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228-92687

Sep. 2017

SHIMADZU HIGH PERFORMANCE LIQUID CHROMATOGRAPH

Prominence-i
LC-2030 Plus
LC-2030C Plus
LC-2030C 3D Plus

Integrity Guide
Read this manual thoroughly before you use the product.
Keep this manual for future reference.
This page is intentionally left blank.
Introduction
Read this Instruction Manual thoroughly before using
the product.
Thank you for purchasing this product.
This manual describes the information about "Testing" for verifying that this product
meets "CHECK CRITERIA" and about "calibration" for adjustment so that this product
meets the CHECK CRITERIA, in order to secure the reliability of analysis data. Read this
manual thoroughly before using the product and operate the product in accordance with
the instructions in this manual. The following instruction manuals are included with the
product. PDF documents are included in the instruction manual CD-ROM.

System Guide Describes the basic operation and troubleshooting for this
(Booklet/PDF) product.
This instruction manual. Describes "validation" for verifying that
Integrity Guide this product meets "CHECK CRITERIA" and "calibration" for
(PDF) adjustment so that this product meets the CHECK CRITERIA, in
order to secure the reliability of analysis data.
Describes the operation of the operations screen and the
Operation Guide
operational method for the Web screen, iPhone, and iPad for
(PDF)
this product.
Maintenance Guide Describes the maintenance for this product, including
(PDF) inspection, part replacement, and rinsing.

Read "Introduction" in this section thoroughly before using the product. "Introduction"
describes the information about the warranty, after-sales service, safety instructions and
precautions to ensure safe operation of the instrument.
For information on the operation, hardware validation, and accessories and options for
this product, refer to "System Guide", "Operation Guide" and "Maintenance Guide".
Keep this manual for future reference.

Important
• If the user or usage location changes, ensure that this manual is always kept
together with the product.
• If this manual or a product warning label is lost or damaged, immediately contact
your Shimadzu representative to request a replacement.
• To ensure safe operation, read "Safety Instructions" P.iv thoroughly before using
the product.
• To ensure safe operation, contact your Shimadzu representative if product
installation, adjustment, re-installation (after the product is moved), or repair is
required.

ã 2017 Shimadzu Corporation. All rights reserved.


Original version is approved in English.

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Introduction

Notice
• Information in this manual is subject to change without notice and does not represent
a commitment on the part of the vendor.
• Any errors or omissions which may have occurred in this manual despite the utmost
care taken in its production will be corrected as soon as possible, although not
necessarily immediately after detection.
• All rights are reserved, including those to reproduce this manual or parts thereof in
any form without permission in writing from Shimadzu Corporation.
• Some portions of illustrations in this manual might be different from the instrument
used.
• Microsoft and Windows are registered trademarks of Microsoft Corporation in the
United States and/or other countries.
iPhone and iPad are registered trademarks of Apple Inc. in the United States and/or
other countries.
Other company names and product names mentioned in this manual are trademarks or
registered trademarks of their respective companies. The ä and â symbols are omitted
in this manual.
• Prominence-i is registered trademarks of Shimadzu Corporation in the United States and
Europe (Office for Harmonization in the Internal Market).

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Indications Used in This Manual

Indications Used in This Manual


Warnings, cautions, and notes are indicated using the following conventions:

Indication Meaning

Indicates a potentially hazardous situation which, if not


avoided, could result in serious injury or possibly death.

Indicates a potentially hazardous situation which, if not


avoided, may result in minor to moderate injury or equipment
damage.
Emphasizes additional information that is provided to ensure
the proper use of this product.

The following symbols are used in this manual:

Indication Meaning

Indicates an action that must not be performed.

Indicates an action that must be performed.

Indicates information provided to improve product performance.

Indicates the location of related reference information.

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Introduction

Safety Instructions
To ensure safe product operation, read these important safety instructions carefully
before use and follow all WARNING and CAUTION instructions given in this section.

n Product Applications

Use this instrument ONLY for the intended purpose.


This instrument is a high performance liquid chromatograph system. Using this
instrument for any other purpose could cause accidents.

Safety regulations and standards


For notifications on installation and safety controls, follow the necessary
procedures in compliance with the laws and regulations applicable in the
country where the product is used.

n Installation Site

Use of open flame in the vicinity of this instrument


must be strictly prohibited.
Do NOT install the instrument in the same room
with any other equipment that emits. Otherwise
could potentially emit sparks, since sparks could
cause a fire.
High performance liquid chromatograph uses large
amounts of flammable organic solvents.
Provide fire extinguishers for use in case of fire.

The room where the instrument is installed should


be well ventilated.
Otherwise, solvent vapors could cause poisoning or
ignite and cause a fire. The solvents used in high
performance liquid chromatograph are flammable
and toxic.

Provide sink washing equipment as close to the


instrument as possible.
If solvent gets into the eyes or onto the skin, it must
be flushed away immediately.

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Safety Instructions

Avoid installation sites that are exposed to corrosive gases or excessive dust.
These adverse conditions may be detrimental in maintaining the instrument's
performance and may shorten its service life.

Keep away from equipment generating strong magnetic fields.


Do NOT install the instrument near equipment that generates strong magnetic
fields. If the power supply line is subject to high electrical noise, install a surge
protector.

When transporting the instrument, do not hold it by the front panel.


Otherwise, it may cause the front panel to get damaged or detached.

During installation, consider the entire mass of this system combined with
other components.
• Model without a sample cooler: 58 kg
• Model with a sample cooler: 63 kg
The mass of this system is as described above. Please set up this instrument in
consideration of the entire mass combined with other components, PC, the
monitor, and the printer, etc.
"7 Technical Information" in the System Guide

The lab table on which this instrument is installed


should be strong enough to support the total mass
of the system. It should be leveled, stable and have
depth of at least 600 mm.
Otherwise, the instrument could tip over or fall off
the table.

Keep at least 100 mm between the rear of the


instrument and the wall.
This allows for sufficient air circulation and
ventilation from the grille to provide cooling and
prevent the instrument from overheating and
impairing the performance.

Install the instrument in a location that satisfies the following conditions to


preserve its performance:
• The room temperature is maintained between 4 and 35 °C, with minimal
temperature variation during a day.
• Air currents from heating or air conditioning equipment are not directed
onto the instrument.
• Ensure that Instrument is not exposed to direct sunlight.
• There is no vibration.
• Humidity is maintained within 20 to 85 %.
• There is no condensation.
• The location conforming to the installation environment (IEC)
(Installation category II, Pollution degree: 2, Altitude: Up to 2000 m, Indoor)

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Introduction

Install the product in locations where the power switch can be easily operated.
The power switch must be turned off in case of an emergency.

n Installation

To ensure safe operation, contact your Shimadzu representative if product installation,


adjustment, or re-installation (after the product is moved) is required.

Do NOT place heavy objects on the power cord. Do


NOT keep any hot items in the vicinity.
Do NOT modify the cord in any way. Do NOT bend
it excessively.
Otherwise, fire, electric shock or malfunction may
occur.
If the cord is damaged, contact your Shimadzu
representative.

Use only the specified cable.


Arrange the cables as specified.
Otherwise, fire, electric shock or malfunction may
occur.

Do NOT insert the adapter's grounding lead into


the power outlet or allow it to make contact with
the power outlet.
Otherwise, fire or electric shock may occur.

Do NOT connect the grounding lead to the gas


tube, the water service tube or the telephone line.
Otherwise, accidents or malfunction may occur.

To unplug the instrument, pull the plug itself, NOT


the cord.
Before connecting the cable, turn off the power and
unplug the instrument.
Otherwise, fire, electric shock or malfunction may
occur. If the cord is damaged, contact your
Shimadzu representative.

Take measures to prevent the instrument from falling in the event of an


earthquake or other disaster.
Strong vibrations could cause the instrument to fall over, resulting in injury.

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Safety Instructions

Ground the instrument.


Grounding is necessary to prevent electric shock in the event of an accident or
electrical discharge, and important for ensuring stable operation.
To prevent electric shock and to maintain stability in operation of the product,
be sure to ground the product.
The product will be grounded when the provided 3-wired power plug is
inserted into a 3-wired power socket equipped with a ground terminal.

Connect the instrument only to a power supply of indicated voltage in


consideration of total power consumption of this instrument, other LC
components, PC, monitor, and printer, etc.
Otherwise, fire or electric shock could result.
Check that the power supply voltage is stable and that its current capacity is
sufficient to operate all the components of the system. If not, the instrument
will not operate at its rated performance, affecting not only this instrument,
but other instruments connected to the same power supply. Connect the
instrument to a power supply that complies with the capacity and use a power
cord that complies with the capacity.
Please refer to the following for power supply voltage and power consumption
of this product.
"7 Technical Information" in the System Guide

When installing the instrument, be careful not to


pinch your fingers between the system components.
Otherwise, injury may result from pinching your
fingers, because the clearance between the
components is only 20 mm.

When opening the doors, be careful not to pinch


your fingers as this could result in injury.

Before plugging in the instrument, make sure that


the main power switch on the side of the
instrument is OFF.

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Introduction

n Operation

Do NOT use flammable sprays (hair sprays,


insecticide sprays, etc.) near the instrument.
Otherwise, they could ignite and cause a fire.

Never use a cracked reservoir bottle.


If a helium degassing unit is used, pressure is
exerted on the reservoir bottles and may cause
cracks in the bottles. It could break the reservoir
bottles and cause injury.

Do NOT apply water to office equipment such as the PC as well as the


instrument.

Before removing any high pressure flow line part, such as tubing, the sample
loop, the needle, and the needle seal, follow the safety precautions below:Be
sure to stop the pump and confirm the pressure of the mobile phase has fallen
to zero.
Removing any of these parts while high pressure remains could cause the
mobile phase and the clogging particles to eject rapidly and injury could occur.
Confirm that the pressure has fallen to zero before removing these parts.
If the pressure has not fallen to zero, the high-pressure flow line might be
clogged. After confirming the end of the drain tubing is in the waste container,
open the drain valve and release the pressure of the mobile phase.

Take thorough measures to prevent buildup of


static electricity.
Static electricity could result in fires or explosion.
"Static Electricity Precautions" P.xi

Always wear protective gloves and protective


goggles when handling solvents and samples.
If solvent gets into the eyes, it can result in
blindness.
Should solvent get into the eyes, flush immediately
with large amounts of water and get medical
attention.

Do NOT bend any tubing of the instrument at the same position repeatedly.
Otherwise, a rupture or cracks of the tubing are likely to occur, and might
result in solvent leaks.

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Safety Instructions

n Inspection and Maintenance

Never remove the main cover.


Otherwise, this may cause injury or malfunctioning of the instrument.
The main cover does not need to be removed for routine maintenance,
inspection and adjustment. Have your Shimadzu representative perform any
repairs requiring removal of the main cover.

Do NOT allow spilled water to remain on the instrument surface, and do not
use alcohol or thinner-type solvents to clean the surfaces.
Otherwise, these can cause rusting and discoloration.

If the power cord plug gets dusty, remove the plug from the power outlet
and wipe away the dust with a dry cloth.
Otherwise, fire may occur.

Replacement parts must be of the specifications given in "Component Parts"


or "Maintenance Parts" of the instruction manuals such as the System Guide
and the Maintenance Guide.
Use of any other parts may result in instrument damage and malfunction.

Dispose of the waste liquid properly and in accordance with the instruction
by your administrative department.

n Repair, Disassembly and Modification

Do NOT modify or disassemble the product without permission.


Otherwise, electric shock or short-circuit accidents may occur. This may also
cause injury or malfunctioning of the instrument.

When repair is necessary, request through your Shimadzu representative.


Failing to do so may result in an ignition, electric shock, or injury.

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Introduction

n In an Emergency

If any problem is detected, such as a burning smell, take the following action:

Emergency Shutdown Procedure

1 Turn the power to the instrument OFF.

2 Disconnect the power cable at the rear of the instrument.

When the instrument is used again, inspect the instrument and, if necessary, contact your
Shimadzu representative to request servicing.

n During a Power Outage

Take the following measures in the event of a power outage.

1 Turn the power to the instrument OFF.

2 After confirming all related items in this section "Installation" P.vi and "Operation"
P.viii, use the standard startup procedure to start the instrument.

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Static Electricity Precautions

Static Electricity Precautions


A liquid chromatograph (LC) uses flammable organic solvent(s) as the mobile phase. LC
systems are also often used where large amount of flammable substances are present.
Thus, an accident can produce large scale damage. Operators must be constantly on guard
against accidents involving fire or explosion.
The major cause of these accidents is static electricity. Devising preventative measures for
static electricity can be difficult, because the symptoms before an accident vary and can
be hard to detect, since such accidents occur as a result of several simultaneous incidents.
Recommended methods for preventing static electricity accidents are provided below.
Take thorough safety measures based on this information.

n Typical Cause of Static Electricity Accidents

Static electricity accidents are generally caused by this sequence of events:

Generation of static When liquid is passed through thin tubing at high flow
electricity rates, as in liquid chromatograph, the electrostatic charges
of the flowing matter generate static electricity.

Accumulation of static If electrostatically charged liquid is allowed to accumulate


electricity in an electrically insulated container, the charge will
gradually increase, and can eventually reach several
È thousand volts.
Release of energy through If this happens and an electrical conductor is brought
electrical discharge within a certain distance of the container, an electrical
discharge will occur, releasing thermal energy which will
È
ignite any flammable gas of sufficient density in the
Ignition of flammable vicinity.
substances

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Introduction

Preventing Static Electricity Accidents

The best way to prevent static electricity accidents is simply to prevent the occurrence
and accumulation of electrostatic charges.

It is important to take multiple preventive measures simultaneously.


If large amounts of flammable solvents are collected in a large container,
implement preventative measures 1, 2, and 3 below.

Keep the room at a proper humidity.


Ambient humidity exceeding 65 % will prevent static.

n Preventive Measure 1

Use a metal container for the waste liquid, and ground the container.
This will ensure that the electrical charges of the container and liquid pass to the ground.

Accessories for this measure

(1) Grounding wire with clip Part No. S228-21353-91


(2) 18-L metal container Part No. S038-00044
(3) 4-L metal container Part No. S038-00043-01

Be sure to ground the metal waste container properly.


If the grounding wire is not properly attached or connected to the ground,
static electricity can build up in the container.

Be sure to verify, using a tester, that the waste container is properly grounded.
Some metal containers have surfaces that are oxidized or laminated. Such a
container may not conduct electricity.

If the liquid to be drained into the waste container is virtually nonconductive


(10-10 S/m or less), it will be necessary to add properly conductive, and
therefore safe, liquid to the tank.
This conductive liquid may be added to the waste container beforehand.

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Static Electricity Precautions

n Preventive Measure 2

Cover the spaces between the tubing and the sides of the inlet and outlet openings of
the waste container with caps or other protective covering. This will prevent any sparks
generated outside the container from getting inside.

Accessories for this measure


Caps for 18-L or 4-L container (with three 3-mm diameter openings)
Part No. S228-21354-91

n Preventive Measure 3

Keep electrostatically charged objects, including the human body, away from the waste
liquid container.
To prevent electrostatic charging of the human body, take the following precautions:
• Wear anti-static clothing and shoes.
• Ground the human body with anti-static wrist straps. (For safety, the wrist strap should
be connected to the ground using an intervening resistor of about 1 MW.)

• Spread anti-static matting or the like on the floor, to make the floor conductive.

Persons who have not taken anti-static precautions should touch some
grounded metal object before coming near the waste liquid container, in order
to drain static charges.

n Preventive Measure 4

Use tubing with an inner diameter of at least 2 mm for drain lines with high flow rates.

Periodically check the tubing connections for leaks.


Air bubbles in liquid can multiply the electrostatic charge by a factor of 20,
30 or more.

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Introduction

n Preventive Measure 5

If it is not possible to use a conductive waste liquid container, take the following
precautions:
Ensure that the end of the inflow tubing is always submerged inside the container. Also,
place some type of grounded metal object, such as a ground wire connected to the
instrument, into the liquid.

For low conductivity (less than 10-10 S/m) liquids, take preventive measures 1
to 4.
For low conductivity (less than 10-10 S/m) liquids, preventive measure 5 has no
effect.
Use as small a container as possible to minimize damage in the event of fire.

For Reference
Anti-static equipment (anti-static clothing, shoes and matting) and charge measurement
equipment (potentiometer) are sold by specialty manufacturers.

xiv Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus


Precautions for Mobile Phase Selection and Use

Precautions for Mobile Phase Selection and Use

Do NOT use resin parts for the high-pressure tubing while pumping at high
pressures.
Pumping at high pressure may cause resin tubing to be ruptured or
disconnected, which could result in mobile phase leaks.
Please note the maximum withstand pressure of each part when resin parts
are used for the high-pressure tubing.

Do NOT use highly volatile acids, such as acetic acids in high concentration
(10 % to 50 %) or 1 % TFA (trifluoroacetic acid) solution, as a mobile phase
or rinse solution of the autosampler continually.
Doing so could cause metallic parts in the instrument to corrode. If such liquids
have been used for analysis, purge the mobile phase or rinse solution from the
flow line with distilled water or other liquid that is less corrosive. Also, turn
the autosampler off after analysis and open the front door slightly to let the
vapor release from the inside of the instrument.

Do not use solutions of pH 13 or more.


Some types of mobile phases may damage the flow cell quartz if used for a
long period of time at pH10 or more, resulting in transformation of the
transmission characteristics. After using this type of mobile phase, pump HPLC
grade pure water or other liquid to rinse the flow cell.

The following solvent could damage the system. Never use these solvents in
the degassing unit.

• HFIP (Hexafluoroisopropanol) • Perfluoro 1-methyldecalin


• HF (Hydrogen fluoride) • Perfluoro dimethyldecalin

• Freon 113 • Perfluoro methyl-cyclohexane

• Fluorinert FC-40 • Perfluoro dimethyl-cyclohexane

• Fluorinert FC-72 • AK-225

• Fluorinert FC-75 • Nitric acid with a concentration of 30 % or


more
• Perfluoro benzene
• Sulfuric acid with a concentration of 40 %
• Perfluoro octane or more
• Perfluoro decalin • Hydrogen peroxide

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Introduction

If PEEK resin parts are used for tubing connections, do not use the following
mobile phases.
These mobile phases weaken PEEK resin, which may lead to cracked tubing and
mobile phase leaks.

• Concentrated sulfuric acid • Dichloromethane


• Concentrated nitric acid • Chloroform
• Dichloroacetic acid • Dimethyl sulfoxide (DMSO)
• Acetone*1 • Fluorine organic solvents such as
• Tetrahydrofuran (THF) hexafluoroisopropanol (HFIP)
*1 There is no problem with temporarily using low-concentration aqueous solutions
with an acetone concentration of 0.5 % or less, e.g. for the purpose of checking
the performance of the gradient.

• Use only HPLC grade or comparable mobile phase, and filter it with a filter
of 0.45 µm mesh or finer before use to remove particulates and foreign matter.
• Halogen ions can corrode the stainless steel material (SUS316L) used in the
plumbing, so if such materials are used for the wetted parts of the equipment,
avoid as much as possible mobile phases that contain halogen ions, including
KCl, NaCl and NH4Cl or mobile phases that generate halogen ions in certain
reactions. If such mobile phases must be used, clean all flow lines thoroughly
with distilled water immediately after analysis.
• Avoid ammonia aqueous solution at a concentration exceeding 0.1 %. It may
damage the plunger of the pump and shorten the service life of the plunger
and the plunger seal.
• When an absorbance detector or photodiode array detector is used for
high-sensitivity analysis, be sure to use HPLC grade mobile phases that have a
low absorptivity of UV rays.
• Always degas the mobile phase, as air bubbles may tend to form during solvent
mixing or during temperature or pressure changes. Air bubbles may cause
pump malfunctions and detector signal noise.
• Understand the properties, including boiling points, firing point and viscosities,
of the mobile phase.

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Warning Labels

Warning Labels
In order to ensure safety, warning labels are attached in places requiring caution.
If a warning label is lost or damaged, obtain a new label through your Shimadzu
representative and attach it in the correct position.

Warning
No. Description
Label
(Part No. S037-72999-02)
1 During operations such as calibration, the autosampler operates even
with the front panel open. Do NOT touch the needle moving parts.

(Part No. S228-57699)


2 Do NOT forget to tighten tubing completely. Otherwise, leakage
may occur.

(Part No. S206-77586)


3 Be careful not to touch when the operating temperature of the
column oven is high (60 °C or over).

(Part No. S037-70854-02)


4 The lamp emits ultraviolet light when illuminated. Do NOT look
directly at the lamp light.
(Part No. S037-72999-12)
Allow the lamp and lamp mounting parts to cool down adequately
5
before replacing the lamp. The lamp compartment is extremely hot
just after turning off the lamp.

