User'S Manual: Version 1.4, March 1995
User'S Manual: Version 1.4, March 1995
DECLARATION OF CONFORMITY
We
G. de Pee, QA manager
If you have any comments on this manual, we will be pleased to receive them at:
SPARK HOLLAND BV,
P.O.BOX 388,
7800 AJ EMMEN,
THE NETHERLANDS.
0060.103-14
SAFETY PRACTICES
General
The following safety practices are intended to insure the safe operation of the equipment.
Electrical Hazards
1. Removal of some panels exposes potentially dangerous voltages. Disconnect the instrument from all
power sources before removing protective panels.
2. Replace blown fuses with size and rating stipulated on the fuse panel or holder, and in the manual where
listed.
3. Replace or repair faulty or frayed insulation on power cords.
4. Check actual line voltage to confirm it is the value for which this instrument is wired. Be sure power
cords are plugged into correct voltage sources.
General Precautions
1. Perform periodic leak checks on supply lines.
2. Do not allow flammable and/or toxic solvents to accumulate. Follow a regulated, approved waste disposal
program. Never dispose of such products through the municipal sewage system.
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ii
SPECIFICATIONS
GENERAL
Sound pressure level
<70 dB
Working temperature
10 - 35 C
Installation category
Pollution degree
Injection volume
flushed loop:
5 - 500 l (dispenser controlled),
5 - > 5000 l (by headspace pressure).
partial loopfill:
(only Basic-plus)
10 - 490 l in steps of 10 l. (depending on the
loopvolume, maximal 50% of the loopvolume)
max. 9
Analysis time
Needle wash
Headspace pressure
ANALYTICAL PERFORMANCE
Reproducibility
Memory effect
(only Basic-plus)
ELECTRICAL
Power requirements
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iii
Inject marker
Stop I/O
Vial number BCD output
Auxiliary 1
Auxiliary 1 inverted
Auxiliary 2
Auxiliary 2 inverted
INPUTS
OPTIONS
Sample tray cooling
Micro injector
PC Control
PHYSICAL
0060.103-14
Dimensions ( W D H )
Weight
iv
CHAPTER 1 INTRODUCTION
The BASIC-MARATHON autosampler is dedicated to routine analysis of large series of samples in an
environment where zero sample loss and injection flexibility are of little significance but where reliability,
stability and reproducibility is mandatory. The BASIC -MARATHON therefore makes use of flushed loop injection,
guaranteeing reproducibility of injection better than 0.5 % and the Basic-plus has additional the partial loopfill
injections mode, guaranteeing reproducibility of injection better than 1.0%.
0060.103-14
1.1
1. Dispenser aspirates approximately 10 l air bubble in the sample needle for more efficient removal of
the previous sample.
2. With the injection valve in inject position, the coaxial needle pair is inserted into the sample. Air
pressure, applied to the sample through the outer air needle, ensures that no air or vapour bubbles are
formed during sample withdrawal.
3. Dispenser withdraws "preflush volume" from sample vial to fill the sample line with sample and to
remove previous sample. (30 - 999 l, programmable)
4. Injection valve switches into load position; dispenser transports 2 or 3 x loopvolume through the loop
to fill the loop quantitatively.
2 x loopvolume for loops equal to or larger than 100 l,
3 x loopvolume for loops smaller than 100 l.
In case of partial loopfill injections (Basic-plus) the loop is filled with the programmed injection volume.
5. Injection valve switches into inject position. Sample loop is now part of the HPLC mobile phase flow
path: sample is transported to the column. Analysis time starts.
If one injection is to be made out of each sample vial, the needle pair withdraws after analysis time and
sequence 1-5 starts anew for the next vial. If more than one injection is to be made out of the same sample
vial, the needles remain in the vial and after analysis time thenext injection is made starting the sequence at
3 with a fixed preflush of 4 l.
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1.2
1. With the injection valve in load position, the coaxial needle pair is inserted into the sample. Air
pressure, applied on the sample through the outer (air-) needle, will force the sample via the inner
(sample-) needle through the sample loop. The duration of the headspace pressure is programmable;
appropriate duration has to be determined experimentally.
