Alliance HPLC Systems
The industry-standard HPLC platform with versatile, dependable performance
to support the analytical needs of your laboratory
• Flexible system configurations address the needs and tasks of multiple applications
•Consistent performance that delivers timely, predictable results
•Scalable LC separation chemistries that do not compromise results quality
•Maximum system dependability with minimized downtime
The Waters Alliance HPLC System: Designed for versatility
Inspired by input from our increasingly time-constrained users, industry-benchmark Waters Alliance®
HPLC Systems have been continually expanded and perfected over the years to accommodate
an enormous range of real-world, problem-solving HPLC separation challenges. Synonymous with
dependable performance, highest quality and flexibility of operation, tens of thousands of these
systems are in service worldwide today.
Alliance Systems are able to meet the rigorous requirements of routine analyses in addition to the
performance standards of new-product research and method development. Whether your needs are
in Pharmaceutical, Chemistry, Food Safety, Environmental or Quality Control arenas, Alliance HPLC
Systems offer you a field-proven solution for all of your HPLC requirements—today and in the future.
The flexible and
reliable workhorse
for all of your
HPLC requirements.
Faster HPLC with the Alliance System
COMPOUNDS
USP TAILING
1. 8-Bromoguanosine
1.22
2. Acetanilide
1.15
3. Triamcinolone
1.25
4. Hydrocortisone
1.14
5. 2-amino-7-chloro-5-oxo1.06
5H-[1]benzopyrano[2,3-b]pyridine3-carbonitrile
6. 6a-methyl-17a-hydroxy progesterone 1.17
7. 3-Aminofluoranthene
1.17
8. 2-Bromofluorene
1.12
9. Perylene
1.29
10. Naphthol[2,3-a]pyrene
1.15
CONDITIONS
Column Name:
SunFire™ C18 - 4.6 x 20 mm
IS™, 3.5 μm
Mobile Phase A: 0.1% HCOOH in Water
Mobile Phase B: 0.1% HCOOH in ACN
Flow Rate:
3.0 mL/min
Gradient:
Time
Profile
(min) %A
%B
0.0
100
0
4.0
0
100
Simple start-up, robust data management—
The Allliance HPLC System features a large and intuitive
7
1
0.15
Maximum PSI:
1190
VO=0.26
PEAK CAPACITY: 80
2
0.20
AU
0.10
3
4
8
6
9
LCD-based user interface. Analysts can take advantage
10
5
0.05
of rapid system set-up through AutoStartPLUS routines that
0.00
0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60 3.80
streamline daily startups.
4.00
Minutes
Alliance Systems also collect, analyze, report, and store
Alliance HPLC Systems provide faster run times, without changing
hardware, with Intelligent
Speed™
(IS™)
data through Empower™ Software for LC and LC/MS,
HPLC column chemistries.
or MassLynx™ Software for LC/MS
This example shows a SunFire™ C18 2 cm IS column delivering a
and LC/MS/MS analyses.
gradient separation of 10 diverse compounds in less than four minutes.
Alliance
Other-brand HPLC
2.384
0.0066
1.967
Optimizing Methods with the Alliance HPLC System
0.060
0.0044
AU
AU
0.048
0.0022
0.036
0.0000
0.024
-0.0022
0.012
-0.0044
0.00
0.000
3.00
6.00
9.00
12.00
15.00
18.00
21.00
24.00
27.00
30.00
0.30
0.60
Minutes
0.90
1.20
1.50
1.80
2.10
Minutes
Reduce time to market with the Alliance System by running your methods faster and using more reliable separations.
By taking full advantage of new HPLC chemistries, you can update and optimize your current USP methods.
The Waters Automated Method Development System can facilitate this process for you.
2.40
2.70
3.00
Minimize risks—
Fully integrated fluid leak pathways provide peace of
mind by ensuring that accidental leaks and spills are
safely managed and directed out of harm’s way.
Ensure accuracy of results—
The duration of the needle wash is user definable—
it helps you to intelligently manage sample carryover
concerns without using wash or rinse vials.
Manage the separation process—
Single or multiple columns may
be temperature controlled via a
column heater or column heater/
cooler. Both provide extremely
stable column environments, thanks
to built-in forced air recirculation.