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Introduction

Warning
No. Description
Label
(Part No. S228-53303)
If a drain tube is not connected, the rinse solution may leak,
6 resulting in malfunction of the instrument. When a model with a
sample cooler is used, the condensation water may leak, resulting
in malfunction of the instrument.
(Part No. S228-53303)
7 If a drain tube is not connected, leaked solutions will not drain,
resulting in malfunction of the instrument.
(Part No. S228-53303)
If the column oven temperature is set to below the room
8 temperature, condensation may occur. If a drain tube is not
connected, leakage may occur, resulting in malfunction of the
instrument.
(Part No. S228-53303)
9 Be careful of dirt on the air filter. If the filter is clogged with dust,
the instrument's performance may be impaired.

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Warranty

Warranty
Shimadzu provides the following warranty for this product.

1. Period:
Please contact your Shimadzu representative for information about the period of this warranty.

2. Description:
If a product/part failure occurs for reasons attributable to Shimadzu during the warranty period,
Shimadzu will repair or replace the product/part free of charge. However, in the case of products
which are usually available on the market only for a short time, such as personal computers and
their peripherals/parts, Shimadzu may not be able to provide identical replacement products.

3. Limitation of Liability:
(1) In no event will Shimadzu be liable for any lost revenue, profit or data, or for special, indirect,
consequential, incidental or punitive damages, however caused regardless of the theory of
liability, arising out of or related to the use of or inability to use the product, even if Shimadzu
has been advised of the possibility of such damage.

(2) In no event will Shimadzu's liability to you, whether in contract, tort (including negligence), or
otherwise, exceed the amount you paid for the product.

4. Exceptions:
Failures caused by the following are excluded from the warranty, even if they occur during the
warranty period.

(1) Improper product handling

(2) Repairs or modifications performed by parties other than Shimadzu or Shimadzu designated
companies

(3) Product use in combination with hardware or software other than that designated by Shimadzu

(4) Computer viruses leading to device failures and damage to data and software, including the
product's basic software

(5) Power failures, including power outages and sudden voltage drops, leading to device failures
and damage to data and software, including the product's basic software

(6) Turning OFF the product without following the proper shutdown procedure leading to device
failures and damage to data and software, including the product's basic software

(7) Reasons unrelated to the product itself

(8) Product use in harsh environments, such as those subject to high temperatures or humidity levels,
corrosive gases, or strong vibrations

(9) Fires, earthquakes, or any other act of nature, contamination by radioactive or hazardous
substances, or any other force majeure event, including wars, riots, and crimes

(10) Product movement or transportation after installation

(11) Consumable items


Recording media such as CD-ROMs are considered consumable items.

* If there is a document such as a warranty provided with the product, or there is a separate contract
agreed upon that includes warranty conditions, the provisions of those documents shall apply.

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Introduction

After-Sales Service and Availability of Replacement Parts


n After-Sales Service

If any problem occurs with this product, perform an inspection and take appropriate
corrective action as described in "6 Troubleshooting" of the System Guide.
If the problem persists, or the symptoms are not covered in the troubleshooting section,
contact your Shimadzu representative.

n Replacement Parts Availability

Replacement parts for this product will be available for a period of seven (7) years after
the product is discontinued. Thereafter, such parts may cease to be available.
Note, however, that the availability of parts not manufactured by Shimadzu shall be
determined by the relevant manufacturers.

n Hardware Validation

Each unit in the instrument and the entire LC system should be checked periodically to
ensure that they function normally, else the analysis data may not be reliable.
Therefore, it is necessary to carry out periodic hardware validation and keep records of
the validation.
Before shipment from the factory, this instrument was rigorously inspected. The results
are summarized in the Inspection Certificate accompanying the instrument. To inspect the
instrument performance after installation, repeat the Hardware Validation as described
in "2 Validation" P.2.

n Hardware Validation Contract

This is a contract under which a qualified Shimadzu-approved engineer performs periodic


validation, and provides reports of the results.
Details of the contract can be obtained from your Shimadzu representative.

Maintenance, Inspections, and Adjustment


In order to maintain the instrument's performance and obtain accurate measurement
data, daily inspection and periodic inspection/calibration are necessary.
• For daily maintenance, inspection, and replacement parts, refer to the System Guide
and Maintenance Guide.
• Periodic inspection/calibration should be requested to your Shimadzu representative.
• Replacement cycles described for periodic replacement parts are rough estimate.
Replacement may be required earlier than the described replacement cycles depending
on usage environment and frequency.

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Disposal Precautions

Disposal Precautions
When disposing of the instrument, contact your Shimadzu representative.
If you dispose them yourself, do so in accordance with the processing standards
determined by law, separately from general industrial waste and household garbage.

Precautions on Handling Deuterium (D2) Lamp

n When Disposing of the Lamp

If the deuterium (D2) lamp should be broken or its life is finished, dispose of the lamp
separately from general garbage. When disposing of the deuterium (D2) lamp provided
from Shimadzu Corporation, select a method, which will not harm the environment or
cause bodily injury. Consult your local Government Agencies for a proper disposal method.

The materials of deuterium (D2) lamp are as follows:


• Metals (Tungsten, Aluminum)
• Quartz glass
• Ceramic
• Plastic

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Contents
Introduction Indications Used in This Manual ......................... iii
Safety Instructions ............................................. iv
Static Electricity Precautions ............................... xi
Precautions for Mobile Phase Selection and Use xv
Warning Labels .............................................. xvii
Warranty ........................................................ xix
After-Sales Service and Availability of
Replacement Parts ............................................ xx
Maintenance, Inspections, and Adjustment ......... xx
Disposal Precautions ........................................ xxi

1 Overview

2 Validation 2.1 Validation Overview ............................................ 2


2.2 System Check ...................................................... 5
2.2.1 Processing the System Check Using the Instrument ........... 5

2.2.2 Processing the System Check Using PC, iPad, or iPhone ..... 8
2.2.3 List of System Check Results ................................. 12

2.2.4 Saving the Results of a System Check ....................... 21

2.3 Automatic Validation ......................................... 23


2.3.1 Processing Automatic Validation .............................. 23
2.4 Performance Check ............................................ 30
2.4.1 Displaying the Items for Performance Check ................ 30
2.4.2 Pump .......................................................... 32
2.4.3 Autosampler ................................................... 44
2.4.4 Column Oven ................................................. 50
2.4.5 UV Detector ................................................... 56
2.4.6 PDA Detector .................................................. 68

2.4.7 Others ......................................................... 78


2.5 Other Validations .............................................. 83
2.5.1 Analysis Reproducibility Test .................................. 83
2.5.2 Carryover Test ................................................. 92

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Contents

2.5.3 UV Detector Absorbance Linearity Test ...................... 94


2.5.4 PDA Detector Absorbance Linearity Test ..................... 97
2.6 Check the Hardware Validation Results ............. 100

3 Calibration 3.1 Calibration Overview ........................................ 102


3.2 Hardware Calibration ....................................... 102
3.2.1 Displaying the Hardware Calibration Items ................. 103

3.2.2 Pump ........................................................ 105


3.2.3 Autosampler ................................................. 112
3.2.4 Column Oven ................................................ 121

3.2.5 UV Detector ................................................. 129


3.2.6 PDA Detector ................................................ 136

3.2.7 Leak Sensor Calibration ..................................... 146


3.3 System Adjustment .......................................... 152
3.3.1 Displaying the System Adjustment Items ................... 152
3.3.2 Adjusting the Rack Positions ................................ 155

3.3.3 Adjusting the Injection Port Position ....................... 163

Index ......................................................... 167

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1 Overview
In order to assure the reliability of analysis data, "Validation" checks that the CHECK
CRITERIA set in each unit are met. 1
Because the performance of an analyzer will vary over time with the deterioration of
consumable parts, the test should be conducted periodically from the time the instrument
is introduced until the instrument is disposed of.
When the CHECK CRITERIA are not met, perform replacement of consumable parts, failure
diagnoses, or calibration.
Calibration should be performed in order to adjust the instrument to meet the CHECK
CRITERIA.
When the CHECK CRITERIA are not met even if the above corrective actions are
performed or when the cause is unknown, contact your Shimadzu representative.

n Test and calibration flow

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2 Validation
2.1 Validation Overview

n Types of validation

Validations using the instrument include performance checks and automatic validations
processed from the instrument's Operations screen and validations processed using
external processing instruments. System check allows the user to list and check validation
results or the operating conditions of consumable parts.

The user can also check the histories of automatic validations and performance checks via
the [Maintenance] screen in the [Log] tab. The user can also check the operating condition
of a consumable part on the [Maintenance] screen.

Operation Guide "2.2 Maintenance Screen"

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2.1 Validation Overview

Item Explanation Implementation Ref.


Displays the results of performance checks
or automatic validation or the operating
System Before processing an
conditions of consumable parts in a list. P.5
Check analysis.
The list is automatically updated by the
instrument.
Fully automates the testing of items that
During periodic
do not require manual operation in the
Automatic inspection.
performance check. P.23
Validation When an abnormality is
This is processed by operating the screen
found in the instrument. 2
of the instrument.
During periodic
Processed for each item.
Performance inspection.
This is processed while operating the P.30
Check When an abnormality is
screen of the instrument.
found in the instrument.
During periodic
Other inspection.
Processed for each item. P.83
Validations When an abnormality is
found in the instrument.

The user should document and record the results of conducted checks for later
reference. The user can save the automatic validation and performance check
results from the PC Web screen and print them from the application software.

"2.2.4 Saving the Results of a System Check" P.21

n Validation items

Validation items include "performance checks" and "automatic validations" with


procedures displayed on the screen, and "other validations" that have no items on the
screen.
A PDA detector system model has some items that can be tested by the accessory PDA
Utility. For details, refer to the Help section of the PDA Utility.

Items for automatic validation and performance check


There are different items that can be tested through automatic validation and
performance checks. Conduct testing by referring to the following.

The items that could not be tested through automatic validation can be tested through
performance checks to reduce the system check time.

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2 Validation

"Y": Can be tested, "-": Cannot be tested


Automatic Performance
Item System Check Items Ref.
Validation Check
Flow rate accuracy test - Y P.32
Solvent delivery stability test Y Y P.35
Pump Gradient concentration accuracy
Y Y P.38
test
Pressure limiter test Y Y P.41
Injection volume accuracy test - Y P.44
Autosampler Sample cooler set temperature
- Y P.47
accuracy test
Column oven set temperature
- Y P.50
accuracy test
Column Oven Temperature control accuracy
Y Y P.50
test
Gas sensor test - Y P.54
Wavelength accuracy / Lamp
Y Y P.57
energy test
Absorbance accuracy test
Y Y P.59
UV Detector (built-in filter)
Absorbance accuracy test
- Y P.61
(External standard filter)
Baseline noise/drift test Y Y P.64
Wavelength accuracy / Lamp
Y Y P.68
energy test
PDA Detector Absorbance accuracy test
- Y P.71
(External standard filter)
Baseline noise/drift test Y Y P.74
Other Parts Leak sensor test - Y P.78

To change the validation criteria, refer to "2.2.4 Calibration Tab ■ Validation


Acceptance Criteria Values Setting screen" in the Operation Guide.

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2.2 System Check

Items for other validations


The following validations do not have items on the screen.

System Check Items Ref.


Analysis Reproducibility Test P.83
Carryover Test P.92
UV Detector Absorbance Linearity Test P.94
PDA Detector Absorbance Linearity Test P.97
2
Items that can be tested using the PDA Utility (PDA detector system model)
The PDA Utility can test the following items. For operation details, refer to the Help
section of the PDA Utility.
• Wavelength accuracy test
• Light lamp intensity check
• Linearity check

2.2 System Check


Displays the history of performance checks or automatic validations, or the operating
conditions of consumable parts in a list.

Process this before analysis.

2.2.1 Processing the System Check Using the Instrument

Check the progress in the [Maintenance] screen in the [Validation] tab.

1 Press (Menu).

The [Menu] screen opens.

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2 Validation

2 Press [Maintenance].
The [Maintenance] screen is displayed.

3 Press (Validation).

The [Validation] tab is displayed.

4 Press [System Check].


The [System Check] screen is displayed.

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2.2 System Check

System Check screen

No. Name Explanation


Lists the history of the performance check or
1 System Check Result
automatic validation results.
No. Shows the line number of a system check result.
Item The name for a checked item.
Check Date The date and time at which a check was processed.
Result Value Displays the results of system checks.
Displays the results of system checks based on CHECK
Result
CRITERIA.
OK Displayed if the result of the system check is "PASS".
Displayed if the system check resulted in the use of a
* * *
consumable part exceeding the replacement guideline.
Displayed if the result of the system check is
NG
"FAILED".
- Displayed when testing is not being processed.
Pressing the arrow key turns a page.
2 Page Operation Button Above the arrow key, the current page and the total
number of pages are displayed.

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2 Validation

2.2.2 Processing the System Check Using PC, iPad, or iPhone

The user can process the check from a PC, iPad, or iPhone in the same network.
For the setting procedure of your PC, iPad, or iPhone, refer to the Operation Guide.
Operation Guide "4.9 Check the Status of the Instrument from PC", "4.10 Check the Status
of the Instrument from iPad", and "4.11 Check the Status of the Instrument from
iPhone"

1 Display the Web screen on a PC, iPad, or iPhone in the same network.

2 Click on the [System Check] tab in the


[Status Summary] area of the Web
screen.
The [System Check] screen is displayed.

3 Click on the name of the instrument or


click .

The [Login] screen opens.

4 Enter the [User ID] and [Password],


and click on [Login].
The [System Check] screen is displayed.
The previous results are displayed.

5 Click on (Execute Check).

The Confirmation screen is displayed.

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2.2 System Check

6 Click on [OK].
The [System Check Running] screen is
displayed and the system check is started.

Clicking on [Stop] while the


check is running causes the
system check to be cancelled.
The cancellation process may
take 1 to 2 minutes.
2
When the system check finishes, the [System Check Running] screen closes and the results
of system checks are displayed. The results of system checks are divided into three groups:
"System check execution and result", "Information for instrument", and "Detailed
information for system check".

n System check execution and result

Displays the overview of the system check result.

No. Display Item Explanation


Displays the name of the instrument. Clicking on this name
1 System Name
displays the instrument information.
2 Check Date The date and time at which a check was processed.
The user name that processed the system check.
User Performed
3
Check "-" is displayed in the instrument.

The comprehensive result determined from the judgment of each


Result
item.

Displayed if the results of all system checks are "PASS".

4 Displayed if the system check resulted in the use of a consumable


part exceeding the replacement guideline.

Displayed if the result of the system check is "FAILED".

- Displayed when unexecuted.

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2 Validation

n Instrument information

Displays the information for the instrument relevant to the system check.
"2.2.3 List of System Check Results" P.12

No. Display Item Explanation


Name of a unit in the instrument. Clicking on this model name
displays the advanced results of the system check.
1 Unit Name
This is displayed as "Main Unit" in the instrument.

2 Unit Model The model name of a connected instrument.


3 Version The version number of a connected instrument.
4 Serial No. The serial number of a connected instrument.
Results of system checks based on CHECK CRITERIA.
5 Result For the meanings of the displayed symbols, see "System check
execution and result" P.9 described earlier.

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2.2 System Check

n Detailed system check results

Displays the results of performance checks or automatic validation or the operating


conditions of consumable parts in a list.
"2.2.3 List of System Check Results" P.12

No. Display Item Explanation


1 Item Represents an item name.
2 Check Date The date and time at which each item's check was processed.
Result Value / Criterion
3 Displays the results of system checks and criterion values.
Value
Result of system checks based on criteria.
4 Result For the meanings of the displayed symbols, see "System check
execution and result" P.9 described earlier.

• When the [Result] field displays or , refer to the instruction manual

of each model and replace the consumable part or try to resolve the cause of
the failure.
• Powering off the instrument causes only [System Check Result] to be saved and
the data for [Unit Info] and [Details] to be erased.

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2 Validation

2.2.3 List of System Check Results

Displayed items vary between UV detector system models and PDA detector system
models.

n Shared items across units

No. Item Explanation


1 <GENERAL> -
Name of the instrument.
• LC-2030 Plus: UV detector system model without sample
cooler
2 System Name
• LC-2030C Plus: UV detector system model with sample cooler
• LC-2030C 3D Plus: PDA detector system model with sample
cooler
Date and time when the result of the system check was
3 Date Checked
recorded.
4 Serial Number Serial number of the instrument.
5 ROM Version ROM version value of the instrument.
6 IP Address IP address value of the instrument.
7 Subnet Mask Subnet mask value of the instrument.
8 Default Gateway Default gateway value of the instrument.
9 MAC Address MAC address value of the instrument.
Total time for which the instrument was used with the power
10 Total OP Time /h
ON.
11 ROM Result of checking whether the ROM status is normal.
12 RAM Result of checking whether the RAM status is normal.
Result of checking whether the autosampler HP (home
13 Position Sensor
position) sensor status is normal.
14 - -
15 <PUMP UNIT> -
16 Total Delivered /L Total value of the pumped volume.
17 Mixer volume /µL Mixer volume.
18 [Check-Consumables] -
Total value of the volume pumped from the left pump. This
19 L Seal Delivered /L becomes a criterion value for the degree of consumption for
the left plunger seal.
Total value of the volume pumped from the right pump. This
20 R Seal Delivered /L becomes a criterion value for the degree of consumption for
the right plunger seal.
21 [Calibration] -
22 Flow Compensation History of calibrations run for [Flow rate accuracy].

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2.2 System Check

No. Item Explanation


History of calibrations run for [GE composition].
23 Grad. Conc. Calib The volume of the mixer used is displayed in the Result Value
field.
Comp.Factor Correction value of a gradient concentration calibrated with
24
A/B 10% mobile phase port A at 90 % and B at 10 %.
Comp.Factor Correction value of a gradient concentration calibrated with
25
A/B 50% mobile phase port A at 50 % and B at 50 %.

26
Comp.Factor Correction value of a gradient concentration calibrated with 2
A/B 90% mobile phase port A at 10 % and B at 90 %.
Comp.Factor Correction value of a gradient concentration calibrated with
27
A/C 10% mobile phase port A at 90 % and C at 10 %.
Comp.Factor Correction value of a gradient concentration calibrated with
28
A/C 50 % mobile phase port A at 50 % and C at 50 %.
Comp.Factor Correction value of a gradient concentration calibrated with
29
A/C 90 % mobile phase port A at 10 % and C at 90 %.
Comp.Factor Correction value of a gradient concentration calibrated with
30
A/D 10 % mobile phase port A at 90 % and D: at 10 %.
Comp.Factor Correction value of a gradient concentration calibrated with
31
A/D 50 % mobile phase port A at 50 % and D at 50 %.
Comp.Factor Correction value of a gradient concentration calibrated with
32
A/D 90 % mobile phase port A at 10 % and D at 90 %.
33 [Performance Check] -
Displays the results of system checks for [Flow rate accuracy]
as follows.
[Measurement condition (mL/min)] [Measured value (%)] < / >
Flow Accuracy
34 [Check criteria (%)]
/±%
Display example: (1.5 mL) -0.17 < 0.5
* When the measured value at 1.5 mL/min was -0.17% with the
CHECK CRITERIA at ±0.5 %

Displays the results of system checks for [Pressure pulsation] as


35 Pulsation /MPa follows.
[Measured value (MPa)] < / > [Check criteria (MPa)]
Testing results with [GE composition] mobile phase ports A
and B at "90 % : 10 %", "50 % : 50 %", and "10 % : 90
Grad. Accuracy %". The results are judged as PASS only when all the CHECK
36
(A/B) (±%) CRITERIA are met.
The volume of the mixer used is displayed in the Result Value
field.
Displays the results of testing with [GE composition] mobile
Grad. Accuracy phase port A at 90 % and B at 10 % as follows.
37
(B 10 %)
[Measured value (%)] < / > [Check criteria (%)]
Displays the results of testing with [GE composition] mobile
Grad. Accuracy phase port A at 50 % and B at 50 % as follows.
38
(B 50 %)
[Measured value (%)] < / > [Check criteria (%)]

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2 Validation

No. Item Explanation


Displays the results of testing with [GE composition] mobile
Grad. Accuracy phase port A at 10 % and B at 90 % as follows.
39
(B 90 %)
[Measured value (%)] < / > [Check criteria (%)]
Testing results with [GE composition] mobile phase ports A
and C at "90 % : 10 %", "50 % : 50 %", and "10 % : 90
Grad. Accuracy %". The results are judged as PASS only when all the CHECK
40
(A/C) (±%) CRITERIA are met.
The volume of the mixer used is displayed in the Result Value
field.
Displays the results of testing with [GE composition] mobile
Grad. Accuracy phase port A at 90 % and C at 10 % as follows.
41
(C 10 %)
[Measured value (%)] < / > [Check criteria (%)]
Displays the results of testing with [GE composition] mobile
Grad. Accuracy phase port A at 50 % and C at 50 % as follows.
42
(C 50 %)
[Measured value (%)] < / > [Check criteria (%)]
Displays the results of testing with [GE composition] mobile
Grad. Accuracy phase port A at 10 % and C at 90 % as follows.
43
(C 90 %)
[Measured value (%)] < / > [Check criteria (%)]
Testing results with [GE composition] mobile phase ports A
and D at "90 % : 10 %", "50 % : 50 %", and "10 % : 90
Grad. Accuracy %". The results are judged as PASS only when all the CHECK
44
(A/D) (±%) CRITERIA are met.
The volume of the mixer used is displayed in the Result Value
field.
Displays the results of testing with [GE composition] mobile
Grad. Accuracy phase port A at 90 % and D at 10 % as follows.
45
(D 10 %)
[Measured value (%)] < / > [Check criteria (%)]
Displays the results of testing with [GE composition] mobile
Grad. Accuracy phase port A at 50 % and D at 50 % as follows.
46
(D 50 %)
[Measured value (%)] < / > [Check criteria (%)]
Displays the results of testing with [GE composition] mobile
Grad. Accuracy phase port A at 10 % and D at 90 % as follows.
47
(D 90 %)
[Measured value (%)] < / > [Check criteria (%)]
48 Pressure Limits Results of system checks for [Pressure Limits].
49 - -
50 [Autosampler] -
51 Max Inj. Vol. /µL Maximum injection volume for an injector.
52 Safety Lock Displays the status of the safetylock setting as [ON] or [OFF].
53 [Check-Consumables] -
Displays the total times the needle was used as follows.
54 Needle /time
[Total usage] < / > [Check criteria]

Needle Seal Displays the total times of injection as follows.


