Kontron Instruments HPLC GRADIENT FORMER 425 User manual

IA KONTRON
INSTRUMENTS
HPLC
GRADIENT
FORMER
Instruction manual

-K Instructionmanual GRADIENT FORMER 425 HPLC
TABLE OF CONTENTS
page
INSTALLATION
1.l Unpacking 2
1.2 Shipping list 2
1.3 Checking the serial number 3
*-~\4 Checking the mains supply 3
l.5 Installation 4
1.5.1Pump modification 4
1.5.2 Installing the gradientformer 5
1.5.3 Liquid connections 6
1.5.4 Electrical connection 6
1.5.5 Gas purge 7
1.5.6 Control Computer 7
2. INSTRUMENT DESCRIPTION
Generaldescription 2
2.1 Specifications 2
2.2 Front panel 3
2.2.1Controls 3
2.2.2Statusdisplay 3
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Instruction manual GRADIENT FORMER 425 HPLC
TABLE OF CONTENTS (cont.)
page
OPERATION
3.0 Programmable parameters 2
3.1 System start-up 2
3.2 Configuration 3
3.3 Main command modes 5
3.4 Manual mode 6
3.5 File programming 9
3.5.1Time 9
3.5.2 Flow 9
3.5.3 %A Step; %B Step; %C Step 10
3.5.4Sync 10
3.5.5 Auxiliary 11
3.5.6 A programmingexample 11
3.5.7 General points 13
3.5.8 Leaving edit mode 14
3.6 Saving and loading files 14
3.7 Running files 14
3.8 SystemStatus in file running mode 15
4. TROUBLE SHOOTING
4.1 Operationerrors 2
4.2 Softwareerror messages 3
5. MAINTENANCE
5.1 Routine maintenance 2
5.2 Spare parts 3
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13 Instruction manual GRADIENT FORMER 425 HPLC
TABLE OF CONTENTS
page
l. INSTALLATION
1.l ünpacking 2
1.2 Shipping list 2
1.3 Checking the serial number 3
1.4 Checking the mainssupply 3
1.5 Installation 4
1.5.1Pump modification 4
1.5.2 Installing the gradientformer 5
1.5.3 Liquidconnections 6
1.5.4 Electrical connection 6
1.5.5 Gas purge 7
1.5.6 Control Computer 7
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Ki Instruction manual GRADIENT FORMER 425 HPLC
TL. INSTALLATION
1.1 UNPACKING
Compare the consignment with the delivery note. Both accessories and
main unit are packed ina singlebox.Check the contents against the
following shipping l istand inform your local KONTRON representative
immediately should any parts be missing.
1.2 SHIPPING LIST
The following items are included äs Standard accessories:
1 91-00553 GradientFormer 425
2m 07-89023 Gas supply tube
1 55-65601 Mains cable
2 62-13001 Fuse 1.0A (110V)
2 62-13002 Fuse0.63 A (220V)
3 66-00452 Solventbottle
l 91-00560 Outlettube connection
6 92-00910 Inlet filter
l 92-00904 Inlet valve cartridge (titanium)
l 93-00667 Computercable
l 93-00668 Pump 420 cable
l 95-00196 InstructionmanualincludingSoftware
l 91-00568 Conversion kit, pump 420
l 52-69103 Connector, 4way
l 52-69104 Connector, 6 way
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Instruction manual
GRADIENT FORMER 425
HPLC
1.3 CHECK THE SERIAL HUMBER
The plate with the number is mounted onthe rear side ofthe Instrument.
This number must be identicalwith the serialnumber onthe delivery
note.
1.4 CHECKING THE MAINS SUPPLY
The GradientFormer 425is factory setfor 220V 50/60Hz. For Operation
with other mains voltages the correctvoltagemust besetonthe voltage
selector(see below) situated atthe rear ofthe Instrument.
Possible operating voltages: 100,120,220, 240 VAG
The fuses onthe voltageselectorhave tobe changed to correspondwith
the mainssupplyvoltage.
Frequency adjustment isnot necessary.
Voltage selector
- Unplugmains cable
- Open the snap-cover (A) using a
screwdriver atthe top.
- Pullmarker wheel (B)and turn it
to the requiredposition.
- Push (B) backintoposition.
