BAS LC-4C User manual

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LC-4C
March, 2000
MF-9081 INSTRUCTION MANUAL
Electrochemical Detector
Bioanalytical
Systems, Inc
2701 Kent Avenue
West Lafayette
Indiana 47906
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MANUFACTURER’S NOTE
This instrument, either wholly or in part, is manufactured for research purposes only. Use for medical diagnosis is not intended,
implied or recommended by the manufacturer. Use for this purpose and accountability for the same rests entirely with the user.
The BAS logo is a registered trademark of Bioanalytical Systems, Inc.
Copyright March 2000
No portion of this manual may be reproduced without express, written permission of Bioanalytical Systems, Inc.
ALL RIGHTS RESERVED
Bioanalytical Systems, Inc.
2701 Kent Avenue
West Lafayette, IN USA 47906-1382
Phone: [phone]
Fax: [phone]
Never touch the electrochemical cell, the cell wires, or any part of the cell
while the detector is operating. Before touching the cell, make sure that
the cell switch on the front panel of the LC-4C amperometric controller is
in the “STANDBY” or “OFF” positions.Do not operate cell while dry, or
without connection to the reference electrode. Failure to obey these
warnings may result in damage to both the controller and the
electrochemical cell. Such damage is not covered by warranty.
DO NOT touch the cell unless the cell switch is in the “standby” position.
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Table of Contents
Section 1. Preface.................................................................................................................... 1
Section 2. Support Policy........................................................................................................3
2.1 User Updates.............................................................................................................3
2.2 Damaged Shipments.................................................................................................3
2.3 Product Warranty....................................................................................................... 3
2.4 Service Information...................................................................................................4
Section 3. Installation...............................................................................................................5
3.1 Inspection of Your Shipment.....................................................................................5
3.2 Power Requirements..................................................................................................9
3.3 Local Environment...................................................................................................10
3.4 Solvent Delivery Requirements................................................................................11
3.5 Arrangement of Components and Connections......................................................12
3.6 Chromatography Connections.................................................................................15
Section 4. Operating the Electrochemical Detector.................................................................23
4.1 LC-4C Front Panel..................................................................................................23
4.2 LC-4C Rear Panel....................................................................................................27
4.3 CC-5 Rear Panel......................................................................................................31
4.4 Routine Operating Procedure..................................................................................32
Section 5. Upgrading to a Dual Amperometric Detector.......................................................35
Section 6. Temperature-Stabilized Detector Operation .......................................................... 37
6.1 Rationale for Use.....................................................................................................37
6.2 Installation of Detector Preheater Module on CC-5................................................ 39
Appendix I. Electronic Troubleshooting.................................................................................41
Appendix II. Operation of the LC-4CE...................................................................................43
Index........................................................................................................................................I-1
Table of Contents
i
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Table of Contents
ii
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Section 1. Preface
This manual covers operation and service information for the LC-4C AMPEROMETRIC CON-
TROLLER. The LC-4C is a general purpose potentiostat which can be used for many pur-
poses. In this manual, we assume that a CC-5 cell compartment has also been purchased
with the LC-4C. There are other options that are described in BAS product literature and
companion manuals.
To use this detector, you will require a liquid chromatograph, a data recording device, and
appropriate end-fittings and tubing for the connection between column and detector. The LC-
4C is a component of the LC-44 detector and the modular BAS 480 series of liquid chroma-
tographs.
The information in this manual represents the combined experience of both the chemists
and engineers associated with LCEC from its inception. Background information can be
found in the “Principles of EC Detection and Troubleshooting Guide” (P/N MF-9083). Stand-
ard operating procedures are described in this manual; however, we have also sought to in-
clude the not-so-obvious experimental details which often make or break the user’s chances
of maintaining an extra edge of performance.
Section 1. Preface
1
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Several accessories may be added by the user or by service personnel at a later date:
1. The LC-4C Dual CELL LEADS (EW-8114) provide the additional cable to transform an ex-
isting single-electrode detector into true dual-electrode operation. A second LC-4C
controller (or an LC-3C/D/E Petit Ampère) must also be purchased. Installation instruc-
tions for dual-electrode operation are provided in Section 5.
