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Agilent 1100 Series Fraction Collectors User’s Guide

Configuration and Operation of the Fraction Collector

Application Notes

More information about the 1100 Series Fraction Collectors and Purification
Systems are available from a series of application notes. Printed versions can
ordered form Agilent or pdf-files can be downloaded from the Agilent Website

http://www.chem.agilent.com

Table 4

Selected Agilent Technologies Application Notes

Title

Publication Number

An optimized system for analytical and preparative work

5988-9649EN

Recovery collection with the Agilent 1100 Series purification system

5988-9650EN

Solutions for preparative HPLC-Application Compendium

5988-9646EN

Innovative fraction collection with the Agilent 1100 Series purification
platform

5988-9250EN

Automated fraction re-analysis - does it really make sense?

5988-8653EN

Injection of high-concentration samples with the Agilent 1100 Series
purification system

5988-8654EN

Sophisticated peak-based fraction collection - working with up and
down slope

5988-7895EN

Strategies for purification of compounds from non-baseline separated
peaks

5988-7460EN

Method scale-up from analytical to preparative scale with the Agilent
1100 Series purification system PS

5988-6979EN

Peak-based fraction collection with the Agilent 1100 Series purification
system AS - Influence of delay volume on recovery

5988-5747EN

Summary of Contents for G1364B

Page 1: ...s1 Agilent 1100 Series Analytical and Preparative Scale Fraction Collectors User s Guide...

Page 2: ...es 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 softwa...

Page 3: ...ollector including guidelines to optimize the system and to avoid problems 2 Troubleshooting and Test Functions This chapter describes the modules built in troubleshooting and test functions 3 Repairi...

Page 4: ...4Agilent 1100 Series Fraction Collectors User s Guide...

Page 5: ...rforming a delay calibration with an UV detector 18 Performing a delay calibration with an MSD 22 Making Your Own Calibration Method 26 Setting up a Fraction Collector Method 27 Fraction Preview 32 Se...

Page 6: ...roubleshooting 54 3 Repairing the Fraction Collector 55 Introduction into Repairing the Fraction Collector 56 Simple Repairs 56 Cleaning the Fraction Collector 56 Overview of Main Repair Procedures 57...

Page 7: ...88 Needle Assemblies 89 Diverter Valve Assembly 90 Tubing Kits 91 Internal Tray Assembly 92 Fraction Collector Accessory Kit 93 5 Specifications 95 Performance Specifications for the Fraction Collect...

Page 8: ...8 Agilent 1100 Series Fraction Collectors User s Guide Contents Agilent Technologies on Internet 109 Index 111...

Page 9: ...Collector Configuration of the Fraction Collector 10 Delay volumes and delay calibration 15 Setting up a Fraction Collector Method 27 Setting up a Fraction Collection Sequence 34 Viewing your Results...

Page 10: ...e tray configuration can be chosen from a wide variety of different trays and set up here For a selection of supported trays see Supported Trays for Fraction Collectors on page 80 Tube volume ml Volum...

Page 11: ...e 18 on page 39 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...

Page 12: ...ing 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...

Page 13: ...n 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 collectio...

Page 14: ...ollector the delay volume parameter will be ignored 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...

Page 15: ...imes tD1 and tD2 can be calculated by dividing the delay volumes by the flow rate The delay volume VD2 is a system constant and is 23 l for the fraction collector AS and 120 l for the fraction collect...

Page 16: ...detector and is accounted for when the diverter valve is triggered Tables 1 and 2 show the internal signal delay times for different Peakwidth settings Figure 3 Chromatogram with peak start t0 and pea...

Page 17: ...al delay time This Peakwidth setting should then be used for the purification run Signal delay time max VD1 Flow rate NOTE We recommend to set the Peakwidth always to 0 01 for the DAD and MWD or to 0...

Page 18: ...ly 5 Switch to Diagnosis View if necessary 6 Open the AFC Delay Volume Calibration status window see Figure 4 from the Fraction Collector sub menu of the Maintenance menu Maintenance Fraction Collecto...

Page 19: ...ration 7 Click the Continue button The Info field gives the instruction to turn on the pumps see Figure 5 8 When you have turned on the pumps click the Continue button again to start the delay volume...

Page 20: ...chromatograms from the detector s and the fraction collector delay sensor The results table contains the name and serial number of the detector the current delay volume stored in the fraction collect...

Page 21: ...onfigured you can calibrate the delay volume for the first fraction collector only the delay volume calibration menu is accessible only for Fraction Collector 1 which is the fraction collector that ap...

