min 6 minutes 10 HPLC grade water 90 HPLC grade acetonitrile 254 nm Signal A 254 16nm Reference 360 80nm gt 0 1min 2s Tul 40 0 C Peak based 0 5 min Threshold slope 5 mAU s 10 mAU s 10 mAU 3000 mAU Agilent 1260 Infinity Fraction Collectors User Manual 85 2 Configuration and Operation of the Fraction Collector Check out Procedures Expected Result Run a single injection under the conditions as described in Table 14 You should observe a chromatogram with four peaks as displayed in Figure 36 on page 86 Four fractions should have been collected in the fraction collector vials The retention times of the peaks could be different depending on the system configuration e g flow cell type capillary volumes type and age of column E Instrument 1 ax 1 Purify View Abort CHECKOUTAS M SINGLERUNS Date Time Operator vial Sample Name Method Name Sample Am ISTD Amount Multiplier Dilut
64. erature Humidity Operating Altitude Non operating altitude Safety standards IEC CSA UL 4 55 C 41 131 F 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 Fraction Collector Installation Category II Pollution Degree 2 For indoor use only 12 Agilent 1260 Infinity Fraction Collectors User Manual Table 2 Physical Specifications Thermostat G1330B Installing the Fraction Collector 1 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 Line frequency Power consumption active power Power consumption apparent power 100 120 or 220 240 VAC 10 50 or 60 Hz 5 210 W 260 VA Wide ranging capability Maximum Maximum Ambient operating temperature Ambient non operating temperature Humidity Operating Altitude Non operating altitude Safety standards IEC CSA UL EN 4 40 C 41 131 F 40 70 C 4 158 F lt 95 at 25 40 C 77 104 F Up to 2000 m 6500 ft Up to 4600 m 14950 ft Installation Category II Pollution Degree 2 For indoor use only Non condensing For storing the Fraction Collector Agilent 1260 Infinity Fraction Collectors User Manual 13 1 Installing the Fraction Collector Unpacking the Fraction Col
65. etector CAN Bus cable Fraction Collector Column Compartment Figure 2 Recommended Stack Configuration Fraction Collector in an Analytical Sys tem Rear View If a G1330B ALS thermostat is part of the system it must always be installed underneath the thermostatted injector or Fraction Collector see Installing a Thermostatted Fraction Collector on page 26 The fraction collector should never be positioned on top of a module that generates heat This could lead to an unwanted evaporation of fractions in the fraction collector e g Agilent 1260 Infinity Thermostatted Column Compartment G1316A or Agilent 1260 Infinity Diode Array Detectors Agilent 1260 Infinity Fraction Collectors User Manual 1 Installing the Fraction Collector Optimizing the Stack Configuration Solvent Compartment Preparative Pump A Column Organizer Preparative Pump B UV Vis Detector Autosampler Fraction Collector Figure 3 Recommended Stack Configuration Fraction Collector in a Preparative Sys tem Front View NOTE For information about connecting an Agilent LC MSD a Non 1260 Infinity or Non UV Vis detector to the system please refer to the User s Guide for Purification High Throughput System PN G2262 90010 or to the Installation Note for the Mass Based Fraction Collection Kit 20 Agilent 1260 Infinity Fraction Collectors User Manual Installing the Fraction Collector 1
66. f building explosive gas mixtures WARNING After a leak in the sampler make sure the leak plane is cleaned and dry 148 Agilent 1260 Infinity Fraction Collectors 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 haza
67. fications 141 142 analytical scale 144 preparative scale 142 physical specifications 12 fraction collector 12 thermostat 13 plates 132 pooling 78 83 power consideration 10 power cords 11 power requirements 10 power supply indicator 93 preparative needle assembly 112 preview 70 recommended tubes 128 recommended vials and caps 129 recovery 79 recovery fraction collector 49 removing tray compartment divider 28 repairing the fraction collector 101 repairs analytical needle assembly 114 cleaning the fraction collector 102 funnel 121 inlet waste tubings 105 internal tray 119 main procedures overview 103 needle 112 114 preparative needle assembly 112 simple repairs 102 104 valve to needle tubings 109 report 77 reserved locations 47 reset fill volumes 72 rinsing 68 83 S safety features 146 safety information on lithium batteries 151 sample recovery 45 79 semi preparative operation 80 sheet metal kit 140 shipping 41 simple repairs 102 104 site requirements 10 slope only 67 Specifications 141 specifications 12 141 142 stack configuration 19 21 rear view 19 21 start location 72 start run 74 start sequence 74 status indicators 92 93 storage 12 T temperature 12 142 test tube height 81 test tubes 81 128 thermostat electrical connections 30 threshold Agilent 1260 Infinity Fraction Collectors User Manual upper 67 tick marks 77 time b
68. finity Fraction Collectors User Manual Safety Information A Lithium Batteries Information Danger of explosion 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 Fraction Collectors User Manual 151 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
69. gt 2COClg 2HCI This reaction in which stainless 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 Fraction Collectors 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 Fraction Collectors User Manual 155 A Safety Information Agilent Technologies on Internet 156 Agilent 1260 Infinity Fraction Collectors User Manual Index A above location 46 accessory kit 140 accessory kit contents 16 adjusting the tray height 24 Agilent on internet 155 air circulation 11 analytical needle assembly 114 applicati
70. he bench in all horizontal position 8 Ensure the power switch at the front of the Fraction Collector is OFF Connect the power cable to the power connector at the rear of the Fraction Collector 10 Connect the CAN cable to the other Agilent 1260 Infinity LC modules 11 If an Agilent ChemStation is the controller it must be connect 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 fraction collector The test tube trays can be adjusted in height depending on the height of the used test tubes To adjust the height of the trays press against the snappers at Agilent 1260 Infinity Fraction Collectors User Manual 23 1 Installing the Fraction Collector the guides in the 4 corners of a tray and move the top plate of the tray up or down see Figure 6 on page 24 The preparative scale fraction collector G1364B and the analytical scale fraction collector G1364C are shipped with two trays 40x2 ml half tray and 15 x 6ml half tray Figure 6 Adjusting the Height of the Full Test Tube Trays 14 Turn ON power by pushing the button at the lower left hand side of the Fraction Collector 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 Fraction Collector will start the hardware initiali
71. hese assemblies into the appropriate maintenance position In the ChemStation the fraction collector maintenance positions can be selected from the Maintenance menu in the Diagnosis view see Figure 38 In the Agilent Instant Pilot G4208A the functions can be selected in the Test screens of the fraction collector Fraction collector Maintenance Positions x rChange Parts End r Move Arm Home Park Arm for Transport lt Figure 38 Fraction Collector Maintenance Positions dialog box Change Parts If you click Start the transport unit will move upwards the needle carrier assembly will move to the front center and then turn OFF the theta motor to allow free rotation of the theta arm This position enables easy access to the transport unit to change Agilent 1260 Infinity Fraction Collectors User Manual 95 3 Troubleshooting and Test Functions e The needle assembly e The needle carrier assembly The valve to needle tubing e The diverter valve After the maintenance or repair task has been finished click End to move the transport assembly to the Home position 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 Fraction Collector 96 Agilent 1260 Infinity Fraction Collecto
