MODEL L-2100/2130 PUMP GRADIENT EDITION

Please read this manual carefully and store it

in a safe place.

Before using the instrument, read the safety

instructions and precautions carefully.
Keep this manual in a safe place nearby so it can
be referred to whenever needed.
 INSTRUCTION MANUAL

MODEL L-2100/2130 PUMP

GRADIENT EDITION

Hitachi High-Technologies Corporation

Copyright Hitachi High-Technologies Corporation 2002. All rights reserved. Printed in Japan. Part No. 890-9106 TF-K (FT-LT)
 CONTENTS

1. OUTLINE .................................................................................................................. 1-1

1.1 Gradients................................................................. 1-1

2. INSTALLATION AND ASSEMBLY.......................................................................... 2-1

2.1 Assembly ................................................................ 2-1
2.1.1 Assembly of Low-Pressure
Gradient System .......................................... 2-1
2.1.2 Assembly of High-Pressure
Gradient System .......................................... 2-7

3. OPERATING PROCEDURES .................................................................................. 3-1

3.1 Creating an Analytical Program ............................ 3-1
3.1.1 How to Create a New Program .................... 3-3
3.1.2 How to Delete a Program............................. 3-4
3.1.3 How to Remove a Step ................................ 3-5
3.1.4 How to Add a Step ....................................... 3-6
3.2 Low-Pressure Gradient Mode................................ 3-7
3.2.1 Pump Mode Setting ..................................... 3-7
3.2.2 Low-Pressure Gradient Type Setting........... 3-8
3.2.3 Programming Operation Procedure ............. 3-9
3.2.4 Manual Operation Procedure....................... 3-10
3.2.5 Analytical Operation..................................... 3-11
3.3 High-Pressure Gradient Mode............................... 3-13
3.3.1 Pump Mode Setting ..................................... 3-13
3.3.2 Programming Operation Procedure ............. 3-16
3.3.3 Manual Operation Procedure....................... 3-17
3.3.4 Analytical Operation..................................... 3-17
3.4 Utilities .................................................................... 3-18
3.4.1 Pump Motor Control Setting......................... 3-18
3.4.2 Link Program Function................................. 3-21
3.4.3 Column Flush Setting................................... 3-30
3.4.4 Event Code Setting ...................................... 3-34

4. PERFORMANCE CHECK........................................................................................ 4-1

4.1 Checking the Mixing Ratio Accuracy in
a Gradient System .................................................. 4-1
4.1.1 Preparation Before Measurement................ 4-2
4.1.2 Conditions .................................................... 4-4
4.1.3 Mixing Ratio Calculation Method ................. 4-5
4.1.4 Specifications............................................... 4-5

-i-
5. MAINTENANCE AND CHECKUP............................................................................ 5-1
5.1 Error Messages and Countermeasures................ 5-1
5.2 Troubleshooting ..................................................... 5-3
5.3 Periodic Maintenance and Checkup ..................... 5-4
5.4 Gradient Mixer ........................................................ 5-5
5.4.1 Conventional Mixer ...................................... 5-5

6. SPARE PARTS ........................................................................................................ 6-1

6.1 Gradient Spare Parts for L-2100............................ 6-1

7. SPECIFICATIONS.................................................................................................... 7-1
7.1 Model L-2100 Low-Pressure Gradient System .... 7-1
7.2 Model L-2100 High-Pressure Gradient System.... 7-3
7.3 Model L-2130 Low-Pressure Gradient System .... 7-5
7.4 Model L-2130 High-Pressure Gradient System.... 7-7

- ii -
 1.1

1. OUTLINE

This is the gradient edition of the instruction manual for the

L-2100/2130 pump. For details on the pump itself, refer to the
pump edition of the instruction manual.
The following three kinds of gradients are available.
(1) Low-pressure gradient
(2) High-pressure gradient

1.1 Gradients

(1) Low-Pressure Gradient

Mobile phases are joined on the low pressure side of the

pump. We refer to this configuration as a low pressure
gradient mode since the mobile phases are mixed before
pressurization. The proportioning valve selects from up to
four mobile phases.

(2) High-Pressure Gradient

In this configuration, the mobile phases are mixed after

being pressurized by the pump. Accordingly this mixing
method is called a high pressure gradient mode.
This configuration uses two pumps for pressurizing the two
mobile phases independently.

1-1
 2.1.1

2. INSTALLATION AND ASSEMBLY

2.1 Assembly

2.1.1 Assembly of The piping arrangement for the low-pressure gradient system is
Low-Pressure shown below.
Gradient System

WARNING
Ignition of Flammable Chemicals!

Beware of ignition hazard when using flammable

chemicals such as organic solvents.
Always check the following conditions. If an abnormality
is found, stop operation immediately.
Leakage of solvent or waste solution
Leakage of solvent inside the instrument
Inadequate ventilation of the laboratory room
This instrument is not explosion-proof.
Although aqueous solvents or organic solvents having an
ignition point of 70 C or higher are usable, do not use
organic solvents having an ignition point below 70 C.
When using flammable chemicals, be careful about
possible ignition due to static electricity. Particularly
when using non-conductive chemicals, employ a
conductive vessel made of metal or the like and provide
grounding connection correctly.

WARNING
Explosion of Vapor from Flammable Chemicals!

If a flammable chemical such as organic solvent leaks

from the flow path of the instrument and its vapor
concentration exceeds the explosion limit, it may cause
spontaneous combustion with dangerously explosive
results.
When using flammable and readily volatile chemical, be
sure to check for leakage from the instrument flow path
and ventilate the laboratory room adequately.

2-1
2.1 Assembly

WARNING
Beware of Electric Shock!

Potentially Dangerous Voltages are Present within the

Instrument.
Before removing the instrument cover for replacement or
adjustment of inside parts, be sure to turn OFF the power
switch and unplug the power cord.

In the low-pressure gradient system, tubing must be made

with the proportioning valve attached in place.
Cautions on tubing and wiring for the system are given
below.

(a) Attachment of Proportioning Valve

1) Detach the front door. (Open the door, and lift

the bottom of the hinge part while pulling forward
to remove it.)

Front panel

2) Detach the front and top covers of the pump by

removing the screws. The two detached covers
will be used when configuring a high-pressure
gradient system, so keep them in a safe place.

2-2
 2.1.1

3) Connect the cord of the proportioning valve to the

connector of the pump main body.

Connector

4) Mount the proportioning valve on the pump main

body, and secure it with two screws.

Screws Tubes

2-3
2.1 Assembly

5) Take the four-way joint of tubing that extends from

the solenoid valve and insert it into the IN side of
the pump head.

Four-way joint

6) When mounting the four-way joint to the pump

head IN side, hold the joint in place with a finger
and turn the outer ring. The tube can thus be
easily attached (and detached) without twisting.

7) Connect the inlet tubes to the IN side of each

solenoid valve.

8) Reattach the covers to the pump main body.

2-4
 2.1.1

(b) System Tubing Connections

After attaching the proportioning valve unit, connect

the tubes as shown in Fig. 2-1.

0.33TT

0.25TT

Colum
0.8ST
Detector
1 Drain
Flow cell unit 2

6 3 0.25ST

0.25ST 5 4 Sample injector

Pump
Drain valve Sample drain
bottle

c Mixer
Drain
D
Pump head
C

B
A

a b

Mobile phase container Solvent filter

Fig. 2-1 Tubing of Low-Pressure Gradient System

2-5
2.1 Assembly

If air bubbles are contained in the tubing between

mobile phase container and detector outlet in the low-
pressure gradient system, full performance cannot be
expected from the system. Before using this system,
therefore, make sure that no air is introduced through
the tubing joints. Take the following procedure for
this check.

1) Connect the tubes, open the drain valve, then

conduct purging by feeding each mobile phase
through the tubing.

2) Check for air bubbles

(i) With each tube being filled with mobile phase,
open the drain valve and perform purging with
liquid A. At this step, check that there are no
air bubbles at part [a].
(ii) In the same way, perform purging with liquids
B, C and D, and check that there are no air
bubbles at part [a].
(iii) If air bubbles remain after repeating the
above steps (i) and (ii) a few times, retighten
by hand the setscrew of the solenoid valve at
part [b].
(iv) While continuing the purge, check that no air
bubbles appear at part [c].
(v) Check that no air is introduced during liquid
feed (or purging).