2. A Preheater Module (EW-4055) may be added for better baseline stability at high detector
gain or in adverse environments. Its installation and use are described in Section 6.
3. An analytical Cartridge Column Heater (EF-1041) may also be added to the CC-5 compart-
ment. A removable panel on the back of the instrument is replaced by the LC-23C heat-
ing block, which holds up to 2 cartridges. This option is described in a separate man-
ual and product brochure. A variety of cartridge columns and guard columns are also
available from BAS.
4. A Rheodyne or UniJet microbore injection valve may be added by the user when the small
cover plate on the front of the CC-5 is removed.
5. ChromGraphsoftware with the DA-5 interface makes it possible to control the functions
of the LC-4C and also to implement techniques such as pulsed electrochemical detec-
tion.
The addition of options (3) and (4) to your electrochemical detector will convert it into the
basis of a complete liquid chromatograph.
Section 1. Preface
2
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Section 2. Support Policy
2.1 User Updates To activate your warranty and receive product update information news and valuable infor-
mation related to this and other BAS products, fill out and return the Warranty Enrollment
Card which was shipped with the instrument. We would like to know who you are, and what
more you want to know about electrochemical detection.
2.2 Damaged Shipments Breakage of any part of this instrument during shipping should be reported immediately to
BAS Customer Service. You must retain the original packing box and contents for inspec-
tion by the freight handler. BAS will replace any new instrument damaged in shipping with
an identical product as soon as possible after the claim filing date. Claims not filed within
30 days after the shipping date will be invalid.
Do not return damaged goods to BAS without first contacting Customer Service for a Return
Authorization Number (RA#). When a defective part is returned to BAS, the RA# immediately
identifies you as the sender and describes the item being returned. Bioanalytical Systems re-
fuses all unauthorized return shipments.
2.3 Product Warranty BAS products are fully warranted against defects in material and workmanship. The LC-4C
amperometric controller is unconditionally warranted for one year from date of shipment,
except when failure is due to obvious abuse or neglect, unauthorized tampering, procedures
not described in manuals, or improper connection of electronic units to other components.
Electrochemical cells are warranted for 60 days from date of shipment under the same exclu-
sions. Chromatographic columns and injection valves are warranted for 30 days. The follow-
ing items are not covered under any warranty: carbon paste, activated aluminum oxide,
lamps, panel lights, fuses, pump seals, valve seals, and reference electrodes.
For any product expressly covered under this warranty, BAS is liable only to the extent of re-
placement of defective items. Bioanalytical Systems, Inc. shall not be liable for any personal
injury, property damage, or consequential damages of any kind whatsoever. The foregoing
warranty is in lieu of all other warranties of merchantability and fitness for a particular pur-
pose.
Section 2. Support Policy
3
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2.4 Service Information Bioanalytical Systems provides a skilled service staff available to solve your technical prob-
lems if an equipment-oriented problem should arise. For further details, call customer serv-
ice personnel (1-800-845-4246), who will route your problem to the correct individual. Fol-
lowing discussion of your specific difficulties, an appropriate course of action will be de-
scribed and the problem resolved accordingly.
DO NOT RETURN ANY PRODUCTS FOR SERVICE UNTIL A RETURN AUTHORIZATION
NUMBER (RA#) HAS BEEN OBTAINED. The RA# identifies you as the sender and describes
the problem you are having in full detail. Turnaround time on service can be quoted to you
at the time your RA# is issued, although we can not determine the actual amount of service
required until we have received your unit and diagnosed the problem. All correspondence
and shipments should be sent to:
RA # , Service Department
Bioanalytical Systems, Inc.
2701 Kent Avenue
West Lafayette, IN 47906
Section 2. Support Policy
4
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Section 3. Installation
3.1 Inspection of Your
Shipment After carefully unpacking the instrument, check the contents of the packages and inspect for
breakage (see Section 2). Electrodes and flow cell components (if purchased) are packaged
in a gray box. Assembly of these various parts will be outlined in the following chapters.