2. Injection valve switches into inject position. Sample loop is now part of the HPLC mobile phase flow
path: sample is transported to the column. Analysis time starts.
A special sample drain, that prevents siphoning, must be connected to the injection valve (replacing the
dispenser connection), refer to section 4.2.3.3.
If one injection is to be made out of each sample vial, the needle pair withdraws after analysis time and
sequence 1-2 starts anew for the next vial. If more than one injection is to be made out of the same sample
vial, the needles remain in the vial.
1.3
Working principle
The DDS is a miniature "finger pump". The operating principle is shown in Figure 2.3: Fingers 1 and 3 act as
inlet and outlet pinch valve respectively, finger 2 provides the fluid displacement. The stroke of finger 2 is
factory adjusted to provide 2 l per stroke. Since the volume dispensed is always a multiple of one stroke
volume (digital dispensing), i.e. 2, 4, 6 l etc., odd values for volumes will be round down to even values.
0060.103-14
0060.103-14
GENERAL DESCRIPTION
1.Sample tray, consisting of 4 removable segments
2.
Keyboard
3.
Display
4.
Cover with hinges
5.
Needle arm with sample needle
6.
Injection valve with sample loop
7.
Dispenser cassete screw
8.
Dispenser
9.
Outlet tubing holder
10.Common leakage/condensation water drain for injection valve and sample tray.
1.5
INJECTION SYSTEM
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
17.
18.
19.
20.
0060.103-14
CHAPTER 4 INSTALLATION
1.6
UNPACKING
1.7
CONNECTIONS
3.
4.
5.
6.
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7.
8.
9.
10.
11.
Before plugging in the power cable, check voltage settings of the modules at the input socket on the rear
panel.
Make sure that the voltage settings of the modules are identical with the voltage of your local power
supply.
Use only a supply appliance with protective grounding.
If the indicated voltage is not correct, select the proper voltage by removing, inverting, and than reinserting the voltage selector.
When the voltage selection and fuses are correct for your power source, plug in the power cable.
RS232 connector
1.7.2.2
Auxiliaries connector
Pin No.
1
2
3
4
5
6-8
1.7.2.3
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I/O description
+ 24 volt DC
Auxiliary output 2 inverted
Auxiliary output 2
Auxiliary output 1 inverted
Auxiliary output 1
Ground
Pin No.
1
3
4
I/O description
Inject marker output (TTL, active low)
5Stop I/O (TTL, pulled up); output is low when BASIC-MARATHON ends processing (READY); pull low input will stop
9-15
Ground
1.7.2.4
8-bit BCD-coded VIAL NUMBER OUTPUT (True logic), representing vial numbers 1-96; active when
corresponding vial is processed.
Pin no
1
2
3
4
8
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Output
Bit 0 units 1
Bit 2 units 4
Bit 4 units x 10
Bit 6 units x 40
Ground
Pin no
9
10
11
12
15
Output
Bit 1 units 2
Bit 3 units 8
Bit 5 units x 20
Bit 7 units x 80
Inject marker (TTL, active low)
Value
Output
D0
(1)
l = ON
D1
(2)
D2
(4)
D3
(8)
0
l
2
l
l
l
4
l
6
l
l
l
8
l
Tenth
BCD
D7
D6
D5
D4
D3
D2
D1
D0
(8)
Output
Ones
(4)
(2)
(1)
(8)
9 10 (10 )
0
5 10 (10 )
0
1 10 (10 1)
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(1)
0 1 (100)
59
50 + 9
9 1 (10 )
0
96
90 + 6
Output
(2)
6 1 (10 )
Output
(4)
Vial number
10
10 + 0
1.7.3.1
Sample loop
The DDS pumptubing cassette is factory-installed and connected to the injection valve. Fasten the
pumptubing cassette before use with the screw on top of the pumptubing cassette. If the pumptubing
cassette needs replacement, refer to Appendix A.
1.7.3.3
TIME based loopfilling by sample headspace pressure can be selected (SYSTEM SETTINGS, ref. 5.5.1) to
shorten loopfill time for larger loop volumes (Ref. Chapter 2, INJECTION PRINCIPLE).