Available integrated switching
valves permit rapid column
selection, enabling unattended
HPLC method development.
Integrated fluidics—
The integrated solvent and sample
management functions ensure
highly consistent system-to-system
performance, bringing the
highest reproducibility to your
laboratory processes.
Manage your sample workload—
Up to 120 industry-standard vials can be
accommodated in five individual sample
Manage your LC solvents—
carousels. Sample queues are quickly
The solvent management system
set up—whether it’s just one sample or
degasses and blends up to four
a sequence of multiple methods from
chromatographic solvents in
different analysts. The sample environment
precise proportions for smooth,
may be temperature-controlled and is
pulse-free solvent delivery, while
compatible with light-sensitive samples.
the latest PerformancePLUS™
check-valve technology assures
Tool-free maintenance—
The Alliance System’s design helps maximize uptime by providing
easy, tool-free access to pistons, seals, and seal-wash seals, as well
as the lower needle-wash seal of the sample manager.
minimal downtime.
Empower Chromatography Data Software
Empower Software is Waters’ market-leading chromatography data software package for advanced data acquisition, management,
processing, reporting, and distribution with Alliance HPLC Systems. From the occasional user to the daily power user, Empower
Software will increase your productivity in running separations and analyzing results.
Empower Software simplifies how you collect and report chromatography test results by letting you choose the user interface
that matches the way you work. Whether you work at a pharmaceutical company concerned about data security, compliance
and validation, or you have an industrial application where ease-of-use and high system availability are top priorities, Empower
Software can help your lab perform more efficiently.
Empower includes capabilities such as automated archiving, automated method development, customizable data reports, integrated
custom calculations, advanced data management (including chemical structures), and an intuitive online answer wizard to help
maximize your lab’s efficiency.
Empower Software’s intuitive interface options help everyone in your lab work better
QuickStart—A single-window access point for data
collection, processing, and reporting
Open Access—Gives the walk-up user, who may not be
a trained chromatographer, the ability to simply select a
method and number of samples, and start a run with the
click of a mouse
Pro—For system administrators or power users who
need total control and compartmental access to all
Empower Software functions
Web—Provides secure, remote access to results through
a standard intranet or Internet connection
Consistent, quality data with ConnectionsAQT
ConnectionsAQT is a patented technology that enhances the qualification
testing and data management of the Alliance HPLC System, its software,
and additional devices.
Using the advanced systems control, data acquisition, and custom
reporting features of Empower Software, ConnectionsAQT provides
documentation to meet GxP and 21 CFR Part 11 requirements.
Qualification Services can be delivered by trained and certified Waters
Compliance Specialists, or Waters can provide you with the workbooks
and training necessary to perform your own system and software
qualifications.
MassLynx Mass Spectrometry Data Software
MassLynx Software is Waters’ high-performance mass spectrometry data
software that provides scientists with the ideal data platform for acquiring,
analyzing, managing, and distributing mass spectrometry information in
conjunction with the Alliance HPLC System.
Built with a sample-centric approach, MassLynx Software offers flexible control
over complex instrument configurations, from single quadrupole
to hybrid quadrupole time-of-flight mass spectrometers.
With its intelligent instrument control and ability to acquire nominal mass,
exact mass, MS/MS and exact mass MS/MS data, MassLynx Software
provides the versatility required in today’s laboratories.
MassLynx offers sophisticated application managers that are tailored
for specific MS analysis, including protein identification, automated
quantification, tracking fractions in purification, and metabonomics.
MassLynx Application Managers, for application-specific
data acquisition and processing
MassLynx Software’s OpenLynx browser summarizes
results in a color-coded map for easy visualization.
Chemists can review the data supporting any
found/not found/tentative assignment by clicking
on the sample location of interest.