72. ilent 1260 Infinity DAD MWD G1315D G1365D Peakwidth min Response time sec Signal delay sec lt 0 01 gt 0 01 gt 0 03 gt 0 05 gt 0 10 gt 0 20 gt 0 40 gt 0 85 0 1 0 2 0 5 1 0 2 0 4 0 8 0 16 0 0 05 0 15 0 5 1 25 2 75 5 9 11 9 23 9 Agilent 1260 Infinity Fraction Collectors User Manual 51 2 52 Configuration and Operation of the Fraction Collector Table 10 Signal Delay Times for the Agilent 1260 Infinity DAD MWD 1315C G1365C Peakwidth min Response time sec Signal delay sec lt 0 0025 gt 0 0025 gt 0 005 gt 0 01 gt 0 03 gt 0 05 gt 0 1 gt 0 2 gt 0 4 gt 0 85 0 02 0 05 0 1 0 2 0 5 1 0 2 0 4 0 8 0 16 0 0 0375 0 0625 0 144 0 294 0 619 1 27 2 57 5 17 10 4 20 8 Table 11 Signal Delay Times for the Agilent 1260 Infinity VWD G1314B Peakwidth min Response time sec Signal delay sec lt 0 005 gt 0 005 gt 0 01 gt 0 025 0 05 0 1 0 2 0 4 lt 0 12 0 12 0 25 0 5 1 2 4 8 0 07 0 14 0 29 1 3 2 8 6 0 12 3 Agilent 1260 Infinity Fraction Collectors User Manual Configuration and Operation of the Fraction Collector 2 Table 12 Signal Delay Times for the Agilent 1260 Infinity VWD G1314C Peakwidth min Response time sec Signal delay sec lt 0 00125 gt 0 00125 gt 0 0025 gt 0 005 gt 0 01 gt 0 025 gt 0 05 gt 0 1 gt 0 2 gt 0 4 lt 0 02 0 02 0 05 0 12 0 25
73. ing large tt Reorder gives 10 clamps for waste tubing small 140 Agilent 1260 Infinity Fraction Collectors User Manual Agilent 1260 Infinity Fraction Collectors User Manual 6 Specifications Performance Specifications for the Fraction Collectors 142 of Agilent Technologies 141 6 Specifications Performance Specifications for the Fraction Collectors 142 Table 35 performance Specifications Agilent 1260 Infinity Preparative scale Fraction Collector G1364B Type Specification Trigger modes Operating modes Fraction capacities and trays Test tube plate sizes Maximum tube volume Time slices Peak threshold up downslope Timetable combination of time intervals and peak and Manual trigger supported only with Agilent Instant Pilot G4208A Agilent 1260 Infinity DAD MWD detectors G1315C D and G4212B G1365C D the Agilent 1260 Infinity Fluorescence Detector G1321A and the Agilent G1946C D G1956A B LC MSD are fully supported other detectors can be used but are not supported for fraction collection Discrete fractions default mode for all vessels The flow is diverted to waste while moving from one vessel position to the next vessel position Continuous flow optional available only when using well plates It is possible to move from one well plate position to the next one without diverting the flow into the well plate to waste 4 x well plates full tray MTP for use with deep
74. inity High Performance Autosampler and fraction collection will start at Plate 1 Position A1 of the fraction collector For the next Sample 2 which is injected from a different location in the high performance autosampler the fraction collection will start again at Plate 1 Position A1 of the fraction collector Line Location Sample Name Dilution Datafile Inj Yolume Frac Start Prat Sampe ss CE iam P1A02 Sample2 CO CAO Sample3 J CT o Cd PAO Figure 33 Pooling Sequence If you specify multiple injections for pooling the overfill protection is not activated It is the user s responsibility to make sure that all fraction collection locations are large enough to completely collect all pooled fractions If a fraction collection location is overfilled an error message occurs and the pump is shut OFF 78 Agilent 1260 Infinity Fraction Collectors User Manual Configuration and Operation of the Fraction Collector 2 Sample Recovery The Agilent 1260 Infinity Fraction Collectors offer different possibilities for sample recovery e The preferred recovery strategy is to install multiple fraction collectors in your LC systems and use the last of those fraction collectors for recovery This recovery fraction collector can be selected in the Configuration dialog box as displayed in Figure 34 The fourth fraction collector in your systems will always be used for sample recovery Also read Configuring Multiple Fraction Collect
75. inless steel flow cell G1315 60016 For other pre parative flow cells some parameters e g flow rate injection volume threshold slope should be adjusted Expected Result Run a single injection under the conditions as described in Table 15 You should observe a chromatogram with four peaks as displayed in Figure 36 on page 86 Four fractions should have been collected in the fraction collector vials The retention times of the peaks could be different depending on the system configuration e g flow cell type capillary volumes type and age of column Agilent 1260 Infinity Fraction Collectors User Manual Configuration and Operation of the Fraction Collector 2 Application Notes More information about the Agilent 1260 Infinity Fraction Collectors and Purification Systems are available from a series of application notes Printed versions can be ordered from Agilent or pdf files can be downloaded from the Agilent Website http www chem agilent com Table 16 Selected Agilent Technologies Application Notes Title Publication Number An optimized system for analytical and preparative work 5988 9649EN Recovery collection with the Agilent LC purification system 5988 9650EN Solutions for preparative HPLC Application Compendium 5988 9646EN Innovative fraction collection with the Agilent LC purification platform 5988 9250EN Automated fraction re analysis does it really make sense 5988 8653EN Injection of high concentration sa
76. ion Data File p 1 3 2006 2 31 28 Vial 1 CHECKOUTAS M o jo i E CHECKOUTO DAD1 A Sig 254 16 Ref 360 100 SCREENS CHECKOUTOO0001 D mAU 6004 5004 400 300 2004 coe REE amp 3 CE SPPPPPPPEEPPPPPPEERRPEER eer sai N E indi eda ae EE eo ane ene ee EE EAE E eras r r r T r o 1 4 6 min D File Information pii File Path C CHEM32 1 DATA SCREENS al Fraction Fraction Location _ Start Time min End Time min _Volume pl Reason Date 03Jan 06 15 31 28 1 1 P1 A 01 1 75 t 187 71 DAD1 Sample I 2 1 P1 4 02 214 227 148 33 DAD1 Sample Into 3 1 P1 4 03 3 25 3 42 198 54 DAD1 Barcode 4 1P14 04 4 87 512 268 33 DAD1 Operator Method CHECKOUTAS M Analysis Time 7 133 min Sampling Rate 0 0033 min 0 198 sec 2141 datapoints al Ready 0 0033 min 0 198 sec 2141 datapoints Figure 36 Chromatogram of the check out injection with UV based trigger Agilent 1260 Infinity Fraction Collectors User Manual Configuration and Operation of the Fraction Collector 2 Check out Procedure for UV triggered Fraction Collection PS Prerequisites V The Agilent 1260 Infinity LC System has been flushed thoroughly with the liquid phase as described in Table 14 and the lamp s of the UV detector have been switched on for at least 15 minutes to provide a stable baseline V The Agilent 1260 Infinity Fraction Collector has been installed and configured The delay volume has been calibrated as described in Performi
77. le carrier assembly in case it slipped out of this guide see 2 left arrows on the figure to the left It is absolutely vital that the tubing is installed as described to maximize the lifetime of the tubing Re install the tray s in the tray base Start the instrument Close the front cover Agilent 1260 Infinity Fraction Collectors User Manual 113 4 Repairing the Fraction Collector Exchanging the Analytical Needle Assembly Frequency When the needle is visibly damaged When the needle is blocked or contaminated Tools required Wrench open end 4mm PN 8710 1534 supplied in accessory kit Wrench open end 1 4 5 16 inch PN 8710 0510 supplied in accessory kit Parts required Analytical needle assembly G1367 87200 WARNING To avoid personal injury keep fingers away from the needle area during Fraction Collector operation CAUTION Regularly inspect the inlet waste tubing assembly and the valve to needle tubing and exchange them if they are worn out or show visible signs of damage Before beginning this procedure 1 Use the 4 mm wrench and the 5 16 wrench for counter holding to unscrew the valve to needle tubing from the needle assembly Viewed from the bottom ZB af SS Position the transport unit of the Fraction Collector in the Home Position see Maintenance Functions on page 95 Remove all installed trays from the tray base Position the transport unit of the F
