Operation Manual RI 2031aa
March 7, 2017 | Author: athanasiosm | Category: N/A
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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.
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(2) Warning Labels Warning labels are attached at several locations on this instrument. Do not remove, deface or damage the warning labels. If a warning label peels off the instrument or becomes illegible, contact your local JASCO distributor for a replacement label. Be sure to indicate the part number (P/N) on the label. (1) Fuse and Ground Warning Label
Figure 1
(P/N: 0822-0163A)
Figure of the side view
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Regulatory Statements CE Notice Marking by the symbol indicates compliance of this JASCO system to the EMC (Electromagnetic Compatibility) and Low Voltage Directives of the European Community. This symbol indicates that this JASCO system meets the relevant basic safety and health requirements of the EC Directive based on the following technical standards: • EN55011 -- ”Limits and Methods of Measurement of Radio Interference Characteristics of Information Technology Equipment.” -- Group 1, Class A. Warning This is a Class A product. In a domestic environment this product may cause radio 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 Statement
Commission
Radio
Frequency
Interference
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.
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Preface This instruction manual serves as a guidebook for using this instrument. It is intended to instruct first-time users on how to properly use the instrument, and to serve as a reference for experienced users. Before using the instrument, please read this instruction manual carefully, and make sure that the contents are fully understood. This manual should be easily accessible to the operator at all times during instrument operation. When not using the instrument, keep this manual stored in a safe place. Should this instruction manual be lost, order a replacement from your local JASCO distributor.
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Installation Conditions To ensure safe operation, the following recommendations should be observed: (1)
Do not operate the instrument under voltage fluctuations exceeding 10% of the recommended line voltage. Large fluctuations may cause the instrument to fail.
(2)
Use a three-pronged electrical outlet with a ground. When only a two-pronged socket is available, use an adapter and be sure to connect the ground wire of the adapter.
(3)
Operate the instrument under a temperature range of 10 ∼ 30°C.
(4)
Operate the instrument under a humidity range of 35 ∼ 85% (RH). If ambient humidity exceeds 85% (RH), water vapor may deteriorate optical components. If possible, install the instrument in a location having a humidity of 60% or lower.
(5)
Operate the instrument under an atmospheric pressure of 750 ∼ 1060 hPa.
(6)
Avoid strong magnetic fields and sources of high-frequency waves. The instrument may not function properly when near strong magnetic fields or high-frequency wave sources.
(7)
Avoid vibrations caused by vacuum pumps, electric motors, processing equipment and machine tools.
(8)
Avoid dust and corrosive gas. Do not install the instrument in a location where it may be exposed to dust, especially in locations exposed to outside air or ventilation outlets that discharge dust particles.
(9)
Do not install the instrument in a location where it may be exposed to direct sunlight.
(10)
Do not install the instrument in a location where it may be directly exposed to the air current from an air conditioner or heater, as such a location may inhibit stable measurement.
Note:
The above conditions do not ensure optimal performance of this instrument. 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.
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Warranty This product is warranted for a period of one year from the date of delivery. If any defects should occur in the product during this period of warranty, JASCO will repair or replace the defective part(s) or product free of charge. This warranty does not apply to defects as a result of the following: (1) USE FOLLOWING IMPROPER OR INADEQUATE INSTALLATION. (2) IMPROPER OPERATION. (3) MOVEMENT, MODIFICATION, OR REPAIR BY PERSONS OTHER THAN AUTHORIZED JASCO PERSONNEL. (4) USE OF PARTS OTHER THAN THOSE THAT ARE AUTHORIZED BY JASCO. (5) INORDINATELY RAPID DETERIORATION DUE TO THE USE OF CORROSIVE SOLVENTS OR SAMPLES. (6) NATURAL DISASTERS SUCH AS FIRES, WATER DAMAGE, OR EARTHQUAKES. In addition, this warranty does not cover: (7) CONSUMABLE PARTS OR PARTS THAT HAVE A SEPARATE WARRANTY OR A WARRANTY PERIOD THAT IS DIFFERENT THAN THAT SPECIFIED ABOVE. The warranty period for all parts and repairs supplied under this warranty expires with the warranty period of the original product.
Jasco Corporation
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Table of Contents Safety Considerations ...................................................................i Regulatory Statements ................................................................iii Preface..........................................................................................iv Installation Conditions .................................................................v Maintenance .................................................................................vi Warranty ......................................................................................vii Table of Contents.........................................................................ix 1.
Outline and Specifications...................................................1
1.1 Outline .................................................................................................... 1 1.2 Specifications ........................................................................................ 2
2.
Part Names and Functions ..................................................4
2.1 Front panel ............................................................................................. 4 2.2 Operation panel ..................................................................................... 7 2.3 Back panel.............................................................................................. 8
3.
Power On/Off and Self-diagnostics...................................10
3.1 Power on and self-diagnostics ........................................................... 10 3.2 Power off .............................................................................................. 11
4.
Operations in Normal Operation Mode .............................12
4.1 Parameter changes.............................................................................. 12 4.1.1 Range and response changes..................................................... 12 4.1.2 Temperature control ON/OFF and temperature setting ............ 15 4.1.3 Output polarity change method ([SHIFT][6]) .............................. 15 4.2 Purge valve open/close (Solvent replacement in the reference cell) ............................................................................................................. 17 4.3 Auto-zero operations........................................................................... 18 4.4 Error and warning messages during monitoring.............................. 18
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4.5 Measurement of concentrated sample (Refractive index range shifting................................................................................................. 19
5.
