Bio-Rad BioLogic QuadTe Detector and Components User Manual
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SCAN
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BioLogic
uadTec™
Q
UV/VIS Detector
Instruction Manual
SAFETY
Caution/Warning
Disconnect the QuadTec’s power cable from its power source before servicing. Refer to
Chapter 5, Maintenance, for instructions on servicing the BioLogic QuadTec system.
The BioLogic QuadTec UV/Vis detector is certified to meet the I.E.C. 1010-1* standard for safety of
laboratory equipment. Certified products are safe to use when operated in accordance with the instruction
manual. This safety certification does not extend to other chromatography equipment or accessories not
I.E.C. 1010 certified, even when connected to this QuadTec UV/Vis detector.
The QuadTec UV/Vis detector conforms to the class A standards for Electromagnetic Emissions, intended
for laboratory equipment applications. It is possible that emissions from this product may interfere with some
sensitive appliances when placed near or on the same circuit as those appliances. The user should be
aware of this potential and take appropriate measures to avoid interference.
This instrument should not be modified or altered in any way. Alteration of this instrument will void the
manufacturer’s warranty, void the I.E.C. 1010 certification, and create a potential safety hazard for the user.
Bio-Rad is not responsible for any injury or damage caused by the use of this instrument for purposes other
than for which it is intended, or by modifications of the instrument not performed by Bio-Rad or an authorized
agent.
*I.E.C. 1010 is an internationally accepted electrical safety standard for laboratory instruments.
SAFETY
!
TABLE OF CONTENTS
TABLE OF CONTENTS
Safety
Chapter 1 General Description .................................................................................................. 1-1
1.1 Using this Manual........................................................................................................ 1-2
1.2 Optical Path of the QuadTec UV/Vis Multi-wavelength Detector ................................ 1-2
1.3 Unpacking ................................................................................................................... 1-3
1.4 Preparing the QuadTec Detector for Operation .......................................................... 1-4
1.4.1 Installation of the Flow Cell ............................................................................ 1-4
1.4.2 Power Up and Self-Test.................................................................................. 1-5
Chapter 2 Using the QuadTec with the BioLogic DuoFlow™ System................................... 2-1
2.1 Instrument Control Module (ICM)................................................................................ 2-2
2.2 Electrical Connections................................................................................................. 2-3
2.3 Plumbing Connections ................................................................................................ 2-4
2.4 System Power-up ........................................................................................................ 2-5
2.5 System Operation ....................................................................................................... 2-5
2.5.1 Manual Screen ............................................................................................... 2-5
2.5.2 Setup Editor Screen ....................................................................................... 2-8
2.5.3 Run Screen and Chromatogram Settings ...................................................... 2-8
Chapter 3 Using the QuadTec with the BioLogic HR System................................................ 3-1
3.1 Electrical Connections ............................................................................................... 3-2
3.2 Plumbing Connections ................................................................................................ 3-3
3.3 System Operation ...................................................................................................... 3-3
3.3.1 Using the QuadTec Faceplate Keys............................................................... 3-3
3.3.2 Changing Operating Parameters.................................................................... 3-4
3.4 Selecting Wavelengths at the QuadTec ...................................................................... 3-4
3.5 Configuring the Biologic HR Software......................................................................... 3-4
3.6 Writing a Method ........................................................................................................ 3-5
3.7 QuadTec Detector Setup Menu................................................................................... 3-5
Chapter 4 Stand-Alone Mode..................................................................................................... 4-1
4.1 System Operation ....................................................................................................... 4-1
4.1.1 Using the QuadTec Faceplate Keys............................................................... 4-1
4.1.2 Changing Operating Parameters.................................................................... 4-1
4.1.3 Accessing Menus ........................................................................................... 4-1
4.2 Display and Menu Structure........................................................................................ 4-2
4.2.1 The QuadTec Detector Setup Menu .............................................................. 4-2
4.2.2 LINK Menu ..................................................................................................... 4-5
4.2.3 VIEW Menu .................................................................................................... 4-6
4.3 Stand-Alone Control of the QuadTec .......................................................................... 4-6
4.3.1 Selecting Wavelength(s)................................................................................. 4-7
