Operation Manual RI 2031aa

P/N: 0302-0555A

MODEL RI-2031 INTELLIGENT REFRACTIVE INDEX

DETECTOR

INSTRUCTION MANUAL

i

Safety Considerations

To ensure operation safety, this instrument must be operated correctly and maintained regularly according to schedule. Carefully read to fully understand all safety precautions in this manual before operating the instrument. This manual denotes precautions against actions that can result in hazardous situations or equipment damage by using the signal words WARNING, CAUTION, and Note.

(1) Safety symbols

Instruction manual symbol. If the product is marked with this symbol, refer to the instrument manuals to protect the instrument against damage.

WARNING A WARNING indicates a potentially hazardous situation which, if not avoided, could result in death or serious injury.

CAUTION A CAUTION indicates a potentially hazardous situation which, if not avoided, may result in minor or moderate injury. It may also be used to alert against unsafe practices.

Do not proceed beyond a WARNING or CAUTION notice until you understand the hazardous conditions and have taken the appropriate steps.

Note A Note provides additional information to aid the operator in obtaining optimal instrument performance.

Pressurized, hazardous solvents are used in high-performance liquid chromatography. Always follow the proper laboratory procedures to ensure operator safety. Always wear goggles, gloves and protective clothing when operating the instrument, especially when injecting a sample and opening valves.

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(2) Warning Labels Warning labels are attached at several locations on this instrument. Do not remove, deface or damage the warning labels. If a warning label peels off the instrument or becomes illegible, contact your local JASCO distributor for a replacement label. Be sure to indicate the part number (P/N) on the label.

(1) Fuse and Ground Warning Label (P/N: 0822-0163A)

Figure 1 Figure of the side view

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Regulatory Statements CE Notice Marking by the symbol indicates compliance of this JASCO system to the EMC (Electromagnetic Compatibility) and Low Voltage Directives of the European Community. This symbol indicates that this JASCO system meets the relevant basic safety and health requirements of the EC Directive based on the following technical standards: • EN55011 -- ”Limits and Methods of Measurement of Radio Interference

Characteristics of Information Technology Equipment.” -- Group 1, Class A.

• EN61000-6-1 -- "Electromagnetic compatibility (EMC) Part 6-1:Generic

standards – Immunity for residential, commercial and light-industrial environments"

• IEC61000-3-2 -- "Electromagnetic compatibility (EMC) Part 3-2:Limits – Limits for harmonic current emissions (equipment input current up to and including 16A per phase) "

• IEC61010-1 -- "Safety requirements for electrical equipment for measurement, control and laboratory use – Part 1:General requirements"

• A "Declaration of Conformity" in accordance with the above standards has been made and is on file at JASCO EUROPE srl, Via Confalonieri 25, 22060 CREMELLA (LC), Italy.

FCC Statement (for USA only) Federal Communications Commission Radio Frequency Interference Statement

Warning

This is a Class A product. In a domestic environment this product may cause radiointerference, in which case the user may be required to take adequate measures.

Warning This equipment generates, uses, and can radiate radio frequency energy. If it is not installed and used in accordance with the instruction manual, it may causeinterference to radio communications. It has been tested and found to comply with the limits for a Class A computing device pursuant to Part 15 of FCC Rules, which are designed to provide reasonable protection against such interference when operated in a commercial environment. Operation of this equipment in a residential area is likely to cause interference, in which case the user at his own expense will be required to take whatever measures may be required to correct the interference.

iv

Preface

This instruction manual serves as a guidebook for using this instrument. It is intended to instruct first-time users on how to properly use the instrument, and to serve as a reference for experienced users.

Before using the instrument, please read this instruction manual carefully, and make sure that the contents are fully understood. This manual should be easily accessible to the operator at all times during instrument operation. When not using the instrument, keep this manual stored in a safe place. Should this instruction manual be lost, order a replacement from your local JASCO distributor.

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Installation Conditions

To ensure safe operation, the following recommendations should be observed:

(1) Do not operate the instrument under voltage fluctuations exceeding 10% of the recommended line voltage. Large fluctuations may cause the instrument to fail.

(2) Use a three-pronged electrical outlet with a ground. When only a two-pronged socket is available, use an adapter and be sure to connect the ground wire of the adapter.

(3) Operate the instrument under a temperature range of 10 ∼ 30°C.

(4) Operate the instrument under a humidity range of 35 ∼ 85% (RH). If ambient humidity exceeds 85% (RH), water vapor may deteriorate optical components. If possible, install the instrument in a location having a humidity of 60% or lower.

(5) Operate the instrument under an atmospheric pressure of 750 ∼ 1060 hPa.

(6) Avoid strong magnetic fields and sources of high-frequency waves. The instrument may not function properly when near strong magnetic fields or high-frequency wave sources.

(7) Avoid vibrations caused by vacuum pumps, electric motors, processing equipment and machine tools.

(8) Avoid dust and corrosive gas. Do not install the instrument in a location where it may be exposed to dust, especially in locations exposed to outside air or ventilation outlets that discharge dust particles.

(9) Do not install the instrument in a location where it may be exposed to direct sunlight.

(10) Do not install the instrument in a location where it may be directly exposed to the air current from an air conditioner or heater, as such a location may inhibit stable measurement.

Note: The above conditions do not ensure optimal performance of this instrument.

vi

Maintenance

Consult your local JASCO distributor regarding maintenance. In addition, contact your local JASCO distributor when transporting the instrument.

Replacement parts can be ordered according to part number from your local JASCO distributor. When the part number is not known, inform your local JASCO distributor of the part name, instrument model name and its serial number.

Notices

(1) Jasco shall not be held liable, either directly or indirectly, for any consequential damage incurred as a result of product use.

(2) Software prohibitions:

Copying of software or related materials for purposes other than backup is prohibited.

Transfer or authorization of the use of Jasco software to or by a third party is prohibited.

Disclosure of confidential information related to Jasco software is prohibited.

Modification of Jasco software is prohibited.

Use of Jasco software on multiple workstations or terminals, through a network or through any other means, is strictly prohibited. (This does not apply to entities with the network license contract)

(3) The content of this manual is subject to change without notice in accordance with product improvements.

(4) Unauthorized copying of this manual is prohibited.

(5) This manual shall not be used to guarantee or copyright industrial rights or other rights.

(6) Company and product names listed herein are trademarks or registered trademarks of their respective owner.

vii

Warranty

This product is warranted for a period of one year from the date of delivery. If any defects should occur in the product during this period of warranty, JASCO will repair or replace the defective part(s) or product free of charge.

This warranty does not apply to defects as a result of the following:

(1) USE FOLLOWING IMPROPER OR INADEQUATE INSTALLATION.

(2) IMPROPER OPERATION.

(3) MOVEMENT, MODIFICATION, OR REPAIR BY PERSONS OTHER THAN AUTHORIZED JASCO PERSONNEL.

(4) USE OF PARTS OTHER THAN THOSE THAT ARE AUTHORIZED BY JASCO.

(5) INORDINATELY RAPID DETERIORATION DUE TO THE USE OF CORROSIVE SOLVENTS OR SAMPLES.

(6) NATURAL DISASTERS SUCH AS FIRES, WATER DAMAGE, OR EARTHQUAKES.

In addition, this warranty does not cover:

(7) CONSUMABLE PARTS OR PARTS THAT HAVE A SEPARATE WARRANTY OR A WARRANTY PERIOD THAT IS DIFFERENT THAN THAT SPECIFIED ABOVE.

The warranty period for all parts and repairs supplied under this warranty expires with the warranty period of the original product.

Jasco Corporation

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Table of Contents Safety Considerations ...................................................................i Regulatory Statements................................................................iii Preface..........................................................................................iv

Installation Conditions .................................................................v

Maintenance .................................................................................vi Warranty ......................................................................................vii Table of Contents.........................................................................ix

1. Outline and Specifications...................................................1

1.1 Outline .................................................................................................... 1

1.2 Specifications ........................................................................................ 2

2. Part Names and Functions ..................................................4

2.1 Front panel ............................................................................................. 4

2.2 Operation panel ..................................................................................... 7

2.3 Back panel.............................................................................................. 8

3. Power On/Off and Self-diagnostics...................................10

3.1 Power on and self-diagnostics ........................................................... 10

3.2 Power off .............................................................................................. 11

4. Operations in Normal Operation Mode .............................12

4.1 Parameter changes.............................................................................. 12 4.1.1 Range and response changes..................................................... 12 4.1.2 Temperature control ON/OFF and temperature setting ............ 15 4.1.3 Output polarity change method ([SHIFT][6]) .............................. 15

4.2 Purge valve open/close (Solvent replacement in the reference cell) ............................................................................................................. 17

4.3 Auto-zero operations........................................................................... 18

4.4 Error and warning messages during monitoring.............................. 18

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4.5 Measurement of concentrated sample (Refractive index range shifting................................................................................................. 19

5. [SHIFT] Key Operations .....................................................21

5.1 Operation summary............................................................................. 21

5.2 Preamplified output display ([SHIFT][1]) ........................................... 22

5.3 Auto-zero operation setting ([SHIFT][3])............................................ 23

5.4 Zero point shift for integrator output ([SHIFT][4])............................. 24

5.5 Lamp operation time ([SHIFT][5]) ....................................................... 26

5.6 Signal filtering mode ([SHIFT][7]) ....................................................... 27

5.7 Other settings ([SHIFT][MARK]) ......................................................... 27 5.7.1 Marker voltage/polarity changes................................................. 29 5.7.2 Setting to continue/stop self-diagnostics when an error occurs................................................................................................................ 30

6. Program Mode Operations.................................................32

6.1 Operation summary............................................................................. 32

6.2 Switching between normal operation mode and program mode .... 32

6.3 File number setting (file loading) ....................................................... 33

6.4 Program editing ................................................................................... 34 6.4.1 Initial parameter editing ............................................................... 34 6.4.2 Time program editing................................................................... 35

6.5 Time program operation...................................................................... 40 6.5.1 Time program execution.............................................................. 40

7. MEASURE RANGE Setting.................................................41

8. Maximum Flow Rate Changing at the Front Panel...........43

8.1 “ ~ 10mL/min” tubing arrangement and solvent replacement method for reference cell................................................................................. 43

8.2 “10 ~ 50mL/min” tubing arrangement and solvent replacement method for reference cell................................................................... 44

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9. Maintenance........................................................................45

9.1 Error messages and countermeasures.............................................. 45 9.1.1 Errors during self-diagnostics after power on........................... 45 9.1.2 Self-diagnostics during operation .............................................. 46 9.1.3 Error message meanings and countermeasures....................... 46

9.2 Flow cell bubble removal .................................................................... 48 9.2.1 Baselines when bubbles are generated ..................................... 48 9.2.2 Bubble removal method 1 (high flow rate fluid pump).............. 50 9.2.3 Bubble removal method 2 (syringe fluid injection) ................... 50 9.2.4 Bubble removal method 3 (covering tubing outlet) ................... 50 9.2.5 Bubble prevention method .......................................................... 50

9.3 Flow cell cleaning method .................................................................. 51

9.4 Calibration method .............................................................................. 52 9.4.1 Tubing ........................................................................................... 53 9.4.2 Standard solution ......................................................................... 53 9.4.3 Replacement of tubing system solvent with water.................... 54 9.4.4 Calibration method....................................................................... 54 9.4.5 Calibration constant confirmation and setting .......................... 56 9.4.6 Alternative calibration method.................................................... 57

9.5 Lamp maintenance time setting ......................................................... 58

9.6 Serial number setting .......................................................................... 59

9.7 Power Fuse replacement..................................................................... 60

10. Appendix.............................................................................62

10.1 Operation Theory............................................................................... 62 10.1.1 Operation theory of deflection differential refractometers ..... 62 10.1.2 Optical system............................................................................ 63 10.1.3 Signal processing method......................................................... 65 10.1.4 Flow line ...................................................................................... 66 10.1.5 Internal volume of tubing........................................................... 68

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1. Outline and Specifications

1.1 Outline

The JASCO RI-2031 Intelligent Refractive Index Detector is an HPLC detector which offers unprecedented versatility, performance, and stability. The RI-2031 features the following:

High stability

High stability is obtained through an optical system that is not easily influenced by temperature, an efficient thermal exchanger, and a digital signal processing system.

