Professional Documents
Culture Documents
HLC-8320GPC
Operators Manual
Revision G
Tosoh Corporation
Bioscience Division
Safety Precautions
Introduction
Operators who use this instrument should read the safety precautions first and handle
the instrument correctly.
WARNING and CAUTION are defined below. For safe use, be sure to observe the
warnings and cautions.
At installation
II
Safety Precautions
On use
Do not touch the power plug or main power switch with wet hands.
Touching the power plug or main power switch with wet hands may result in electric
shock.
III
Safety Precautions
While the instrument is running, keep the covers and doors closed.
You may be caught or pulled in by movable or heat generating sections or by
high-voltage electric circuits inside the instrument and suffer a burn or electric
shock.
IV
Safety Precautions
V
Safety Precautions
VI
Safety Precautions
When using a column switching valve, note the flow rate at switching.
When an optional column switching valve is equipped to use columns of different
sizes, valve switching without changing the flow rate may damage the column. Set
an appropriate flow rate for the column size in advance.
When the rack of the auto sampler is out, keep your hands away from
the cover.
If your hand is near the cover of the auto sampler when rack is out, your hand may
be struck by the cover, or be caught or pulled into the instrument, causing serious
injury.
VII
Safety Precautions
Be sure there is no leak in tubing. Leak of organic
CAUTION
solvent may cause corrosion, fire, electric shock or
poisoning.
CAUTION Keep away from an open flame. Organic solvent may
catch fire and explode.
VIII
Safety Precautions
Notes
No part of this document may be quoted or reproduced in any form without prior
permission from TOSOH.
This document is subject to revision without notice.
Tosoh Corporation
Bioscience Division
IX
About This Document
This operators manual describes how to use and operate this instrument
correctly. This document is based on the following rules:
Section title
For sections that exceed
one page, the font size of
the section title is small
on the second and later
pages.
Illustration
Illustrations are provided
to help the reader to
understand the text.
Symbols
The symbols described
below indicate situations
in which care is required
during operation.
Page
Page numbers are by chapter.
X
Contents
Chapter 2 Operation
2.1 Operation Outline 2-2
2.2 Opening or Closing the Oven Doors 2-5
2.3 Turning the Power ON or OFF 2-6
2.4 Setting the Operation-display 2-7
2.5 Viewing the Monitor Screen 2-9
2.6 Stop Valve 2-11
2.7 Purge Function 2-12
2.8 Pump Drain 2-13
2.9 Changing the Degasser Vacuum Pressure 2-14
2.10 Changing the Degasser Volume 2-15
2.11 Controlling the Pump Unit 2-17
2.12 Controlling the Temperature Control 2-20
2.13 Controlling the Auto Sampler 2-22
2.14 Controlling the Detector (RI) 2-28
2.15 Setting the Warm-up Conditions 2-30
2.16 Executing Warmup 2-32
2.17 Setting the Shutdown Conditions 2-33
2.18 Executing Shutdown 2-35
2.19 Setting the Sample Queue 2-36
2.20 Preparing Samples 2-38
2.21 Loading Sample Cups 2-39
2.22 Starting and Stopping Analysis 2-40
2.23 Weekly Timer 2-41
2.24 Setting the Clock 2-42
XI
Contents
Chapter 3 Analysis
3.1 Analysis Notes 3-2
3.2 Analysis Flow Chart 3-4
Chapter 6 Troubleshooting
6.1 Checking Errors 6-2
6.2 Error Messages 6-3
6.3 Before Seeking Repairs 6-13
XII
Contents
Chapter 7 Installation
7.1 Installation Environment 7-2
7.2 Installation Place 7-3
7.3 Installation Space 7-5
Chapter 8 Connection
8.1 Connecting the Power Cable 8-2
8.2 Removing the Auto Sampler Fixture 8-3
8.3 Connecting a PC 8-4
8.4 Tubing 8-5
8.5 Attaching the Washing Tubing to the Rear of the Pump 8-12
8.6 Installing the Cases 8-13
8.7 Connecting the I/O Terminals 8-14
Chapter 9 Specifications
9.1 Specifications 9-2
Chapter 10 Options
10.1 UV Absorption Detector (UV-8320) 10-2
10.2 External Detector 10-10
10.3 Column Switching Valve 10-11
XIII
Contents
Chapter 12 Theory
12.1 Instrument Configuration 12-2
12.2 Pump Unit 12-3
12.3 Auto Sampler 12-4
12.4 Detector (RI) 12-5
12.5 Detector (UV) 12-7
Chapter 13 References
13.1 Solvent Miscibility 13-2
13.2 Solvent Characteristics 13-3
13.3 Quick Reference for Boiling Point 13-8
13.4 Waste Liquid Treatment 13-9
13.5 Countermeasures against Static Electricity 13-10
XIV
Chapter 1
Auto sampler
Degas/Suction unit
Pump oven
1-2
1.2 Sides
Left side
If the outlets of the drain tubing and wash drain tubing are fixed higher than
those on the left side of the instrument, the solvent will not be drained
smoothly but will overflow from the drain block or washing port. For smooth
drainage, cut the tubing to appropriate lengths before putting them into a
waste container so that the ends of the tubing will be low.
1-3
1.2 Sides
Right side
I/O terminals
See 1.5, I/O Terminals and Communication Port.
1-4
1.3 Rear
Back-pressure release
tubing
1-5
1.4 Operation-display
POWER key
(Power ON/OFF)
Function keys
(For functions displayed in
the bottom line)
Control keys 1 Control keys 2
(For executing and indicating (For executing and indicating
warmup, analyze, and sample rack ejection, sampler
shutdown) wash, auto balance, sample
pump flow, reference pump flow,
and purge)
1-6
1.4 Operation-display
Control keys 1
Start/stop warmup
WARMUP
The key lamp remains lit during execution. To stop warmup, press
and hold the key for one second.
Start/stop analysis
ANALYZE
The key lamp remains lit during execution and flashes to indicate
reservation. If this key is pressed during warmup, analysis will start
after warmup. To cancel or stop analysis after finishing the sample
that has already been injected, press and hold the key for one
second. For forced termination, press and hold the key again in this
state for one second.
Start/stop shutdown
SHUTDOWN
The key lamp remains lit during execution and flashes to indicate
reservation. If this key is pressed during analysis, shutdown will start
after analysis is complete. To cancel or abort shutdown, press and
hold the key for one second.
Control keys 2
Eject/insert rack
This key is used to eject or insert a sample rack. If rack is inserted ,
the LED lamp lights.
Start/stop purge
PURGE
The key lamp remains lit during execution. To specify the capacity
by numeric keypad, press and hold the key for one second. If the
key is held for one second during execution, purge stops.
1-7
1.5 I/O Terminals and Communication Port
RI terminal
This terminal outputs analog signals from the RI detector.
Output value: Integrator range/1V
UV terminal
This terminal outputs analog signals from the UV
detector (option).
Output value: Integrator range/1V
INJ terminal
This signal outputs a relay contact signal at sample
injection by the auto sampler.
ERROR terminal
Connect a contact output terminal of external
instrument to this terminal. This instrument execute
error processing after signal input.
See P6-4 Error No.0022
PC terminal
Connect a USB cable to this terminal to control this
instrument and acquire data by the PC control program
EcoSEC WorkStation.
1-8
1.6 Inside
Pump oven
Inline filters
Check valves
UV detector (optional) housing
Sample pump
Reference pump
1-9
1.6 Inside
Degas/Suction unit
1-10
1.6 Inside
Column oven
R.COL
S.COL
1-11
MEMO
1-12
Chapter 2
Operation
2.1 Operation Outline
This manual explains how to control the instrument from the operation-display. If a
PC is connected to this instrument by a USB cable, disconnect the USB cable.
The figure below shows the transition diagram when this instrument is controlled from
the operation-display.
( Page )
Process Screen
( Seq. )
( Page )
Sample queue
( Ana. ) screen
( Page )
( Page )
( ) is displayed in the bottom line. Press the function key below the displayed
message to go to the corresponding screen.
2-2
2.1 Operation Outline
( Page )
Unit selection
(Param.) screen
( Unit ) ( Page )
( Unit ) ( Page )
Tube volume
(TubeV.) measurement screen
( Unit )
Oven parameter
screen
( Unit )
Column switching
valve screen
( Unit )
RI detector
parameter screen
( Unit ) ( Page )
UV detector UV detector
parameter screen 1 ( Page ) parameter screen 2
Lamp intensity
(Power ) check screen
*1 In case that a column switching valve (option) is equipped, the display changes to a screen
according to the selected column.
Column 1: Pump parameter screen 1
Column 2: Pump parameter screen 2
*2 In case that column switching valve (option) is not equipped, the display changes to Pump
parameter screen 3.
2-3
2.1 Operation Outline
( Page )
Option screen
(Option)
( Page )
2-4
2.2 Opening or Closing the Oven Doors
[Opening]
1 Lift the OPEN buttons (B) at the bottom right of the pump oven door and at the
bottom left of the column oven door.
2 The doors open.
[Closing]
1 Press CLOSE (A) securely at the center of each door to close the doors.
If the two doors are not locked securely, temperature control fails.
2-5
2.3 Turning the Power ON or OFF
This instrument has the main power switch on the right side. Turn the switch to ON ( | )
to start using the instrument.
If the instrument will not be used for a long time, set the main power switch to OFF ({).
This will completely stop the power supply to the instrument.
------ ON
------ OFF
When the main power switch is OFF ({), the instrument cannot be operated
from a PC.
Once the main power switch has been turned OFF ({), do not turn it ON ( | )
again within five seconds.
Power ON/OFF
The Power ON/OFF can also be controlled from the PC control program EcoSEC
WorkStation.
2-6
2.4 Setting the Operation-display
By introducing actual screens, this section explains how to set the operation-display.
For operating procedures and detailed functions of this instrument, see the
corresponding sections.
[Monitor screen 1]
( ) is displayed in the bottom line. Press the function key below the displayed
message to go to the corresponding function screen.
Ex. Press below (Param.) to go to the unit selection screen.
Select Unit : 3
1: Pump 2: AS 3: Oven
4: Column Change 5: RI 6: UV 7: VIS
( Back )
The cursor is _ on the screen. Type in a figure from the numeric keys and press the
ENT key for confirmation.
ENT
Ex. Press and to go to the oven parameter screen.
CLR
Press to return to the initial value or the value before editing.
Press BACK
to erase the numeral at the cursor, and the cursor returns to the
previous digit.
2-7
2.4 Setting the Operation-display
If there are plural input items to one screen , the cursor moves to the next input item
each time the ENT key is pressed.
< > is displayed in the bottom line. Press the function key below the displayed
message to execute the corresponding function screen.
Ex. Press below <CO Ctr> to start temperature control of the column oven.
2-8
2.5 Viewing the Monitor
The monitor screen consists of two pages. Press (Page) to change pages.
[Monitor screen 1]
UV output (option) Sample pump set flow rate
RI output UV wavelength (option) Sample pump pressure
2-9
2.5 Viewing the Monitor
[Monitor screen 2]
2-10
2.6 Stop Valve
The solvent supply to a instrument can be blocked by switching the stop valve.