3) Stabilization
Make sure that no air is introduced.

2-6
 2.1.2

2.1.2 Assembly of The high-pressure gradient system is to be installed in front of

High-Pressure the high-pressure gradient unit pump. Cautions on wiring and
Gradient System tubing of the system are given below.

(1) Wiring

WARNING
Ground properly to prevent an electric shock hazard!

Be sure to use the power cable supplied with the

instrument. Use of a different power cable could result in
an electric shock hazard.
This instrument is of a plug-connected type specified in
European Norm EN61010-1, so connect the power cable
to a grounded three-wire outlet.
If a grounded three-wire outlet is not available, the power
cable may be connected to a two-wire outlet through an
adapter plug. In this case, be sure to provide proper
grounding connection as required.

The wiring for the high-pressure gradient system is

described next. An e-Line is used for connection between
pumps. Connect one end of the e-Line cable to the
upstream side of e-Line connector of pump A, and the other
end of the cable to the downstream side of the e-Line
connector of pump B.

(2) e-Line Setting for High-Pressure Gradient System

(a) For the high-pressure gradient system, be sure to

connect the e-Line cable directly between the pumps.
If connected via another module, control of the high-
pressure gradient system would be disabled.

(b) If CDS is not used in the high-pressure gradient

system, set the pump Control Mode to Internal.

NOTE: Setting the pump Control Mode to External here

would disable the control.

2-7
2.1 Assembly

(3) Tubing

WARNING
Ignition of flammable chemicals!

Beware of ignition hazard when using flammable

chemicals such as organic solvents.
Always check the following conditions. If an abnormality
is found, stop operation immediately.
Leakage of solvent or waste solution
Leakage of solvent inside the instrument
Inadequate ventilation of the laboratory room
This instrument is not explosion-proof.
Although aqueous solvents or organic solvents having an
ignition point of 70 C or higher are usable, do not use
organic solvents having an ignition point below 70 C.
When using flammable chemicals, be careful about
possible ignition due to static electricity.
Particularly when using non-conductive chemicals, employ
a conductive vessel made of metal or the like and provide
grounding connection correctly.

WARNING
Explosion of vapor from flammable chemicals!

If a flammable chemical such as organic solvent leaks

from the flow path of the instrument and its vapor
concentration exceeds the explosion limit, it may cause
spontaneous combustion with dangerously explosive
results.
When using a flammable and readily volatile chemical, be
sure to check for leakage from the instrument flow path
and ventilate the laboratory room adequately.

2-8
 2.1.2

WARNING
Beware of electric shock!

Potentially dangerous voltages are present within the

instrument.
Before removing the instrument cover for replacement or
adjustment of inside parts, be sure to turn OFF the power
switch and unplug the power cord.

(a) The components of the high-pressure gradient system

are to be mounted to the front of the pump.
The mounting procedure is explained next

1) Remove the round plate on the front of pump by

loosening two screws. Then detach the tubing
between pumps A and B.

Screws

2-9
2.1 Assembly

2) Attach the high-pressure unit using the two

furnished screws. Connect wiring to the
furnished high-pressure unit circuit board and
main unit connector.

Connectors Screws

2 - 10
 2.1.2

3) Detach the discharge valve from the pump head,

and attach it to the upper part of the high-pressure
unit.

Filter holder Discharge valve

Attach the furnished filter and filter holder to the

pump head. Then connect the two furnished
tubes.

4) Attach the high-pressure unit circuit board to the

pump main unit.
Screws

2 - 11
2.1 Assembly

(b) Tubing of System

After mounting the high-pressure unit, carry out tubing

as shown below.

0.33TT

0.25TT

Column
0.8ST
Detector
1 2 Drain
Flow cell unit

6 3 0.25ST

0.25TT 5 4 Sample injector

Pump
Sample drain
Drain valve bottle
0.25ST

Mixer
Drain Pump head

0.25ST

Pump

Drain valve

Drain Pump head

1.5TT

Mobile phase container Solvent filter

Fig. 2-2 Tubing of High-Pressure Gradient System

2 - 12
 3.1

3. OPERATING PROCEDURES

3.1 Creating an Analytical Program

(1) Up to nine set programs can be created. Using the SET

PROGRAM function, select or prepare a desired program.
For gradient analysis, take the following steps:

(a) Press the PROG key, and select or prepare an elution

condition program.

(b) Turn on the pump.

(c) Start the program.

(2) SET PROGRAM Function

Using the SET PROGRAM function, specify the following

items; an upper pressure limit, lower pressure limit, mixing
ratio, flow rate, and event.

(a) %C and %D are not indicated in the two-liquid high-

pressure gradient mode.

(b) In the EVENT field, up to four event codes can be set

up per TIME. Thus, you can specify conditions of
contact signal output to the EVENT terminals on the
rear panel for internal operations (stop of pump,
sounding of buzzer).

(c) When it is desired to set up just a particular item such

as EVENT, press the ENT key for %B, %C, %D, or
FLOW.

3-1
3.1 Creating an Analytical Program

When creating a time program using the SET

PROGRAM function, provide interpolation for a mixing
ratio and flow rate as shown below.

Parameter Mixing Ratio Flow Rate

Interpolation
Linear Linear

TN TN+1 TN TN+1

TN, TN+1: Preset time

NOTICE: For creating a stepwise program, apply TN+1 - TN

= 0.1 (min).

3-2
 3.1.1

3.1.1 How to Create a Demonstrated below is how to create a new program.

New Program

PROG
* On the subsequent screens, press the ENT key
after input of a numeric value.

SET PROGRAM NO. (1 - 9)

1

Enter a number corresponding to a new program

(enter a numeric value ranging from 1 to 9).

EDIT MODE (MODIFY = 0, NEW = 1)

0 1

Specify whether program modification is to be made

(MODIFY = 0) or new program creation is to be made
(NEW = 1).

PRESSURE (MAX MIN)

< 0.0 - 39.2 > 39.2 0.0 1

Enter a value of the upper pressure limit (MAX) and a

value of the lower pressure limit (MIN).

TIME B C D FLOW EVENT

0.0 0 0 0 1.000 1

Enter a time period (TIME), mixing percentage of

solution B (%B), mixing percentage of solution C (%C),
mixing percentage of solution D (%D), flow rate
(FLOW), and event number (EVENT).

TIME B C D FLOW EVENT

1

When a step subsequent to the end of program input

is reached on the screen, press the ESC key.

MONITOR
screen

3-3
3.1 Creating an Analytical Program

3.1.2 How to Delete a Demonstrated below is how to delete a program.

Program
PROG

Press the PROG key.

SET PROGRAM NO. (1 - 9)

1

Enter a number corresponding the program to be

deleted.

EDIT MODE (MODIFY = 0, NEW = 1)

0 1

Enter 1 .
The specified existing program will be deleted.

ESC Pressing the ESC key brings control back to the

MONITOR screen.

3-4
 3.1.3

3.1.3 How to Remove a Demonstrated below is how to remove a step from a program.
Step

PROG

SET PROGRAM NO. (1 - 9)

1

Enter a number corresponding to the program

concerned.

PRESSURE (MAX MIN)

< 0.0 - 39.2 > 39.2 0.0 1
Enter values of the upper and lower pressure limits.

TIME B C D FLOW EVENT

0.0 0 0 0 1.000 1

Using the key, go to the step to be removed.

If you go beyond the step to be removed, return to it
using the key.
With the cursor located at TIME, press the DEL key.

TIME B C D FLOW EVENT

1

Pressing the ESC key terminates the current session.

Thus, the MONITOR screen appears again.

ESC

3-5
3.1 Creating an Analytical Program

3.1.4 How to Add a Demonstrated below is how to add a step in a program.

Step
PROG

SET PROGRAM NO. (1 - 9)

1

Enter a number corresponding to the program

concerned.

PRESSURE (MAX MIN)

< 0.0 - 39.2 > 39.2 0.0 1

Enter values of the upper and lower pressure limits.

TIME B C D FLOW EVENT

0.0 0 0 0 1.000 1

Using the or key, go to a position

corresponding to the new step to be added.
When you reach it, enter desired parameters;
TIME, %B, %C, %D, FLOW and EVENT.

