Jasco RI-2031 Instruction Manual

MODEL RI-2031

INTELLIGENT REFRACTIVE INDEX

DETECTOR

INSTRUCTION MANUAL

P/N: 0302-0555A

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.

i

(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

ii

o

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.

Warning

This is a Class A product. In a domestic environment this product may cause radi

interference, in which case the user may be required to take adequate measures.

• 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 equipment generates, uses, and can radiate radio frequency energy. If it is not

installed and used in accordance with the instruction manual, it may cause

interference 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.

iii

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.

iv

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.

v

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:

y Copying of software or related materials for purposes other than

backup is prohibited.

y Transfer or authorization of the use of Jasco software to or by a third

party is prohibited.

y Disclosure of confidential information related to Jasco software is

prohibited.

y Modification of Jasco software is prohibited.

y 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.

vi

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

vii

viii ix

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

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

xi

xii

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.

1

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

5.0 x 10

-4

RIU (“MEASURE RANGE”: STD)

-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> H

2

O

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)

2

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.

3

2. Part Names and Functions

2.1 Front panel

Operation Panel

Solvent Inlet and Outlet

POWER Switch



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.

4

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.)

Tubing with

OD=1.6mm

ID= 0.8mm

1

4

5

2

3

6

Sample Out (waste)

Flow rate: ~ 10mL/min.

Sample IN (from column)

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

5

g

6

6

Tubing

OD=1.6mm

ID=0.8mm

(Same tubing as used

to connect ports

No.1-2 or ports

Sample Out

1

2

4 5

Reference IN

3

Flow rate: 10 ~ 50mL/min.

No.4-5. for the “

10mL/min” tubing

arran

ement.

Tubing

OD=1.6mm

ID=0.8mm

~

Sample Out

1

4

2 3

5

Sample IN

Reference Out

Tubing OD=1.6mm, ID=0.8mm

Mobile phase

Flow rate: 10 ~ 50mL/min.

Sample IN

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.

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

6

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.

7

2.3 Back panel

Input/Output Terminals

LC-Net Connectors

RS-232C Terminal

Fuses

AC input

Ground Terminal

Figure 2.5 Back Panel

8

Name Function
Input and output terminals
Signal output terminals
REC+/­G(GND)­INT+/­G(GND)

MARK OUT +/-

LEAK OUT +/-

Signal input terminals
MARK IN+
GND­A/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).

9

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.

10



Mode display
(Normal operation mode)

MODE RANGE RESPONSE

Sensitivity

(64)

NORM 64 STD

-0.1 + ON

RIU(x10 ) POLARITY PURGE

Current
refractive index
(-0.1)

-6

Polarity
(Positive)

Normal operation mode monitor screen(1)

Figure 3.1 Normal Operation Mode Screens

3.2 Power off

Response

(standard)

[MONIT]

Purge valve status
(ON)

Current Optical bench
temperature/setting

temperature

MODE RANGE RESPONSE

Temperature

TEMP: xx.x/yy 

-0.1 + ON

RIU(x10 ) POLARITY PURGE

-6

°C



control mark



blinks
when heating

Normal operation mode monitor screen(2)

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.

11





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.

Mode display
(Normal operation mode)

MODE RANGE RESPONSE

NORM 64 STD

-0.1 + ON

RIU(x10 ) POLARITY PURGE

Current
refractive index
(-0.1)

-6

Polarity

(Positive)

Sensitivity

(64)

Purge valve status
(ON)

Response
(standard)

[MONIT]

Current optical bench

temperature/setting
temperature

MODE RANGE RESPONSE

TEMP: xx.x/yy 

-0.1 + ON

RIU(x10 ) POLARITY PURGE

-6

°C




Temperature
control mark

blinks
when heating

Normal operation mode monitor screen(1)

Normal operation mode monitor screen(2)

Figure 4.1 Normal Operation Mode Screens

12

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

NORM 64(16) STD(FAST)

-0.000 + ON

[MONIT]

NORM 64 STD

-0.000 + ON

NORM 16 STD

-0.000 + ON

[EDIT/ENTER]



[ ][ ][EDIT/ENTER]





[ ][ ][EDIT/ENTER]



Normal operation mode monitor screen(1)

Values in parentheses are after changes.