For maintenance and inspection such as the replacement of a check valve, switch the stop
valve to the stop position.
2-11
2.7 Purge Function
The purge function primes a solvent by a dedicated syringe pump. This function is used to
replace a solvent or to degas from a pump.
If PURGE
is pressed and held for one second while purge is not running, the purge screen
is displayed. To change the purge volume, enter an arbitrary value at "Volume" on this
screen and press ENT
. The input range is from 1 to 150 mL.
[Purge screen]
Purge : STOP
Volume : 10
( Back )
2-12
2.8 Pump Drain
For switching to the drain channel, only the sample side or reference side pump can be
switched.
2-13
2.9 Changing the Degasser Vacuum Pressure
This instrument has a degasser. Degasser control automatically starts at flow or purge.
If fluorine or any other solvent of great permeability through the PTFE film is used in the
degasser, change the vacuum pressure to Low.
2-14
2.10 Changing the Degasser Volume
The degasser volume on the solvent side is determined by two systems of 20 mL. Change
the degasser volume according to the flow rate. The reference capacity is 20 mL when the
total flow rate of the sample and reference pumps is less than 1.0 mL/min and is 40 mL
when it is 1.0 mL/min or more.
Before changing the degasser volume, turn the main power OFF. Since
some sections are driven by motors, fingers or objects may be caught or
pulled into the instrument, causing serious injury.
2-15
2.10 Changing the Degasser Volume
4 When setting the degasser volume to 40 mL, make the following connections:
(WASH)
Solvent suction
tubing
To stop valve
5 When setting the degasser volume to 20 mL, make the following connections:
Solvent suction
To stop valve
tubing
2-16
2.11 Controlling the Pump Unit
Flow start/stop
S.FLOW
1 To start the flow of the sample pump, press . The LED lamp lights ON
and S.Pump on Monitor screen 1 changes from STOP to Set flow rate.
R.FLOW
2 To start the flow of the reference pump, press . The LED lamp lights ON
and R.Pump on Monitor screen 1 changes from STOP to Set flow rate.
3 To stop the flow of the sample pump, press and hold S.FLOW for one second.
The LED lamp lights OFF and S.Pump on Monitor screen 1 changes from Flow
rate to STOP.
4 To stop the flow of the reference pump, press and hold R.FLOW for one second.
The LED lamp lights OFF and R.Pump on Monitor screen 1 changes from Flow
rate to STOP.
Control the pump unit and change their settings as explained below.
First, display Pump parameter screen 1 by the following procedure:
The pump parameter screen consists of four screens. The screen changes each
time (Page) is pressed.
2-17
2.11 Controlling the Pump Unit
Sam. Flow When changing the flow rate of the sample pump, enter an arbitrary
numeric value at Sam.Flow and press ENT
.
The input range is from 0.010 to 2.000 mL.
Ref. Flow When changing the flow rate of the reference pump, set Ref.Flow in the
format of 1/X of the sample pump. Enter a numeric value from 1 to 4
and press ENT . The actual flow rate is displayed in ( ).
P.Limit When changing the upper or lower pressure limit, enter an arbitrary
numeric value at P.Limit and press ENT . The upper pressure limit is
the sample side, and the lower pressure limit is the reference side. The
input range of lower limit is from 0.0 to 24.9 MPa. The input range of
upper limit is 0.1 to 25.0 MPa. And the lower limit must be smaller than
the upper limit.
2-18
2.11 Controlling the Pump Unit
For the column switching method, see 10.3, Column Switching Valve.
1 Measure the actual flow rate when flowing at the set flow rate.
2 Check the current setting at Control Flowrate.
3 Enter the measured flow rate at Measure Flowrate and press ENT .
4 Press (Calc.) to display a correction coefficient at Flowrate Coeff.
5 A correction efficient may also be entered directly at Flowrate Coeff. Enter an
arbitrary value and press ENT . The input range is from 0.800 to 1.200.
For the flow rate measuring method, see 4.1, Checking the Pump Unit
Performance.
2-19
2.12 Controlling the Temperature Control
Temperature control of the pump oven and column oven and change their settings as
described below.
First, display the oven parameter screen by the following procedure:
CO Temp. When changing the set temperature of the column oven, enter an
arbitrary numeric value at CO Temp. and press ENT . The input range
is from 30 to 60C.
Set the value of the room temperature +10C or higher.
The actually measured value is displayed in ( ).
PO Temp. When changing the set temperature of the pump oven, enter an arbitrary
numeric value at PO Temp. and press ENT . The input range is from
30 to 50C.
Set the value of the room temperature +10C or higher.
The actually measured value is displayed in ( ).
2-20
2.12 Controlling the Temperature Control
Gas When changing the gas sensor threshold, enter an arbitrary value at
Gas and press ENT . The input range is from 1 to 1,000. The upper
item for the column oven and the lower item is for the pump oven.
The actually measured value is displayed in ( ).
Be careful that tubing or other objects do not enter the hole of a partition
in the pump oven or column oven. This is very dangerous because of a
fan rotating inside.
The gas sensor sensitivity differs with the solvent. To ensure accurate
gas detection, check the gas sensor sensitivity with the solvent to be
used in advance and then check the threshold. A greater displayed value
(gas sensor measured value) indicates a higher gas concentration.
For the gas sensor, record a value measured in an ordinary state and a value
measured when a wiper containing approximately 0.5 mL of a solvent is
inserted into the oven. Then set the intermediate value as the threshold.
Set each oven to room temperature +10C or higher.
For stable flow, the pump oven controls the temperature of the pump unit.
Since a solvent having a low boiling point may make flow unstable, turn the
temperature control OFF for such solvents.
Do not set the temperature in a range where the solvent concentration
changes greatly. For THF, 40C is recommended.
2-21
2.13 Controlling the Auto Sampler
Washing
1 Press WASH
to start washing. The LED lamp lights ON.
2 If WASH is pressed and held for one second, the manual washing volume can
be changed. Enter a volume using the numeric keys and press ENT . The input
range is from 1 to 20,000 L. Manual Wash V. on Sampler parameter screen 1 is
updated with the input value.
Control the auto sampler and change its settings as described below.
First, display Sampler parameter screen 1 by the following procedure:
The sample parameter screen consists of three pages. The page changes
each time (Page) is pressed.
Change the injection parameters on the sample queue screen.
2-22
2.13 Controlling the Auto Sampler
At shipping, the syringe speed is set to 6 for Sam. Speed and 14 for Wash
Speed. When using THF as the solvent, the settings need not be changed.
When using a solvent of higher viscosity or measuring a sample of great
viscosity, reduce the speed. The relationship between input values and actual
speeds is as follows:
Input value 1 2 3 4 5 6 7 8
Speed (L/s) 5 10 15 20 30 40 50 65
Input value 9 10 11 12 13 14 15
Speed (L/s) 80 100 125 180 250 350 500
For ordinary needle washing, the washing volume from approximately 500 to
1,000 L is sufficient. For solvent replacement, however, set the manual
washing volume to 20,000 L.
2-23
2.13 Controlling the Auto Sampler
2-24
2.13 Controlling the Auto Sampler
1 Press (AirS.) on Sampler parameter screen 1 to display the air sensor sensitivity
adjustment screen.
2 Press <Meas.> to measure the transmissive light strength of air and solvent.
The intermediate value between the maximum and minimum is calculated.
3 Press (Set) to confirm the value of Step 2 as the threshold.
When the difference between the maximum and minimum is below 100, the
air sensor may not function well at analysis. Wash the sampler thoroughly and
execute measurement again.
Instead of automatic adjustment, manual input is possible. Enter arbitrary
numeric value after Set, and press ENT .
2-25
2.13 Controlling the Auto Sampler
For the sample loop replacement method, see 5.8, Replacing the Sample
Loop.
From the viewpoint of the linearity of injection volume and the carry over, we
recommend the use of a sample loop less than 500 uL.
2-26
2.13 Controlling the Auto Sampler
The row of 1 to 20 indicates the status of Cup No. 1 to 20 sequentially from the left. By
pressing (Up) or (Down), the status of Cup No. 1 to 100 can be checked. c indicates a
cup with no sample shortage or no injection and indicates a cup having a sample
shortage.
In the above figure, Cup No. 13 and 25 were lacking a sample.
2-27
2.14 Controlling the Detector (RI)
Auto balance
Press BALANCE . The detector output signal moves to the set balance. UV auto
balance is executed only when an optional UV detector is equipped.
2-28
2.14 Controlling the Detector (RI)
RESP. Set the time constant of RI detector output Enter 1 (0.5 s), 2 (1.0 s),
or 3 (3.0 s) and press ENT .
<Temp> Turn optical block temperature control ON or OFF. When the
temperature control is turned ON, the RI.Temp LED lamp lights.
<NGlass> To do the optical auto balancing.
The response is changed according to the half-value width of the peak. The
table gives the reference response for the half-value width of each peak.
2-29
2.15 Setting the Warmup Conditions
[Warmup screen 1]
Column: 1(Column1) UVLamp: 1(ON)
Purge: 1(EXEC) 10
WarmupFlow%: 50 (0.300 0.300)
( Page ) ( Back )
If liquid is supplied at the analysis flow rate when the column temperature is still
low at the start of warmup, the column may be deteriorated by a great pressure
difference in the column.
Set the flow rate at warmup (WarmupFlow%) lower than the analysis flow rate
(approximately 50% is recommended).
2-30
2.15 Setting the Warm-up Conditions
[Warm-up screen 2]
CO: 1 (ON ) PO: 1 (ON ) RI: 1 (ON )
WarmupTime : 15
SteadyTime : 0
( Page ) ( Back )
2-31
2.16 Executing Warmup
To start warmup, press WARMUP on the operator panel. The LED lamp remains lit during
execution. If WARMUP is pressed and held for one second when the LED lamp is lit,
warmup is finished immediately.
2-32
2.17 Setting the Shutdown Conditions
[Shutdown screen 1]
AfterSeq.Time : 10
ShutdownFlow% : 50(0.300 0.300)
CO: 0 (OFF) PO: 0 (OFF) RI: 0 (OFF)
( Page ) ( Back )
If temperature control is stopped at the analysis flow rate when analysis using the
column oven is finished, the column may be deteriorated by a great pressure
difference in the column. If the instrument is stopped immediately after analysis, a
temperature drop may cause the solvent to shrinkage and flow back into the
column. After analysis, stop the temperature control of the column oven and keep
flow at a lower flow rate than the analysis flow rate (approximately 50% is
recommended) until the column cools down.
2-33
2.17 Setting the Shutdown Conditions
[Shutdown screen 2]
ShutdownTime: 30
Power: 0 (OFF)
( Page ) ( Back )
ShutdownTime Set the time from the start of shutdown until flow stops. Enter a
ENT
numeric value from 0 to 999 [min] and press .
Power Select whether to turn the power OFF at the end of shutdown. Enter
0 (OFF) or 1 (ON) and press ENT
.
2-34
2.18 Executing Shutdown
SHUTDOWN
To start shutdown, press on the operator panel. The LED lamp remains lit
during execution. If the key is pressed and held for one second when the LED lamp is
lit, shutdown is suspended.