TIME B C D FLOW EVENT

1

Pressing the ESC key terminates the current session.

ESC
Thus, the MONITOR screen appears again.

3-6
 3.2.1

3.2 Low-Pressure Gradient Mode

The following describes how to operate the L-2100/2130 pump in

the low-pressure gradient mode.

3.2.1 Pump Mode

NOTICE: Since the pump mode setting is retained in
Setting
internal memory once it is made, it is not required
to make the pump mode setting for each
operation.

UTLTY
POWER ON + (SETUP)
UTLTY
Turn power on while holding down the (SETUP) key.
UTLTY
The SETUP screen will then appear. Note that the (SETUP)
key should be held down until the SETUP screen comes up.

SETUP: PUMP MODE CONTRAST S.NO.

Micro A LOW 4 02AB-111

Press the ENT key.

(Regular will be indicated in the case of the L-2130
pump.)

SETUP ITEMS (PUMP = 1, CONTRAST = 2)

1

Press the 1 key, followed by the ENT key.

PUMP (A = 1, B = 2)
1

Enter a pump station number.

Press the 1 key, followed by the ENT key.

GR MODE (LOW =1, HIGH = 2, V. HIGH = 3,

1 FLOW = 4)

Select the low-pressure gradient mode for the pump.

LOW : Low-pressure gradient mode
HIGH : High-pressure gradient mode
V.HIGH : (not used)
FLOW : Isocratic mode

SETUP: PUMP MODE CONTRAST S.NO.

Micro A LOW 4 02AB-111

3-7
3.2 Low-Pressure Gradient Mode

POWER OFF

Turn power off once.

POWER ON

Turn power on again.

Self-diagnosis will be carried out.

TIME A B C D FLOW PRESS [--]

0.0 100 0 0 0 0.000 0.0 1

After the INITIAL screen appears, the MONITOR

screen in the low-pressure gradient mode is
presented.

3.2.2 Low-Pressure In the low-pressure gradient system, specify the kind of mixer,
Gradient Type requirement/non-requirement of degasser, type of gradient
Setting operation according to the kind of solvent and flow rate to set up
an instrumental arrangement. In general, a mixer having a
larger capacity is required as the flow rate increases.
Furthermore, it is advisable to prepare a large-capacity mixer for
mixing sparingly mixable solvents. Use a degasser to enhance
accuracy in mixing ratio for ensuring good reproducibility in
analysis. The following table shows examples of instrumental
arrangements for mixing water and methanol.

Flow Rate Range

(mL/min)
Without mixer
Semi-micro mixer 0.2 to 0.4
Conventional mixer 0.4 to 1.8

* The allowable flow rate range per flow path in the L-2100/
2130 degasser is 3.0 mL/min or less.

3-8
 3.2.3

3.2.3 Programming Demonstrated below is a program-based operational procedure

Operation in the low-pressure gradient mode.
Procedure
MONITOR
screen

If the MANUAL screen is selected, press the INIT key

for changeover to the programming mode screen.

PROG

Press the PROG key.

SET PROGRAM NO. (1 - 9)

1

Enter a program number in a range of 1 to 9.

EDIT MODE (MODIFY = 0, NEW = 1)

0 1
Specify whether program modification is to be made
(MODIFY = 0) or new program creation is to be made
(NEW = 1).

PRESSURE (MAX MIN)

< 0.0 - 39.2 > 39.2 0.0 1

Enter a value of the upper pressure limit (MAX) and a

value of the lower pressure limit (MIN).

TIME B C D FLOW EVENT

0.0 0 0 0 1.000 1

Enter a time period (TIME), mixing percentage of

solution B (%B), mixing percentage of solution C (%C),
mixing percentage of solution D (%D), flow rate
(FLOW), and event number (EVENT).
TIME B C D FLOW EVENT
1

When a new input screen appears, enter a time

period (TIME), mixing percentage of solution B (%B),
mixing percentage of solution C (%C), mixing
percentage of solution D (%D), flow rate (FLOW), and
event number (EVENT). Upon completion of this
input step, go to another new input screen.
ESC

Press the ESC key to return to the

MONITOR screen.
MONITOR
screen

3-9
3.2 Low-Pressure Gradient Mode

NOTICE: 1. Up to 99 steps can be input in the total of step

counts in programs No.1 to 9.
2. An input program is retained in internal
memory. When power is turned on next time,
the initial state screen (TIME = 0) of a program
used immediately before power-off is
presented.

3.2.4 Manual Operation Demonstrated below is a manual operation procedure in the low-
Procedure pressure gradient mode.
Use manual operation for liquid feeding under a certain condition
without using a program.

MONITOR
screen

Press the 0 key, and the MANUAL mode screen

will then appear.

MANUAL ( B C D FLOW MAX MIN )

0 0 0 1.000 39.2 0.0

MANUAL A B C D FLOW PRESS [MN]

90 10 0 0 1.000 0.0 1

MONITOR
screen

NOTICE: Manual operation conditions are deleted when

power is turned off. Set up manual operation
conditions each time after power-on.

3 - 10
 3.2.5

3.2.5 Analytical
Operation WARNING
Ignition of Flammable Chemicals!

Beware of ignition hazard when using flammable

chemicals such as organic solvents.
Always check the following conditions. If an abnormality
is found, stop operation immediately.
Leakage of solvent or waste solution.
Leakage of solvent inside the instrument.
Well-ventilate the laboratory room where the
instrument is used.
This instrument is not explosion-proof.
Although aqueous solvents or organic solvents having an
ignition point of 70 C or higher are usable, do not use
organic solvents having an ignition point below 70 C.
When using flammable chemicals, be careful about
possible ignition due to static electricity.
Particularly when using non-conductive chemicals, employ
a conductive vessel and provide grounding connection
correctly.

WARNING
Explosion of Vapor from Flammable Chemicals!

If a flammable chemical such as organic solvent leaks

from the flow path of the instrument and its vapor
concentration exceeds the explosion limit, it may cause
spontaneous combustion with dangerously explosive
results.
When using a flammable and readily volatile chemical, be
sure to check for leakage from the instrument flow path
and ventilate the laboratory room adequately.

3 - 11
3.2 Low-Pressure Gradient Mode

After completion of preparatory setting, take the following

procedure for analytical operation.

(1) Check that the pump, column, detector and data processor
are connected properly.

(2) Set the mobile phase to be used and the waste solution
bottle.

(3) Start up the pump, and wait until the flow path is filled with
the mobile phase.

(4) Wait until completion of the warming-up of the pump,

column and detector, i.e., wait until the baseline becomes
stable.

(5) Inject the sample to be examined.

(6) After completion of analysis, turn power off and treat waste
solution properly.

3 - 12
 3.3.1

3.3 High-Pressure Gradient Mode

The following describes how to operate the L-2100/2130 pump in

the high-pressure gradient mode.

3.3.1 Pump Mode In the high-pressure gradient mode, set up liquid feed conditions
Setting for each of PUMP A and PUMP B.

NOTICE: Since the pump mode setting is retained in

internal memory once it is made, it is not required
to make the pump mode setting for each
operation.

(1) For PUMP A

UTLTY
POWER ON + (SETUP)
UTLTY
(SETUP)
Turn power on while holding down the key. UTLTY
The SETUP screen will then appear. Note that the (SETUP)
key should be held down until the SETUP screen comes up.

SETUP: PUMP MODE CONTRAST S.NO.

Micro A LOW 4 02AB-111

Press the ENT key.

(Regular will be indicated in the case of the L-2130
pump.)

SETUP ITEMS (PUMP = 1, CONTRAST = 2)

1

Press the 1 key, followed by the ENT key.

PUMP (A = 1, B = 2)
1

Enter a pump station number.

Press the 1 key, followed by the ENT key.

GR MODE (LOW = 1, HIGH = 2, V. HIGH = 3

1 FLOW = 4)

Select the high-pressure gradient mode for the pump.

LOW : Low-pressure gradient mode
HIGH : High-pressure gradient mode
V.HIGH : (not used)
FLOW : Isocratic mode

3 - 13
3.3 High-Pressure Gradient Mode

CONTROL MODE (EXTERNAL = 1, INTERNAL = 2)

1

Select INTERNAL for the CONTROL MODE.

