Waters ACQUITY UPLC I-Class Parts list manual

ACQUITY UPLC I-Class
System Guide
715003736 / Revision B
Copyright © Waters Corporation 2014
All rights reserved

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Copyright notice
© 2014 WATERS CORPORATION. PRINTED IN THE UNITED STATES OF
AMERICA AND IN IRELAND. ALL RIGHTS RESERVED. THIS
DOCUMENT OR PARTS THEREOF MAY NOT BE REPRODUCED IN ANY
FORM WITHOUT THE WRITTEN PERMISSION OF THE PUBLISHER.
The information in this document is subject to change without notice and
should not be construed as a commitment by Waters Corporation. Waters
Corporation assumes no responsibility for any errors that may appear in this
document. This document is believed to be complete and accurate at the time
of publication. In no event shall Waters Corporation be liable for incidental or
consequential damages in connection with, or arising from, its use. For the
most recent revision of this document, consult the Waters Web site
(waters.com).
Trademarks
ACQUITY, ACQUITY UPLC, Connections INSIGHT, Millennium, PIC,
UPLC, and Waters are registered trademarks, and Auto•Blend Plus, eCord,
Empower, LAC/E, MassLynx, XBridge, and “THE SCIENCE OF WHAT’S
POSSIBLE.” are trademarks of Waters Corporation.
Analyst is a registered trademark of Applied Biosystems/MDS SCIEX.
Keps is a registered trademark of the Shakeproof division of Illinois Tool
Works, Inc.
Phillips is a registered trademark of Phillips Screw Company.
Teflon is a registered trademark of E. I. du Pont de Nemours and Company.
TORX is a registered trademark of Textron Corporation.
PEEK is a trademark of Victrex Corporation.
PEEKsil is a trademark of SGE Analytical Science Pty Ltd.
PharMed and Tygon are registered trademarks of Saint-Gobain Ceramics &
Plastics, Inc.
TORX is a registered trademark of Textron Corporation.
TRITON is a trademark of Union Carbide Corporation.
TWEEN is a trademark of ICI Americas, Inc.

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Other registered trademarks or trademarks are the sole property of their
owners.
Customer comments
Waters’ Technical Communications department invites you to tell us of any
errors you encounter in this document or to suggest ideas for otherwise
improving it. Please help us better understand what you expect from our
documentation so that we can continuously improve its accuracy and
usability.
We seriously consider every customer comment we receive. You can reach us
at [email protected].
Contacting Waters
Contact Waters®with enhancement requests or technical questions regarding
the use, transportation, removal, or disposal of any Waters product. You can
reach us via the Internet, telephone, or conventional mail.
Waters contact information:
Contacting medium Information
Internet The Waters Web site includes contact
information for Waters locations worldwide.
Visit www.waters.com.
Telephone and fax From the USA or Canada, phone 800
252-HPLC, or fax [phone].
For other locations worldwide, phone and fax
numbers appear in the Waters Web site.
Conventional mail Waters Corporation
34 Maple Street
Milford, MA 01757
USA

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Safety considerations
Some reagents and samples used with Waters instruments and devices can
pose chemical, biological, or radiological hazards (or any combination thereof).
You must know the potentially hazardous effects of all substances you work
with. Always follow Good Laboratory Practice, and consult your organization’s
safety representative for guidance.
Considerations specific to the ACQUITY UPLC I-Class
instruments
See also: For safety considerations regarding specific system modules,
consult the appropriate information on the user documentation CD.
High voltage hazard
Safety advisories
Consult Appendix A for a comprehensive list of warning and caution
advisories.
Operating the ACQUITY UPLC I-Class system
When operating the ACQUITY UPLC I-Class system, follow standard
quality-control (QC) procedures and the guidelines presented in this section.
Warning: To avoid electric shock, do not remove protective panels from
the ACQUITY UPLC®I-Class system modules. The components within
are not user-serviceable.

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Applicable symbols
Audience and purpose
This guide is intended for personnel who install, operate, and maintain
ACQUITY UPLC I-Class system modules. It gives an overview of the system’s
technology and operation.
Intended use of the ACQUITY UPLC I-Class system
Waters designed the ACQUITY UPLC I-Class system to perform
chromatographic separations. The system is for research use only and is not
intended for use in diagnostic applications.
Calibrating
To calibrate LC systems, follow acceptable calibration methods using at least
five standards to generate a standard curve. The concentration range for
standards should include the entire range of QC samples, typical specimens,
and atypical specimens.
Symbol Definition
Manufacturer
Authorized representative of the European
Community
Confirms that a manufactured product complies
with all applicable European Community
directives
Australia C-Tick EMC compliant
Confirms that a manufactured product complies
with all applicable United States and Canadian
safety requirements
Consult instructions for use