78. lector CAUTION If you need to ship the Fraction Collector at a later date always use the shipping protection foam parts see Transporting the Fraction Collector on page 41 Damaged Packaging CAUTION Upon receipt of your Fraction Collector 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 Fraction Collector 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 Fraction Collector please do not attempt to install the Fraction Collector Delivery Checklist 14 Ensure all parts and materials have been delivered with the Fraction Collector 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 Three models of Agilent 1260 Infinity fraction collectors are available G1364B Fraction Collector preparative scale designed for flow rates up to 100 ml min and for the use with vials deep well plates and test tubes up to 100 mm height G1364C Fraction Collector analytical scale designed for flow rates below 10 ml min can be modified for higher flow rates and for the use with
79. lied 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 Fraction Collector operation Do not attempt to insert or remove a vial or a plate when the needle is positioned 22 Install the LAN interface board in the Fraction Collector if 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 a A Ww 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 5 on page 23 Route the corrugated waste tubing into a waste container Agilent 1260 Infinity Fraction Collectors User Manual Installing the Fraction Collector 1 Figure 5 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 Fraction Collector in the stack or on t
80. llector Agilent 1260 Infinity Fraction Collectors User Manual Agilent 1260 Infinity Fraction Collectors User Manual 2 Configuration and Operation of the Fraction Collector Configuration of the Fraction Collector 44 Delay Volumes and Delay Calibration 50 Setting up a Fraction Collector Method 65 Starting Your Run with Fraction Collection 72 Viewing Your Results 76 Special Applications 78 Limitations and How to Avoid Problems 83 Check out Procedures 84 Application Notes 89 ee Agilent Technologies 43 2 Configuration and Operation of the Fraction Collector Configuration of the Fraction Collector Configuration of the Fraction Collector in the ChemStation CAUTION Before using the preparative scale fraction collector G1364B the delay calibration adapter G1364 87301 has to be removed from the needle carrier assembly 44 In order 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 resulting Fraction Collector Configuration dialog box Figure 15 on page 45 the configuration of the Trays the Fraction Delay Volumes the Collection Order the Needle Movement and the Well Plates can be specified Trays In the online mode of the Agilent ChemStation the type of the installed tray is recognized automatically In the off line mode an appropria
81. lue is set to 0 5 minutes If broad peaks are expected this value should be increased without exceeding the run time Threshold Max Peak Duration Figure 25 Maximum Fraction Duration Peak Detectors In the Peak Detectors section a list of all Peak Detectors that are connected to the system is displayed Agilent 1260 Infinity Diode Array Detectors multi wavelength detectors variable wavelength detectors and fluorescence Agilent 1260 Infinity Fraction Collectors User Manual Configuration and Operation of the Fraction Collector 2 detectors are recognized automatically Other Detectors e g Agilent 6000 Series mass selective detectors or HP1050 Detectors are connected through the Universal Interface Box UIB The Peak detector table contains seven columns Working Mode For each peak detector Threshold only Threshold Slope or 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 slope of the detector signal 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 val
82. ly Preparative Scale partly displayed 134 Agilent 1260 Infinity Fraction Collectors User Manual Parts and Materials 5 Transport Unit Assembly Analytical Scale Table 29 Transport Unit Assembly Analytical Scale Item Description Part Number 1 Transport unit assembly analytical scale G1364 60019 includes items 2 and 4 2a b Tubing diverter valve to needle 0 25 mm ID G1364 68712 order full kit 3 Diverter valve assembly G1364 61901 4 Needle carrier assembly analytical scale 61364 60022 5 Needle assembly analytical scale 50mm needle G1367 87200 Needle assembly semi preparative scale 20mm needle 61364 87202 6 Wellplate adapter G1364 60021 Figure 43 Transport Unit Assembly Analytical Scale partly displayed Agilent 1260 Infinity Fraction Collectors User Manual 135 5 Parts and Materials Needle Assemblies Table 30 Needle Assemblies Item Description Part Number 1 Needle assembly preparative scale G1364 87201 2 Needle assembly semi preparative scale 20 mm needle G1364 87202 For use in the analytical scale fraction collector at flow rates gt 10 ml min or with high test tubes 48 mm 75mm 3 Needle assembly analytical scale 50 mm needle G1367 87200 Delay Calibration Adapter not shown G1364 87301 Figure 44 Needle Assemblies 136 Agilent 1260 Infinity Fraction Collectors User Manual Parts and Materials Diverter Valve Assembly Table 31 Diverter Val
83. mples with the Agilent LC purification 5988 8654EN system Sophisticated peak based fraction collection working with up and 5988 7895EN down slope Strategies for purification of compounds from non baseline separated 5988 7460EN peaks Method scale up from analytical to preparative scale with the Agilent LC 5988 6979EN purification system PS Peak based fraction collection with the Agilent LC purification system 5988 5747EN AS Influence of delay volume on recovery Agilent 1260 Infinity Fraction Collectors User Manual 89 2 Configuration and Operation of the Fraction Collector Application Notes 90 Agilent 1260 Infinity Fraction Collectors User Manual Agilent 1260 Infinity Fraction Collectors User Manual 3 Troubleshooting and Test Functions Status Indicators 93 Maintenance Functions 95 Maintenance Functions 95 Transport Unit Self Alignment 97 Step Commands 99 ee Agilent Technologies 91 3 Troubleshooting and Test Functions Status Indicators The Fraction Collector is provided with two status indicators which indicate the operational state prerun not ready run and error states of the instrument The status indicators provide a quick visual check of the operation of the Fraction Collector see Status Indicators on page 93 Error Messages In the event of an electronic mechanical or hydraulic failure the instrument generates an error message in the user interface For a detailed description of th
84. ng a Delay Calibration with an UV Detector on page 55 V The 15 vial tray G1313 44513 with at least four 6ml vials is installed 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 15 vial tray Then click on Yes in displayed dialog box Table 15 Method Parameters for UV triggered Fraction Collection Check out PS Agilent 1260 Infinity modules preparative pump plus gradient extension optional preparative autosampler dual loop autosampler e diode array detector or multi wavelength detector with preparative flow cell e fraction collector preparative scale Column ZORBAX SB C18 50 mm x 9 46mm 846975 202 or equivalent Sample Check out sample 01080 68704 Flow 10 ml min Stoptime 4 minutes Solvent A 35 HPLC grade water Solvent B 65 HPLC grade acetonitrile Wavelength DAD MWD Signal A 254 16nm Reference 360 80nm Peakwidth Responsetime gt 0 1min 2s Injection volume 50 pl Column temperature ambient Fraction trigger mode Peak based Agilent 1260 Infinity Fraction Collectors User Manual 87 2 88 Configuration and Operation of the Fraction Collector Table 15 Method Parameters for UV triggered Fraction Collection Check out PS Max peak duration 0 5 min DAD MWD working mode Threshold slope Upslope 5 mAU s Downslope 10 mAU s Threshold Upper threshold 10 mAU 3000 mAU The checkout method is tailored for the 3mm sta