Use EXTERNAL mode where control is conducted by
the personal computer.

SETUP: PUMP MODE CONTRAST S.NO.

Micro A HIGH (INT) 4 02AB-111

POWER OFF

Turn power off once.

POWER ON

Turn power on again.

Self-diagnosis will be carried out.

TIME A B FLOW PRESS [--]

0.0 100.0 0.0 0.000 0.0 1
After the INITIAL screen appears, the MONITOR
screen in the high-pressure gradient mode is
presented.

(2) For PUMP B

UTLTY
POWER ON + (SETUP)

UTLTY
(SETUP)
Turn power on while holding down the key.
UTLTY
The SETUP screen will then appear. Note that the (SETUP)
key should be held down until the SETUP screen comes up.

3 - 14
 3.3.1

SETUP: PUMP MODE CONTRAST S.NO.

Micro A LOW 4 02AB-111

Press ENT key.

(Regular will be indicated in the case of the L-2130
pump.)

SETUP ITEMS (PUMP = 1, CONTRAST = 2)

1

Press the 1 key, followed by the ENT key.

PUMP (A = 1, B = 2)
2

Enter a pump station number.

Press the 2 key, followed by the ENT key.

GR MODE (LOW = 1, HIGH = 2, V. HIGH = 3,

2 FLOW = 4)

Select the high-pressure gradient mode for the pump.

LOW : Low-pressure gradient mode
HIGH : High-pressure gradient mode
V.HIGH : (not used)
FLOW : Isocratic mode

CONTROL MODE (EXTERNAL = 1, INTERNAL = 2)

2

Select INTERNAL for the CONTROL MODE.

NOTE: Control will be disabled if EXTERNAL is
selected.
SETUP: PUMP MODE CONTRAST S.NO.
Micro HIGH (INT) 4 02AB-111

POWER OFF

Turn power off once.

POWER ON

Turn power on again.

Self-diagnosis will be carried out.

PUMP B [- -]

After the INITIAL screen appears, the MONITOR

screen in the high-pressure gradient mode is
presented for PUMP B.

3 - 15
3.3 High-Pressure Gradient Mode

3.3.2 Programming Demonstrated below is a program-based operational procedure

Operation in the high-pressure gradient mode.
Procedure
MONITOR
screen

If the MANUAL screen is selected, press the INIT key

for changeover to the programming mode screen.

PROG

Press the PROG key.

SET PROGRAM NO. (1 - 9)

1

Enter a program number in a range of 1 to 9.

EDIT MODE (MODIFY = 0, NEW = 1)

0 1

Specify whether program modification is to be made

(MODIFY = 0) or new program creation is to be made
(NEW = 1).

PRESSURE (MAX MIN)

< 0.0 - 39.2 > 39.2 0.0 1

Enter a value of the upper pressure limit (MAX) and a

value of the lower pressure limit (MIN).

TIME B FLOW PRESS [- -]

0.0 0.0 1.000 0.0 1

Enter a time period (TIME), mixing percentage of

solution B (%B), flow rate (FLOW), and event number
(EVENT).

TIME B FLOW EVENT

1

When a new input screen appears, enter a time

period (TIME), mixing percentage of solution B (%B),
flow rate (FLOW), and event number (EVENT).
Upon completion of this input step, go to another new
input screen.

ESC

Press the ESC key to return to the MONITOR screen.

MONITOR
screen

3 - 16
 3.3.3

NOTICE: 1. Up to 99 steps can be input in the total of step

counts in programs No.1 to 9.
2. An input program is retained in internal
memory. When power is turned on next time,
the initial state screen (TIME = 0) of a
program used immediately before power-off is
presented.

3.3.3 Manual Operation Demonstrated below is a manual operation procedure in the

Procedure high-pressure gradient mode.
Use manual operation for liquid feeding under a certain condition
without using a program.

MONITOR
screen

Press the 0 key, and the MANUAL mode screen

will then appear.

MANUAL (B FLOW MAX MIN)

0.0 1.000 39.2 0.0

Enter values of mixing percentage of solution B, flow

rate, upper pressure limit, and lower pressure limit.

MONITOR
screen

NOTICE: Manual operation conditions are deleted when

power is turned off. Set up manual operation
conditions each time after power-on.

3.3.4 Analytical Refer to section 3.2.5.

Operation

3 - 17
3.4 Utilities

3.4 Utilities

3.4.1 Pump Motor In the high-pressure gradient mode, you can make pump motor
Control Setting control (M. CONTROL) selection.
In common practice, select STANDARD for M. CONTROL.
When the STANDARD mode is selected in which automatic flow
control is performed, pump-to-pump pressure may be affected
under a certain operating condition (high-pressure gradient,
reaction pump), causing an increase in baseline fluctuation.
In such a case, it is advisable to use the FIX mode in which
invariable flow rate control is performed for reducing the adverse
effect. Note, however, that the FIX mode may provide poor
baseline stability in an application where significant pressure
fluctuation occurs due to variation in mixing ratio. In this case, it
is recommended to select the STANDARD mode.

NOTICE: After checking baseline stability under practical

analysis conditions, select the STANDARD mode
or FIX mode.

Figure 3-1 shows the relationship between applicable pressure

and FIX STEP AREA in cases where baseline fluctuation is
minimized in the use of a water-soluble or organic solvent.

300

Organic solvent
200

100

Aqueous solvent

0
0 9.8 19.6 29.4 39.2

Fig. 3-1 Relationship between Pressure and

FIX STEP AREA

3 - 18
 3.4.1

(1) Using the FIX Mode

(a) Adjustment Procedure

Enter a value of FIX STEP AREA determined from the

graph shown in Fig. 3-1.

(b) Fine Adjustment Procedure

1) Select FIX STEP for pump A, and enter a value of

FIX STEP AREA determined from the graph
shown in Fig. 3-1.
2) For pump B, set M. CONTROL to STANDARD.
3) Perform liquid feeding under actual analytical
conditions (mixing ratio, flow rate, column, etc.) to
let the data processor draw a baseline.
Check fluctuation on the baseline thus drawn.
4) While varying FIX STEP AREA to an extent of 5 to
10, determine a value at which baseline
fluctuation is minimized.
5) For another pump, set M. CONTROL to
STANDARD also, and determine a value of FIX
STEP AREA at which baseline fluctuation is
minimized.
6) After a value of FIX STEP AREA is determined for
each pump, set M. CONTROL to FIX for all the
pumps. Then, enter the value of FIX STEP
AREA for each pump.

3 - 19
3.4 Utilities

(2) FIX Mode Setting Procedure

UTLTY

UTILITY (CONDITION = 1, UNIT = 2,

1 OTHERS = 3)

Select CONDITION.
Press the 1 key, followed by the ENT key.

CONDITION SET (Z-ADJ = 1, MOTOR-CTRL = 2,

2 CALIBRATION = 3)

Select MOTOR-CONTROL.
Press the 2 key, followed by the ENT key.

MOTOR CONTROL (STANDARD = 1, FIX = 2)

2

Select FIX (fixed flow rate control).

Press the 2 key, followed by the ENT key.

FIX STEP AREA (0, 30 - 300)

100

Enter a determined numeric value.

(e.g., Press 1 0 0 ENT.)

MONITOR
screen

3 - 20
 3.4.2

3.4.2 Link Program This function is available for linking two or more programs.
Function
NOTICE: For connection with Ez chrom Elite for Hitachi,
specify NO. This setting can be made on the
System Manager.

(1) Link Program Setting

Define link program conditions. The following presents an

example in which one execution cycle of program No.1 and
a conditioning time of 30 minutes are set at step 1 and
three execution cycles of program No. 2 and a conditioning
time of 20 minutes are set at step 2.

MONITOR
screen

UTLTY
(SETUP)

UTILITY (CONDITION = 1, UNIT = 2, FLUSH = 3

5 EVENT = 4, LINK = 5, OTHERS = 6)

Select LINK (LINK PROGRAM).

LINK PROGRAM (YES = 1, NO = 0)

1

Select YES.