To allow time based loopfilling and to prevent hydrostatic sample loss, the special sample drain for
headspace pressure loopfill must be connected at the injection valve, replacing the dispenser connection at
the valve (Ref. 3.2 for description).
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10
1.8
PREPARATION OF VIALS
2-CV
11-AC4
(1.5 ml vial)
(crimpcap)
Spark p/no's:
For small sample volumes or for low sample loss (low rest volume, see 4.3.2) use Chromacol 1,1 ml conical
vials with support sleeves (ask your local SPARK dealer):
Chromacol ref. :
Spark p/no's :
3760.407
3760.408
3760.402
3760.409 (5 ml vial)
3760.410 (crimpcap)
If other vials are to be used, make sure that the following specifications are met:
minimum height:
33 mm
maximum height:
40 mm (including cap)
maximum diameter:
11.8 mm (standard tray)
minimum cap diameter:
10 mm
minimum space for septum penetration: 4 mm diameter (hole in cap)
The use of caps with a diameter smaller than 10 mm may cause damage to the BASIC-M ARATHON .
The use of Eppendorf microcentrifuge tubes may cause malfunctioning of the vial sensor of the
BASIC-MARATHON.
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11
Example: 3 flushed loop injections out of a standard sample vial using a 20 l loop and 30 l pre flush
volume, minimum sample volume needed is:
200l+30l+3x3x20l+ 2x4l=418l
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12
Check seal after crimping: if cap can be turned easily, the seal is not airtight (readjust handcrimper).
1.8.4 LOADING THE SAMPLE TRAY
To allow remote loading of the sample tray, the sample tray consists of four removable segments that fit on
one UNIQUE position of the tray holder only.
Use the STEP key (ref. 5.3, KEY FUNCTIONS) to rotate the tray (functioning only when the BASIC-M ARATHON
is in READY status).
DO NOT ROTATE THE TRAY MANUALLY;
YOU MAY DAMAGE THE TRAY DRIVE MECHANISM!
Positioning of vials
Sample vials should be placed in sequential positions on the tray. Empty spaces between vials of a sample
series are allowed, since the BASIC-MARATHON can detect empty positions. After detecting an empty place
the BASIC-MARATHON will continue with the next position.
Additional samples can be placed during RUN since LAST VIAL value can be changed during RUN (Ref. 5.6,
RUN).
Needle wash vials
If needle wash is programmed (ref. 5.5.1, SYSTEM SETTINGS), place 4 vials with washing solvent (1.5 ml
each, usually mobile phase) on positions 93-96 (place all 4 vials; they will be used alternately).
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13
CHAPTER 5 OPERATION
Operating the BASIC-MARATHON is easy; after programming the system SETTINGS, only a few parameters
need to be programmed for regular automatic processing of a series of samples.
All parameter values are saved in memory after power switch OFF, except tray temperature setting, if tray
cool option is installed: tray cooling is always OFF after power UP.
1.9
PRINCIPLE
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Selected mode
F,9
F,7
F,2
F,3
F,4
F,0
14
Key:
Function:
STEP key; used to step through the parameter list. The STEP key is also used to
rotate the sample tray (in READY status only!).
Hold/Continue countdown of analysis time. Analysis time is extended with the time
that hold is active.
Function key; used in combination with a numerical key to select or exit the
various modes.
Numerical keys; used to enter values for programmable parameters. Values are
stored when going to the next parameter (STEP) or when ending programming
(PROG/END).
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15
READY
Use
SYSTEM SETTINGS
to step.
1. FIRST VIAL
(1 48/92/96)
2. LAST VIAL
(1 48/92/96)
3. NO OF INJ'S/VIAL
(1 9)
4. PREFLUSH VOLUME (30 999l)
FLUSH TIME
(1 999SEC)
5. INJECTION VOLUME (10 490l)
6. ANALYSIS TIME
7.
8.
9.
10.
TIME
TIME
TIME
TIME
AUX
AUX
AUX
AUX
COOLING CONTROL
1
1
2
2
ON
OFF
ON
OFF
(0:00
(0:00
(0:00
(0:00
99:59)
99:59)
99:59)
99:59)
(0:00 99:59)
Use
to step.