85. nt cover and route the condensation drain tube to a waste container 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 fraction collector thermostat requires 25 cm 10 inch space on each for sufficient air circulation 26 Install the LAN interface board in the Fraction Collector if 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 nun OO ol FF Q Install the corrugated waste tube in the plastic port at the front bottom center of the Fraction Collector and route down into a waste container Slide the waste tubing coming from the internal tray Gif present through Agilent 1260 Infinity Fraction Collectors User Manual Installing the Fraction Collector 1 the plastic port and the corrugated waste tube see Figure 5 on page 23 Route the corrugated waste tubing into a waste container 8 Re install the left side door take care of the magnet at the back
86. nt to store the new calculated delay volume for the UV detector in the Fraction Collector Configuration see also Configuration of the Fraction Collector in the ChemStation on page 44 25 Press Print to get a hard copy of the report 26 Stop the splitter 27 Return to the Method and Run Control View and load the method you will be using to run your samples Select Fraction Collection from the More MSD sub menu under the Instrument menu Press Parameters and enter the final calculated delay time that appears in the report in the Collector Delay field first converting it to minutes In this case you would enter 11 991 60 0 20 minutes see Figure 23 62 Agilent 1260 Infinity Fraction Collectors User Manual Configuration and Operation of the Fraction Collector 2 Delay Volumes and Delay Calibration Fraction Collection Parameters Figure 23 Fraction Collection Parameters dialog box with delay volume entry 28 Save the method NOTE If you anticipate running samples at a variety of flow rates it is best to calibrate at the highest likely flow rate first and install the appropriate length tubing This way there will be sufficient delay for the lower flow rates and new lengths will not need to be cut Agilent 1260 Infinity Fraction Collectors User Manual 63 2 Configuration and Operation of the Fraction Collector CAUTION Before using the preparative scale fraction collector G1364B the delay calibration
87. on Collectors User Manual 97 3 Troubleshooting and Test Functions How to perform a Transport Unit Self Alignment Steps Comments 3 Ensure that the wellplate adapter is correctly On a G1364B preparative scale fraction assembled collector install the delay calibration adapter G1364 87301 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 8 bit configuration switch in the setting Fraction Collectors Service Manual If the Transport Unit Self Alignment is started with well plates on the tray the alignment procedure is aborted without any error message 98 Agilent 1260 Infinity Fraction Collectors User Manual Troubleshooting and Test Functions 3 Step Commands Some movements of the fraction collection sequence can be done under Manual control This is useful during troubleshooting where close observation of each of the fraction collection step 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 Fraction Collector components to predefined positions enabling the specific step to be done In the ChemStation the step commands can be selected f
88. on notes 89 arm length calibration 99 battery safety information 151 bench space 11 C cable connecting APG remote 19 21 connecting CAN 19 21 connecting GPIB 19 21 connecting LAN 19 21 connecting the ChemStation 19 21 connecting the power 19 21 cable connections 25 capacities 142 145 capillaries 31 check out procedure 84 ChemStation configuration 44 cleaning the fraction collector 102 closing mats 132 collection mode 46 collection order 46 condensation 12 configuration 44 collection order 46 fraction delay volumes 46 needle movement 46 trays 44 well plates 47 configuring well plate types 37 contact control 46 cooling 143 146 corrugated waste tubing 23 D damaged packaging 14 data analysis 76 delay calibration sensor 143 145 delay volume 46 configuration 46 delay volumes 143 145 delivery checklist 14 detectors 66 diverter valve 143 145 diverter valve assembly 137 E electrical connections thermostat 30 environment 10 12 error messages 92 95 F failure 92 fill levels reset 72 fill volume maximum 69 Agilent 1260 Infinity Fraction Collectors User Manual flow connections 31 fraction capacities and trays 142 fraction delay volumes 46 fraction list 76 fraction preview 70 fraction vessel capacities and trays 145 funnel 121 fuses 10 G GLP features 143 146 half tray combinations 35 height 24 home posi
89. ors on page 48 Fraction Collector 2 Configuration Instrument 1 x Trays Tray 40 Funnels M IV Use for Sample Recovery Tray B 40 Funnels M Tube Volume ml 0 00 Tray C 40 Funnels M Tube Height mm jo Figure 34 Sample Recovery Fraction Collector For the following tray configurations fixed recovery location will be assigned from the ChemStation In order to disable the recovery the tray configuration has to be changed 1 Standard tray for two well plates 10 2ml x vials G2258 60011 and half tray for 40 x 2 ml vials G1313 44512 In this configuration the forty 2 ml vials on the half will automatically be used for recovery 2 Standard tray for two well plates 10 x 2ml vials G2258 60011 and half tray for 15x 6 ml vials G1313 44513 In this configuration the fifteen 6 ml vials on the half will automatically be used for recovery 3 Standard tray for two well plates 10 x 2ml vials G2258 60011 and half tray with 40 funnels G1364 84532 In this configuration the funnels on the half will automatically be used for recovery e The Standard tray for two well plates 10 funnels G1364 84522 allows two modes of operation Agilent 1260 Infinity Fraction Collectors User Manual 79 2 Configuration and Operation of the Fraction Collector CAUTION 1 If well plates are configured in the fraction collector configuration the checkbox Plate 1 or Plate 2 are selected the 10 funnel locations will
90. r 001 0101 D 3 Ls T i it 02 0 4 06 DAD1 A Sig 600 10 Ref off 001 0101 D 4 Figure 20 Delay Calibration Results Panel The Calibration Results panel contains a results table and the acquired chromatograms from the detector s and the fraction collector delay sensor The results table contains e the name and serial number of the detector e the current delay volume stored in the fraction collector configuration e the calculated delay volume from the calibration To print the Calibration Results panel click Print 11 If you want to store the calculated delay volume in the fraction collector configuration click Calibrate A confirmation dialog box is displayed when the delay volume has been stored to close the confirmation click OK If you do not want to store the calculated delay volume in the fraction collector configuration click the Close button A confirmation dialog box is displayed click OK to close the Confirmation or Cancel to return to the Calibration Results panel Agilent 1260 Infinity Fraction Collectors User Manual 57 2 Configuration and Operation of the Fraction Collector CAUTION The Delay Calibration can only be performed for fraction collector 1 of your configured system The calculated delay volume is used for all fraction collectors therefore the volume tubing between the selection valve and each installed fraction collector must be identical
91. r Manual 4 Repairing the Fraction Collector Cleaning the Fraction Collector 102 Overview of Main Repair Procedures 103 Simple Repairs 104 s f Agilent Technologies 101 4 Repairing the Fraction Collector Introduction into Repairing the Fraction Collector Simple Repairs The Fraction Collector is designed for easy repair The most frequent repairs such as changing a needle assembly or tubings 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 on page 104 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 To avoid personal injury keep fingers away from the needle area during Fraction Collector operation Do not bend the safety flap away from its position or attempt to insert or remove a vial from the gripper when the gripper is positioned below the needle Regularly inspect the inlet waste tubing assembly and the valve to needle tubing and exchange them if they are worn out or show visible signs of damage Cleaning the Fraction Collector The Fraction Collector covers should be kept clean Cleaning should be done with a soft cloth slightly dampened with water o