Flashes

Sensitivity input screen

Display the desired value

Flashes

Response rate input screen

Display the desired value

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.

13

NORM 64(16) STD

-0.000 + ON

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

NORM 64 STD

-0.000 + ON

Sensitivity change





Sensitivity flashes

Figure 4.3 Change Method (2)

[RSPNS/5]

NORM 64 STD(FAST)

-0.000 + ON

[ ][ ][EDIT/ENTER]





NORM 64 STD

-0.000 + ON

Response change

Response flashes

14

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.

TEMP: xx.x/yy(zz) °C 

-0.0 + ON

[EDIT/ENTER][MONIT]

TEMP: xx.x/yy °C 

-0.0 + ON

Note:Temperature control will be turned off



Normal operation mode monitor screen(2)



Flashes




Temperature input screen

when 0 is input

Values in parentheses are after changes.

Values input[zz][EDIT/ENTER]

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).

15

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

RANGE”: STD) or 5x10

-4

RIU (“MEASURE RANGE”: WIDE), integrator output

-5

RIU (“MEASURE

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

STD

WIDE

WIDE

+ -5.0×10

- -5.0×10
+ -5.0×10

- -5.0×10

-6

~+5.0×10-4 RIU -5.0×10-5~+4.5×10

-4

~+5.0×10

-5

~+5.0×10

-3

~+5.0×10

-6

RIU -4.5×10

-3

RIU -5.0×10

-5

RIU -4.5×10

-4

~+5.0×10

-4

~+4.5×10

-3

~+5.0×10

-4

RIU

-5

RIU

-3

RIU

-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

STD

WIDE

WIDE

- -5.0×10
+ -5.0×10

- -5.0×10
+ -5.0×10

-6

~+5.0×10-4 RIU -5.0×10-5~+4.5×10

-4

~+5.0×10-6 RIU -4.5×10

-5

~+5.0×10

-3

~+5.0×10

-3

RIU -5.0×10

-5

RIU -4.5×10

-4

~+5.0×10

-4

~+4.5×10

-3

~+5.0×10

-4

RIU

-5

RIU

-3

RIU

-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.

16

＋

0

−

Actual refractive index change

RIU:10.0

0

RIU:-20.0

Polarity:Positive

RIU:20.0

0

RIU:-10.0

Polarity:Negative

Figure 4.5 Changes in Peak Appearance and Display Values Caused by

Polarity Differences

NORM 64 STD

-0.0 + ON

[MONIT]

POLARITY

+(-)

(-)is the value after the change.

Output polarity change screen

[SHIFT] [5]

Normal operation mode monitor screen(1)

[EDIT/ENTER]

[ ][ ][EDIT/ENTER]





POLARITY

+

Flashes

Input screen

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.

17





Normal operation mode monitor screen

NORM 64 STD

-0.0 + ON(OFF)

[MONIT]

[MONIT]

PURGE ON

1:ON 0:OFF

[1]

:Opens the purge valve and changes the status to ON.

[0]

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

TEMP: xx.x/yy °C

-0.0 + ON(OFF)

[PURGE]

Current purge valve status

Purge valve open/close screen




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

18

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

4.5 Measurement of concentrated sample (Refractive index range shifting

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

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

19

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.

20

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]

[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][AUTO
ZERO]

[SHIFT][MARK] Allows the following parameters to be changed.

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.

Used to execute optical auto zero

・”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)

21









・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.

Normal operation mode monitor screen

NORM 64 STD

-0.0 + ON(OFF)

[MONIT]

[MONIT]

TEMP: xx.x/yy °C

-0.0 + ON(OFF)

[SHIFT] [1]




DIFF: -0.0200 V (*)

SAM : 1.5000 V

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

22

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.

23





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.

Normal operation mode monitor screen

NORM 64 STD

-0.0 + OFF

[MONIT]

AUTO ZERO

AUTO(MANUAL)

(MANUAL)is the setting after the change.

