Bio-rad BioLogic LP System Manual de usuario

INSTRUCTION MANUALCatalog Numbers731-8300731-8301BIOLOGIC LPCHROMATOGRAPHY SYSTEM

1.3 UNPACKINGWhen you receive the BioLogic LP System, carefully inspect the shipping containers for any damage whichmay have occurred in shipping. Se

93SpecificationsValve Control• Proportioning valve/Mixer module, Diverter valve (SV-3), Bypass valve (SV-3), and Buffer Selectvalve (SV-5, or SV-3)• O

94Warranty and Ordering InformationAPPENDIX B. WARRANTY AND ORDERING INFORMATIONThe BioLogic LP System is warranted for 1 year against defects in mate

ORDERING INFORMATIONBioLogic LP Systems731-8300 BioLogic LP, 110VLP Controller, System Rack, UV Optics Module, Conductivity flow cell, Injection valve

Ordering Information (Continued)Peripherals731-8122 Model 2110 Fraction Collector, 110 V731-8120 Model 2110 Fraction Collector, 220 V731-8123 Model 21

Ordering Information (Continued)Cabling (continued)731-8267 System Cable 7, 8-pin mini-DIN to bare wires, connects the BioLogic LP to non-Bio-Radcompo

Ordering Information (Continued)BioLogic LP System Tubing731-8210 Silicone, 0.8 mm ID, 0.8 mm wall, 10 m length731-8211 Silicone, 1.6 mm ID, 0.8 mm wa

Life ScienceGroupWeb site www.bio-rad.com Bio-Rad Laboratories Main Office 2000 Alfred Nobel Drive, Hercules, CA 94547, Ph. (510) 741-1000, Fx. (510)

4IntroductionTable 1. (continued)Description of System ComponentsComponent FunctionSystem Rack This is the system organizer for columns and cartridges

5IntroductionTable 1. (continued)Description of System ComponentsComponent FunctionChart recorder Bio-Rad offers the Model 1327 Chart Recorder as a sy

6Description of Components2.0 DESCRIPTION OF COMPONENTSThe following sections identify the key features of each of the major system components.2.1 BIO

7Description of ComponentsTable 3. BioLogic LP Controller’s Rear Panel ConnectorsConnector DescriptionUV Optics: For connecting the UV Optics module

8Description of ComponentsTable 3. (continued)BioLogic LP Controller’s Rear Panel ConnectorsConnector DescriptionFraction Collector: The BioLogic LP

9Description of Components2.2 PROPORTIONING VALVE/MIXER MODULEThis is a combination proportioning valve and mixer. The solenoid valve proportions the

10Description of Components2.3 UV DETECTORThe UV Detector, which consists of the UV Optics Module and the control circuitry within the BioLogic LPCont

Figure 3 shows how the flow cell and the filters can be changed. To remove the flow cell, you will need toremove the retaining clip from the bottom o

2.5 BIO-RAD SYSTEM OPTIONSBio-Rad offers a number of system options for use with your BioLogic LP. The following instrument optionsare described in t

Table of ContentsTABLE OF CONTENTSSafetyGetting Started QuicklyChapter 1.0 Introduction...

2.5.2 Model 2128 Fraction CollectorThe Model 2128 provides X/Y motion drop-dispensing, accommodating a wide range of tube diameters andlengths, microt

2.5.3 Model 1327 Chart RecorderWhen used with the BioLogic LP, the Model 1327 Chart Recorder automatically starts when a methodbegins and stops when t

2.6 NON-BIO-RAD SYSTEM OPTIONSNon-Bio-Rad fraction collectors and chart recorders may be used with the BioLogic LP, as discussed in thefollowing secti

If you are using a non-Bio-Rad fraction collector, use an SV-3 diverter valve to enable advanced featuressuch as collection by Threshold and/or Collec

3.0 SYSTEM CONNECTIONSWhen setting up the system, consider the amount of bench space available, the instruments to be used withthe system, and the dev

To set up the BioLogic LP System: 1. BioLogic LP Controller.a. Place the BioLogic LP Controller on the bench and turn the unit so that the back side f

c. Insert the two short rods into the remaining holes. d. Mount the 2-piece column clamping arrangement across the two long rods using the attachedthu