If the SHUTDOWN
is pressed during analysis, shutdown is reserved and the
LED lamp starts flashing. When AfterSeq.Time has passed after analysis,
shutdown is executed .
Even during warmup, shutdown can be reserved as during analysis.
By using the weekly timer function, shutdown can be executed at a specified
time on a specified day of the week. However, analysis has priority and
shutdown is not executed during analysis.
If analysis is aborted by an error, error processing has priority and shutdown
is not executed.
2-35
2.19 Setting the Sample Queue
[Process screen]
RunTime: 15.0
Balance : 0 (OFF)
RunTime Enter the sample injection interval. Enter a numeric value from 3 to
999.9 [min] and press ENT .
Balance Select whether to execute auto balance automatically.
Enter 0(OFF)/1(INJ)/2(DAC) and press ENT .
OFF : No auto balance
INJ : Auto balance just before sample injection
DAC : Auto balance just before data acquisition (when the
EcoSEC-WorkStation is not used, the timing is the
same as that for INJ).
2 When specifying an injection cup, press (Ana.) to display the sample queue
screen.
2-36
2.19 Setting the Sample Queue
Repeat Enter the number of repetitive injections from a single sample cup.
The input range is from 1 to 9 [times]. Enter a value after checking the
injection volume, the presuction volume, and the injection volume.
Inj.Vol. Enter the injection volume. The input range is from 1 to 1,500 [L].
Wave. Enter the wavelength of the UV detector. The input range is from 195 to
350 [nm].
If the setting is changed here, the wavelength is changed just before the
injection.
This is displayed only when an optional UV detector is used.
( Up ) Move the cursor one line above.
Scroll the screen if there are more lines above.
( Down ) Move the cursor to one line below.
Scroll the screen if there are more lines below.
( Ins. ) Add to the current line. The setting one line above is copied and
inserted.
( Del. ) Delete the current line.
Set the auto balance timing where the baseline is stable. For analysis
(overlap injection) shown in the figure below, auto balance at the injection
timing is actually executed at the negative peak position. If auto balance is
executed there, the baseline position will deviate greatly.
2-37
2.20 Preparing Samples
Five types of sample cups are prepared for this instrument. (See 11.2, Consumables.)
If evaporation causes a sample concentration problem, seal a sample cup with an
aluminum sheet or a septum packing, as shown below. The aluminum sheet or septum
packing is used once only because it will have a hole after sample injection.
It is recommended to place as much of the sample as possible because bubbles may be
loaded if the sample is not sufficient.
If evaporation causes a sample concentration problem in repetitive injection from a single
sample cup, use a septum-type cup.
The instrument may not load a sample of high viscosity. Change the syringe
speed or dilute the sample.
At suction, the needle does not reach the bottom of a cup. Place a sample to
10 mm or higher from the bottom of a cup.
2-38
2.21 Loading Sample Cups
Eject
Insert
After sample injection, the sample rack may be ejected until the next sampling
operation starts. If the next sampling time comes when the sample rack is
ejected (not inserted correctly), the instrument will pause.
When inserting the sample rack, push the rack in until it clicks. However,
excess force may damage the instrument.
When the sample rack is inserted correctly, the LED lamp lights.
2-39
2.22 Starting and Stopping Analysis
2-40
2.23 Weekly Timer
W.up Select whether to reserve warmup with the timer by the day of the week.
Enter 0 (OFF) or 1 (ON) and press ENT .
Enter the warmup start time.
S.down Select whether to reserve shutdown with the timer by the day of the
week. Enter 0 (OFF) or 1 (ON) and press ENT .
Enter the shutdown start time.
(Page) Change the screen. Page 1 is for Monday to Wednesday, Page 2 for
Thursday to Saturday, and Page 3 for Sunday.
When using a weekly timer, keep the main power ON. Also, check the
warmup and shutdown conditions well before use.
If warmup or analysis is in progress at a specified warmup time, priority is
given to the current processing.
If analysis is in progress at a specified shutdown time, priority is given to the
current processing. Shutdown is executed after the analysis.
2-41
2.24 Setting the Clock
( Set ) ( Back )
Now Displays the current date, day of the week, and time.
Set Enter a day of the week and time to set.
Enter 0 (SUN) to 6 (SAT) for a day of the week and time by the
24-hour system.
(Set) Confirm the setting.
2-42
2.25 Checking the Instrument Program Version
The version of the program installed in the instrument can be checked as follows:
[Option screen]
PRG Main : 99.99
Sub1 : 99.99
Sub2 : 99.99
(E.Log ) ( Back )
2-43
2.26 Checking Error Logs
This instrument stores the most recent nine error logs. They can be checked as follows:
The error history is displayed in order of date, time, and error number.
By changing (Page), you can check up to nine error logs.
For the error numbers and their contents, see Chapter 6, Troubleshooting.
If the EcoSEC WorkStation is connected, all error logs can be stored and
checked.
2-44
Chapter 3
Analysis
3.1 Analysis Notes
Solvent replacement
When replacing a solvent, check that the solvents are miscible with each other.
If the solvents are not miscible, substitute the current solvent with a miscible solvent
and then with the target solvent.
Tubing washing
After a salt-containing solvent or a buffer solution is used, wash the tubing with
deionized water or ion-exchanged water. Without washing, salt may precipitate and
disable further use.
If a chlorine-based organic solvent such as chloroform is used, wash the tubing with
ethanol or isopropanol. If the tubing is not washed, rusting may make accurate
analysis impossible.
3-2
3.1 Analysis Notes
If the plunger seal or plunger is dry, be sure to purge it before flow start. (See 2.7,
Purge Function.)
When not using the instrument for one week or longer, remove the column, wash the
tubing, and replace the solvent with ethanol or isopropyl before storage.
Static electricity
When an inflammable organic solvent is used as a solvent, static electricity may cause
a fire or explosion.
To prevent an accident, the accumulation of static electricity should be suppressed.
(See 13.5, Countermeasures against Static Electricity.)
3-3
3.2 Analysis Flow Chart
This section explains the operating procedure of this instrument from power-on until
power-off.
Set sample queue Set the sample cup number, injection volume, injection
interval, and injection count for the sample queue.
(See 2.19, Setting the Sample Queue.)
3-4
Chapter 4
Performance Check
4.1 Checking Pump Unit Performance
[Conditions]
Flow rate : 0.5 mL/min
Solvent : Deionized water (degassed)
Load : capillary (inner diameter: 0.1 mm, length: 4 m)
[Procedure]
1 Open the column oven door and connect a stainless steel tubing to the
channel on the sample and reference side.
2 Replace the solvent in the system with deionized water and start flow.
3 When the flowing pressure has become stable, collect the discharge liquid
from the sample and reference sides in a measuring cylinder or measuring
flask and measure the time required to collect 5 mL of water. Repeat this
procedure three times.
4 Calculate the actual flow rate as follows:
Actual flow rate = 5 mLMeasuring time (min)
5 Determine the maximum, minimum, and average actual flow rates from the
measurement results and calculate the flow rate precision as follows:
A B
Flow rate precision = 100 (%)
B
A: Maximum or minimum actual flow rate
B: Average actual flow rate
6 Check that the flow rate precision is within 0.4%.
Before replacing the solvent in the system, check the solvent miscibility.
[Procedure]
1 From the average actual flow rate obtained by the above measurement,
calculate the flow rate accuracy as follows:
BC
Flow rate accuracy = 100 (%)
C
B: Average actual flow rate
C: Set flow rate
2 Check that the flow rate accuracy is within 4.0%.
4-2
4.2 Checking Pump and Column Oven Performance
[Procedure]
1 Set the temperatures of the pump oven and column oven at 40C.
(Room temperature: 20 to 25C)
2 Start temperature control of each oven.
3 After 90 min, check the temperature displayed on the monitor.
4 Check that the temperature of each oven is within 400.2C.
[Procedure]
1 With the temperature control ON, place a wiper containing approximately 0.5
mL of THF in the oven and close the door.
2 Check that liquid leakage occurs and temperature control stops.
3 With the oven door open, allow the THF gas to sufficiently dissipate from the
oven.
The gas sensor sensitivity differs with the solvent. Check in advance
that the gas sensor detects the gas to use.
4-3
4.3 Checking Detector (RI) Performance
Sensitivity check
[Conditions]
Sample : 0.67 g of sucrose dissolved in 1 liter of deionized water
Injection volume : 500 L
Flow rate : 0.2 mL/min (both sample and reference)
Range : 256
Response (RI) : 0.5 s
Polarity :
Control temperature : 40C
Column : None
Data acquisition time : 0 - 5 min
Sampling interval : 100 ms
Data analysis condition : Column test
[Procedure]
1 Replace the sample loop of the auto sampler with the 500 L sample loop.
(See 5.8, Replacing the Sample Loop.)
2 Replace the solvent in the detector (RI) cell (both sample and reference) with
deionized water.
3 Under the above conditions, inject the sample into the cell and record the RI
output signal.
4 In the peak editor for the EcoSEC WorkStation data analysis, set the baseline
as shown below and check that the peak height is within 390 20 mV.
Output
Within
390 mV20 20
390 mV mV
4-4
4.3 Checking Detector (RI) Performance
Be sure to check that the peak is flat at the top. The cell must be saturated
with a sample for an accurate sensitivity check.
If the tubing volume between the sample loop and the cell is greater than the
initial volume because an optional column switching valve is used, the cell
may not be saturated with a sample. In this case, remove the valve tubing
and check again.
If the EcoSEC WorkStation is not used, connect an external instrument
(integrator or recorder) through the RI terminal and check that the output
peak height is within 390 20 mV.
4-5
4.4 Checking Auto Sampler Performance
[Conditions]
Washing liquid and solvent : Deionized water (degassed)
Sample : 0.67 g of sucrose dissolved in 1 liter of
deionized water
Injection volume : 10 L
Flow rate : 0.5 mL/min (both sample and reference)
Load : Capillary (inner diameter: 0.1 mm, length: 4 m)
Range : 256
Response (RI) : 0.5 s
Control temperature : 40C
Injection interval : 4 min
Data acquisition time : 0 - 3 min
Sampling interval : 50 ms
Data analysis condition : Column test
[Procedure]
1 Under the above conditions, prepare two sample cups on a sample rack,
inject the sample five times (10 times) per cup, and record the peak height.
2 Measure each peak height and calculate CV.
(xi x )
2
s=
n 1
s
CV = 100(%)
x
s : Standard deviation
xi : Measured value
x : Average value
n : Data count
CV : Coefficient of variation
3 Check that CV is 1.5 or less.
4-6
4.5 Comprehensive Inspection
Drift check
[Conditions]
Solvent : THF
Range : 256
Response (RI) : 1.0 s
Control temperature : 40C
Polarity :
Data acquisition time : 0 - 120 min
Sampling interval : 1,000 ms
[Procedure]
1 Turn on the power to the instrument, start temperature control, and flow THF
to both the sample and reference sides at a flow rate of 1 mL/min.
2 Under the above conditions, record the detector (RI) baseline for two hours.
3 Measure the one-hour baseline fluctuation from one hour after power-on and
check that the fluctuation is within 5107RIUh (within 2 mV).