4.3.2 Output Options ............................................................................................... 4-7
4.3.3 Events ............................................................................................................ 4-7
4.4 Programming the QuadTec UV/Vis Detector .............................................................. 4-8
4.4.1 Creating Programs ......................................................................................... 4-8
4.4.2 Deleting Programs.......................................................................................... 4-8
4.4.3 Time Programming of Absorption Wavelengths............................................. 4-9
4.4.4 Programming Events for Control of a Fraction Collector................................ 4-10
4.4.5 Program Execution......................................................................................... 4-11
4.4.6 Program Links ................................................................................................ 4-11
i
4.5 Scanning UV Spectra.................................................................................................. 4-12
4.5.1 Scan Menu ..................................................................................................... 4-12
4.5.2 Scan Auto-Zero .............................................................................................. 4-13
4.5.3 Performing Wavelength Scans ....................................................................... 4-13
4.5.4 Output of Scan Data ...................................................................................... 4-13
4.6 QuadTec Detector Wake-up Program ......................................................................... 4-14
4.7 GLP Menu ................................................................................................................... 4-14
4.8 Connecting Other Instruments to the QuadTec Detector ........................................... 4-15
4.8.1 Using the Event and Remote Connectors...................................................... 4-15
4.8.2 RS-232 Serial Interfaces................................................................................ 4-15
Chapter 5 Maintenance............................................................................................................... 5-1
5.1 Checking the Status of the Deuterium Lamp.............................................................. 5-1
5.2 Changing the Lamp..................................................................................................... 5-2
5.3 Cleaning the Flow Cell ................................................................................................ 5-4
Chapter 6 Troubleshooting the QuadTec with the BioLogic DuoFlow System.................... 6-1
Appendix A Specifications............................................................................................................ A-1
Appendix B Wavelength Selection for Typical Biological Molecules........................................ B-1
Appendix C Ordering Information ................................................................................................ C-1
TABLE OF CONTENTS
ii
1.0 GENERAL DESCRIPTION
The BioLogic QuadTec multi-wavelength UV/Vis detector adds versatility to your biologic chromatography
system. The QuadTec detector has a wavelength range of 190–370 nm using the standard deuterium lamp
and 370–740 nm with the optional halogen lamp. The standard deuterium lamp is required for the detection
of peptides, proteins, DNA and other nucleotides.
Wavelengths are selected through the use of a moveable grating monochromator with an accuracy of ±1 nm
in 1 nm steps. The bandwidth is 8 nm or better. When using the deuterium lamp for the monitoring of
wavelengths >380 nm, an automatic cutoff filter is activated. This filter suppresses secondary grating orders
at wavelengths > 380 nm and thus ensures the selectivity of the measurement. Because very little light of
wavelengths greater than 400 nm is emitted from a deuterium lamp, the halogen lamp is required for routine
detection above 380 nm.
Figure 1-1. BioLogic QuadTec UV/Vis Detector
When connected to the BioLogic DuoFlow system via an Instrument Control Module (ICM), four wavelengths
may be monitored simultaneously and displayed on the Biologic DuoFlow Controller. In this configuration, the
QuadTec is controlled from the DuoFlow Controller.
When connected to the Biologic HR system via two Signal Import Modules (SIM-HR), two wavelengths may
be monitored and displayed simultaneously. In this configuration, wavelength selection and zero baseline
functions must be performed from the QuadTec UV/Vis Detector faceplate.
In stand-alone mode, two simultaneous wavelengths may be output to a chart recorder. The SCAN function
provides you with spectral information of a sample over the entire wavelength range and data may be output
to a chart recorder. This feature is NOT supported by the BioLogic system software.
INTRODUCTIONSYSTEM OVERVIEW
1-1
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Time
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Run 1
1 2 3 4 5
Signal 1
[ au ]
1
(nm)
Signal 2
[ au ]
2
(nm)
Setup
GLP
SCAN
6 7 8 9 10
1.1 USING THIS MANUAL
The instructions provided in this manual are for use of the QuadTec Detector with the following systems:
• The BioLogic DuoFlow chromatography system
• The BioLogic HR chromatography system
• Stand-alone use
1.2 OPTICAL PATH OF THE QUADTEC UV/VIS MULTI-WAVELENGTH DETECTOR
The light emitted from the lamp [1] is focussed by a lens [2] and a slit [4]. If a deuterium lamp is used and
wavelengths >380 nm are chosen, an automatic edge filter [3] is employed. The light passes to a grating
monochromator [6] for selection of the desired wavelength. The beam is then split by a semi-transparent
mirror [7]. Part of the beam passes to the photo diode reference signal [8] and optical absorption is
measured. The other part of the beam passes through the flowcell and sample and the light intensity is
measured at the sample photo diode [10]. The difference or the ratio of the two signals is calculated to
provide information about sample components.