LINK RESTART (YES = 1, NO = 0)

0

1 ENT
MONITOR
screen

After some steps of a link program are carried out,

control is returned to the first step instead of executing
the subsequent steps. (In an instance where a link
program having five steps is selected and the start of
link is specified after the end of step 2, execution
control is returned to step 1 instead of carrying out
steps 3 and subsequent.)
In a case where the same step is selected for multiple
cycles of execution and the start of link is specified
before completion of all the cycles, execution control
is returned to the first step of the link program.

3 - 21
3.4 Utilities

It is allowed to make setting under the condition that

the pump is in the initial status (the status indication
[IN] is given at the upper right of the LCD).
You cannot make setting while gradient operation or
conditioning is in execution.

0 ENT

STEPS PROG-NO. CYCLE CONDITION TIME

1 1 1 0.0

Enter a program number (PROG-NO.), a repeat count

(CYCLE), and a conditioning time (COND. TIME).

STEPS PROG-NO. CYCLE CONDITION TIME

2 1 1 0.0

Proceed to condition setting for second step.

Enter a program number (PROG-NO.), a repeat count
(CYCLE), and a conditioning time (COND. TIME).
Input link conditions corresponding to the number of
steps to be carried out. Up to nine steps are allowed.

ESC
(LOCK OFF)

MONITOR
screen

The items on the link condition input screen are as follows:

Item Description
STEPS It is not permitted to input an execution step
number. Step numbers are counted successively.
Up to nine steps are allowed.
PROG - NO. Enter a code number of the program to be linked.
The allowable input range is 1 to 9.
CYCLE Enter a repeat count of the program concerned.
The allowable input range is 1 to 99.
COND. TIME Enter a conditioning time for the execution step
concerned. The allowable input range is 0.0 to
600.0 (min). During the conditioning time, liquid
feeding is performed under the initial condition
(TIME = 0.0) of the execution step. After the
lapse of the conditioning time, program execution is
started upon input of the start signal.

3 - 22
 3.4.2

NOTICE: Where the same step is repeated (CYCLE 2 or

more), a conditioning time is not taken for
execution at the time of transition from the
preceding cycle to the subsequent cycle.
A column conditioning time should be specified
for analysis in which the same step is repeated.
Provide a sufficient length of conditioning time
since it has an effect on reproducibility of a
retention time.
For a conditioning time for repetition of the same
step, enter a conditioning program at the end of a
program specified for the program concerned.

(2) Link Program Operation

For link program execution, two or more programs are used

in succession. Demonstrated below is an example of link
program setting according to particular conditions.

i. Programs

PROGRAM No. = 1

PRESSURE VALUE : MAX MIN MPa

19.6 1.0

TIME CONCENTRATION FLOW EVENT

(min) %A %B %C %D (ml/min) 1 2 3 4
0.0 10 90 0 0 1.000
5.0 10 90 0 0
10.0 20 80 0 0
15.0 20 80 0 0
20.0 10 90 0 0

PROGRAM No. = 2

PRESSURE VALUE : MAX MIN MPa

19.6 1.0

TIME CONCENTRATION FLOW EVENT

(min) %A %B %C %D (ml/min) 1 2 3 4
0.0 30 70 0 0 1.000
5.0 40 60 0 0
10.0 40 60 0 0
15.0 30 70 0 0

3 - 23
3.4 Utilities

ii. Column Flushing Conditions

DELAY TIME = 10.0 min

COLUMN FLUSH = YES
A% = 10 %
B% = 90 %
C% = 0 %
D% = 0 %
FLOW = 6.00 ml/min
TIME = 10.0 min

iii. Link Program Conditions

STEP PROG-No. CYCLE COND.TIME (min)

1 1 1 10.0
2 2 1 10.0

(a) Start Input after Gradient Operation

In the example shown above, screen transition is made

as follows.
Each numeral enclosed in a circle in the above
example corresponds to each of the following screen
step numbers.

3 - 24
 3.4.2

Initial
TIME %A %B %C %D FLOW PRESS [IN]
0.0 10 90 0 0 1.000 5.5 1

Liquid feeding is performed in the initial status (under

the initial conditions of program No. 1 in this example).

Gradient
TIME %A %B %C %D FLOW PRESS [GR]
10.0 20 80 0 0 1.000 5.5 1

Gradient operation (execution of program No. 1 in this

example) is performed.

Conditioning time
TIME %A %B %C %D FLOW PRESS [LC]
0.0 30 70 0 0 1.000 5.5 2

During the conditioning time, liquid feeding is

performed under the initial conditions of the execution
step (program No. 2 in this example).

Initial
TIME %A %B %C %D FLOW PRESS [IN]
0.0 30 70 0 0 1.000 5.5 2

Liquid feeding is performed in the initial status (under

the initial conditions of program No. 2 in this example).

Gradient
TIME %A %B %C %D FLOW PRESS [GR]
10.0 40 60 0 0 1.000 5.5 2

When the start is input in the initial status, the

execution step (program No. 2 in this example) is
made active.

Conditioning time
TIME %A %B %C %D FLOW PRESS [LC]
0.0 10 90 0 0 1.000 5.5 1

After completion of the last step of the link program,

control is returned to the first step (program No. 1 in
this example) for execution of conditioning.

3 - 25
3.4 Utilities

Delay time (initial)

TIME %A %B %C %D FLOW PRESS [IN]
0.0 10 90 0 0 1.000 5.5 1

Liquid feeding is performed in the initial status of the

first step (program No. 1 in this example).
If the start is not input during the delay time, control
goes to column flushing ).
If the start is input during the delay time, control is
returned to ).

Column flushing
TIME %A %B %C %D FLOW PRESS [FL]
0.0 10 90 0 0 1.000 5.5 1

(b) No Start Input after Gradient Operation

In the example shown above, screen transition is made

as follows.
Each numeral enclosed in a circle in the above
example corresponds to each of the following screen
step numbers.

3 - 26
 3.4.2

Initial

TIME %A %B %C %D FLOW PRESS [IN]

0.0 10 90 0 0 1.000 5.5 1

Liquid feeding is performed in the initial status (under

the initial conditions of program No. 1 in this example).

Gradient
TIME %A %B %C %D FLOW PRESS [GR]
10.0 20 80 0 0 1.000 5.5 1

Gradient operation (execution of program No. 1 in this

example) is performed.

Conditioning time

TIME %A %B %C %D FLOW PRESS [LC]

0.0 30 70 0 0 1.000 5.5 2
During the conditioning time, liquid feeding is
performed under the initial conditions of the execution
step (program No. 2 in this example).

Delay time
TIME %A %B %C %D FLOW PRESS [IN]
0.0 30 70 0 0 1.000 5.5 2

Liquid feeding is performed in the initial status of the

first step (program No. 2 in this example).
If the start is not input during the delay time, control
goes to column flushing.

Column flushing
TIME %A %B %C %D FLOW PRESS [FL]
0.0 10 90 0 0 1.000 5.5 2

3 - 27
3.4 Utilities

(c) Stop during Execution of Gradient Operation

Initial
TIME %A %B %C %D FLOW PRESS [IN]
0.0 10 90 0 0 1.000 5.5 1

Liquid feeding is performed in the initial status (under

the initial conditions of program No. 1 in this example).

Gradient
TIME %A %B %C %D FLOW PRESS [GR]
10.0 20 80 0 0 1.000 5.5 1

Gradient operation (execution of program No. 1 in this

example) is performed.

Initial
TIME %A %B %C %D FLOW PRESS [LC]
0.0 10 90 0 0 1.000 5.5 2

Liquid feeding is performed in the initial status (under

the initial conditions of program No. 1 in this example).

Gradient
TIME %A %B %C %D FLOW PRESS [IN]
10.0 20 80 0 0 1.000 5.5 1

Gradient operation (execution of program No. 1 in this

example) is performed.
In this case, the step stopped midway is carried out.

3 - 28
 3.4.2

Conditioning time
TIME %A %B %C %D FLOW PRESS [LC]
0.0 30 70 0 0 1.000 5.5 2

During the conditioning time, liquid feeding is

performed under the initial conditions of the execution
step (program No. 2 in this example).

Initial
TIME %A %B %C %D FLOW PRESS [IN]
0.0 30 70 0 0 1.000 5.5 2

Liquid feeding is performed in the initial status (under

the initial conditions of program No. 2 in this example).