55
/time [Total Injection No.] < / > [Check criteria]

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2.2 System Check

No. Item Explanation


Displays the total times the high-pressure valve rotor has
H.P.V Rotor rotated as follows.
56
/rev
[Total number of revolutions] < / > [Check criteria]
Displays the total times the high-pressure valve stator has
H.P.V Stator rotated as follows.
57
/rev
[Total number of revolutions] < / > [Check criteria]
Displays the total times the low-pressure valve rotor has
58
L.P.V Rotor rotated as follows. 2
/rev
[Total number of revolutions] < / > [Check criteria]
Displays the total times the low-pressure valve stator has
L.P.V Stator rotated as follows.
59
/rev
[Total number of revolutions] < / > [Check criteria]

Sample Loop Displays the total times the sample loop was used as follows
60
/time [Total usage] < / > [Check criteria]
61 [Calibration] -
Value of the compensation factor by running calibration for
62 Inj. Accuracy Factor
[Injection Volume accuracy].
Rack Pos. Latest date and time at which rack position adjustment was
63
Adjustment processed.
Latest date and time at which injection port position
64 Inj. Port Adjustment
adjustment was processed.
65 [Performance Check] -
Displays the result of the system check for [Injection Volume
66 Inj. Accuracy /±% accuracy] as follows.
[Measured value (%)] < / > [Check criteria (%)]
67 - -
68 <OVEN UNIT> -
Displays the CMD (Column Management Device) connection as
69 CMD
[ON] (connected) or [OFF] (not connected).
70 [Check-Consumables] -
Displays the total times the flow line switching valve has
Valve (Seal) rotated as follows. If no flow line switching valve is connected,
71 [NONE] is displayed.
/rev
[Total number of revolutions] < / > [Check criteria]
72 [Calibration] -
Displays the result of the calibration run for [Temperature
accuracy] as follows.
[1st inspection temperature (°C)][1st measured value]
73 Temp.Comp.1 /°C
Display example: (25) 26.2
* When the first inspection temperature is set to 25 °C and the
external thermosensor measurement is 26.2 °C

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2 Validation

No. Item Explanation


Displays the result of the calibration run for [Temperature
accuracy] as follows.
[2nd inspection temperature (°C)][2nd measured value]
74 Temp.Comp.2 /°C
Display example: (60) 59.8
* When the second inspection temperature is set to 60 °C and the
measurement of the external thermosensor is 59.8 °C

Displays the maximum compensated LS (leak sensor) value


75 Gas Sensor Comp.
after running calibration for [Gas Sensor].
76 [Performance Check] -
Displays only the latest report on the results of system checks
for [Temperature accuracy] as follows. (The results are not
recorded for each measurement condition.)
T. Accuracy [Measurement condition (°C)][Measured value (°C)] < / > [Check
77
/±deg criteria (°C)]
Display example: (50) 0.1 < 2.0
* When the measured value at a setting of 50.0 °C is +0.1 °C with
CHECK CRITERIA at ±2.0 °C

Displays only the latest report on the results of system checks


for [Temperature control precision] as follows. (The results are
not recorded for each measurement condition.)
[Measurement condition (°C)][Measured value (°C)] < / > [Check
78 T. Stability /deg
criteria (°C)]
Display example: (50) 0.10 < 0.20
* When the measured value at a setting of 50.0 °C is +0.10 °C with
CHECK CRITERIA at ±0.20 °C

79 Gas Sensor Test Results of system checks for [Gas Sensor Test].
80 - -

n UV detector system model

No. Item Explanation


81 <DETECTOR UNIT> -
82 [Check-Consumables] -
Displays the total usage time of the D2 lamp as follows.
83 D2 Lamp Time /h
[Total usage time] < / > [Check criteria]
Displays the results of system checks for [D2 Lamp energy] or
Lamp Energy (220 the lamp check results during startup as follows.
84
nm) /mV
[Energy value] < / > [Check criteria]
85 [Calibration] -
86 Wavelength Calib. History of calibrations run for [Wavelength accuracy].
Displays the value of the compensation factor by running
87 ABS. Compensation
calibration for [Absorbance accuracy].
88 [Performance Check] -

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2.2 System Check

No. Item Explanation


Displays the results of system checks for [D2 Lamp energy] as
Lamp Energy (220 follows.
89
nm) /mV
[Measured value (mV)] < / > [Check criteria (mV)]
Results of system checks for [Wavelength accuracy] or the
90 W. Accuracy /±nm
result of the wavelength accuracy check during startup.
Displays the results of testing the [Wavelength accuracy] with
the built-in holmium oxide filter lambda max*1 as follows.
91 Wavelength1 /±nm
[Measurement condition (nm)][Measured value (nm)] < / > 2
[Check criteria (nm)]
Displays the results of testing the [Wavelength accuracy] with
the D2 lamp emission line (656.1 nm) as follows.
92 Wavelength2 /±nm
[Measurement condition (nm)][Measured value (nm)] < / >
[Check criteria (nm)]
Displays the results of system checks for [Baseline Noise/Drift]
93 Noise /mAU as follows.
[Measured value (mAU)] < / > [Check criteria (mAU)]
Displays the results of system checks for the drift in [Baseline
94 Drift /mAU/h Noise/Drift] as follows.
[Measured value (mAU/h)] < / > [Check criteria (mAU/h)]
Displays the results of system checks for the built-in filter for
95 ABS(InternalF) /±% [Absorbance accuracy] as follows.
[Measured value (%)] < / > [Check criteria (%)]
Displays the results of system checks for the external standard
96 ABS(ExternalF) /±% filter for [Absorbance accuracy] as follows.
[Measured value (%)] < / > [Check criteria (%)]
97 - -
98 <COOLER UNIT> -
99 [Calibration] -
Displays the value of the compensation factor by running
T.
100 calibration for [Cooler Temperature accuracy].
Compensation /deg
This displays values only for models with a sample cooler.
101 [Performance Check] -
Displays the results of system checks for [Cooler Temperature
accuracy] as follows.
102 T. Accuracy /±deg This displays values only for models with a sample cooler.
[Measured value (°C)] < / > [Check criteria (°C)]
103 - -
104 <OPTION> -
Displays the connection of the recycle valve as [INSTALL]
105 Recycle Valve
(connected) or [NONE] (not connected).
Displays the A/D board connection as [INSTALL] (connected) or
106 Optional Board
[NONE] (not connected).
107 - -
108 <LEAKAGE SENSOR> -

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2 Validation

No. Item Explanation


109 [Threshold Level] -
110 Pump Displays the leak sensor threshold for a pump.
AUTOSAMPLER Displays the leak sensor threshold for an autosampler (in the
111
H.P.V high-pressure valve position).
Displays the leak sensor threshold for an autosampler (in the
112 AUTOSAMPLER L.P.V
low-pressure valve position).
113 DETECTOR UNIT Displays the [Leak Sensor] threshold for a detector.
114 [Performance Check] -
115 Pump Results of system checks for [Leak Sensor] for a pump.
AUTOSAMPLER Results of system checks for [Leak Sensor] for an autosampler
116
H.P.V (in the high-pressure valve position).
Results of system checks for [Leak Sensor] for an autosampler
117 AUTOSAMPLER L.P.V
(in the low-pressure valve position).
118 DETECTOR UNIT Results of system checks for [Leak Sensor] for a detector.
119 - -
Displays the comprehensive results detailing the testing
120 Total Judgement
contents.

*1 The value varies depending on the instrument (holmium oxide filter). The lambda max wavelength
is displayed in [Measurement condition (nm)].

n PDA detector system model

No. Item Explanation


81 <DETECTOR UNIT> -
82 [Check-Consumables] -
Displays the total usage time of the D2 lamp as follows.
83 D2 Lamp Time /h
[Total usage time] < / > [Check criteria]
Displays the total usage time of the W lamp as follows.
84 W Lamp Time /h Not displayed if no W lamp is connected.
[Total usage time] < / > [Check criteria]
85 [Calibration] -
86 Wavelength Calib. History of calibrations run for [Wavelength accuracy].
Displays the value of the compensation factor by running
87 ABS. Compensation
calibration for [Absorbance accuracy].
STRAY.L ENERGY Displays stray light intensity (V) (when using a 1.2 nm slit,
88
NAR when equipped with standard flow cells).
STRAY.L BACKGND Displays standard stray light intensity (V) (when using a 1.2
89
NAR nm slit, when equipped with standard flow cells).
STRAY.L ENERGY Displays stray light intensity (V) (when using an 8 nm slit,
90
WID when equipped with standard flow cells).
STRAY.L BACKGND Displays standard stray light intensity (V) (when using an 8 nm
91
WID slit, when equipped with standard flow cells).

18 Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus


2.2 System Check

No. Item Explanation


ACCTIME D2 Displays the exposure time of the D2 lamp (when using a 1.2
92
NAR /ms nm slit).
ACCTIME D2 Displays the exposure time of the D2 lamp (when using an
93
WID /ms 8 nm slit).
Displays the exposure time of the W lamp (when using a
ACCTIME W
94 1.2 nm slit).
NAR /ms
Not displayed if no W lamp is connected.

ACCTIME W
Displays the exposure time of the W lamp (when using an 8 2
95 nm slit).
WID /ms
Not displayed if no W lamp is connected.
96 [Performance Check] -
97 Slit Check Results of system checks for the slit.
Displays the number of times the user switched between a
98 Slit Switch Times
1.2 nm slit and an 8 nm slit.
Displays the results of system checks for [Lamp energy] for the
D2 Energy D2 lamp (when using a 1.2 nm slit) as follows.
99
NAR(230nm) /V
[Measured value (V)] < / > [Check criteria (V)]
Displays the results of system checks for [Lamp energy] for the
D2 Energy D2 lamp (when using an 8 nm slit) as follows.
100
WID(230nm) /V
[Measured value (V)] < / > [Check criteria (V)]
Displays the results of system checks for [Lamp energy] for the
W Energy W lamp (when using a 1.2 nm slit) as follows. Not displayed
101 if no W lamp is connected.
NAR(600nm) / V
[Measured value (V)] < / > [Check criteria (V)]
Displays the results of system checks for [Lamp energy] for the
W Energy W lamp (when using an 8 nm slit) as follows.
102 Not displayed if no W lamp is connected.
WID(600nm) /V
[Measured value (V)] < / > [Check criteria (V)]
WAV. Acc. 656.1 Testing results with [Wavelength accuracy] at a center
103
nm(NAR) wavelength (656.1 nm) (when using a 1.2 nm slit).
WAV. Acc. 486.0 Testing results with [Wavelength accuracy] at a center
104
nm(NAR) wavelength (486.0 nm) (when using a 1.2 nm slit).
WAV. Acc. 360.8 Testing results with [Wavelength accuracy] at a center
105
nm(NAR) wavelength (360.8 nm) (when using a 1.2 nm slit).
WAV. Acc. 241.5 Testing results with [Wavelength accuracy] at a center
106
nm(NAR) wavelength (241.5 nm) (when using a 1.2 nm slit).
WAV. Acc. 656.1 Testing results with [Wavelength accuracy] at a center
107
nm(WID) wavelength (656.1 nm) (when using an 8 nm slit).
Wavelength
108 Results of system checks for [Wavelength accuracy].
Accuracy
Displays the results of system checks for [Baseline Noise/Drift]
109 Noise /mAU as follows.
[Measured value (mAU)] < / > [Check criteria (mAU)]

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2 Validation

No. Item Explanation


Displays the results of system checks for the drift in [Baseline
110 Drift /mAU/h Noise/Drift] as follows.
[Measured value (mAU/h)] < / > [Check criteria (mAU/h)]
Displays the results of system checks for the external standard
111 ABS(ExternalF) /±% filter for [Absorbance accuracy] as follows.
[Measured value (%)] < / > [Check criteria (%)]
112 - -
113 <COOLER UNIT> -
114 [Calibration] -
Displays the value of the compensation factor by running
T.
115 calibration for [Cooler Temperature accuracy].
Compensation /deg
This displays values only for models with a sample cooler.
116 [Performance Check] -
Displays the results of system checks for [Cooler Temperature
accuracy] as follows. This displays values only for models with
117 T. Accuracy /±deg a sample cooler.
[Measured value (°C)] < / > [Check criteria (°C)]
118 - -
119 <OPTION> -
Displays the A/D board connection as [INSTALL] (connected) or
120 Option Board
[NONE] (not connected).
121 - -
122 <LEAKAGE SENSOR> -
123 [Threshold Level] -
124 Pump Displays the leak sensor threshold for a pump.
AUTOSAMPLER Displays the leak sensor threshold for an autosampler (in the
125
H.P.V high-pressure valve position).
Displays the leak sensor threshold for an autosampler (in the
126 AUTOSAMPLER L.P.V
low-pressure valve position).
127 DETECTOR UNIT Displays the [Leak Sensor] threshold for a detector.
128 [Performance Check] -
129 Pump Results of system checks for [Leak Sensor] for a pump.
AUTOSAMPLER Results of system checks for [Leak Sensor] for an autosampler
130
H.P.V (in the high-pressure valve position).
Results of system checks for [Leak Sensor] for an autosampler
131 AUTOSAMPLER L.P.V
(in the low-pressure valve position).
132 DETECTOR UNIT Results of system checks for [Leak Sensor] for a detector.
133 - -
Displays the comprehensive results detailing the testing
134 Total Judgement
contents.

20 Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus


2.2 System Check

2.2.4 Saving the Results of a System Check

The user can save the results of system checks on a PC as a text file. The saved data can
be viewed and edited in an application like Excel.

• For the setting procedure of your PC, refer to the Operation Guide.
• The user cannot save the results on an iPad or iPhone.

Operation Guide "4.9 Check the Status of the Instrument from PC", 4.10 Check the Status 2
of the Instrument from iPad", and "4.11 Check the Status of the Instrument from
iPhone"

1 Display the Web screen on a PC, iPad, or iPhone in the same network.

2 Click on the [System Check] tab in the


[Status Summary] area of the Web
screen.
The [System Check] screen is displayed.

3 Click on the name of the instrument or


click .

The [Login] screen opens.

4 Enter the [User ID] and [Password],


and click on [Login].
The [System Check] screen is displayed.
The previous results are displayed.

Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus 21


2 Validation

5 Click on (File Save).

The [File Save] screen is displayed.

A filename is automatically
assigned.

6 Click on [Save to PC].


The [File Download] screen opens.

7 Click on [Save].

Do not click on [Open].


Associating the file with
another application may cause
the file to become unsaveable.
If [Open] is clicked and the
[Open With] screen is displayed,
click [Cancel] on the displayed
screen to close the screen.

The [Save as] screen opens.

8 Save the file.

22 Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus


2.3 Automatic Validation

2.3 Automatic Validation


Fully automates the testing of items that do not require manual operation in the
performance check.
This is processed during periodic inspection and when an abnormality is found in the
instrument.
"Validation items" P.3

2
During automatic validation, keep the room temperature fluctuation within the
range from +1 °C max. to -1 °C min.
The baseline may fluctuate.

Items that were not tested through automatic validation can be tested through performance
checks to improve the efficiency of the test.

2.3.1 Processing Automatic Validation

Process the automatic validation from the [Validation] tab in the [Maintenance] screen.
Prepare the following parts before the test.

Part Name Part No. Explanation


Conduct the test by replacing the mobile
Water (HPLC Grade) -
phase with water.
Used in mobile phase port A to conduct
10 mg/L Caffeine Solution -
the test in gradient analysis (GRAD.) mode.
The pressure when pumping water at 1
Resistor Tube PEEK S228-38994
mL/min is about 10 MPa.
Male Nut PEEK S228-18565 Used to connect the resistor tube.
Coupling 1.6C S228-16004-13 Used to connect the resistor tube.

1 Press (Menu) from the Main


analysis screen.
The [Menu] screen opens.

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2 Validation

2 Press [Maintenance].
The [Maintenance] screen is displayed.

3 Press (Validation).

The [Validation] tab is displayed.

4 Press [Auto Validation].


The [Auto Validation] screen is displayed.

24 Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus


2.3 Automatic Validation

Auto validation screen

No. Name Explanation


1 Operations Guide Area Displays the operating instructions and buttons.
2 Entire Procedure Displays the entire procedure. *1

Stop Cancels the operation and closes the screen.


Back Displays the previous screen.
Next Displays the next screen.
Displayed only when the results are provided after the
Finish entire procedure is finished.
Closes the screen.

Displays the current procedure position in light gray.


3 Current Procedure is displayed when the results of system checks for
a procedure are "FAILED".
*1 Testing items vary between UV detector system models and PDA detector system models.
"Validation items" P.3

5 Select the pumping mode, [ISO] or


[GRAD.], and press [Next].

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2 Validation

6 Select a mobile phase port.


• ISO mode

• GRAD. mode

Three mobile phase ports can be used


to simultaneously select up to three
combinations: A/B, A/C, and A/D.

When FCV-11AL or FCV-11ALS is connected


• The flow line set for the method is used.
• Check for loose fittings.

Operation Guide "5.1.1 Pump"

26 Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus


2.3 Automatic Validation

7 Install a resistor tube.

1 Open the column oven door.

2 Install the resistor tube (A) as


shown in the diagram.

If the resistor tube


(A) is long and
difficult to directly
connect to the
detector, adjust by
changing the
position of the
column clamp (B).

8 Close the column oven door.


System Guide "3.6.1 Attaching the Column to the Column Oven"

9 Set the mobile phase.


• ISO mode: Set water for the selected mobile phase port.
• GRAD. mode: Set a 10 mg/L caffeine solution for mobile phase port A, and set water
as mobile phase ports B, C, D, and the rinse solution port.

10 Press [Next].

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2 Validation

11 Press [Start].
Automatic validation is started.
• ISO mode
Automatically purge the selected mobile
phase ports and then conduct the tests
from [2. Wavelength] to [9. Pressure
Limits].
• GRAD. mode
Automatically purge all the mobile phase
ports and then conduct the tests from [2. Wavelength] to [10. Gradient].

Press [Stop] to cancel the test. If [OK] is pressed on the Confirmation screen, the
tests are cancelled. If [Cancel] is pressed, the tests are restarted.

When the tests finish, the result screen is displayed.

12 Check the results of system checks and


press [Exit].

Press [Logged] to record the


results. The user can check the
recorded results in the
[Validation Log] screen.

"2.6 Check the Hardware Validation


Results" P.100

28 Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus


2.3 Automatic Validation

n List of automatic validation results

Item Explanation Corrective Action upon Failure


Calibrates the "wavelength accuracy".
Results of system • UV detector"Wavelength accuracy
Wavelength checks for lamp calibration" P.129
energy.
• PDA detector"Wavelength accuracy
calibration" P.137

D2 Lamp The life of the lamp has ended. Replace with a new 2
one. If the situation persists even after replacing
with a new one, contact your Shimadzu sales office
Results of system or representative.
checks for lamp
W Lamp energy. Maintenance Guide
"5.5 Replacing the D2 Lamp"
"6.5 Replacing the D2 Lamp"
"6.6 Replacing the W Lamp (Optional Part)"

Results of system
Refer to "6.1 Failure Diagnoses and Actions" in the
Pulsation checks for pumping
System Guide.
stability.
Results of system
checks for Refer to "6.1 Failure Diagnoses and Actions" in the
Temperature
temperature control System Guide.
precision.

Results of system Calibrates the "absorbance accuracy".


Absorbance checks for UV Detector"Absorbance accuracy calibration
absorbance accuracy. (external standard filter)" P.132

Results of system
Drift checks for baseline
drifting. Refer to "6.1 Failure Diagnoses and Actions" in the
Results of system System Guide.
Noise checks for detector
baseline noise.
Results of system
Refer to "6.1 Failure Diagnoses and Actions" in the
Pressure Limits checks for the
System Guide.
pressure limiter.
Check result of Calibrates the "gradient concentration accuracy".
gradient
Gradient "Gradient concentration accuracy calibration"
concentration
accuracy. P.109

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2 Validation

2.4 Performance Check


Any individual item of a unit can be tested.
On the [Performance Check] screen, select an item and run the check while operating
the screen.