Figure 1-2. Theory of Operations
INTRODUCTION SYSTEM OVERVIEW
1-2
1. LAMP (DEUTERIUM OR HALOGEN)
2. LENS SYSTEM (QUARTZ OPTICS)
3. EDGE FILTER
5. CONCAVE MIRROR
10. SAMPLE PHOTO DIODE
4. SLIT
6. MOVABLE GRATING
MONOCHROMATOR
7. SEMI-TRANSPARENT MIRROR
8. REFERENCE PHOTO DIODE
9. FLOW CELL
1.3 UNPACKING
When you receive the QuadTec detector and system accessory kit, carefully inspect the shipping containers
for any damage, which may have occurred in shipping. Severe damage to a container may indicate damage
to its contents. If you suspect damage to the contents, immediately file a claim with the carrier in accordance
with their instructions. If necessary, contact your local Bio-Rad office or Bio-Rad technical support at
1-800-4-BIORAD.
• QuadTec Detector for Use with the BioLogic DuoFlow System
Includes a 3 mm PEEK flow cell, system cable 25 (RS232), 10-32 fingertight fittings (quantity 4).
See Figure 1-3. Also includes an instrument control module (ICM), system cable 26 (ICM power
cable), system cable 17 (BioLogic communication cable), DuoFlow software version 3.0, BioLogic
DuoFlow Instruction Manual chapter inserts, QuadTec instruction manual, power cord.
• BioLogic HR Accessory Kit for Use with the QuadTec Detector
Includes system cable 17 (BioLogic communication cable), SIM-HR (quantity 2), system cable 24
(QuadTec analog out cables to SIM-HR; quantity 2), QuadTec instruction manual, power cord.
• QuadTec Detector for Stand-Alone Use
Includes a 3 mm PEEK flow cell, system cable 25 (RS232), 10-32 fingertight fittings (quantity 4),
QuadTec instruction manual and power cord.
Figure 1-3. Package Contents of the QuadTec Detector
INTRODUCTIONSYSTEM OVERVIEW
1-3
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3
0
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AUTO
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SCAN
1.4 PREPARING THE QUADTEC DETECTOR FOR OPERATION
1.4.1 Installation of the Flow Cell
The QuadTec detector is shipped with a “dummy” flow cell installed. Before operating the detector, you need
to install the biocompatible 3 mm PEEK flow cell. Refer to Figure 1-4 and the procedure below.
1. Make sure the detector power is OFF.
2. Loosen the two knurled screws on the front of the flow cell and gently pull them out. This allows the
flow cell housing to slide out.
3. Remove the dummy cell by gently pulling it upward. Save the dummy cell in a secure place. It is
required for measuring signal and reference output values for the lamps.
4. Insert the new flow cell and make sure that the engraved specifications point towards the user and
that the fixing hole on the back side of the cell meets the corresponding metal pin of the detector’s
housing.
5. Slide the complete system towards the detector, insert the two knurled screws, and tighten them by
hand.
6. Attach the 10-32 fingertight fittings and tubing to the inlet and outlet ports and use a syringe to rinse
the flow cell with 10–20 ml of 100% analytical grade methanol, followed by 5 ml DI water.
Figure 1-4. Installing the Flow Cell
INTRODUCTION SYSTEM OVERVIEW
1-4
INLET
SCAN
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Events: 2
off 0 on /
Time
(min)
Signal 1
[ au ]
Signal 2
[ au ]
1
(nm)
Setup
GLP
2
(nm)
FLOW CELL
3
0
OUTLET
FLOW CELL HOUSING
1.4.2 Power Up and Self-Test
The QuadTec detector is equipped with a universal power supply, which operates with supply voltages from
90 to 260 Volts AC. A manual setting of the supply voltage is not required.