Wide dynamic range

10 times larger dynamic range than an ordinary HPLC refractive index detector.

Maximum flow rate of 50mL/min.

Maximum flow rate is 50mL/min. (Solvent: H2O) Wide dynamic range and Extended Maximum flow rate are applied to the semi-preparation.

Easy operation

Parameters can be easily set using the LCD display, function keys, and numeric keys. Self-diagnostics results and warning messages are also displayed, which allows the condition of the instrument to be accurately monitored. Optical Zero can be performed precisely using the easy key operation.

Automatic analysis capability

The internal time program allows automatic switching of sensitivity range, response and other functions. Using LC-Net, parameters can be set from the Model HSS-2000 System without an interface.

Compact design

The front panel dimensions are the same as other JASCO 2000 series HPLC equipment (pump, UV detector), which facilitates a neat stacking arrangement.

Safety measures An internal fluid leak sensor is installed. When a leak from the flow cell occurs, a signal is output and the pump can be stopped.

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1.2 Specifications

Model name: RI-2031 Intelligent Refractive Index Detector

Measurement system: Deflection type

Refractive index range: 1.00 ~ 1.75

Measurement range settings: 1/4, 1/2, 1, 2, 4, 8, 16, 32, 64, 128, 256, 512 x

10-6RIU/full scale (“MEASURE RANGE”: STD)

1/4, 1/2, 1, 2, 4, 8, 16, 32, 64, 128, 256, 512 x

10-5RIU/full scale (“MEASURE RANGE”: WIDE)

Linearity: 5.0 x 10-4 RIU (“MEASURE RANGE”: STD)

5.0 x 10-3 RIU (“MEASURE RANGE”: WIDE)

<Condition> H2O

Noise: 0.2 x 10-8RIU

<Condition> H2O, Flow stopped

Cell capacity: 10µL

Maximum usable flow rate: 10mL/min. (“ ~ 10mL/min.” tubing)

50mL/min. (“10 ~ 50mL/min.” tubing)

<Solvent> H2O

Maximum pressure: 0.1MPa. (“ ~ 10mL/min.” tubing)

0.3MPa. (“10 ~ 50mL/min.” tubing)

Temperature control: PID control by heater,

Setting range: 0 ~ 45°C in 1°C increments.

Control range: Room temperature +10°C ~ Room temperature +25°C

Zero adjustment: Automatic Optical and electronic auto-zero functions.

Integrator output zero positioning

Reference selection: Electromagnetic valve control

(“ ~ 10mL/min.” tubing)

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Time program: Parameters that can be set are PURGE ON/OFF, AUTO ZERO, RANGE, and RESPONSE. Range from 0.1 to 999.9min.

Response FAST, STD, SLOW (TIME ACCUME, DIGITAL FICTER)

Input and output signals: Recorder output: 10mV/FS

Integrator output: 5.0 x 10-4RIU/1V(SID)

5.0 x 10-3RIU/1V(WIDE)

Marker and leak output: 1 circuit each

Marker, auto-zero, program reset/start: 1 circuit each

LC-Net control: 1 circuit

Liquid wetted material: Quartz, fluoride resin, stainless steel

(SUS316)

Dimensions and weight: 150(W) x 470(D) x 150(H) mm, excluding

Protrusions.

Approximately 13kg

Power requirements: AC100 ~ 240V, AC, 50/60Hz, 50VA max.

Ambient temperature: +10 ~ +35°C for operation

-30 ~ +60°C for storage

* Specifications are subject to change without notice.

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2. Part Names and Functions

2.1 Front panel

POWER Switch

Operation Panel

Solvent Inlet and Outlet

Figure 2.1 Front Panel

Name Function Solvent Inlet and Outlet “1” ~ “6” Operation panel POWER switch

Sample and Reference In and out connections Refer to Figure 2.2 and Figure 2.3. Panel containing control keys, parameter edit keys, and LCD display showing operating conditions, parameters and various messages. Refer to Section 4 and 5. Power switch for the instrument

Note: Suitable tubing for the Solvent Inlet and Outlet connections must be selected according to maximum flow rate. Before connecting tubing, refer to Figure 2.2. and Figure 2.3. “ ~ 10mL/min”. tubing is supplied with the instrument.

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Note: For a “10 ~ 50mL/min” tubing arrangement, the refractive index signal output (and display) polarity becomes negative. Change the polarity setting from plus (+) to minus (-) using the polarity setting function. (Refer to 4.1.3 polarity setting.)

Figure 2.2 “ ~ 10mL/min” flow rate tubing connections

3

4 5 6

2 1 Sample Out (waste)

Sample IN (from column)

Tubing with OD=1.6mm ID= 0.8mm

Flow rate: ~ 10mL/min.

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Figure 2.3 “10 ~ 50mL/min” flow rate tubing connections

Flow rate: 10 ~ 50mL/min.

Tubing OD=1.6mm, ID=0.8mm

Replace solvent in the reference cell by sucking the mobile phase solvent. Keep the syringe connected during analysis in order to maintain solvent in the reference cell and to facilitate solvent replacement.

Flow rate: 10 ~ 50mL/min.

Tubing OD=1.6mm ID=0.8mm (Same tubing as used to connect ports No.1-2 or ports No.4-5. for the “ ~ 10mL/min” tubing arrangement.

Tubing OD=1.6mm ID=0.8mm

Sample Out

Sample IN

Reference IN

Reference Out

1

Sample Out

Sample IN

Mobile phase

2 3

4 5

1 2 3

4 5 6

6

7

2.2 Operation panel

Figure 2.4 Operation Panel

Name Function LCD screen Displays operation conditions, setting values, error

messages and other information. [PRGM RUN] Used to start and stop the time program. Operation status

is indicated by the lamp to the upper left of the button. The lamp is lit during program operation.

[AUTO ZERO] Sets the refractive index unit value (RIU) to zero. [MARK] Outputs a marker signal from the recorder output terminal. [MONIT] Used to return to the monitor screen from other screens

and to switch between monitor screens. [SHIFT] Used in combination with other keys to set parameters that

are not frequently changed. [PURGE] Executes solvent replacement (ON/OFF) for the reference

cell. [PRGM] Switches between normal and time program mode. [0] ~ [9], [.] Numeric keys. Keys 1 ~ 5 are used for special functions in

addition to their numeric function. The letters on these keys describe the special functions. In other words, when editing parameters, these keys are used to call up the relevant functions. Refer to sections 4.1.1 and 6.4.2 for details.

[ ][ ] keys Used to edit parameters and change screens. [CLEAR] Used to erase incorrect parameter input and clear error

messages. [EDIT/ENTER] Used to edit parameters.

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2.3 Back panel

Input/Output Terminals LC-Net Connectors

RS-232C Terminal

Fuses

AC input

Ground Terminal

Figure 2.5 Back Panel

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Name Function Input and output terminals Signal output terminals REC+/- G(GND)- INT+/- G(GND) MARK OUT +/- LEAK OUT +/- Signal input terminals MARK IN+ GND- A/Z IN+ GND- PRGM RST/ST+ GND- Cooling fan AC input Power fuses Ground terminal LC-Net connector RS-232C terminal

Recorder output terminal (Output level determined by RANGE setting) Integrator output terminal (Fixed scale of 5x10-4RIU/1V(“MEASURE RANGE”: STD), 5x10-3RIU/1V(“MEASURE RANGE”: WIDE). Marker output terminal (Normally open, contact type output is closed when [MARK] key is pressed or MARK IN is input) Fluid leak warning output terminal (Normally open, contact type output is closed when a fluid leak from the flow cell occurs) (Note) The internal fluid leak sensor can only detect a leak of buffer solution. Marker input terminal (Marker is added on the recorder output when the contact point is closed) Auto-zero input terminal (Recorder and integrator outputs are zeroed when contact type input is closed) Time program input terminal (Time program is reset and immediately started when contact type input is closed) Used for cooling the internal components of the instrument Accepts the power cable. Protects the instrument from excessive currents. Grounds the instrument. This is not the ground terminal for input and output signals. Input and output terminal for the LC-Net. Used when controlling the instrument via a RS-232C cable (optional).

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3. Power On/Off and Self-diagnostics

3.1 Power on and self-diagnostics

Turn the power switch, located at the lower left of the front panel, to the ON position. The self-diagnostics program is automatically executed. The program examines the following items:

ROM CHECK (Memory)

RAM CHECK (Memory)

DC POWER (Direct current power source)

BACK UP (Memory backed up by battery)

TEMP SENSOR (Temperature sensor)

LAMP CHECK (Lamp energy)

LAMP O.P TIME (Lamp operating time)

ZERO GLASS DRV. (Optical zero glass driver)

An error message will appear if a problem is detected.

Note: When an error occurs, diagnostics will either stop immediately or continue to the next item depending on the instrument settings (refer to section 5.7.2).

The screen on the left in Figure 3.1 will appear when diagnostic tests are complete. Press the [MONIT] key to switch to the screen on the right in Figure 3.1. The heater temperature is displayed in the screen on the right. When the [MONIT] key is pressed again, the display will return to the screen on the left. Normal operation mode consists of these two screens.

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NORM 64 STD

-0.1 + ON

MODE RANGE RESPONSE

RIU(x10 ) POLARITY PURGE

Mode display

(Normal operation mode) Sensitivity

(64)

Response (standard)

Polarity

(Positive)

Purge valve status(ON)

-6

TEMP: xx.x/yy

-0.1 + ON

MODE RANGE RESPONSE

RIU(x10 ) POLARITY PURGE -6

[MONIT]

Current Optical bench temperature/setting temperature

Temperature control markblinkswhen heating

Normal operation mode monitor screen(1) Normal operation mode monitor screen(2)

°C

Current refractive index

(-0.1)

Figure 3.1 Normal Operation Mode Screens

3.2 Power off

Turn the power switch, located at the lower left of the front panel, to the OFF position. The range, response, polarity, temperature, and time program set at this time will be recorded in C-MOS RAM (memory backed up by battery). These values will be restored when the power is turned on again.