Figure 11. System Cable Connections20System ConnectionsCONTROLLERMODEL 2110FRACTION COLLECTORMODEL 2128FRACTION COLLECTORORMODEL 1327CHART RECORDERSY

6. Chart Recorder; Model 1327.The Model 1327 chart recorder may be positioned on the rack shelf or on the bench. a. Connect System Cable 2 between the

4.0 SYSTEM PLUMBINGThis chapter begins with discussion of plumbing practices and general guidelines, followed by a general pro-cedure for plumbing the

Table of Contents5.3.5.3 Collection Windows Mode...675.3.5.4 Threshold and Collection

All plumbing connections require Luer fittings. A fittings kit is included with the BioLogic LP system. The fig-ure below shows the different Luer c

4.2 ADJUSTING THE PLATEN AND PLUMBING THE PERISTALTIC PUMPThe peristaltic pump requires the following tubing lengths:• PharMed 179 ±1.5mm• Silicone 17

2. Pull the platen cam lever away from the pump head to unlock the platen and slide the platen away fromthe pump head frame assembly, exposing the rol

4.3 PLUMBING THE SYSTEM1. Plumbing the inlets to the Proportioning valve/Mixer module.a. Cut two suitable lengths of Tygon tubing (0.8mm ID or 1.6mm I

Figure 14. System Plumbing27System PlumbingCOLUMNABCE DMV-6INJECTVALVESV-3COLUMN BYPASSVALVESAMPLE LOOPSV-3DIVERTERVALVEBUFFERSB AND APROPORTIONINGVA

6. Plumbing the MV-6 Valve’s Sample Loop.Plumb the inject valve’s sample loop according to Figure 14. Use male luer fittings at each end of theloop t

9. Plumbing the UV Optics Module and Conductivity Flow Cells.a. Connect the column outlet line to the inlet port of the UV Optics Module. (Arrows on

c. Using an SV-3 Diverter valve: Mount the valve on the System Rack so that it is as close as possi-ble to the Model 2128 Fraction Collector. Plumb

5.0 SYSTEM OPERATION5.1 FRONT PANEL CONTROLSTable 5. Front Panel ControlsKey DescriptionSoftkeys: These four keys are located directly below the LC

32System Operation!Table 5. (continued)Front Panel ControlsKey DescriptionMark key: This key signals the chart recorder to put an event mark on the

22. List of Methods ...522

33System OperationTable 5. (continued)Front Panel ControlsKey DescriptionInstr Instr (Instrument) keys: These keys allow you to select individual in

5.2 MANUAL MODEThere are three modes of operation: Manual mode, Program mode, and Run mode. Manual mode is usedto prepare the system to run a program

35System OperationMethod: <<name>> Manual OperationFlow: 1.00 ml/min UV: 0.0111 AUBuffer: A Cond: 0.000 mSSTART PURGE FLOW BUFFERMetho

Table 6. (continued)Manual Mode Operation: PumpReference DescriptionStart/Stop Screen: The first softkey in the Main Menu screenallows you to start/

37System OperationTable 6. (continued)Manual Mode Operation: PumpReference DescriptionCalibration for standard tubing sizes (0.8 mm, 1.6 mm, 3.2mm):

38System OperationTable 6.Manual Mode Operation: PumpReference DescriptionBuffer screen: Selecting Buffer from the Main Menu screendisplays the Buff

39System OperationTable 7. (continued)Manual Mode Operation: Fraction CollectorReference DescriptionStart/Stop screen: This first softkey in the Mai

Table 7. (continued)Manual Mode Operation: Fraction CollectorReference DescriptionF/X Size screen: This allows you to set the fraction size basedon

5.2.3 Manual Mode Operation of the AlarmsTable 8 discusses Alarm’s Manual mode functions. Figure 17 shows an overview of these functions.Table 8.Manu

5.2.4 Manual Mode Operation of the UV MonitorTable 9 discusses each of the functions available in Manual mode operation of the UV Monitor. For anover