Caution! Make sure to use a properly grounded power outlet and the power cable provided with the
system.
Before connecting the QuadTec to the Biologic DuoFlow or BioLogic HR system, perform the following
operation:
1. Connect the power cable to the QuadTec and the grounded power outlet, and then switch on the
device using the ON/OFF switch on the rear of the unit.
2. The display should show: Kernel v1.0, Bio-Rad BioLogic QuadTec UV-Vis 1.50.
Note: The firmware revision number may change.
3. Upon power-up, the QuadTec will run through a self-test which includes a check of the electronic
functions. Successful completion of self-test is indicated by SYSTEM TEST OK.
4. The QuadTec then runs through a wavelength calibration routine. A noise will be heard as the
monochromator moves. If the calibration is successful, the main menu will appear (see page 4-2
Figure 4-1).
If the detector calibration fails or takes an excessively long time, may be a sign that the lamp needs
replacing. See Chapter 6, Troubleshooting.
If the QuadTec self-test is successful, turn off the instrument, and then connect it to your chromatography
system using the instructions given on the following pages.
INTRODUCTIONSYSTEM OVERVIEW
1-5
2.0 USING THE QUADTEC WITH THE BIOLOGIC DUOFLOW SYSTEM
The QuadTec UV/Vis detector provides a powerful addition to the analytical capabilities of the BioLogic
DuoFlow system. An instrument communication module (ICM) provided with the detector permits complete
programming and control by the BioLogic DuoFlow software version 3.0 or greater.
When the QuadTec is attached to the BioLogic DuoFlow system via the ICM, up to four wavelengths can
simultaneously be monitored and displayed. The BioLogic DuoFlow displays a total of eight user selectable
traces including, four QuadTec wavelengths, conductivity, %B concentration based on method in use, pump
pressure, SIM pH, and SIM with any detector (fluorescence, refractive index).
Figure 2-1. QuadTec with the BioLogic DuoFlow System
USE WITH THE BIOLOGIC DUO-FLOW SYSTEMSYSTEM INSTALLATION AND SETUP
2-1
UNO Q1 COLUMN
AVR7-3 SAMPLE
INJECT VALVE
ICM MODULE
COLOR MONITOR
MIXER
FITTINGS TIGHTENER
(GREEN CLAMP)
AB
CONTROLLER
WORKSTATION
MOUSEKEYBOARD
MODEL 2128 FRACTION
QUADTEC DETECTOR
CONDUCTIVITY
MONITOR
COLLECTOR
2.1 INSTRUMENT CONTROL MODULE (ICM): FOR USE WITH THE DUOFLOW ONLY
The ICM translates the signal from the QuadTec detector and transmits it to the BioLogic DuoFlow
workstation. The ICM contains the following:
Front View
• Address setting: The dial should always be set to position 1
• Instrument bus connector: This connector is for System Cable 17 which connects to the BioLogic
Workstation and the Model 2128 Fraction Collector if used. (Refer to Figure 2-3.)
Rear View
• Serial Comm connector: This connector is for system cable 25 (QuadTec RS232 cable), which
connects to the QuadTec detector
• Power connector: This connector connects to the BioLogic DuoFlow workstation DC outlet with
system cable 26
Figure 2-2.
Instrument Control Module (ICM)
(front and rear views)
USE WITH THE BIOLOGIC DUO-FLOW SYSTEM SYSTEM INSTALLATION AND SETUP
2-2
INSTR. BUS INSTR. BUS
DEVICE
NUMBER
2
1
0
7
6
3
4
5
RX
TX
FRONT VIEW REAR VIEW
COMM
POWER
_ _ _
9...25 V
0.3A MAX
ON
2.2 ELECTRICAL CONNECTIONS TO THE DUOFLOW SYSTEM
The QuadTec communicates with the BioLogic DuoFlow system via the ICM module. To connect the
QuadTec to the DuoFlow system:
1. Use System Cable 25 to connect from the RS232 connector on the back of the QuadTec to the
COMM connector on the ICM module.
2. Use System Cable 26, the ICM power cord, to connect to the back of the workstation.
3. Use System Cable 17 to connect between an instrument bus connector on the ICM and an available
instrument bus connector on the workstation.