60min
4-7
4.5 Comprehensive Inspection
Noise check
[Conditions]
Range : 256
Response (RI) : 0.5 s
Control temperature : 40C
Polarity :+
Data acquisition time : 0 - 5 min
Sampling interval : 100 ms
[Procedure]
1 After checking the drift, stop flow.
2 After the baseline becomes stable, record the detector (RI) baseline for five
minutes under the above conditions.
3 Divide the baseline into 30-second segments. Measure the noise width in
each segment and obtain the average value of the 10 segments.
4 Check that the average noise width is within 2.510-8RIU (within 0.1 mV).
10
Y
R=1
X1 X2 X3 X4 X5 X6 X7 X8 X9 X10
Y2 Y6 Y7 Y8
Y3 Y5 Y9 Y10
Y1
Y4
5 min
4-8
Chapter 5
If pulsation or pressure fluctuation increases during flow, wash the check valves.
[Necessary items]
Wrench 810 mm (Part No. ----------) 1 Accessory
Wrench 1214 mm (Part No. ----------) 1 Accessory
Internal-type union (Part No. 0007337) 2
Finger tight fitting (Part No. 0016566) 2
Deionized water or ethanol Appropriate amount
Solvent bottle 1
Beaker 1
Capillary (Part No. 0005319) 1
[Procedure]
1 Stop flow.
2 Remove the column (if attached) and attach the capillary.
3 Replace the solvent in the system with deionized water or ethanol.
4 Close the stop valve. (See 2.6, Stop Valve.)
5 With a wrench, disconnect the tubing from the check valves.
6 With a wrench, remove the check valves from the pump head.
Check valve
(Outlet)
Check valve
(Inlet)
5-2
5.1 Washing the Check Valves
For solvent replacement in the system, check the solvent miscibility in advance.
5-3
5.2 Replacing the Check Valves
If the pulsation or pressure fluctuation is great even after the check valves of the pump unit
are washed, replace the check valves.
[Necessary items]
Check valve assembly (Outlet) (Part No. 0022012) Necessary quantity
Check valve assembly (Inlet) (Part No. 0022011) Necessary quantity
Wrench 810 mm (Part No. ----------) 1 Accessory
Wrench 1214 mm (Part No. ----------) 1 Accessory
Internal-type union (Part No. 0007337) 2
Fingertight fitting (Part No. 0016566) 2
Deionized water or ethanol Appropriate amount
Solvent bottle 1
Capillary (Part No. 0005319) 1
[Procedure]
1 Stop flow.
2 Remove the column (if attached) and attach the capillary.
3 Replace the solvent in the system with deionized water or ethanol.
4 Close the stop valve. (See 2.6, Stop Valve.)
5 With a wrench, disconnect tubing from the check valves.
6 With a wrench, remove the check valves from the pump head.
7 With a wrench, attach new check valves to the pump head.
8 With a wrench, connect the tubing.
9 Execute purge and replace deionized water or ethanol in the system with an
used solvent.
Check that there is no leakage from tubing joints. (See 2.7, "Purge Function.")
For solvent replacement in the system, check the solvent miscibility in advance.
5-4
5.3 Replacing the Plunger Seal
If solvent still leaks from the washing block behind the pump head, replace the plunger
seal.
[Necessary items]
Plunger seal (Part No. 0019190) Necessary quantity
Wrench 810 mm (Part No. ----------) 1 Accessory
Hex wrench 4 mm (Part No. ----------) 1 Accessory
Plunger seal replacement tool (Part No. 0018524, 0018525) 1 set Accessory
Internal-type union (Part No. 0007337) 2
Finger tight fitting (Part No. 0016566) 2
Deionized water or ethanol Appropriate amount
Solvent bottle 1
Capillary (Part No. 0005319) 1
[Procedure]
1 Stop flow.
2 Remove the column (if attached) and attach the capillary.
3 Replace the solvent in the system with deionized water or ethanol.
4 Close the stop valve. (See 2.6, Stop Valve.)
5 With a wrench, remove the tubing from the pump head.
6 With the 4 mm hex wrench, loosen the cap screw at the center of the pump
head holder and remove the right and left mounting screws.
7 Pull the pump head straight out to remove.
Check the plunger for no damage. Replace the plunger if damaged.
5-5
5.3 Replacing the Plunger Seal
Plunger seal
Pump head
Collar
Mounting screw
Washing block
Cap screw
(Only loosen)
8 Remove the plunger seal from the pump head using the guide bar of plunger
seal replacement tool, as shown below.
Guide bar
Plunger seal
Pump head
5-6
5.3 Replacing the Plunger Seal
9 Set a new plunger seal on the guide bar of plunger seal replacement tool as
shown below.
10 Insert the guide bar into the pump head. (Note the plunger seal insertion
direction.)
11 Hold the guide bar upright and push the plunger seal into the pump head with
the seal holder.
12 While holding the seal holder, extract the guide bar straight.
13 Remove the seal holder.
Seal holder
Plunger seal
Guide bar
Pump head
14 Push the pump head straight in. With the 4 mm hex wrench, attach the right
and left mounting screws to the pump head holder and tighten the cap screw at
the center.
15 With a wrench, connect the tubing.
16 Execute purge and replace the deionized water or ethanol in the system with
an used solvent.
Check that there is no leakage from tubing joints. (See 2.7, "Purge Function.")
For solvent replacement in the system, check the solvent miscibility in advance.
5-7
5.4 Replacing the Plunger
If solvent still leaks from the washing block even after the plunger seal at the pump unit is
replaced, replace the plunger.
[Necessary items]
Plunger (Part No. 0019762) Necessary quantity
Wrench 810 mm (Part No. ----------) 1 Accessory
Wrench 1214 mm (Part No. ----------) 1 Accessory
Hex wrench 3 mm (Part No. ----------) 1 Accessory
Hex wrench 4 mm (Part No. ----------) 1 Accessory
Internal-type union (Part No. 0007337) 2
Finger tight fitting (Part No. 0016566) 2
Deionized water or ethanol Appropriate amount
Solvent bottle 1
Capillary (Part No. 0005319) 1
[Procedure]
1 Stop flow.
2 Remove the column (if attached) and attach the capillary.
3 Replace the solvent in the system with deionized water or ethanol.
4 Close the stop valve. (See 2.6, Stop Valve.)
5 With a wrench, remove tubing from the pump head.
6 With the 4 mm hex wrench, loosen the cap screw at the center of the pump
head holder and remove the right and left mounting screws.
7 Pull the pump head straight out to remove.
8 With the 3 mm hex wrench, remove the washing block mounting screws.
9 Pull the washing block straight out to remove.
10 With a wrench, remove the old plunger and attach a new plunger.
11 When the plunger is removed, check the diaphragm for scratches or cracks. If
scratch or crack is found, replace the diaphragm (Part No. 019056).
5-8
5.4 Replacing the Plunger Seal
Diaphragm
Plunger
Washing block
Plunger seal
Collar
Mounting screw
(large)
Cap screw
(Only loosen)
12 Push the washing block straight in and attach the washing block mounting
screws with the 3 mm hex wrench.
13 Push the pump head straight in. With the 4 mm hex wrench, attach the right
and left mounting screws to the pump head holder and tighten the cap screw at
the center.
14 With a wrench, connect the tubing.
15 Execute purge and replace the deionized water or ethanol in the system with
an used solvent.
Check that there is no leakage from tubing joints. (See 2.7, "Purge Function.")
5-9
5.5 Washing or Replacing the Inline Filter
If the pressure becomes higher at flow, the inline filter may be stopped up. Wash or replace
the inline filter.
[Necessary items]
Inline filter element (Part No. 0017714) Necessary quantity
Wrench 1417 mm (Part No. ----------) 2 Accessory
Wrench 810 mm (Part No. ----------) 1 Accessory
Internal-type union (Part No. 0007337) 2
Finger tight fitting (Part No. 0016566) 2
Deionized water or ethanol Appropriate amount
Solvent bottle 1
Beaker 1
Capillary (Part No. 0005319) 1
[Procedure]
1 Stop flow.
2 Remove the column (if attached) and attach the capillary.
3 Replace the solvent in the system with deionized water or ethanol and check
the pressure.
4 Close the stop valve. (See 2.6, Stop Valve.)
5 With a wrench, remove the tubing from the top of the inline filter.
6 With a wrench, remove the inline filter.
7 Soak the removed inline filter in a beaker containing deionized water or ethanol
and wash them by ultrasonic cleaning. If the filter element has scratches,
cracks, or stains, replace the filter element.
8 With a wrench, attach the inline filter.
9 With a wrench, connect the tubing to the top of the inline filter.
10 Execute purge and replace deionized water or ethanol in the system with an
used solvent.
Check that there is no leakage from tubing joints. (See 2.7, "Purge Function.")
5-10
5.5 Washing or Replacing the Filter
<Inline filter>
Inline filter
Press in
Assembly
Tighten with
wrench
For solvent replacement in the system, check the solvent miscibility in advance.
5-11
5.6 Replacing the 6way Valve Rotor Seal and Stator
If the reproducibility of injection volume is not well or if liquid leaks from the needle, check
the rotor seal and stator of the 6way valve (sample injection valve) for damage. Replace
the rotor seal and stator if damaged. When changing the rotor seal and stator of the
sampler 4way valve at the same time, see 5.7, Replacing the 4way Valve Rotor Seal and
Stator.
[Necessary items]
Rotor seal(6way) (Part No. 0016415) Necessary quantity
6way Valve for Stator (Part No. 0016451) Necessary quantity
Wrench 810 mm (Part No. ----------) 1 Accessory
Wrench 1/45/16 (Part No. ----------) 1 Accessory
Hex wrench 3 mm (Part No. ----------) 1 Accessory
Beaker 1
Deionized water or ethanol Appropriate amount
Capillary (Part No. 0005319) 1
[Procedure]
1 Stop flow.
2 Remove the column (if attached) and attach the capillary.
3 Replace the solvent in the system with deionized water or ethanol.
4 Close the stop valve. (See 2.6, Stop Valve.)
5 With a wrench, remove the tubing (other than sample loop) from Ports 2, 3, 5,
and 6 of the 6way valve.
6 With a hex wrench, remove the three bolts securing the stator.
For solvent replacement in the system, check the solvent miscibility in advance.
Sliding
5-12
5.6 Replacing the 6way Valve Rotor Seal and Stator
7 Remove the stator from the valve body and check the sliding surface of the
stator. Only if damage is found, replace the stator.
8 If no damage is found, soak the removed stator in a beaker containing
deionized water or ethanol and then wash it by ultrasonic cleaning.
When removing the sample loop for stator replacement, attach the stator to the
valve body.
If the rotor seal is difficult to remove, drive two M3 screws into the holes of the
rotor seal.
Rotor seal
M3 screw
5-13
5.6 Replacing the 6way Valve Rotor Seal and Stator
3: Column
4: Sample loop
2: Pump
5: 4way valve
1: Sample loop
6: Needle
5-14
5.7 Replacing the 4way Valve Rotor Seal and Stator
If the reproducibility of injection volume is not well or if liquid leaks from the needle, check
the rotor seal and stator of the sampler 4way valve (pressure relief valve) for damage.