SafetySAFETYCaution/WarningDisconnect power to any BioLogic LP component before servicing. No user-serviceable parts are inside any component. Refer s

43System OperationTable 9. (continued)Manual Mode Operation: UV MonitorReference DescriptionSet Max: Use this function to set a UV range other than

44System Operation5.2.5 Manual Mode Operation of the Conductivity MonitorTable 10 discusses the Conductivity Monitor’s Manual mode operation. Figure

Table 10. (continued)Manual Mode Operation: Conductivity MonitorReference DescriptionMin/Max: Allows you to set a range, from 0.00 mS to 999.99mS,

46System Operation5.2.6 Manual Mode Operation of the ValvesTable 11 discusses each of the functions available in Manual mode operation of the Valves.

47System OperationTable 11. (continued)Manual Mode Operation: ValvesReference DescriptionDivert screen: Allows you to set the SV-3 Diverter valve i

48System Operation5.2.7 Manual Mode Operation of the Chart RecorderTable 12 discusses each of the functions available in Manual mode operation of the

49System Operation5.3 PROGRAMMING MODEProgramming mode is used to define a sequence of actions which collectively comprise a method. Once amethod is

50System OperationFigure 21. Contents of a MethodMethod Name and Duration (Time or Volume) (Refer to section 5.3.2, Creating a New Method)PUMP TABLE(

51System Operation5.3.1 Program Mode’s Main MenuTo enter Program mode, press the Prog key. The Program display shows the currently opened method, and

52System OperationTable 13. (continued)Program Mode Operation: List of MethodsReference Description• Rename: This allows you to change the method’s

GETTING STARTED QUICKLYThis manual provides detailed discussion of the use and maintenance of the BioLogic LP system. Listedbelow is an overview of t

5.3.2 Creating a New MethodTo create a new method enter Programming mode by pressing the Program mode key. From theProgramming mode main menu the New

54System OperationTable 13. (continued)Program Mode Operation: New MethodReference Descriptionappropriate instrument key at any time, except while a

55System OperationMethod: <<name>> Volume: 7.00 mlLIST OF NEW SAVE VIEWMETHODS METHOD METHOD METHODSEE SECTIONS5.3.5 AND 5.3.6SEE SECTION

5.3.3 Viewing and Editing a MethodWhenever a method is Opened, or whenever View Method is selected, the Method List is displayed. Fromthe Method List

Table 15. (continuedProgram Mode Operation: Viewing and Editing a MethodReference DescriptionSave As... screen: This screen provides a number of opt

5.3.4 Program Mode’s Pump TableThe Pump Table displays the contents of all pump steps within the method. This table can be viewed bypressing the Pump

Table 16. (continued)Program Mode’s Pump TableReference DescriptionSelect Buffer: The buffers available are indicated by the lettersA through E plus t

Table 16. (continued)Program Mode’s Pump TableReference DescriptionLinear Gradient: A linear gradient consists of a continuouschange in the buffer com

61System OperationMethod: Volume: 1.00 mlStep: 1 Buffers: G M [A] B C D E(GRAD=Linear Gradient, MIX=%B Mixture)GRAD MIX CANCEL OKMethod: Volume:

5.3.5 Program Mode’s Fraction Collector TableThe Fraction Collector Table displays the fraction collector parameters within the method. This table is

3. Make final adjustments. From the Mode keys select Manual, and from the Instrument keys selectPump. Start the pump, and with fluid flowing through

Table 17. (continued)Program Mode’s Fraction Collector TableReference DescriptionFraction Collection Summary: This screen shows the currentcollection

5.3.5.1 Collect All ModeCollect All mode uses a single fraction size to collect fractions throughout a method. The units used for thefraction size are

5.3.5.2 Threshold Collection ModeThreshold Collection mode allows you to only collect peaks by defining a threshold value in absorbanceunits (AU). Wh

66System OperationMethod: Volume: 2.00 mlTHRESHOLD MODE Frac size: 1.00 mlThresh: 2.0000 AU Bubble Filter: MediumEDIT OKMethod: Volume: 1.00 mlSelect

5.3.5.3 Collection Windows ModeCollection Windows mode allows you to specify periods called “windows” within the method when fractionswill be collecte