4. If you are using a Bio-Rad device with an instrument bus connector, such as the Model 2128 fraction
collector, use another system cable 17 to connect between the instrument bus connectors on the
Model 2128 and the ICM.
Figure 2-3. Connecting to the DuoFlow System
USE WITH THE BIOLOGIC DUO-FLOW SYSTEMSYSTEM INSTALLATION AND SETUP
2-3
COLOR MONITOR
CONTROLLER
MOUSE
SYSTEM
CABLE 17
SYSTEM CABLE 25
KEYBOARD
SYSTEM CABLE 26,
ICM POWER CORD
AVR7-3
INJECT
VALVE
WORKSTATION
SYSTEM CABLE 17
SYSTEM CABLE 17
MODEL 1327
CHART RECORDER
SYSTEM
CABLE 2
CONDUCTIVITY
MONITOR
SYSTEM
CABLE 4
MIXER
QUADTEC
DETECTOR
ICM MODULE
MODEL 2128
FRACTION COLLECTOR
2.3 PLUMBING CONNECTIONS TO THE DUOFLOW SYSTEM
The QuadTec has a PEEK flow cell that must be installed by the user. The detector fittings which have been
provided are standard HPLC 10-32 fingertight fittings. The BioLogic system uses 1/4-28 fittings.
To plumb the QuadTec to the BioLogic DuoFlow system:
• Plumb the detector after the column and before the conductivity monitor with the orange or green
PEEK tubing provided with the DuoFlow. The tubing length between the column and the detector
should be as short as possible.
• The flow is from the left to the right side of the instrument as shown in the picture below.
Figure 2-4. Plumbing to the DuoFlow System
USE WITH THE BIOLOGIC DUO-FLOW SYSTEM SYSTEM INSTALLATION AND SETUP
2-4
CONTROLLER AND
COLOR MONITOR
INSTRUMENT
BUS
MIXER
WORKSTATION
A B
AB
BUFFER A and B
WASTE
WASTE
COLLECT TO MODEL 2128
6
7
1
FRACTION COLLECTOR
2
INJECT
PORT
RINSE
5
4
3
AVR7-3
INJECT
VALVE
COLUMN
QUADTEC
DETECTOR
CONDUCTIVITY
MONITOR
2128
DIVERTER
VALVE
2.4 SYSTEM POWER UP
To power up the QuadTec Detector with the BioLogic DuoFlow system:
1. Important: Ensure that the BioLogic DuoFlow software application is not
running.
2. Turn on power to the DuoFlow workstation. The ICM power LED should light, since it receives power
from the workstation. If it doesn’t, ensure that the ICM power cable is connected to the workstation.
3. Power up the QuadTec using the power switch on the rear of the detector.
The QuadTec detector goes through a startup routine, self-test and lamp calibration routine.
4. If these routines are successful, the bottom left of the QuadTec screen will display ON. At this point,
the BioLogic DuoFlow software application may be launched.
5. When the DuoFlow Controller establishes communication with the QuadTec detector, the top left of
the QuadTec detector screen will show SLAVE. The BioLogic Controller will show a QuadTec
detector faceplate in the Manual screen. The system is then ready for operation.
2.5 SYSTEM OPERATION
When the QuadTec is connected to the BioLogic DuoFlow system, it is automatically recognized and its
functions are controlled from the Manual Control screen and the Run screen. These are discussed below.
2.5.1 Manual Screen
In the Manual screen (Figure 2-5), the QuadTec appears in its own control panel; the QuadTec controls are
listed below. The QuadTec Detector control panel can be toggled to the BioLogic standard UV monitor
control panel.
• Mode: To control the QuadTec from the DuoFlow system, select System mode. To control the
QuadTec from its front panel, select Local mode.
• Zero baseline: Resets the detector AU value to zero.
• On/Off: Turns the lamp on and off.
• Lamp: Indicates whether the lamp is Deuterium or Halogen.
• Range: Displays the wavelength range for the lamp in use. The range for Deuterium is 190–370 nm;
that for Halogen is 370–740 nm.
• Wavelength selection: Use the checkbox and up/down arrows to select each wavelength.
Note: Wavelengths selected in the Manual screen will not be automatically selected in the Method
Protocol Editor screen.
• Set: Implements the wavelength setting.