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When calibrating mass spectrometers, consult the calibration section of the
operator’s guide for the instrument you are calibrating. In cases where an
overview and maintenance guide, not an operator’s guide, accompanies the
instrument, consult the instrument’s online Help system for calibration
instructions.
Quality-control
Routinely run three QC samples that represent subnormal, normal, and
above-normal levels of a compound. Ensure that QC sample results fall within
an acceptable range, and evaluate precision from day to day and run to run.
Data collected when QC samples are out of range might not be valid. Do not
report these data until you are certain that the instrument performs
satisfactorily.
ISM classification
ISM Classification: ISM Group 1 Class B
This classification has been assigned in accordance with IEC CISPR 11
Industrial Scientific and Medical (ISM) instruments requirements. Group 1
products apply to intentionally generated and/or used conductively coupled
radio-frequency energy that is necessary for the internal functioning of the
equipment. Class B products are suitable for use in both commercial and
residential locations and can be directly connected to a low voltage,
power-supply network.

vii
EC authorized representative
Waters Corporation
Stamford Avenue
Altrincham Road
Wilmslow SK9 4AX
United Kingdom
Telephone: [phone]
Fax: [phone]
Contact: Quality manager

viii

Table of Contents ix
Copyright notice ................................................................................................... ii
Trademarks ............................................................................................................ ii
Customer comments ............................................................................................ iii
Contacting Waters ............................................................................................... iii
Safety considerations .......................................................................................... iv
Considerations specific to the ACQUITY UPLC I-Class instruments............. iv
Safety advisories................................................................................................. iv
Operating the ACQUITY UPLC I-Class system ............................................. iv
Applicable symbols .............................................................................................. v
Audience and purpose.......................................................................................... v
Intended use of the ACQUITY UPLC I-Class system ....................................... v
Calibrating ........................................................................................................... v
Quality-control .................................................................................................... vi
ISM classification ................................................................................................. vi
ISM Classification: ISM Group 1 Class B ......................................................... vi
EC authorized representative .......................................................................... vii
1 ACQUITY UPLC I-Class System ......................................................... 1-1
UltraPerformance Liquid Chromatography .............................................. 1-2
Features of the ACQUITY UPLC I-Class system ........................................ 1-3
Binary solvent manager .................................................................................. 1-4
Column compartments .................................................................................... 1-4
Injector mechanisms........................................................................................ 1-4
Detectors........................................................................................................... 1-5
Mass spectrometry........................................................................................... 1-5
System modules ................................................................................................. 1-6
Binary solvent manager .................................................................................. 1-7
Sample manager-FTN ..................................................................................... 1-7
Table of Contents

x Table of Contents
Sample manager-FL ........................................................................................ 1-7
Column heater ................................................................................................. 1-8
30-cm Column heater (optional)...................................................................... 1-8
Column manager (optional)............................................................................. 1-9
Sample organizer (optional) ............................................................................ 1-9
Column technology .......................................................................................... 1-9
Detectors......................................................................................................... 1-10
Local Console Controller (optional)............................................................... 1-11
For additional information ............................................................................ 1-11
2 Optimizing Performance ...................................................................... 2-1
General guidelines ............................................................................................ 2-2
Dispersion ........................................................................................................... 2-3
Carryover ............................................................................................................ 2-4
Contamination ................................................................................................... 2-5
Reproducibility ................................................................................................. 2-5
Cycle time (between injections) .................................................................... 2-5
Preventing leaks ............................................................................................... 2-5
Sample preparation .......................................................................................... 2-6
Particulates ...................................................................................................... 2-6
Matching sample diluents ............................................................................... 2-6
3 Preparing the System ........................................................................... 3-1
Preparing system hardware ........................................................................... 3-2
Powering-on the system .................................................................................. 3-2
Monitoring system module LEDs ................................................................... 3-2
Power LED ....................................................................................................... 3-3
Status LEDs..................................................................................................... 3-3
Enabling the leak sensors ............................................................................... 3-4
Starting up the system .................................................................................... 3-5

Table of Contents xi
Configuring chromatography data software ............................................. 3-5
ACQUITY control panels ................................................................................. 3-6
Binary solvent manager control panel............................................................ 3-6
Sample manager control panel ....................................................................... 3-8
TUV detector control panel ........................................................................... 3-10
Column manager control panel..................................................................... 3-11
Starting the ACQUITY UPLC Console ....................................................... 3-13
A Safety Advisories .................................................................................. A-1
Warning symbols ............................................................................................... A-2
Task-specific hazard warnings........................................................................ A-2
Specific warnings ............................................................................................. A-3
Caution advisory ............................................................................................... A-5
Warnings that apply to all Waters instruments ......................................... A-5
Electrical and handling symbols ................................................................. A-11
Electrical symbols .......................................................................................... A-11
Handling symbols .......................................................................................... A-12
B External Connections .......................................................................... B-1
System tubing connections ............................................................................ B-1
External wiring connections ......................................................................... B-4
ACQUITY UPLC I-Class instrument external wiring connections .............. B-4
Ethernet connections ....................................................................................... B-5
Column heater connection............................................................................... B-5
Signal connections ........................................................................................... B-5
Making signal connections .............................................................................. B-5
Connecting to the electricity source ......................................................... B-11