- Pull out fuse holders (G) and
insert the correctfuse.
- Pushholders (C) backintoposi-
tion, obeyarrows.
- Close snap-cover(A).
- Check ifthe correctvoltage is
displayed.
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Instruction manualGRADIENTFORMER
425
HPLC
1.5 INSTALLATION
The GradientFormer 425isan accessory forthe KONTRONHPLCpump 420.
It has beendeveloped to operate with the medium head for thispump
(head part no. 91-00504).
A conversion kitis included inthe accessory kit. This kitis used to
convert a normalpump 420for Operationwith the gradientformer.
The Installation site ofthe gradient system should be suchthat itis
not exposed to dust or corrosivevapors. The ambient temperatureshould
be between 10°C and 35°C.
Allow at least 2 cm space around the System.
1.5.1Pump Modification
The pump 420 must be modified äs described inthe Installation instruc-
tion 84-00261 enclosedwith the conversion kit.
The pump 420 mustalso be fitted with a mediumpumphead(part no.
91-00504).
The connectionbetween the gradientformer andthe pumpmust be made
with pressure-proof connections. Itis thereforenecessary to replace
the inlet valve cartridge inthe pumphead.
Fig. 1.5.1 Pump 42O
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Ki Instruction manual GRADIENTFORMER 425 HPLC
1.5.1 Pump Modification (cont.)
The cartridge is replaced äs follows:
NOTE: pump 420 fitted with the inertpump kit Optionarticle no.
91-00554, havealreadybeenfittedwith the correct inlet
valve cartridge.
Remove the originalfitting by unscrewing the retaining nut shown
at (A)in fig. 1.5.1.
Remove the originalcartridge.
Insert the cartridgesuppliedwith the gradient former into the
retainingnut.
IMPORTANT: The black plastic sealing surface must faceinto the
pump head.
Screw the retaining nut backinto the pumphead,finger tight.
Using a Spannertighten one füll turn (approx. 80 cmkp).
1.5.2Installing the Gradient Former
Prior tothe initial Installation the following Instrument checks must
be made.
- Remove thetop cover(remove the fourfixing screws onthe base of
the unit).
- Check all connections tothe 3-waysolenoidvalvemanifold for
tightness.
- Check allgas connections, ensure that the supply tubes are pushed
äs far äs possibleinto the fitting.
— Replace thetop cover.
The GradientFormer 425 should be installed below the420 HPLCpump. The
solventbottlesshould be placed ontopofthe pump.
NOTE: It is recommended that the solventbottles are placed inthe420
System organiseraccessory for safer storage(notsupplied,part
no. 91-00530).
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InstructionmanualGRADIENTFORMER
425
HPLC
1.5.3Liquid Gonnections
GradientFormer - SolventReservoir
Pusheachsolventsupply andgas purging tube through the appropriate
hole inthe solventbottlecap.Note that the tubes are organised äs
three pairs, l and11,2 and22,3 and33. These should be kept
together.
Push oneofthe filterssuppliedinto theendofeach tube and place the
tube inthe appropriatesamplebottle.
The connectingtubes can be primed initiallyusing the appropriate purge
button anda syringe attached totheendofthe pump connection tube.
Gradient Former - Pump 420
The outlettube connection (part no. 91-00560) supplied1 with the
gradient formershould be connected between the420 pumpconnection on
the gradientformer andthe pump inlet fitting.
CAUTION: The fittings must notbe over-tightened. Usea second
Spanner to hold the retainingnut.
1.5.4Electrical Connections
Connect both pump and gradientformer toa suitable mains power supply
and switch on both units.
Gas Purge
Inlet
Connect ion
gradient former
pump 420
Connection
gradient former -
IBM-compatible PC
11.86
Fig. 1.5.4 ElectricalConnection - Gradient Former
l INSTALLATION
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Instruction manual GRADIENT FORMER 425 HPLC
1.5.4 Electrical Connections (cont.)
Connect the pump 420 connection on the gradient former with theRS232
connection onthe pump 420.Connect the Computerconnection onthe
gradientformerwith the COM-1 onthe IBM-compatible personal Computer.
Connect the SYNCinput onthe gradientformerwith the remote/continue
signal onthe460 autosampler if required. A detaileddescription ofthe
"sync" function is given in section 3 of this manual.