USE WITH THE BIOLOGIC DUO-FLOW SYSTEMSYSTEM INSTALLATION AND SETUP
2-5
Figure 2-5. Manual Control Screen, Showing the QuadTec Detector
USE WITH THE BIOLOGIC DUOFLOW SYSTEM SYSTEM INSTALLATION AND SETUP
2-6
BioLogic Duo-Flow - <user name> - <project name> - <method name>
File View Utilities Options
New
Method
Method
New
Edit
Run Browser Manual Setup Setup
Report
Window Help
wash
load12
Protocol
Run
Notes
PostRun
Log Settings
Gradient pump: F-10
Flow rate:
Inlet A:
Inlet B:
High
limit:
Low
limit:
1.00
100
0
1000
0
START
GP-pressure
Fraction collector
Mode: System Local Mode:
ml/min
Rack:
Rack 1 (128 tubes)
%
%
psi
psi
Start Tube:
End Tube:
Fraction size:
Tube number:
1
128
1.00
5
Volume left:
Set
0.15 ml
Advance
STOP START STOP
21
34
2.00
100% Buffer B
1.50
1.00
50
0.50
0.00
ml
Fractions
QuadTec Detector
System Local
Zero Baseline
Lamp
Range
Deuterium
190 - 370 nm
Wavelength Selection
280
260
214
405
AVR7-3 at port 4SVT3-2 at port 1 SV5-4 at port 2
I
PL21
Econo Gradient Pump
Mode:
OFF
Flowrate:
EGP B
Split
nm
nm
Flow
Direction
nm
nm
Set
START STOP
AVR9-8 at port 5
I
82
37
6
4
5
500.0
400.0
300.0
200.0
100.0
0.0
System Local
0.100
0
0.00
ml/mir
%
%
Set
QuadTec-3 (214 nm)
02 4 6 8
AU
QuadTec
WL1 - 280nm
4.000 AU
WL2 - 260nm
-0.00101 AU
WL3 - 214nm
-0.00100 AU
Gradient Pump: F-10 UV Conductivity
1.00ml/min 0 %B
438 psi -0.00278 AU 379 mS/cm
Clear all trace data, start chromatogram from time = 0
Minutes
WL4 - 405nm
-0.00100 AU
Econo Gradient
Pump
10
mS/cm
Flow Rate EGP %B
0.100 ml/min
0 %B
1.907 AU 0.00 pH
Clear
Traces
Split
0%
SIM1/pHSIM1/SIG
The traces shown on the Manual screen chromatogram and printed report are chosen in the Chromatogram
Settings window, shown in Figure 2-6. From the system toolbar, select the Settings button to display the
Manual screen Chromatogram Settings window. Select the traces from the Trace Device drop-down menu.
Select up to eight traces. Select the Visible checkbox to indicate whether or not the trace is to be visible on
the screen.
Figure 2-6. Manual Screen Chromatogram Settings
To set the QuadTec detector’s Time Constant, select Manual Setup from the Options drop-down menu. In
the Manual Setup screen, shown below, enter the QuadTec Time Constant. The Time Constant value
smooths the signal with a time filter. The default value of 1.0 seconds is suitable for most applications.
Figure 2-7. Manual Setup Screen
USE WITH THE BIOLOGIC DUO-FLOW SYSTEMSYSTEM INSTALLATION AND SETUP
2-7
Chromatogram Settings
Settings for the Manual Mode
Y-axis settings
Trace Device Visible UnitsBaseline
UV
UV
Conductivity
x
SIM1/Signal
SIM1/pH
x
SIM2/Signal
x
SIM2/pH
GP-pressure
QuadTec-1 (220 nm)
x
QuadTec-2 (280 nm)
x
QuadTec-3 (250 nm)
x
QuadTec-4 (595 nm)
x
%Buffer B
x
10
-0.000
Axis Max
2.000
AU
Ok
Cancel
Default
Manual Setup
EGP Split Time Period
0.0 sec.
Select SIM
QuadTec Time Constant
1.0 sec.
Edit Signal Input Module 1 - Signal
SIM 1
SIM 2
Device Output Range
Units Range:
(Volts):
Gradient Pump : F-10 Purge FlowRate
10.00 ml/min
VoltUnits:
Minimum
0.000
0.000
Maximum
1.000
1.000
Ok
Cancel
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