Auto-zero setting screen

[SHIFT] [3]

[MONIT]

[EDIT/ENTER]

[ ][ ][EDIT/ENTER]





TEMP: xx.x/yy °C

-0.0 + OFF

(AUTO or MANUAL can be selected)




AUTO ZERO

AUTO

Flashes

Input screen

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.

24





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

Normal operation mode monitor screen

NORM 64 STD

-0.0 + OFF

[MONIT]

A. Z POSITION

[SHIFT] [4]

0(50)mV

(50) is the value after the change

Zero point shift setting screen

[MONIT]

TEMP: xx.x/yy °C

-0.0 + OFF

[EDIT/ENTER]

[ ][ ][EDIT/ENTER]








A. Z POSITION

0 mV

Flashes

Input screen

Figure 5.3 Integrator Output Zero Point Shift Setting

(2) Example

When the baseline drifts to the negative during a long analysis

25





Integrator o utput

0 V

-10 m V

Integrator o utput

Zero point shift:0m V

Integrator p roc ess ing n ot possible

0 V

Figure 5.4 Example when Baseline Drifts in Negative Direction

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

This function displays the lamp operation time.

Normal operation mode monitor screen

NORM 64 STD

-0.0 + OFF

[MONIT]

LAMP OPERAT . TIME

[MONIT]

[SHIFT] [5]

* 3456.7 H

Zero point shift:+5 0m V

TEMP: xx.x/yy °C

-0.0 + OFF




Lamp operation time screen

Figure 5.5 Lamp Operation Time Display

26





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.

Normal operation mode monitor screen

NORM 64 STD

-0.0 + ON(OFF)

[MONIT]

FILTER MODE

TIME ACCUME

(values in parenthesis are after changes)

[SHIFT] [7]

[MONIT]

[EDIT/ENTER]

[ ][ ][EDIT/ENTER]



TEMP: xx.x/yy °C

-0.0 + ON(OFF)






FILTER MODE

TIME ACCU ME

Flashes

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)

27

・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.

28





Normal operation mode monitor screen

NORM 64 STD

-0.0 + OFF

PARAMETER MODE

1:MEASURE RANGE

[ ]



PARAMETER MODE

2:SET K1 VALUE

[ ]  [ ]

PARAMETER MODE

3:SET K2 VALUE

[ ]



PARAMETER MODE

4:CALIB. K

[ ] 

[SHIFT] [MARK][MONIT]

[ ]





[ ]



[ ]



[MONIT]

Measurement range setting menu

Constant K1 setting

Constant K2 setting

K1, K2 Calibration

TEMP: xx.x/yy °C

-0.0 + OFF




PARAMETER MODE

5:L. MAINTE. TIME

[ ] 

PARAMETER M ODE

6:MARK OUT MODE

[ ]



PARAMETER MODE

7:DIAG. MODE

[ ]



PARAMETER MODE

8:SERIAL NO.

[ ]



[ ]



[ ]



Lamp maintenance time menu

Marker voltage/polarity menu

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

Serial number 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.

29





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

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

Normal operation mode monitor screen

NORM 64 STD

-0.0 + ON(OFF)

[MONIT]

TEMP: xx.x/yy °C

-0.0 + ON(OFF)

[SHIFT] [MARK] [MONIT]

(Display the menu below by pressing the key twice)

[ ]



Marker voltage/polarity menu

PARAMETER MODE

3:MARK OUT MODE

[MONIT]

MARK OU T MO DE

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

[EDIT/ENTER]

[EDIT/ENTER]

Marker voltage/polarity screen

(values in parenthesis are

after changes)

MARK OUT MODE

V: 1mV P: +

Flashes

MARK OUT MODE

V: 2mV P: +




Voltage input screen

[ ][ ][EDIT/ENTER]





Polarity input screen

Flashes

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.

30





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

Normal operation mode monitor screen

NORM 64 STD

[MONIT]

-0.0 + ON(OFF)

[MONIT]

(Use the

[ ]



[SHIFT] [MARK]

[ ]

keys to display the menu below)



/

Menu to continue/stop self­diagnostics when an error occurs.