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4. Operations in Normal Operation Mode The dynamic measurement range of the RI-2031 is 10 times larger than ordinary HPLC refractive index detectors. The dynamic measurement range of the RI-2031 depends on the “MEASURE RANGE” setting. When a highly concentrated sample is analyzed, set the “MEASURE RANGE” to WIDE. Refer to section 7.

The maximum flow rate of the RI-2031 depends on the tubing connections of the front cell panel. The “ ~ 10mL/min.” tubing connections are for conventional analysis and the “10 ~ 50mL/min” tubing connections are for semi-preparative analysis. Tubing connection configurations are described in section 8.

4.1 Parameter changes

4.1.1 Range and response changes

After the power is turned on and the self-diagnostic tests are complete, the normal operation mode monitor screen (1) (left in Figure 4.1) will appear. Range and response settings can be changed here.

Note: Use the [MONIT] key to switch between monitor screens.

NORM 64 STD

-0.1 + ON

MODE RANGE RESPONSE

RIU(x10 ) POLARITY PURGE

Mode display

(Normal operation mode) Sensitivity

(64)

Response (standard)

Polarity

(Positive)

Purge valve status(ON)

-6

TEMP: xx.x/yy

-0.1 + ON

MODE RANGE RESPONSE

RIU(x10 ) POLARITY PURGE -6

[MONIT]

Current optical bench temperature/setting temperature

Temperature control markblinkswhen heating

Normal operation mode monitor screen(1) Normal operation mode monitor screen(2)

°C

Current refractive index

(-0.1)

Figure 4.1 Normal Operation Mode Screens

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Input range:

1/4, 1/2, 1, 2, 4, 8, 16, 32, 64, 128, 256, 512, S (Short) x10-6RIU /10mV

(“MEASURE RANGE”: STD)

1/4, 1/2, 1, 2, 4, 8, 16, 32, 64, 128, 256, 512, S (Short) x10-5RIU /10mV

(“MEASURE RANGE”: WIDE)

Response input range: FAST, STD, SLOW

(1) Change method 1

Flashes

Normal operation mode monitor screen(1)

Sensitivity input screen

Values in parentheses are after changes.

Display the desired value [ ][ ][EDIT/ENTER]

NORM 64(16) STD(FAST)-0.000 + ON

NORM 64 STD

-0.000 + ON

[EDIT/ENTER]

Response rate input screen

Display the desired value

[MONIT]

NORM 16 STD

-0.000 + ON

Flashes

[ ][ ][EDIT/ENTER]

Figure 4.2 Change Method (1)

Note: If an incorrect value is entered accidentally, press the [CLEAR] key before pressing the [EDIT/ENTER] key then enter the value again.

Change method 2

When the [RANGE/4] (range) key or the [RSPNS/5] (response) key are pressed, the corresponding parameter will flash and the value can be changed directly.

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Sensitivity flashes

[RANGE/4] [ ][ ][EDIT/ENTER]

NORM 64 STD

-0.000 + ON

Response flashes

[RSPNS/5]

Sensitivity change Response change

NORM 64(16) STD

-0.000 + ON

NORM 64 STD(FAST)

-0.000 + ON

NORM 64 STD

-0.000 + ON

[ ][ ][EDIT/ENTER]

Figure 4.3 Change Method (2)

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4.1.2 Temperature control ON/OFF and temperature setting

When the normal operation monitor screen (2) (right in Figure 4.1) is displayed, temperature control can be turned ON/OFF and temperature can be set as shown the operations in Figure 4.4.

Input range: 0 ~ 45°C (Operating temperature range: Room temperature + 10°C ~ Room temperature +25°C)

0: OFF (Temperature control not active) Note: During heating operation, the mark at the top right of the monitor

screen will blink. Blinking speed is related to heater power. When 0 is set (control OFF), the mark belong to stop moving.

Flashes


Temperature input screen

Values in parentheses are after changes.

Values input[zz][EDIT/ENTER]

[EDIT/ENTER][MONIT]

TEMP: xx.x/yy(zz) °C

-0.0 + ON

TEMP: xx.x/yy °C

-0.0 + ON

Note:Temperature control will be turned off when 0 is input

Figure 4.4 Temperature Control ON/OFF and Temperature Setting

4.1.3 Output polarity change method ([SHIFT][6])

This operation changes the output polarity of the signal. When “10 ~ 50mL/min.” tubing is used, or negative peak heights exceed the integrator input range, polarity should be set to negative (-).

4.1.3.1 “10 ~ 50mL/min.” tubing arrangement

When “10 ~ 50mL/min.” tubing is used, because of the optical arrangement, the refractive index signal output decreases when the actual refractive index increases. Therefore, the polarity should be set to negative (-) in order to correct the relation between the actual refractive index and the refractive index signal output (for both the recorder and the integrator).

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When the actual negative peak signal exceeds -5.0x10-4RIU, polarity should be set to positive (+). Refer to Table 4.2 (“10 ~ 50mL/min.” tubing connections)

4.1.3.2 Negative peak signal exceeds the integrator input range

The allowable input voltage range of a current integrator is, in general, -10mV ~ +1V. When the negative peak signal is within 5x10-5 RIU (“MEASURE RANGE”: STD) or 5x10-4 RIU (“MEASURE RANGE”: WIDE), integrator output can be prevented from falling below –10mV using the zero point shift function. Refer to section 5.4.

When the negative peak signal exceeds this range, polarity should be changed. The actual allowable refractive index range is shown in Table 4.1 and Table 4.2. Change polarity settings, according to these tables.

Table 4.1 “ ~ 10mL/min.” tubing arrangement (*shifted by 100mV)

MEASURE RANGE polarity without zero point shift* with zero point shift*

STD + -5.0×10-6~+5.0×10-4 RIU -5.0×10-5~+4.5×10-4 RIU

STD - -5.0×10-4~+5.0×10-6 RIU -4.5×10-4~+5.0×10-5 RIU

WIDE + -5.0×10-5~+5.0×10-3 RIU -5.0×10-4~+4.5×10-3 RIU

WIDE - -5.0×10-3~+5.0×10-5 RIU -4.5×10-3~+5.0×10-4 RIU

Table 4.2 “10 ~ 50mL/min.” tubing arrangement (*shifted by 100mV)

MEASURE RANGE polarity without zero point shift* with zero point shift*

STD - -5.0×10-6~+5.0×10-4 RIU -5.0×10-5~+4.5×10-4 RIU

STD + -5.0×10-4~+5.0×10-6 RIU -4.5×10-4~+5.0×10-5 RIU

WIDE - -5.0×10-5~+5.0×10-3 RIU -5.0×10-4~+4.5×10-3 RIU

WIDE + -5.0×10-3~+5.0×10-5 RIU -4.5×10-3~+5.0×10-4 RIU

When polarity is set to negative (-) with a “ ~ 10mL/min” tubing arrangement, the screen display and output (for both the recorder and the integrator) will change as shown in Figure 4.5. When the actual refractive index decreases, the index (RIU) value on the screen will increase and so will output. Changes in peak appearance and display values caused by polarity differences are shown in Figure 4.5 and key operations are shown in Figure 4.6.

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0 0 0

RIU:-20.0

RIU:20.0

RIU:10.0

RIU:-10.0

Polarity:Positive Polarity:NegativeActual refractive index change

－

＋

Figure 4.5 Changes in Peak Appearance and Display Values Caused by Polarity Differences

POLARITY

+(-)

Input screen

Flashes

[MONIT] [SHIFT] [5]

Output polarity change screen

[EDIT/ENTER]

(-)is the value after the change.[ ][ ][EDIT/ENTER]

NORM 64 STD

-0.0 + ON

POLARITY

+


Figure 4.6 Output Polarity Change

Note: The marker output polarity can also be changed (refer to section 5.7.1).

4.2 Purge valve open/close (Solvent replacement in the reference cell)

This section explains the procedures to open and close the purge valve. When the valve is opened during fluid pumping (Screen display: ON), solvent flows to the cells on both the sample and reference side. When the valve is closed (Screen display: OFF), solvent only flows to the sample side. When replacing solvent or removing air bubbles in the flow cell, open the valve to pass solvent through the reference cell. Close the valve during analysis. The operations are shown in Figure 4.7.

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Normal operation mode monitor screen

NORM 64 STD

-0.0 + ON(OFF)

TEMP: xx.x/yy °C

-0.0 + ON(OFF)

[MONIT]

PURGE ON

1:ON 0:OFF

[PURGE]

[1]

[0] :Opens the purge valve and changes the status to ON.

Purge valve open/close screen

[MONIT]

Current purge valve status

:Closes the purge valve and changes the status to OFF.

Figure 4.7 Purge Valve Open/Close Operations

Note: When using “10 ~ 50mL/min” tubing connections, draw up the mobile phase solvent using a syringe in order to replace solvent in the reference cell. Replacement of solvent in the reference cell can be performed regardless of the PURGE VALVE setting.

4.3 Auto-zero operations

When the [AUTO ZERO] key is pressed, the current refractive index value is set as 0. In other words, the recorder and integrator output are set to 0.

Note: When a signal is input into MARK IN on the back panel, the auto-zero operation can be set to occur automatically. (Refer to section 5.3)

Note: When executing optical auto zero, press the [SHIFT][AUTOZERO] key. (Refer to section 5.2)

4.4 Error and warning messages during monitoring

The error and warning messages that may occur during monitoring are shown in Table 4.3.

When an error message is displayed, press the [CLEAR] key to return to the

19

monitor screen. If the problem continues, the same message will appear again after approximately 20 seconds. Refer to section 9.1 for countermeasures.

Table 4.3 Error Messages and Their Meanings Error message Meaning TROUBLE LEAK IN CELL TROUBLE OVER TEMP TROUBLE ZERO GLASS DRV WARNING INTENSITY POOR WARNING OUT OF RANGE WARNING LAMP MAINT TIME

Fluid is leaking from the flow cell Heater is overheating Zero glass drive not move Light intensity has deteriorated Optical zero adjustment is not correct Lamp maintenance time has been exceeded

4.5 Measurement of concentrated sample (Refractive index range shifting when the “MEASURE RANGE” setting is STD)

When “MEASURE RANGE” is set to STD, the linear dynamic range of refractive index, with properly adjusted optical zero, is +/-5.0x10-4RIU. Although this range cannot be expanded, it can be shifted in one direction. For example, the allowable refractive index range can be shifted to -3.0x10-4 ~ +7.0x10-4RIU.

The method for shifting the range is described below.