Gradient
TIME %A %B %C %D FLOW PRESS [GR]
0.0 40 60 0 0 1.000 5.5 2

Gradient operation (execution of program No. 2 in this

example) is performed.

Conditioning time
TIME %A %B %C %D FLOW PRESS [LC]
0.0 10 90 0 0 1.000 5.5 1

After completion of the last step of the link program,

control is returned to the first step (program No. 1 in
this example) for execution of conditioning.

Delay time (initial)

TIME %A %B %C %D FLOW PRESS [IN]
0.0 10 90 0 0 1.000 5.5 1

Liquid feeding is performed in the initial status of the

first step (program No. 1 in this example).
If the start is not input during the delay time, control
goes to column flushing ).
If the start is input during the delay time, control is
returned to ).

Column flushing
TIME %A %B %C %D FLOW PRESS [FL]
0.0 10 90 0 0 1.000 5.5 1

3 - 29
3.4 Utilities

3.4.3 Column Flush Take the following procedure for setting up automatic stop of the
Setting pump and column flushing after completion of analysis.
Where column flushing is set up, automatic washing is
performed conveniently after completion of analysis.
For instance, when a salt solvent has been used in analysis, it is
preferable to wash the column for preventing salt deposition.
Demonstrated below is an example in which column flushing is
performed with 100% solution C for a period of 30 minutes after
a lapse of 15 minutes following the end of analysis.

MONITOR
screen

UTLTY
(SETUP)

UTILITY (CONDITION = 1, UNIT = 2, FLUSH = 3

3 EVENT = 4, LINK = 5, OTHERS = 6)

Select FLUSH.

AUTO PUMP OFF (YES = 1, NO = 0)

0

Select AUTO PUMP OFF.

COLUMN FLUSH (YES = 1, NO = 0)

1

0 ENT SET DELAY TIME (1.0 - 600.0) min

600.0
Enter a period of time to be taken until the
pump is stopped after the end of analysis.
The allowable delay time range is 1.0 to
600.0.

MONITOR
screen

1 ENT

FLUSH MODE (PARAM = 1, PROGRAM NO.9 = 2)

1

Enter 1 for flushing the column under certain conditions. Enter 2

for flushing the column in gradient operation.

3 - 30
 3.4.3

1 ENT SET DELAY TIME (1.0 - 600.0) min

600.0
Enter a period of time to be taken until column
flushing is started after the end of analysis.

COLUMN FLUSH (B C D FLOW)

100 0 0 1.000
Specify column flushing conditions (mixing
percentage of solution B, mixing percentage
of solution C, mixing percentage of solution D,
flow rate). The mixing percentage of solution
A is (100 - %B - %C - %D).
In the single-liquid delivery mode, %B, %C
and %D are not indicated. In the two-liquid
high-pressure gradient operation, %C and %D
are not indicated. In the three-liquid high-
pressure gradient operation, %D is not
indicated.

FLUSHING TIME (1.0 - 600.0) min

30.0
Enter a column flushing time.
The allowable flushing time range is 1.0 to
600.0. (Example: 3 0 ENT)

MONITOR
screen

When the execution time of column flushing is

reached, column flushing is carried out according to
No.9 program.

NOTICE: Program No. 9 is used for column flushing in any

case. When column flushing is set up, program
No. 9 is automatically adopted for execution of
column flushing.
Where column flushing is specified, program
No. 9 is not available for gradient operation.
Note, however, that program No. 9 is available for
the link program function.
On termination of execution of program No. 9, the
pump is automatically stopped.

2 ENT MONITOR
screen

3 - 31
3.4 Utilities

The relationship between delay time and column flushing is as

shown below.

Delay time Column Flush

End of gradient
analysis

(1) In Case of YES for AUTO STOP

(a) Where YES is specified for COLUMN FLUSH:

Analytical Analytical Analytical Analytical

sequence 1 sequence 2 sequence 3 sequence n*

Delay time COLUMN FLUSH

Stabilization ALL END Stop of pump

time received
from L-2200

* n stands for the number of analyses specified on the autosampler

or the like.

Stabilization time represents a period of time to be taken after

the end of one analytical sequence until the start of the next
analytical sequence. After the lapse of a delay time, column
flushing is carried out.

NOTICE: If the ALL END signal is received from the L-2200

autosampler even before the end of a delay time,
column flushing is carried out.

3 - 32
 3.4.3

(b) Where NO is specified for COLUMN FLUSH:

Analytical Analytical Analytical Analytical Stop of pump

sequence 1 sequence 2 sequence 3 sequence n*

Delay time

Stabilization time
ALL END received
from L-2200

* n stands for the number of analyses specified on the

autosampler or the like.

Stabilization time represents a period of time to be

taken after the end of one analytical sequence until the
start of the next analytical sequence. After the lapse
of a delay time, the pump is forced to stop.

NOTICE: If the ALL END signal is received from the L-2200

autosampler even before the end of a delay time,
the pump is forced to stop.

(2) In Case of NO for AUTO STOP

Analytical Analytical Analytical Analytical

sequence 1 sequence 2 sequence 3 sequence n*

Pump remains in liquid feed state.

Stabilization time

* n stands for the number of analyses specified on the

autosampler or the like.

The pump remains in the liquid feed state.

NOTICE: The pump does not stop even when the ALL END
signal is received from the L-2200.

3 - 33
3.4 Utilities

3.4.4 Event Code An event can be carried out in the manual mode. When an
Setting event code is input, the event function concerned is made active.
It is thus allowed to start an external instrument, stop the pump,
and sound the buzzer.
On occurrence of a pressure limiter error, a pressure signal
indication can be given for event No. 4.

MONITOR
screen

UTLTY
(SETUP)

UTILITY (CONDITION = 1, UNIT = 2, FLUSH = 3

4 EVENT = 4, LINK = 5, OTHERS = 6)

Select any event.

EVENT SET (PRESS LIMIT = 1, EVENT CODE = 2)

1

1 ENT PRESS LIMIT (00 - 92)

6 (NOTE: EVENT NO.4)

MONITOR
screen

2 ENT SET EVENT CODE (00 - 92)

00

MONITOR
screen

3 - 34
 3.4.4

The relationship between event codes and their operations is as

follows.

Control Item Event Code Remarks

Timer 1 10 11 12 1st digit
2 20 21 22 Indicates a timer
number.
3 30 31 32 2nd digit
4 40 41 42 Indicates an
operational class
code.
Contact Start point OPEN * The contact is
open closed for one
second.
1s
Contact
Operation Signal closed
CLOSE
OFF marker*
Signal ON
Start
point

Pump 00 The pump is

stopped.
Buzzer 92 The buzzer sounds
for five seconds.

NOTICE: Where pressure limiter error setting is made, the

setting of event No. 4 in a gradient analysis
program is ignored.

3 - 35
 4.1

4. PERFORMANCE CHECK

4.1 Checking the Mixing Ratio Accuracy in a Gradient System

WARNING
Ignition of Flammable Chemicals!

Beware of ignition hazard when using flammable

chemicals such as organic solvents.
Always check the following conditions. If an abnormality
is found, stop operation immediately.
Leakage of solvent or waste solution
Leakage of solvent inside the instrument
Inadequate ventilation of the laboratory room
This instrument is not explosion-proof. Although aqueous
solvents or organic solvents having an ignition point of
70 C or higher are usable, do not use organic solvents
having an ignition point below 70 C.
When using flammable chemicals, be careful about
possible ignition due to static electricity.
Particularly when using non-conductive chemicals, employ
a conductive vessel made of metal or the like and provide
grounding connection correctly.

WARNING
Explosion of Vapor from Flammable Chemicals!

If a flammable chemical such as organic solvent leaks

from the flow path of the instrument and its vapor
concentration exceeds the explosion limit, it may cause
spontaneous combustion with dangerously explosive
results.
When using a flammable and readily volatile chemical, be
sure to check for leakage from the instrument flow path
and ventilate the laboratory room adequately.

4-1
4.1 Checking the Mixing Ratio Accuracy in a Gradient System

4.1.1 Preparation Before proceeding to performance check, be sure to remove air

Before bubbles completely from the HPLC system flow path (the solvent
Measurement filter in particular). For air purging, take the following procedure.