Replace the rotor seal and stator if damaged. When changing the rotor seal and stator of
the 6way valve (sample injection valve) at the same time, see 5.6, Replacing the 6way
Valve Rotor Seal and Stator.
Before changing the degasser volume, turn the main power OFF. Since
some sections are driven by motors, fingers or objects may be caught or
pulled into the instrument, causing serious injury.
[Necessary items]
Rotor seal(4way) (Part No. 0016414) Necessary quantity
4way Valve Stator (Part No. 0016450) Necessary quantity
Wrench 810 mm (Part No. ----------) 1 Accessory
Wrench 1/45/16 (Part No. ----------) 1 Accessory
Hex wrench 3 mm (Part No. ----------) 1 Accessory
Beaker 1
Deionized water or ethanol Appropriate amount
Capillary (Part No. 0005319) 1
[Procedure]
1 Turn the main power OFF.
2 Stop flow.
3 Remove the column (if attached) and attach the capillary.
4 Replace the solvent in the system with deionized water or ethanol.
5 Close the stop valve. (See 2.6, Stop Valve.)
6 Open the sampler syringe unit on the left side of the main unit.
7 With a wrench, disconnect the tubing from the sampler 4way valve ports.
8 With a hex wrench, remove the three bolts securing the stator.
5-15
5.7 Replacing the 4way Valve Rotor Seal and Stator
For solvent replacement in the system, check the solvent miscibility in advance.
When removing the sample loop for stator replacement, attach the stator to the
valve body.
If the rotor seal is difficult to remove, drive two M3 screws into the holes of the
rotor seal.
Rotor seal
M3 screw
5-16
5.7 Replacing the 4way Valve Rotor Seal and Stator
3: 6way valve
5: Degasser chamber
5-17
5.8 Replacing the Sample Loop
This instrument is equipped with a 100-L sample loop. If necessary, replace the sample
loop with a loop of appropriate volume.
[Necessary items]
Wrench 1/45/16 (Part No. ----------) 1 Accessory
Deionized water or ethanol Appropriate amount
Capillary (Part No. 0005319) 1
[Procedure]
1 Stop flow.
2 Remove the column (if attached) and attach the capillary.
3 Replace the solvent in the system with deionized water or ethanol.
4 Loosen the male nuts at Ports 1 and 4 of the 6way valve in the column oven
and remove the sample loop.
5 Attach the sample loop for replacement. Tighten the male nuts while pressing
the sample loop deep inside the port.
For solvent replacement in the system, check the solvent miscibility in advance.
After replacing the sample loop, change the setting of the loop volume.
For details, see 2.13, Controlling the Auto Sampler.
From the viewpoint of the linearity of injection volume and the carry over, we
recommend the use of a sample loop less than 500 uL.
5-18
5.9 Washing the Detector (RI) Cell
[Necessary items]
Internal-type union (Part No. 0007337) 4
Finger tight fitting (Part No. 0016566) 4
Capillary (Part No. 0005319) 2
Waste container 1
When aqueous liquid, saline solution,
or buffer solution is used: Deionized water Appropriate amount
When organic solvent is used: Ethanol or isopropanol Appropriate amount
[Procedure]
1 Stop flow.
2 Remove the column (if attached).
3 Connect a capillary to the sample column IN tubing (S.COL) and the detector
(RI) sample IN tubing (RI.S).
4 Connect a capillary to the reference column IN tubing (R.COL) and the
detector (RI) reference IN tubing (RI.R).
5 Supply the washing liquid from the solvent suction tubing (SOLVENT) at a flow
rate of 1 mL/min for 10 minutes or longer.
6 Replace the washing liquid in the system with an used solvent and flow for 10
minutes or longer.
For solvent replacement in the system, check the solvent miscibility in advance.
5-19
5.10 Washing the Inside of the System
[Necessary items]
Internal-type union (Part No. 0007337) 4
Fingertight fitting (Part No. 0016566) 4
Capillary (Part No. 0005319) 2
Deionized water Appropriate amount
Nitric acid (Regulation of 1 to 3) Appropriate amount
Solvent bottle 2
Waste container 1
[Procedure]
1 Stop flow.
2 Remove the column (if attached).
3 Connect a capillary to the sample column IN tubing (S.COL) and the detector
(RI) sample IN tubing (RI.S).
4 Connect a capillary to the reference column IN tubing (R.COL) and the
detector (RI) reference IN tubing (RI.R).
5 Replace the solvent in the system with deionized water.
6 Replace the solvent with nitric acid.
7 Flow at a flow rate of 1 mL/min. The reference flow time is 2 hours in the case
of Regulation 1 and 40 min in the case of Regulation 3.
8 Replace the nitric acid in the system with deionized water and flow for 10
minutes or longer.
9 Replace the washing liquid in the system with an used solvent and flow for 10
minutes or longer.
When using nitric acid, be sure to wear protective items (gloves and
goggles). If nitric acid gets on your skin, wash it away thoroughly by
flowing water.
For solvent replacement in the system, check the solvent miscibility in advance.
5-20
Chapter 6
Troubleshooting
6.1 Checking Errors
If an error occurs in this instrument, a buzzer will sound and the operation-display will
display an error screen.
Check the error number, error message, and error information (Info 1 to 3). For
troubleshooting, refer to the following pages.
When consulting TOSOHs service engineer, report the error number, error message, and
error information 1 to 3.
After checking the error contents, press <Buzzer> to stop the buzzer and (Back) to return
to Monitor screen 1. The buzzer will also stop if (Back) is pressed.
[Error screen]
The user can check up to nine logs on this instrument. For checking error
logs, see 2.26, Checking Error Logs.
When the PC control program EcoSEC WorkStation is connected, the PC
displays an error message. In addition, all error logs can be stored for later
checking.
6-2
6.2 Error Messages
Error contents
0001 USB Communication Error Turn the main power OFF and 0
USB command ID error occurred. contact TOSOH service engineer.
0002 USB Communication Error Turn the main power OFF and 0
USB data error occurred. contact TOSOH service engineer.
0003 USB Communication Error Turn the main power OFF and 0
USB timing error occurred. contact TOSOH service engineer.
0013 Master System Load Error Turn the main power OFF and 0
The installed program could not be contact TOSOH service engineer.
read. (Master)
6-3
6.2 Error Messages
0014 Slave1 System Load Error Turn the main power OFF and 0
Could not load the slave 1 program. contact TOSOH service engineer.
0015 Slave2 System Load Error Turn the main power OFF and 0
Could not load slave 2 program. contact TOSOH service engineer.
0016 Slave1 Trans Error Turn the main power OFF and 0
Error occurred in the transmission to contact TOSOH service engineer.
slave1 ROM from master ROM.
0017 Slave1 Receive Error Turn the main power OFF and 0
Error occurred in the reception to contact TOSOH service engineer.
slave1 ROM from master ROM.
0018 Slave2 Trans Error Turn the main power OFF and 0
Error occurred in the transmission to contact TOSOH service engineer.
slave2 ROM from master ROM.
0019 Slave2 Receive Error Turn the main power OFF and 0
Error occurred in the reception to contact TOSOH service engineer.
slave2 ROM from master ROM.
0020 Backup Error (Param) Turn the main power OFF and then 0
The backup loss error of the parameter ON again. Then confirm the settings.
occurred. If the error recurs, turn the main
power OFF and contact TOSOH
service engineer.
0021 Backup Error (Adjust) Turn the main power OFF and 0
The backup loss error of the instrument contact TOSOH service engineer.
adjustment value occurred.
0022 External Error Check the connected external 3
The error signal was inputted from instrument (integrator or recorder).
external instrument.
0023 Slave1 Communication Error Turn the main power OFF and 0
The internal communication error contact TOSOH service engineer.
occurred. There is no response from a
Slave1.
6-4
6.2 Error Messages
0024 Slave2 Communication Error Turn the main power OFF and 0
The internal communication error contact TOSOH service engineer.
occurred. There is no response from a
slave2.
0025 Slave1 Trans Buffer Full Turn the main power OFF and 0
The internal communication error contact TOSOH service engineer.
occurred. The sending buffer of slave1
ROM overflowed.
0026 Slave2 Trans Buffer Full Turn the main power OFF and 0
The internal communication error contact TOSOH service engineer.
occurred. The sending buffer of slave2
ROM overflowed.
0027 Power Fan Stop Turn the main power OFF and 0
The fan of a power supply has stopped. contact TOSOH service engineer.
0200 Pump (Sam) Pressure High Check the tubing of the sample 3
Pressure upper limit error occurred in channel for no clogging.
the sample pump.
0201 Pump (Sam) Pressure Low Check the tubing of the sample 3
Pressure lower limit error occurred in channel for no leakage.
the sample pump.
0202 Pump (Sam) Motor Error Turn the main power OFF and then 3
Sample pump drive error occurred. It ON again.
may be a cam gap. If the error recurs, turn the main
power OFF and contact TOSOH
service engineer.
0203 Pump (Ref) Pressure High Check the tubing of the reference 3
Pressure upper limit error occurred in channel for no clogging.
the reference pump.
0204 Pump (Ref) Pressure Low Check the tubing of the reference 3
Pressure lower limit error occurred in channel for no leakage.
the reference pump.
6-5
6.2 Error Messages
0205 Pump (Ref) Motor Error Turn the main power OFF and then 3
Reference pump drive error occurred. It ON again.
may be a cam gap. If the error recurs, turn the main
power OFF and contact TOSOH
service engineer.
0207 Purge Syringe Home Error Turn the main power OFF and then 3
Purge syringe drive error occurred. ON again.
If the error recurs, turn the main
power OFF and contact TOSOH
service engineer.
0208 Drain Valve Position Error Turn the main power OFF and then 3
Drain valve drive error occurred. ON again.
If the error recurs, turn the main
power OFF and contact TOSOH
service engineer.
0209 Stop Valve Position Error Turn the main power OFF and then 3
Stop valve drive error occurred. ON again.
If the error recurs, turn the main
power OFF and contact TOSOH
service engineer.
6-6
6.2 Error Messages
0300 Injection Valve Position Error Turn the main power OFF and then 3
6way (injection) valve drive error ON again.
occurred. If the error recurs, turn the main
power OFF and contact TOSOH
service engineer.
0301 Syringe Valve Position Error Turn the main power OFF and then 3
4way (syringe) valve drive error ON again.
occurred. If the error recurs, turn the main
power OFF and contact TOSOH
service engineer.
0302 Sample Syringe Home Error Turn the main power OFF and then 3
Syringe drive error occurred. ON again.
If the error recurs, turn the main
power OFF and contact TOSOH
service engineer.
0303 Sample Rack Home Error Turn the main power OFF and then 3
Sample rack drive error occurred. ON again.
If the error recurs, turn the main
power OFF and contact TOSOH
service engineer.
0304 Sample Arm Home Error Turn the main power OFF and then 3
Sampling arm drive error occurred. ON again.
If the error recurs, turn the main
power OFF and contact TOSOH
service engineer.
0305 Sample Needle Home Error Turn the main power OFF and then 3
Sampling needle drive error occurred. ON again.