(1) Display the preamplified output screen (refer to section 5.2) and press the [ ][ ] keys so that DIFF is within 0±0.03

(2) Return to the monitor screen.

(3) Press the [AUTO ZERO] key to set the RIU value to 0.

(4) Display the preamplified output screen and press the [ ][ ] keys until the RIU value reaches -300.0±30(x10-6RIU) (this assumes the output polarity is set to positive).

(5) Again press the [AUTO ZERO] key to set the RIU value to 0. As long as optical zero adjustment is not executed, the allowable refractive index range will be -3.0x10-4 ~ 7.0x10-4RIU.

Note: The refractive index range resulting in proper integrator output is +/-5.0x10-4RIU, taking into consideration the polarity setting. When this

20

range is exceeded, use the recorder output. The refractive index range resulting in proper recorder output is +/-9.9x10-4RIU (with a range setting of 512). However, signal linearity is lowered when above +/- 5.0 X 10-4 RIU.

Note: When the “MEASURE RANGE” is set to WIDE, this refractive index range shifting method is invalid.

21

5. [SHIFT] Key Operations

5.1 Operation summary

The operations executed using the [SHIFT] key are summarized in Table 5.1.

Table 5.1 [Shift] Key Operations and their Functions

Key operation Function

[SHIFT][1] Preamplified output display (refer to section 5.2). Used to check for bubbles in the cell and when adjusting optical zero.

[SHIFT][3] Auto-zeroing operation (AUTO and MANUAL) (refer to section 5.3) AUTO: When a signal is input into MARK IN on the back panel, the autozeroing operation is executed automatically. MANUAL: Auto-zeroing is executed only when the [AUTO ZERO] key is pressed.

[SHIFT][4]

Zero point shift for integrator output (refer to section 5.4) The allowable input voltage for the integrator is normally within the range -10mV ~ +1V. The dynamic range on the negative side is therefore small compared to that on the positive side. When the output level is shifted beforehand to the positive side using the zero point shift function, integrator output can be prevented from falling below -10mV.

[SHIFT][5] Lamp operation time (refer to section 5.5)

[SHIFT][6] Output polarity change (refer to section 4.1.3)

[SHIFT][7] Signal filtering selection (refer to section 5.6)

[SHIFT][AUTOZERO]

Used to execute optical auto zero

[SHIFT][MARK]

Allows the following parameters to be changed.

・”MEASURE RANGE” (refer to chapter 7)

・ Constant K1 (for STD side optics calibration constant) setting (refer to section 9.4.5)

・ Constant K2 (for WIDE side optics calibration constant) setting (refer to section 9.4.5)

・ Calibration (refer to section 9.4)

・ Lamp maintenance time setting (refer to 9.5)

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・Marker voltage/polarity change (refer to section 5.7.1)

・ Setting to continue/stop self-diagnostics when an error occurs (refer to section 5.7.2)

・Serial number setting (refer to section 9.6)

5.2 Preamplified output display ([SHIFT][1])

Preamplified output is used to check for bubbles in the cell and when adjusting optical zero.


NORM 64 STD

-0.0 + ON(OFF)

[MONIT]

DIFF: -0.0200 V (*)

SAM : 1.5000 V

[SHIFT] [1][MONIT]

TEMP: xx.x/yy °C

-0.0 + ON(OFF)

Figure 5.1 Preamplified Output Screen

(1) Checking for bubbles in the cell

The existence of bubbles in the cell is evaluated by monitoring fluctuation of the SUM value. When the third digit after the decimal place is fluctuating, bubbles remain in the cell. When no fluctuation is found, no bubbles are thought to be present. However, final judgement should be made by recording and inspecting the baseline.

For example, when a problem is found with the baseline, check the SUM value. If the value has decreased, it can be assumed that bubbles have been generated. In addition, when the SUM value increases (decreases) while pumping solvent, bubbles are being eliminated (generated).

Note: The SUM value is proportional to the total light intensity detected by two photodiodes installed in the instrument. Therefore, the greatest value is restored when the cells are free of bubbles. If bubbles are

23

present, transparency is reduced and the value decreases. If the value falls below 0.1V, the INTENSITY POOR message appears.

(2) Optical zero-adjustment

When “MEASURE RANGE ” is set to STD, be sure to execute optical zero adjustment after solvent replacement. Even when using the same solvent, readjust whenever new solvent is replaced.

Note: When “MEASURE RANGE” is set to WIDE, dynamic measurement range is large. Therefore, the optical zero adjustment function is not equipped in this unit.

(3) Automatic optical zero adjustment

When “MEASURE RANGE ” is set to STD, optical zero can be automatically adjusted by pressing [SHIFT][AUTOZERO]. This key operation causes the DIFF value to fall zero within –0.01xx ~ +0.01xx. When DIFF value is not within –0.01xx ~ +0.01xx, refer to (4) below.

(4) Optical zero adjustment using [ ][ ] keys

When “MEASURE RANGE ” is set to STD, optical zero can also be adjust by [ ][ ] key operations. To increase (decrease) the DIFF value, press [ ] ([ ]) key. Adjustment is satisfactory if the value is within –0.01xx~+0.01xx. When (*) is displayed in the right side of the LCD display, the optical zero adjustment range is limited. In this case, bubbles may exist in the flow cell or replacement of solvent in the flow cell may not be enough.

Note: The value displayed for DIFF is proportional to the difference in the light intensity detected by two photodiodes. The sign of this value may be positive or negative. When the absolute value of DIFF/SUM is greater than 0.8, the instrument will cease to operate normally and the OUT OF RANGE message will appear.

5.3 Auto-zero operation setting ([SHIFT][3])

This setting determines whether or not the auto-zeroing operation will be executed automatically when a marker signal is input into the MARK IN terminal on the back panel.

24

AUTO When analyzing samples using an autosampler, the MARK IN signal, input to the detector from the autosampler at the time of sample injection, causes a marker to be overlayed on the chromatogram and auto-zero to be executed. The chromatogram base is therefore set to zero. MANUAL Auto-zeroing is not executed when a signal is input into the MARK IN terminal. The auto-zeroing operation is only executed when the [AUTO ZERO] key is pressed.

Note: The [MARK] key function on the front panel causes a marker regardless of this setting.

AUTO ZERO

AUTO

AUTO ZERO

AUTO(MANUAL)

Input screen

Flashes

[MONIT] [SHIFT] [3]

Auto-zero setting screen

[EDIT/ENTER]

(MANUAL)is the setting after the change.

[ ][ ][EDIT/ENTER]

(AUTO or MANUAL can be selected)


NORM 64 STD

-0.0 + OFF

TEMP: xx.x/yy °C

-0.0 + OFF

[MONIT]

Figure 5.2 Setting for Automatic or Manual Auto-zero Execution

Note: The AUTO setting is convenient for analysis, whereas the MANUAL setting is convenient for fractioning.

5.4 Zero point shift for integrator output ([SHIFT][4])

The allowable input voltage range of a current integrator is, in general, -10mV ~ +1V. The dynamic range on the negative side is therefore small compared to that on the positive side. When the output level is shifted beforehand to the positive side using the zero point shift function, integrator output can be prevented from falling below -10mV.

When [AUTO ZERO] is operating or when a signal is input into A/Z IN on the back panel, integrator output size of 0, 5, 10, 50, or 100mV can be selected.

25

Note 1: The zero point shift setting has no affect on recorder output.

Note 2: The extent of integrator output noise is not changed by the zero point shift setting. However, as the input voltage increases, the signal resolution of the integrator decreases and visible baseline noise increases. Therefore, do not use values that are larger than necessary

(1) Setting of integrator output zero point shift

A. Z POSITION

0 mVA. Z POSITION

0(50)mV

Input screen

Flashes

[MONIT] [SHIFT] [4]

Zero point shift setting screen

[EDIT/ENTER]

(50) is the value after the change

[ ][ ][EDIT/ENTER]


NORM 64 STD

-0.0 + OFF

TEMP: xx.x/yy °C

-0.0 + OFF

[MONIT]

Figure 5.3 Integrator Output Zero Point Shift Setting

(2) Example

When the baseline drifts to the negative during a long analysis

26

In t e g r a t o r o u t p u t

In t e g r a t o r p ro c e s s in g n o t p o s s ib le

0 V

- 1 0 m V Z e r o p o i n t s h i ft : 0 m V

0 V Z e r o p o i n t s h i ft : + 5 0 m V

In t e g r a t o r o u t p u t

Figure 5.4 Example when Baseline Drifts in Negative Direction

5.5 Lamp operation time ([SHIFT][5])

This function displays the lamp operation time.

LAMP OPERAT . TIME* 3456.7 H

[MONIT] [SHIFT] [5]

Lamp operation time screen


NORM 64 STD-0.0 + OFF

TEMP: xx.x/yy °C -0.0 + OFF

[MONIT]

Figure 5.5 Lamp Operation Time Display

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5.6 Signal filtering mode ([SHIFT][7])

The signal filtering method is normally set to “TIME ACCUME”. If a high noise level makes analysis using the “TIME ACCUME” method difficult, switch to the “DIGITAL FILTER” method. Generally, the “DIGITAL FILTER” method decreases noise, but peak shape may be broad. Select the adequate filtering method and response setting in relation to the chromatogram characteristics.

FILTER MODE TIME ACCUME

[EDIT/ENTER]

Flashes

[SHIFT] [7] [MONIT]


NORM 64 STD

-0.0 + ON(OFF)

TEMP: xx.x/yy °C

-0.0 + ON(OFF)

[MONIT]

FILTER MODE

TIME ACCUME [ ][ ][EDIT/ENTER]

(values in parenthesis are after changes)

Figure 5.6 Setting for signal filtering

Note: The digital filtering method is efficient when the full width half maximum value of the peak is wider than 5 seconds. However, when using the digital filtering method, the retention time delay becomes significant. The delay time of a peak for which the full width half maximum value is 10 seconds is described in Table 5.2.

Table 5.2 Retention time delay

Response setting Fast STD SLOW

Delay time ca. 4 sec. ca. 7 sec. ca. 13 sec.

5.7 Other settings ([SHIFT][MARK])

Pressing [SHIFT][MARK] allows the following parameters to be changed.

・”MEASURE RANGE” (refer to chapter 7)

28

・ Constant K1 (for STD side optics calibration constant) setting (refer to section 9.4.5)

・ Constant K2 (for WIDE side optics calibration constant) setting (refer to section 9.4.5)

・ Calibration (refer to section 9.4)

・ Lamp maintenance time setting (refer to section 9.5)

・Marker voltage/polarity change (refer to section 5.7.1)

・ Setting to continue/stop self-diagnostics when an error occurs (refer to section 5.7.2)

・Serial number setting (refer to section 9.6)

The calibration constants K1 and K2 must be input when the BACK UP ERROR message appears. Calibration must be performed when a light source, photodiode, or flow cell is replaced. Other items may be input as required. In this section, the marker voltage/polarity change item and the setting to continue/stop self-diagnostics when an error occurs are described. For descriptions of the other items, refer to chapter 9.