[SHIFT] Key Operations .....................................................21
5.1 Operation summary............................................................................. 21 5.2 Preamplified output display ([SHIFT][1]) ........................................... 22 5.3 Auto-zero operation setting ([SHIFT][3])............................................ 23 5.4 Zero point shift for integrator output ([SHIFT][4])............................. 24 5.5 Lamp operation time ([SHIFT][5]) ....................................................... 26 5.6 Signal filtering mode ([SHIFT][7]) ....................................................... 27 5.7 Other settings ([SHIFT][MARK]) ......................................................... 27 5.7.1 Marker voltage/polarity changes................................................. 29 5.7.2 Setting to continue/stop self-diagnostics when an error occurs ................................................................................................................ 30
6.
Program Mode Operations.................................................32
6.1 Operation summary............................................................................. 32 6.2 Switching between normal operation mode and program mode .... 32 6.3 File number setting (file loading) ....................................................... 33 6.4 Program editing ................................................................................... 34 6.4.1 Initial parameter editing ............................................................... 34 6.4.2 Time program editing................................................................... 35 6.5 Time program operation...................................................................... 40 6.5.1 Time program execution.............................................................. 40
7.
MEASURE RANGE Setting.................................................41
8.
Maximum Flow Rate Changing at the Front Panel...........43
8.1 “ ~ 10mL/min” tubing arrangement and solvent replacement method for reference cell................................................................................. 43 8.2 “10 ~ 50mL/min” tubing arrangement and solvent replacement method for reference cell................................................................... 44
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9.
Maintenance........................................................................45
9.1 Error messages and countermeasures.............................................. 45 9.1.1 Errors during self-diagnostics after power on........................... 45 9.1.2 Self-diagnostics during operation .............................................. 46 9.1.3 Error message meanings and countermeasures....................... 46 9.2 Flow cell bubble removal .................................................................... 48 9.2.1 Baselines when bubbles are generated ..................................... 48 9.2.2 Bubble removal method 1 (high flow rate fluid pump).............. 50 9.2.3 Bubble removal method 2 (syringe fluid injection) ................... 50 9.2.4 Bubble removal method 3 (covering tubing outlet)................... 50 9.2.5 Bubble prevention method .......................................................... 50 9.3 Flow cell cleaning method .................................................................. 51 9.4 Calibration method .............................................................................. 52 9.4.1 Tubing ........................................................................................... 53 9.4.2 Standard solution ......................................................................... 53 9.4.3 Replacement of tubing system solvent with water.................... 54 9.4.4 Calibration method....................................................................... 54 9.4.5 Calibration constant confirmation and setting .......................... 56 9.4.6 Alternative calibration method .................................................... 57 9.5 Lamp maintenance time setting ......................................................... 58 9.6 Serial number setting .......................................................................... 59 9.7 Power Fuse replacement..................................................................... 60
10. Appendix .............................................................................62 10.1 Operation Theory ............................................................................... 62 10.1.1 Operation theory of deflection differential refractometers ..... 62 10.1.2 Optical system ............................................................................ 63 10.1.3 Signal processing method......................................................... 65 10.1.4 Flow line ...................................................................................... 66 10.1.5 Internal volume of tubing........................................................... 68
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1.
Outline and Specifications
1.1 Outline The JASCO RI-2031 Intelligent Refractive Index Detector is an HPLC detector which offers unprecedented versatility, performance, and stability. The RI-2031 features the following: High stability High stability is obtained through an optical system that is not easily influenced by temperature, an efficient thermal exchanger, and a digital signal processing system. Wide dynamic range 10 times larger dynamic range than an ordinary HPLC refractive index detector. Maximum flow rate of 50mL/min. Maximum flow rate is 50mL/min. (Solvent: H2O) Wide dynamic range and Extended Maximum flow rate are applied to the semi-preparation. Easy operation Parameters can be easily set using the LCD display, function keys, and numeric keys. Self-diagnostics results and warning messages are also displayed, which allows the condition of the instrument to be accurately monitored. Optical Zero can be performed precisely using the easy key operation. Automatic analysis capability The internal time program allows automatic switching of sensitivity range, response and other functions. Using LC-Net, parameters can be set from the Model HSS-2000 System without an interface. Compact design The front panel dimensions are the same as other JASCO 2000 series HPLC equipment (pump, UV detector), which facilitates a neat stacking arrangement. Safety measures An internal fluid leak sensor is installed. When a leak from the flow cell occurs, a signal is output and the pump can be stopped.
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1.2 Specifications Model name:
RI-2031 Intelligent Refractive Index Detector
Measurement system:
Deflection type
Refractive index range:
1.00 ~ 1.75
Measurement range settings:
1/4, 1/2, 1, 2, 4, 8, 16, 32, 64, 128, 256, 512 x 10-6RIU/full scale (“MEASURE RANGE”: STD) 1/4, 1/2, 1, 2, 4, 8, 16, 32, 64, 128, 256, 512 x 10-5RIU/full
scale
(“MEASURE
RANGE”:
WIDE) Linearity:
5.0 x 10-4 RIU (“MEASURE RANGE”: STD) 5.0 x 10-3 RIU (“MEASURE RANGE”: WIDE) H2O
Noise:
0.2 x 10-8RIU H2O, Flow stopped
Cell capacity:
10µL
Maximum usable flow rate:
10mL/min. (“ ~ 10mL/min.” tubing) 50mL/min. (“10 ~ 50mL/min.” tubing) H2O
Maximum pressure:
0.1MPa. (“ ~ 10mL/min.” tubing) 0.3MPa. (“10 ~ 50mL/min.” tubing)
Temperature control:
PID control by heater, Setting range: 0 ~ 45°C in 1°C increments. Control range: Room temperature +10°C ~ Room temperature +25°C
Zero adjustment:
Automatic Optical and electronic auto-zero functions. Integrator output zero positioning
Reference selection:
Electromagnetic valve control (“ ~ 10mL/min.” tubing) 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: Marker, auto-zero, program circuit each LC-Net control: Liquid wetted material:
1 circuit each reset/start: 1 1 circuit
Quartz, fluoride resin, stainless steel (SUS316)
Dimensions and weight:
150(W) x 470(D) x 150(H) mm, excluding Protrusions. Approximately 13kg
Power requirements:
AC100 ~ 240V, AC, 50/60Hz, 50VA max.