Table 20. (continued)Program Mode’s Fraction Collector Table: Collection WindowsReference DescriptionWindow Start and End: Enter the times or volumes

69System OperationFigure 28. Programming: the Fraction Collector’s Windows Mode(Screen displays are keyed to the discussion in Table 20)Window Sta

5.3.5.4 Threshold and Collection Windows Collection ModeThreshold plus Collection Windows mode is a combination of Threshold Collection Mode and Colle

Table 21. (continued)Program Mode’s Fraction Collector Table: Threshold with Collection WindowsReference DescriptionWindow Start and End: Enter the t

72System OperationFigure 29. Programming: the Fraction Collector’s Threshold with Collection Windows Mode(Screen displays are keyed to the discussio

1.0 INTRODUCTION1.1 OVERVIEWThe BioLogic LP low-pressure, gradient chromatography system is designed for the purification of proteins,peptides, and ot

73System Operation5.3.6 Program Mode’s Alarm TableThe Alarm Table displays the alarms programmed within the method. This table can be viewed by press

Table 22. (continued)Program Mode’s Alarm TableReference DescriptionAlarm Settings: Enter the time or volume for the alarm to occurusing the number ke

5.3.7 Entering Method NamesSince up to 50 methods can be stored in the BioLogic LP, method names are necessary to keep track of themethods stored. Me

5.4 RUN MODERun mode is used to carry out the sequence of actions which are programmed within the current method.The name of the current method which

Table 24. (continued)Run Mode: Starting a RunReference DescriptionMultiruns: This allows the method to be run the method from 1to 999 times. Enter t

5.4.1.1 Errors Which Prevent the Start of RunsThe errors listed below can prevent a method from being run.• A Required Valve is Missing: A valve which

Monitor Flow Cell and the Fraction Collector Drop Head. This is best determined empirically using asyringe. To do this, first make certain the tubin

5.4.2.1 Information Available During a RunThere are two screens of information available ring a run which are described in Table 25.Table 24.Run Mode:

5.4.2.2 Holding a RunRuns can be placed on Hold manually by pressing the Hold softkey on the Run Screen or they may occurautomatically when an Alarm i

5.4.2.3 Pausing a RunRuns can only be Paused by pressing the Pause softkey on the Run Screen they cannot be programmed tooccur.Table 26.Run Mode: Run

1.2 FEATURESThe BioLogic LP System provides the following features:• A space-saving modular and stackable design which minimizes the system footprint

5.4.2.4 Manual OverrideMany of the operations which the BioLogic LP instruments can perform in Manual mode can also be per-formed in Run mode. To acc

• Conductivity Monitor: The Conductivity Monitor can be controlled exactly like it is in Manual mode,except that the Flow Cell cannot be calibrated.•

6.0 MAINTENANCE AND TROUBLESHOOTINGThe BioLogic LP System requires very little maintenance to assure reliable operation. This chapter discuss-es stan

6.2.1 Nominal CalibrationNominal flow rate factors for the three standard tubing sizes (0.8, 1.6, and 3.2mm ID) are programmed intothe system at the f

6.3 FLUSHING/CLEANING VALVES, FLOW CELLS, AND FILTERS6.3.1 Rinsing Valves and Flow CellsWhen the BioLogic LP will not be used for more than a day or t

6.4 CARE OF THE PROPORTIONING VALVE AND MIXERNormally, the only maintenance required is to flush the Proportioning Valve with water. To do this, use

3. Remove the screws holding the optics module together and remove the bottom half of the case. 4. Pull the UV lamp out of its holder and unplug it f

6.6 TROUBLESHOOTINGListed below are some guidelines for troubleshooting the system:Unstable baseline. This may be caused by any of the following:• Bu

Gradient not as expected.1. Check that the buffers are prepared properly, and the correct method has been selected.2. Check the range setting on Gradi

APPENDIX A. SPECIFICATIONSBioLogic LP SystemPower 115 V ~ 5.3 A230 V ~ 3.5 A50 - 60 HzOperating temperature 2° to 40°C, ≤95% humidityConstruction Mate