When [SHIFT][MARK] is pressed, the “MEASURE RANGE” selection menu screen appears. Press the [ ][ ] keys to scroll between the menu screens as shown in Figure 5.7.

29

PARAMETER MODE3:SET K2 VALUE

[SHIFT] [MARK][MONIT]

PARAMETER MODE2:SET K1 VALUE

[ ]


NORM 64 STD-0.0 + OFF


[MONIT]

PARAMETER MODE

1:MEASURE RANGE

PARAMETER MODE4:CALIB. K

Measurement range setting menu

[ ]

PARAMETER MODE8:SERIAL NO.

PARAMETER MODE6:MARK OUT MODE

PARAMETER MODE5:L. MAINTE. TIME

PARAMETER MODE7:DIAG. MODE

Menu to continue/stop self-diagnostics when an error occurs

Serial number menu

Constant K1 setting

Constant K2 setting

K1, K2 Calibration

Lamp maintenance time menu

Marker voltage/polarity menu

[ ] [ ]

[ ]

[ ]

[ ]

[ ]

[ ]

[ ]

[ ]

[ ]

[ ]

[ ]

Figure 5.7 Other Menus

5.7.1 Marker voltage/polarity changes

These operations are used to change the marker output voltage and polarity of Recorder output.

30

The procedures required to change the settings outlined in Figure 5.8.

Marker voltage input range: 0, 0.5, 1, 2mV

PARAMETER MODE

3:MARK OUT MODE

Marker voltage/polarity menu

Voltage input screen MARK OUT MODE

V:(1)2mV P: +(-)

MARK OUT MODE V: 1mV P: +

MARK OUT MODE V: 2mV P: +

Flashes

Polarity input screen


Marker voltage/polarity screen

[EDIT/ENTER]

[EDIT/ENTER]

Flashes

[MONIT]

[SHIFT] [MARK] [MONIT]


NORM 64 STD

-0.0 + ON(OFF) TEMP: xx.x/yy °C

-0.0 + ON(OFF)

[MONIT]

[ ] (Display the menu below by pressing the key twice)

[ ][ ][EDIT/ENTER]

Figure 5.8 Marker Voltage/Polarity Changes

5.7.2 Setting to continue/stop self-diagnostics when an error occurs

This setting is used to determine whether power on self-diagnostics will stop or continue when an error is detected and displayed. The default setting stops self-diagnostics when an error occurs. When diagnostics is set to continue and an error is detected the error message is displayed for 3 seconds then the instrument continues on to the next test.

The procedure for changing the setting to continue/stop self-diagnostics when an error occurs is outlined in Figure 5.9.

31

Input range: CONFIRM (Stop), SKIP (Continue)

[EDIT/ENTER]

DIAG MODE CONFIRM(SKIP)

[EDIT/ENTER]

Flashes

[MONIT]

PARAMETER MODE

4:DIAG. MODE



NORM 64 STD

-0.0 + ON(OFF)

TEMP: xx.x/yy °C -0.0 + ON(OFF)

[MONIT]

Menu to continue/stop self- diagnostics when an error occurs.

[ ]

DIAG MODE CONFIRM [ ][ ][EDIT/ENTER]


[ ] (Use the / keys to display the menu below)

Figure 5.9 Setting to Continue/stop Self-diagnostics when an Error Occurs

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6. Program Mode Operations

6.1 Operation summary

In program mode the range, response, valve opening and closing, and auto-zero operations can be changed according to a time program. The maximum number of steps in the time program is 64 and the maximum number of program files that can be stored is 10.

The time program operations are described in the following sections.

Switch to program mode Refer to section 6.2

Set file number Refer to section 6.3 Edit initial parameters Refer to section 6.4.1 Edit time program Refer to section 6.4.2 Execute Refer to section 6.5

6.2 Switching between normal operation mode and program mode

Press the [PRGM] key while in the normal operation mode monitor screen. The program mode monitor screen (Figure 6.1) will appear. Press the key again to return to normal mode. The program mode monitor screen consists of three screens. Switch between screens using the [MONIT] key.

33

[PRGM]

Program mode monitor screen


NORM 64 STD

-0.0 + OFF


[MONIT]

PRG2 T:0.0

-0.0 + OFF

[MONIT] TEMP: xx.x/yy °C -0.0 + OFF

[MONIT]PRG2 64 STD-0.0 + OFF

Figure 6.1 Switching between Normal Operation Mode and Program Mode

The contents of each screen are described in Figure 6.2.

Current time Program mode

(file number 2) Sensitivity

Response

Current refractive index value

Polarity Purge valve

Current heater temperature/setting temperature

Temperature control mark

PRG2 T:12.3

-0.0 + OFF

[MONIT] TEMP: xx.x/yy°C -0.0 + OFF

[MONIT]PRG2 64 STD

-0.0 + OFF

(12.3 min) (64) (standard)

blinks when heating

(positive) (off)

Figure 6.2 Meanings of Items in the Program Mode Monitor Screen

6.3 File number setting (file loading)

A maximum of 10 files (0 ~ 9) can be stored. The file number can be changed from the normal operation mode or program mode monitor screen using the following operations.

Input number range: 0 ~ 9

34

RPOGRAM FILE NO.

y

RPOGRAM FILE NO. y(x)

[SHIFT] [PRGM]

[EDIT/ENTER]

[x][EDIT/ENTER]

Flashes

File number screen Input screen

(X) is the value after the change

[MONIT]

Normal operation

mode monitor screen [PRGM] Program mode monitor

screen

Figure 6.3 File Number Change

Note 1: If an incorrect value is entered accidentally, press the [CLEAR] key before pressing the [EDIT/ENTER] key then enter the value again.

Note 2: When a file number already stored in memory is designated, the stored program will be loaded. After the file number is set and the time program is edited, the file is automatically saved. Since time programs are stored in C-MOS RAM (the memory backed up by battery), they are not erased when the power is turned off.

6.4 Program editing

6.4.1 Initial parameter editing

Range and purge valve open/close (ON/OFF) are set as initial parameters. The setting method is shown in Figure 6.4.

Note: The settings in normal operation mode for response, polarity, and temperature will remain in effect in program mode.

Note: When using a “10 ~ 50mL/min.” tubing arrangement, the solvent replacement in the reference cell is performed by drawing up the mobile phase solvent using a syringe. Solvent replacement is possible regardless of whether the purge value is ON or OFF.

35

PRG2 INITIAL

R:64(16) PURGE:OFF(ON)

[ ] [ ] [MONIT]

Initial parameter screen

[EDIT/ENTER] Purge valve

Sensitivity flashes

Purge valve flashes

Sensitivity input screen

Purge valve open/close screen

Program mode monitor screen

Sensitivity

Note: Values in parentheses are after changes

[EDIT/ENTER] [ ] [ ]

PRG2 T:0.0

-0.0 + OFF

[MONIT] TEMP: xx.x/yy°C -0.0 + OFF

[MONIT]PRG2 64 STD-0.0 + OFF

PRG2 INITIAL

R:64 PURGE:OFF

PRG2 INITIAL

R:16 PURGE: OFF


Figure 6.4 Initial Parameter Editing

6.4.2 Time program editing

6.4.2.1 Time program input

(1) Input example

As an example, a time program will be created with the initial parameters (range: 16, response: STD) held for 1 minute. The range will then be changed to 8 and, after 2 minutes, the response will be changed to FAST.

In the initial parameter screen, press the down key. The step 1 screen will appear. Follow the procedure in Figure 6.5 for subsequent operations.

36

P2 ST 2 T:0.0

P2 ST 2 T:2.0 RESPONSE STD

P2 ST 1 T:1.0

RANGE:16

P2 ST 1 T:1.0

1: PURGE ON

P2 ST 1 T:0.0

[ ] [ ]

PRG2 ST 1 T:0.0

[EDIT/ENTER]

[1][EDIT/ENTER]

[RANGE/4][EDIT/ENTER]

(waiting for input)

Input 1 minute

(waiting for input)

Call up sensitivity

(waiting for input)

Step 1 input complete

P2 ST２ T:0.0

[EDIT/ENTER]

[2][EDIT/ENTER]

[RSPNS/5][EDIT/ENTER]

(waiting for input)

Input 2 minutes

(waiting for input)

Call up response

(waiting for input)

Input FAST

Step 2 input complete

Step 1 screen Step 2 screen

[ ]▼

Initial parameter screen

: Shaded values are flashing

Time flashes

Function flashes

Value flashes

Time flashes

Function flashes

Value flashes


P2 ST 2 T:2.0 RESPONSE FAST

PRG2 INITIAL

R:16 PURGE:OFF

P2 ST 1 T:1.0

RANGE:8

Input 8

P2 ST 2 T:2.0

1: PURGE ON

or [ ] three times [EDIT/ENTER] or [ ] four times [EDIT/ENTER]


Figure 6.5 Time Program Key Operation Example

(2) Time program input method

Set the time, function, and value in sequence for each step.

a) Press the [ ] key while in the initial parameter screen (displays step 1).

b) Press [EDIT/ENTER] (the time value flashes).

c) Input the time in minutes (0.1min step).

37

[xx][EDIT/ENTER]

Function will flash next.

d) Input the function. Select a function from Table 6.1.

For example [RANGE/4][EDIT/ENTER]

The input function will appear and the value will flash.

e) Input a value.

[ ][ ][EDIT/ENTER]

When input is complete, press the [ ] key to move to the next step.

Repeat the operations in steps a) ~ e) to edit the time program. Functions that can be controlled by the time program and the relevant keys for the functions are shown in Table 6.1.

Note: Steps do not have to be input sequentially in respect to time. Even when steps are out of time sequence, they will be rearranged in chronological order once the [MONIT] key is pressed to return to the monitor screen.

Table 6.1 Function Input Keys and Input Range

Function Key Change keys Setting range Purge valve open

[P-ON/1]

Purge valve close

[P-OFF/2]

Auto-zero

[A.ZERO/3]

Range

[RANGE/4] [ ][ ] 1/4,1/2,1,2,4,8,16,32,64,128,256,512,S

Response [RSPNS/5] [ ][ ] FAST, STD, SLOW

Movement among steps

[ ]: Moves to the next step

[ ]: Moves to the previous step

38

[SHIFT][ ]: Moves to the last step in the program

[SHIFT][ ]: Moves to the first step in the program

6.4.2.2 Corrections

When time and functions do not agree

1) Use the [ ] key to display the incorrect step.

2) Re-input the step values starting from time.

When only a value, and not time and function, requires changing

1) Use the [ ] key to display the incorrect step.

2) Skip parts that do not require changes by pressing [EDIT/ENTER] until the value input screen is displayed.

3) Change the value.

For example, when changing only the range value, use the operations shown on the left in Figure 6.6. However, when input is corrected starting with time, the function selection screen, shown on the right in the figure, will appear.