2.4.1 Displaying the Items for Performance Check

This section explains how to display each item on the [Performance Check] screen.

1 Press (Menu).

The [Menu] screen opens.

2 Press [Maintenance].
The [Maintenance] screen is displayed.

3 Press (Validation).

The [Validation] tab is displayed.

30 Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus


2.4 Performance Check

4 Press [Performance Check].


The [Performance Check] screen is
displayed.

5 Press the tab of the unit, then select


and press the item to be tested.

6 Follow the on-screen instructions to


operate.

The instructions vary depending on the item. The following is an explanation using [Flow
rate accuracy] as an example.

No. Name Explanation


1 Operations Guide Area Displays the operating instructions and buttons.
2 Entire Procedure Displays the entire procedure.
Stop Cancels the operation and closes the screen.
Back Displays the previous screen.
Next Displays the next screen.
Displayed only when the results are provided after the
Finish entire procedure is finished.
Closes the screen.
3 Current Procedure Displays the current procedure position in light gray.

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2 Validation

2.4.2 Pump

The pump is used to test the following items.

Item Explanation Ref.


Flow rate accuracy Tests the flow rate accuracy by measuring the time
P.32
test required for pumping.
Solvent delivery Monitors the pressure fluctuation and checks the status of
P.35
stability test pulsation.
Gradient
Measures the absorbance in the set concentration and tests
concentration P.38
the concentration accuracy.
accuracy test
Checks the operation of the limiter in the maximum and
Pressure limiter test P.41
minimum pressures.

n Flow rate accuracy test

A mobile phase (water) is pumped at a set flow rate. Use a stopwatch to record the
time it takes to pump 5 mL. If the recorded time is entered by the user, the flow rate
accuracy is measured.

CHECK CRITERIA
±2.00 %

Parts needed for testing

Part Name Part No. Remarks


Water (HPLC Grade) - -
The pressure when pumping water at 1
Resistor Tube PEEK S228-38994
mL/min is about 10 MPa.
Male Nut PEEK S228-18565 Used to connect the resistor tube.
Coupling 1.6C S228-16004-13 Used to connect the resistor tube.
Measuring Cylinder (Volume:
Prepare a validated one. Prepare a
10 mL) or
- measuring flask that comes with
Measuring Flask (Volume: 5
inspection results.
mL)
Stopwatch - Prepare a calibrated one.
Plumbing Tube
S228-18495-06 -
(ID 0.3 mm, OD 1/16'', 1 m)

1 Display the [Performance Check] screen.


"2.4.1 Displaying the Items for Performance Check" P.30

2 Press the [Pump] tab and then press [Flow rate accuracy].
The [Flow rate accuracy] screen is displayed.

32 Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus


2.4 Performance Check

3 Install a resistor tube.


"2.3.1 Processing Automatic Validation" P.23 step7

4 Set water as the mobile phase.

5 Press [Next].
2

6 Select a mobile phase port and flow


rate to be used, and then press [Next].

When FCV-11AL or FCV-11ALS is


connected
• The flow line set for the
method is used.
• Check for loose fittings.

Operation Guide "5.1.1 Pump"

7 Press [Start].
Purging starts and the screen showing the
purge progress is displayed.

Press [Next] to skip purging and start


pumping.

Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus 33


2 Validation

When purging has finished, or after


pressing [Next], pumping starts.

8 Check the chromatogram, and after


pumping has stabilized, press [Next].

Even if [Next] is not pressed, the


next screen is automatically
displayed after about 10
minutes.

9 Insert the end of the tube into the


validated measuring cylinder or
measuring flask, and use a stopwatch
to measure the time it takes to pump
5 mL.

To maintain a constant speed,


ensure that the tube does not
come into contact with the
water surface.

10 Enter the measured time and then


press [Next].
The results of system checks are displayed.

34 Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus


2.4 Performance Check

11 Check the results of system checks and


press [Finish].

Press [Logged] to record the


results. The user can check the
recorded results in the
[Validation Log] screen.

"2.6 Check the Hardware Validation


Results" P.100 2

12 Remove the resistor tube and close the column oven door.

n Solvent delivery stability test

This test pumps a mobile phase (water) at 1 mL/min, monitors the pressure fluctuation
for 1 minute, and then measures the maximum pressure fluctuation margin.

CHECK CRITERIA
0.20 MPa max.

Parts needed for testing

Part Name Part No. Remarks


Water (HPLC Grade) - -
The pressure when pumping water at 1
Resistor Tube PEEK S228-38994
mL/min is about 10 MPa.
Male Nut PEEK S228-18565 Used to connect the resistor tube.
Coupling 1.6C S228-16004-13 Used to connect the resistor tube.

Measurement conditions

Item Requirements
Flow Rate 1 mL/min
Oven Temperature 40 °C

1 Display the [Performance Check] screen.


"2.4.1 Displaying the Items for Performance Check" P.30

2 Press the [Pump] tab and then press [Pressure pulsation].


The [Pressure pulsation] screen is displayed.

3 Install a resistor tube.


"2.3.1 Processing Automatic Validation" P.23 step7

Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus 35


2 Validation

4 Set water as the mobile phase.

5 Close the column oven door.


System Guide "3.6.1 Attaching the Column to the Column Oven"

6 Press [Next].

7 Select a mobile phase port to be used,


and then press [Next].

When FCV-11AL or FCV-11ALS is


connected
• The flow line set for the
method is used.
• Check for loose fittings.

Operation Guide "5.1.1 Pump"

8 Press [Start].
Purging starts and the screen showing the
purge progress is displayed.

Press [Next] to skip purging and start


pumping.

36 Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus


2.4 Performance Check

When purging has finished, or after


pressing [Next], pumping starts.

9 Check the chromatogram, and after


pumping has stabilized, press [Next].

Even if [Next] is not pressed, the


next screen is automatically
displayed in 10 minutes.

Temperature control of oven


starts when pumping starts.

The screen for measuring the pressure


fluctuation is displayed.
When measurement has finished, the
message "Measurement was completed." is
displayed.

10 Press [Next].
The results of system checks are displayed.

Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus 37


2 Validation

11 Check the results of system checks and


press [Finish].

Press [Logged] to record the


results. The user can check the
recorded results in the
[Validation Log] screen.

"2.6 Check the Hardware Validation


Results" P.100

n Gradient concentration accuracy test

This test uses water and caffeine solution to perform binary low-pressure gradient.
Absorbance for each set concentration is measured and the concentration accuracy is
calculated.

CHECK CRITERIA
±2.00 % max.

Parts needed for testing

Part Name Part No. Remarks


Water (HPLC Grade) - -
10 mg/L Caffeine Solution - -
The pressure when pumping water at 1
Resistor Tube PEEK S228-38994
mL/min is about 10 MPa.
Male Nut PEEK S228-18565 Used to connect the resistor tube.
Coupling 1.6C S228-16004-13 Used to connect the resistor tube.

Measurement conditions

Item Requirements
Flow Rate 1 mL/min
Wavelength 272 nm
Oven Temperature 40 °C
A/B, A/C, or A/D 100/0 % for 10 min
A/B, A/C, or A/D 90/10 % for 5 min
Gradient Program A/B, A/C, or A/D 50/50 % for 5 min
A/B, A/C, or A/D 10/90 % for 5 min
A/B, A/C, or A/D 0/100 % for 5 min

38 Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus


2.4 Performance Check

Mobile phase ports to be used

Select the desired ports to be used from the combinations A/B, A/C, or A/D. Set
a 10 mg/L caffeine solution for mobile phase port A, and set water for mobile
phase ports B, C, D, and the rinse solution port. (Fill those ports that will not
be tested with water.)

1 Display the [Performance Check] screen.


"2.4.1 Displaying the Items for Performance Check" P.30 2

2 Press the [Pump] tab and then press [GE composition].


The [GE composition] screen is displayed.

3 Install a resistor tube.


"2.3.1 Processing Automatic Validation" P.23 step7

4 Set the mobile phase.

5 Close the column oven door.


System Guide "3.6.1 Attaching the Column to the Column Oven"

6 Press [Next].

7 Select the mobile phase ports to be


used.

When FCV-11AL or FCV-11ALS is


connected
• The flow line set for the
method is used.
• Check for loose fittings.

Operation Guide "5.1.1 Pump"

Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus 39


2 Validation

8 Press [Next].

9 Press [Start].
Rinsing starts.

The screen showing the rinse progress is


displayed.

Press [Skip] to stop the rinse


and move to the rinse for the
next port. Press [Next] to stop
the current rinse, display the
next screen, and start the
system equilibration.

After all ports have been rinsed, the next


screen is displayed and the oven
temperature and detector equilibrations
start.

Press [Next] to stop the current


system equilibration and display
the next screen.

When the oven temperature and detector


equilibrations have finished, the next
screen is displayed and absorbance is
measured for each concentration.
After measurement has finished, the [Next]
button is activated.

40 Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus


2.4 Performance Check

10 Press [Next].
The results of system checks are displayed.

11 Check the results of system checks and


press [Finish].

Press [Logged] to record the


results. The user can check the
recorded results in the
[Validation Log] screen.

"2.6 Check the Hardware Validation


Results" P.100

n Pressure limiter test

This test first pumps a mobile phase (water) at 1 mL/min and then measures the pressure
when pumping stabilizes. Let this value be "X". After measurement, the upper limit
limiter is automatically set to (X+1) MPa, and the lower limit limiter is set to (X-1) MPa.
After that, the flow rate is changed to 1.5 mL/min and then 0.5 mL/min sequentially,
and check that the upper limit limiter and lower limit limiter are checked for normal
operation.

CHECK CRITERIA
The upper limit limiter and the lower limit limiter should operate normally.

Parts needed for testing

Part Name Part No. Remarks


Water (HPLC Grade) - -
The pressure when pumping water at
Resistor Tube PEEK S228-38994
1 mL/min is about 10 MPa.
Male Nut PEEK S228-18565 Used to connect the resistor tube.
Coupling 1.6C S228-16004-13 Used to connect the resistor tube.

Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus 41


2 Validation

1 Display the [Performance Check] screen.


"2.4.1 Displaying the Items for Performance Check" P.30

2 Press the [Pump] tab and then press [Pressure Limits].


The [Pressure Limits] screen is displayed.

3 Install a resistor tube.


"2.3.1 Processing Automatic Validation" P.23 step7

4 Set the mobile phase.

5 Close the column oven door.


System Guide "3.6.1 Attaching the Column to the Column Oven"

6 Press [Next].

Check that the drain valve is


tightened.

7 Select the mobile phase ports to be


used.

When FCV-11AL or FCV-11ALS is


connected
• The flow line set for the
method is used.
• Check for loose fittings.

Operation Guide "5.1.1 Pump"

42 Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus


2.4 Performance Check

8 Press [Start].
Measurement starts and the screen
showing the measurement progress is
displayed.
When measurement finishes, the check
results are displayed.

9 Check the results of system checks and


press [Exit].

Press [Logged] to record the


results. The user can check the
recorded results in the
[Validation Log] screen.

"2.6 Check the Hardware Validation


Results" P.100

Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus 43


2 Validation

2.4.3 Autosampler

The autosampler is used to test the following items.

Item Explanation Ref.


Measures the difference in mass between before and
Injection volume accuracy
after injection of the sample to test the injection P.44
test
volume accuracy.
Tests the temperature accuracy of the sample cooler
Sample cooler set
based on the difference from the temperature P.47
temperature accuracy test
measured by an external thermosensor.

n Injection volume accuracy test

This test uses a 1.5 mL sample vial containing a sample (water) to measure the mass
before and after 10 repeated injections of 50 µL. Based on the difference in mass, the
injection volume accuracy is calculated.

CHECK CRITERIA
±1.3 % max.

Parts needed for testing

Part Name Part No. Remarks


1.5 mL Sample Vial S228-15652-92 -
Water (HPLC Grade) - -
Balance - Use a validated and calibrated one.
Sample Rack S228-55735-41 Standard accessory.
1.5 mL Sample Vial Plate S228-50830-92
Place the 1.5 mL sample vial on the
or
plate.
1.5 mL Sample Vial Metal Plate S228-61615-42

• For models with a sample cooler, turn OFF the temperature control of the
sample cooler to prevent condensation within the autosampler. For the setting
procedure, refer to "2.1 Analysis Screen" in the Operation Guide.
• Before testing, thoroughly flush the mobile phase port to be used and the
rinse solution with water (HPLC grade).
• When the optional sample loop for 50 µL injections is used, set the sample
aspiration rate to 5 µL/sec or less.

1 Display the [Performance Check] screen.


"2.4.1 Displaying the Items for Performance Check" P.30

2 Press the [Autosampler] tab and then press [Injection Volume accuracy].
The [Injection Volume accuracy] screen is displayed.

44 Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus


2.4 Performance Check

3 Put 1 mL of water into the 1.5 mL


sample vial (A), and measure the mass
using a balance.

4 Enter the measured mass.

Enter the value to the 4th


decimal place.

5 Set the weighed sample vial


containing water and fitted with a
silicone rubber septum on position 1 of
plate 1 (left side) in rack L.

6 Press [Next].

Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus 45


2 Validation

7 Select a mobile phase port to be used,


and then press [Next].
The screen showing the operation progress
is displayed.

When FCV-11AL or FCV-11ALS is


connected
• The flow line set for the
method is used.
• Check for loose fittings.

Operation Guide "5.1.1 Pump"

When injection is completed, the next


screen is displayed.

8 Remove the 1.5 mL sample vial (A) and


measure the mass using a balance.

9 Enter the measured mass and then


press [Next].

Enter the value to the 4th


decimal place.

The results of system checks are displayed.

46 Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus


2.4 Performance Check

10 Check the results of system checks and


press [Finish].

Press [Logged] to record the


results. The user can check the
recorded results in the
[Validation Log] screen.

"2.6 Check the Hardware Validation


Results" P.100 2

n Sample cooler set temperature accuracy test

This test uses an external thermosensor to measure the temperature of the sample cooler.
If the measured temperature is entered by the user, the temperature accuracy of the
sample cooler is calculated. The set value of the sample cooler is "4 °C".

CHECK CRITERIA
±3.0 °C max.

It is not possible to cool to 1 °C or below.

Parts needed for testing

Part Name Part No. Remarks


Water (HPLC Grade) - -
External Thermosensor - Use a calibrated one.
Sample Rack S228-55735-41 Standard accessory.
Place the 1.5 mL sample vial on the
1.5 mL Sample Vial Metal Plate S228-61615-42
plate.
1.5 mL Sample Vial S228-15652-92 -

1 Display the [Performance Check] screen.


"2.4.1 Displaying the Items for Performance Check" P.30

2 Press the [Autosampler] tab and then press [Cooler Temperature accuracy].
The [Cooler Temperature accuracy] screen is displayed.

Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus 47


2 Validation

3 Fit the external thermosensor to the


sample vial.
1 Put 1 mL of water into the 1.5 mL
sample vial (A), and insert the
external thermosensor (B) through
the septum until it touches the
vial bottom.

2 Set the sample vial with the


external thermosensor attached
into position 27 on the 1.5 mL
sample vial Metal plate.

Check that, when


setting it, the sample
vial is correctly fitted
to the plate.

3 Remove the right and left sample


racks. Set the plate fitted with
the external thermosensor on
plate 3 of rack R. (Only set plates
on the left front, left back, and
right back sides.)

48 Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus


2.4 Performance Check

4 Insert the sample racks.

When inserting the


sample racks, ensure
that the external
thermosensor cable
does not get pinched
inside the
instrument. If the
cable is caught, it
will prevent the 2
sample racks from
being fully inserted,
resulting in
inaccurate testing.

4 Press [Next].
The sample cooler starts temperature
control at a set temperature of 4 °C.

When the temperature of the sample


cooler stabilizes, the next screen is
displayed.

If [Next] is pressed, the next


screen is displayed even if the
temperature of the sample
cooler is not stable.

5 Read the temperature of the sample


vial's external thermosensor.

6 Enter the read temperature and then


press [Next].
The results of system checks are displayed.

Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus 49


2 Validation

7 Check the results of system checks and


press [Finish].

Press [Logged] to record the


results. The user can check the
recorded results in the
[Validation Log] screen.

"2.6 Check the Hardware Validation


Results" P.100

2.4.4 Column Oven

The column oven is used to test the following items.

Item Explanation Ref.


Column oven set
Tests the temperature accuracy of the column oven based
temperature
on the difference between the temperature measured by
accuracy / P.50
an external thermosensor and the actual temperature. The
Temperature control
temperature control precision is also tested.
accuracy test
After the injection of ethanol into the column oven, checks
Gas sensor test P.54
the operation of the sensor.

n Column oven set temperature accuracy / Temperature control accuracy test

This test lets the user select the oven temperature from among 40 °C, 50 °C, and room
temperature -5 °C, and, once the temperature stabilized, measure the temperature of the
column holder using an external thermosensor. The oven temperature accuracy and
fluctuation are measured from the automatic measurement result and the measured
temperature of the temperature fluctuation.

The user can check the room temperature in the monitor value display screen.

Operation Guide "2.1.6 Chromatograph Display Tab"

CHECK CRITERIA
• Temperature accuracy: ±2.0 °C max.

• Temperature control precision: dT 0.20 °C max.

Parts needed for testing

Part Name Part No. Remarks


External thermosensor - Use a calibrated one.
Used to attach the external
Aluminum Tape -
thermosensor to the column holder.

50 Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus


2.4 Performance Check

1 Display the [Performance Check] screen.


"2.4.1 Displaying the Items for Performance Check" P.30

2 Press the [Oven] tab and then press [Oven Temperature accuracy/fluctuation].
The [Oven Temperature accuracy/fluctuation] screen is displayed.

3 Attach the external thermosensor.


2
1 Open the column oven door.

2 Use 1 cm wide aluminum tape (B)


to attach the end of the external
thermosensor (A) to the column
holder as shown in the diagram.

If necessary, tape at
two or more points
to ensure that the
external
thermosensor will
not fall off the
column holder.

3 Close the column oven door.


System Guide "3.6.1
Attaching the Column to
the Column Oven"

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2 Validation

4 Press [Next].

5 Select a testing temperature.

Selecting [Room Temp. -5 °C]


sets the temperature 5 °C lower
than the room temperature
measured by the room
temperature sensor of the
instrument.

6 Press [Next].

The following screen is displayed, and


temperature control starts.

52 Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus


2.4 Performance Check

When the temperature stabilizes, the


following screen is automatically displayed,
starting measurement of the temperature
fluctuation margin.
After measurement has finished, the [Next]
button is activated.

7 Press [Next].
The next screen is displayed.

8 Enter the temperature measured using


the external thermosensor, and then
press [Next].
The results of system checks are displayed.

9 Check the results of system checks and


press [Exit].

Press [Logged] to record the


results. The user can check the
recorded results in the
[Validation Log] screen.

"2.6 Check the Hardware Validation


Results" P.100

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2 Validation

n Gas sensor test

This test checks whether the gas sensor can detect organic solvent in the column oven.

Parts needed for testing

Part Name Part No. Remarks


Use one with a volume of 100 µL min.,
Syringe - a needle length of about 50 mm and a
needle outer diameter of 0.5 mm min.
Ethanol - Use about 100 µL.

Measurement conditions

Item Requirements
Oven Temperature 50 °C

If gas sensor detection is checked with condensation water generated in the


column oven due to temperature cooling control, the gas sensor may mistakenly
detect a fluid leak. Therefore, dry the inside of the oven naturally before
checking the gas sensor detection.

1 Display the [Performance Check] screen.


"2.4.1 Displaying the Items for Performance Check" P.30

2 Press the [Oven] tab and then press [Gas Sensor].


The [Gas Sensor] screen is displayed.

3 Check the tubing.

1 Open the column oven door and


check for column or tubing
installation problems.

2 Close the column oven door.


System Guide "3.6.1
Attaching the Column to
the Column Oven"

54 Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus


2.4 Performance Check

4 Press [Next].

2
The following screen is displayed, and
temperature control starts.
When the temperature stabilizes, the
message "Press "Next" button to start
measurement." is displayed.
Press [Next].

5 Use a syringe (B) to inject ethanol into


the column oven.
Use a syringe (B) to inject about 100 µL of
ethanol into the interior of the column
oven through the tubing hole slit (A).
When measurement has finished, the
message "Measurement was completed.
Press the "Next" button." is displayed.

Check that the needle of the


syringe is inserted all the way
and then inject ethanol.

6 Press [Next].
The results of system checks are displayed.

Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus 55


2 Validation

7 Check the results of system checks and


press [Exit].

Press [Logged] to record the


results. The user can check the
recorded results in the
[Validation Log] screen.

"2.6 Check the Hardware Validation


Results" P.100

2.4.5 UV Detector

The UV detector is used to test the following items.

Item Explanation Ref.


Wavelength Tests the wavelength accuracy by measuring the difference
accuracy / Lamp between the set wavelength and the measured P.57
energy test wavelength. The D2 lamp energy is also checked.
Absorbance accuracy Tests the absorbance accuracy by measuring the difference
test in absorbance between the ON and OFF statuses of the P.59
(Built-In Filter) built-in filter.
Absorbance accuracy
Tests the accuracy by measuring the difference in
test
absorbance between when an external standard filter is P.61
(External Standard
mounted and when unmounted.
Filter)
Baseline noise/drift Checks the baseline noise and drift intensities after the
P.64
test baseline stabilizes.