Another gray box, containing the PK-4 polishing kit, is provided with the flow cell. The pol-
ishing kit includes an Electrode Manual, which describes electrode polishing, use of gas-
kets, and reference electrode assembly.
Please retain the shipping box and packing material until you have fully tested the unit to
be certain that no damage was incurred during shipping.
If there are any discrepancies, retain the packing slip and contact BAS Customer Service for
assistance (see Section 2).
Figures 3.1–3.4 show the various components of the complete system.
Figure 3.1. Top view of CC-5 flowcell configured as part of a BAS 480 system. Refer to Table 3.1 for descriptions.
PREHEATER POWER
HIGHLOW 10
5
4
21
22
19 78
14 13
9
28
23
24 20
18
25
3
27
16
30
11
15
29
26
31
Section 3. Installation
5
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Table 3.1 Parts of the assembled LC-44 Amperometric Detector (refer to Figures 3.1–3.4).
Ref. # Description
1 LC-4C Amperometric Detector
2 CC-5 Flowcell Compartment
3 Injection Valve (installed only in the BAS 480 liquid chromatographs, but available from BAS for installation by customers
who wish to upgrade their detectors and use them as the basis for an LC system)
4 Utility Panel (this is removed when the unit is upgraded for cartridge column temperature control, model LC-22C/23C)
5 Optional Preheater Module (120 V/60 Hz or 240 V/50 Hz)
6 Optional LC-22C Temperature Controller for Detector Preheater Module or Cartridge Column Heater Temperature Control
(120 V/60 Hz or 240 V/50 Hz)
7 Stainless Steel Auxiliary Electrode
8 Reference Electrode Retainer (packaged in detector accessories box)
9 Working Electrode Clamp and backing plate (packaged in detector accessories box)
10 Single Electrode Cable (dual electrode cable has two working electrode leads)
11 Generator Electrode Lead
12 Power Cord/Fuse location on LC-4C Controller
13 Dual Working Electrode Block (glassy carbon is standard)
14 Reference Electrode (pkg. of 3 RE-4 Ag/AgCl/gel reference electrodes is provided in detector accessories box)
15 TG-2M Electrode Gasket (4 of these translucent gaskets are in the same small box with the electrode jumper and foam
sheets)
16 Dual Electrode Jumper (use is optional) (packaged with the electrode gaskets)
17 PTFE Inlet Tubing, 0.25-mm i.d. (not shown; packaged with detector accessories) (not used with cell preheater)
18 20-µL Loop (not provided with detector purchases, BAS 480 ONLY)
19 PEEK Fingertight Nut (cell outlet)
20 Valve to column inlet tube (already prepared with correct end fittings, not provided with detector purchases,
BAS 480 ONLY)
21 PEEK Fingertight Nut for column to cell inlet tubing
22 Inlet Tubing, Stainless Steel (for preheater) or Teflon (without preheater, #17)
23 Analytical Column, 100 ×3.0 mm, 3 µm ODS (BAS P/N MF-8954) (not provided if only detector is purchased)
24 Column End Fittings (not provided with detector purchases, BAS 480 ONLY)
25 Connecting Tube from PM-80 Pump to Injection Valve (when installed)
26 PTFE Outlet Tubing (0.5-mm i.d.) (note the small piece of larger tubing located midway on the tube, used to secure the
tubing in a notch on the right side of the cell compartment; unless the tubing is secured in the notch,
it may be pinched closed when top is lowered; see #30)
27 Notch in cabinet for tubing connection from pump
28 Notch in cabinet to allow use of longer columns (e.g., conventional 25 cm columns) which will extend outside the CC-5
cabinet
29 Notch in cabinet for cell outlet tube (be sure to press the portion of tube with a thicker, Tygon sheath into this slot)
30 Output cable to chart recorder
31 Optional ground connector
Section 3. Installation
6
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Figure 3.2. Front view of LC-44 Detector with temperature control. Refer to Table 3.1 for descriptions of numbered components.
Figure 3.3. Rear view of LC-44 Detector with temperature control. Refer to Table 3.1 for descriptions of numbered compo-
nents.