39

P5 ST 3 T:1.0

1: PURGE OFF

[RANGE/4][EDIT/ENTER] Sensitivity call up

[1][EDIT/ENTER]

(waiting for input)

(waiting for input)

(Change the value here)

Time flashes

Function flashes

Value flashes

[EDIT/ENTER]

Time input

[EDIT/ENTER]

PRG2 ST 3 T:1.0

RANGE : 16

[ ][ ][EDIT/ENTER]

PRG2 ST 3 T:1.0

RANGE : 16

PRG2 ST 3 T:1.0

RANGE : 16

PRG2 ST 3 T:1.0

RANGE : 32

Figure 6.6 Correcting a Time Program

6.4.2.3 Deletion

Single step deletion

1) Display the step to be deleted using the [ ] key.

2) Set the time in the step to 0 ([EDIT/ENTER][0][EDIT/ENTER]).

3) Press the [MONIT] key to return to the monitor screen. Steps will be rearranged in chronological order and the 0 time step will be deleted.

Deletion of a step and all subsequent steps

1) Display the first step to be deleted using the [ ] key.

2) Press [SHIFT][CLEAR]. The designated step and all subsequent steps will be deleted.

40

3) Press the [MONIT] key to return to the monitor screen.

6.4.2.4 Insertion

1) Display the final step in the program using [SHIFT][ ].

2) Display a new step using the [ ] key.

3) Follow the procedure described in section 6.4.2.1 and input desired values.

4) Press the [MONIT] key to return to the monitor screen. Steps will be rearranged in chronological order.

6.5 Time program operation

6.5.1 Time program execution

Start the program while the program monitor screen is displayed.

(1) Start

After the baseline has stabilized, press the [PRGM RUN] key. The time program will start. The [PRGM RUN] key lamp will light, indicating a time program is in progress.

When the time program reaches the final step, the parameters at that point are maintained and only time advances.

(2) Stop

Press the [PRGM RUN] key during program execution. The [PRGM RUN] key lamp will go out and the time program will stop.

Note: During time program, execution parameters can be freely changed. The changed values will remain in effect until the next setting value is reached in the time program.

(3) Time program execution by external signal

The time program can be started by shorting the rear panel terminal PRGM RST/ST and GND. If the rear panel terminal PRGM RST/ST and GND are shorted while the time program is running, the time program will reset and start again immediately. When using an autosampler, the injection signal should be connected to these terminals.

41

7. MEASURE RANGE Setting

One of the major features of the RI-2031 is that the dynamic measurement range is 10 times wider than an ordinary RI detector. The “MEASURE RANGE” setting is normally set to STD, however when measuring a high concentration sample, “MEASURE RANGE” should be set to WIDE, using the procedure shown in Figure 7.1. The recorder scale and integrator output scale are automatically changed according to the “MEASURE RANGE” setting.

Table 7.1 “MEASURE RANGE” and output scale

“MEASURE RANGE” Dynamic range RANGE Inte. output

STD ~ 5x10-4RIU 1/4~512x10-6RIU 5x10-4RIU/1.0V

WIDE ~ 5x10-3RIU 1/4~512x10-5RIU 5x10-3RIU/1.0V

Note: The baseline noise and drift when set to WIDE is larger than when set to STD. Set “MEASURE RANGE” to WIDE only when the STD dynamic range makes analysis difficult.

PARAMETER SET 1:MEASURE RANGE

[SHIFT] [MARK]

[EDIT/ENTER]

[MONIT]

MEASURE RANGE STD(WIDE)

[EDIT/ENTER]

NORM 64 STD

-0.000 + ON(OFF)


[MONIT]

MEASURE RANGE STD

[MONIT]

[ ][ ][EDIT/ENTER]


(WIDE) is the setting after the change. (STD or WIDE can be selected)

Flashes

MEASURE RANGE setting screen Input screen

Figure 7.1 “MEASURE RANGE” setting

42

Note: Although signal linearity is reduced gradually when the refractive index value exceeds 5x10-4RIU (“MEASURE RANGE”: STD) or 5x10-3

RIU(“MEAUSRE RANGE”: WIDE), the measurable range is ~ 7x10-4

RIU/1.0V (“MEASURE RANGE”: STD ), and ~ 9.9x10-3RIU/1.0V (“MEASURE RANGE”: WIDE). When exceeding the linear range, connect to the recorder output terminal instead of the integrator output terminal. For example, when the RANGE setting is 128, the measurable range is 12.8x10-4 RIU/100mV (“MEASURE RANGE”: STD), and 12.8x10-3 RIU/100mV (“MEASURE RANGE”: WIDE).

43

8. Maximum Flow Rate Changing at the Front Panel

The maximum flow rate depends on the front panel tubing arrangement. The following subsections describe tubing arrangements and the method for replacing solvent in the reference cell.

8.1 “ ~ 10mL/min” tubing arrangement and solvent replacement method for reference cell

Connects ports No.1 and No.2 with outer diameter: 1.6mm, internal diameter:0.8mm tubing and ports No.4 and No.5 with outer diameter:1.6mm, internal diameter:0.8mm tubing. Connect the tubing from the column to port No.6, and connect the tubing for waste to port No.3. To replace solvent in the reference cell, operate purge valve. When replacing the reference cell solvent, set the valve to ON. During analysis, set the valve to OFF. (Refer to section 4.2)

Figure 8.1 “ ~ 10mL/min” tubing arrangement

3

4

2 1 Sample Out (waste)

Sample IN (from column)

Tubing OD=1.6mm ID= 0.8mm

Flow rate: ~ 10mL/min. 5 6

44

8.2 “10 ~ 50mL/min” tubing arrangement and solvent replacement method for reference cell

Connect the tubing from the mobile phase solvent bottle to port No3. Use tubing with an internal diameter of 0.8mm and an outer diameter of 1.6mm. Connect the syringe to port No.6. To replace solvent in the reference cell, draw up mobile phase solvent by pulling the syringe plunger. Make sure the syringe is connected during analysis. Connect the tubing from the column to port No.5, and connect the tubing for waste to port No.2. The refractive index output signal is reversed. Change the Polarity setting. (Refer to section 4.1.3)

Figure 8.2 “10 ~ 50mL/min” tubing arrangement

Tubing OD=1.6mm, ID=0.8mm

Replace solvent in the reference cell by sucking the mobile phase solvent. Keep the syringe connected during analysis in order to maintain solvent in the reference cell and to facilitate solvent replacement.

Tubing OD=1.6mm ID=0.8mm (Same tubing as used to connect ports No.1-2 or ports No.4-5. for the “~10mL/min” tubing arrangement.

Tubing OD=1.6mm ID=0.8mm

Sample

Sample IN

Reference IN

Reference Out

1

Sample Out

Sample IN

Mobile phase

2 3

4 5 6

1 2 3

4 5 6

45

9. Maintenance

9.1 Error messages and countermeasures

9.1.1 Errors during self-diagnostics after power on

The self-diagnostics program is automatically executed when the power switch is turned on. The items in Table 9.1 are checked during self-diagnostics. When a problem is found, an error message is displayed on the screen.

When an error message appears, turn the power off, wait 30 seconds, and turn the power on again. If the error message appears again, a problem exists with the instrument and the user should contact your local JASCO distributor.

Note: When the power is turned off then on quickly, a ROM CHECK ERROR or RAM CHECK ERROR may appear.

When an error message appears and self-diagnostics stop, press [SHIFT][ ] to advance to the next item and continue with the self-diagnostics program. When self-diagnostics have been advanced in this way, check that chromatograms (including baselines) are the same as previously obtained results. In particular, when BACK UP ERROR has been skipped in this way, check the following points.

(1) Have the RANGE and other setting values, including Time Program setting,

changed?

(2) Have the setting values followed [SHIFT] key operation changed?

(3) Has the K1 value changed to 1.00? (Refer to Figure 9.8 in section 9.4.5)

If setting values have changed or the K1 value has reverted to 1.00, re-input the setting values, turn the power off, wait at least 3 minutes, and turn the power on again. If the re-input values have changed again, the backup battery is dead and the user should contact your local JASCO distributor.

46

Table 9.1 Self-diagnostics Items and Corresponding Error Messages after Power On

Check item Error message

ROM operation RAM operation C-MOS RAM backup Temperature sensor Zero glass driver Lamp operation time

ROM CHECK ERROR RAM CHECK ERROR BACK UP ERROR TEMP SENSOR NG Z. GLASS ERROR WARNING LAMP MAINTE. TIME

9.1.2 Self-diagnostics during operation

The instrument checks the following items during operation (Table 9.2). When a problem is detected, an error message is displayed on the screen. Refer to section 9.1.3 for countermeasures.

Table 9.2 Self-diagnostics Items and Corresponding Error Messages

Check item Error message

Fluid leak in cell Heater overheating Light intensity deterioration Optical zero adjustment Lamp operation time

TROUBLE LEAK IN CELL TROUBLE OVER TEMP WARNING INTENSITY POOR WARNING OUT OF RANGE WARNING LAMP MAINTE TIME

9.1.3 Error message meanings and countermeasures

9.1.3.1 TROUBLE LEAK IN CELL

This message appears when fluid is leaking from the flow cell. Immediately stop the pump, turn the power off, and contact your local JASCO distributor.

47

9.1.3.2 TROUBLE OVER TEMP

This message appears when overheating occurs due to a problem with the temperature control system. The message may also appear when misjudgment happens due to a problem with the temperature detection system. Immediately turn the power off and contact your local JASCO distributor.

9.1.3.3 WARNING INTENSITY POOR

This message appears when the light intensity incident on the photodiodes is insufficient. Specifically, the message appears when the absolute value of the SUM of preamplified outputs falls below 0.1V.

When this message appears, press the [CLEAR] key to clear the message (see note), display the preamplified output screen, and check the SUM value (refer to section 5.2). When the value is smaller than 0.5V, the following problems are possible.

Note: The message will reappear every 20 seconds, until the problem is fixed.

(1) The solvent on the sample side is different from the solvent on the reference side -> Open the purge valve (ON) and pump fluid (refer to section 4.2).

(2) Large bubbles remain in the cell -> Remove the bubbles (refer to section 9.2).

(3) The cell is broken.

When the SUM value is only slightly smaller than normal, the following problems are possible. (1) Small bubbles remain in the cell -> Remove the bubbles (refer to section

9.2). (2) The cell is contaminated -> Clean the cell (refer to section 9.3).

9.1.3.4 WARNING OUT OF RANGE

This message appears when the light incident on the photodiodes is shifted to one end. Specifically, the message appears when the DIFF value of preamplified output divided by the SUM value is greater than 0.8 in absolute

48

value. The following problems are possible when the message appears.

(1) The solvent on the sample side is different from the solvent on the reference side.

(2) Large bubbles remain in the cell -> Remove the bubbles (refer to section 9.2).

(3) Optical zero is not properly adjusted. When this message appears, press the [CLEAR] key to clear the message (see note), display the preamplifed output screen, and check the DIFF value (refer to section 5.2).

Note: The message will reappear every 20 seconds, until the problem is fixed.