If the error recurs, turn the main
power OFF and contact TOSOH
service engineer.
0306 Sample Short( This Cup Skip In Future) Add a sample as required and 0
Sample shortage. execute measurement again.
6-7
6.2 Error Messages
0309 Sample Rack Eject Time Out This warning is issued if the rack is 2
The sample rack is left out. Please left out for 10 minutes or longer.
insert the rack. Insert the rack.
0400 Column Change Valve Position Error Turn the main power OFF and then 3
Valve drive error occurred at column ON again.
switching. If the error recurs, turn the main
power OFF and contact TOSOH
service engineer.
0402 Oven (CO) Temp. High Check that the set temperature is 3
Temperature control upper limit error the room temperature +10C or
occurred in the column oven. higher.
Temperature control stopped because Turn the main power OFF and
the temperature exceeded the set contact TOSOH service engineer.
temperature +7C.
0403 Oven (CO) Temp. Low Check that the oven door is closed 3
Temperature control lower limit error securely.
occurred in the column oven. If the error recurs, turn the main
power OFF and contact TOSOH
service engineer.
0404 Oven (CO) Temp. Heat Error This error occurs when temperature 3
Temperature rise error occurred in the has not reach the control range after
column oven. 1 h from control start. Check that the
oven door is closed securely.
If the error recurs, turn the main
power OFF and contact TOSOH
service engineer.
6-8
6.2 Error Messages
0405 Oven (CO) Temp. High Limit Turn the main power OFF and 3
The column oven temperature contact TOSOH service engineer.
exceeded 70C and the upper
temperature limit error occurred.
0406 Oven (CO) Gas Leak If there is any leak, wipe off the 3
Gas leak error occurred in the column solvent and keep the door open for
oven. a time to allow ventilation.
If this error occurs without any leak,
adjust the gas sensor.
0408 Oven (CO) Door Open Close the door to start temperature 2
The column oven door is open. Close control.
the door to start temperature control.
0409 Oven (PO) Temp. High Check that the set temperature is 3
Temperature control upper limit error the room temperature +10C or
occurred in the pump oven. higher.
Temperature control stopped because Turn the main power OFF and
the temperature exceeded the set contact TOSOH service engineer.
temperature +7C.
0410 Oven (PO) Temp. Low Check that the oven door is closed 3
Temperature control lower limit error securely.
occurred in the pump oven. If the error recurs, turn the main
power OFF and contact TOSOH
service engineer.
0411 Oven (PO) Temp. Heat Error This error occurs when temperature 3
Temperature rise error occurred in the has not reach the control range after
pump oven. 1 h from control start. Check that the
oven door is closed securely.
If the error recurs, turn the main
power OFF and contact TOSOH
service engineer.
6-9
6.2 Error Messages
0414 Oven (PO) Door Open Close the door to start temperature 2
The pump oven door is open. Close the control.
door to start temperature control.
0415 Oven (CO) Temp. Sensor Error Contact TOSOH service engineer. 3
Column oven master sensor
disconnection error.
0416 Oven (CO) Temp. Sensor Error Contact TOSOH service engineer. 3
Column oven slave sensor
disconnection error.
0417 Oven (PO) Temp. Sensor Error Contact TOSOH service engineer. 3
Pump oven sensor disconnection error.
6-10
6.2 Error Messages
0502 RI Over Flow Error Flow solvent untill Over Flow Error is 0
RI Over Flow Error removed.
0603 UV Grating Home Error Turn the main power OFF and 3
Grating drive error occurred in UV contact TOSOH service engineer.
detector.
6-11
6.2 Error Messages
0803 Set Time Error (Acquire > Runtime) Check the setting and ensure 0
The acquisition time cannot be set (acquisition end time - acquisition
beyond the runtime. Please check the start time) < runtime.
parameters.
6-12
6.3 Before Seeking Repairs
If this instrument does not operate normally, perform the following checks:
turn ON. Is the main power switch ON Turn the main power switch ON
()? ().
Execute purge.
The flow pressure is Is air trapped?
(See 2.7, "Purge Function.")
not stable.
Wash or replace the check
valves. (See 5.1, Washing the
Are the check valves clean?
Check Valves and 5.2,
Replacing the Check Valves.)
6-13
6.3 Before Seeking Repairs
6-14
6.3 Before Seeking Repairs
6-15
6.3 Before Seeking Repairs
6-16
Chapter 7
Installation
7.1 Installation Environment
Install this instrument on a level desk free of harmful gas, dust, or vibration where it is not
exposed to direct sunlight or wind and use it under the following environmental conditions:
Temperature 15 to 35C
Humidity 40 to 80% (No condensation)
Altitude 2,000 m max
Dust General office environment
Power supply AC 100 to 240 V, 50/60 Hz, 500 VA
When moving this instrument, need two or more people and hold the
bottom right and left of the instrument.
7-2
7.2 Installation Place
7-3
7.2 Installation Place
7-4
7.3 Installation Space
The outside dimensions of this instrument are 680W 580H 500D mm. To place the
accessory solvent tray (width: 240 mm), an installation space of approximately 1,000 650
mm is necessary.
When using the EcoSEC WorkStation PC control program, installation space for a personal
computer (PC) is also necessary.
680 mm
240 mm
650 mm
PC
Tray HLC-8320GPC
1,000 mm
Since this instrument has a cooling fan blow port on the back, be
sure to allow for a space of approximately 100 mm behind the
instrument.
This instrument weighs 95 kg. Considering the weight, install the
instrument in a stable place.
7-5
MEMO
7-6
Chapter 8
Connection
8.1 Connecting the Power Cable
1 Check that the main power switch on the right side of this instrument is OFF ().
2 Plug in one end of the power cable to the power inlet on the right side of this
instrument.
3 Plug in the other end of the power cable to a power socket with a ground.
When connecting the power cable, check that the main power switch on
the right side of this instrument is OFF ().
8-2
8.2 Removing the Auto Sampler Fixture
At the shipping of this instrument, the drive section of the auto sampler is secured.
When using this instrument, remove the fixture by the following procedure:
Before removing the fixture, turn the main power OFF and remove the
power cable.
8-3
8.3 Connecting a PC
When using the PC control program EcoSEC WorkStation to control this instrument and to
acquire data, connect a PC to the PC terminal on the right side of this instrument with a USB
cable.
Before connecting, add ferrite cores to both side of the USB cable.
With the PC connected, turn ON the main power to this instrument in order to activate the
acquisition control application and to find the instrument. For details, refer to the operators
manual of the EcoSEC WorkStation.
Right side
8-4
8.4 Tubing
Tubing types
The tubing described below protrudes from the left side of this instrument:
8-5
8.4 Tubing
If the drain tubing, or the wash drain tubing of the auto sampler, or the waste container
is positioned or installed at an elevated position relative to the instrument, liquid
leakage may occur in the instrument.
Install the waste container lower than the tubing outlet of this instrument.
Ensure that the drain tubing remains lower than the tubing outlet.
8-6
8.4 Tubing
The solvent bottle cap is connected by the two methods explained below. Select the
connection that is suitable for use.
A. Ordinary connection
B. Connection to use a salt-containing solvent or a buffer solution
A. Ordinary connection
1 Attach fittings and adapters to the wash suction tubing (WASH) and solvent suction
tubing (SOLVENT) and connect them through the inner cap. Adjust the tubing so
that it protrudes approximately 30 cm beyond the inner cap and fasten the fittings to
secure the tubing. Pass the wash suction tubing (WASH) through the
small-diameter port and pass the solvent suction tubing (SOLVENT) through the
large-diameter port.
2 Attach the suction filter to the end of the solvent suction tubing (SOLVENT) having
an outer diameter of 1/8.
3 Pass the PTFE tubing of the prevention kit of moisture adsorption through the cap
hole and connect it to the large-diameter port.
4 Attach the solvent bottle end plug to the small-diameter port.
Wash suction
Solvent bottle end plug
O.D. 1/16" Cap tubing (WASH)
Solvent suction
tubing (SOLVENT)
8-7
8.4 Tubing
Inner cap
HLC-8320GPC
WASH
SOLVENT
8-8
8.4 Tubing
Column connection
1 Connect the S.COL tubing from Port 3 of the 6way valve to the inlet of the analytical
column. Connect the outlet of the analytical column to the RI.S tubing.
2 Connect the R.COL tubing of the pump unit to the inlet of the reference column.
Connect the outlet of the reference column to the RI.R tubing.
R.COL
S.COL
8-9
8.4 Tubing
Each column has a tag indicating the grade and the flow direction. When
connecting a column, check the flow direction.
When connecting more than one column, arrange them as shown below.
If the tubing is too short, extend the inlet tubing (valve to column tubing). If the
tubing is connected by a joint such as a union, the performance may
deteriorate. It is recommended that new tubing be created.
Flow direction
Valve
Valve
Column tag
RI.S RI.S
8-10
8.4 Tubing
H/HXL/HHR Series
h = 4 mm
Screw type
8-11
8.5 Attaching Washing Tubing to the Rear of the Pump
If the solvent contains salt, the salt may precipitate from the plunger seal of the pump unit
and reduce the life of the plunger seal. To prevent this problem, fill the diaphragm section
with deionized water for periodic washing.
[Necessary items]
Deionized water Approximately 200 mL
Silicon tubing 422 m (Part No. 008851) 1
Disposable syringe 20 mL (Part No. 008571) 1
8-12
8.6 Installing the Cases
Install the accessory solvent tray on the left side of the instrument to hold the solvent bottle
and waste container.
The ovens of this instrument have gas sensors. If liquid leakage occurs in an oven, the
temperature control and the flow stop. Since the instrument is designed to drain leaking fluid
from the bottom, install the accessory instrument tray at the bottom of the instrument.
Solvent tray
Instrument tray
Insert from the front of the
instrument until it hits the stopper.
The pump oven and the column oven have drain ports to deal with liquid leakage
if any.
The auto sampler and the degas/suction unit are designed to drain leaking liquid
from the bottom if it should leak.
8-13
8.7 Connecting the I/O Terminals
This instrument has I/O terminals and a communication port on the right side. When using
external instrument (integrator or recorder), connect it to the I/O terminals.
The EcoSEC WorkStation can be used only by USB cable connection.
1 Remove the two screws from the terminal window on the right side of the
instrument, place the cover as shown below, and secure it with a screw.
ferrite core
(parts No.022001)
Right side
8-14
8.7 Connecting the I/O Terminals
RI terminal
This terminal outputs a 25610-6 or 6410-6 RIU/1V signal from the detector (RI). By
connecting the input terminal of external instrument (integrator or recorder) here
through an analog signal cable (Part No. 0017546), data can be processed.
UV terminal
This terminal outputs a 0.5, 1.0, 2.0, or 4.0 ABU/1V signal from the detector (UV). By
connecting the input terminal of external instrument (integrator or recorder) here
through an analog signal cable (Part No. 0017546), an output signal from the detector
(UV) can be recorded.
This is valid only when the optional UV-8320 is equipped.
INJ terminal
This terminal outputs a relay contact signal at sample injection by the auto sampler. By
connecting the start signal terminal of an integrator or other external instrument here,
the relay contact signal can be used as a start signal for the instrument.