SETPOINT
HEATER 1
SETPOINT
HEATER 2
DISPLAY
SELECT
DISPLAY
UP DN
POWER
FILTER
POTENTIAL MANUAL OFFSET
+_
RANGE
+_
OFF +
_
+
_100
10
1000
ON
ZERO
OFF
ON
TEST
STBY
AUTO ZERO MARKER
CELL
6
1
2
LC-44 SERIES
BAS 480 ANALYZER
MADE IN USA
CAUTION:
TO PREVENT ELECTRICAL SHOCK, DO NOT REMOVE
COVER. NO USER SERVICEABLE PARTS INSIDE.
REFER SERVICING TO FACTORY PERSONNEL.
CONNECT ONLY TO
"LC-23C PROBE" ON
BAS LC-22C
CONNECT ONLY TO "HEATER
OUTLET" ON BAS LC-22C
6
1
2
1V GND 10mV
OXD
RDN
REMOTE
CELL
REZERO MARK 1V 10mV EIN GND EAPP GND IGEN GND WGEN GND GND GND GND
EGEN
ADJ
A
I
BCDEFGH
JKLMNOP
LC-4C
Made in USA
REMOTE OUTPUT APP E GEN
GND
LC-22C
Made in USA
GND
REMOTE
ADDRESS OUTPUT
PROBE 1 PROBE 2
HTR 2HTR 1 F1
F2
FUSE
PULL 12
Section 3. Installation
7
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Figure 3.4. BAS 480 Liquid Chromatograph with PM-80 Pump at bottom level. The column and injection valve are mounted in
the detector compartment at the top level. Refer to Table 3.1 for descriptions of numbered components.
SETPOINT
HEATER 1 SETPOINT HEATER 2
DISPLAY
SELECT
DISPLAY
UP DN
POWER
FILTER
POTENTIAL MANUAL OFFSET
+_
RANGE
+_
OFF
+
_
+
_100
10
1000 ON
ZERO
OFF
ON
TEST
STBY
AUTO ZERO MARKER
CELL
POWER
6
1
23
40
30
20
10
0
OFF
40
30
20
10
0
MIN MAX
PRESSURE (PSI)
FLOW RATE
REMOTE
LOCAL
0-0.5 mL/MIN
0-5 mL/MIN
STOP
START
Section 3. Installation
8
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3.2 Power Requirements The LC-4C and (optional) LC-22C instruments are operated with either 100, 120, 220, or
240 V AC and 50 or 60 Hz power, but the correct voltage must be selected at the cord con-
nector before use. Maximum power consumption is less than 120 watts. The units should
be shipped with the voltage option already matched to the destination, but double check
the settings to be sure they match the power requirements of your laboratory.
Figure 3.5. Power selection on the LC-4C.
Should the power option need to be changed, unplug the line cord and slide the plastic
window to the left (as shown in Figure 3.5). The orientation of the small circuit board now
exposed in this socket determines the voltage option. If the voltage labeled on the outer
edge of this board is incorrect, pull out the board and turn it (either by rotation or inver-
sion) so that the desired voltage is readable. Reinsert the board and push the fuse holder
back into the cavity. Also check Table 3.2 to see that the fuse is the proper rating.
Table 3.2. Correct fuse selection.
Voltage Fuse
110 V/60 Hz 1 A/SB
100 V/50 Hz 1 A/SB
220–240 V/50 Hz 500 mA/SB
FUSE
PULL
FUSE
VOLTAGE SELECT PCB
Section 3. Installation
9
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3.3 Local Environment Amperometric detection is a highly sensitive technique. The currents measured typically fall
in the picoampere or nanoampere range. Hence, smooth operation can easily be influenced
by electrical disturbances in the environment. Also, since detection is due to a chemical re-
action, its response (and baseline drift) is temperature-dependent; this is very noticeable at
high detector gain.
The new design of the LC-44 Amperometric Detector decreases the influence of these prob-
lems. It offers significant advantages over earlier models. The Preheater Module, when con-
trolled by the LC-22C Temperature Controller, eliminates long-term drift due to room tem-
perature fluctuations. The new flowcell design is well shielded from stray capacitance prob-
lems; moreover, it is wholly contained within an easily opened, grounded compartment.