Replace solvent in the reference and sample cell according to the tubing arrangement. (Refer to chapter 8 and section 4.2) Once the DIFF and SUM values in the preamplified output screen have stabilized, press [SHIFT]+[AUTOZERO] key. Adjustment is satisfactory if the DIFF value falls to zero within +/-0.01xx.

9.1.3.5 WARNING LAMP MAINTE TIME

This message appears when the lamp operation time reaches the set maintenance time. Press the [CLEAR] key to clear the message. The instrument may continue to be used.

Note: Refer to section 9.5 for details regarding how to set the lamp maintenance time.

9.2 Flow cell bubble removal

9.2.1 Baselines when bubbles are generated

The following baseline phenomena occur when bubbles are generated in the flow cell.

49

(1) Irregular noise (Figure 9.1)

(2) Increased drift (Figure 9.2)

(3) Irregular ghost peaks (Figure 9.3)

When the above phenomena occur, display the preamplified output screen and check the SUM value (refer to section 5.2). If the third digit after the decimal point of the SUM value is fluctuating, it is possible that bubbles exist in the flow cell. If third digit after the decimal point is within ±1, the bubbles are considered to have been removed.

Characteristics: No periodicity, Reference: Normal noise

irregular widths

Figure 9.1 Irregular Noise Caused by Bubbles

Returns to base when bubbles are removed

Figure 9.2 Baseline Drift Caused by Bubbles

Figure 9.3 Ghost Peaks Caused by Bubbles

50

9.2.2 Bubble removal method 1 (high flow rate fluid pump)

Note: (1) When pumping fluid at a high flow rate, do not connect other detectors to the OUTLET of the RI-2031.

Note: (2) Be sure to connect a line filter to the pump outlet in order to prevent contaminants from entering the flow cell.

When using a “ ~ 10mL/min.” tubing arrangement, connect the pump directly to the RI-2031, then open and close the purge valve (ON/OFF) (refer to section 4.2) while pumping fluid at a high flow rate (up to 10mL/min is possible).

9.2.3 Bubble removal method 2 (syringe fluid injection)

Remove the INLET and OUTLET tubing of the flow cell, connect a syringe to the INLET (or OUTLET), and push solvent in (or pull solvent out).

Note: Do not move the syringe plunger too quickly. For example, if the plunger of a 20mL syringe is depressed for a full stroke in 10 seconds, the internal solvent will be flushed at a rate of 120mL/min. This greatly exceeds the maximum flow rate of the instrument and may damage the flow cell or cause fluid leaks from the electromagnetic valve.

9.2.4 Bubble removal method 3 (covering tubing outlet)

Note: Use a flow rate of 2mL/min or less with this method. If this method is used with a high flow rate, the flow cell may be damaged or fluid leaks may occur from the electromagnetic valve.

Repeatedly cover (for 2 seconds at most) and release the outlet of the OUTLET tubing. When the solvent being used is dangerous to touch, repeatedly pinch (for 2 seconds at most) and release the OUTLET tubing (Teflon tube).

9.2.5 Bubble prevention method

Follow the guidelines below (particularly when using aqueous solvents) to prevent bubble generation.

51

(1) Use a degasser.

(2) Sufficiently check for clogs in the pump inlet filter. When the inlet filter is clogged, bubbles easily enter the flow cell.

(3) Place the waste fluid bottle in a position higher than the instrument.

9.3 Flow cell cleaning method

Note1: Reversing the fluid flow direction is prohibited. Be sure to pump fluid into port No.6.

Note2: Be sure to place a line filter between the pump and the instrument.

Note3: When the inlet filter of the pump is clogged, bubbles may be introduced. This is particularly troublesome when pumping nitric acid and aqueous solutions.

When contaminants adhered to the internal cell wall are known, flush with a solvent that will dissolve the contaminants. Solvents other than the mobile phase solvent are effective.

When contaminants adhered to the internal cell wall are unknown, flush with the following solvents. Use the pump for cleaning and pump fluid at a high rate for at least 12 hours. Prepare 200mL or more of the solvents and recycle the solvents.

(1) When the mobile phase solvent is aqueous,

wash with nitric acid.

(2) When the mobile phase solvent is organic,

wash with THF (tetrahydrofurane).

52

Port No.6

Pump

RI-2031

Wash fluid

Port No.3

Figure 9.4 Flow Cell Cleaning Method

9.4 Calibration method

A sophisticated signal processing system incorporated into the RI-2031 is able to measure refractive index accurately without being affected by variations or fluctuations in lamp emission, or by the light absorption caused by a mobile phase solvent or a sample solute. (For details, refer to section 10.1.2.) However, in the following cases, recalibration is necessary because of the optical characteristics of the detector changing. This requires resetting the constant K1 and K2. The K1 value is used when “MEASURE RANGE” is set to STD and the K2 value is used when “MEASURE RANGE” is set to WIDE.

(1) After light source replacement

(2) After photodiode replacement

(3) After flow cell replacement

Note: The K1 and K2 values are recorded in RAM. When the backup battery has deteriorated, these values will be lost and the instrument will revert to the default values. The K1 and K2 values are written on a label inside the instrument. Therefore, when the values have been lost, check the K1 and K2 values on the label and re-input. (refer to section 9.4.4.)

53

9.4.1 Tubing

Figure 9.5 shows the tubing and wiring arrangement used for calibration. An injector loop with a capacity of 1mL or more is required. However, the capacity of the injector loop does not need to be precise. When an appropriate loop is not available, prepare a loop using tubing 2m or more in length with an internal diameter of 0.8mm. Alternatively, prepare a loop using tubing 1.3m or more in length with internal diameter of 1.0mm. Change to the “ ~ 10mL/min.” tubing configuration. (refer to section 8)

Pump

Water

1mL loop

Injector

Internal diameter of 0.25mm or less

Integrator or recorder

RI-2031

Port No.6

Port No.3

Figure 9.5 Tubing and Wiring Arrangement for Calibration

9.4.2 Standard solution

(1) Prepare approximately 1000mL of water. Sufficiently degas the water to prevent generation of bubbles during calibration.

(2) Dissolve 106.6mg of saccharose in a 100mL volumetric flask using the water prepared in (1) above. The refractive index of this aqueous saccharose solution is 160x10-6RIU greater than the refractive index of the water.

Note: Use the same water for the mobile phase and to dissovle the saccharose. If different water is used, the correct refractive index may not be obtained.

54

9.4.3 Replacement of tubing system solvent with water

Note: When the solvent in the cell is not miscible with water, first replace with an intermediate solvent then replace with water. For example, when the solvent in the cell is chloroform or THF (oxidation inhibitor is insoluble in water), first replace with acetone then replace with water.

(1) Pump the water prepared in section 9.4.2 (1) until the solvent in the system is sufficiently replaced.

Note: If the tubing shown in Figure 9.5 is used, since no other detectors are connected to the system, a flow rate of 10mL/min may be used

(2) Set “MEASURE RANGE” to STD. (refer to chapter 7)

(3) Wait for the drift to decrease to 1.0x10-7RIU/min or less.

Note: On the LCD display, 0.1x10-6RIU/min or less corresponds to a 0.1 RIU value per minute. When a recorder is used, this corresponds to approximately 1% or less during a 10 minutes period with a range of 64 (x10-6).

(4) Pump solvent at a flow rate of 1.0mL/min and confirm that the drift is within the limit described in (3) above. In addition, confirm that the fluctuation in the RIU value on the LCD display is less than 0.2.

9.4.4 Calibration method

(1) Following the procedure described in Figure 5.7(refer to section 5.7), display the K1 and K2 Calibration menu screen and press the [EDIT/ENTER] key. The K1 and K2 calibration screen is displayed.

55

PARAMETER SET 4:CALIB K

[SHIFT] [MARK]

[EDIT/ENTER]

[MONIT]

K CALIB: xxxx.x PRESS RUN KEY

NORM 64 STD -0.000 + ON(OFF)


[MONIT]

[MONIT]

[ ][ ][EDIT/ENTER]

K CALIB: 0.X PRESS RUN KEY

K CALIB: 160.0 PRESS RUN KEY


Once the baseline has stabilized, press the [AUTOZERO] key. Inject standard solution

Read the RIU value shown on the screen for the plateau of the peak. Press the [PRGM RUN] key to execute the built in calibration program. The RIU value will be 160±1 RIU.

Calibration screen

Figure 9.6 Calibration screen

(2) In the calibration screen, once the baseline has stabilized, press the [AUTO ZERO] key to set the RIU value to zero. Set the chart speed to 10mm/min.

(3) Inject standard solution.

(4) If the RIU value on the plateau of peak is within 160.0 ± 1.6 x 10-6 RIU, the instrument is already correctly calibrated. If the value is not within 160.0 ± 1.6 x 10-6 RIU, press the [PRGM RUN] key to execute build-in calibration program. The RIU value will automatically be set to 160.0 ± 1.6 x 10-6 RIU.

56

Plateau

Figure 9.7 Peak Shape

Note: When the peak dose not contain a plateau, inspect the internal

diameter of the tubing from the injector to the detector. When the plateau is short, decrease the flow rate.

(5) Repeat the operations in steps (2) ~ (4) and confirm that the RIU value is within 160.0 ± 1.6 x 10-6 RIU without pressing the [PRGM RUN] key.

Note: Adjustment of the integrator or recorder output is not necessary. The outputs will automatically be adjusted when the K1 and K2 values are calibrated.

Note: Calibration of constants K1 and K2 is performed by executing build-in calibration program at the same time.

9.4.5 Calibration constant confirmation and setting

After performing calibration (outlined in previous section), the calibration constants, K1 and K2 obtained should be recorded. These values should be written on a piece of paper and attached inside the instrument. This way, the recorded values can then be re-input in the following situations, and troublesome calibration operations can be avoided. When confirming and setting the calibration constant, follow the procedure shown in Figure 9.8 and re-input the calibration constants.

(1) When the K1 or K2 values have been changed by mistake

57

(2) When the backup battery has deteriorated and the K1 value has reverted to 1.00.

(3) When the PCB (CPU board) has been replaced

Note: At the time of shipping, the K1 and K2 values are recorded on a label attached inside the instrument. This label can be seen when the case is removed.

PARAMETER SET 2:SET K1 VALUE

[SHIFT] [MARK]

[EDIT/ENTER]

[MONIT]

K1 VALUE: x.xxx RIU: xxxx.x

NORM 64 STD -0.000 + ON(OFF)

TEMP: xx.x/yy°C -0.000 + ON(OFF)

[MONIT]

[MONIT]

[ ][ ]

PARAMETER SET 3:SET K2 VALUE


[EDIT/ENTER] [MONIT]

[ ][ ]

[EDIT/ENTER]


[MONIT]


[EDIT/ENTER] [MONIT]


Figure 9.8 Confirming and setting calibration constant

9.4.6 Alternative calibration method

The method described in sections 9.4.1 ~ 9.4.4 is the most accurate method of calibration.