D2 lamp and baseline noise


As the operating time elapses, the D2 lamp will gradually decrease in lamp intensity and
increase in baseline noise.
Normally, the amplitude of noise (N) is inversely proportional to the square root of the
lamp energy (E). Roughly speaking, if the lamp energy becomes half, the noise will be
about 1.4 times.
For daily analysis, a target peak whose S/N ratio is higher than a certain value may be
usable even if its lamp intensity is small. In addition, the amplitude of noise does not
affect the accuracy of analysis data. The lamp intensity and noise/drift criteria should be
set depending on the sensitivity of the lamp energy and the accuracy of the data required
by each analysis. Normally, 10 is the criterion value for the S/N ratio.

56 Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus


2.4 Performance Check

n Wavelength accuracy / Lamp energy test

This test is used to measure the difference between the preset wavelength and the
measured wavelength. The lambda max*1 of the built-in filter (holmium oxide filter) and
the 656.1 nm emission line of the D2 lamp (deuterium (D2) lamp) are measured, and the
instrument automatically assesses whether both are within ±1.0 nm.
The D2 lamp energy is also checked.
*1 The value varies depending on the instrument (holmium oxide filter). For the lambda max
wavelength, see the criteria (240.4 nm to 241.7 nm) in the system check result screen of Procedure
7.
2
CHECK CRITERIA
• Wavelength accuracy: ±1.0 nm max.

• D2 lamp intensity (220 nm): 400 mV min.

Parts needed for testing

Part Name Part No. Remarks


Water (HPLC Grade) - -

1 Display the [Performance Check] screen.


"2.4.1 Displaying the Items for Performance Check" P.30

2 Press the [Detector] tab and then press [Wavelength accuracy/Lamp energy].
The [Wavelength accuracy/Lamp energy] screen is displayed.

3 Select a mobile phase port to be used,


and then press [Next].
The next screen is displayed.

When FCV-11AL or FCV-11ALS is


connected
• The flow line set for the
method is used.
• Check for loose fittings.

Operation Guide "5.1.1 Pump"

Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus 57


2 Validation

4 Press [Start].
Purging starts and the screen showing the
purge progress is displayed.

Press [Next] to skip purging and start


measurement.

When purging has finished, or after


pressing [Next], measurement starts.

5 Wait for measurement to finish.


When measurement has finished, the
message "Measurement was completed." is
displayed.

6 Press [Next].
The results of system checks are displayed.

58 Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus


2.4 Performance Check

7 Check the results of system checks and


press [Exit].

Press [Logged] to record the


results. The user can check the
recorded results in the
[Validation Log] screen.

"2.6 Check the Hardware Validation


Results" P.100 2

n Absorbance accuracy test (built-in filter)

This test measures absorbances when the built-in filter is OFF and ON and uses the
difference between these two absorbances and the absolute absorbance inherent to the
built-in filter in order to calculate the absorbance accuracy of the detector.

CHECK CRITERIA
±3.0 % max.

Parts needed for testing

Part Name Part No. Remarks


Water (HPLC Grade) - -

1 Display the [Performance Check] screen.


"2.4.1 Displaying the Items for Performance Check" P.30

2 Press the [Detector] tab and then press [Absorbance accuracy].


The [Absorbance accuracy] screen is displayed.

3 For a testing filter, select [Built-in


filter].

Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus 59


2 Validation

4 Select a mobile phase port to be used, and then press [Next].


The next screen is displayed.

When FCV-11AL or FCV-11ALS is connected


• The flow line set for the method is used.
• Check for loose fittings.

Operation Guide "5.1.1 Pump"

5 Press [Start].
Purging starts and the screen showing the
purge progress is displayed.

Press [Next] to skip purging and start


measurement.

When purging has finished, or after


pressing [Next], measurement starts.

6 Wait for measurement to finish.


When measurement has finished, the
message "Measurement was completed." is
displayed.

60 Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus


2.4 Performance Check

7 Press [Next].
The results of system checks are displayed.

8 Check the results of system checks and


press [Exit].

Press [Logged] to record the


results. The user can check the
recorded results in the
[Validation Log] screen.

"2.6 Check the Hardware Validation


Results" P.100

n Absorbance accuracy test (external standard filter)

This test measures the absorbance twice, once with the external standard filter not
mounted and once with the filter mounted. Based on the difference and the absolute
absorbance of the external standard filter, the absorbance accuracy of the detector is
calculated.

CHECK CRITERIA
±3.0 % max.

Parts needed for testing

Part Name Part No. Explanation


Absorbance Measurement Cell S228-57174-41 -
Standard Absorbance Filter
- JQA-manufactured JCRM230A
(External Standard Filter)
Water (HPLC Grade) - -

1 Display the [Performance Check] screen.


"2.4.1 Displaying the Items for Performance Check" P.30

2 Press the [Detector] tab and then press [Absorbance accuracy].


The [Absorbance accuracy] screen is displayed.

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2 Validation

3 For a testing filter, select [Standard


Filter].

4 Enter the wavelength and actual


absorbance value of the standard
absorbance filter (external standard
filter), and then press [Next].
The next screen is displayed.

5 Attach the absorbance measurement


cell.
1 Open the controller panel.

2 Remove the flow cell and attach


the absorbance measurement cell
(A).

Do not mount the


standard absorbance
filter (external
standard filter) at
this stage in the
procedure. Mount
the filter in
Procedure 7.

3 Close the controller panel.

6 Press [Next].
Absorbance (background absorbance) with
the filter not mounted is adjusted to "0"
and the [Filter Installing] screen is
displayed.

62 Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus


2.4 Performance Check

7 Install the standard absorbance filter


(external standard filter).
1 Open the controller panel.

2 Install the standard absorbance


filter (external standard filter) (A)
to the absorbance measurement
cell.

3 Close the controller panel.

8 Press [Start].
The next screen is displayed, starting
measurement of absorbance using the
standard absorbance filter (external
standard filter).

When measurement has finished, the


message "Measurement was completed." is
displayed.

Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus 63


2 Validation

9 Press [Next].
The results of system checks are displayed.

10 Check the results of system checks and


press [Exit].

Press [Logged] to record the


results. The user can check the
recorded results in the
[Validation Log] screen.

"2.6 Check the Hardware Validation


Results" P.100

n Baseline noise/drift test

This test measures the baseline noise and drift for 15 minutes after the baseline stabilizes.

CHECK CRITERIA
• Drift (when solvent is being delivered): 0.60 mAU/h
• Noise (when solvent is being delivered): 0.0200 mAU

Parts needed for testing

Part Name Part No. Remarks


Water (HPLC Grade) - -
The pressure when pumping water at
Resistor Tube PEEK S228-38994
1 mL/min is about 10 MPa.
Male Nut PEEK S228-18565 Used to connect the resistor tube.
Coupling 1.6C S228-16004-13 Used to connect the resistor tube.

Measurement conditions

Item Requirements
Flow Rate 1 mL/min
Oven Temperature 40 °C
Wavelength 250 nm
Response 2 sec
Cell Temperature 40 °C

64 Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus


2.4 Performance Check

1 Display the [Performance Check] screen.


"2.4.1 Displaying the Items for Performance Check" P.30

2 Press the [Detector] tab and then press [Baseline Noise&Drift].


The [Baseline Noise&Drift] screen is displayed.

3 Install a resistor tube.


2
"2.3.1 Processing Automatic Validation" P.23 step7

4 Set the mobile phase.

5 Close the column oven door.


System Guide "3.6.1 Attaching the Column to the Column Oven"

6 Press [Next].
The next screen is displayed.

7 Select a mobile phase port to be used,


and then press [Next].
The next screen is displayed.

When FCV-11AL or FCV-11ALS is


connected
• The flow line set for the
method is used.
• Check for loose fittings.

Operation Guide "5.1.1 Pump"

Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus 65


2 Validation

8 Press [Start].
Purging starts and the screen showing the
purge progress is displayed.

Press [Next] to skip purging and start


pumping.

After the purge finishes, the following


screen is displayed, and pumping starts.

Wait for 60 minutes until the instrument


becomes stable.

Press [Next] to skip waiting for


stabilization and start measurement.
Immediately after the lamp is turned
on or the temperature control starts,
the instrument is not stable, and the
testing may not perform correctly.

Start measuring the baseline drift and


noise.
When measurement has finished, the
message "Measurement was completed.
Press the "Next" button." is displayed.

66 Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus


2.4 Performance Check

9 Press [Next].
The results of system checks are displayed.

10 Check the results of system checks and


press [Exit].

Press [Logged] to record the


results. The user can check the
recorded results in the
[Validation Log] screen.

"2.6 Check the Hardware Validation


Results" P.100

Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus 67


2 Validation

2.4.6 PDA Detector

The PDA detector is used to test the following items.

Item Explanation Ref.


Tests the wavelength accuracy by measuring
Wavelength accuracy / Lamp energy the difference between the set wavelength
P.68
test and the measured wavelength. The D2 lamp
energy is also checked.
Tests the accuracy by measuring the
Absorbance accuracy test difference in absorbance between when an
P.71
(External Standard Filter) external standard filter is mounted and
when unmounted.
Checks the baseline noise and drift
Baseline noise/drift test P.74
intensities after the baseline stabilizes

"Wavelength accuracy / D2 lamp and W lamp (when an optional W lamp is


connected) intensity check" can also be performed in "PDA Utility".
For operation details, refer to the Help section of the PDA Utility.

"4.3 Wavelength Accuracy Testing," "4.4 Checking the Intensity of the Light" in the Help
section of the PDA Utility

n Wavelength accuracy / Lamp energy test

This test is used to measure the difference between the preset wavelength and the
measured wavelength. Using the emission line of the D2 lamp (deuterium (D2) lamp) and
the built-in filter (holmium oxide filter) lambda max, the wavelength accuracy is checked.
The D2 lamp intensity and W lamp intensity (when an optional W lamp is connected) is
also checked.

Wavelengths checked through testing

Item Wavelength
241.5 nm
Holmium Oxide Filter Lambda Max
360.8 nm
486.0 nm
Emission Lines of the D2 Lamp
656.1 nm*

The above wavelengths assume that a 1.2 nm slit is used. When an 8 nm slit is
used, only the wavelength marked with * is checked. (The 1.2 nm and 8 nm slits
are automatically changed.)

CHECK CRITERIA
• Wavelength accuracy: ±1.0 nm max.

• Maximum D2 lamp intensity (200 nm to 260 nm): 2.5 V min.


• Maximum W lamp intensity (when an optional W lamp is connected) (400 nm to 800
nm): 2.5 V min.

68 Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus


2.4 Performance Check

Parts needed for testing

Part Name Part No. Remarks


Water (HPLC Grade) - -

1 Display the [Performance Check] screen.


"2.4.1 Displaying the Items for Performance Check" P.30

2
2 Press the [Detector] tab and then press [Wavelength accuracy/Lamp energy].
The [Wavelength accuracy/Lamp energy] screen is displayed.

3 Select a mobile phase port to be used,


and then press [Next].
The next screen is displayed.

When FCV-11AL or FCV-11ALS is


connected
• The flow line set for the
method is used.
• Check for loose fittings.

Operation Guide "5.1.1 Pump"

4 Press [Start].
Purging starts and the screen showing the
purge progress is displayed.

Press [Next] to skip purging and start


measurement.

When purging has finished, or after


pressing [Next], measurement starts.

Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus 69


2 Validation

5 Wait for measurement to finish.


When measurement has finished, the
message "Measurement was completed." is
displayed.

6 Press [Next].
The results of system checks are displayed.

7 Check the results of system checks and


press [Exit].

Press [Logged] to record the


results. The user can check the
recorded results in the
[Validation Log] screen.

"2.6 Check the Hardware Validation


Results" P.100

70 Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus


2.4 Performance Check

n Absorbance accuracy test (external standard filter)

This test measures the absorbance twice, once with the external standard filter not
mounted and once with the filter mounted. Based on the difference and the absolute
absorbance of the external standard filter, the absorbance accuracy of the detector is
calculated.

Wait for 90 minutes or more after turning on a lamp. Because absorbance is not
stabilized immediately after lamp lighting.
2
CHECK CRITERIA
±3.0 %max.

Parts needed for testing

Part Name Part No. Remarks


Absorbance Measurement Cell S228-57174-41 S228-35011-91 can also be used.
Standard Absorbance Filter
- JQA-manufactured JCRM230A
(External Standard Filter)
Water (HPLC Grade) - -

1 Display the [Performance Check] screen.


"2.4.1 Displaying the Items for Performance Check" P.30

2 Press the [Detector] tab and then press [Absorbance accuracy].


The [Absorbance accuracy] screen is displayed.

3 Enter the wavelength and actual


absorbance value of the standard
absorbance filter (external standard
filter), and then press [Next].
The next screen is displayed.

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2 Validation

4 Attach the absorbance measurement


cell.
1 Open the controller panel.

2 Remove the flow cell and attach


the absorbance measurement cell
(A).

Do not mount the


standard absorbance
filter (external
standard filter) at
this stage in the
procedure. Mount
the filter in
Procedure 7.

3 Close the controller panel.

5 Press [Next].
Absorbance (background absorbance) with
the filter not mounted is adjusted to "0"
and the [Filter Installing] screen is
displayed.

72 Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus


2.4 Performance Check

6 Install the standard absorbance filter


(external standard filter).
1 Open the controller panel.

2 Install the standard absorbance


filter (external standard filter) (A)
to the absorbance measurement
cell.

3 Close the controller panel.

7 Press [Start].
The next screen is displayed, starting
measurement of absorbance using the
standard absorbance filter (external
standard filter).

When measurement has finished, the


message "Measurement was completed." is
displayed.

Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus 73


2 Validation

8 Press [Next].
The results of system checks are displayed.

9 Check the results of system checks and


press [Finish].

Press [Logged] to record the


results. The user can check the
recorded results in the
[Validation Log] screen.

"2.6 Check the Hardware Validation


Results" P.100

After the check, remove the standard absorbance filter (external standard filter)
immediately. The filter may be damaged.

n Baseline noise/drift test

This test measures the baseline noise and drift for 15 minutes after the baseline stabilizes.

Criteria
• Drift (when solvent is being delivered): 2.00 mAU/h max.
• Noise (when solvent is being delivered): 0.0500 mAU

Parts needed for testing

Part Name Part No. Remarks


Water (HPLC Grade) - -
The pressure when pumping water at 1
Resistor Tube PEEK S228-38994
mL/min is about 10 MPa.
Male Nut PEEK S228-18565 Used to connect the resistor tube.
Coupling 1.6C S228-16004-13 Used to connect the resistor tube.

74 Prominence-i LC-2030 Plus LC-2030C Plus LC-2030C 3D Plus


2.4 Performance Check

Measurement conditions

Item Requirements
Flow Rate 1 mL/min
Oven Temperature 40 °C
Wavelength 250 nm
Bandwidth ±4 nm
Reference Correction OFF
Slit Width 8 nm
2
Sampling 2 sec
Time Constant 2 sec
Cell Temperature 40 °C

1 Display the [Performance Check] screen.


"2.4.1 Displaying the Items for Performance Check" P.30

2 Press the [Detector] tab and then press [Baseline Noise&Drift].


The [Baseline Noise&Drift] screen is displayed.

3 Install a resistor tube.


"2.3.1 Processing Automatic Validation" P.23 step7

4 Set the mobile phase.

5 Close the column oven door.


System Guide "3.6.1 Attaching the Column to the Column Oven"

6 Press [Next].
The next screen is displayed.

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2 Validation

7 Select a mobile phase port to be used,


and then press [Next].
The next screen is displayed.

When FCV-11AL or FCV-11ALS is


connected
• The flow line set for the
method is used.
• Check for loose fittings.

Operation Guide "5.1.1 Pump"

8 Press [Start].
Purging starts and the screen showing the
purge progress is displayed.

Press [Next] to skip purging and start


pumping.

After the purge finishes, the following


screen is displayed, and pumping starts.

Wait for 90 minutes until the instrument


becomes stable.

Press [Next] to skip waiting for


stabilization and start measurement.
Immediately after the lamp is turned
on or the temperature control starts,
the instrument is not stable, and the
testing may not perform correctly.

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2.4 Performance Check

Start measuring the baseline noise and


drift.
When measurement has finished, the
message "Measurement was completed.
Press the "Next" button." is displayed.

9 Press [Next].
The results of system checks are displayed.

10 Check the results of system checks and


press [Exit].

Press [Logged] to record the


results. The user can check the
recorded results in the
[Validation Log] screen.

"2.6 Check the Hardware Validation


Results" P.100

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2 Validation

2.4.7 Others

The "Others" section is used to test the following items.

Item Explanation Ref.


Leak sensor test Checks the leak sensor sensitivity of each unit. P.78

n Leak sensor test

There are cases where the leak sensor sensitivity is not optimal due to the influence of
changes in operating environment.
Check the leak sensor sensitivities of the detector, pump, and autosampler (HPV
(high-pressure valve) and LPV (low-pressure valve)).

Parts needed for testing

Part Name Part No. Remarks


Water (HPLC Grade) - -
Syringe - -

1 Display the [Performance Check] screen.


"2.4.1 Displaying the Items for Performance Check" P.30

2 Press the [Other] tab and then press [Leak Sensor].


The [Leak Sensor] screen is displayed.

3 Select a leak sensor to be tested.


Multiple sensors can be selected for
testing.

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2.4 Performance Check

4 Pour water under the leak sensor to be tested.

UV and PDA detectors

1 Open the controller panel.

2 Use the syringe to soak the leak


sensor (A) with water.

3 Close the controller panel.

Pump

1 Open the controller panel.

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2 Validation

2 Use the syringe to soak the leak


sensor (A) with water.

3 Close the controller panel.

Autosampler (HPV)

1 Remove the inserted sample racks


(A) and open the panels and
doors (B) as shown in the
diagram.

2 Remove the 4 knurled screws (A)


and pull the front panel (B)
forward.

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2.4 Performance Check

3 Use the syringe to soak the leak


sensor (A) with water.

4 With the bundle of tubes (C)


passed through the gap (c) on the
left side of the front panel, align
the groove (b) on the bottom of
the front panel with the
protrusion (a) on the main body
frame and insert the front panel.
Attach the 4 knurled screws (A) to
attach the front panel (B).

If the front panel (B)


is not attached
correctly,
condensation may
settle within the
sample cooler.

5 Attach the removed sample racks


(A) and close all the doors (B).

For models with a


sample cooler, do
not conduct testing
without fitting the
front panel.
Condensation will
settle inside the
autosampler. When
testing, turn OFF the
sample cooler
temperature control.
Refer to "2.1
Analysis Screen" in
the Operation Guide.

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2 Validation

Autosampler (LPV)

1 Open the valve mount door.

2 Use the syringe to soak the leak


sensor (A) with water.

3 Close the valve mount door.

5 Press [Start].
Start the measurement.

When measurement has finished, the


message "Measurement was completed." is
displayed.

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2.5 Other Validations

6 Press [Next].
The results of system checks are displayed.

7 Wipe the water off the tested leak


sensor.

8 Check the results of system checks and


press [Finish].

Press [Logged] to record the


results. The user can check the
recorded results in the
[Validation Log] screen.

"2.6 Check the Hardware Validation


Results" P.100

2.5 Other Validations


This section explains the validations that are not included as items in the [Performance
Check] screen.
Validation is performed using LabSolutions (dedicated workstation). For details on how
to operate, refer to the Help section of LabSolutions.

2.5.1 Analysis Reproducibility Test

The Analysis Reproducibility Test is a test to judge whether the cause of any trouble lies
in the instrument or if the problem is inherent to each analysis.
If trouble occurs, first conduct this test, and, depending on the result, take action as
follows.

The instrument is normal. Because the cause of the trouble


When CHECK CRITERIA Are
is considered to lie somewhere else, check if the analysis
Met
conditions are proper.
Each unit should be inspected.
When CHECK CRITERIA Are
• "2.3 Automatic Validation" P.23
Not Met
• "2.4 Performance Check" P.30

In addition, conduct this test during a periodic inspection, too, in order to check if the
instrument operates normally.

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2 Validation

n Isocratic analysis

This test checks if reproducible data can be obtained from an actual analysis.
This is determined from the coefficients of variation for the peak area and the retention
time of each peak.

Criteria
• "Retention time" coefficient of variation: 0.5 % max.
• "Peak area" coefficient of variation: 1.0 % max.