When selecting a location for the detector, follow these guidelines:
1. Connect the entire chromatograph (pumps, detector, recorder, temperature controller, etc.)
to the same grounded power line. This ensures adequate ground to all components of
the system and eliminates the possibility of a ground loop, which could produce base-
line noise. Also, select a power line that is lightly used. Other laboratory instruments
such as ovens, vortex mixers, centrifuges, and large motors may cause spikes in the
power line.
2. Locate the chromatograph on a stable bench. Vibrations can hamper the performance of
any sensitive instrument.
3. Select a room where the temperature remains stable throughout the day. Avoid installing
the chromatograph near windows, air ducts, ovens, or refrigerators.
4. Place the chromatograph away from busy, congested areas. Remote, isolated areas are
best for high-sensitivity work.
5. Avoid very dry areas and areas that are carpeted. Static electricity can affect instrument
performance. Antistatic floormats and benchmats are useful if spiking caused by static
charge is a problem.
6. Avoid areas where radio frequency interference is likely. Beeper-type paging devices can
be a problem in some installations.
Section 3. Installation
10
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3.4 Solvent Delivery
Requirements Electrochemical detection requires a solvent delivery system that delivers a constant rate of
flow. The detector will respond to long-term and short-term changes in flow rate. A gradual
increase in flow rate will produce a gradual increase in baseline current response. Likewise,
a rapid cycling in flow rate will cause a sinusoidal baseline response. Both types of flow
rate changes will severely limit detector performance. Ideally, this instrument will be part of
a BAS 480 Liquid Chromatograph. The BAS PM-80 Pump in this system provides excellent
flow-rate precision and minimal pump pulsation. The PM-80 Pump may be connected di-
rectly to the injection valve port (see #26 in Figure 3.1).
The PM-80 pump includes a pulse damper to reduce short-term flow rate fluctuations. For
other pumps, however, an external pulse damper may improve performance. BAS offers a
rugged pulse damper (P/N MF-4000) that is suitable for use with any LCEC system (see
Figure 3.6). It is connected between the pump and injection valve. The damper contains a
flattened coil of 1/4" tubing which expands at high pressures and flattens at low pressures.
This effectively steadies the flow rate over time.
Figure 3.6. MF-4000 Pulse Damper.
Section 3. Installation
11
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3.5 Arrangement of
Components and
Connections
NOTE: Should you be installing as retrofits either the LC-4C Dual Upgrade Kit or the CC-5
Preheater Module, refer to the installation instructions in Section 5 or Section 6.2, respec-
tively. The instructions below are for startup in a new installation.
1. Arrange the various components per Figure 3.7. The LC-4C controller(s) must always be
placed immediately under the CC-5 compartment. The CC-5 has been sealed along all
joints. If an accidental leak should occur, the compartment could hold over 500 mL of
liquid before overflowing to the components below.
2. If you purchased the CC-5 compartment with Preheater Module, connect the two cables
on the preheater to the LC-22C Temperature Controller (see Figure 3.8). The large
power cable from the Preheater Module plugs into either of the two matching connec-
tors on the LC-22C rear panel. The control lead with color coded banana plugs con-
nects to the corresponding set of banana jacks. If you plugged the power cable into
the HEATER1 connector, plug the banana plugs into the PROBE1 jacks. If you used
HEATER2 for the power cord, connect the banana plugs to the PROBE2 jacks.
3. Connect the electrode cable (single or dual) from the CC-5 to the LC-4C(s) per
Figure 3.9.
4. If you will be using the generator electrode, connect the insulated lead of the GEN detec-
tor cable to the WGEN connector on the 16-position terminal strip of the LC-4C back
panel. Connect the noninsulated lead to the GND connector. If you will be monitoring
the output of the GEN electrode, connect leads to IGEN and GND. (CAUTION: Refer to
page 28 under GEN for more details.)
5. For additional support, two brackets and four screws are provided for every additional
component in the “stack” of instruments. Brackets are attached at the back panels in
the positions shown in Figure 3.9.
Figure 3.7. Schematic for stand-alone electrochemical detectors and BAS 480 components.