Ambient temperature:
+10 ~ +35°C for operation -30 ~ +60°C for storage
* Specifications are subject to change without notice.
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2.
Part Names and Functions
2.1 Front panel
Operation Panel
Solvent Inlet and Outlet
POWER Switch
Figure 2.1 Front Panel Name Solvent Inlet Outlet “1” ~ “6” Operation panel POWER switch Note:
Function and 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
Suitable tubing for the Solvent Inlet and Outlet connections must be selected according to maximum flow rate. Before connecting tubing, refer to Figure 2.2. and Figure 2.3. “ ~ 10mL/min”. tubing is supplied with the instrument.
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Note: For a “10 ~ 50mL/min” tubing arrangement, the refractive index signal output (and display) polarity becomes negative. Change the polarity setting from plus (+) to minus (-) using the polarity setting function. (Refer to 4.1.3 polarity setting.)
Tubing
with
1
2
3
4
5
6
Sample Out (waste)
Flow rate: ~ 10mL/min.
OD=1.6mm ID= 0.8mm Sample IN (from column)
Figure 2.2 “ ~ 10mL/min” flow rate tubing connections
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Sample Out
Reference IN
Tubing OD=1.6mm ID=0.8mm
1
2
3
4
5
6
(Same tubing as used to connect ports No.1-2 or ports
Flow rate: 10 ~ 50mL/min.
No.4-5. for the “ ~ 10mL/min” tubing arrangement.
Sample IN
Reference Out
Tubing OD=1.6mm, ID=0.8mm Sample Out
1
2
3
4
5
6
Tubing OD=1.6mm
Mobile phase
ID=0.8mm Flow rate: Sample IN
10 ~ 50mL/min.
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
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2.2 Operation panel
Figure 2.4 Operation Panel Name LCD screen [PRGM RUN] [AUTO ZERO] [MARK] [MONIT] [SHIFT] [PURGE] [PRGM] [0] ~ [9], [.]
[ ][ ] keys [CLEAR] [EDIT/ENTER]
Function Displays operation conditions, setting values, error messages and other information. 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. Sets the refractive index unit value (RIU) to zero. Outputs a marker signal from the recorder output terminal. Used to return to the monitor screen from other screens and to switch between monitor screens. Used in combination with other keys to set parameters that are not frequently changed. Executes solvent replacement (ON/OFF) for the reference cell. Switches between normal and time program mode. 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. Used to edit parameters and change screens. Used to erase incorrect parameter input and clear error messages. Used to edit parameters.
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2.3 Back panel
Input/Output Terminals
LC-Net Connectors
RS-232C Terminal
Fuses AC input
Ground Terminal
Figure 2.5 Back Panel
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Name 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+ GNDPRGM RST/ST+ GNDCooling fan AC input Power fuses Ground terminal LC-Net connector RS-232C terminal
Function Recorder output terminal (Output level determined by RANGE setting) Integrator output terminal (Fixed scale of 5x10-4RIU/1V(“MEASURE RANGE”: STD), 5x10-3RIU/1V(“MEASURE RANGE”: WIDE). Marker output terminal (Normally open, contact type output is closed when [MARK] key is pressed or MARK IN is input) Fluid leak warning output terminal (Normally open, contact type output is closed when a fluid leak from the flow cell occurs) (Note) The internal fluid leak sensor can only detect a leak of buffer solution. Marker input terminal (Marker is added on the recorder output when the contact point is closed) Auto-zero input terminal (Recorder and integrator outputs are zeroed when contact type input is closed) Time program input terminal (Time program is reset and immediately started when contact type input is closed) Used for cooling the internal components of the instrument Accepts the power cable. Protects the instrument from excessive currents. Grounds the instrument. This is not the ground terminal for input and output signals. Input and output terminal for the LC-Net. Used when controlling the instrument via a RS-232C cable (optional).
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3.
Power On/Off and Self-diagnostics
3.1 Power on and self-diagnostics Turn the power switch, located at the lower left of the front panel, to the ON position. The self-diagnostics program is automatically executed. The program examines the following items: ROM CHECK (Memory) RAM CHECK (Memory) DC POWER (Direct current power source) BACK UP (Memory backed up by battery) TEMP SENSOR (Temperature sensor) LAMP CHECK (Lamp energy) LAMP O.P TIME (Lamp operating time) ZERO GLASS DRV. (Optical zero glass driver) An error message will appear if a problem is detected. Note:
When an error occurs, diagnostics will either stop immediately or continue to the next item depending on the instrument settings (refer to section 5.7.2).
The screen on the left in Figure 3.1 will appear when diagnostic tests are complete. Press the [MONIT] key to switch to the screen on the right in Figure 3.1. The heater temperature is displayed in the screen on the right. When the [MONIT] key is pressed again, the display will return to the screen on the left. Normal operation mode consists of these two screens.