92. r a solution of water and a mild detergent Do not use an excessively damp cloth from which liquid could drip into the Fraction Collector Do not let liquid drip into the Fraction Collector It could cause a shock hazard or damage to the Fraction Collector 102 Agilent 1260 Infinity Fraction Collectors User Manual Overview of Main Repair Procedures BCD Board optional Main board T K q Needle carrier Q Needle Wellplate Adapter Plug channel with air channel adapter underneath Figure 40 MA ny ee EE CA A WEA Repairing the Fraction Collector 4 Transport unit Diverter Valve Ribbon cable Inlet waste tubings Internal tray with rinse port and flow delay sensor Leak plane with leak sensor ray compartment divider Main Assemblies Example displays the analytical scale module G1364C The preparative scale fraction collector G1364B contains a different needle Agilent 1260 Infinity Fraction Collectors User Manual 103 4 Repairing the Fraction Collector Simple Repairs The procedures described in this section can be done with the Fraction Collector in place in the stack These procedures can be done on a more frequent basis Table 18 Simple Repair Procedures Procedure Typical Frequency Notes Replacing the Inlet waste tubings Replacing the valve to needle tubing Exchanging the preparative needle assembly Exchanging the analytical needle
93. r each of those fraction collectors For the recovery fraction collector the delay volume parameter will be ignored The Delay Calibration can only be performed for fraction collector 1 of your configured system The calculated delay volume is used for all fraction collectors therefore the volume tubing between the selection valve and each installed fraction collector must be identical Otherwise fraction collector 2 and fraction collector 3 have to be temporarily configured as fraction collector 1 during the delay calibration Agilent 1260 Infinity Fraction Collectors User Manual Configuration and Operation of the Fraction Collector 2 The fraction collector inlet tubings have to be connected to port 1 3 of the selection valve in the same order as configured in the ChemStation The recovery fraction collector is always connected to the waste tubing of the main fraction collectors The waste tubing from multiple fraction collectors and the inlet tubing to the recovery collector have to be connected through a T Piece e Fraction Collector 4 will always be used as a recovery fraction collector Agilent 1260 Infinity Fraction Collectors User Manual 49 2 Configuration and Operation of the Fraction Collector Delay Volumes and Delay Calibration 50 Delay Times and Volumes Figure 16 shows a schematic drawing of the flow path between the detector and the fraction collector with the two delay volumes Vp and Vp For peak bas
94. r supply indicator is integrated into the main power switch When the indicator is illuminated green the power is ON Agilent 1260 Infinity Fraction Collectors User Manual 93 3 94 Troubleshooting and Test Functions Instrument Status Indicator The instrument status indicator indicates one of four possible instrument conditions e 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 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 Agilent 1260 Infinity Fraction Collectors User Manual Troubleshooting and Test Functions 3 Maintenance Functions Some maintenance procedures require the needle arm and needle carrier to be moved to specific positions to enable easy access to components The maintenance functions move t
95. raction Collector in the Change Parts Position see Maintenance Functions on page 95 Turn OFF the instrument It might be more convenient to remove the needle from its carrier before unscrewing the needle tubing 114 Agilent 1260 Infinity Fraction Collectors User Manual Repairing the Fraction Collector 4 2 Remove the wellplate adapter by turning it clockwise 1 Hold the needle assembly between your thumb and forefinger slide out the assembly towards the rear of the needle carrier assembly 2 N ASS x VS Z TS DN yal Co Ze sR 3 Insert the new needle assembly into the holder of the needle carrier assembly Make sure to push it all the way to the front Reassemble the wellplate adapter 4 Use the 4 mm wrench and the 5 16 wrench for counter holding to connect the valve to needle tubing to the needle Reassembly the wellplate adapter Viewed from the bottom Cig pa VAL LA On completion of this procedure IMPORTANT After fixing the needle in the needle carrier clip the tubing into the guide on the bottom of the needle carrier assembly in case it slipped out of this guide see 2 left arrows on the figure to the left It is absolutely vital that the tubing is installed as described to maximize the lifetime of the tubing Re install the tray s in the tray base Start the instrument Close the fron
96. ray Compartment Divider 12 Install the tray you have ordered for your fraction collector The test tube trays can be adjusted in height depending on the height of the used test tubes To adjust the height of the trays press against the snappers at the Agilent 1260 Infinity Fraction Collectors User Manual Installing the Fraction Collector 1 guides in the 4 corners of a tray and move the top plate of the tray up or down see Figure 6 on page 24 13 Ensure the power switch on the front of the Fraction Collector is OFF and the power cables are disconnected 14 Connect the cable between the Fraction Collector and the thermostat see Figure 10 on page 30 Do not disconnect or reconnect the Fraction Collector to thermostat cable when the power cords are connected to either of the two modules This will damage the electronics of the modules 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 connect to the LAN interface should be installed to the detector 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 Fraction Collector 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 Fraction Collector will
97. rd Do not install substitute parts or make any unauthorized modification to the instrument Capacitors inside the instrument may still be charged 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 Fraction Collectors User Manual 149 A Safety Information Safety Symbols Table 37 shows safety symbols used on the instrument and in the manuals Table 37 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 lamp used in this product Always turn the xenon flash lamp off before removing it Q Eye damage may result from directly viewing the light produced by the Xenon flash 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 150 Agilent 1260 In
98. rom the Test Selection Box see Figure 39 in the Diagnosis display In the Agilent Instant Pilot G4208A the step commands can be accessed from the pull down menu in the Fraction Collector Test Automatic Fraction Collector Steps Valve to Waste Valve to Vessel Needle Up Move to Location Vial 1 Move to Rinse Needle into Vessel Figure 39 Fraction Collector Step Commands Agilent 1260 Infinity Fraction Collectors User Manual 99 3 100 Troubleshooting and Test Functions Table 17 Step Commands Step Action Comments Needle Up Needle into vessel Needle to rinse flush port Switch valve to needle Switch valve to waste Lifts the needle arm to the upper position Lowers the needle into the specified vessel Moves the needle to the rinse flush port Switches the diverter valve to the needle Switches the diverter valve from needle to waste Command also switches the diverter valve to waste if it is not already in that position only analytical scale G1364C only analytical scale G1364C Troubleshooting If the Fraction Collector is unable to perform a specific step due to a hardware failure an error message is generated You can use the step commands to perform a fraction collection sequence and observe how the Fraction Collector responds to each command Agilent 1260 Infinity Fraction Collectors User Manual Agilent 1260 Infinity Fraction Collectors Use