Parts needed for testing

Part Name Part No. Remarks


Column - Either the column to the left or an
equivalent ODS column can be used.
Shim-Pack XR-ODS S228-41606-92 (Particle diameter: 2.2 µm, Inside diameter:
3.0 mm, Length: 50 mm)
Mixtures of water and methanol (4/1, v/v).
Mobile Phase - Use HPLC-grade water (distilled water) and
methanol.
20 mg/L caffeine solution (contained in Part
No. S228-45727-41: Caffeine set (5
concentrations)) or one that has been
adjusted using the following procedure.
1 Weigh out 20 mg of caffeine, put it in
a measuring flask with a capacity of
Sample -
100 mL, and dissolve the caffeine in
water so that the total volume becomes
100 mL.
2 Put that 1 mL in a measuring flask with
a capacity of 10 mL, and add water so
that the total volume becomes 10 mL.
Water (HPLC Grade) - -
2-Propanol (HPLC grade) - -
1.5 mL Sample Vial S228-15652-92 Use 2 sample vials.
1.5 mL Sample Vial Plate
S228-50830-92
or
or Place the 1.5 mL sample vials on this plate.
1.5mL Sample Vial Metal
S228-61615-42
Plate
LabSolutions - Data processing instrument

Before starting the test, rinse the flow line depending on the operating condition
of the instrument.

1 Instead of the column, connect the column inlet and outlet tubes to the PEEK
tubing using a coupling.

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2.5 Other Validations

2 Set the mobile phase and rinse the flow line.

Solvent Used for


Mobile Phase Used Flow Rate Time
Rinse
2-propanol and
Each for 10
New System water, in that 2 mL/min
minutes
order
2-propanol and
Current System Where a Low Polarity Solvent Each for 10
water, in that 2 mL/min
Like Hexane Was Used for the Mobile Phase.
order
minutes 2
Current System Where a Mixture of Water
and Organic Solvent or an Organic Solvent For 10
Water 2 mL/min
Miscible in Water (Methanol, Acetonitrile, minutes
etc.) Was Used for the Mobile Phase.

3 Set the column.


System Guide "3.6.1 Attaching the Column to the Column Oven"

4 Set the method.

Parameter setting values for the test

Item Set Value

Method Pumping Mode ISO


Parameter Mobile Phase Port A
Flow Rate 1 mL/min
Pump
P.Max 40 MPa
Rinse Mode No rinse
Rinsing Volume 100 µL
Rinsing Speed 35 µL/sec
Autosampler 5 µL/sec (When installing Sample Loop (50 µL) )
Sampling Speed
15 µL/sec (When installing the other sample loop)
Cooler Temperature OFF
Injection Volume 5 µL
Column Oven Oven Temperature 40 °C
Wavelength 272 nm
Sampling 100 msec
UV Detector
Response 0.5 sec
Cell Temperature 40 °C

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2 Validation

Item Set Value


Wavelength 272 nm
Sampling 320 msec
PDA Detector Time Constant 640 msec
Cell Temperature 40 °C
Slit 8 nm
WIDTH 2
SLOPE 20000

Data Processing DRIFT 0


Instrument T.DBL 1000
MIN.AREA 20000
STOP.TM 2.5 min

5 Put 1.0 mL of water in a sample vial, add a new septum, and close the cap.
System Guide "3.7 Setting a Sample"

6 Put 1.0 mL of sample in another sample vial, add a new septum, and close the
cap.
System Guide "3.7 Setting a Sample"

7 Start the temperature adjustment of the column oven and begin pumping for
the solvent delivery module.
Check that liquid is flowing from the outlet tubing of the detector and that there is no
fluid leaking from the connection part.

8 Monitor the baseline, and, if a stable baseline is obtained, conduct the zero
adjustment for the detector.

9 Inject 5 µL of water and check whether a peak is observed.

10 Inject 5 µL of sample six times and perform analysis.

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2.5 Other Validations

11 From the six analysis results, calculate the retention time of the peak and the
relative standard deviation (coefficient of variation) of the areas.

: Analysis count

: Retention time (peak area) of each peak

: Mean

: Standard deviation value


2
: Coefficient of variation

"Retention time" and "peak area" formulas

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2 Validation

n Gradient (GRAD.) analysis

This test checks if reproducible data can be obtained from an actual analysis.
This is determined from the coefficients of variation for the peak area and the retention
time of each peak.
As an example, this section explains the gradient analysis of two liquids using mobile
phase port A and mobile phase port B.

Criteria
• "Retention time" coefficient of variation: 0.5 % max.
• "Peak area" coefficient of variation: 1.0 % max.

Parts needed for testing

Part Name Part No. Remarks


Column - Either the column to the left or an
equivalent ODS column can be used.
Shim-Pack XR-ODS S228-41606-92 (Particle diameter: 2.2 µm, Inside diameter:
3.0 mm, Length: 50 mm)
Solvent A (water) : Solvent B (methanol) =
80 : 20(%)
Mobile Phase -
Use HPLC-grade water (distilled water) and
methanol.
20 mg/L caffeine solution (contained in Part
No. S228-45727-91: Caffeine set (5
concentrations)) or one that has been
adjusted using the following procedure.
1 Weigh out 20 mg of caffeine, put it in
a measuring flask with a capacity of
Sample -
100 mL, and dissolve the caffeine in
water so that the total volume becomes
100 mL.
2 Put that 1 mL in a measuring flask with
a capacity of 10 mL, and add water so
that the total volume becomes 10 mL.
Water (HPLC Grade) - -
2-Propanol (HPLC Grade) - -
1.5 mL Sample Vial S228-15652-92 Use 2 sample vials.
1.5 mL Vial Plate
S228-50830-92
or
or Place the 1.5 mL sample vials on the plate.
1.5mL Sample Vial Metal
S228-61615-42
Plate
LabSolutions - Data processing instrument

Before starting the test, rinse the flow line depending on the operating condition
of the instrument.

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2.5 Other Validations

1 Instead of the column, connect the column inlet and outlet tubes to the PEEK
tubing using a coupling.

2 Set the mobile phase and rinse the flow line.

Solvent Used for


Mobile Phase Used Flow Rate Time
Rinse
2-propanol and
Each for 10
2
New System water, in that 2 mL/min
minutes
order
2-propanol and
Current System Where a Low Polarity Solvent Each for 10
water, in that 2 mL/min
Like Hexane Was Used for the Mobile Phase. minutes
order
Current System Where a Mixture of Water
and Organic Solvent or an Organic Solvent For 10
Water 2 mL/min
Miscible in Water (Methanol, Acetonitrile, minutes
etc.) Was Used for the Mobile Phase.

3 Set the column.


System Guide "3.6.1 Attaching the Column to the Column Oven"

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2 Validation

4 Set the method.

Parameter setting values for the test

Item Set Value


Pumping Mode GRAD.
Method
Mobile Phase A 80.0 %
Parameter
Port B 20.0 %
Flow Rate 1 mL/min
Pump P.Max 40 MPa
LPGE Mode Auto
Rinse Mode No rinse
Rinsing Volume 100 µL
Rinsing Speed 35 µL/sec
Autosampler 5 µL/sec (When installing Sample Loop (50 µL) )
Sampling Speed
15 µL/sec (When installing the other sample loop)
Cooler Temperature OFF
Injection Volume 5 µL
Column Oven Oven Temperature 40 °C
Wavelength 272 nm
Sampling 100 msec
UV Detector
Response 0.5 sec
Cell Temperature 40 °C
Wavelength 272 nm
Sampling 320 msec
PDA Detector Time Constant 640 msec
Cell Temperature 40 °C
Slit 8 nm
WIDTH 2
SLOPE 20000

Data Processing DRIFT 0


Instrument T.DBL 1000
MIN.AREA 20000
STOP.TM 2.5 min

5 Put 1.0 mL of water in a sample vial, add a new septum, and close the cap.
System Guide "3.7 Setting a Sample"

6 Put 1.0 mL of sample in another sample vial, add a new septum, and close the
cap.
System Guide "3.7 Setting a Sample"

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2.5 Other Validations

7 Start the temperature adjustment of the column oven and begin pumping for
the solvent delivery module.
Check that liquid is flowing from the outlet tubing of the detector and that there is no
fluid leaking from the connection part.

8 Monitor the baseline, and, if a stable baseline is obtained, conduct the zero
adjustment for the detector.

2
9 Inject 5 µL of water and check whether a peak is observed.

10 Inject 5 µL of sample six times and perform analysis.

11 From the six analysis results, calculate the retention time of the peak and the
relative standard deviation (coefficient of variation) of the areas.

: Analysis count

: Retention time (peak area) of each peak

: Mean

: Standard deviation value

: Coefficient of variation

"Retention time" and "peak area" formulas

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2 Validation

2.5.2 Carryover Test

Using the ratio between the measured peak areas, this validation checks the degree of
retention for the autosampler sample.

CHECK CRITERIA
Carryover £ 0.010 %

Parts needed for testing

Part Name Part No. Remarks


Column - Either the column to the left or an
equivalent ODS column can be used.
Shim-Pack XR-ODS S228-41606-92 (Particle diameter: 2.2 µm, Inside diameter:
3.0 mm, Length: 50 mm)
[For isocratic analysis]
Formulate at the following ratio.
Water : Methanol = 80 : 20(%)
[For gradient (GRAD.) analysis]
Mobile Phase -
Solvent A (water) : Solvent B (methanol) =
80 : 20(%)
Use HPLC-grade water (distilled water) and
methanol.
Sample - 250 mg/L caffeine solution
1.5 mL Sample Vial S228-15652-92 Use 2 sample vials.
1.5 mL Sample Vial Plate
S228-50830-92
or
or Place the 1.5 mL sample vials on this plate.
1.5mL Sample Vial Metal
S228-61615-42
Plate
LabSolutions - Data processing instrument

1 Put 1 mL of sample in a sample vial, add a septum, and close the cap.
System Guide "3.7 Setting a Sample"

2 Put 1 mL of mobile phase (water : methanol = 80 : 20(%)) in another sample


vial, add a septum, and close the cap.
System Guide "3.7 Setting a Sample"

3 Set the sample vial on the plate in the


left rack (L).
• #1: Set the sample vial (sample) prepared
in Procedure 1.
• #2: Set the sample vial (mobile phase)
prepared in Procedure 2.

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2.5 Other Validations

4 Set the column.


System Guide "3.6.1 Attaching the Column to the Column Oven"

5 Set the parameters of the instrument as follows.

Item Set Valu


WIDTH 5
SLOPE 500 2
Data processing instrument
MIN.AREA 10
STOP.TM 5.0 min

• Isocratic analysis
For the parameters other than above, configure the same settings as in Procedure 4 of
"Isocratic analysis" P.84 in "2.5.1 Analysis Reproducibility Test".
"Parameter setting values for the test" P.85

• Gradient (GRAD.) analysis


For the parameters other than above, configure the same settings as in Procedure 4 of
"Gradient (GRAD.) analysis" P.88 in "2.5.1 Analysis Reproducibility Test".
"Parameter setting values for the test" P.90

6 Inject 5 µL of a 250 mg/L caffeine solution sample.

7 Next, inject 5 µL of mobile phase as a blank sample.

8 Calculate the ratio (%) of the peak area value of the sample to the peak area
obtained from the 250 mg/L caffeine solution sample.

Carryover = (Peak area of blank sample / Peak area of caffeine solution sample) x 100
Carryover £ 0.010

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2 Validation

2.5.3 UV Detector Absorbance Linearity Test

This section explains the linearity check of the absorbance value.


The absorbance is measured when the concentration of a caffeine solution varies so as
to check whether the absorbance value of the caffeine solution has linearity with the
caffeine concentration.
There are two checking methods: using an autosampler and using a syringe.

To use an autosampler, "2.5.1 Analysis Reproducibility Test" P.83 should be


conducted and the reproducibility of the injection volume should be checked to
see if it is normal.

n When using an autosampler

CHECK CRITERIA
Absorbance deviation of each concentration: ±5.0 % max.

Parts needed for testing

Part Name Part No. Remarks


1st solvent: 20 mg/L
2nd solvent: 30 mg/L
Caffeine Set
S228-45727-41 3rd solvent: 40 mg/L
(5 Concentrations)
4th solvent: 50 mg/L
5th solvent: 60 mg/L
Water (HPLC Grade) - Used as a mobile phase A.
The pressure when pumping water at 1 mL/min
Resistor Tube PEEK S228-38994
is about 10 MPa.
Male Nut PEEK S228-18565 Used to connect the resistor tube.
Coupling 1.6C S228-16004-13 Used to connect the resistor tube.
1.5 mL Sample Vial S228-15652-92 Use 5 sample vials.
LabSolutions - Data processing instrument

1 Install a resistor tube.


"2.3.1 Processing Automatic Validation" P.23 step7

2 Set the mobile phase.

3 Close the column oven door.


System Guide "3.6.1 Attaching the Column to the Column Oven"

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2.5 Other Validations

4 Set the method.

Parameter setting values for the test

Item Set Value


Pump Flow Rate 0.3 mL/min
Data processing instrument STOP.TM 1.0 min

For the parameters other than above, configure the same settings as in Procedure 4 of 2
"Isocratic analysis" P.84 in "2.5.1 Analysis Reproducibility Test".

5 Pump at a flow rate of 0.3 mL/min.

6 Analyze the 5th solvent to determine the injection volume.

1 Inject 35 µL.

2 If the height of the detection peak does not fall between 1.4 AU and 1.7 AU,
adjust the injection volume.

Changing the injection volume by 10 % causes the peak height to vary by


about 10 %.

7 Analyze the 1st solvent to 5th solvent under the condition determined in
Procedure 6, and read the peak height.

The height can be found more easily if the waveform processing of LabSolutions is used.

LabSolutions Help

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2 Validation

8 Find the absorbance deviation of each concentration and check if those


deviations meet the CHECK CRITERIA.
Calculate the absorbance deviations using the following procedure.

1 Write the sample concentrations [mg/L] in (1) to (5) in the table below.

2 Write the absorbance values (peak heights) [AU] in (A) to (E) in the table below.

3 Calculate the relative sensitivity for each concentration according to the equations
in the table below, and write in 1 to 5.

4 Calculate the standard relative sensitivity using the following equation.


Standard relative sensitivity = (1 + 2 + 3) / 3

5 Calculate the deviation of each concentration using the following equation, and
record.
Deviation[%] = (Relative sensitivity - Standard relative sensitivity) / Standard relative
sensitivity x 100[%]

Sample Absorbance Value Relative


Deviation
Item Concentration (Peak Height) Sensitivity
[%]
[mg/L] [AU] [AU / (mg/L)]
1st
(1) (A) 1 = (A) / (1)
Solvent
2nd
(2) (B) 2 = (B) / (2)
Solvent
3rd
(3) (C) 3 = (C) / (3)
Solvent
4th
(4) (D) 4 = (D) / (4)
Solvent
5th
(5) (E) 5 = (E) / (5)
Solvent

n Causes why the linearity does not meet the CHECK CRITERIA and corrective actions

Cause Corrective Action


Take countermeasures for bubbles.
Air Bubbles Have Entered the Flow
Cell. System Guide "6.1.3 Countermeasures ■
Countermeasures for bubbles"

Rinse the flow cell.


The Flow Cell Is Dirty. Maintenance Guide "5.2 Flow Cell Inspection /
Simple Cleaning"

The Mirror or Aperture Plate Has Conduct linearity recalibration or optical system
Deteriorated. maintenance. (Contact your Shimadzu representative.)

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2.5 Other Validations

2.5.4 PDA Detector Absorbance Linearity Test

This section explains the linearity check of the absorbance value.


The absorbance is measured when the concentration of a caffeine solution varies so as
to check whether the absorbance value of the caffeine solution has linearity with the
caffeine concentration.

When using an autosampler, "2.5.1 Analysis Reproducibility Test" P.83 should be


conducted and the reproducibility of the injection volume should be checked to 2
see if it is normal.

PDA Utility Help "4.5 Linearity Check"

CHECK CRITERIA
Absorbance deviation of each concentration: ±5.0 % max.

Parts needed for testing

Part Name Part No. Remarks


1st solvent: 20 mg/L
2nd solvent: 30 mg/L
Caffeine Set (5 Concentrations) S228-45727-41 3rd solvent: 40 mg/L
4th solvent: 50 mg/L
5th solvent: 60 mg/L
Water (HPLC Grade) - Used as a mobile phase.
The pressure when pumping water at
Resistor Tube PEEK S228-38994
1 mL/min is about 10 MPa.
Male Nut PEEK S228-18565 Used to connect the resistor tube.
Coupling 1.6C S228-16004-13 Used to connect the resistor tube.
1.5 mL Sample Vial S228-15652-92 Use 5 sample vials.
LabSolutions - Data processing instrument

1 Install a resistor tube.


"2.3.1 Processing Automatic Validation" P.23 step7

2 Set the mobile phase.

3 Close the column oven door.


System Guide "3.6.1 Attaching the Column to the Column Oven"

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2 Validation

4 Set the method.

Parameter setting values for the test

Item Set Value


Pump Flow rate 0.3 mL/min
PDA detector Slit 1.2 nm or 8 nm
LabSolutions STOP.TM 1.0 min

For the parameters other than above, configure the same settings as in Procedure 4 of
"Isocratic analysis" P.84 in "2.5.1 Analysis Reproducibility Test".

5 Pump at a flow rate of 0.3 mL/min.

6 Analyze the 5th solvent to determine the injection volume.

1 Inject 35 µL.

2 If the height of the detection peak does not fall between 1.4 AU and 1.7 AU,
adjust the injection volume.

Changing the injection volume by 10 % causes the peak height to vary by


about 10 %.

7 Analyze the 1st solvent to 5th solvent under the condition determined in
Procedure 6, and read the peak height.

The height can be found more easily if the waveform processing of LabSolutions is used.

LabSolutions Help

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2.5 Other Validations

8 Find the absorbance deviation of each concentration and check if those


deviations meet the CHECK CRITERIA.
Calculate the absorbance deviations using the following procedure.

1 Write the sample concentrations [mg/L] in (1) to (5) in the table below.

2 Write the absorbance values (peak heights) [AU] in (A) to (E) in the table below.

3 Calculate the relative sensitivity for each concentration according to the equations 2
in the table below, and write in 1 to 5.

4 Calculate the standard relative sensitivity using the following equation.


Standard relative sensitivity = (1 + 2 + 3) / 3

5 Calculate the deviation of each concentration using the following equation, and
record.
Deviation[%] = (Relative sensitivity - Standard relative sensitivity) / Standard relative
sensitivity x 100[%]

Absorbance
Sample
Value Relative Sensitivity Deviation
Item Concentration
(Peak Height) [AU / (mg/L)] [%]
[mg/L]
[AU]
1st
(1) (A) 1 = (A) / (1)
Solvent
2nd
(2) (B) 2 = (B) / (2)
Solvent
3rd
(3) (C) 3 = (C) / (3)
Solvent
4th
(4) (D) 4 = (D) / (4)
Solvent
5th
(5) (E) 5 = (E) / (5)
Solvent

n Causes why the linearity does not meet the CHECK CRITERIA and corrective actions

Cause Corrective Action


Take countermeasures for bubbles.
Air Bubbles Have Entered the Flow
Cell. System Guide "6.1.3 Countermeasures ■
Countermeasures for bubbles"

Rinse the flow cell.


The Flow Cell Is Dirty. Maintenance Guide "6.2 Flow Cell Inspection /
Simple Cleaning"

The Mirror or Aperture Plate Has Conduct linearity recalibration or optical system
Deteriorated. maintenance. (Contact your Shimadzu representative.)

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2 Validation

2.6 Check the Hardware Validation Results


The automatic validation and performance check results can be checked.
Check from the [Log] tab in the [Maintenance] screen.

The results can also be displayed on the [System Check] screen.

"2.2 System Check" P.5

1 Press (Menu) from the Main


analysis screen.
The [Menu] screen opens.

2 Press [Maintenance].
The [Maintenance] screen is displayed.

3 Press (Log).

The [Log] tab is displayed.

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2.6 Check the Hardware Validation Results

4 Press [Validation Log].

The [Validation Log] screen opens.


For the items and descriptions, see "2.2.3
List of System Check Results" P.12.
System Guide "3.6.1 Attaching the
Column to the Column Oven"

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3 Calibration
This chapter explains "calibration" procedures to adjust the instrument so as to meet the
CHECK CRITERIA.

3.1 Calibration Overview


Calibration is divided into two types: "hardware calibration" for calibrating any selected
item of a unit, and "system adjustment" for adjusting the rack and injection port
positions.

3.2 Hardware Calibration

n Calibration overview

Any individual item of a unit can be calibrated. On the [Hardware Calibration] screen of
the [Maintenance] screen, select an item and run calibration while operating the screen.
A PDA detector system model has some items that can be calibrated by the accessory
PDA Utility. For details, refer to the Help section for the PDA Utility.

Hardware calibration is a calibration task. Basically no fixed CHECK CRITERIA for


calibration exists. Although the CHECK CRITERIA is displayed on the screen, they
are CHECK CRITERIA for testing and displayed as reference, but this does not
mean the hardware calibration has met the CHECK CRITERIA. In order to check
performance after hardware calibration, testing (hardware validation
performance check) is required.

n Calibration items

Item Calibration Items Ref.