Connect the requiredauxiliaries for eontrol ofthe autosampler or other
HPLC Systemcomponents if required. A detaileddescription of auxiliary
connections and Programming is given is section 3 of thismanual.
1.5.5 Gas Purge
The gradientformeroperates onthe low pressure inlet tothe pump 420.
The solvent mixing occurs inthe inlettube and pump head itself. Degas-
sing during mixing can therefore occur and interfere with correct Opera-
tion ofthe System.This can generally be avoided by purging the sol-
vents withinert gas prior to starting.
The gradientformer is fittedwith three independent regulating valves
for controlling gas flow tothe differentsolvent reservoirs.
A suitableinert gas connection must therefore be made tothe gradient
former.Push the supplytubesuppliedwith the gradientformer firmly
into thegas purgeinletconnection(seefig. 1.5.4). Connectthistube
to an inert gas supply ata pressure not exceeding 0.5 bar.
W A R N I N G : The System isnot designed to operatewith pressurized
solvent storagebottles. The bottlesmust always be
installed such that no pressure build-up can occur.
The maximum inletpressure forthe purge gas must not
exceed O.5 bar.
1.5.6 Control Computer
The gradientSystem is controlled viaanIBM(or compatible) personal
Computer. ThisComputermust be installed äs described inthe handbook
supplied with the Computer.
The gradientformerSystemrequires a singlefloppydisk drive andone
RS 232C interface slot. No further hardware is required,auxiliary swit-
ching etcis carried out within the gradientformer.
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S Instruction manual GRADIENT FORMER 425 HPLC
TABLE OF CONTENTS
page
2. INSTRUMENTDESCRIPTION
General description 2
2.1 Specifications 2
2.2 Front panel 3
2.2.1 Controls 3
2.2.2Status display 3
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Instruction manual
GRADIENT FORMER 425
HPLC
2. GENERAL DESCRIPTIQN
The GradientFormer 425isan accessory tothe KONTRON HPLC pump420. It
allows formation of binary and ternary gradients onthe low pressure
side ofthe pump and thus requires only a singlehighpressureHPLC pump
in the gradientSystem.
The unit is designed to operate with a personalComputer äs SystemCon-
troller andis supplied complete with connection cables, a gas purging
System and solventreservoirs.
/~\1 SPECIFICATIONS
Solvent storage:
Gas purge:
Gradient capability:
Gradient increment:
Compositional accuracy:
Compositional precision:
3x11 glass bottles
3 independentregulatingvalves for
control of inert gas purge, max.
inlet pressure 0.5,bar.
Linear or stepgradientsfromeither
2or3 solvents.
Typically H- 1.5% measured with
water/water + 0.5% acetone mixtures
and 10%B stepgradients at1.5
ml/min.
Typically HH 0.4%measuredover 10
runs fora stepgradientfrom 0%to
50%B, conditions and flow äs above.
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K< Instruction manual
GRADIENTFORMER 425
HPLC
2.2 FRONT PANEL
Power
ON/OFF
to pump 420
Purgebutton
Valve Status LED
Fig. 2.2 Gradient Former425,Front panel
Gas purge
regulating
valve
2.2.1Controls
Gradientcontrol and parameter input is carried outbythe controlling
Computer. The gradient former therefore contains only two control ele-
ments äs described below:
PurgeButton
Pressing oneofthe threepurgebuttons causes the corresponding sole-
noid valve tobe switched to its open position. It remains openonly äs
long äs the purgebutton is keptpressed.
Gas PurgeRegulation
The three gas purgeregulatingvalvescontrol the rate of flow of pur-
ging gas into the correspondingsolventreservoir.
2.2.2StatusDisplay
Three LEDsindicate the currentStatus of each ofthe threesolenoid
valves. A valve is switched to open when the corresponding LEDis lit.