However, since the entire flow line, including the pump, must be replaced with

58

water, the operations are troublesome when non-aqueous mobile phases (chloroform for example) are being used. It may be possible to replace the solution in the detector using only a syringe.

A detailed description of the procedure will not be given, however, points of caution when using syringe injection are listed below.

(1) Do not move the syringe plunger too quickly. For example, if the plunger of a 20mL syringe is depressed for a full stroke in 10 seconds, the internal solvent will be flushed at a rate of 120mL/min. This greatly exceeds the maximum flow rate of the instrument and may damage the flow cell or cause fluid leaks from the electromagnetic valve.

(2) When a syringe is used to inject fluid, there is a tendency to introduce bubbles into the flow cell. Although this will not cause a problem during solvent replacement, proper calibration is not possible if bubbles are introduced immediately before or after standard solution injection. Display the preamplified output on the screen and confirm that no bubbles are present while performing calibration (refer to section 5.2).

9.5 Lamp maintenance time setting

Lamp maintenance time can be set to inform the operator of the number of hours the lamp has been in use. When the lamp operation time exceeds the lamp maintenance time, the LAMP MAINTENANCE warning will be displayed. Since lamp replacement is done by service engineer, contact your local JASCO distributor when this is required.

When the lamp maintenance time is set to 0, the warning is not displayed. In addition, when BACKUP ERROR is displayed due to deterioration of the battery, the lamp maintenance time recorded in RAM will be lost. When this occurs, follow the procedure in Figure 9.9 and re-input the lamp maintenance time.

59

[EDIT/ENTER]

[xxxxxx] [EDIT/ENTER]

LAMP MAINTE TIMER 10000 H

[EDIT/ENTER]

Flashes

[MONIT]

PARAMETER MODE

2:L. MAINTE. TIME



NORM 64 STD

-0.0 + ON(OFF) TEMP: xx.x/yy °C -0.0 + ON(OFF)

[MONIT]

LAMP MAINTE TIMER 10000 H

Lamp maintenance time menu

[ ] (Use the / keys to display the menu below)[ ]

Figure 9.9 Lamp Maintenance Time Input Method

9.6 Serial number setting

The serial number of the instrument is recorded on the label on the right side of the instrument case. When BACKUP ERROR is displayed due to deterioration of the battery, the serial number recorded in RAM will be lost. When this occurs, use the procedure outlined in Figure 9.10 to re-input the serial number. When connecting to a JASCO system controller via LC-Net, the serial number is displayed on the HPLC system configuration screen.

60

PARAMETER MODE

6:SERIAL NO.

[EDIT/ENTER]

[xxxxxxxxxx] [EDIT/ENTER]

SERIAL NO 1234567890

[EDIT/ENTER]

Flashes

[MONIT]

SERIAL NO 1234567890



NORM 64 STD

-0.0 + OFF

TEMP: xx.x/yy°C

-0.0 + OFF

[MONIT]

Serial number menu

[ ] (Use the / keys to display the menu below)[ ]

Figure 9.10 Serial Number Input Method

9.7 Power Fuse replacement

Warning: When changing the power fuses, be sure to disconnect the power cable from the power cable inlet (refer to Figure 2.5).

Warning: For continued protection against risk of fire, replace only with fuses of the specified type and current ratings.

Note: When one fuse is blown, replace both fuses.

61

Insert finger under the fuse holder, pinch the holder, and pull forward. The fuses and the fuse holder will come out together. (Figure 9.11)

(1) Pull the old fuses out from fuse holder, insert new fuses, and insert the fuse holder back into its original position.

If the replaced fuses immediately blow out again, an instrument problem is possible. Contact your local JASCO distributor.

Fuse Holder

Figure 9.11 Fuse Holder

62

10. Appendix

10.1 Operation Theory

10.1.1 Operation theory of deflection differential refractometers

With deflection differential refractometers, light refracted inside the prism flow cell is used to detect the difference in refractive indices between the reference (solvent) and the sample.

The flow cell structure and light path are shown in Figure 10.1.

i

n

n

S

R

i

θ

r

Figure 10.1 Horizontal Cross-section of Flow Cell and Light Path

The area to the left of the center partition is called the sample cell and the area to the right is called the reference cell.

ns : Refractive index of sample

nR : Refractive index of reference (solvent)

θ: Angle of deflection (exaggerated in the figure: actual value is <10-2 degree.)

i: Angle of partition (determined by cell shape)

Applying the law of refraction to this light path and assuming θ << 1 (since the actual value is 10-2 or less), θ can be expressed as follows:

θ = tan i x(nS-nR)/nR... (1)

From this equation, the angle of deflection is proportional to the difference in refractive indices between the reference solvent and the sample. In other words, it is possible to determine the difference in refractive indices between the reference solvent and sample by measuring the angle of deflection.

63

10.1.2 Optical system

Light source Slit

First photodiode

Collimator lens

Flow cell Half mirror

Second photodiode

Zero glass (motor driven)

Figure 10.2 Optical System

Light from the light source passes through a slit and into the collimator lens where it is made into a parallel beam. The parallel light then passes through and is refracted by the flow cell. Part of the light is reflected by a half mirror, passes through the cell again, and strikes the first photodiode. The remainder of the light passes through the half mirror and strikes the second photodiode.

If the concentration of the sample in the cell changes, then the refractive index changes, thereby changing the angle of deflection in accordance with equation (1). The light image striking the primary photodiode will therefore shift in the horizontal plane (direction perpendicular to the plane of this paper).

The amount of shift is proportional to the angle of deflection and the focal length of the lens. When s is the amount of shift and f is the focal length of the lens, the following equation can be expressed.

s = 2f x θ... (2)

The coefficient of 2 is required in the equation to account for the fact that the light passes through the flow cell twice and is therefore refracted twice.

The method for detecting the amount of shift is described below.

The first and second photodiodes contain identical photo-sensitive surfaces and are referred to as PD1 and PD2, respectively.

64

PD1 output PD2 output

PD1 output PD2 output

No difference between refractive indices

Difference between refractive indices

Figure 10.3 Light Image on Photodiodes

First, when there is no difference between the refractive indices of the reference (solvent) and the sample, the light image overlaps PD1 and PD2 equally as shown in the top half of Figure 10.3. Since the electrical output from a photodiode is proportional to the surface area receiving light, the difference between the output of PD1 and the output of PD2 is zero and so DIFF is zero.

Next, when there is a difference between the refractive indices, the image is shifted, as shown in the bottom half of Figure 10.3, and a difference between the output of PD1 and the output of PD2 is generated. Since the output is proportional to the surface area receiving light, the output difference is proportional to the amount of shift s.

DIFF = A x s...(3)

In this equation, A is a constant determined by the light source intensity, photodiode measurable range, flow cell transmittance, and amplification of the amplifier. Equations (1) ~ (3) can be combined to give the following.

nS-nR = B x DIFF...(4)

where B = 1/2f tan i x (nR/A)

In other words, the difference between the refractive indices of the reference solvent and sample is proportional to DIFF, that is the difference between the outputs from PD1 and PD2. Therefore, the difference between the refractive indices can be calculated by measuring the difference between the outputs of PD1 and PD2.

65

In the case of an optical system with a second photodiode, the distance between the flow cell and second photodiode is shorter than that of an optical system with first photodiode. This means that for the optical arrangement shown in Figure 10.2, 2f in equation (2) is 10 times smaller and, therefore, the dynamic range is 10 times larger.

10.1.3 Signal processing method

Theoretically, the difference between refractive indices can be found by measuring DIFF, as shown in equation (4). Although A in equation (4) is proportional to light source intensity and flow cell transmittance, and both of these factors can change over time.

On the other hand, the sum of the outputs from PD1 and PD2 (hereafter called SUM) is proportional to the light source intensity and flow cell transmittance regardless of the value of the refractive index.

SUM = A x k...(5)

The following can then be obtained using equations (4) and (5).

nS - nR - K x DIFF/SUM

where K = knR/(2f tan i)

DIFF/SUM is proportional only to the difference between refractive indices and does not change over time. Therefore, the value is not affected by light source deterioration or flow cell contamination.

The RI-2031 measurement system uses two calibration constants (K1 and K2). K1 value is for analytical measurement (first photodiode) and K2 value is for preparative measurement (second photodiode).

66

10.1.4 Flow line

A flow line diagram for the “ ~ 10mL/min.” tubing arrangement, is shown in Figure 10.4. Effluent form the column is introduced from the port No.6 and waste is discharged from the port No.3. Flow lines are changed using the purge valve.

When the purge valve is ON, the instrument is switched to the flow line indicated by the dotted arrow in Figure 10.4. In this case, the effluent passes through the sample cell and then through the reference cell before being discharged.

When the purge valve is OFF, the instrument is switched to the flow line indicated by the solid arrow in Figure 10.4. In this case, the effluent passes through the sample cell only before being discharged and the reference cell is completely sealed.

Figure 10.4 Flow Line (“ ~ 10mL/min.” tubing arrangement)

6

3

1

4

5

2

OUT

IN

Sam.

Ref.

During analysis (PURGE OFF)

When replacing solvent in the reference cell (PURGE ON)

Heat exchanger

Flow cell

Purge

valve

Purge OFF

Purge ON

Heat exchanger

67

A flow line diagram for the “10 ~ 50mL/min.” tubing arrangement, is shown in Figure 10.5. Effluent from the column is introduced from the port No.5 (Sample IN) and waste is discharged from the port No.2 (Sample OUT). The mobile phase solvent is introduced from the port No.3 (Reference IN) and is drawn using the syringe, from the port No.6 (Reference OUT). The sample and reference flow lines are independent.

The sample side flow line is indicated by the solid arrow in Figure 10.5. The solvent from the column passes through the heat exchanger and then through the sample cell.

The reference side flow line is indicated by the dotted arrow in Figure 10.5. When replacing the solvent in the reference cell, draw up the mobile phase solvent using a syringe connected to the No.6 port. The mobile phase passes through the reference cell and then through heat exchanger.

Note: The tubing connection between ports No.1 and No.4 avoids introducing air into the reference cell.

Figure 10.5 Flow Line (“10 ~ 50mL/min.” tubing arrangement)

Sample side flow line

Reference side flow line

6

3

1

4

5

2

Reference IN

Reference OUT

Ref.

Sam.

Heat exchanger

Flow cell

Purge

valve

Purge OFF

Purge ON

Heat exchanger

Sample IN

Sample OUT

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10.1.5 Internal volume of tubing

Internal volume of tubing is described in Table 10.1 and Table 10.2.

Table 10.1 Internal volume using “ ~ 10mL/min.” tubing

Flow line Volume

Port No6 -> Sample cell ca. 70 µL

Sample cell -> Port No3 ca. 300µL

Table 10.2 Internal volume using “10 ~ 50mL/min.” tubing

Flow line Volume

Port No5 -> Sample cell ca. 500 µL

Sample cell -> Port No2 ca. 200 µL

Operation Manual RI 2031aa

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