ChromaLynx™—For the rapid detection, identification, and semi-quantitative determination of components in complex mixtures
FractionLynx™—Fraction tracking for compound isolation and purification
MarkerLynx™—Comprehensive data processing for metabonomics and biomarker applications, including multivariate statistical analysis
MetaboLynx™—Automates metabolite identification
OpenLynx™—Provides walk-up LC/UV, LC/MS and LC/MS/MS capabilities, automated batch
processing, and improves your ability to share and search results
ProfileLynx™—Automates data processing for chromatography-based physicochemical
property analysis in drug discovery and lead optimization
ProteinLynx™—For relative protein quantification, identification, and characterization
QuanLynx™—For high performance, high throughput quantification, automated MS
and MS/MS method development with QuanOptimize™; includes tools
for 21 CFR Part 11 compliance
TargetLynx™—For advanced quantification through a full range of automatic quality control checks
Advanced data management solutions with Waters Laboratory Informatics
For a comprehensive information management solution for your laboratory, combine Empower or MassLynx software-driven Alliance
HPLC Systems with our informatics software.
The Waters Laboratory Informatics Suite—including eLab Notebook™ and NuGenesis® SDMS (Scientific Data Management
System)—brings together the information management solutions you need to compile, search, and share scientific data, and, at the
end of the day, make the most knowledgeable decisions possible. These tightly integrated software solutions improve how you use
the data generated by your Alliance System for immediate processing, and store and protect information to support future product
development—safeguarding intellectual property.
Alliance offers a wide spectrum of HPLC
100 fg/µl - 1 pg on-column
Eq_STD1pg_A7_002
100
7: MRM of 1 Channel ES+
17.23
TIC
962
Imazalil7.46e3
Area
%
Liquid Chromatography
4
Eq_STD1pg_A7_002
Alliance 2695 HPLC System
Mobile phase A: MeOH/H2O (1:4) + 5mM CH3CO2NH4
Mobile phase B: MeOH/H2O (9:1) + 5mM CH3CO2NH4
®
Column: Waters Atlantis
%
0
Eq_STD1pg_A7_002
Flow: 220 uL/min
8.94
4511
100
C18 2.1 mm id, 100 mm with 3
µm particle size, 30 ˚C
14.71
3041
100
%
1
Eq_STD1pg_A7_002
7.19
538
100
Gradient from 100% A to 100% B in 15 min
%
2
Eq_STD1pg_A7_002
6.90
288
100
%
Carbendazim
4.63
1456
%
Dimethoat
Imidacloprid
3: MRM of 1 Channel ES+
TIC
1.45e3
Area
2: MRM of 1 Channel ES+
TIC
5.64e3
Area
Metsulfuron -methyl
0
Eq_STD1pg_A7_002
3.78
692
100
%
1
2.50
1: MRM of 1 Channel ES+
TIC
3.44e3
Area
Methomyl
5.00
5: MRM of 1 Channel ES+
TIC
2.77e4
Area
4: MRM of 1 Channel ES+
TIC
2.70e3
Area
1
Eq_STD1pg_A7_002
100
6: MRM of 1 Channel ES+
TIC
1.90e4
Linuron
Area
7.50
10.00
12.50
15.00
17.50
Time
20.00
Confirmation with MS—When paired with an Alliance HPLC System, a tandem quadrupole
mass spectrometer provides improved resolution. In this example, the Alliance System used
with the Quattro Premier XE Mass Spectrometer exceeds regulatory requirements for the
identification and quantitation of pesticide residues in wheat samples.
ZQ™—Our versatile single
quadrupole mass spectrometer
for general mass confirmation.
Leading chemistry products
Being an integral part of a world-renowned
organization gives Waters Chemistry products a
powerful advantage few others enjoy. Waters has
a wide range of columns, Solid-Phase Extraction (SPE)
cartridges and 96 well plates, Life Science solutions,
and many supplies and accessories.
configurations, guaranteeing unsurpassed
Increase capacity—With the Alliance System’s optional solvent
tray, you can use higher capacity solvent bottles.
Quattro Premier™ XE—Achieve
high performance and low
detection limits with our compact,
easy-to-use tandem quadrupole
mass spectrometer.
LCT Premier™—An oa-Tof MS unit
with the resolution/sensitivity to identify
components in complex mixtures.