Flow rate accuracy calibration P.106
Pump
Gradient concentration accuracy calibration P.109
Injection volume accuracy calibration P.112
Autosampler Sample cooler temperature accuracy calibration (model
P.117
with sample cooler)
Temperature accuracy calibration P.121
Column Oven
Gas sensor operation calibration P.126
Wavelength accuracy calibration P.129
UV Detector Absorbance accuracy calibration (external standard
P.132
filter)
Wavelength accuracy calibration P.137
Absorbance accuracy calibration (external standard
PDA Detector P.140
filter)
Exposure time / W lamp voltage setting P.143

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3.2 Hardware Calibration

Item Calibration Items Ref.


Other Parts Leak sensor calibration P.146

Items that can be calibrated by the PDA Utility (PDA detector system model)
The PDA Utility calibrates the following items. For operation details, refer to the Help
section for the PDA Utility.
• Exposure time setting
• Wavelength calibration

3.2.1 Displaying the Hardware Calibration Items

This section explains how to display each item on the [Hardware Calibration] screen.
3
1 Press (Menu).

The [Menu] screen opens.

2 Press [Maintenance].
The [Maintenance] screen is displayed.

3 Press (Calibration).

The [Login] screen is displayed.

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3 Calibration

4 Enter the System pass ID Number and


press [OK].
The [Calibration] tab is displayed.

5 Press [Hardware Calibration].


The [Hardware Calibration] screen is
displayed.

6 Press the tab of the unit, then select


and press the item to be calibrated.

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7 Follow the on-screen instructions to operate.


Instructions vary depending on the item. The following example gives instructions for
[Flow rate accuracy].

No. Name Explanation


1 Operations Guide Area Displays the operating instructions and buttons.
2 Entire Procedure Displays the entire procedure.
Stop Cancels the operation and closes the screen.
Back Displays the previous screen.
Next Displays the next screen.
Displayed when the results are given after all the
Finish procedures are finished.
Closes the screen.
3 Current Procedure Displays the current procedure position in light gray.

3.2.2 Pump

The following items can be calibrated with the pump.

Item Explanation Ref.


Flow Rate Accuracy Calibrates based on the measurement results of the
P.106
Calibration time required for pumping.
Gradient Concentration Calibrates based on the measurement results of the
P.109
Accuracy Calibration absorbance for the set concentration.

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3 Calibration

n Flow rate accuracy calibration

The mobile phase (water) is pumped at 1 mL/min. Use a stopwatch to record the time
it takes to pump 5 mL. Based on the recorded time, the flow rate accuracy is measured.
After measurement, the flow rate is automatically calibrated based on the measurement
results.

Parts needed for testing

Part Name Part No. Remarks


Water (HPLC Grade) - -
The pressure when pumping water at 1
Resistor Tube PEEK S228-38994
mL/min is about 10 MPa.
Male Nut PEEK S228-18565 Used to connect the resistor tube.
Coupling 1.6C S228-16004-13 Used to connect the resistor tube.
Measuring Cylinder (Volume:
Prepare a validated one. Prepare a
10 mL) or
- measuring flask that comes with
Measuring Flask (Volume: 5
inspection results.
mL)
Stopwatch - Prepare a calibrated stopwatch.
Plumbing Tube (ID 0.3 mm, OD
S228-18495-06 -
1/16'', 1 m)

1 Display the [Hardware Calibration] screen.


"3.2.1 Displaying the Hardware Calibration Items" P.103

2 Press the [Pump] tab and then press [Flow rate accuracy].
The [Flow rate accuracy] screen is displayed.

3 Install a resistor tube.


"2.3.1 Processing Automatic Validation" P.23 step7

4 Set water as the mobile phase.

5 Press [Next].

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6 Select a mobile phase port to be used,


and then press [Next].

When FCV-11AL or FCV-11ALS is


connected
• The flow line set for the
method is used.
• Check for loose fittings.

Operation Guide "5.1.1 Pump"

7 Press [Start].
Purging starts and the screen showing the 3
purge progress is displayed.

Press [Next] to skip purging and start


pumping.

When purging has finished, or after


pressing [Next], pumping starts.

8 Check the chromatogram, and after


pumping has stabilized, press [Next].

Even if [Next] is not pressed, the


next screen is automatically
displayed after about 10
minutes.

9 Insert the end of the tube into the validated measuring cylinder or measuring
flask, and use a stopwatch to measure the time it takes to pump 5 mL.
"Flow rate accuracy test" P.32 step9

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3 Calibration

10 Enter the measured time and then


press [Next].
The measurement results are displayed.

11 Check the measurement results, and to


calibrate based on these results, press
[Calibration].
Calibration starts and the Confirmation
screen opens.

If calibration will not be performed,


press [Finish].

12 Press [Close].
The Confirmation screen closes.

13 Press [Finish].

14 Remove the resistor tube and close the


column oven door.

15 Check the flow rate accuracy.


"Flow rate accuracy test" P.32

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n Gradient concentration accuracy calibration

Use water and caffeine solution to perform binary low-pressure gradient. Absorbance for
each set concentration is measured and the concentration accuracy is calculated.
After measurement, the gradient concentration is automatically calibrated based on the
measurement results.

Parts needed for testing

Part Name Part No. Remarks


Water (HPLC Grade) - -
10 mg/L of Caffeine Solution - -
The pressure when pumping water at 1
Resistor Tube PEEK S228-38994
mL/min is about 10 MPa.
Male Nut PEEK S228-18565 Used to connect the resistor tube. 3
Coupling 1.6C S228-16004-13 Used to connect the resistor tube.

Measurement conditions

Item Requirements
Set Flow Rate 1 mL/min
Preset Wavelength 272 nm
Column Set Temperature 40 °C
A/B, A/C, or A/D 100/0 % for 10 min
A/B, A/C, or A/D 90/10 % for 5 min
Gradient Program A/B, A/C, or A/D 50/50 % for 5 min
A/B, A/C, or A/D 10/90 % for 5 min
A/B, A/C, or A/D 0/100 % for 5 min

Mobile phase ports to be used

Select the desired ports to be used from the combinations A/B, A/C, or A/D. Set
a 10 mg/L caffeine solution for mobile phase port A, and set water for mobile
phase ports B, C, D, and the rinse solution port. (Fill with water those ports that
will not be tested.)

1 Display the [Hardware Calibration] screen.


"3.2.1 Displaying the Hardware Calibration Items" P.103

2 Press the [Pump] tab and then press [GE composition].


The [GE composition] screen is displayed.

3 Install a resistor tube.


"2.3.1 Processing Automatic Validation" P.23 step7

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4 Close the column oven door.


System Guide "3.6.1 Attaching the Column to the Column Oven"

5 Set 10 mg/L of caffeine solution for


mobile phase port A, and set water for
B, C, D, and the rinse solution port.
Then press [Next].

When FCV-11AL or FCV-11ALS is


connected
• The flow line set for the
method is used.
• Check for loose fittings.

Operation Guide "5.1.1 Pump"

6 Press [Start].
Rinse starts.

The screen showing the rinse progress is


displayed.

Press [Skip] to stop the rinse


and move to the rinse for the
next port. Press [Next] to stop
the current rinse, display the
next screen, and start the
system equilibration.

After all ports have been completely


rinsed, the next screen is displayed and the
system equilibration under initial
measurement configuration conditions
starts.

Press [Next] to stop the current


system equilibration and display
the next screen.

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When system equilibration has finished,


the next screen is displayed and
absorbance is measured for each
concentration.
After measurement has finished, the [Next]
button is activated.

7 Press [Next].
The results of system checks are displayed.
3

8 Check the measurement results, and to


calibrate based on these results, press
[Calibration].
Calibration starts and the Confirmation
screen opens.

If calibration will not be performed,


press [Exit].

9 Press [Close].
The Confirmation screen closes.

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10 Press [Exit].

11 Check the gradient concentration


accuracy.
"Gradient concentration accuracy
test" P.38

3.2.3 Autosampler

The following items can be calibrated with the autosampler.

Item Explanation Ref.


Calibrates based on the measurement results of the
Injection Volume Accuracy
difference in mass before and after injection of the P.112
Calibration
sample.
Calibrates based on the difference from the
Cooler Temperature
temperature measured using an external P.117
Accuracy Calibration
thermosensor.

n Injection volume accuracy calibration

Use a 1.5 mL sample vial containing sample (water), and measure the mass before and
after injection of 50 µL. Repeat this operation 10 times. Based on the difference in mass,
the injection volume accuracy is calculated.
After measurement, the injection volume is automatically calibrated based on the
measurement results.

Parts needed for testing

Part Name Part No. Remarks


1.5 mL Sample Vial S228-15652-92 -
Water (HPLC Grade) - -
Balance - Use a validated and calibrated balance.
Sample Rack S228-55735-41 Standard accessory

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Part Name Part No. Remarks


1.5 mL Sample Vial Plate S228-50830-92
Place the 1.5 mL sample vials on the
or or
plate.
1.5mL Sample Vial Metal Plate S228-61615-42

• For models with a sample cooler, to prevent condensation within the


autosampler, turn the temperature control of the sample cooler OFF. For the
setting procedure, refer to "2.1 Analysis Screen" in the Operation Guide.
• Before testing, thoroughly flush the mobile phase port to be used and the
rinse solution with water (HPLC grade).
• When the optional sample loop for 50 µL injections is used, set the sample
aspiration rate to 5 µL/sec or less.

3
1 Display the [Hardware Calibration] screen.
"3.2.1 Displaying the Hardware Calibration Items" P.103

2 Press the [Autosampler] tab and then press [Injection Volume accuracy].
The [Injection Volume accuracy] screen is displayed.

3 Put 1 mL of water into the 1.5 mL


sample vial (A), then measure the mass
using the balance.

4 Enter the measured mass.

Enter the value to the 4th


decimal place.

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5 Set the weighed sample vial


containing water and fitted with a
silicone rubber septum on position 1 of
plate 1 (left side) in rack L.

6 Press [Next].

7 Select a mobile phase port to be used,


and then press [Next].
The screen showing the operation progress
is displayed.

When FCV-11AL or FCV-11ALS is


connected
• The flow line set for the
method is used.
• Check for loose fittings.

Operation Guide "5.1.1 Pump"

When injection is completed, the next


screen is displayed.

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8 Remove the 1.5 mL sample vial (A) and


use the balance to measure the mass.

9 Enter the measured mass and then


press [Next]. 3

Enter the value to the 4th


decimal place.

The results of system checks are displayed.

10 Check the measurement results, and to


calibrate based on these results, press
[Calibration].
Calibration starts and the Confirmation
screen opens.

If calibration will not be performed,


press [Exit].

11 Press [Close].
The Confirmation screen closes.

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12 Press [Exit].

13 Check the injection volume accuracy.


"Injection volume accuracy test" P.44

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n Cooler temperature accuracy calibration

Use an external thermosensor and measure the temperature of the sample cooler. The
temperature accuracy of the sample cooler is calculated based on the measured
temperature. The set value of the sample cooler is "15 °C".
After measurement, the sample cooler temperature is automatically calibrated based on
the measurement results.

Parts needed for testing

Part Name Part No. Remarks


Water (HPLC Grade) - -
External Thermosensor - Use a calibrated sensor.
Sample Rack S228-55735-41 Standard accessory

1.5 mL Sample Vial Metal Plate S228-61615-42


Place the 1.5 mL sample vials on the 3
plate.
1.5 mL Sample Vial S228-15652-92 -

1 Display the [Hardware Calibration] screen.


"3.2.1 Displaying the Hardware Calibration Items" P.103

2 Press the [Autosampler] tab and then press [Cooler Temperature accuracy].
The [Cooler Temperature accuracy] screen is displayed.

3 Fit the external thermosensor to the


sample vial.
1 Put 1 mL of water into a 1.5 mL
sample vial (A), and insert the
external thermosensor (B) through
the septum until it touches the
vial bottom.

2 Set the sample vial with the


external thermosensor attached
into position 27 on the 1.5 mL
sample vial Metal plate.

Check that, when


setting it, the sample
vial is correctly fitted
to the plate.

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3 Calibration

3 Remove the right and left sample


racks, and set the plate fitted
with the external thermosensor
on plate 3, rack R. (Only set
plates on the left front, left back,
and right back sides.)

4 Insert the sample racks.

When inserting the


sample racks, ensure
that the external
thermosensor cable
does not get pinched
inside the
instrument. If the
cable is caught, it
will prevent the
sample racks from
being fully inserted,
resulting in
inaccurate testing.

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4 Press [Next].
The sample cooler starts temperature
control at a set temperature of 25 °C.

When the temperature of the sample


cooler stabilizes, the next screen is
displayed.

If [Next] is pressed, the next 3


screen is displayed even if the
temperature of the sample
cooler is not stable.

5 Read the temperature of the sample vial's external thermosensor.

6 Enter the read temperature and then


press [Next].
The results of system checks are displayed.

7 Check the measurement results, and to


calibrate based on these results, press
[Calibration].
Calibration starts and the Confirmation
screen opens.

If calibration will not be performed,


press [Finish].

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8 Press [Close].
The Confirmation screen closes.

9 Press [Finish].

10 Check the cooler temperature accuracy.


"Sample cooler set temperature
accuracy test" P.47

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3.2.4 Column Oven

The following items can be calibrated with the column oven.

Item Explanation Ref.


Calibrates based on the difference between the
Temperature Accuracy
temperature measured using an external thermosensor P.121
Calibration
and the actual temperature.
Gas Sensor Operation Injects ethanol into the oven and calibrates based on
P.126
Calibration the operating status of the sensor.

n Temperature accuracy calibration

Select [1 Point] when only 1 oven temperature is set. Select [2 Points] when 2 or more
oven temperatures are set. Set any temperature for running calibration by using the 3
following procedure.
Set a calibration temperature. After temperature stabilization, use the external
thermosensor to measure the temperature of the column holder. The oven temperature
can be found from the measured temperature.
After measurement, the oven temperature is automatically calibrated based on the
measurement results.

To calibrate with 2 points, for the first point, set to 60 °C or below for the
calibration temperature, and for the second point, set to at least 20 °C higher.

Parts needed for testing

Part Name Part No. Explanation


External Thermosensor - Use a calibrated sensor.
Use to attach the external thermosensor
Aluminum Tape -
to the column holder.

1 Display the [Hardware Calibration] screen.


"3.2.1 Displaying the Hardware Calibration Items" P.103

2 Press the [Oven] tab and then press [Temperature accuracy].


The [Temperature accuracy] screen is displayed.

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3 Attach the external thermosensor.


1 Open the column oven door.

2 Use 1 cm aluminum tape (B) to fix


the end of the external
thermosensor (A) to the column
holder as in the diagram.

If necessary, tape at
2 or more points to
ensure that the
external
thermosensor will
not fall off the
column holder.

3 Close the column oven door.


System Guide "3.6.1
Attaching the Column to
the Column Oven"

4 Select the number of points to be


measured.

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3.2 Hardware Calibration

5 Press [Next].

6 Enter the calibration temperature.

• When the number of 3


measurement points is 1, only
[First] is displayed.
• To calibrate with 2 points, for
the first point, set to 60 °C or
below for the calibration
temperature, and for the
second point, set to at least
20 °C higher. If the settings
are incorrect, the message
"The entered inspection
temperature is incorrect.
Re-enter the value." is
displayed. Press [Close] and
enter the temperatures again.

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7 Press [Next].

The screen shown on the right is displayed


and temperature control starts.
When the temperature stabilizes, the
screen indicating the next procedure is
automatically displayed.

8 Enter the temperature measured using


the external thermosensor, and then
press [Next].
The results of system checks are displayed.

When calibrating with 2 points,


the temperature control for the
second point starts. The
Operation screens for
procedures 7 and 8 are
displayed again.

9 Check the measurement results, and to


calibrate based on these results, press
[Calibration].
Calibration starts and the Confirmation
screen opens.

If calibration will not be performed,


press [Exit].

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10 Press [Close].
The Confirmation screen closes.

11 Press [Exit].
3
12 Check the temperature accuracy.
"Column oven set temperature
accuracy / Temperature control
accuracy test" P.50

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n Gas sensor operation calibration

The operating status of the gas sensor is checked, and the gas sensor sensitivity is
calibrated.
After measurement, the gas sensor operation is automatically calibrated based on the
measurement results.

Parts needed for testing

Part Name Part No. Remarks


Use one with a volume of 50 µL min.,
Syringe - a needle length of about 50 mm and a
needle outer diameter of 0.5 mm min.
Ethanol - About 50 µL

Measurement conditions

Item Requirements
Oven Temperature 50 °C

If gas sensor calibration is performed with condensation water generated in the


column oven due to temperature cooling control, the gas sensor cannot be
calibrated correctly. Therefore, dry the inside of the oven naturally before
calibration.

1 Display the [Hardware Calibration] screen.


"3.2.1 Displaying the Hardware Calibration Items" P.103

2 Press the [Oven] tab and then press [Gas Sensor].


The [Gas Sensor] screen is displayed.

3 Check the tubing.

1 Open the column oven door and


check for column or tubing
installation problems.

2 Close the column oven door.


System Guide "3.6.1
Attaching the Column to
the Column Oven"

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4 Press [Next].

The screen shown on the right is displayed


and temperature control starts.
When the temperature stabilizes, the
message "Press "Next" button to start
measurement." is displayed. 3
Press [Next].

5 Use a syringe (B) to inject about 50 µL


of ethanol into the interior of the
column oven through the hole for
tubing (A).
When measurement has finished, the
message "Measurement was completed.
Press the "Next" button." is displayed.

Check that the needle of the


syringe is inserted all the way
and then inject ethanol.

6 Press [Next].
The results of system checks are displayed.

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7 Check the measurement results, and to


calibrate based on these results, press
[Calibration].
Calibration starts and the Confirmation
screen opens.

If calibration will not be performed,


press [Exit].

8 Press [Close].
The Confirmation screen closes.

9 Press [Exit].

10 Check the gas sensor operation.


"Gas sensor test" P.54

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3.2 Hardware Calibration

3.2.5 UV Detector

The following items can be calibrated with the UV detector.

Item Explanation Ref.


Calibrates based on the measurement results of the
Wavelength Accuracy
difference between the preset wavelength and the P.129
Calibration
measured wavelength.
Calibrates based on two factors: 1) the absorbance
Absorbance Accuracy
difference between an unmounted and mounted
Calibration P.132
external standard filter, and 2) the absolute
(External Standard Filter)
absorbance of the external standard filter.

n Wavelength accuracy calibration


3
The difference between the preset wavelength and the measured wavelength is
calibrated. The lambda max*1 of the built-in filter (holmium oxide filter) and the
656.1 nm emission line of the D2 lamp (deuterium (D2) lamp) are measured, and the
instrument automatically assesses whether both are within ±1.0 nm.
After measurement, the wavelength accuracy is automatically calibrated based on the
measurement results.
*1 The value varies depending on the instrument (holmium oxide filter). For the lambda max
wavelength, see the criteria (240.4 nm to 241.7 nm) in the system check result screen of Procedure
7.

Parts needed for testing

Part Name Part No. Remarks


Water (HPLC Grade) - -

1 Display the [Hardware Calibration] screen.


"3.2.1 Displaying the Hardware Calibration Items" P.103

2 Press the [Detector] tab and then press [Wavelength accuracy].


The [Wavelength accuracy] screen is displayed.

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3 Select a mobile phase port to be used,


and then press [Next].
The next screen is displayed.

When FCV-11AL or FCV-11ALS is


connected
• The flow line set for the
method is used.
• Check for loose fittings.

Operation Guide "5.1.1 Pump"

4 Press [Start].
Purging starts and the screen showing the
purge progress is displayed.

Press [Next] to skip purging and start


measurement.

When purging has finished, or after


pressing [Next], measurement starts.

5 Wait for measurement to finish.


When measurement has finished, the
message "Measurement was completed." is
displayed.

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6 Press [Next].
The results of system checks are displayed.

7 Check the measurement results, and to


calibrate based on these results, press
[Calibration]. 3
Calibration starts and the Confirmation
screen opens.

If calibration will not be performed,


press [Exit].

8 Press [Close].
The Confirmation screen closes.

9 Press [Exit].

10 Check the wavelength accuracy.


"Wavelength accuracy / Lamp energy
test" P.57

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n Absorbance accuracy calibration (external standard filter)

The absorbance is measured twice, once with the external standard filter not mounted
and once with the filter mounted. Based on the difference and the absolute absorbance
of the external standard filter, the absorbance accuracy of the detector is calculated.
After measurement, the absorbance accuracy is automatically calibrated based on the
measurement results.

Parts needed for testing

Part Name Part Name Remarks


Absorbance Accuracy
S228-57174-41 -
Measurement Cell
Optical Filter to Calibrate
Absorbance - JQA-manufactured JCRM230A
(External Standard Filter)
Water (HPLC Grade) - -

1 Display the [Hardware Calibration] screen.


"3.2.1 Displaying the Hardware Calibration Items" P.103

2 Press the [Detector] tab and then press [Absorbance accuracy].


The [Absorbance accuracy] screen is displayed.

3 Enter the wavelength and actual


absorbance value of the optical filter
to calibrate absorbance (external
standard filter), and then press [Next].
The next screen is displayed.

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4 Attach the absorbance accuracy


measurement cell.
1 Open the controller panel.

