Agilent InfinityLab 2D-LC Solution
SOLVE ANY SEPARATION CHALLENGE
WITH MAXIMUM EFFICIENCY
INTRODUCTION
SOLVE ANY SEPARATION CHALLENGE
WITH MAXIMUM EFFICIENCY
Solve your most complex separation challenges with the most powerful and easiest-to-use 2D-LC
solution. The Agilent InfinityLab 2D-LC solution puts you on the fast track to unmatched separation
power. What‘s more, you can match separation performance to your needs and sample complexity.
At the flip of a switch you can select between 1D UHPLC and different levels of 2D-LC power.
2D-LC
2D-LC
Conventional 1D-LC
Conventional 1D-LC
2D-LC Modes
Heart-Cutting
HiRes sampling
Peak capacity
2D-LC
1D-LC
Comprehensive
Analysis time
ANALYTICAL
INSTRUMENT
LABORATORY
Solve your most complex
separation challenges
with unmatched levels
of separation power and
achieve highest efficiency
for any separation task.
Benefit from the full choice
of peak capacity and
resolution by switching
between 1D and all 2D
modes with intuitive
software – but without
manual interaction.
Use a single, fully scalable
system for all your 1D
and 2D separation tasks.
Get up-and-running fast
through dedicated training.
EFFICIENCY
2
EFFICIENCY
EFFICIENCY
COMPLETE SOLUTIONS
The Agilent InfinityLab 2D-LC solution is a truly complete end-to-end solution, combining
superior instruments and columns, smart supplies, intuitive software and dedicated services.
InfinityLab Instruments, Columns and Supplies
Designed to work together in perfect harmony, the Agilent 1290 Infinity II LC combines with Agilent
InfinityLab Poroshell 120 columns to give you unmatched separation power. Agilent InfinityLab Quick
Connect and Quick Turn fittings make column installation an easy task, and Agilent InfinityLab Stay
Safe solvent caps protect you from harmful vapors.
Services and Support
Software and Informatics
Learn how to gain the maximum benefit from
your InfinityLab 2D-LC solution through CrossLab’s
dedicated and intensive training course on
two-dimensional UHPLC.
Separate your most complex samples intuitively
and with ease through OpenLAB´s complete
set of fully flexible one- and two-dimensional
UHPLC capabilities.
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TECHNOLOGY
MATCH SEPARATION POWER
TO APPLICATION NEEDS
The InfinityLab 2D-LC solution gives you easy access to any 2D separation mode. Choose from multiple
heart-cutting and heart cutting, high-resolution sampling, or comprehensive 2D-LC to match separation
power to your application needs. And for less-challenging tasks, simply switch back to 1D UHPLC
without reconfiguring your system.
Heart-cutting and multiple heart-cutting
2D-LC for confident peak-purity determination
Heart-cutting and multiple heart-cutting 2D-LC (MHC 2D-LC) allow
you to combine orthogonal separation and detection systems not
usually considered compatible, providing additional information
about your sample for an extra level of confidence.
First dimension
• Additional separation for dedicated areas in a chromatogram
• Unmatched peak storage in ready-to-use valves and loops
• Ideal for impurity analysis or desalting before MS detection
Second dimension
One-dimensional separations often conceal impurities and prevent you from determining whether a peak is pure. For utmost confidence in your peak-purity determinations,
heart-cutting and multiple heart-cutting 2D-LC allow you to focus on one or more areas and extract additional information about your sample.
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High-resolution sampling 2D-LC
and accurate quantification
Available only from Agilent, high-resolution sampling 2D-LC
(HiRes 2D-LC) does not sacrifice chromatographic resolution in
either dimension for highest separation power and quantification
accuracy in trace-level impurity analysis.
First dimension
• Additional separation for dedicated areas in a chromatogram
• Unmatched resolution in both dimensions
• Ideal for many biopharmaceutical applications
Second dimension
The first-dimension separation gives you valuable performance you do not want to sacrifice. High-resolution sampling 2D-LC solves this challenge by giving you additional separation
power to quantify the trace-level peaks with highest accuracy.
Comprehensive 2D-LC
for highest peak capacity
Comprehensive 2D-LC (LCxLC) achieves maximum
separation power with highest overall peak capacity to give
you a complete overview of the entire sample and determine
each and every component.
• Additional separation for the entire chromatogram
• Highest accuracy and precision
• Ideal for sample screening or identity control
First and second dimension
Standard UHPLC separation of a complex sample, showing
overlapping peaks. Identification of all peaks is not possible.