99. rs User Manual Troubleshooting and Test Functions 3 Transport Unit Self Alignment The transport unit alignment with the transport unit and the well plate tray is required to compensate for larger deviations in positioning the needle carrier assembly The transport unit self alignment is required after disassembling the system or when you exchange the transport unit the needle 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 one of the 4 well plate trays G1364 84501 but well plates MUST NOT be installed 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 needle capillary carrier assembly e The MTP main board How to perform a Transport Unit Self Alignment Steps Comments 1 If the transport unit has been exchanged or ifit For details see Forced Cold Start in the is very badly misaligned set the 8 bit Fraction Collectors Service Manual configuration switch to the Forced Cold Start Configuration 2 Install the 4 well plate tray G1364 84501 IMPORTANT Remove all plates Agilent 1260 Infinity Fracti
100. st way by row e shortest way by column Shortest way by row column is recommend if Continuous flow is selected as Collection mode Collection Mode The Collection Mode defines the triggering of the diverter valve For Discrete fractions the diverter valve switches to waste while the needle moves to the next test tube vial or well For Continuous flow the diverter valve is not switching during needle movements except for long movements in the row by row or column by column mode In addition the needle cannot move Into location during continuous flow operation Continuous flow is only available for well plates Needle Movement Into location In the Into Location mode the needle tip moves into the well to the specified Depth in mm This option is only available for the analytical scale fraction collector in the Discrete fractions mode Above location In the Above Location mode the needle tip stays at the specified Distance in mm above the well during fraction collection Contact Control In this mode the needle tip moves down to the well bottom until is reaches the specified Distance in mm between needle tip and the vial well bottom This ensures that the forming droplet is in contact to the Agilent 1260 Infinity Fraction Collectors User Manual Configuration and Operation of the Fraction Collector 2 vial well bottom During the continuing filling process the needle tip moves upwards while staying in contact with the surface of
101. t 0 05 min collection Peak based Set threshold and slope values such that length of fractions is at least 0 05 fraction min collection Unresolved peaks can be separated using appropriate threshold and slope values If two unresolved peaks are to be collected as one fraction collect based on threshold only e Ifthe baseline of the chromatogram is below or above 0 mAU this offset is not accounted for when triggering peaks using a threshold value The threshold value is always added to 0 mAU 82 Agilent 1260 Infinity Fraction Collectors User Manual Configuration and Operation of the Fraction Collector 2 Limitations and How to Avoid Problems Rinse Fraction If Rinse Fraction Collection Needle is set to Between fraction collection at Collection Needle least 0 3 min are required to perform this task When doing time based fraction collection rinsing the needle is only possible between two time table entries which must have a gap of at least 0 3 min For peak based fraction collection a time gap of also at least 0 3 min is required If a new peak is detected during the rinse process it is aborted and the needle moves back to the next free fraction position Depending on flow rate and delay volume Vp the beginning of this peak may be lost If you have recovery positions in your fraction collector or if you are using one fraction collector for sample recovery in a multiple fraction collector configuration the rinse function betwe
102. t cover Agilent 1260 Infinity Fraction Collectors User Manual 115 4 Repairing the Fraction Collector Exchanging the Diverter Valve Frequency When leaky or defective Tools required Hex key 2 0 mm PN 8710 2438 supplied in accessory kit Parts required Diverter Valve PN G1364 61901 Before beginning this procedure 1 Locate the diverter valve with the finger tight fittings of the inlet waste tubing assembly the figure shows the open Position the transport unit of the Fraction Fraction Collector seen from the front Collector in the Home Position see Maintenance Functions on page 95 Remove all installed trays from the tray base Position the transport unit of the Fraction Collector in the Change Parts Position see Maintenance Functions on page 95 Turn OFF the instrument 2 Unscrew the 2 finger tight fittings of the inlet waste 3 Unscrew the finger tight fitting of the valve to needle tubing assembly at the diverter valve tubing assembly at the diverter valve TE i maa Dal 7 Le lo 116 Agilent 1260 Infinity Fraction Collectors User Manual Repairing the Fraction Collector 4 4 Disconnect the power cable of the diverter valve from the 5 Using the 2 mm hex key loosen the holding screw of the connector on the front of the z arm diverter valve and remove
103. tal duration of time table time of last entry OFF fraction collector delay time Vp v 0 1min for time based fraction collection End of last peak tg fraction collector delay time Vp v 0 1min_ for peak based fraction collection Agilent 1260 Infinity Fraction Collectors User Manual Configuration and Operation of the Fraction Collector 2 Performing a Delay Calibration with an UV Detector 1 Place a vial containing the Delay Sensor Calibrant G1946 85020 in position 1 of the Autosampler 2 Remove the installed column and connect the capillaries with a zero dead volume connector 3 Install the 40 x 2ml vial tray analytical scale or the 15 x 6ml vial tray preparative scale in the fraction collector 4 Connect a bottle of water to Channel A 5 Switch to Diagnosis View if necessary 6 Open the AFC Delay Volume Calibration status window see Figure 18 from the Fraction Collector sub menu of the Maintenance menu Maintenance gt Fraction Collector gt Delay Volume Calibration AFC Delay Yolume Calibration Status Window turn on all pumps Figure 18 Delay Volume Calibration status window 7 The Info field gives the instruction to turn ON the pumps see Figure 18 Click Continue Agilent 1260 Infinity Fraction Collectors User Manual 55 2 56 Configuration and Operation of the Fraction Collector AFC Delay Volume Calibration Status Window Calibration method DGCALAS M
104. te tray type can be chosen from a dropdown list For a selection of supported trays see Supported Trays for Fraction Collectors on page 126 Tube volume ml Volume of the test tube should be specified 10 less than the maximum fill volume to avoid overfill Tube height mm The exact test tube height has to be specified For preparative scale fraction collector the minimum height is 48 mm and the maximum height is 100 mm For the analytical scale fraction collector with 50 mm needle standard the maximum height is 48 mm By changing to the short 20 mm needle the maximum tube height can be in creased to 75 mm For the part numbers of the different needle assemblies see Needle Assemblies on page 136 In the online ChemStation the installed tray is recognized and the default settings for tube volume and tube height are loaded and displayed Agilent 1260 Infinity Fraction Collectors User Manual Configuration and Operation of the Fraction Collector Fraction Collector Configuration LC113 x Trays Tray A 40 Tubes Tray B Tray C Tube Volume ml 45 00 Tube Height mm 98 Fraction Delay Volumes Detector VYolume ul a Column by Ill C Shortest path column by row Collection Order C Row by row O Shortest path Collection Mode Discrete fractions Continuous flow Well Plates Needle Movement into location Depth 5 0 mm above location Distance 2 0 mm