PARAMETER MODE

4:DIAG. MODE

[MONIT]

DIAG MODE

CONFIRM(SKIP)

[EDIT/ENTER]

[EDIT/ENTER]

[ ][ ][EDIT/ENTER]





(values in parenthesis are after changes)

TEMP: xx.x/yy °C

-0.0 + ON(OFF)




DIAG MODE
CONFIRM

Flashes

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

31

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.

32











Normal operation mode monitor screen

NORM 64 STD

-0.0 + OFF

PRG2 T:0.0

-0.0 + OFF

[MONIT]

[MONIT]

[PRGM]

PRG2 64 STD

-0.0 + OFF

TEMP: xx.x/yy °C

-0.0 + OFF

[MONIT]




TEMP: xx.x/yy °C

-0.0 + OFF

Program mode monitor screen

Figure 6.1 Switching between Normal Operation Mode and Program Mode

The contents of each screen are described in Figure 6.2.

Current heater temperature/setting temperature

Temperature control mark

[MONIT]

TEMP: xx.x/yy°C



-0.0 + OFF

(off)

Program mode
(file number 2)

PRG2 T:12.3

-0.0 + OFF

Current time

(12.3 min)

[MONIT]

Current refractive
index value

Sensitivity

(64)

Response

(standard)

PRG2 64 STD

-0.0 + OFF

Polarity
(positive)

Purge valve




blinks when heating




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

33

Normal operation
mode monitor screen

[MONIT]

RPOGRAM FILE NO.

y(x)

(X) is the value after the change

File number screen

[SHIFT] [PRGM]

[PRGM]

[x][EDIT/ENTER]

[EDIT/ENTER]

Program mode monitor
screen

RPOGRAM FILE NO.

y

Flashes

Input 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.

34





Program mode monitor screen

PRG2 T:0.0

-0.0 + OFF

[ ]



PRG2 INITIAL

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

Sensitivity

[EDIT/ENTER]

PRG2 INITIAL

R:64 PURGE:OFF

Sensitivity

flashes

[EDIT/ENTER]



PRG2 INITIAL

R:16 PURGE: OFF

[MONIT]

[ ]



Purge valve

[ ]



PRG2 64 STD

[MONIT]

-0.0 + OFF

[MONIT]

Initial parameter screen

Note: Values in parentheses are

after changes

Sensitivity input screen

[ ]

Purge valve open/close screen

Purge valve flashes

TEMP: xx.x/yy°C

-0.0 + OFF




[EDIT/ENTER] [ ]  [ ]



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.

35











PRG2 INITIAL

R:16 PURGE:OFF

[ ]



[ ]



PRG2 ST 1 T:0.0

[EDIT/ENTER]

P2 ST 1 T:0.0

[1][EDIT/ENTER]

P2 ST 1 T:1.0

1: PURGE ON

Call up sensitivity

[RANGE/4][EDIT/ENTER]

three times [EDIT/ENTER]

or [ ]



P2 ST 1 T:1.0

RANGE:16

Initial parameter screen

Step 1 screen

Time flashes

(waiting for input)

Input 1 minute

Function flashes

(waiting for input)

Value flashes

(waiting for input)

[ ]

▼

P2 ST２ T:0.0



[EDIT/ENTER]

P2 ST 2 T:0.0

[2][EDIT/ENTER]

P2 ST 2 T:2.0

1: PURGE ON

Call up response
[RSPNS/5][EDIT/ENTER]

[ ]

or

four times [EDIT/ENTER]



P2 ST 2 T:2.0

RESPONSE STD



Step 2 screen

Tim e flas h e s

(waiting for input)

Input 2 minutes

Function flashes

(waiting for input)

Value flashes

(waiting for input)

Inp ut 8

[ ]

[EDIT/ENTER]





P2 ST 1 T:1.0

RANGE:8

[ ]

Step 1 input

complete

[ ][ ]



[EDIT/ENTER]

P2 ST 2 T:2.0

RESPONSE FAST

Input FAST

Step 2 input
complete



: Shaded values are flashing

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).