2 Remove the flow cell and attach


the absorbance accuracy
measurement cell (A).
3
Do not mount the
optical filter to
calibrate absorbance
(external standard
filter) at this stage in
the procedure.
Mount the filter in
Procedure 6.

3 Close the controller panel.

5 Press [Next].
Absorbance (background absorbance) with
the filter not mounted is adjusted to "0"
and the [Filter Installing] screen is
displayed.

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6 Mount the optical filter to calibrate


absorbance (external standard filter).
1 Open the controller panel.

2 Mount the optical filter to


calibrate absorbance (external
standard filter) (A) to the
absorbance accuracy measurement
cell.

3 Close the controller panel.

7 Press [Start].
With the optical filter to calibrate
absorbance (external standard filter)
mounted, measurement of the absorbance
starts.

When measurement has finished, the


message "Measurement was completed." is
displayed.

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3.2 Hardware Calibration

8 Press [Next].
The results of system checks are displayed.

9 Check the measurement results, and to


calibrate based on these results, press
[Calibration]. 3
Calibration starts and the Confirmation
screen opens.

If calibration will not be performed,


press [Exit].

10 Press [Close].
The Confirmation screen closes.

11 Press [Exit].

12 Check the absorbance accuracy.


"Absorbance accuracy test (external
standard filter)" P.61

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3.2.6 PDA Detector

The following items can be calibrated with the PDA detector.

Item Explanation Ref.


Calibrates based on the measurement results of the
Wavelength Accuracy
difference between the preset wavelength and the P.137
Calibration
measured wavelength.
Calibrates based on two factors: 1) the absorbance
Absorbance Accuracy
difference between an unmounted and mounted
Calibration P.140
external standard filter, and 2) the absolute
(External Standard Filter)
absorbance of the external standard filter.
Exposure Time / W Lamp Sets the optimal exposure time and the W lamp
P.143
Voltage Setting voltage.

The "wavelength accuracy calibration" and "exposure time / W lamp voltage


setting" can be run by the PDA utility.
For operation details, refer to the Help section for the PDA Utility.

"3.1 Exposure Time Setting" and "3.2 Wavelength Calibration" in the PDA Utility Help

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3.2 Hardware Calibration

n Wavelength accuracy calibration

The difference between the preset wavelength and the measured wavelength is
calibrated. Using the emission line of the D2 lamp (deuterium (D2) lamp) and the built-in
filter (holmium oxide filter) lambda max, the wavelength accuracy is checked.
After measurement, the wavelength accuracy is automatically calibrated based on the
measurement results.

Wavelengths checked by testing

Item Wavelength
241.5 nm
Holmium Oxide Filter Lambda Max
360.8 nm
486.0 nm
Emission Lines of the D2 Lamp
656.1 nm*
3
The above wavelengths assume that a 1.2 nm slit is used. When an 8 nm slit is
used, only the wavelength marked with * is checked. (The 1.2 nm and 8 nm slits
are automatically changed.)

Parts needed for testing

Part Name Part No. Remarks


Water (HPLC Grade) - -

1 Display the [Hardware Calibration] screen.


"3.2.1 Displaying the Hardware Calibration Items" P.103

2 Press the [Detector] tab and then press [Wavelength accuracy].


The [Wavelength accuracy] screen is displayed.

3 Select a mobile phase port to be used,


and then press [Next].
The next screen is displayed.

When FCV-11AL or FCV-11ALS is


connected
• The flow line set for the
method is used.
• Check for loose fittings.

Operation Guide "5.1.1 Pump"

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3 Calibration

4 Press [Start].
Purging starts and the screen showing the
purge progress is displayed.

Press [Next] to skip purging and start


measurement.

When purging has finished, or after


pressing [Next], measurement starts.

5 Wait for measurement to finish.


When measurement has finished, the
message "Measurement was completed." is
displayed.

6 Press [Next].
The results of system checks are displayed.

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3.2 Hardware Calibration

7 Check the measurement results, and to


calibrate based on these results, press
[Calibration].
Calibration starts and the Confirmation
screen opens.

If calibration will not be performed,


press [Exit].

8 Press [Close].
The Confirmation screen closes.
3

9 Press [Exit].

10 Check the wavelength accuracy.


"Wavelength accuracy / Lamp energy
test" P.68

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3 Calibration

n Absorbance accuracy calibration (external standard filter)

The absorbance is measured twice, once with the external standard filter not mounted
and once with the filter mounted. Based on the difference and the absolute absorbance
of the external standard filter, the absorbance accuracy of the detector is calculated.
After measurement, the absorbance accuracy is automatically calibrated based on the
measurement results.

Wait for 90 minutes or more after turning on a lamp. Because absorbance is not
stabilized immediately after lamp lighting.

Parts needed for testing

Part Name Part No. Remarks


S228-35011-91 can also be
Absorbance Accuracy Measurement Cell S228-57174-41
used.
Optical Filter to Calibrate Absorbance
- JQA-manufactured JCRM230A
(External Standard Filter)
Water (HPLC Grade) - -

1 Display the [Hardware Calibration] screen.


"3.2.1 Displaying the Hardware Calibration Items" P.103

2 Press the [Detector] tab and then press [Absorbance accuracy].


The [Absorbance accuracy] screen is displayed.

3 Enter the wavelength and fixing


absorbance of the optical filter to
calibrate absorbance (external
standard filter), and then press [Next].
The next screen is displayed.

4 Attach the absorbance accuracy


measurement cell.
1 Open the controller panel.

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3.2 Hardware Calibration

2 Remove the flow cell and attach


the absorbance accuracy
measurement cell (A).

Do not mount the


optical filter to
calibrate absorbance
(external standard
filter) at this stage in
the procedure.
Mount the filter in
Procedure 6.

3 Close the controller panel.

3
5 Press [Next].
Absorbance (background absorbance) with
the filter not mounted is adjusted to "0"
and the [Filter Installing] screen is
displayed.

6 Mount the optical filter to calibrate


absorbance (external standard filter).
1 Open the controller panel.

2 Mount the optical filter to


calibrate absorbance (external
standard filter) (A) to the
absorbance accuracy measurement
cell.

3 Close the controller panel.

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3 Calibration

7 Press [Start].
With the optical filter to calibrate
absorbance (external standard filter)
mounted, measurement of the absorbance
starts.

When measurement has finished, the


message "Measurement was completed." is
displayed.

8 Press [Next].
The results of system checks are displayed.

9 Check the measurement results, and to


calibrate based on these results, press
[Calibration].
Calibration starts and the Confirmation
screen opens.

If calibration will not be performed,


press [Exit].

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3.2 Hardware Calibration

10 Press [Close].
The Confirmation screen closes.

11 Press [Exit].
3
After the check, remove the
optical filter to calibrate
absorbance (external standard
filter) immediately. The filter
may be damaged.

12 Check the absorbance accuracy.


"Absorbance accuracy test (external standard filter)" P.71

n Exposure time / W lamp voltage setting

The exposure time of the D2 lamp is set according to the slit widths (1.2 nm and 8 nm).
When the optional W lamp is connected, the exposure time and voltage are also set.

Parts needed for testing

Part Name Part No. Explanation


Water (HPLC Grade) - -

1 Display the [Hardware Calibration] screen.


"3.2.1 Displaying the Hardware Calibration Items" P.103

2 Press the [Detector] tab and then press [Accumulation time].


The [Accumulation time] screen is displayed.

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3 Calibration

3 Select a mobile phase port to be used,


and then press [Next].

When FCV-11AL or FCV-11ALS is


connected
• The flow line set for the
method is used.
• Check for loose fittings.

Operation Guide "5.1.1 Pump"

4 Press [Start].
Purging starts and the screen showing the
purge progress is displayed.

Press [Next] to skip purging and start


measurement.

When purging has finished, or after


pressing [Next], measurement starts.

5 Wait for measurement to finish.


When measurement has finished, the
message "Measurement was completed." is
displayed.

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3.2 Hardware Calibration

6 Check the measurement results, and to


calibrate based on these results, press
[Calibration].
Calibration starts and the Confirmation
screen opens.

If calibration will not be performed,


press [Exit].

7 Press [Close].
The Confirmation screen closes.
3

8 Press [Exit].

9 Check the lamp energy.


"Wavelength accuracy / Lamp energy
test" P.68

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3 Calibration

3.2.7 Leak Sensor Calibration

When the leak sensor sensitivity is not optimal due to changes in the operating
environment, adjust the leak sensor sensitivity (threshold).
The leak sensor of the detectors, pump, autosampler (HPV: high-pressure valve), and
autosampler (LPV: low-pressure valve) can be adjusted.

Parts needed for calibration

Part Name Part No. Explanation


Water (HPLC Grade) - -
Syringe - -

1 Display the [Hardware Calibration] screen.


"3.2.1 Displaying the Hardware Calibration Items" P.103

2 Press the [Other] tab and then press [Leak sensor calibration].
The [Leak sensor calibration] screen is displayed.

3 Select a leak sensor to be calibrated.

Multiple leak sensors cannot be


selected for one session of
calibration.

4 Pour water under the leak sensor to be calibrated.

UV/PDA detector

1 Open the controller panel.

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3.2 Hardware Calibration

2 Use the syringe to soak the leak


sensor (A) with water.

3 Close the controller panel.

Pump

1 Open the pump door.

2 Use the syringe to soak the leak


sensor (A) with water.

3 Close the pump door.

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Autosampler (HPV)

1 Remove the inserted sample racks


(A) and open the panels and
doors (B) as shown in the
diagram.

2 Remove the 4 knurled screws (A)


and pull the front panel (B)
forward.

3 Use the syringe to soak the leak


sensor (A) with water.

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3.2 Hardware Calibration

4 With the bundle of tubes (C)


passed through the gap (c) on the
left side of the front panel, align
the groove (b) at the bottom of
the front panel with the
protrusion (a) on the main body
frame and insert the front panel.
Attach the 4 knurled screws (A) to
attach the front panel (B).

If the front panel (B)


is not attached
correctly,
condensation may
settle within the
sample cooler. 3

5 Attach the removed sample racks


(A) and close the panels and
doors (B) as shown in the
diagram.

For models with a


sample cooler, do
not conduct testing
without attaching
the front panel.
Condensation will
settle inside the
autosampler. When
running calibration,
turn the sample
cooler temperature
control OFF. (Refer
to "2.1 Analysis
Screen" in the
Operation Guide.)

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3 Calibration

Autosampler (LPV)

1 Open the valve mount door.

2 Use the syringe to soak the leak


sensor (A) with water.

3 Close the valve mount door.

5 Press [Start].
The [Calibration] screen opens and
measurement starts.

The threshold before measurement


continues to be displayed for a short time,
and then a new leak sensor threshold is
calculated and displayed based on the
measured value.

Item Explanation
Dry Value Monitor value while dry
Wet Value Monitor value while wet
Threshold
Threshold before
(Before
measurement
Measurement)
New threshold calculated
Threshold
based on the measured
(New)
value

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3.2 Hardware Calibration

6 Check the measurement results, and to


calibrate based on these results, press
[Calibration].
Calibration starts and the Confirmation
screen opens.

Press [Calibration] first and then


wipe the water off the leak
sensor.
If the water is wiped and
[Calibration] is pressed, the wet
value will change, resulting in
improper calibration.

If calibration will not be performed, press [Close]. 3

7 Wipe the water off the calibrated leak sensor.

8 Press [Close].
The Confirmation screen closes.

9 Press [Close].

10 Check the leak sensor sensitivity.


"Leak sensor test" P.78

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3 Calibration

3.3 System Adjustment


During system adjustment, the rack and injection port positions are adjusted.

3.3.1 Displaying the System Adjustment Items

This section explains how to display each item on the [System Adjustment] screen.

1 Press (Menu).

The [Menu] screen opens.

2 Press [Maintenance].
The [Maintenance] screen is displayed.

3 Press (Calibration).

The [Login] screen is displayed.

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3.3 System Adjustment

4 Enter the System pass ID Number and


press [OK].
The [Calibration] tab is displayed.

5 Press [System Adjustment].


The [System Adjustment] screen is 3
displayed.

6 Press the item to be adjusted.

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3 Calibration

7 Follow the on-screen instructions to operate.


Instructions vary depending on the item. The following example gives instructions for
[Rack Setting].

No. Name Explanation


1 Operations Guide Area Displays the operating instructions and buttons.
2 Entire Procedure Displays the entire procedure.
Stop Cancels the operation and closes the screen.
Back Displays the previous screen.
Next Displays the next screen.
Displayed only when the results are given after the
Finish entire procedure is finished.
Closes the screen.
3 Current Procedure Displays the current procedure position in light gray.

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3.3 System Adjustment

3.3.2 Adjusting the Rack Positions

For each position where a plate is fitted onto a sample rack, adjust the position to which
the needle moves down by following the on-screen instructions.

• Place the plate type selected on the settings screen onto the sample rack. If
a different type is used, adjustment cannot be made properly.
• The stored rack position includes the combinations of the right/left plates, the
rack IDs, and the plate types. Adjust the rack positions for all plates to be set
on each sample rack. When the rack IDs or plate types are changed, adjust
the rack positions again.

System Guide "4.4.1 Setting the Rack ID"

Plate Type
Display Adjustment For 1 mL For 1.5 mL For 4 mL
No. Position 96 Hole 384 Hole
Sample Sample Sample
MTP/DWP MTP/DWP
Vial Vial Vial
Front Left
1 Corner of the 1 1 1 A01 A01
Front Plate
Front Right
2 Corner of the 73 46 22 H01 P01
Front Plate
Rear Left
3 Corner of the 12 9 7 A12 A24
Front Plate
Front Left
4 Corner of the 1 1 1 A01 01
Rear Plate

When adjusting the rack position with sample vial plates, insert the sample vials
into all positions. The rack position cannot be adjusted with only the plates.

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3 Calibration

1 Display the [System Adjustment] screen.


"3.3.1 Displaying the System Adjustment Items" P.152

2 Press [Rack Setting].


The [Rack Setting] screen is displayed.

3 Select a plate type.

1 Press to select a rack

ID.

2 Press [Start teaching].


The [Select Position] screen is
displayed.

4 Remove the front panel.


1 Remove the inserted sample racks
(A) and open the panels and
doors (B) as shown in the
diagram.

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3.3 System Adjustment

2 Remove the 4 knurled screws (A)


and pull the front panel (B)
forward.

3
5 Attach the rack teaching jig to the Z
mount.
1 Align the hole of the rack
teaching jig (B) with the threaded
screw hole (a) at the bottom on
the left side of the Z mount and
attach the cap/knob (A).

6 Select the rack position (L/R) to be


adjusted.

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7 Put the plates on the sample rack and


insert into the instrument.
1 Put the plates (B) on the sample
rack whose position will be set.

Do NOT attach the sample vial cap when adjusting the 4 mL vial plate position.
The cap may make contact with the teaching jig, which can cause errors. If an
error occurs, restart the instrument and readjust the position again.

2 Insert the sample rack (A) on the


side for setting the rack position.

8 Press [Next].
The Adjustment screen for [Position1] is displayed, and the needle moves to [Position1].

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3.3 System Adjustment

9 Finely adjust the needle position.

1 Press [Parallel movement] and


[Vertical movement] to move the
tip of the needle (A) to the center
of the positioning hole (a).

• Use [Vertical
movement] to drop the
tip of the needle to a
height that makes it
easy to check the
relative position
between the needle tip
and the left-right and
front-back positions of 3
the hole.

• Use [Parallel
movement] to align
the tip of the needle
with the center of the
hole.

2 Press [Next].
The next [Position] adjustment
screen is displayed, and the
needle moves near the next
position.

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3 Calibration

Needle distances moved: left-right and front-back, up and down


Pressing [Parallel movement] and [Vertical movement] moves the needle according to the
following table.

Direction Operation

(Left-Right
and
The needle moves in 0.1 mm increments.
Front-Back
Directions)

(Up) The needle moves to the highest point.

First needle drop: 4.0 mm. Second and subsequent drops: 0.2
(Down)
mm.

First needle drop: 4.0 mm. Second and subsequent drops: 1.0
(Down (Large))
mm.

• Press [Stop] to cancel the adjustment during the procedure. The adjustment is
finished without updating the needle position.
• [Vertical movement] is for checking the relative position with the hole by
bringing the tip of the needle closer to a well or a sample vial. This function
is not for deciding the needle stroke.

10 Repeat Procedure 9 to finely adjust the


remaining positions.
After finishing adjustment on the
[Position4] screen and pressing [Finish], the
message "Do you want to save the
teaching information?" is displayed.

Press [Cancel] to close the message


without saving the data.

11 Press [OK].
The message "Remove the teaching jig." is
displayed.

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3.3 System Adjustment

12 Remove the rack teaching jig from the


Z mount.

1 Loosen the cap/knob (A) and


remove the rack teaching jig (B).

After rack position adjustment


is finished, remove the rack
teaching jig and cap/knob.
Failure to do so will result in
interference with the drive
section and will prevent
normal operation. It may also
cause damage to the 3
instrument.

13 Press [Close].

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3 Calibration

14 Attach the front panel.

1 Remove the sample racks.

2 Align the grooves (b) at the


bottom of the front panel with
the protrusions (a) on the main
body frame to insert the front
panel.

3 Place the bundle of tubes (C) in


the gap (c) on the left side of the
front panel.

4 Attach the 4 knurled screws (A).

If the front panel (B)


is not attached
correctly,
condensation may
settle on the rack
plate.

5 Attach the removed sample racks


(A) and close the panels and
doors (B) as shown in the
diagram.

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3.3 System Adjustment

3.3.3 Adjusting the Injection Port Position

Check and adjust the position of the autosampler needle when it moves down to the
injection port of the high-pressure valve. After needle replacement, adjust if the position
has shifted.

Do NOT put hands inside the instrument during autosampler operation.


During adjustment, operation continues even when the front panel is removed.
Never put hands inside the instrument during autosampler operation. There is
a risk of being injured by the needle.

3
1 Display the [System Adjustment] screen.
"3.3.1 Displaying the System Adjustment Items" P.152

2 Press [Injection Port Teaching].


The [Injection Port Teaching] screen is
displayed.

3 Remove the front panel.


"3.3.2 Adjusting the Rack Positions" P.155 Procedure 4

4 Press [Next].
The Adjustment screen for the needle position is displayed.

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3 Calibration

5 Move the tip of the needle to the


center of the upper side of the
injection port.

1 Press [Parallel movement] and


[Vertical movement] to adjust the
left-right and front-back, up and
down positions so that the tip of
the needle (A) drops into the
center of the injection port hole
(B).

To adjust the vertical position, set the tip of the needle at the same
height as the top of the hole (B) on the injection port.

2 Press [Next].
The Confirmation screen is
displayed.

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3.3 System Adjustment

Needle distances moved: left-right and front-back, up and down


Pressing [Parallel movement] and [Vertical movement] moves the needle according to the
following table.

Direction Press and Hold

(Left-Right
and
The needle moves in 0.1 mm increments.
Front-Back
Directions)

(Up) The needle moves to the highest point.

First needle drop: 4.6 mm. Second and subsequent drops: 0.1
(Down)
mm. 3

First needle drop: 4.6 mm. Second and subsequent drops: 1.0
(Down (Large))
mm.

Press [Stop] to cancel the adjustment during the procedure. The adjustment is
finished without updating the needle position.

6 Check that the needle moves down to


the injection port properly.

1 Press [Start].
The sealing operation is
performed twice. Check that the
needle moves down to the
injection port properly.

Press [Back] to readjust


the needle position. The
Adjustment window for
the needle position is
displayed again.

2 After checking, press [Next].


The adjustment position is saved, and then the sealing operation screen is
displayed.

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3 Calibration

7 Press [Start].
The sealing operation is repeated 50 times
to create a smooth seal surface.

8 When the sealing operation has


finished, press [Close].

9 Attach the front panel.


"3.3.2 Adjusting the Rack Positions" P.155

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Index
A R
Absorbance accuracy calibration (external standard resistor tube 27
filter) 132, 140
Absorbance accuracy test 61, 71 S
Adjusting the Injection Port Position 163 Sample cooler set temperature accuracy test 47
Adjusting the Rack Positions 155 Solvent delivery stability test 35
Automatic Validation 23 System Check 5

B T
Balance 44 Temperature accuracy calibration 121
Baseline noise/drift test 64, 74
U
C UV Detector 94
Calibration 102
Column oven set temperature accuracy / V
Temperature control accuracy test 50 Validation 2
Cooler temperature accuracy calibration 117
W
E Wavelength accuracy / Lamp energy test 57, 68
Exposure time / W lamp voltage setting 143 Wavelength accuracy calibration 129, 137

F
Flow rate accuracy calibration 106
Flow rate accuracy test 32

G
Gas sensor test 54
Gas sensor operation calibration 126
Gradient concentration accuracy calibration 109
Gradient concentration accuracy test 38

H
Hardware Calibration 102
holmium oxide filter 57

I
Injection volume accuracy calibration 112
Injection volume accuracy test 44
Items that can be calibrated by the PDA Utility 103

L
Leak Sensor Calibration 146
Leak sensor test 78

O
Other Validations 83

P
PDA Detector 97
Performance Check 30
Pressure limiter test 41

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