105. th low internal volume 15 ul switching time lt 100 ms maximum operating pressure 6 bar Optional with additional G1330B performance depending on ambient conditions and the volume of collected fractions 3 fraction collectors in parallel plus one recovery fraction collector connected via 12 Position 13 Port Selector valve PN G1160A Early maintenance feedback EMF electronic records of maintenance and errors 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 Leak detection and safe leak handling error detection and display exhaust fan for fume extraction of hazardous vapors Vials can be used as recommended by Agilent Technologies see List of Recommended Vials and Caps on page 129 and List of Recommended Plates and Closing Mats on page 132 but must be uncapped Only the 96 deep well plates can be used without closing mats see List of Recom mended Plates and Closing Mats on page 132 Only one type of well plates can be used at a time in one tray Agilent 1260 Infinity Fraction Collectors User Manual 143 6 144 Specifications Table 36 Performance Specifications Agilent 1260 Infinity Analytical scale Fraction Collector G1364C Type Specification Trigger modes Opera
106. the Fraction Preview screen By pushing the desired buttons on the right hand side of Fraction Preview screen the chromatogram can be Agilent 1260 Infinity Fraction Collectors User Manual Configuration and Operation of the Fraction Collector 2 Setting up a Fraction Collector Method 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 Fraction Collectors User Manual 71 2 Configuration and Operation of the Fraction Collector Starting Your Run with Fraction Collection CAUTION Resetting the Current Fill Levels To protect the collected fractions against contaminations and to avoid overfilling of single positions the Agilent 1260 Infinity Fraction Collector stores current fill levels It will block used positions if no fraction start location has been specified If a new tray of the same type is inserted in the fraction collector all fill levels can be reset not just the well plate vial or test tube After re installation of the tray a pop up window as displayed in Figure 27 will be displayed Click on Yes to reset all fill levels from the previous run p Tray 1 was replaced by a tray of the same type 7 Do you want to reset the fill volume Yes No Figure 27 Reset fill volumes The window Figure 27 to reset current fill levels is not displayed if
107. the collected liquid This option is recommend for low flow rates and small fraction volumes to avoid air bubbles and accomplish an accurate fraction volume It is only available for the analytical scale fraction collector Well Plates In the Well Plates section the type of well plates used in a well plate tray can be configured The well plate type can be chosen from the Plate Type dropdown list More detailed information about all pre configured well plates can be found in the Instrument menu Only one type of well plate can be used on the well plate tray The checkbox Treat as 4 x 96 is available only for 384 well plates and allows to split the 384 well plate virtually into 4 separate 96 well plates This only changes the collection order The numbering of the wells remains as indicated on the plate The four virtual 96 well plates start at locations Al A13 I1 and 113 respectively The filling order of each quarter is as specified in the Collection Order section When the 384 well plate is split into four equal quarters the order of the four plates is the same as displayed in the Installed Plates section In the Reserved Locations you have the possibility to specify locations that will not be used for Fraction Collection see Table 8 Table 8 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
108. ting modes Time slices Peak threshold up downslope Timetable combination of time intervals and peak and Manual trigger supported only with Agilent Instant Pilot G4208A Agilent 1260 Infinity DAD MWD detectors G1315C D G4212B G1365 C D the Agilent 1260 Infinity Fluorescence Detector G1321A and the Agilent G1946C D G1956A B LC MSD are fully supported other detectors can be used but are not supported for fraction collection Discrete fractions default mode for all vessels The flow is diverted to waste while moving from one vessel position to the next vessel position Continuous flow optional available only when using the deep well plates It is possible to move from one well plate position to the next one without diverting the flow into the well plate to waste Needle into location Needle pushes into the vessel as deep as specified for the use with capped vials and test tubes and well plates with closing mats Droplet setup mode The tip of the fraction collector needle will initially move down to the bottom of the well Then it will slowly move upwards while the fraction is collected The droplet setup mode enables the fraction collector to collect small fractions without bubbles Agilent 1260 Infinity Fraction Collectors User Manual Specifications 6 Table 36 Performance Specifications Agilent 1260 Infinity Analytical scale Fraction Collector G1364C continued Type Specification Fraction vessel
109. tion 96 inlet waste tubings 105 inner capillary diameter 81 installing corrugated waste tubing 23 trays 33 waste tubings 33 installing the fraction collector 22 flow connections 31 interface cables 22 power cable 22 safety 22 installing the thermostatted fraction collector interface cables 26 power cable 26 preparation 26 safety 22 26 157 Index instrument status indicator 94 interfaces 143 146 internal tray 119 139 internet 155 into location 46 L liquid contact control 46 M maintenance functions 92 95 home position 96 park arm position 96 parts exchange position 95 transport unit self alignment 97 maximum capacity 143 146 flow rate 143 145 height 145 sizes 142 tube volume 142 145 method 47 setup 47 65 missing parts 14 Needle 136 needle 112 114 needle assemblies 136 needle movement 46 83 needle type 81 0 Online tick marks 75 operating modes 142 144 optimization 82 P park arm position 96 park transport unit assembly 41 parts and materials 14 125 158 accessory kit 140 closing mats 132 diverter valve assembly 137 internal tray 139 needle assemblies 136 plates 132 recommended tubes 128 recommended vials and caps 129 sheet metal kit 140 test tubes 128 transport unit assembly 134 135 trays 126 tubing kits 138 vials and caps 129 parts exchange position 95 peak based 66 peak detectors 66 performance speci
110. tive 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 Fraction Collectors User Manual Installing the Fraction Collector 1 Transporting the Fraction Collector When moving the Fraction Collector inside the laboratory no special precautions are needed However if the Fraction Collector 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 Fraction Collectors User Manual 41 1 42 Installing the Fraction Collector Transporting the Fraction Co
111. tton Agilent 1260 Infinity Fraction Collectors User Manual Configuration and Operation of the Fraction Collector 2 Viewing Your Results In order to display the tick marks for the collected fractions on the screen click Signal options from the Graphics menu Then choose Separated in the Layout dropdown list Report In order to create reports with a fraction table and tick marks the Specify Report box the item Add Fraction Table and Tick Marks has to be checked Specify Report Instrument 1 a File Type Area l MTT M WMF FDF csv F Xts ATM Figure 32 Fraction collection report setup Agilent 1260 Infinity Fraction Collectors User Manual 77 2 Configuration and Operation of the Fraction Collector Special Applications Pooling CAUTION We define Pooling as collecting fractions from multiple injections into the same fraction locations In the Agilent Chemstation there are two ways to initiate pooling e If you want to collect multiple injections from the one sample vial or sample well into the same fraction location specify multiple injections in the Sequence Table Incase you want to collect multiple injections from the several sample vials or sample wells into the same fraction location specify the same Fract Start for multiple samples in the Sequence Table An example is displayed in Figure 33 on page 78 In this sequence the Sample 1 will be injected from Plate 1 Position Al of the Agilent 1260 Inf