1. LOOPVOLUME
(0999l)
2. SAMPLE VOLUME
(STD/LSV)
3. INJ. MODE (FLUSHED/PARTIAL)
4. FLUSH
(VOL/TIME)
5. NEEDLE WASH
(YES/NO)
6. AUXILIARIES
(YES/NO)
7. MARKER PULSE LENGTH (0.12.0)
PRESS
RUN MODE
NOTE
:
Only available in
the Basic-plus autosampler
ANALYSIS TIME
Accessible: Run parameters, Cooling control.
PRIORITY MODE
default vial no 00!
Accessible:
Run parameters,
Cooling control.
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16
1.13 PROGRAMMING
Only four modes contain programmable parameters:
-
1.13.1
SYSTEM SETTINGS
With BASIC-M ARATHON in READY status, key in F,9 to select SYSTEM SETTINGS mode. Press PROG/END to
enable programming of the following parameters (use STEP key to go to the next parameter):
- LOOP VOLUME;
- SAMPLE VOLUME (STD/LSV);
0060.103-14
- FLUSH (VOL/TIME);
- AUXILIARIES (YES/NO);
Enter the pulse length of the inject marker; 0.1 - 2.0 sec.
17
1.13.2
RUN PARAMETERS
With BASIC-M ARATHON in READY status, press PROG/END to select RUN PARAMETER mode and to enable
programming at the same time.
The following parameters can be programmed (use STEP key to go to next parameter):
- FIRST VIAL;
enter position number of the first vial of the sample series in the sample tray.
1-96 without needle wash,
1-92 with needle wash,
1-48 in case of LSV.
(ref. 5.5.1 SYSTEM SETTINGS)
- LAST VIAL;
enter position number of the last vial of the sample series in the tray.
(see FIRST VIAL for valid numbers)
- INJ/VIAL;
(parameter shows up for flush on volume basis only; ref. 5.5.1 SYSTEM
SETTINGS); enter volume (of sample) which is to be used to flush needle +
connecting tubing prior to filling the loop.
30-999 l.
- FLUSH TIME;
(parameter shows up for flush on time base only; ref. 5.5.1 SYSTEM SETTINGS);
enter duration of headspace pressure for loopfilling.
1-999 sec.
- INJECTION VOLUME;
(Basic-plus only)
Enter the injection volume for the partial loopfill injections.
10 - 490 l in steps of 10 l. (maximal 50 % of the loopvolume)
- ANALYSIS TIME;
enter the time interval needed before the next injection takes place.
0-99'59". Time countdown starts at moment of injection.
(parameter shows up only when auxiliaries are enabled in System settings (ref.
5.5.1). Enter time at which auxiliary 1 must switch ON.
0-99'59".
(parameter shows up only when auxiliaries are enabled in System settings (ref.
5.5.1). Enter time at which auxiliary 1 must switch OFF.
0-99'59".
see AUX 1.
0060.103-14
see AUX 1.
18
1.13.3
Sample tray temperature control is switched ON/OFF in the COOLING CONTROL mode.
With BASIC-MARATHON in READY status, key in F,7 to select COOLING CONTROL mode. Press PROG/END to
switch ON or OFF the cooling control.
The sample tray cooling has a fixed temperature of 4C
- TRAY COOLING (ON/OFF);
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19
1.14 RUN
When programming is ready (Ref. 5.5), system is ready (Ref. CHAPTER 4, INSTALLATION) and
BASIC-M ARATHON is in READY status, press START/STOP key to start automatic processing of the samples.
During RUN, BASIC -MARATHON will show information relevant to the actual process like: SEARCH VIAL 45,
LOOPFILL, WASHING etc. During analysis time the display will show the following information continuously
(example):
VIAL:15 82 2'INJ
4C
The first line shows that the second injection from vial 15 is being processed, vial 82 is the last vial to be
processed. The temperature of the cooling tray shown at the end of the first line will only be displayed if the
option is present.
The second line shows the time since the injection. The status of the auxiliaries, the number will be
displayed if the auxiliary is ON. (Only shown when enabled in the settings)
The first character of the second line will be a blinking r if the BASIC-MARATHON is under remote control or a
blinking p if a priority sample is programmed.