ERROR terminal
By connecting the contact output terminal of external instrument here, error processing
of this instrument can be executed at signal input. See P6-4 Error No.0022
Cable insertion
tool
8-15
MEMO
8-16
Chapter 9
Specifications
9.1 Specifications
Degasser system
Pump oven
9-2
9.1 Specifications
Pump Unit
Auto sampler
9-3
9.1 Specifications
Column oven
Detector (RI)
9-4
9.1 Specifications
Optional valve
9-5
9.1 Specifications
I/O terminals
PC communication : USB
RI signal : Analog output (-1 to 1 V)
UV signal : Analog output (-1 to 1 V)
Injection signal output : Contact output
Error signal input : Contact or TTL-level input
Stainless steel 316, PTFE, PEEK, zirconia, silica glass, PFA, and carbon-contained
PTFE, Vespel*
*When a column switching valve (option) is used.
Installation environment
Temperature : 15 to 35C
Humidity : 40 to 80% (No condensation)
Altitude : 2,000 m max
Dust : General office environment
9-6
9.1 Specifications
tc VH
VL
VH : 2.0 V min
VL : 0.8 V max
tc : 100 ms min
tc High
Low
NORMAL OPEN
Allowable current :1A
tc : 1.0 to 1.2 s
9-7
MEMO
9-8
Chapter 10
About Options
10.1 UV Absorbance Detector (UV-8320)
Tubing
Connect the IN tubing (UV.IN) of the UV-8320 to the sample column outlet and the
OUT tubing (UV.OUT) of the UV-8320 to the detector (RI) sample IN tubing (RI.S).
UV.OUT
RI.R
10-2
10.1 UV Absorbance Detector (UV-8320)
Control
Press BALANC to move the detector output signal to the set balance value. Auto
balance is executed simultaneously on the RI detector.
Changing settings
Change the settings of the UV-8320 as explained below.
First, display UV detector parameter screen 1 by the following procedure:
10-3
10.1 UV Absorbance Detector (UV-8320)
WAVE Set the wavelength, enter the wavelength by the numeric keys and
press ENT . The input range is from 195 to 350 [nm].
Pola. Set the polarity of UV detector output, change the polarity to output a
negative peak to the positive side. Enter 0 (-) or 1 (+) and press ENT .
Resp. Set the time constant of UV detector output, enter 1 (0.5 s), 2 (1.0 s), or
3 (3.0 s) and press ENT .
<Lamp> Turns the lamp on or off
10-4
10.1 UV Absorbance Detector (UV-8320)
Performance check
Drift check
[Conditions]
Solvent :THF
Wavelength :254 nm
Range : 1 ABU/FS
Response : 1.0 s
Control temperature : 40C
Polarity :+
Data acquisition time : 0-120 min
Sampling interval : 1,000 ms
[Procedure]
1 Turn on the power of the HLC-8320GPC, start temperature control, and flow
THF by the sample pump at a flow rate of 0.35 mL/min.
2 Under the above conditions, record the detector (UV) baseline for two hours.
3 Measure the one-hour baseline fluctuation from one hour after power-on and
check that the drift is within 5104 ABU/h (within 0.5 mV).
60min
10-5
10.1 UV Absorbance Detector (UV-8320)
Noise check
[Conditions]
Range : 1 ABU/FS
Response : 1.0 s
Control temperature : 40C
Polarity :+
Data acquisition time : 0-5 min
Sampling interval : 100 ms
[Procedure]
1 After checking the drift, stop flow.
2 After the baseline becomes stable, record the detector (UV) baseline for five
minutes under the above conditions.
3 Divide the baseline into 30-second segments. Measure the noise width in
each segment and obtain the average value of the 10 segments.
4 Check that the average noise width is within 510-5 ABU (within 0.05 mV).
10
Y
R=1
X1 X2 X3 X4 X5 X6 X7 X8 X9 X10
Y2 Y6 Y7 Y8
Y3 Y5 Y9 Y10
Y1
Y4
5 min
10-6
10.1 UV Absorbance Detector (UV-8320)
Sensitivity check
[Conditions]
Solvent : Deionized water
Sample :0.05% p-toluenesulfonic acid sodium/water
Injection volume : 500 L
Flow rate : 0.5 mL/min (both sample and reference)
Wavelength : 262 nm
Range : 1ABU/FS
Response : 0.5 s
Polarity :+
Control temperature : 40C
Column : None
Data acquisition time : 0-3 min
Sampling interval : 100 ms
Data analysis condition : Column test
[Procedure]
1 Replace the sample loop of the auto sampler with the 500 L sample loop.
(See 5.8, Replacing the Sample Loop.)
2 Replace the solvent with deionized water and wait until the baseline becomes
stable.
3 Under the above conditions, inject the sample into the cell and record the
output signal.
4 In the peak editor for the EcoSEC WorkStation data analysis, set the baseline
as shown below and obtain the peak height (X).
Output
Peak height (X)
10-7
10.1 Ultraviolet Absorbance Detector (UV-8320)
5 Check that the difference between the peak height (X) and the theoretical
output conversion value (Y) = 332 mV is 5 %.
XY
100 < 5
Y
Be sure to check that the peak is flat at the top. The cell must be saturated
with a sample for an accurate sensitivity check.
If the tubing volume between the sample loop and the cell is greater than
the initial volume because an optional column switching valve is used, the
cell may not be saturated with a sample. In this case, remove the valve
tubing and check again.
If the EcoSEC WorkStation is not used, connect an external instrument
through the UV terminal and check that the output peak height satisfies
the above standard.
10-8
10.1 Ultraviolet Absorbance Detector (UV-8320)
Cell washing
[Necessary items]
Finger tight fitting (Part No. :016566) 2
Internal-type union (Part No. :007337) 2
Capillary 1
Waste container 1
When aqueous liquid, saline solution,
or buffer solution is used: Deionized water Appropriate amount
When organic solvent is used: Ethanol or isopropanol Appropriate amount
[Procedure]
1 Stop flow.
2 Remove the column (if attached).
3 Connect a capillary to the sample column IN tubing (S.COL) and the detector
(UV) sample IN tubing (UV.IN).
4 Supply the washing liquid from the solvent suction tubing (SOLVENT) at a
flow rate of 1 mL/min for 10 minutes or longer.
5 Replace the washing liquid in the system with an used solvent and flow for 10
minutes or longer.
10-9
10.2 External Detector
When using a detector other than the UV-8320 as an external detector or when using
a fraction collector, contact TOSOH service engineer.
10-10
10.3 Column Switching Valve
<Flow channel>
Auto sampler
injection valve
5
6
1
4
2 Column switching
3
valve
3 2
1
Analytical column 1 4 Analytical column 2
8
5
6 7
When using a column of a different size, note the flow rate at valve switching.
Since a different flow rate can be set for each column switching valve
position, set the flow rate for Analytical column 2 according to the column.
(See 2.11, Controlling the Pump Unit.)
10-11
10.3 Column Switching Valve
<Example of tubing>
COL1 COL2
250 mm 350 mm
INJ RI.S
135 mm 600 mm
450 mm 600 mm
REF RI.R
10-12
10.3 Column Switching Valve
10-13
MEMO
10-14
Chapter 11
Parts List
11.1 Accessories
If the accessories are purchased as maintenance parts, their quantities may be different.
Inspection certificate 1
Warranty card 1
Power cable 1
Wrench 8 mm 10 mm 1
Wrench 12 mm 14 mm 1
Wrench 14 mm 17 mm 1
11-2
11.1 Accessories
Hex wrench 4 mm 1
11-3
11.1 Accessories
11-4
11.1 Accessories
11-5
11.2 Consumables
019762 Plunger(zirconia) 1
11-6
11.2 Consumables
11-7
11.2 Consumables
11-8
11.2 Consumables
11-9
11.3 Options
11-10
11.3 Options
11-11
11.3 Options
11-12
11.3 Options
PTFE tubing
Part No. Name Standard Quantity
006587 PTFE tubing 2.0 mm.0 mm2 m 1
Special tube
Part No. Name Standard Quantity
016481 Silicon tubing 2.5 mm1.5 mm2 m 1
Material: SUS316
008299 Long nut 1/16 5
Material: SUS316
016180 Ferrule 1/16 10
Material: SUS316
006163 Internal-type union 1/16, 0.35 5
Material: SUS316
11-13
11.3 Options
Material: SUS316
022005 Internal-type union 1/16, 1 5
short
Material: SUS316
007539 T-joint 1/16, 0.4 1
Material: SUS316
007055 T-joint 1/16, 1 1
Material: SUS316
008878 Male nut 1/8 5
Material: SUS316
006176 Ferrule 1/8 10
Material: SUS316
11-14
11.3 Options
Material: SUS316
013656 S-T union 1 mm 1
Material: SUS316
Material: PCTFE
008278 T-joint 1
1/4-28UNF screw
Material: PCTFE
013652 T-joint, stainless steel 1 set
1/4-28UNFscrew
Material: SUS316
11-15
MEMO
11-16
Chapter 12
Theory
12.1 Instrument Configuration
The general flow diagram of this instrument is shown below. The flow channel is of the
reference flow type using two pumps. To prevent performance deterioration when a
semi-micro column such as Tosohs Super H type, this instrument is designed to
minimize the dead volume between injection valve, column, and detector.
Drain block
Pump oven
12-2
12.2 Pump Unit
This pump unit is the reference flow type using two pumps as below.
Drain
(To drain block)
Drain valve
Stop valve
Vacuum Inline filter Inline filter
chamber (sample) (reference)
Solenoid
valve NC
Vacuum pump Purge pump
COM Pump Pump
NO (sample) (reference)
Inlet coil
12-3
12.3 Auto Sampler
The sampler of this instrument employs a method whereby the sample is loaded to the
sample loop connected to the injection valve, and the valve is switched to inject the
sample into the column.
<Injection operation>
With the needle connected to the syringe, wash solvent loaded to the syringe is
discharged from the needle that has moved to the washing port. (washing)
In order to prevent the sample from mixing with the wash solvent while it is being
loaded to the syringe, a volume of air equivalent to the air volume is loaded from
the end of the needle.
The needle drops into the specified sample cup, the sample is loaded to the
syringe, and the tube between the needle and the injection valve is filled with
sample.
The injection valve is switched, causing the needle, sample loop, and syringe to be
connected together, and the set injection volume of the sample is loaded to the
sample loop. (LOAD)
When the injection valve switches, the sample inside the sample loop is pushed
out by the solvent from the pump, and injected into the column. (INJECTION)
Sample loop
4way valve
Needle
Washing port
Injection valve
Pump Column
Syringe
Wash solvent
The sample loading volume is the sum of the injection volume and the presuction
volume.
12-4
12.4 Detector (RI)
The Bryce-type differential refractometer (RI) uses the phenomenon whereby a ray passing
through two media of different refractive indexes is deflected at their border.
A ray passing through the two cells containing sample and the reference filled with solvent
is deflected at the cell border because of a difference in the refractive indexes of the
solvents. The ray deflection is detected by the light-receiving element as a change
(displacement ) of ray position. Since the displacement is proportional to the difference
in refractive index between the two solvents, the differential refractive index can be
obtained by measuring the displacement .