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-S Instruction manual GRADIENTFORMER 425 HPLC
TABLE OF CONTENTS
page
3.0 Programmable parameters 2
3.1 System start-up 2
3.2 Configuration 3
3.3 Main command modes 5
3.4 Manual mode 6
3.5 File programming 9
3.5.1Time , 9
3.5.2 Flow 9
3.5.3 %A Step; %B Step; %C Step 10
3.5.4Sync 10
3.5.5 Auxiliary 11
3.5.6 A programmingexample 11
3.5.7 Generalpoints 13
3.5.8 Leaving edit mode 14
3.6 Saving and loading files 14
3.7 Runningfiles 14
3.8 Systemstatus in file runningmode 15
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K4 Instruction manual GRADIENT FORMER 425 HPLC
3.0 PROGRAMMÄßIG PARAMETERS
The GradientFormer 425 Software allows Computercontrol ofthe fol-
lowing functions:
- Total flow.
- Percentagecomposition of mixtures containing upto3 different
solvents.
- Synchron!zation of gradient start with an autosampler (e.g. KONTRON
HPLC Autosampler 460).
- Switching ofupto four potential-free relay contactclosures inthe
Gradient Former425.
Control of these functions is exercised viaa program file of type.PRG
which contains a setof instructionswith a correspondingexecution time
for eachinstruction.
3.1 SYSTEM START-UP
The gradient former and pump 420 should be installed äs described in
section l of this manual. The Signal cable betweengradientformer and
pump should be connected andthe controlcable tothe gradientformer
connected betweengradientformer and COM1 onanIBMor IBM-compatible
PC.
CAUTION: Beforeusing the Software supplied with the gradientformer
you must make at least one back-upcopy for your own use.
" Never workwith the masterdisksuppliedwith the unit.
Make copies of yourcontrolSoftwareusing the diskcopy command äs des-
cribed in yourComputerhandbook.
Using the format/s command in MS-DOS youcan format yourduplicatedisk
such that an autoexecbatchfile canbe createdwhichwillautomatically
load the GF425 Software at system start-up. Thisprocedure is described
in the operatinginstructions for yourComputer.
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Instruction manual
GRADIENT FORMER 425
HPLC
3.2 CONFIGURATION
Insert the copy of your master disk inthe appropriate drive and type
GF425 followed by (CR) or enter. The control Software will be loaded and
the following maskwill be displayed.
KONTRON HPLC TERNARY GRADIENTCONTROLLER 425
Vers. 1.11
File name:
l ime
[min]
Repeats l ofl l Pressure. Upper-. 300 Lower:
Flow
[ml]
A Step
mm
B Step C
m i%] [*)
INFORMATION
Step Syn
Auxiliary [sec]
1234
Configuration cannot be read, please re-configure
Continue,
Fig. 3.2.1 Start menu without Configuration
The configuration cannot be readbecause no CONFHPLC file exists onthe
workingdisk.Thiswill be automaticallycreated atthe firstconfigu-
ration.
Press "C"to continue. The screenwillthenchange to display the normal
startingmask shown below:
KONTRON HPLC TERNARYGRADIENTCONTROLLER 425
Vers. 1.11
File name:
Repeats l ofl
Pressure, Upper: 200 Lower:
Time
[min]
Flow
[ml]
A Step
W (%]
B Step
(*] (%]
Step Syn Auxiliary [sec]
1234
SELECTED MODE: MAIN OPTION MENU
MODES: Run. Manual. Edit. Load. Save. Dir, Config. Quit.
3.2.2NormalStartingMenu
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Instruction manual
GRADIENT FORMER 425
HPLC
3.2 CONFIGURATIQN (cont.)
The system must be configured before proceeding. Forthe gradient former
the configuration file is used to define the upper and lowerpressure
limits for the pump 420, whether the pump should be switched offatthe
end ofa file batch andthecam correction factor. Select "C" to obtain
the configuration menu shown below.
KONTRON HPLC TERNARY GRADIENT CONTROLLER 425
Vers. 1.11
File name:
PUMP TYPE
HE AD
PRESSURE LIMITS Upper:
Lower:
PUMP OFFATENDOF BATCH (Y/N)?
CAM CORRECTIONFactor:
420
Medium
200
0
Y
5
SELECTED MODE: CONFIGURATION
MODES: ESC
Set pressure limit (0- 400 bar): 200
Fig. 3.2.3ConfigurationMenu
The cam correctionfactor issettoa default value of5 andis used to
make small corrections tothe linearity ofthe gradientprofile obtai-
ned. It is strongly recommended that this factor isnot changed. It has
no effect on reproducibility andis primarily intended äs a correction
factor for serviceuse.