Page 22: ...ytical scale or DGCALPS M preparative scale and adjust the flow in the main pump to the flow you will be using for the analysis 7 Set the flow of the make up pump to the flow rate you will be using fo...

Page 23: ...in 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 19 Cut off a sufficient piece of the suppl...

Page 24: ...ion of the Fraction Collector in the ChemStation on page 10 23 Press the Print button to get a hard copy of the report 24 Stop the splitter 25 Return to the Method and Run Control View and load the me...

Page 25: ...ry 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 suff...

Page 26: ...ow 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 th...

Page 27: ...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 Coll...

Page 28: ...in case that the signal does not reach the condition to cut the fraction as exhibited in Figure 11 The could be caused by tailing peaks or if the baseline is drifting during gradient runs Peak Detect...

Page 29: ...and Upper Threshold we recommend to use the Fraction Preview tool as described in Fraction Preview on page 32 Upper Threshold At high absorbance values the light intensity on the detector is extremely...

Page 30: ...erall 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 the More button Ti...

Page 31: ...location can be specified If as configured is selected the pre configured volume see Instrument Pre configured Wellplate Types is used This ensures that the location well vial or tube cannot be overf...

Page 32: ...lows 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 u...

Page 33: ...specified and the upper and lower threshold level can be set up The graphically specified values are automatically transferred to the Peak Detector Table NOTE The run time for a system with a fraction...

Page 34: ...ox in the RunControl menu in the Sequence Parameters Figure 14 dialog box or in the Sequence Table in the Sequence menu All settings made in the Sequence Table overrule the settings made in the Sequen...

Page 35: ...t 1100 Series Fraction Collectors User s Guide 35 Configuration and Operation of the Fraction Collector 1 Setting up a Fraction Collection Sequence Figure 14 Start Location in Sequence Parameters dial...

Page 36: ...ropdown list To review your chromatograms file information and a fraction list select the Data Analysis view and press the Fraction Task button as displayed in Figure 15 In order to display the tick m...

Page 37: ...Agilent 1100 Series Fraction Collectors User s Guide 37 Configuration and Operation of the Fraction Collector 1 Viewing your Results Figure 16 Fraction collection report setup...

Page 38: ...ion specify the same Fract Start for multiple samples in the Sequence Table An example is displayed inFigure 17 on page 38 In this sequence the Sample 1 will be injected from Plate 1 Position A1 of th...

Page 39: ...location will be assigned from the ChemStation In order to disable the recovery the tray configuration has to be changed Standard tray for two well plates 10 x funnels G1364 84502 In this configuratio...

Page 40: ...ssembly on page 68 and Exchanging the Internal Tray on page 74 In addition the wellplate adapter has to be removed for operation with the shorter needle In the ChemStation Software the new configurati...

Page 41: ...ollector 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 the standard 50 mm ne...

Page 42: ...ng fractions is at least 0 05 min Peak based fraction collection Set threshold and slope values such that length of fractions is at least 0 05 min Unresolved peaks can be separated using appropriate t...

Page 43: ...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 between fraction collection is ign...

Page 44: ...Series purification system 5988 9650EN Solutions for preparative HPLC Application Compendium 5988 9646EN Innovative fraction collection with the Agilent 1100 Series purification platform 5988 9250EN...

Page 45: ...eries Fraction Collectors User s Guide Agilent Technologies 2 Troubleshooting and Test Functions Status Indicators 47 Maintenance Functions 49 Maintenance Functions 49 Transport Unit Self Alignment 51...

Page 46: ...rs Service Manual G1364 90102 Maintenance Functions The maintenance functions position the transport unit and needle carrier assembly on certain positions for maintenance homing parking or delay calib...

Page 47: ...e fraction collector The lower left indicates the power supply status the upper right indicates the fraction collector status Power Supply Indicator The power supply indicator is integrated into the m...

Page 48: ...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...

Page 49: ...Station the fraction collector maintenance positions can be selected from the Maintenance menu in the Diagnosis view see Figure 21 In the Control Module the functions can be selected in the Test scree...

Page 50: ...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...

Page 51: ...y The sample transport self alignment is required after disassembling the module or when you exchange The transport unit The needle capillary carrier assembly The MTP main board WARNING The sample tra...

Page 52: ...e Fraction Collector Transport Alignment to start the automated procedure The Transport Alignment Procedure takes approximately 10 15 minutes 6 Set the 8 bit configuration switch to the default settin...