Reference cell
Displacementment
Ray
Light receiving
element
Sample cell
12-5
12.4 Detector (RI)
A ray emitted from the tungsten lamp passes through the slit and the lens and enters the
cell. The ray deflected in the cell is reflected in the mirror and then enters the cell again.
After passing through the cell, the ray passes through the lens and the null glass and forms
a slit-shaped image on the light-receiving element. The light-receiving element consists of
two light receivers and the light-receiving quantity of the light receivers varies with changes
of the image position caused by ray deflection. A differential refractive index can be
obtained from the difference in output between the light receivers.
Tungsten lamp
Slit
Mirror
Lens
Ray
Cell
Light receiving element
Light receivers
12-6
12.5 Detector (UV)
1 Vsam
A = log (T = )
T Vref
Sample photodiode
Reference photodiode
Divided light
12-7
MEMO
12-8
Chapter 13
References
13.1 Solvent Miscibility
Before solvent replacement, check the miscibility of both solvents using the table below.
When replacing the solvent with a non-miscible solvent, substitute the solvent with a
solvent that is miscible with both solvents.
NAME
ACETIC ACID
ACETONE Not miscible
ACETONITRILE
BENZENE
Miscible
BUTYL ALCOHOL
CARBON TETRACHLORIDE
CHLOROFORM
CYCLOHEXANE
CYCLOPENTANE
DICHLOROETHANE
DICHLOROMETHANE
DIMETHYLFORMAMIDE
DIMETHYL SULFOXIDE
DIOXANE
ETHYL ACETATE
ETHYL ALCOHOL
DI-ETHYLETHER
HEPTANE
HEXANE
METHYL ALCOHOL
METHYLETHYL KETONE
1-OCTANE
PENTANE
1-PROPYL ALCOHOL
DI-PROPYLETHER
TETRACHRLOROETHANE
TETRAHYDOROFURANE
TOLUENE
TRICHLORORETHANE
WATER
XYLENE
CARBON TETRACHLORIDE
TETRACHRLOROETHANE
TETRAHYDOROFURANE
METHYLETHYL KETONE
DIMETHYLFORMAMIDE
DIMETHYL SULFOXIDE
TRICHLORORETHANE
1-PROPYL ALCOHOL
DICHLOROMETHANE
DICHLOROETHANE
METHYL ALCOHOL
DI-PROPYLETHER
BUTYL ALCOHOL
ETHYL ALCOHOL
ETHYL ACETATE
CYCLOPENTANE
DI-ETHYLETHER
CYCLOHEXANE
ACETONITRILE
CHLOROFORM
ACETIC ACID
1-OCTANE
ACETONE
HEPTANE
PENTANE
TOLUENE
BENZENE
DIOXANE
HEXANE
XYLENE
WATER
13-2
13.2 Solvent Characteristics
GPC uses some solvents that are harmful to the human body and the environment. This
section introduces some general solvent characteristics. The values may vary with the
measurement conditions.
For details, refer to the MSDS created by the solvent manufacturer.
Solvent Tetrahydrofuran
Mol. formula C4H8O Refractive index 1.4050 (20C)
Mol. mass 72.1 Fusion point -108.5C
Structural formula Boiling point 65C
Specific gravity 0.8892 (20C)
Flash point -14C
Ignition point 321C
Explosion limit 2.0 to 11.8 vol%
CAS No. 109-99-9
Solvent Chloroform
Mol. formula CHCl3 Refractive index 1.4467 (20C)
Mol. mass 119.38 Fusion point -64C
Structural formula Boiling point 62C
Specific gravity 1.484(20C)
Flash point no-data
Ignition point no-data
Explosion limit no-data
CAS No. 67-66-3
13-3
13.2 Solvent Characteristics
Solvent N,N-Dimethylformamide
Mol. formula C3H7NO Refractive index 1.4269 (25C)
Mol. mass 73.1 Fusion point -61C
Structural Boiling point 153.0C
formula Specific gravity 0.9445 (25C)
Flash point 58C
Ignition point 410C
Explosion limit 2.2 to 16 vol%
CAS No. 68-12-2
Solvent Dimethylsulfoxide
Mol. formula C2H6OS Refractive index 1.4783 (25C)
Mol. mass 78.1 Fusion point 18.42C
Structural formula Boiling point 189C
Specific gravity 1.1014 (20C)
Flash point 84C(sealed)
Ignition point 270C
Explosion limit 2.6 to 28.5 vol%
CAS No. 67-68-5
13-4
13.2 Solvent Characteristics
Solvent Toluene
Mol. formula C7H8 Refractive index 1.497 (20C)
Mol. mass 92.14 Fusion point -95C
Structural formula Boiling point 111.6C
Specific gravity 0.867 (20C)
Flash point 4 to 4.4C
Ignition point 116 to 480C
Explosion limit 1.2 to 7.1 vol% (in the
atmosphere)
CAS No. 108-88-3
Solvent N-Methyl-2-pyrrolidone
Mol. formula C5H9NO Refractive index no-data
Mol. mass 99.14 Fusion point -22.97C
Structural Boiling point 204C
formula Specific gravity 1.027 (25C)
Flash point 99C
Ignition point no-data
Explosion limit 0.9 to 3.9 % (in the air)
CAS No. 872-50-4
Solvent m-Cresol
Mol. formula C7H8O Refractive index 1.5438 (20C)
Mol. mass 108.14 Fusion point 11.95C
Structural formula Boiling point 203C
Specific gravity 1.034 (20C)
Flash point 86C
Ignition point 558.9C
Explosion limit 1.06 vol% (lower limit)
CAS No. 108-39-4
13-5
13.2 Solvent Characteristics
Acetonitrile 41.05 82 1.345 200 nm: 0.22, 210 nm: 0.10 0.782 to 0.784
220 nm: 0.10, 230 nm: 0.02
Benzene 8.11 80 1.501 280 nm: 0.70, 290 nm: 0.10 0.877 to 0.881
300 nm: 0.05
Carbon tetrachloride 153.82 77 1.460 265 nm: 0.80, 270 nm: 0.32 1.594 to 1.599
280 nm: 0.08
Chloroform 119.38 61 1.445 254 nm: 0.30, 270 nm: 0.03 1.477 to 1.494
280 to 400 nm: 0.01
Cyclohexane 84.16 81 1.426 210 nm:1.00, 220 nm: 0.50 0.777 to 0.781
230 nm: 0.20, 240 nm: 0.10
1.2-Dichloroethane 98.96 83 1.445 225 nm:1.00, 230 nm: 0.50 1.255 to 1.260
240 nm: 0.10, 254 nm: 0.02
Dichloromethane 84.93 40 1.425 220 nm: 0.07, 230 nm: 0.50 1.033 to 1.038
240 nm: 0.40, 254 nm: 0.20
Dimethylformamide 73.10 153 1.430 270 nm: 0.70, 275 nm: 0.30 0.949 to 0.954
295 nm: 0.10, 310 nm: 0.05
Dimethyl sulfoxide 78.13 -- 1.476 270 nm: 0.50, 280 nm: 0.25 1.100 to 1.102
290 nm: 0.20, 313 nm: 0.05
Dioxane 88.11 101 1.422 220 nm: 0.70, 230 nm: 0.50 1.033 to 1.038
240 nm: 0.40, 254 nm: 0.20
Ethyl acetate 80.11 77 1.373 254 nm:1.00, 280 nm: 0.02 0.899 to 0.904
260 nm: 0.30, 270 nm: 0.10
Ethyl alcohol 46.07 78 1.362 210 nm: 0.70, 240 nm: 0.10 0.790 to 0.904
220 nm: 0.40, 230 nm: 0.20
13-6
13.2 Solvent Characteristics
Methyl alcohol 32.04 65 1.329 210 nm: 0.07, 220 nm: 0.30 0.791 to 0.794
230 nm: 0.15, 240 nm: 0.07
Octane 114.23 --- 1.398 210 nm: 0.50, 240 nm: 0.02 0.702 to 0.706
220 nm: 0.15, 230 nm: 0.05
Propanol 60.10 82 1.377 210 nm: 0.70, 230 nm: 0.20 0.785 to 0.788
254 nm: 0.03, 270 nm: 0.02
Tetrahydrofurane 72.11 66 1.408 240 nm: 0.70, 254 nm: 0.20 0.887 to 0.891
280 nm: 0.10, 300 nm: 0.05
Toluene 92.14 111 1.497 285 nm:1.00, 290 nm: 0.50 0.866 to 0.869
300 nm: 0.15, 320 nm: 0.04
13-7
13.3 Quick Reference for Boiling Point
Before solvent replacement, check both boiling points using the table below. When
replacing a solvent of a high boiling point with a solvent of a low boiling point, cool the
instrument well.
When replacing a solvent of a low boiling point with a solvent of a high boiling point, do not
raise the instrument temperature until replacement is complete.
280
260 1-chloronaphthalene
240
220
1,2,4-trichlorobenzene
N-methylpyrolidone
200
N,N-Dimethylsulfoxide
o-Dichlorobenzene 180
N,N-Dimethyacetoamide
160
N,N-Dimethylformamide
140
Aceto Hexafluoroisopropanol
Heptan
40
20
13-8
13.4 Waste Liquid Treatment
This section introduces the general waste liquid treatment methods. The treatment method
may differ depending on the entrusted agent. For details, refer to the related legal
regulations.
solvent incinerated by
Oxygen-contai Alcohol, ether, spraying and the
ning solvent ketone, ester, etc. waste gas is
Nitrogen-contai Pyridine, amide, Diluted with C, H, or O washed by alkali.
ning solvent nitrile, etc. type solvent for 5 vol% 4. The amount of
Sulfur-containi Sulfide, sulfoxide, or less of N and S solvent injected
ng solvent etc. into the
combustion
chamber is
Halogen-contai Carbon tetrachloride, 5 vol% or less as a adjusted according
Incombustible waste
13-9
13.5 Countermeasures against Static Electricity
When the mobile phase is a combustible organic solvent, take great care about with regard
to fire or explosions due to static electricity.
If a liquid flows through fine tubing, as in a liquid chromatograph at a high speed, flow
electrification generates static electricity. As a charged liquid accumulates in an insulated
container, the charge level gradually increases and voltage as high as several kV is
generated. As a conductor approaches, a spark may occur and ignite ambient combustible
gas if any.
13-10
13.5 Countermeasures against Static Electricity
Ground terminal
For the easy grounding of a waste liquid, the following options are available:
Ground kit A for non-conductive waste liquid container (3-L gallon bottle)
(Part No. 0019615)
Ground kit B for conductive waste liquid container (18-L oil tubing)
(Part No. 0019616)
13-11
MEMO
13-12
Manufacturer TOSOH CORPORATION
BIOSCIENCE DIVISION
Shiba-Koen First Bldg.
3-8-2, Shiba, Minato-ku, Tokyo 105-8623, Japan
Phone: +81-3-5427-5181
Fax: +81-3-5427-5220
This manual may not be reprinted or copied in whole, or in part without written consent of TOSOH
CORPORATION. The contents of the manual are subject to change without notice.