(1) Solvent : Methanol (guaranteed special grade)

(2) Flow rate : 2 mL/min

(3) Mixing ratio : A/B/C/D = 25/25/25/25 (%)

Under the above condition, perform air purging for at least ten
minutes (with the drain valve open).

NOTICE: 1. During low pressure gradient elution when the

mobile phases are generated by mixing by the
pump, the mobile phases may be unstable in
their ratios at each step. This instability may
be the result of air bubbles in the flow path or
contaminants adhering to the inner wall of the
flow tube. Air bubbles remain in the solvent
filters, proportioning valves or mixers in many
situations.
Flush the flow path using the following
procedures to remove the air bubbles and
contaminants and to ensure stability of the
ratios at each step.
(1) Using methanol solvent, purge the flow
path for at least ten minutes. Be sure to
remove air bubbles completely from the
flow path (the solvent filter in particular).
(2) For air purging, use the degasser.
Where a degasser is not equipped,
deaerate the solvent thoroughly to prevent
occurrence of air bubbles.
(3) Open the drain valve, and purge the
solvent flow paths A, B, C and D for 5 to
10 minutes each.
(4) Close the drain valve, and flush all the
flow paths with a mixed solvent
(containing A, B, C and D solvents at 25%
each) at a flow rate of 2 mL/min.
Flush the paths for 15 minutes or longer.

4-2
 4.1.1

(contd)

(5) Press INIT key. This allows 100% of

solvent A to flow at a rate of 1 mL/min.
Wait until the baseline stabilizes at the
zero level.
Then, start accuracy measurements of the
gradient mixing ratio.
2. When using only mobile phases A and B,
disconnect the tubes for channels C and D and
attach the furnished plugs. And when using
only mobile phases A, B and C, remove the D
channel tube and attach the furnished plug.
The air bubbles formed in the Teflon tubes that
connect the solenoid valves of unused flow
paths with the four-way joint during tubing of the
four mobile phases will flow into the flow paths
over long use, causing a variation in the mixing
ratio of solvents and possibly affecting the
retention time. So be sure to observe the
above instructions. (When unused flow paths
are connected to the four-way joint, it may be
just as effective (instead of using plugs) to
create and execute a program for also filling the
unused paths with solvent, and for supplying
solvent to the unused paths also at the end of
each analysis, in order to force out air bubbles
from the Teflon tubes connecting the solenoid
valves and four-way joint.)

C, D

Plug
810-1348

4-3
4.1 Checking the Mixing Ratio Accuracy in a Gradient System

4.1.2 Conditions (1) Measurement Conditions

Mobile phase A : Distilled water

Mobile phase B : 20 mg/L caffeine aqueous solution
Column : Connect a load resistance coil
(0.25 mm ID 10 m)
Detector : Model L-2400 UV detector at 273 nm
Time constant.............. 2 s
Output ......................... 1 AUFS/1 V
Degasser : Built-in degasser for L-2100
Data processor : Sampling period 400 ms

(2) Program Contents

LOW PRESSURE/HIGH PRESSURE GRADIENT

SYSTEM

PRESSURE MAIN : MAX MIN (MPa)

39.2 0
TIME %A %B %C %D FLOW
0.0 100 0 0 0 1.000
10.0 100 0 0 0
10.1 90 10 0 0
20.0 90 10 0 0
20.1 50 50 0 0
30.0 50 50 0 0
30.1 10 90 0 0
40.0 10 90 0 0
40.1 0 100 0 0
50.0 0 100 0 0
50.1 100 0 0 0

NOTICE: Before checking the accuracy of gradient mixing

ratio, be sure to observe the cautionary instructions
on mounting/tightening of each part.

4-4
 4.1.3

4.1.3 Mixing Ratio Calculate the mixing ratio at 10, 50 and 90% with reference to 0
Calculation to 100%.
Method
100%

90%

50%
H100
H90

H50
10%

0% H10
Mobile phase B
Start

H10
Mixing ratio: B10 = 100 (%)
H100
H50
B50 = 100 (%)
H100
H90
B90 = 100 (%)
H100

4.1.4 Specifications Low Pressure/High Pressure Gradient Elution

Within 1% when mixing ratio of mobile phase B is set to 10,

50 and 90%
(Example) B10 = 10 1% = 9 to 11%

4-5
 5.1

5. MAINTENANCE AND CHECKUP

5.1 Error Messages and Countermeasures

WARNING
Beware of Electric Shock in Contact with
Inside of Instrument!

Before removing the instrument cover for replacement or

adjustment of inside parts, be sure to turn OFF the power
switch and unplug the power cord.

NOTICE: If a failure occurs in the instrument, be sure to refer

repair servicing to qualified service personnel of
Hitachi High-Technologies Corp. Anyone other
than personnel qualified and authorized by Hitachi
High-Technologies Corp. should not attempt to
repair the instrument, for preventing a possible
hazard.

(1) On occurrence of an abnormality in the L-2100/2130 pump,

an error is indicated on the LCD (UI pad liquid crystal
display). There are two kinds of error indications, i.e.,
errors indicated according to the results of self-diagnosis
when the POWER switch is turned on (at the time of
initialization), and errors indicated due to abnormalities
encountered during operation.

5-1
5.1 Error Messages and Countermeasures

(2) Immediately after the POWER switch is turned on,

initialization is carried out in the instrument. If an
abnormality is detected during the initialization, an error
message is indicated on the LCD. Furthermore, if an
abnormality is detected during analytical operation, an error
message indication is also made on the LCD. Using the
CL key, you can clear an error message displayed on the
LCD. Note, however, that the error message appears
again unless a proper countermeasure is taken. For error
messages on units other than the L-2100, refer to the
instruction manual accompanying each unit.

Table 5-1 Error Messages in Model L-2100/2130

No. Indication on LCD Error Condition Remedy

1 CHECK The sub-pump for high- Turn on power to the sub-pump,
PERIPHERALS pressure gradient analysis is or turn it off/on again. If an error
not yet connected. occurs during high-pressure
analysis operation, turn on power
to the master pump again.
2 COMMUNICATION The sub-pump for high- Turn on power to the sub-pump,
ERROR pressure gradient analysis is turn it off/on again, or check the
not yet connected. e-line cable connection.
3 PUMP OFF BY The pump is forced to stop For restoration, press CL key.
EXTERNAL ERROR by means of the emergency
stop contact for the e-line.
4 DATA BUFFER Communication with the For restoration, press the CL key.
OVERFLOW CDS is abnormal.
5 TIME PROGRAM Communication with the For restoration, press the CL key.
BUFFER OVERFLOW CDS is abnormal.
6 PUMP A TYPE IS The pump mode setting is Turn power off, and set up the
DIFFERENT FROM wrong in high-pressure pump mode properly. Then, turn
PUMP B gradient analysis. power on again.

5-2
 5.2

5.2 Troubleshooting

Table 5-2 presents the troubleshooting procedures for the

L-2100/2130. On occurrence of a trouble, refer to this table.

Table 5-2 L-2100/2130 Troubleshooting

No. Symptom Possible Cause Remedy

1 Pressing the SET PROG The programming mode is Press the INIT key, and set
key does not set up the input not selected for the monitor up the programming mode.
mode. window.
2 Solvent does not flow. The flow rate is set at 0. Press the SET PROG or
MANUAL key, and set up a
proper flow rate.
3 Pressing the START/STOP 1) The monitor window is 1) Press the ESC key, and
key does not start gradient not selected. select the monitor
operation. window.
2) The programming mode 2) Press the INIT key, and
is not set up. set up the programming
mode.
4 Pressing the PURGE key The monitor window is not Press the ESC key, and
does not set/reset purging. selected. select the monitor window.
The instrument is waiting for
input of a numeric value.
5 After the POWER switch is The e-line cable is not 1) Connect the e-line cable.
turned on to present the connected.
initialize window, a transition 2) Turn ON the e-line setting
from the e-line connection for other units connected.
waiting window to the
monitor window does not
take place.
(In high-pressure gradient
operation)

NOTE: For troubleshooting procedures concerning other modules such as the L-2200 and L-2300,
refer to the instruction manual accompanying each module.

