Bio-Rad BioLogic QuadTe Detector and Components User Manual

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BioLogic

uadTec™

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

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

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

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

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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

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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

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FLOW CELL

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.

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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)

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)

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2-2

INSTR. BUS INSTR. BUS

DEVICE

NUMBER

RX TX

FRONT VIEW REAR VIEW

COMM

POWER

_ _ _

9...25 V

0.3A MAX

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

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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

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

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2-4

CONTROLLER AND

COLOR MONITOR

INSTRUMENT

BUS

MIXER

WORKSTATION

A B

BUFFER A and B

WASTE

COLLECT TO MODEL 2128

FRACTION COLLECTOR

INJECT PORT RINSE

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.

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Figure 2-5. Manual Control Screen, Showing the QuadTec Detector

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BioLogic Duo-Flow - <user name> - <project name> - <method name>

File View Utilities Options

New

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

1000

START

GP-pressure

Fraction collector

Mode: System Local Mode:

ml/min

Rack:

Rack 1 (128 tubes)

psi

Start Tube:

End Tube:

Fraction size:

Tube number:

128

1.00

Volume left:

Set

0.15 ml

Advance

STOP START STOP

2.00 100% Buffer B

1.50

1.00

0.50

0.00

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

PL21

Econo Gradient Pump

Mode:

OFF

Flowrate:

EGP B

Split

Flow Direction

Set

START STOP

AVR9-8 at port 5

500.0

400.0

300.0

200.0

100.0

0.0

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0.100

0.00

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Set

QuadTec-3 (214 nm)

02 4 6 8

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

mS/cm

Flow Rate EGP %B

0.100 ml/min

0 %B

1.907 AU 0.00 pH

Clear

Traces

Split

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

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Chromatogram Settings

Settings for the Manual Mode

Y-axis settings

Trace Device Visible UnitsBaseline

Conductivity

SIM1/Signal SIM1/pH

SIM2/Signal

SIM2/pH GP-pressure QuadTec-1 (220 nm)

QuadTec-2 (280 nm)

QuadTec-3 (250 nm)

QuadTec-4 (595 nm)

%Buffer B

-0.000

Axis Max

2.000

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

Maximum

1.000

Cancel

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