36

[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

Range

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

[A.ZERO/3]

[RANGE/4] [][] 1/4,1/2,1,2,4,8,16,32,64,

128,256,512,S

Movement among steps

[]: Moves to the next step

[]: Moves to the previous step

37

[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.

38

PRG2 ST 3 T:1.0

 RANGE : 16 

[EDIT/ENTER]

PRG2 ST 3 T:1.0

 RANGE : 16 

[EDIT/ENTE R]

PRG2 ST 3 T:1.0

 RANGE : 16

[ ][ ][EDIT/ENTER]





PRG2 ST 3 T:1.0

 RANGE : 32 

Figure 6.6 Correcting a Time Program

Tim e fl as h es

(waiting for input)

[1][EDIT/ENTER]

P5 ST 3 T:1.0

1: PURGE OFF

[RANGE/4][EDIT/ENTER]

Value flashes

(Change the value here)

Time inp u t

Function flashes

(waiting for input)

Sensitivity call up

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.

39

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.

40













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.

Normal operation mode monitor screen

NORM 64 STD

-0.000 + ON(OFF)

[MONIT]

PARA METE R SET 

1:MEASURE RANGE

[MONIT]

MEASURE RANGE



[EDIT/ENTER]

STD(WIDE)

[MONIT]

[SHIFT] [MARK]

[EDIT/ENTE R]

[ ][ ][EDIT/ENTER]





TEMP: xx.x/yy °C

-0.000 + ON(OFF)

MEASURE RANGE




STD

Flashes

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

MEASURE RANGE setting screen

Input screen



Figure 7.1 “MEASURE RANGE” setting

41

Note: Although signal linearity is reduced gradually when the refractive index

value exceeds 5x10-4RIU (“MEASURE RANGE”: STD) or 5x10

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

-3

-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

STD), and 12.8x10

-3

RIU/100mV (“MEASURE RANGE”: WIDE).

-4

RIU/100mV (“MEASURE RANGE”:

42

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)

1

2

3

Sample Out (waste)

Tubing

OD=1.6mm

4

Flow rate: ~ 10mL/min.

ID= 0.8mm

Sample IN (from column)

Figure 8.1 “ ~ 10mL/min” tubing arrangement

43

6

6

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)

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.

Sample Out

1

Tubing

OD=1.6mm

ID=0.8mm

4

2

5

Sample

1

4

Sample IN

Tubing OD=1.6mm, ID=0.8mm

Reference IN

2

3

5

Reference Out

Mobile phase

Sample IN

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.

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

44

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.

45

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

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

ROM CHECK ERROR

RAM CHECK ERROR

BACK UP ERROR

TEMP SENSOR NG

Z. GLASS ERROR

WARNING LAMP MAINTE.
TIME

Check item Error message

Fluid leak in cell

Heater overheating

Light intensity deterioration

Optical zero adjustment

Lamp operation 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.

TROUBLE LEAK IN CELL

TROUBLE OVER TEMP

WARNING INTENSITY POOR

WARNING OUT OF RANGE

WARNING LAMP MAINTE TIME

46

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

47

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.

48

(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

49

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.

50

(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).

51

6

Port No.3

RI-2031

Pump

Wash fluid

Port No.

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.)

52

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)

Integrator or

recorder

Port No.3

1mL loop

RI-2031

Pump

Injector

Internal diameter of 0.25mm or less

Port No.6

Water

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

-6

saccharose solution is 160x10

RIU 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.

53

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

-7

RIU/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.

54























Normal operation mode monitor screen

NORM 64 STD

-0.000 + ON(OFF)

[MONIT]

PAR A METE R S E T 
4:CALIB K

[MONIT]

K CALIB: xxxx.x

PRESS RUN KEY

K CALIB: 0.X

PRESS RUN KEY

[MONIT]

[SHIFT] [MARK]
[ ][ ][EDIT/ENTER]

 



[EDIT/ENTER]

Calibration screen

Once the baseline has stabilized, press

the [AUTOZERO] key.