Section 3. Installation
12
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Figure 3.8. Connection of cables from CC-5 Preheater Module to LC-22C Temperature Controller.
Figure 3.9. Connection of leads to LC-4C(s) for single (left) and dual (right) operation.
LC-44 SERIES
BAS 480 ANALYZER
MADE IN USA
CAUTION:
TO PREVENT ELECTRICAL SHOCK, DO NOT REMOVE
COVER. NO USER SERVICEABLE PARTS INSIDE.
REFER SERVICING TO FACTORY PERSONNEL.
CONNECT ONLY TO
"LC-23C PROBE" ON
BAS LC-22C
CONNECT ONLY TO "HEATER
OUTLET" ON BAS LC-22C
1V GND 10mV
OXD
RDN
REMOTE
CELL
REZERO MARK 1V 10mV EIN GND EAPP GND IGEN GND WGEN GND GND GND GND
EGEN
ADJ
A
I
BCDEFGH
JKLMNOP
LC-4C
Made in USA
REMOTE OUTPUT APP E GEN
GND
LC-22C
Made in USA
GND
REMOTE
ADDRESS OUTPUT
PROBE 1 PROBE 2
HTR 2HTR 1 F1
F2
FUSE
PULL
LC-44 SERIES
BAS 480 ANALYZER
MADE IN USA
CAUTION:
TO PREVENT ELECTRICAL SHOCK, DO NOT REMOVE
COVER.NO USER SERVICEABLE PARTS INSIDE.
REFER SERVICING TO FACTORY PERSONNEL.
CONNECT ONLY TO
"LC-23C PROBE" ON
BAS LC-22C
CONNECT ONLY TO "HEATER
OUTLET" ON BAS LC-22C
1V GND 10mV
OXD
RDN
REMOTE
CELL
REZERO
MARK
1V
10mV
EIN
GND
EAPP
GND
IGEN
GND
WGEN
GND
GND
GND
GND
A
I
BCDEFGH
JKLMNOP
REMOTE OUTPUT APP E GEN
LC-44 SERIES
BAS 480 ANALYZER
MADE IN USA
CAUTION:
TO PREVENT ELECTRICAL SHOCK, DO NOT REMOVE
COVER.NO USER SERVICEABLE PARTS INSIDE.
REFER SERVICING TO FACTORY PERSONNEL.
CONNECT ONLY TO
"LC-23C PROBE" ON
BAS LC-22C
CONNECT ONLY TO "HEATER
OUTLET" ON BAS LC-22C
1V GND 10mV
OXD
RDN
REMOTE
CELL
REZERO
MARK
1V
10mV
EIN
GND
EAPP
GND
IGEN
GND
WGEN
GND
GND
GND
GND
A
I
BCDEFGH
JKLMNOP
REMOTE OUTPUT APP E GEN
1V GND 10mV
OXD
RDN
REMOTE
CELL
REZERO
MARK
1V
10mV
EIN
GND
EAPP
GND
IGEN
GND
WGEN
GND
GND
GND
GND
A
I
BCDEFGH
JKLMNOP
REMOTE OUTPUT APP E GEN
Section 3. Installation
13
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6. Connect the output leads to the strip-chart recorder, integrator, or data station. See Sec-
tion 4.2 for more information on the LC-4C’s rear panel connections.
a. On each LC-4C, there are four sets of output jacks (see Figure 3.9). Use the
green and black sockets for a 1.0 V recording device. Use the blue and
black sockets for a 10 mV recording device. Alternatively, use the bare-wire
connectors at the OUTPUT section of the 16-position terminal strip on the
LC-4C back panel.
b. Flip the OXD/RDN switch on the rear panel to the mode you are using
(oxidation or reduction). If you are unfamiliar with the meaning of these
terms, you will find more information in BAS’ “Principles of EC Detection
and Troubleshooting Guide” (P/N MF-9083). Correct positioning of this
switch ensures a positive output signal. This switch can be used to reverse
the direction (polarity, see Section 4.2, page 27) of the pen travel (output
signal) on the chart recorder.
c. At the recorder, connect the leads as follows:
White lead (red plug): “+” or “HI” recorder input
Black lead (black plug): “–” or “LO” recorder input
Black jumper (green plug): recorder ground terminal
(LO is thus jumpered to GND)
GROUNDING NOTE: It is important that the third plug on the recorder leads be
connected to ground. If your data system does not have a ground, connect
a wire (included: part #32, Figure 3.1) from the OUTPUT GND of the
terminal strip to the threaded hole at the upper left corner of the rear panel.