Comprehensive 2D-LC separation of the same
sample. Highest separation power delivers the
highest level of information about the sample.
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TECHNOLOGY
ACCESS FULL 2D-LC POWER WITH EASE
The InfinityLab 2D-LC solution combines best-in-class UHPLC separation hardware with intuitive
software to give you easy access to the full power of two-dimensional liquid chromatography.
Benefit from highest separation performance with any mode of 2D-LC.
Three simple steps to highest instrument efficiency
The Agilent 2D-LC acquisition
software is based on familiar
OpenLAB methods for UHPLC,
making it a fast and simple task
to set up any 2D-LC method:
1
2
1 Choose the 2D-LC mode
2 Program your 2D gradient
3 Select cuts and/or adjust
gradients
3
Whether you require comprehensive, heart-cutting, multiple heart-cutting, or Agilent´s unique
high-resolution sampling 2D-LC, method setup is straightforward and gives you the flexibility to
design the most challenging two-dimensional separation scheme. Use the Preview window to
adapt the setup to meet your specific demands.
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Simplicity for highest analytical efficiency
two-dimensional liquid chromatography, a 2D-LC valve unique to
Agilent couples the first and second dimensions for easy setup and
straightforward operation. Combining the 2D-LC valve with multiple
heart-cutting 2D-LC valves, also unique to Agilent, provides for
unmatched capabilities to achieve highest resolution.
The Agilent 1290 Infinity II Speed Pumps deliver flow of the highest
accuracy and precision, ensuring highest speed and performance in
both dimensions. Lowest delay volumes enable fastest gradients and
extraordinary pump intelligence ensures identical start conditions
for highest 2D-gradient repeatability. Meeting the precise needs of
First dimension
Second dimension
1290 Infinity II
Diode Array
Detector
Waste
2
3
1
4
1290 Infinity II
High Speed Pump
8
5
1290 Infinity II
Multisampler
6
7
1290 Infinity II
Diode Array
Detector
1 D-Column
1290 Infinity II
High Speed Pump
2 D-Column
1290 Infinity II Multicolumn Thermostat
Waste
Typical setup for comprehensive and single heart-cutting 2D-LC, showing the symmetrical flow paths for the first and second dimensions. Expand your capacity
through straight­forward exchange of the loops for multiple heart-cutting 2D-LC valves.
Agilent 2D-LC valve
• Manufactured with absolutely symmetrical flow paths
• Eliminates any possible switching interference
• Designed for all 2D-LC modes and applications
Agilent multiple heart-cutting 2D-LC valve
• Designed for multiple heart-cutting and high-resolution sampling 2D-LC
• Preconfigured, ready-to-go valves with 12 loops
• Fully integrated in 2D-LC software with live status monitor
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APPLICATIONS
2D-LC VERSATILTY FOR APPLICATION FLEXIBILITY
The InfinityLab 2D-LC solution offers the versatility you need to address the widest variety of
applications. In a single run, you can use the maximum separation power for a complete overview
of the entire sample, or cut out dedicated sections for further separation.
Impurity analysis
The identification and quantification of impurities is one of the most important tasks in the analysis of pharmaceuticals. Targeted analysis
using multiple heart-cutting 2D-LC enables determination of multiple hidden impurities at critical places in the first-dimension separation –
all in a single run.
Download Application Note from www.agilent.com, search for 5991-5643EN.
Second dimension
First dimension
Absorbance [mAU]
100
80
2
60
1
40
1
20
0
6
8
10
12
14
16
18
20
Time [min]
Two out of 11 heart-cuts of the first-dimension separation revealed coeluting impurities when separated in the second dimension.
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2
Determination of polyaromatic
hydrocarbons in petroleum products
Samples of petroleum products are possibly the most complex encountered in any laboratory. The challenges these samples present can be
solved using a highly orthogonal 2D-LC separation system with normal and reversed-phase columns. Multiple heart-cutting 2D-LC enables
separation of the peaks of interest from the high matrix load.
Download Application Note from www.agilent.com, search for 5991-6549EN.
First dimension
Second dimension
mAU
LU
Std 1 ppm
400
PAH 8
7
Petroleum
vacuum residue
Std 1 ppm
Petroleum
vacuum residue
6
5
300
4
3
200
100
5-6
9-10
2-4 8
11-13
15-16 14
7
1
0
0
10
20
PAH 7
2
1D column
backflush
1
0
Time [min]
10
12
14
16
Time [min]
2D-LC analysis of a petroleum vacuum residue sample and a PAH standard mixture, showing one cut as example of the second-dimension analysis for two selected PAHs.