Page 53: ...firm 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 predefin...

Page 54: ...collector responds to each command Table 5 Step Commands Step Action Comments Needle Up Lifts the needle arm to the upper position Command also switches the diverter valve to waste if it is not alread...

Page 55: ...5 Agilent 1100 Series Fraction Collectors User s Guide Agilent Technologies 3 Repairing the Fraction Collector Cleaning the Fraction Collector 56 Overview of Main Repair Procedures 57 Simple Repairs 5...

Page 56: ...damp cloth from which liquid could drip into the fraction collector WARNING When opening capillary or tube fittings solvents may leak out Please observe appropriate safety procedures for example goggl...

Page 57: ...ytical scale module G1364C The preparative scale fraction collector G1364B contains a different needle Internal tray with rinse port and flow delay sensor Diverter Valve Transport unit Plug channel wi...

Page 58: ...Needle Tubing on page 63 Exchanging the preparative needle assembly When needle shows indication of damage or blockage See Exchanging the Preparative Needle Assembly on page 66 Exchanging the analyti...

Page 59: ...reparative scale 0 8 mm ID PN G1364 68711 or Inlet waste tubing assembly included in tubing kit analytical scale 0 25 mm ID PN G1364 68712 WARNING To avoid personal injury keep fingers away from the n...

Page 60: ...nt 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...

Page 61: ...flow cell of the detector The front tubing must be inserted into the waste container 6 Connect the finger tight fittings of the inlet waste tubing assembly to the ports of the diverter valve IMPORTAN...

Page 62: ...airing the Fraction Collector Simple Repairs 8 IMPORTANT The cables must run into the ports of the diverter valve in lines parallel to the horizon On completion of this procedure Re install the tray s...

Page 63: ...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 tub...

Page 64: ...assembly at the diverter valve 3 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 4 Un clip the tubing from the b...

Page 65: ...the 5 16 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 needl...

Page 66: ...peration 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...

Page 67: ...bing 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...

Page 68: ...AUTION 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...

Page 69: ...le 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 On co...

Page 70: ...70 Agilent 1100 Series Fraction Collectors User s Guide 3 Repairing the Fraction Collector Simple Repairs...

Page 71: ...sition see Maintenance Functions on page 49 Remove all installed trays from the tray base Position the transport unit of the fraction collector in the Change Parts Position see Maintenance Functions o...

Page 72: ...e from the connector on the front of the z arm 5 Using the 2 mm hex key loosen the holding screw of the diverter valve and remove the diverter valve from the z arm 6 Install the new diverter valve to...

Page 73: ...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...

Page 74: ...al scale G1364 63113 Internal tray preparative scale G1364 63114 Before beginning this procedure Position the transport unit of the fraction collector in the Home Position see Maintenance Functions on...

Page 75: ...of the outlet 2 the before sliding the tray out 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...

Page 76: ...of the fraction collector in the Home Position see Maintenance Functions on page 49 and remove all installed trays from the tray base Turn off the instrument The procedure is almost identical for both...

Page 77: ...embled 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 funnel assembly through the slit beside the funnel in the...

Page 78: ...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 th...

Page 79: ...s 80 List of Recommended Test Tubes 82 List of Recommended Vials and Caps 83 List of Recommended Plates and Closing Mats 85 Transport Unit Assembly Preparative Scale 87 Transport Unit Assembly Analyti...

Page 80: ...ates 10 2ml vials G1367 60001 7 Std tray for 2 well plates 10 collecting funnels G1364 84502 8 Funnel for item 7 5022 2200 9 Funnel seal kit for item 7 pack of 10 G1364 68730 10 Tubing kit 10T for ite...

Page 81: ...action Collectors User s Guide 81 Parts and Materials 4 Supported Trays for Fraction Collectors Figure 24 Trays 6 7 5 4 3 12 13 14 15 16 2 1 The items in this figure are displayed in different scaling...

Page 82: ...s and Materials List of Recommended Test Tubes List of Recommended Test Tubes Table 8 Round Bottom Test Tubes Description Volume ml lOO Pack 25 x 100 mm clear glass 35 5042 6459 30 x 100 mm clear glas...

Page 83: ...4493 5183 4495 Table 10 SnapTop Vials Caps for Use with the Analytical Scale Fraction Collector only Description Volume ml lOO Pack lOOO Pack lOO Pack silanized Clear glass 2 5182 0544 5183 4504 5183...