If this is a dual-detector system, also connect a wire between the OUTPUT
GNDs of the two LC-4Cs.
d. Using the ZERO ADJUST or ZERO CHECK on your recorder, zero the recorder
at the appropriate side of the chart paper.
NOTE: If you have an integrator or data station with wide dynamic range, match the output
of the LC-4C to the input range of your data device. The LC-4C has both 1 V and 10 mV
outputs, making it compatible with the majority of popular chart recorders, integrators and
workstations. There is a built-in overange on each output that is 10 times the nominal
value (e.g., the range on the 10 mV output is from 0 to 100 mV). Use the RANGE switch on
the LC-4C Detector to keep the signal you record within the range of your recording device.
7. The entire LC system should be connected to the same ground point. This point should
be the safety ground (third wire) of the power circuit you are using.
To ensure a properly grounded system, have a qualified electrician measure the resis-
tance between each component chassis and circuit ground. The resistance should be
less than 1 Ω. If it is not, ground the component chassis together with a jumper wire.
Section 3. Installation
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3.6 Chromatography
Connections 1. Place the interconnected detector components next to the column outlet from the chroma-
tograph, or install your column inside the CC-5 (Figure 3.1). Several slots in the CC-
5 chassis along the sides and back provide tubing access, no matter what the orienta-
tion. On a BAS 480 system, the detector components may sit directly on top of the
PM-80 Pump. See Figures 3.2–3.4 and 3.7 for ideas.
MICROBORE INSTALLATION: A microbore column should be connected directly
between the injection valve and the flow cell. You must use a special UniJetinjec-
tion valve (MF-4161) that uses nonconducting fittings. Connect the microbore column
to the correct output port of this valve with a high-pressure PEEK fingertight fitting (MF-
4165). Release the flow cell from the CC-5 cabinet by undoing the hex screw. Slide
the flow cell toward the column until the column bottoms in the entry port, and secure
with another fingertight fitting.
NOTE: It will be necessary to thoroughly degas the mobile phase prior to use. This is
particularly important when the cell preheater or column heater is set at 35 °Cor
higher. The warmth reduces dissolved gas solubility to the point where bubbles may
appear in the tubing to and from the electrochemical cell.
If you are using a flow restrictor to provide back pressure, be sure not to exceed ap-
proximately 100 psi (6 atm).
2. Flush the system with 200 mL of acetonitrile/water (40:60, v:v) and divert all column ef-
fluent to a waste receptacle. This flushing step cleans any chemicals remaining from
previous injections from the column, and also “wets” the hydrophobic stationary
phase for reproducible separations. For new columns see below.
NOTE: Always connect a BAS column so fluid flows in the direction you read the label
(because the inlet and outlet frits have different pore sizes).
Most new BAS columns must be washed before use.* Washing will not only remove
residual packing solvents, but will give sufficient time for elimination of bubbles from
the system. For C18 and C8stationary phases, this step also “wets” the stationary
phase surface. To wash the column, pump at least 200 mL neat acetonitrile through it.
Then pump 50 mL 40:60 acetonitrile:water through it to remove the neat acetonitrile.
(It is important not to mix neat solvents and mobile phase in the system or column,
since buffer salts may precipitate.) Then pump mobile phase.
The column should be thoroughly equilibrated with mobile phase before connecting
the detector. If you are using an ION-PAIRING REAGENT in your mobile phase (e.g.
Sodium Octyl Sulfate), you should allow up to 8 hours for equilibration at a flow rate
of 1 mL/min.
* Exceptions include enzyme columns, acetylcholine columns, and preloaded catecho-
lamine columns, which must never be washed with solvents. SepStik microbore col-
umns should be washed with acetonitrile for 15 minutes at microbore flow rates.
Section 3. Installation
15
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