It is possible to reprogram RUN PARAMETERS during RUN; simply press PROG/END to enter the RUN
PARAMETER mode (ref. 5.5 for programming). Press PROG/END to exit RUN PARAMETER mode and to
continue the RUN program.
If you do not return to RUN mode, RUN will stop after finishing the current injection!
Changes in the RUN parameters will be executed immediately after returning to RUN mode (although an
injection will not be interrupted).
COOLING CONTROL can also be reprogrammed during RUN; press F,7 to enter COOLING CONTROL mode
(ref. 5.5 for programming).
Return to RUN (F,0) to continue RUN!
1.14.1
HOLD
HOLD (press HOLD/CONT) can be used to freeze the time base. During HOLD, the RUN PARAMETER mode,
COOLING CONTROL mode and PRIORITY mode are accessible to reprogram/check parameters (ref. 5 .5).
Exit any of these modes (if entered) and press HOLD/CONT key to continue RUN. Analysis time of the
current injection has then been extended with the time that HOLD was active.
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20
1.14.2
PRIORITY SAMPLE
A sample run can be interrupted to process an emergency sample prior to the next sample of the
programmed samples series: press F,2 to enter the priority mode, press PROG/END key and enter the
number of the tray position where the priority vial has been placed. Press PROG/END to end programming.
Return to RUN (F,0). Priority vial will be processed immediately after the current vial. A blinking "p" (first
position of lower display line) indicates that a priority vial is to be/being processed. After processing the
priority vial, RUN will continue where it was interrupted.
READY
F,0 TO EXIT
Next injection or next vial inputs will no longer have any effect. Exit REMOTE CONTROL mode (F,0) to return
to READY status.
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21
DISPENSER FAILURE
PROGRAM VIALS
ERROR!(.............)
An erratic value has been entered; limits for this parameter are
shown in parenthesis on display. Enter correct value.
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22
APPENDIX 1 MAINTENANCE
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
17.
18.
19.
20.
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23
1.1
To keep rest volume small, the sample needle should stop close to the bottom of the sample vial. If
penetration depth needs adjustment, proceed as follows (see Figure A.1):
-
1.2
Refer to Appendix D.4 for adjustment of pressure range of RH 7010 injection valve.
1.3
FUSES
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24
1.4
1.5
1.6
INJECTION VALVE
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25
DESCRIPTION
The large volume sample tray is an option of the BASIC -MARATHON Autosampler. This option provides the
possibility to inject large volumes of sample with the DDS or by headspace pressure.
The option consists of:
1.8
INSTALLATION
Replace the standard installed tubing and needles by the tubing and needles of the LSV option. Follow
the procedure described in appendix A.
Check the needle penetration depth and if necessary adjust it, see appendix A.1.
Connect the drain to port 3, if the loop will be filled on time base by headspace pressure. It is also
possible to fill the loop with the DDS, but this will take less sample, but more time.
: 500 l
LSV
: TIME
The last SYSTEM SETTINGS parameter change is necessary for pressure controlled injections.
If the loop is filled with the DDS, be aware that a larger preflush volume (minimal 50 l) has to be
programmed to avoid cross contamination, because of the larger internal diameters of the tubing and
needle.
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26
LOOPVOLUME
:
SAMPLE VOLUME :
INJECTION MODE 1:
FLUSH
:
NEEDLE WASH
:
AUXILIARIES
:
INJ. MARKER PULSE :
RUN PARAMETERS
20 l
STD
FLUSHED
VOLUME
NO
NO
0.1 sec.
FIRST VIAL
LAST VIAL
NO. OF INJ/VIAL
PREFLUSH
ANALYSETIME
:
:
:
:
:
1
7
3
30 l
1:00 min
PUMP
FLOW
ELUENT
1.5 ml/min
distilled water
DETECTOR WAVELENGTH
254 nm
SAMPLE
Seven vials are placed in the BASIC -MARATHON tray, vial 1 and vials 4 till 7 are the vials with sample (50 ppm
Uracil), vials 2 and 3 are filled with eluent (blanks).