Page 84: ...Use with the Analytical Scale Fraction Collector only Description Septa 100 Pack Clear polypropyIene Clear PTFE red rubber 5182 0550 BIue polypropylene Clear PTFE red rubber 5182 3458 Green polypropyl...

Page 85: ...glass inserts caps and septa pre assembled 0 35 1 5065 4402 5 384 polypropylene well plate 0 1 30 5042 1388 6 96 polypropylene conical well plate 0 18 25 5042 8502 7 54 x 2ml vial plate 1 5 6 G2255 6...

Page 86: ...Mats 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...

Page 87: ...ion Part Number 1 Transport Unit Assembly Preparative Scale includes items 2 and 4 G1364 60018 2a b Tubing diverter valve to needle 0 8 mm ID G1364 86711 3 Diverter valve assembly G1364 61901 4 Needle...

Page 88: ...unit assembly analytical scale includes items 2 and 4 G1364 60019 2a b Tubing diverter valve to needle 0 25 mm ID G1364 68712 3 Diverter valve assembly G1364 61901 4 Needle carrier assembly analytica...

Page 89: ...Item Description Part Number 1 Needle assembly preparative scale G1364 87201 2 Needle assembly analytical scale 20 mm needle For use in the analytical scale fraction collector at flow rates 10 ml min...

Page 90: ...arts and Materials Diverter Valve Assembly Diverter Valve Assembly Table 19 Diverter Valve Assembly preparative and analytical scale Item Description Part Number 1 Diverter valve assembly G1364 61901...

Page 91: ...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 items 5 a...

Page 92: ...e includes items 2 7 Internal tray assembly preparative scale includes items 2 7 G1364 63114 G1364 63113 2 Screw seat adapter 5022 2200 3 Seal funnel reorder No pack of 10 G1364 68730 4a Waste Tubing...

Page 93: ...10 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 Reorder gives pack of 2 3 0100 1516 Waste tubing 1 2 m Reorder gives 5 m 1 5062 2463 CAN c...

Page 94: ...94 Agilent 1100 Series Fraction Collectors User s Guide 4 Parts and Materials Fraction Collector Accessory Kit...

Page 95: ...95 Agilent 1100 Series Fraction Collectors User s Guide Agilent Technologies 5 Specifications Performance Specifications for the Fraction Collectors 96...

Page 96: ...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 Fraction capacities and tray...

Page 97: ...60A GLP features Early maintenance feedback EMF electronic records of maintenance and errors Interfaces Controller area network CAN optional LAN or external contacts interface RS232C APG remote for re...

Page 98: ...tion 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...

Page 99: ...g of a LED and a photo diode diverter valve 3 2 Diverter valve with low internal volume 15 l switching time 100 ms maximum operating pressure 6 bar cooling Optional with additional G1330B performance...

Page 100: ...on Collectors Vials and well plates and capped vials and well plates with closing mats can be used as recom mended by Agilent Technologies see List of Recommended Vials and Caps on page 83 and List of...

Page 101: ...ion Collectors User s Guide Agilent Technologies A Safety Information Safety Information 102 Lithium Batteries Information 105 Radio Interference 106 Sound Emission 107 Solvent Information 108 Agilent...

Page 102: ...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...

Page 103: ...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 b...

Page 104: ...Indicates dangerous voltages Indicates a protected conductor terminal Eye damage may result from directly viewing the light produced by the Xenon flash lamp used in this product Always turn the xenon...

Page 105: ...egulated 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 WARNING Lithiumbatteri Ekspl...

Page 106: ...y 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...

Page 107: ...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...

Page 108: ...y 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 O2 2COCl2 2...

Page 109: ...TA ethylene diamine tetra acetic acid Mixtures of carbon tetrachloride with 2 propanol or THF Agilent Technologies on Internet For the latest information on products and services visit our worldwide w...

Page 110: ...110 Agilent 1100 Series Fraction Collectors User s Guide A Safety Information Agilent Technologies on Internet...

Page 111: ...verter valve 97 99 diverter valve assembly 90 E error messages 46 49 F failure 46 fill volume maximum 31 fraction capacities and trays 96 fraction delay volumes 12 fraction list 36 fraction preview 32...

Page 112: ...ngs 59 internal tray 74 main procedures overview 57 needle 66 68 preparative needle assembly 66 simple repairs 56 58 valve to needle tubings 63 report 36 reserved locations 13 rinsing 30 43 S safety f...

Page 113: ......

Page 114: ...ilent Technologies In This Book This manual contains technical reference information about the Agilent 1100 Series fraction collectors The manual describes the following configuration and operation of...

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