Fingerprint profiling of herbal medicines
Traditional herbal medicine is an extremely complex science, involving hundreds of herbal mixtures that contain thousands of different
substances. For detailed identification, comprehensive 2D-LC enables fingerprint profiling of these diverse mixtures, revealing specific
similarities and differences. The example illustrates the additional power of a second dimension.
Download Application Note from www.agilent.com, search for 5991-5028EN.
Angelica sinensis radix (Dang Gui)
Comprehensive 2D-LC analysis of the traditional herbal medicine Dang
Gui (Angelica sinensis radix), demonstrating the ability of this technique
to deliver fingerprint profiles of complex mixtures in a single run.
For Research Use Only. Not for use in diagnostic procedures.
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APPLICATIONS
2D-LC POWER FOR BIOPHARMA ANALYSIS
The high separation power of 2D-LC makes it ideal for the analysis of highly complex
biopharmaceuticals. Whether you need comprehensive screening or detailed determination
of specific peptides, the InfinityLab 2D-LC solution helps you solve the toughest biopharma
application challenges.
Screening of monoclonal antibodies
The analysis of monoclonal antibodies presents specific challenges due to the complexity of both the analytes and the matrixes. The
determination of differences between stressed and non-stressed antibodies is especially challenging because of multiple overlapped peaks
that can only be distinguished with orthogonal separation techniques. Comprehensive 2D-LC provides an ideal tool for initial screening prior
to more detailed analysis.
Download Application Note from www.agilent.com, search for 5991-2880EN.
Comprehensive 2D-LC contour plot for the analysis of a tryptic digest of pH-stressed trastuzumab. For Research Use Only. Not for use in diagnostic procedures.
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Determination of impurities in
monoclonal antibodies
The technique of high-resolution sampling 2D-LC is unique to Agilent and enables highly specific selection from the first-dimension
separation for second dimension analysis. This technique combines the advantages of comprehensive and multiple heart-cutting 2D-LC
to resolve even the most complex areas of a chromatogram.
Download Application Note from www.agilent.com, search for 5991-6906EN.
Second dimension
First dimension
Absorbance [mAU]
1
500
2
3
4
5
6
7
Absorbance [mAU]
120
8
100
400
80
300
60
200
40
100
20
0
0
2.5
3
Time [min]
0
1
2
3
Time [min]
In this example, four peaks can be determined by applying 2D-LC in just a five-second cut from the 1D chromatogram. High-resolution sampling 2D-LC maintains the 1D separation
for utmost resolution in the entire 2D-LC system and ensures reduction of further possible interferences. For Research Use Only. Not for use in diagnostic procedures.
MS analysis of monoclonal antibodies
using 2D-LC for eluent desalting
Successful characterization of monoclonal antibodies requires complex chromatographic systems such as hydrophobic interaction (HIC),
strong cation exchange (SCX), or weak anion exchange (WCX) that deploy high levels of buffers or salts. Multiple heart-cutting 2D-LC
provides an easy way to desalt the eluent, enabling direct connection to mass-selective detectors.
Download Application Note from www.agilent.com, search for 5991-6673EN.
2D chromatogram
1D chromatogram
3
mAU
300
4
5
6
7
8
9
10
11
40
200
20
100
0
14
16
18
Time [min]
0
1
2D mass spectra
Counts ×10 4
G0F/G1F
6
147,243.98
5
G0F/G2F or (G1F)2
4
147,406.86
3 G0F/G0F
G1F/G2F
2 147,082.00
147,568.49 147,997.30
1
147,739.39
148,152.01
0
147,000
147,400
147,800
148,200
Deconvoluted mass [amu]
Even when desalting prior to MS analysis is not the primary objective, multiple heart-cutting 2D-LC gives the flexibility to run longer 2D gradients that are often required
for the analysis of complex biological samples. For Research Use Only. Not for use in diagnostic procedures.
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Learn more
We have compiled a comprehensive collection of 2D-LC applications, including examples of
chemical and food analysis, lipidomics and metabolomics, herbal medicine analysis, and the
combination of achiral and chiral separations. We also offer dedicated 2D-LC courses for
beginners and experienced chromatographers.
www.agilent.com/chem/2D-LC
Buy online
www.agilent.com/chem/store
Find an Agilent customer center
www.agilent.com/chem/contactus
USA and Canada
1-800-227-9770
agilent_inquiries@agilent com
Europe
[email protected]
Asia Pacific
[email protected]
This information is subject to change without notice.
© Agilent Technologies, Inc. 2017
Published in the USA, January 1, 2017
5991-7226EN