HPLC Columns
Waters’ HPLC column portfolio extends from the
XBridge™ columns—our second-generation hybrid
particle technology—to SunFire for state-of-the-art
silica phases, to Atlantis®, the industry’s solution to
polar compound retention; as well as established products
such as Symmetry® and Intelligent Speed (IS) columns.
versatility and flexibility
Compounds
1. Doxylamine
2. Benzamide
3. Hydroxyisophthalic Acid
4. Doxepine
5. Flavone
6. Fenoprofen
3
2
4
5
6
1
pH 2
4
3
5
6
2
1
pH 7
3
2
5
6
4
2996 Photodiode Array (PDA)
pH 12
1
Detector—With chromatographic
and spectral sensitivity for
0.00
1.00
2.00
3.00
4.00
analytical and preparative
separations.
5.00
6.00
7.00
8.00
Minutes
Exploring the extremes—Photodiode Arraybased Alliance HPLC Systems deliver nocompromise performance that allows you to get
the most out of your column chemistries. Waters
2487 Dual Wavelength
Absorbance Detector—
The most sensitive UV/Vis
absorbance detector on
the market.
XBridge™ columns are unique in their ability
to withstand aggressive conditions (even at pH
12) allowing complete method development
flexibility, while maintaining the highest
efficiency values associated with silica columns.
1.00
AU
Symmetry ® C18 3.9 x 150 mm, 5 µm
Isocratic
0.00
Check product efficacy—Reliably complete
AU
your dissolution testing for multiple active
1.00
ingredients in less time with an Alliance
Intelligent Speed (IS) columns.
1.00
AU
Dissolution System combined with
Symmetry ® C18 4.6 x 20 mm IS™, 3.5 µm
Isocratic
0.00
Symmetry ® C18 4.6 x 20 mm IS™, 3.5 µm
Gradient
0.00
0.00
1.00
2.00
3.00
Minutes
4.00
5.00
6.00
2414 Refractive Index Detector—
for quantitating low concentrations
With sensitivity, stability for HPLC
of target compounds.
4.00
6.00
8.00
10.00
12.00
14.00
16.00
22.00
Indeno(1,2,3-cd)pyrene - 23.841
Dibenzo(a,h)anthracene - 21.279
20.00
Benzo(g,h,i)perylene - 22.130
Benzo(k)fluoranthene - 18.286
18.00
Benzo(a)pyrene - 19.248
Benzo(b)fluoranthene - 17.203
Benze(a)anthracene - 15.510
Chrysene - 15.987
Fluoranthene - 13.417
Pyrene - 14.057
Phenanthrene - 10.729
Acenaphthene - 8.588
Naphthalene - 5.980
Fluorescence Detector—Sensitivity
Fluorene - 9.206
Fluorescence (EU)
2475 Multi Wavelength
Anthracene - 12.338
Six Injections
Retention Times <0.1% RSD
Peak Areas <1.2% RSD
24.00
26.00
Minutes
and GPC analyses.
Improving the environment—Confidence in your Polycylic
Aromatic Hydrocarbons (PAH) results will increase after
you combine the unwavering gradient reproducibility
of the Alliance HPLC with our high-sensitivity,
programmable, 3D fluorescence detection systems.
432 Conductivity Detector—
With a five-electrode flow cell
design, reduces noise, enhances
sensitivity, and allows direct and
indirect conductivity detection.
1600.00 Soy Oil vs. Canola Oil
Green = Canola Oil
1400.00 Black = Soy Oil
1200.00
LS Units
1000.00
800.00
600.00
2420 Evaporative Light
Scattering Detector (ELSD)—
Can detect molecules that
lack UV/Vis chromophores.
2465 Electrochemical Detector—
400.00
With DC, pulse, and scan
200.00
0.00
0.00
10.00
20.00
30.00
40.00
50.00
60.00
70.00
80.00
Minutes
operation modes for analyzing
a wide range of compounds.
Ensuring quality—Be confident about the quality
of your raw materials and finished product. In a
single run, you can test for adulterated materials
earlier in your process, such as in vegetable oil
origin and purity, shown above.
Sample Preparation
Waters has a comprehensive solution for a wide range of
sample preparation challenges, regardless of your preferred
sample preparation technique. Our sample preparation
Sample vials
products include Oasis®, the modern polymeric SPE products;
Waters’ line of HPLC vials and septa are certified for cleanliness
Sep-Pak®,
by both LC and GC testing to ensure that you get your analysis
the traditional silica SPE products; and
the innovative protein precipitation plates.
Sirocco™,
right the first time.