0060.103-14
27
Number
Peak Retention
Time
Peak
Area
====== ========= ==========
1
0.877
14.57611
2
2.181
14.65836
3
3.480
14.71600
4
13.363
14.59072
5
14.665
14.61937
6
15.964
14.62571
7
17.513
14.59504
8
18.815
14.65542
9
20.114
14.67614
10
21.680
14.58954
11
22.983
14.64998
12
24.281
14.49563
13
25.831
14.57137
14
27.129
14.58835
15
28.430
14.62109
Calculating the RSD% from the example integration results will give a RSD% of 0.36%.
0060.103-14
28
INTRODUCTION
The following standard sample loops are available; each loop is supplied with fittings for direct connection to
the valve:
Catalog number
mm
Tubing bore
inch
RH 7022
20
0.51
0.020
RH 7023
50
0.51
0.020
RH 7024
100
0.51
0.020
RH 7025
200
0.76
0.030
0060.103-14
29
Two stop pins and the rotor pin limit the rotation to 60 degrees. Four
conical spring washers and a thrust bearing are used to push the rotor
seal against the stator face with sufficient force to hold 7000 psi liquid
pressure. Three set screws in the stator are factory adjusted so that
when the three stator screws are fully tightened, the force between
stator and rotor is sufficient to hold 5000 psi. To increase maximum
operating pressure to 7000 psi, follow the directions in section D.3.
The isolation seal serves to prevent valve leakage from entering the spring and bearing region of the valve. It
does not serve any high-pressure sealing function. The maximum operating temperature of the Model RH
7010 is 150 C
To avoid clogging of the valve passages, it is recommended that filtered solvents be used and that sample
liquid be free of solids that might clog the loading tubes.
DISASSEMBLY
Disassembly of the valve for servicing of the liquid end is accomplished as follows:
a. Remove the three stator screws. Do not change the setting of the three set screws in the stator.
b. Refer to Figure D.2, and remove stator and stator ring from valve body by pulling axially to disengage
the various pins.
c. Isolation seal and bearing ring are usually left in place since they rarely need changing.
0060.103-14
30
1.12.2
ASSEMBLY
Reassembly of the valve requires putting all parts back together again as shown in Figure D.2, while
observing the following:
a. The rotor pin can be inserted in the rotor from one side only (the side without the peened closure
marks around the pin hole). Do not try to force the rotor pin in from the wrong side, and ensure that
the flat on the valve shaft faces away from the protruding end of the rotor pin.
b. Ensure that the rotor seal is correctly oriented as shown in Figure D.3, with rotor seal slots facing the
stator, and with the notch in the metal rim of the rotor seal in line with the rotor pin.
c. When replacing stator ring, be sure that the two stop pins are replaced in their holes in the stator ring;
then push the stator ring squarely onto the rotor assembly so that the stop pins enter the mating
holes in the body (with rotor pin between the two stop pins), and so that the isolation seal slips inside
the stator ring without hanging up.
d. Replace the stator by first pushing it onto the two pins on the stator ring and then adding the three
stator screws. Tighten each screw, a little at a time, to keep the stator surface parallel to the stator
ring surface. If the three set screws in the stator where not changed, then tighten the three stator
screws until all parts are firmly in their place. The three set screws will insure that the gap between
stator and stator ring is uniform and exactly as it was before disassembly.
e. If the set screws need adjusting because a new rotor seal was installed, or because leakage needed
to be stopped, ensure that each set screw is turned an equal amount so that after re-tightening the
stator screws, the gap between stator and stator ring is uniform all around. Refer also to Section D.3.
To prevent formation of salt crystals in the valve, flush out the flow passages and needle port with water
after using salt solutions.
1.13.2
The standard rotor seal is Vespel, a DuPont polyimide, which is exceptionally wear-resistant. It is, however,
susceptible to alkaline attack and deteriorates rapidly when used with solutions of pH 10 or higher. An
alternative material is available for alkaline applications (consult Rheodyne or local Spark dealer).
1.13.3
If valve passages are clogged, open them by removing the stator and cleaning the passages with a small
wire (maximum diameter 0.015 in.).
0060.103-14
31
0060.103-14
Tubing bore
(inches)
0.012
17 %
0.020
10 %
0.030
7%
32
Telephone
Fax
Local Dealer
Service Engineer
Address
Telephone
Fax
Comments:
0060.103-14
33
0060.103-14
34