xii Table of Contents

1-1
1ACQUITY UPLC I-Class System
Waters designed the ACQUITY UPLC®I-Class system for optimum
performance when running difficult assays. Configure it with a mass
spectrometer to take full advantage of the system’s design
enhancements.
Contents:
Topic Page
UltraPerformance Liquid Chromatography 1-2
Features of the ACQUITY UPLC I-Class system 1-3
System modules 1-6

1-2 ACQUITY UPLC I-Class System
UltraPerformance Liquid Chromatography
In 2004, Waters made significant advances in instrumentation and column
design when it introduced UPLC®technology to the field of separation science.
By employing this technology, Waters’ ACQUITY UPLC systems achieve a
marked increase in resolution, speed, and sensitivity in liquid
chromatography, when compared to conventional systems.
UltraPerformance liquid chromatography uses columns packed with round
particles, 1.7 μm-diameter; its operating pressures are as high as 18,000 psi.
The van Deemter equation, an empirical formula that describes the
relationship between linear velocity and column efficiency, considers particle
size as one of the variables. Thus the equation can be used to characterize
theoretical performance across a range of particle sizes.
History of particle size in liquid chromatography:
It is apparent from the figure, above, that using 1.7-μm particles achieves
higher efficiency that persists as the flow rate increases (lower HETP
indicates higher efficiency, HETP = Height Equivalent to a Theoretical Plate).
When operating in this area of the plot, the peak capacity and the speed of a
separation can set limits well beyond those of conventional HPLC technology.

Features of the ACQUITY UPLC I-Class system 1-3
Waters has defined this new level of performance as UltraPerformance liquid
chromatography.
Comparison of chromatographic separations using 5.0-μm and 1.7-μm
particles:
Each separation was performed on a 2.1 × 50 mm column. Chromatographic
conditions for the separations were identical, except for the flow rate, which
was scaled according to particle size.
Features of the ACQUITY UPLC I-Class system
The ACQUTY UPLC I-Class system significantly improves the results of
traditionally challenging separations.
When compared to systems typically used for difficult assays, the I-Class
system confers these advantages:
• Significantly decreased system dispersion
• Rapid injection cycles and sample throughput

1-4 ACQUITY UPLC I-Class System
• Greatly reduced carryover
These advantages provide the following benefits over traditional systems:
• Superior peak capacity
• Higher sensitivity
• Faster data acquisition
• Consistent, reproducible results
The I-Class system is an effective tool for analyzing complex or dilute samples,
providing high resolution and reproducible results for chromatographic
separations.
Binary solvent manager
The ACQUITY UPLC I-Class system employs a binary solvent manager that,
like the system, is optimized for sub-2-μm particle liquid chromatography and
uses reduced fluid volumes. New pressure management capabilities extend
the flow rate envelope for the system to support ballistic gradients. This
support enables you to run higher flow rates and faster cycle times to
accelerate your chromatography, while maintaining the integrity of the
separation.
Column compartments
Column compartments configured with I-Class systems ensure reliable and
robust separations. This is especially important when you transfer methods to
other systems. UPLC applications can benefit from pre-column, mobile-phase
heating to improve chromatographic separations. The ACQUITY UPLC
I-Class column compartments use an active preheater to condition solvent as
it enters the column. The preheater is a heat source that raises the
temperature of the incoming mobile phase and injected sample to the same set
point as that of the column compartment. Active solvent preheaters provide
precise thermal performance with low dispersion for exact control of
chromatographic conditions that remain consistent between systems.
Injector mechanisms
ACQUITY UPLC I-Class system configurations includes a sample manager
with either a fixed-loop or flow-through-needle injector mechanism. This
flexibility allows you to choose the optimum injector mechanism for your

Features of the ACQUITY UPLC I-Class system 1-5
application. Both mechanisms provide high precision with low dispersion and
efficient sample recovery.
The low carryover performance of the injector mechanisms, which enables you
to use a wider sample concentration range, is also beneficial for mass
spectrometry applications.
Detectors
The PDA and TUV detectors available with the I-Class system feature
low-dispersion UV flow-cell technology, which enhances peak resolution.
Mass spectrometry
When you configure an I-Class system with a mass spectrometer, the system’s
design enhancements enable you to separate more peaks with better peak
shapes to aid quantitation. You also benefit from improvements in sensitivity
and flexibility.
See also: Waters.com for information on compatible Waters mass
spectrometers.