NOTE:
You should only therefore usethe configuration module for
definition of systempressurelimits.
Set the requiredpressurelimits by moving the cursor into the corres-
ponding field and entering the valuerequired. The valueentered
together with the ränge of permitted entries is shown onthe last line
in all cases. The entry canbe completed by eitherpressingRETURN or
leaving the entryfieldusing the Cursor keys.
e.g. to enter an upper limit of250bar proceed äs follows:
- Move the cursor to upperlimit.
- Enter 250and pressRETURN.
-The value of250 will be displayed äs the upperlimit.
"Pump offatendof batch (Y/N)" is used to definewhether the pump
should be switched offattheendofa batch of program files or not.
Enter Y ifthe pump should be switched offandN ifthe pump should
continue atthe last set flowrate.
Once you have setthe requiredpressurelimitspressESCape to leave the
configurationmenu. The CONFHPLC.DAT filewill be automatically created
on yourworkingcopy.
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K Instructionmanual
GRADIENT FORMER 425
HPLC
3.3 MAIN COMMAND MQDES
A brief description ofthe main command modes availablefrom the main
starting menu is given below.
RUN:
is used to start a program file ora batch ofthe same file.
MANUAL:
allows execution of manually entered instructions.
EDIT: is used to create or modify program files.
LOAD: is used to load a previouslycreatedprogramfilefromdisk.
SAVE: is used to störe a previouslycreated or modifiedfile on
disk.
DIR:
gives a directorylisting ofall program(PRG)typefiles
stored onthe working disk.
CONFIG:selects the configuration menu.
QUIT: returns youto MS-DOS
Selection ofany command is made by entering the firstletter ofits
name, e.g. R for RUN.
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Instruction manualGRADIENTFORMER
425
HPLC
3.4 MANUAL MODE
Select "M" from the Starter menu to entermanual mode Operation,
following menuwill be offered:
The
KONTRON HPLC
TERNARY GRADIENT CONTROLLER 425
Vers. 1.11
File neune:
Repeats l ofl
Pressure. Upper: 200 Lower;
Time
[min]
0,0
Flow
[ml]
0.0
A Step
0
B Step
0
Step Syn Auxiliary [sec]
1234
Duration
[min]
0.0
0.0
Flow ABC
[ml ] [%] [%] [%]
0.0 1000 0
0.0 1000 0
Auxiliary [sec]
1
0
0
2
0
0
3
0
0
4
0
0
SELECTED MODE: MANUAL
COMMANDS: ESC. Execute. Aux only. Stop flow, Interrupt,
Duration (0..9999 min): 0.0
Fig. 3.4.1 ManualOperationMenu
All entries made in manualOperation are entered onthe bottom line of
the window.
The top line of this window isthe status line and displays the current
values forthe various parameter.
e.g. the duration field always shows the elapsed time since the last
instruction orthe last set value of duration ifthe gradient has
been completed. It is updatedevery 0.1 minutes.
CAUTION: The statusline issetto 1009SA äs default when manual mode
is first selected.Before starting your pump ensure that the
correctsolventcomposition for your column has been selec—
ted äs described inthe following example.
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Instruction manual GRADIENT FORMER 425 HPLC
3.4 MANUAL MODE (cont.)
Example l, ManualModeOperation
Assuming you wish toset your starting conditions to 80%A and 20%B ata
flow ofl ml/min.
- Move the Cursor (if necessary) to duration.
- Enter a time of0, pressRETURN (or move Cursor right).
- Enter a flow of l, pressRETURN.
- Enter 80for %A, pressRETURN.
- Enter 20for %B, pressRETURN.
- Press "E"to executethis instruction.
The Status line will change to display a flow of 1.0, 80%A and 20%B.
Example 2, ManualModeOperation
Using example l äs startingconditions and assuming you wish toruna
linear gradient from 0%C to 80%C with a constant 20%B component ina
time of10 minutes.
- Move the Cursor (if necessary) to duration.
- Enter a time of10 minutes,pressRETURN.
~ PressRETURN to enter the default value ofl forthe flow.
- Enter 0 for %A, pressRETURN.
- PressRETURN to enter the defaultvalue of20for %B.
- Enter 80for %C, pressRETURN.
- Enter "E"to executethisinstruction and start the gradient.
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