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Response (standard)
Sensitivity Mode display (Normal operation mode) (64) MODE
RANGE
NORM -0.1
64 +
MODE
RESPONSE
STD
[MONIT]
-6
Polarity (Positive)
RANGE
RESPONSE
TEMP: xx.x/yy °C -0.1
ON
RIU(x10
RIU(x10 ) POLARITY PURGE
Current refractive index (-0.1)
Current Optical bench temperature/setting temperature
-6
+
ON
Temperature control mark blinks when heating
) POLARITY PURGE
Purge valve status (ON)
Normal operation mode monitor screen(1)
Normal operation mode monitor screen(2)
Figure 3.1 Normal Operation Mode Screens
3.2 Power off Turn the power switch, located at the lower left of the front panel, to the OFF position. The range, response, polarity, temperature, and time program set at this time will be recorded in C-MOS RAM (memory backed up by battery). These values will be restored when the power is turned on again.
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4.
Operations in Normal Operation Mode
The dynamic measurement range of the RI-2031 is 10 times larger than ordinary HPLC refractive index detectors. The dynamic measurement range of the RI-2031 depends on the “MEASURE RANGE” setting. When a highly concentrated sample is analyzed, set the “MEASURE RANGE” to WIDE. Refer to section 7. The maximum flow rate of the RI-2031 depends on the tubing connections of the front cell panel. The “ ~ 10mL/min.” tubing connections are for conventional analysis and the “10 ~ 50mL/min” tubing connections are for semi-preparative analysis. Tubing connection configurations are described in section 8.
4.1 Parameter changes 4.1.1 Range and response changes After the power is turned on and the self-diagnostic tests are complete, the normal operation mode monitor screen (1) (left in Figure 4.1) will appear. Range and response settings can be changed here. Note:
Use the [MONIT] key to switch between monitor screens.
Response (standard)
Mode display Sensitivity (Normal operation mode) (64) MODE
RANGE
NORM -0.1
64 +
RESPONSE
STD
MODE
[MONIT]
RESPONSE
TEMP: xx.x/yy °C
RIU(x10
RIU(x10 ) POLARITY PURGE
Polarity (Positive)
RANGE
-0.1
ON
-6
Current refractive index (-0.1)
Current optical bench temperature/setting temperature
-6
+
ON
Temperature control mark blinks when heating
) POLARITY PURGE
Purge valve status (ON)
Normal operation mode monitor screen(1)
Normal operation mode monitor screen(2)
Figure 4.1 Normal Operation Mode Screens
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Input range: 1/4, 1/2, 1, 2, 4, 8, 16, 32, 64, 128, 256, 512, S (Short) x10-6RIU /10mV (“MEASURE RANGE”: STD) 1/4, 1/2, 1, 2, 4, 8, 16, 32, 64, 128, 256, 512, S (Short) x10-5RIU /10mV (“MEASURE RANGE”: WIDE) Response input range: FAST, STD, SLOW (1) Change method 1 NORM 64(16) STD(FAST) Normal operation mode monitor screen(1) Values in parentheses are after changes. -0.000 + ON [MONIT] NORM -0.000
NORM -0.000
[EDIT/ENTER] 64
STD
+
ON
Flashes Sensitivity input screen
[
][EDIT/ENTER] Display the desired value Flashes STD Response rate input screen ON
[
][
16 +
][
][EDIT/ENTER] 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.
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NORM
64(16) STD
NORM
64 STD(FAST)
-0.000
+
-0.000
+
ON
[RANGE/4]
[
][
][EDIT/ENTER]
ON [
[RSPNS/5]
][
Sensitivity flashes NORM -0.000
64
STD
+
ON
Response flashes NORM -0.000
Sensitivity change
64
STD
+
ON
Response change
Figure 4.3 Change Method (2)
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][EDIT/ENTER]
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
+
[MONIT]
ON [EDIT/ENTER]
TEMP: xx.x/yy °C -0.0
Normal operation mode monitor screen(2) Values in parentheses are after changes.
+
ON
Flashes Temperature input screen
[zz][EDIT/ENTER] Values input Note:Temperature control will be turned off when 0 is input
Figure 4.4 Temperature Control ON/OFF and Temperature Setting
4.1.3 Output polarity change method ([SHIFT][6]) This operation changes the output polarity of the signal. When “10 ~ 50mL/min.” tubing is used, or negative peak heights exceed the integrator input range, polarity should be set to negative (-). 4.1.3.1 “10 ~ 50mL/min.” tubing arrangement When “10 ~ 50mL/min.” tubing is used, because of the optical arrangement, the refractive index signal output decreases when the actual refractive index increases. Therefore, the polarity should be set to negative (-) in order to correct the relation between the actual refractive index and the refractive index signal output (for both the recorder and the integrator). 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 5x10-5 RIU (“MEASURE RANGE”: STD) or 5x10-4 RIU (“MEASURE RANGE”: WIDE), integrator output can be prevented from falling below –10mV using the zero point shift function. Refer to section 5.4. When the negative peak signal exceeds this range, polarity should be changed. The actual allowable refractive index range is shown in Table 4.1 and Table 4.2. Change polarity settings, according to these tables.
Table 4.1 “ ~ 10mL/min.” tubing arrangement (*shifted by 100mV) MEASURE RANGE polarity STD
+
STD
-
WIDE
+
WIDE
-
without zero point shift* -6
-4
-4
-6
-5
-3
-3
-5
-5.0×10 ~+5.0×10
-5.0×10 ~+5.0×10 -5.0×10 ~+5.0×10 -5.0×10 ~+5.0×10
RIU RIU RIU RIU
with zero point shift* -5
-4
RIU
-4
-5
RIU
-4
-3
RIU
-3
-4
RIU
-5.0×10 ~+4.5×10
-4.5×10 ~+5.0×10 -5.0×10 ~+4.5×10 -4.5×10 ~+5.0×10
Table 4.2 “10 ~ 50mL/min.” tubing arrangement (*shifted by 100mV) MEASURE RANGE polarity STD STD WIDE WIDE
+ +
without zero point shift* -6
-4
-4
-6
-5
-3
-3
-5
-5.0×10 ~+5.0×10
-5.0×10 ~+5.0×10 -5.0×10 ~+5.0×10 -5.0×10 ~+5.0×10
RIU RIU RIU RIU
with zero point shift* -5
-4
RIU
-4
-5
RIU
-4
-3
RIU
-3
-4
RIU
-5.0×10 ~+4.5×10
-4.5×10 ~+5.0×10 -5.0×10 ~+4.5×10 -4.5×10 ~+5.0×10
When polarity is set to negative (-) with a “ ~ 10mL/min” tubing arrangement, the screen display and output (for both the recorder and the integrator) will change as shown in Figure 4.5. When the actual refractive index decreases, the index (RIU) value on the screen will increase and so will output. Changes in peak appearance and display values caused by polarity differences are shown in Figure 4.5 and key operations are shown in Figure 4.6.