5-3
5.3 Periodic Maintenance and Checkup

5.3 Periodic Maintenance and Checkup

Carry out periodic maintenance and checkup properly to ensure

trouble-free reliable operation of the instrument. Table 5-3
shows a list of periodic maintenance and checkup items.

Table 5-3 Periodic Maintenance and Checkup Items

No. Item Procedure Remarks

1 Proportioning If the instrument will be left A malfunction will occur in the
valve unused for a long time, fill the flow solenoid valve unit if a solvent
path system with distilled water. crystallizes on the valve part.
2 Conventional Perform ultrasonic cleaning every If debris is accumulated in the
mixer four months. mixer, air bubbles may be
produced.

5-4
 5.4.1

5.4 Gradient Mixer

In the gradient system, the gradient mixer is used for mixing two
or more solvents. For low-pressure or high-pressure gradient
analysis, select the conventional mixer or semi-micro mixer
according to the flow rate to be used.

5.4.1 Conventional For analysis at a flow rate of 1.8 mL/min or higher, it is

Mixer recommended to use the large-type mixer that is available
separately.
For details of usage of the mixers, refer to 3.2.2.

(1) Structure of Conventional Mixer

The conventional mixer is provided with a chamber

containing glass beads.

(2) Maintenance and Checkup

(a) Using a wrench, remove the filter from the joint.

(b) Remove glass beads from the joint, and put them into
a beaker (50 to 100 mL). Then, pour a detergent
(such as a mobile phase) into the beaker.

(c) Place the filter and joint into another beaker, and pour
a detergent into it.

(d) Set the two beakers in an ultrasonic cleaner, and

perform cleaning for five to ten minutes.

(e) Assemble the conventional mixer in the reverse order

of removal.

NOTICE: When disassembling the conventional mixer, take

care not to drop the glass beads.

5-5
 6.1

6. SPARE PARTS

Described below are the spare parts necessary for ensuring

optimum use of this system (part numbers, names, configuration,
specifications). The standard useful life of each spare part is
based on the condition that methanol is fed at a flow rate of
1 mL/min with a pressure of 19.6 MPa. The recommended
frequency of replacement of each spare part is based on the
condition that the instrument is operated 240 days per year, for
eight hours per day. The standard useful life and
recommended frequency of replacement of each spare part
varies significantly with the analytical conditions such as solvent,
flow rate and pressure. So, check them in consideration of
actual analytical conditions.
Use the following list when ordering spare parts.

6.1 Gradient Spare Parts for L-2100

Component Standard
Configuration/
Part No. Part Name (Recommended frequency Useful
Specifications
of replacement) Life
810-1031 Flow path filter Outlet of mixer 1920
(One year) hours

885-1505 Packing Four-way joint 1920

(One year) hours

890-6502 Solenoid valve Proportioning valve 1920

(For A, B, C and D) hours
This part is a consumable.
If liquid leakage is found on
the threaded part, replace
the part with a new one.

6-1
 7.1

7. SPECIFICATIONS

7.1 Model L-2100 Low-Pressure Gradient System

(1) Number of eluents mixable

Up to four liquids

(2) Principle of mixing

Valve opening/closing time control

(3) Mixing ratio range

0 to 100% (in increments of 1%)

(4) Flow rate range

0.001 to 2.499 mL/minute, in increments of 0.001

mL/minute

(5) Recommended flow rate range

In use with conventional mixer (capacity: 1 mL)

0.4 to 1.8 mL/minute
In use with semi-micro mixer (capacity: 0.2 mL)
0.4 mL/minute or less

(6) Mixing ratio accuracy

1% (in a flow rate ranging from 0.05 to 1.5 mL/minute)

(7) Gradient profile

Linear, multi-stepped

(8) Number of steps

Up to 92 steps per program

Up to 100 steps per 9 programs

(9) Programmed time

Up to 600.0 minutes (in increments of 0.1 minute)

7-1
7.1 Model L-2100 Low-Pressure Gradient System

(10) Program input items

(a) Time

(b) B-C-D mixing ratio

(c) Flow rate

(d) Timer (marker) output

(e) Stop of pump (buzzer)

(11) Number of programs

Up to 9 programs

(12) Display (option)

LCD display (80 characters across by two lines)

(13) Displayed items (option)

(a) Input parameters

(b) Monitored data (elapsed time, mixing ratio, flow rate,

pressure, timer, etc.)

7-2
 7.2

7.2 Model L-2100 High-Pressure Gradient System

(1) Number of eluents mixable

Up to two liquids

(2) Principle of mixing

Two-pump flow rate control

(3) Mixing ratio range

0 to 100% (in increments of 0.1%)

(4) Flow rate range

0.001 to 2.499 mL/minute, in increments of

0.001 mL/minute

(5) Recommended flow rate range

0.05 to 2.0 mL/minute

(6) Mixing ratio accuracy

1% (in a flow rate ranging from 0.05 to 2.0 mL/minute)

(7) Gradient profile

Linear, multi-stepped

(8) Number of steps

Up to 92 steps per program

Up to 100 steps per 9 programs

(9) Programmed time

Up to 600.0 minutes (in increments of 0.1 minute)

7-3
7.2 Model L-2100 High-Pressure Gradient System

(10) Program input items

(a) Time

(b) B-C-D mixing ratio

(c) Flow rate

(d) Timer (marker) output

(e) Stop of pump (buzzer)

(11) Number of programs

Up to 9 programs

(12) Display (option)

LCD display (80 characters across by two lines)

(13) Displayed items (option)

(a) Input parameters

(b) Monitored data (elapsed time, mixing ratio, flow rate,

pump-A pressure, pump-A event output)

7-4
 7.3

7.3 Model L-2130 Low-Pressure Gradient System

(1) Number of eluents mixable

Up to four liquids

(2) Principle of mixing

Solenoid valve opening/closing time control

(3) Mixing ratio

0 to 100% (in UI pad setting, indicated in increments of
1%)
Accuracy in mixing ratio
Within 1% (in use with degasser, at 10%, 50%, 90% for
water - caffeine aqueous solution)

(4) Flow rate range

0.001 to 9.999 mL/minute, in increments of 0.001

mL/minute

(5) Recommended flow rate range

In use with conventional mixer (capacity: 1 mL)

0.4 to 1.8 mL/minute
In use with semi-micro mixer (capacity: 0.2 mL)
0.4 mL/minute or less

(6) Gradient profile

Linear, multi-stepped

(7) Number of programs

9 programs

(8) Number of steps

Up to 92 steps per program

(9) Up to 100 steps per 9 programs

7-5
7.3 Model L-2130 Low-Pressure Gradient System

(10) Programmed time

Up to 600.0 minutes (in increments of 0.1 minute)

(11) Program input items

(a) Time

(b) B-C-D mixing ratio

(c) Flow rate

(d) Event timer output

(e) Stop of liquid feed

(f) Buzzer

(12) Displayed items with UI pad (option)

(a) Input parameters

(b) Monitored data (elapsed time, mixing ratio, flow rate,

pressure, event output)

7-6
 7.4

7.4 Model L-2130 High-Pressure Gradient System

(1) Number of eluents mixable

Up to two liquids

(2) Principle of mixing

Two-pump flow rate control

(3) Mixing ratio

0.0 to 100.0% (in increments of 0.1%)
Accuracy in mixing ratio
Within 1% (in use with degasser, at 10%, 50%, 90% for
water - 0.1% acetone aqueous solution)

(4) Flow rate range

0.001 to 9.999 mL/minute, in increments of 0.001

mL/minute

(5) Recommended flow rate range

0.2 to 8.0 mL/minute

(6) Gradient profile

Linear, multi-stepped

(7) Number of programs

9 programs

(8) Number of steps

Up to 92 steps per program

(9) Up to 100 steps per 9 programs

(10) Programmed time

Up to 600.0 minutes (in increments of 0.1 minute)

7-7
7.4 Model L-2130 High-Pressure Gradient System

(11) Program input items

(a) Time

(b) B mixing ratio

(c) Flow rate

(d) Event timer output

(e) Stop of liquid feed

(f) Buzzer

(12) Displayed items with UI pad (option)

(a) Input parameters

(b) Monitored data (elapsed time, mixing ratio, flow rate,

pump-A pressure, pump-A event output)

7-8