Inject standard solution

TEMP: xx.x/yy °C

-0.000 + ON(OFF)




Read the RIU value shown on the screen

K CALIB: 160.0

PRESS RUN KEY

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.



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 ±

-6

1.6 x 10

program. The RIU value will automatically be set to 160.0 ± 1.6 x 10

RIU, press the [PRGM RUN] key to execute build-in calibration

-6

RIU.

55



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

-6

within 160.0 ± 1.6 x 10

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

RIU without pressing the [PRGM RUN] key.

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

56















 





(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.

Normal operation mode monitor screen

NORM 64 STD

-0.000 + ON(OFF)

[MONIT]

PAR A METE R S E T 

2:SET K1 VALUE

[MONIT]

K1 VALUE: x.xxx

RIU: xxxx.x

[MONIT]

K1 VALUE: x.xxx

RIU: xxxx.x

[MONIT]

[SHIFT] [MARK]

[ ][ ]







[ ][ ]



[EDIT/ENTER]

[EDIT/ENTER]

TEMP: xx.x/yy°C 

-0.000 + ON(OFF)

PARAMETER SET

3:SET K2 VALUE

[EDIT/ENTER] [MONIT]

K2 VALUE: x.xxx

RIU: xxxx.x

[MONIT]

[EDIT/ENTER]

K2 VALUE: x.xxx

RIU: xxxx.x






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

57

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.

58





Normal operation mode monitor screen

NORM 64 STD

[MONIT]

-0.0 + ON(OFF)

[ ]

[SHIFT] [MARK]



[MONIT]

[ ]

(Use the / keys to display the menu below)



TEMP: xx.x/yy °C 

-0.0 + ON(OFF)

Lamp maintenance time menu

PARAMETER MODE

2:L. MAINTE. TIME

[MONIT]

LAMP MAINTE TIMER

10000 H

[EDIT/ENTER]

[EDIT/ENTER]

[xxxxxx] [EDIT/ENTER]




LAMP MAINTE TIMER

10000 H

Flashes

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.

59



Normal operation mode monitor screen

NORM 64 STD

-0.0 + OFF

[ ]

[ ]

(Use the /





Serial number menu

PARAMETER MODE

6:SERIAL NO.

[MONIT]

SERIAL NO

1234567890

Figure 9.10 Serial Number Input Method

[MONIT]

[SHIFT] [MARK] [MONIT]

keys to display

[EDIT/ENTE R]

[xxxxxxxxxx] [EDIT/ENTER]

TEMP: xx.x/yy°C 

-0.0 + OFF

the menu below)

[EDIT/ENTE R]




SERIAL NO

1234567890

Flashes

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.

60

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

61

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.

n

S

i

i

r

n

R

θ

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.

n

: Refractive index of sample

s

n

: Refractive index of reference (solvent)

R

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

-2

actual value is 10

or less), θ can be expressed as follows:

θ = tan i x(n

S-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.

62













10.1.2 Optical system

Collimator lens

Flow cell

Second photodiode

Half mirror

Light source

Slit

First 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.

63

PD1 output

No difference between refractive indices

PD1 output

Difference between refractive indices

PD2 output

PD2 output

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.

= B x DIFF...(4)

n

S-nR

where B = 1/2f tan i x (n

R

/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.

64

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).

- nR - K x DIFF/SUM

n

S

where K = kn

/(2f tan i)

R

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).

65

r

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.

Heat exchanger

Sam.

Ref.

Flow cell

Heat exchange

6

Purge OFF

3

Purge

valve

Purge ON

1

4

5

2

During analysis (PURGE OFF)

When replacing solvent in the reference cell (PURGE ON)

IN

OUT

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

66

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.

Heat exchanger

Purge OFF

Ref.

Flow cell

Sam.

Heat exchanger

Purge

valve

Purge ON

6

Reference OUT

3

Reference IN

1

4

5

Sample IN

2

Sample OUT

Sample side flow line

Reference side flow line

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

67

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

68