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RIU:20.0
＋ RIU:10.0 0
0
0 RIU:-10.0
−
RIU:-20.0
Actual refractive index change
Polarity:Positive
Polarity:Negative
Figure 4.5 Changes in Peak Appearance and Display Values Caused by Polarity Differences NORM -0.0
64
STD
+
ON
Normal operation mode monitor screen(1)
[SHIFT] [5]
[MONIT]
POLARITY
[EDIT/ENTER]
+(-)
POLARITY +
[ ][ ][EDIT/ENTER] (-)is the value after the change.
Flashes
Output polarity change screen
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 -0.0
64
STD
[MONIT]
TEMP: xx.x/yy °C -0.0 + ON(OFF)
+ ON(OFF) [MONIT] PURGE 1:ON
[PURGE] ON 0:OFF
Current purge valve status Purge valve open/close screen
[1] :Opens the purge valve and changes the status to ON. [0] :Closes the purge valve and changes the status to OFF.
Figure 4.7 Purge Valve Open/Close Operations Note:
When using “10 ~ 50mL/min” tubing connections, draw up the mobile phase solvent using a syringe in order to replace solvent in the reference cell. Replacement of solvent in the reference cell can be performed regardless of the PURGE VALVE setting.
4.3 Auto-zero operations When the [AUTO ZERO] key is pressed, the current refractive index value is set as 0. In other words, the recorder and integrator output are set to 0. Note:
When a signal is input into MARK IN on the back panel, the auto-zero operation can be set to occur automatically. (Refer to section 5.3)
Note:
When executing optical auto zero, press the [SHIFT][AUTOZERO] key. (Refer to section 5.2)
4.4 Error and warning messages during monitoring The error and warning messages that may occur during monitoring are shown in Table 4.3. When an error message is displayed, press the [CLEAR] key to return to the 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 Fluid is leaking from the flow cell TROUBLE OVER TEMP Heater is overheating TROUBLE ZERO GLASS DRV Zero glass drive not move WARNING INTENSITY POOR Light intensity has deteriorated WARNING OUT OF RANGE Optical zero adjustment is not correct WARNING LAMP MAINT TIME Lamp maintenance time has been exceeded
4.5 Measurement of concentrated sample (Refractive index range shifting when the “MEASURE RANGE” setting is STD) When “MEASURE RANGE” is set to STD, the linear dynamic range of refractive index, with properly adjusted optical zero, is +/-5.0x10-4RIU. Although this range cannot be expanded, it can be shifted in one direction. For example, the allowable refractive index range can be shifted to -3.0x10-4 ~ +7.0x10-4RIU. The method for shifting the range is described below. (1) Display the preamplified output screen (refer to section 5.2) and press the [ ][ ] keys so that DIFF is within 0±0.03 (2) Return to the monitor screen. (3) Press the [AUTO ZERO] key to set the RIU value to 0. (4) Display the preamplified output screen and press the [ ][ ] keys until the RIU value reaches -300.0±30(x10-6RIU) (this assumes the output polarity is set to positive). (5) Again press the [AUTO ZERO] key to set the RIU value to 0. As long as optical zero adjustment is not executed, the allowable refractive index range will be -3.0x10-4 ~ 7.0x10-4RIU. Note:
The refractive index range resulting in proper integrator output is +/-5.0x10-4RIU, taking into consideration the polarity setting. When this 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]
Zero point shift for integrator output (refer to section 5.4) The allowable input voltage for the integrator is normally within the range -10mV ~ +1V. The dynamic range on the negative side is therefore small compared to that on the positive side. When the output level is shifted beforehand to the positive side using the zero point shift function, integrator output can be prevented from falling below -10mV.