112. ues for threshold and slope The fraction collection is started if the detector signal exceeds both the threshold and the Up Slope value The fraction collection is stopped if the detector signal drops either below the threshold or the Down Slope value To specify the trigger values Up Slope Down Slope Threshold and Upper Threshold we recommend to use the Fraction Preview tool as described in Fraction Preview on page 70 Upper Threshold At high absorbance values the light intensity on the detector is extremely low and consequently detector noise will be superimposed on the detector signal In this case the detector noise might trigger fraction collection To avoid false fraction collection triggering we recommend setting an Upper Threshold well below the limit where this false triggering effect might occur As soon as the detector signal exceeds the Upper Threshold settings for Up Slope or Down Slope will be ignored until the signal drops again below the Upper Threshold 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 basing on the settings in the Peak Detectors table above Agilent 1260 Infinity Fraction Collectors User Manual 67 2 68 Configuration and Operation of the Fraction Collector If an MSD is used for mass based fraction collection Use MSD for mass based Fraction Collection must
113. unnel assembly through the slit beside the funnel in the internal tray assembly 6 Turn the internal tray upside down Slide the funnel s waste tubing through the flow delay sensor 7 Turn the internal tray upside down Slide the funnel s waste tubing through the flow delay sensor 122 Agilent 1260 Infinity Fraction Collectors User Manual Repairing the Fraction Collector 4 8 Insert the funnel into the tray and screw it tight clockwise 9 Re install the plug that holds the flow delay sensor 1 pull the waste tubing tight through the flow delay sensor 2 and mount the waste tubing 3 10 Re install the internal tray and make sure to route the waste tubing out through the outlet of the instrument correctly see Exchanging the Internal Tray on page 119 On completion of this procedure Re install the tray s in the tray base Start the instrument Close the front cover Agilent 1260 Infinity Fraction Collectors User Manual 123 4 Repairing the Fraction Collector Simple Repairs 124 Agilent 1260 Infinity Fraction Collectors User Manual Agilent 1260 Infinity Fraction Collectors User Manual 5 Parts and Materials Supported Trays for Fraction Collectors 126 List of Recommended Test Tubes 128 List of Recommended Vials and Caps 129 List of Recommended Plates and Closing Mats 132 Transport Unit Assembly Preparative Scale 134
114. ve Assembly preparative and analytical scale Item Description Part Number 1 Diverter valve assembly G1364 61901 2 PIN screw 0515 1211 Figure 45 Agilent 1260 Infinity Fraction Collectors User Manual Diverter Valve Assembly 137 5 Parts and Materials Tubing Kits 138 Table 32 Tubing Kits Item Description Part Number 1 Tubing kit preparative scale 0 8 mm ID consists of G1364 68711 items 2 and 3 2 Inlet waste tubing assembly preparative scale 0 8 mm ID Order Item 1 3 Diverter valve to needle tubing preparative scale 0 8 mm ID Order Item 1 4 Tubing kit analytical scale 0 25 mm ID consists of G1364 68712 items 5 and 6 5 Inlet waste tubing assembly analytical scale 0 25 mm ID Order Item 4 6 Injection valve to needle tubing analytical scale 0 25 mm ID Order Item 4 7 Low dispersion tubing kit analytical scale 0 15 mm ID 61364 68723 for flow rates lt 1 ml min 8 Inlet waste tubing assembly analytical scale 0 15 mm ID Order Item 7 9 injection valve to needle tubing analytical scale 0 15 mm ID Order Item 7 10 Finger tight fitting pack of 2 0100 1516 SI 10 2 5 8 3 6 9 Figure 46 Tubing Kits Agilent 1260 Infinity Fraction Collectors User Manual Parts and Materials 5 Internal Tray Assembly Table 33 Internal Tray Assembly Analytical and Preparative Scale Item Description Part Number 1 Internal tray
115. well plates only 2 x well plates std tray MTP for use with deep well plates only 10 x 2 ml vials 1 half tray 100 x 2 ml in std tray 1 half tray 3 x 40 x 2 ml in half tray 3x 15x 6 ml in half tray Full tray with 40 test tubes 30 mm OD max height 100 mm 45 ml tube Full tray with 60 test tubes 25 mm OD max height 100 mm 25 ml tube Full tray with 126 test tubes 16 mm OD max height 100 mm 12 ml tube Full tray with 215 test tubes 12 mm OD max height 100 mm 7 ml tube Installed trays are automatically detected and identified For the with uncapped vials tests tubes and well plates only Minimum 48 mm to 100 mm maximum ca 45 ml Agilent 1260 Infinity Fraction Collectors User Manual Specifications 6 Table 35 performance Specifications Agilent 1260 Infinity Preparative scale Fraction Collector G1364B continued Type Specification Maximum flow rate Delay volumes ul Delay calibration sensor Diverter valve cooling maximum capacity GLP features Interfaces Safety features 100 ml min depending on viscosity and generated back pressure max 6 bar at the diverter valve Fraction collector inlet to diverter valve 500 typical depends on length of the tubing Diverter valve 15 Diverter valve to needle 110 Needle 5 Single wavelength absorbance detector working at 654 nm consisting of a LED and a photo diode 3 2 Diverter valve wi
116. wrench for counter holding connect the valve to needle tubing to the needle Viewed from the bottom 8 IMPORTANT After fixing the screw clip the tubing into the guide on the bottom of the needle carrier assembly It is absolutely vital that the tubing is installed as On completion of this procedure Re install the needle to the needle carrier assembly if you previously removed it Make sure to slide the needle all the way to the front of the needle carrier assembly clicks into position Re install the tray s in the tray base Start the instrument Close the front cover Agilent 1260 Infinity Fraction Collectors User Manual 111 4 Repairing the Fraction Collector Exchanging the Preparative Needle Assembly Frequency When the needle is leaky or visibly damaged When the needle is blocked or contaminated Tools required Wrench open end 4mm PN 8710 1534 supplied in accessory kit Wrench open end 1 4 5 16 inch PN 8710 0510 supplied in accessory kit Parts required Preparative needle assembly G1364 87201 WARNING To avoid personal injury keep fingers away from the needle area during Fraction Collector operation CAUTION Regularly inspect the inlet waste tubing assembly and the valve to needle tubing and exchange them if they are worn out or show visible signs of damage Before beginning this procedure Position the transport unit of the Fraction
117. zation process At the end of this process the status LED should be green 24 Agilent 1260 Infinity Fraction Collectors User Manual Installing the Fraction Collector 1 CAN cable to previous module Control of Thermostat Remote CAN DC Out RS232C Iili uven DUNETAN aa AC Power CAN cable to next module Figure 7 Cable Connections The Fraction Collector 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 Fraction Collector from the line unplug the power cord The power supply still uses some power even if the switch at the front panel is turned OFF Agilent 1260 Infinity Fraction Collectors User Manual 25 1 Installing the Fraction Collector Installing a Thermostatted Fraction Collector Preparation Locate bench space Provide power connections Unpack the Fraction Collector 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 fro

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