Solvent management Waters Alliance HPLC Systems employ a simplified fluidic pathway that utilizes a serial flow path with only two check valves—
reducing complexity while enhancing overall ruggedness—across a flow range that covers two orders of magnitude.
• Dependable performance—Due to a simplified serial flow path with two check valves
• Automatic solvent compressibility—Provides worry-free solvent delivery for highly reproducible retention times—with any solvents
• Fixed gradient volume—Ensures consistent, predictable chromatography—regardless of system back pressure
• Flexible flow rate—Programmable without hardware modifications, across a flow range of 50 microliters to 5 milliliters per minute
• Improved uptime—Standard equipment includes an automated, active plunger seal wash
The Alliance HPLC System’s solvent manager
Independently driven pistons achieve smooth
solvent flow—without pulse dampeners—
reducing gradient volume
Dual pressure transducers enable
true pulse-free solvent flow
Ultra-high efficiency vacuum
degassing technology
Precise solvent blending
delivers highly reproducible
gradients
Tool-free routine maintenance on
plungers/seals/seal-wash seals
Simplified flow path with only
two inlet check valves
Sample management The Alliance HPLC System’s sample management design ensures precise sampling—delivering
predictable peak areas, and thus more consistent results—thanks to:
• Five independent carousels—Run samples in one carousel while prepping the sequence in the next;
new samples/carousels can be added without disturbing the ongoing queue
• Sampling routines—For automated reference, peak addition or automated pre-column
derivitization procedures
• Reproducible injections—Select from one to hundreds of microliters, compatible with any
analytical-scale HPLC column chemistry
• Variable volume injections—No need to change loops for analytical methods
• Programmable needle injection—Adjustable height accommodates various
vial geometries (such as different vial bottom thicknesses)
• Programmable syringe draw-rate—Optimizes sampling performance
with various solvent viscosities
• Carryover management—Controlled via a programmable needle wash
cycle and an active needle wash solution
• Consistent sample environment—Samples are kept at a constant temperature
and shielded from exposure to ambient light
• Simplified routine maintenance—The seal wash frit and injection
syringe are easy to access and change without tools
Flexible sample management—Built-in functions, such
as “Auto Add,” allow users to make sample injections
that are composed of aliquots from multiple vials, one
of which is the sample vial. A delay time may be preset
to allow the sample to passively mix prior to injection.
Support at your fingertips
The Alliance Interactive System Support CD ships with every Alliance HPLC System and
contains basic troubleshooting and maintenance information to help keep your Alliance
The CD contains 70 minutes of video clips showing you
Austria 43 1 877 18 07
step-by-step and easy-to-follow maintenance procedures,
Australia 61 2 9933 1777
detailed troubleshooting information, tools to help you
Belgium and Luxembourg
document your methods or system set-up, guidelines for
optimum performance, and links to the Waters website for
fast access to additional information, support, and training
courses for the Alliance Systems.
715000116 Rev. 1 SD
Sales Offices:
© 2003 Waters Corporation
System running at its best.
Alliance Interactive System Support CD
for 2695 Separations Module
Minimum system requirements: Windows 98, 2000, or XP,
Windows Media Player 7 or higher, Pentium III (500Mhz)
processor or higher, 64 MB RAM or higher,
speakers and sound card
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your laboratories perform at their best.
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• Instrument Services extend and enhance the standard limited warranty you receive when
you buy a Waters instrument, delivering the value you need to avoid costly and timeconsuming system downtime.
• Software Services include enterprise implementation and validation services, training, and
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WATERS CORPORATION
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T: 1 508 478 2000
F: 1 508 872 1990
www.waters.com
Waters, Alliance, Atlantis, NuGenesis,
Oasis, Sep-Pak, and Symmetry are registered
trademarks of Waters Corporation. ChromaLynx,
eLab Notebook, Empower, FractionLynx, Intelligent
Speed, IS, LCT Premier, MarkerLynx, MassLynx, MetaboLynx,
OpenLynx, PerformancePLUS, ProfileLynx, ProteinLynx, QuanLynx,
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of Waters Corporation. All other trademarks are the property of their respective owners.
©2005 Waters Corporation Printed in the U.S.A. September 2005 720000370EN LW-MD