[SHIFT][5]
Lamp operation time (refer to section 5.5)
[SHIFT][6]
Output polarity change (refer to section 4.1.3)
[SHIFT][7]
Signal filtering selection (refer to section 5.6)
[SHIFT][AUTO ZERO]
Used to execute optical auto zero
[SHIFT][MARK]
Allows the following parameters to be changed. ・”MEASURE RANGE” (refer to chapter 7) ・ Constant K1 (for STD side optics calibration constant) setting (refer to section 9.4.5) ・ Constant K2 (for WIDE side optics calibration constant) setting (refer to section 9.4.5) ・ Calibration (refer to section 9.4) ・ Lamp maintenance time setting (refer to 9.5)
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 -0.0
64
[MONIT]
STD
+ ON(OFF)
TEMP: xx.x/yy °C -0.0
+ ON(OFF)
[SHIFT] [1]
[MONIT]
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 -0.0 [MONIT]
64 +
STD
[MONIT]
TEMP: xx.x/yy °C
OFF
-0.0
+
OFF
[SHIFT] [3]
AUTO ZERO AUTO(MANUAL)
[EDIT/ENTER] [
][
AUTO ZERO
][EDIT/ENTER]
(MANUAL)is the setting after the change. Auto-zero setting screen
AUTO
Flashes (AUTO or MANUAL can be selected) 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 -0.0
[MONIT]
64 +
[MONIT]
STD
OFF
TEMP: xx.x/yy °C -0.0
+
OFF
[SHIFT] [4]
A. Z POSITION 0(50)mV
[EDIT/ENTER] [
][
][EDIT/ENTER]
A. Z POSITION 0 mV Flashes
(50) is the value after the change
Input screen
Zero point shift setting screen
Figure 5.3 Integrator Output Zero Point Shift Setting (2) Example When the baseline drifts to the negative during a long analysis
25
In t e g r a t o r o u t p u t
0 V Z e r o p o i n t s h i ft : 0 m V
-1 0 m V
In t e g r a t o r p r o c e s s i n g n o t p o s s i b l e
In t e g r a t o r o u t p u t
0 V
Z e r o p o i n t s h i ft : + 5 0 m 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 -0.0
64 +
[MONIT]
STD
TEMP: xx.x/yy °C -0.0
OFF
+
OFF
[SHIFT] [5]
[MONIT]
LAMP OPERAT . TIME * 3456.7 H 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 -0.0
64
[MONIT]
STD
TEMP: xx.x/yy °C
+ ON(OFF)
[MONIT]
-0.0
+ ON(OFF)
[SHIFT] [7] [EDIT/ENTER]
FILTER MODE TIME ACCUME
[
][
FILTER MODE
][EDIT/ENTER]
TIME ACCUME
(values in parenthesis are after changes)
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 -0.0
64 +
STD
[MONIT]
OFF
[MONIT]
TEMP: xx.x/yy °C -0.0
+
OFF
[SHIFT] [MARK]
PARAMETER MODE
Measurement range setting menu
1:MEASURE RANGE [
]
[
]
PARAMETER MODE 2:SET K1 VALUE [
]
[
]
PARAMETER MODE 3:SET K2 VALUE [
[
]
]
[
Constant K2 setting
]
PARAMETER MODE 4:CALIB. K [
Constant K1 setting
K1, K2 Calibration
]
PARAMETER MODE
Lamp maintenance time menu
5:L. MAINTE. TIME [
]
[
]
PARAMETER MODE
Marker voltage/polarity menu
6:MARK OUT MODE [
]
[
]
PARAMETER MODE 7:DIAG. MODE [
]
[
Menu to continue/stop self-diagnostics when an error occurs
]
PARAMETER MODE 8:SERIAL NO.
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]
[MONIT]
TEMP: xx.x/yy °C -0.0 + ON(OFF)
[SHIFT] [MARK]
(Display the menu below by pressing the [
] key twice)
Marker voltage/polarity menu PARAMETER MODE 3:MARK OUT MODE [MONIT]
[EDIT/ENTER]
MARK OUT MODE
MARK OUT MODE
[EDIT/ENTER]
V: 1mV
V:(1)2mV P: +(-) Marker voltage/polarity screen (values in parenthesis are after changes)
Flashes
P: + [
][
][EDIT/ENTER]
MARK OUT MODE V: 2mV
Voltage input screen
P: +
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 -0.0 + ON(OFF)
[MONIT] (Use the [
[MONIT]
TEMP: xx.x/yy °C -0.0
+ ON(OFF)
[SHIFT] [MARK] ]/ [
] keys to display the menu below)
Menu to continue/stop selfdiagnostics when an error occurs. PARAMETER MODE 4:DIAG. MODE [EDIT/ENTER]
[MONIT]
[EDIT/ENTER]
DIAG MODE CONFIRM(SKIP)
[
][
][EDIT/ENTER]
(values in parenthesis are after changes)
Figure 5.9
DIAG MODE CONFIRM Flashes
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 [MONIT]
NORM 64 STD -0.0 + OFF
TEMP: xx.x/yy °C -0.0 + OFF
[PRGM] PRG2 T:0.0 -0.0 + OFF
[MONIT]
PRG2 64 STD -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. Program mode (file number 2) PRG2 -0.0
T:12.3 +
Current time (12.3 min) [MONIT]
Sensitivity (64) PRG2 -0.0
OFF Current refractive index value
Response (standard)
64 +
STD
Current heater temperature/setting temperature
[MONIT]
OFF
Polarity (positive)
Temperature control mark blinks when heating TEMP: xx.x/yy°C -0.0 + OFF
Purge valve (off)
Figure 6.2 Meanings of Items in the Program Mode Monitor Screen
6.3 File number setting (file loading) A maximum of 10 files (0 ~ 9) can be stored. The file number can be changed from the normal operation mode or program mode monitor screen using the following operations. Input number range: 0 ~ 9
33
Normal operation mode monitor screen
[MONIT]
[PRGM]
Program mode monitor screen
[SHIFT] [PRGM]
[EDIT/ENTER] RPOGRAM FILE NO. RPOGRAM FILE NO. y(x) y [x][EDIT/ENTER] (X) is the value after the change
Flashes
File number screen
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 [MONIT]
PRG2 T:0.0 -0.0 + OFF [
-0.0
]
PRG2
PRG2
[
64 +
]
STD
[MONIT]
OFF
TEMP: xx.x/yy°C -0.0 + OFF
[MONIT]
INITIAL
Initial parameter screen Note: Values in parentheses are after changes Purge valve [EDIT/ENTER]