1-6 ACQUITY UPLC I-Class System
System modules
See also: Configuring ACQUITY UPLC I-Class System Components (P/N
715003144)
The ACQUITY UPLC I-Class core system includes a binary solvent manager,
a sample manager (flow-through-needle or fixed loop), a column heater
(CH-A), detectors (tunable ultraviolet, photodiode array, eλphotodiode array,
or mass spectrometry), and an ACQUITY UPLC column.
Waters Empower™ chromatography data software, MassLynx™ mass
spectrometry software, standalone ACQUITY UPLC Console software, or
Analyst®software controls the system.
Bottle tray
Detector
Column heater
Sample manager -
FTN or Sample
manager-FL
Binary solvent
manager

System modules 1-7
Binary solvent manager
The Waters ACQUITY UPLC I-Class Binary Solvent Manager delivers
solvent compositions for isocratic and binary gradient methods at flow rates of
0.01 to 2.0 mL per minute. Its features include in-line filters upstream of a
primary check valve, the Waters®Intelligent Intake Valve (i2Valve),
automated priming functions, and daily system-setup routines.
The design of the binary solvent manager (and, indeed, the system overall) is
optimized for sub-2-µm particle liquid chromatography. The upper pressure
limit is set at 124,106 kPa (1241 bar, 18,000 psi) per pump. A maximum flow
rate of 1 mL/min is permitted at system backpressures to 124,106 kPa
(1241 bar, 18,000 psi), which falls within the range of optimal linear velocity
for sub-2-µm particle columns of ID 1.0 mm to 3.0 mm.
See also: The ACQUITY UPLC Binary Solvent Manager Operator's Overview
and Maintenance Information for more details.
Sample manager-FTN
The ACQUITY UPLC I-Class Sample Manager - FTN uses a
flow-through-needle mechanism that differs from a loop-based injector. The
mechanism aspirates a sample from plates and vials and holds it in the
sample needle in preparation for injecting the sample onto the column. The
needle serves as part of the injection flow path when the sample is pushed
onto the column.
Optional extension loops (installed between the sample needle and the
injection valve) can increase the injection volume beyond that of the sample
needle. The sample manager-flow through needle can also dilute samples
using the auto-dilution option
Using the flow-through-needle mechanism, the system operates similarly to
most traditional HPLC systems, which facilitates the transfer of HPLC
methods. The flow-through-needle mechanism also does not require you to
learn new injection modes. Moreover, it achieves injection accuracy and
decreases cycle time for small volume injections. Gradients pass through the
needle during injection, ensuring complete sample recovery.
Sample manager-FL
The sample manager - FL uses a loop-based mechanism to inject samples.
After a puncture needle pierces the well or vial covering, a sample needle

1-8 ACQUITY UPLC I-Class System
emerges from within the puncture needle, to aspirate the sample. In the
partial loop needle overfill injection mode (default), the sample needle is then
removed from the vial and the sample syringe continues to pull the sample
aliquot through the sample needle and through the injection valve until any
pre-sample and the sample injection volume pass through the injection valve.
The valve actuates, switching the sample loop to the load position. The sample
is pushed back toward the needle and the sample volume is then pushed into
the sample loop. The sample loop is moved to the injection position and flow
from the pump pushes the sample onto the column.
See also: ACQUITY UPLC Sample Manager - Fixed Loop Operator’s Overview
and Maintenance Information for information on other injection modes.
Column heater
Column temperature variations can shift peak retention times and alter peak
shapes, increasing the difficulty of achieving precise results. The ACQUITY
UPLC I-Class system’s column heater helps to ensure precise, reproducible
separations by controlling the column temperature.
The column heater (CH-A) heats the column compartment to any temperature
from 20 ºC to 90 ºC, that is at least 5 °C above the ambient temperature. A low
dispersion, active preheating device heats the incoming solvent before it
enters the column. The column compartment can accommodate columns of up
to 4.6 mm I.D. and up to 150 mm length.
30-cm Column heater (optional)
The optional 30-cm column heater (CH-30A) heats the column compartment to
any temperature, from 20 ºC to 90 ºC, that is at least 5 ºC above the ambient
temperature. The 30-cm column heater mounts on the side of the system
stack.
The 30-cm column heater can accommodate UPLC®columns of 4.6 mm, or
less, ID and 150 mm, or less, length and HPLC columns of 4.6 mm, or less, ID
and 300 mm, or less, length. For columns that exceed 4.6 mm ID, use the
ACQUITY UPLC 30-cm column heater/cooler with the appropriate
compatibility kit.
Note: The ACQUITY UPLC 30-cm column heater/cooler is compatible with
I-Class systems. See ACQUITY UPLC Column Compartments Operator’s
Overview and Maintenance Information for details.