R:64(16) PURGE:OFF(ON) Sensitivity PRG2
INITIAL
R:64 PURGE:OFF Sensitivity flashes PRG2
[
][
] [EDIT/ENTER]
INITIAL
R:16 PURGE: OFF [
][
Sensitivity input screen
Purge valve open/close screen Purge valve flashes
] [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
Initial parameter screen
R:16 PURGE:OFF [
]
[
PRG2 ST 1
]
[ ▼] Step 1 screen
T:0.0
P2
[EDIT/ENTER] P2
ST 1
T:0.0
[1][EDIT/ENTER] P2
ST 1
T:1.0
1: PURGE ON
[EDIT/ENTER] Time flashes (waiting for input) Input 1 minute
T:1.0
RANGE:16 [
][
Function flashes (waiting for input)
P2 ST 1
T:1.0
RANGE:8
ST 2
T:0.0
Time flashes (waiting for input)
P2
ST 2
T:2.0
1: PURGE ON
Input 2 minutes Function flashes (waiting for input)
Call up response [RSPNS/5][EDIT/ENTER] or [ ] four times [EDIT/ENTER]
Value flashes (waiting for input)
] [EDIT/ENTER]
P2
[2][EDIT/ENTER]
Call up sensitivity [RANGE/4][EDIT/ENTER] or [ ] three times [EDIT/ENTER] P2 ST 1
Step 2 screen
ST２ T:0.0
Input 8
P2 ST 2
RESPONSE [
Step 1 input complete
T:2.0
][
STD
] [EDIT/ENTER]
P2 ST 2
T:2.0
RESPONSE FAST
Value flashes (waiting for input) 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 Purge valve open
Key [P-ON/1]
Change keys
Setting range
Purge valve close
[P-OFF/2]
Auto-zero
[A.ZERO/3]
Range
[RANGE/4]
[ ][ ]
[RSPNS/5]
[ ][ ]
1/4,1/2,1,2,4,8,16,32,64, 128,256,512,S FAST, STD, SLOW
Response
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
Time flashes (waiting for input) [1][EDIT/ENTER] P5
[EDIT/ENTER]
ST 3
Time input
T:1.0
1: PURGE OFF
Function flashes (waiting for input)
[RANGE/4][EDIT/ENTER]
PRG2
ST 3
T:1.0
RANGE : 16 [ PRG2
ST 3
][
Sensitivity call up
Value flashes (Change the value here)
][EDIT/ENTER]
T:1.0
RANGE : 32
Figure 6.6 Correcting a Time Program
6.4.2.3 Deletion Single step deletion 1) Display the step to be deleted using the [ ] key. 2) Set the time in the step to 0 ([EDIT/ENTER][0][EDIT/ENTER]). 3) Press the [MONIT] key to return to the monitor screen. Steps will be rearranged in chronological order and the 0 time step will be deleted. Deletion of a step and all subsequent steps 1) Display the first step to be deleted using the [ ] key. 2) Press [SHIFT][CLEAR]. The designated step and all subsequent steps will be deleted.
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”
“MEASURE RANGE” and output scale Dynamic range
RANGE
Inte. output
STD
~ 5x10-4RIU
1/4~512x10-6RIU
WIDE
~ 5x10-3RIU
1/4~512x10-5RIU
5x10-4RIU/1.0V 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 -0.000 [MONIT]
64
[MONIT]
STD
+ ON(OFF)
TEMP: xx.x/yy °C -0.000 + ON(OFF)
[SHIFT] [MARK]
PARAMETER SET 1:MEASURE RANGE [MONIT]
[EDIT/ENTER]
MEASURE RANGE STD(WIDE)
Flashes
[EDIT/ENTER] [
][
MEASURE RANGE
][EDIT/ENTER]
(WIDE) is the setting after the change.
STD
(STD or WIDE can be selected) Input screen
MEASURE RANGE setting 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 5x10-3 RIU(“MEAUSRE RANGE”: WIDE), the measurable range is ~ 7x10-4 RIU/1.0V (“MEASURE RANGE”: STD ), and ~ 9.9x10-3RIU/1.0V (“MEASURE RANGE”: WIDE). When exceeding the linear range, connect to the recorder output terminal instead of the integrator output terminal. For example, when the RANGE setting is 128, the measurable range is 12.8x10-4 RIU/100mV (“MEASURE RANGE”: STD), and 12.8x10-3 RIU/100mV (“MEASURE RANGE”: WIDE).
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)
Tubing
1
2
3
4
5
6
Sample Out (waste)
Flow rate: ~ 10mL/min.
OD=1.6mm ID= 0.8mm Sample IN (from column)
Figure 8.1 “ ~ 10mL/min” tubing arrangement
43
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) Sample
Reference IN
Tubing OD=1.6mm ID=0.8mm (Same tubing as used to connect ports No.1-2 or
1
2
3
4
5
6
ports No.4-5. for the “~10mL/min” tubing
Reference Out
arrangement.
Sample IN Tubing OD=1.6mm, ID=0.8mm
Sample Out
1
2
3
4
5
6
Tubing OD=1.6mm
Mobile phase
ID=0.8mm Replace solvent in the reference cell by sucking the mobile phase solvent.
Sample IN
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
ROM CHECK ERROR
RAM operation
RAM CHECK ERROR
C-MOS RAM backup
BACK UP ERROR
Temperature sensor
TEMP SENSOR NG
Zero glass driver
Z. GLASS ERROR
Lamp operation time
WARNING LAMP MAINTE. TIME
9.1.2 Self-diagnostics during operation The instrument checks the following items during operation (Table 9.2). When a problem is detected, an error message is displayed on the screen. Refer to section 9.1.3 for countermeasures.
Table 9.2 Self-diagnostics Items and Corresponding Error Messages Check item
Error message
Fluid leak in cell
TROUBLE LEAK IN CELL
Heater overheating
TROUBLE OVER TEMP
Light intensity deterioration
WARNING INTENSITY POOR
Optical zero adjustment
WARNING OUT OF RANGE
Lamp operation time
WARNING LAMP MAINTE TIME
9.1.3 Error message meanings and countermeasures 9.1.3.1 TROUBLE LEAK IN CELL This message appears when fluid is leaking from the flow cell. Immediately stop the pump, turn the power off, and contact your local JASCO distributor.
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
Port No.3 RI-2031 Pump
Wash fluid Port No.6 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 saccharose solution is 160x10-6RIU greater than the refractive index of the water. Note:
Use the same water for the mobile phase and to dissovle the saccharose. If different water is used, the correct refractive index may not be obtained.
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 1.0x10-7RIU/min or less. Note:
On the LCD display, 0.1x10-6RIU/min or less corresponds to a 0.1 RIU value per minute. When a recorder is used, this corresponds to approximately 1% or less during a 10 minutes period with a range of 64 (x10-6).
(4) Pump solvent at a flow rate of 1.0mL/min and confirm that the drift is within the limit described in (3) above. In addition, confirm that the fluctuation in the RIU value on the LCD display is less than 0.2.
9.4.4 Calibration method
(1) Following the procedure described in Figure 5.7(refer to section 5.7), display the K1 and K2 Calibration menu screen and press the [EDIT/ENTER] key. The K1 and K2 calibration screen is displayed.
54
Normal operation mode monitor screen NORM -0.000
64
STD
[MONIT]
TEMP: xx.x/yy °C -0.000 + ON(OFF)
+ ON(OFF)
[MONIT]
[SHIFT] [MARK] [ ][ ][EDIT/ENTER]
PARAMETER SET 4:CALIB K [MONIT]
[EDIT/ENTER]
K CALIB: xxxx.x PRESS RUN KEY
K CALIB:
Calibration screen
Once the baseline has stabilized, press the [AUTOZERO] key. Inject standard solution
0.X
PRESS RUN KEY
Read the RIU value shown on the screen for the plateau of the peak. Press the [PRGM RUN] key to execute the built in calibration program. The RIU value will be 160±1 RIU.
K CALIB: 160.0 PRESS RUN KEY
Figure 9.6 Calibration screen (2) In the calibration screen, once the baseline has stabilized, press the [AUTO ZERO] key to set the RIU value to zero. Set the chart speed to 10mm/min. (3) Inject standard solution. (4) If the RIU value on the plateau of peak is within 160.0 ± 1.6 x 10-6 RIU, the instrument is already correctly calibrated. If the value is not within 160.0 ± 1.6 x 10-6 RIU, press the [PRGM RUN] key to execute build-in calibration program. The RIU value will automatically be set to 160.0 ± 1.6 x 10-6 RIU.
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 within 160.0 ± 1.6 x 10-6 RIU without pressing the [PRGM RUN] key. Note:
Adjustment of the integrator or recorder output is not necessary. The outputs will automatically be adjusted when the K1 and K2 values are calibrated.
Note:
Calibration of constants K1 and K2 is performed by executing build-in calibration program at the same time.
9.4.5 Calibration constant confirmation and setting
After performing calibration (outlined in previous section), the calibration constants, K1 and K2 obtained should be recorded. These values should be written on a piece of paper and attached inside the instrument. This way, the recorded values can then be re-input in the following situations, and troublesome calibration operations can be avoided. When confirming and setting the calibration constant, follow the procedure shown in Figure 9.8 and re-input the calibration constants. (1) When the K1 or K2 values have been changed by mistake
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 -0.000
[MONIT]
64
+ ON(OFF)
-0.000
[
][
+ ON(OFF)
]
[
][
[EDIT/ENTER]
]
PARAMETER SET 3:SET K2 VALUE [MONIT]
K1 VALUE: x.xxx RIU: xxxx.x [MONIT]
TEMP: xx.x/yy°C
[SHIFT] [MARK]
PARAMETER SET 2:SET K1 VALUE [MONIT]
[MONIT]
STD
[EDIT/ENTER]
K2 VALUE: x.xxx RIU: xxxx.x
[EDIT/ENTER]
[MONIT]
[EDIT/ENTER]
K2 VALUE: x.xxx RIU: xxxx.x
K1 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 -0.0
64
[MONIT]
TEMP: xx.x/yy °C -0.0
+ ON(OFF)
+ ON(OFF)
[SHIFT] [MARK]
[MONIT] (Use the [
STD
]/[
] keys to display the menu below)
Lamp maintenance time menu PARAMETER MODE 2:L. MAINTE. TIME [MONIT]
[EDIT/ENTER]
LAMP MAINTE TIMER 10000 H
[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 [MONIT]
NORM 64 STD -0.0 + OFF
[MONIT]
TEMP: xx.x/yy°C -0.0 + OFF
[SHIFT] [MARK]
(Use the [
]/[
] keys to display the menu below)
Serial number menu PARAMETER MODE 6:SERIAL NO. [MONIT]
[EDIT/ENTER]
SERIAL NO 1234567890
[EDIT/ENTER] [xxxxxxxxxx] [EDIT/ENTER]
SERIAL NO 1234567890 Flashes
Figure 9.10 Serial Number Input Method
9.7 Power Fuse replacement Warning:
When changing the power fuses, be sure to disconnect the power cable from the power cable inlet (refer to Figure 2.5).
Warning:
For continued protection against risk of fire, replace only with fuses of the specified type and current ratings.
Note:
When one fuse is blown, replace both fuses.
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
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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.
nS
i
i
θ r nR
Figure 10.1 Horizontal Cross-section of Flow Cell and Light Path The area to the left of the center partition is called the sample cell and the area to the right is called the reference cell. ns : Refractive index of sample nR : Refractive index of reference (solvent) θ: Angle of deflection (exaggerated in the figure: actual value is 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
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