ACQUITY UPLC M-Class System with 2D Technology
Expand the dynamic range of your analysis with 2D separations
Until recently, 2D-LC was considered time-consuming and difficult to accomplish. The ACQUITY UPLC M-Class System with 2D Technology has streamlined the 2D-LC separation process with highly intuitive menu-driven method setup, standardized separation chemistries, and intelligent valve operation.
The ACQUITY UPLC M-Class System with 2D Technology expands the use of sub-2-micron particles to achieve high peak capacity separations for peptides. This innovative system effectively uses two-dimensional (2D) UPLC for better chromatographic resolution of complex proteomic samples by using a dual reversed-phase (RP) approach.
ACQUITY UPLC M-Class System with 2D Technology
Specifications
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Maximum operating pressure |
15,000 psi |
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pH range |
2 to 10 |
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Operating flow rate range |
200 nL/min to 100 μL/min without flow splitting |
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Sample flow path |
Non-ferrous |
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Injection volume |
0.1 µL to 100.0 µL |
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Sample compartment |
4–40 ˚C |
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Pump options |
Binary |
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Column tracking |
None |
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Column capacity |
Single column, 75 µm to 4.6 mm internal diameter (ID); up to 250 mm in length |
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Maximum sample capacity |
Up to 21 plates (with Sample Organizer) |
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Detector options |
PDA Detector TUV Detector Mass Spectrometers |
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External control |
MassLynx Software |
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Physical specifications (µBSM, µSM-FL, TVM) |
Width: 34.3 cm (13.5 in.) Height: 69.7 cm (27.4 in.) Depth: 71.2 cm (28.0 in.) |
Overview
- High-resolution in both dimensions by exploiting the wide-ranging ionic and hydrophobic structure of peptides
- Better protein identifications, quantification, and sequence coverage
- Improved separation, method generation wizards, and enhanced data algorithms
Recommended Use: For better chromatographic resolution of complex proteomic samples.
Features Header
Expanding the impact of comprehensive 2D
Conventional comprehensive 2D-LC uses ion exchange (IEX) followed by reversed-phase (RP). Any IEX approach will use salt-containing buffers that can cause ionization background and fouling problems if they enter a mass spectrometer (MS). Since IEX separations are based solely on the charge of the peptide, the IEX dimension often results in poor chromatographic resolution with peptides appearing in multiple fractions, making data interpretation difficult.
The improved 2D approach of ACQUITY UPLC M-Class System with 2D Technology uses RP at pH 10 in the first dimension, followed by RP at pH 2 in the second dimension – for results that far exceed those of conventional IEX methodologies.
Secure identifications for best-in-class reproducibility
As a function of the ACQUITY UPLC M-Class System with 2D Technology’s 2D-LC resolution, your lab can see an increased number of secure identifications, including both reproducible and incremental identifications of proteins based on 2D-LC. This results in better protein identifications, quantification, and sequence coverage.
High resolution in both dimensions
Using IEX as the first separation dimension often results in poor resolution and bleed between fractions. Using a highly resolving support in the first step provides your lab with better information in the last step. With the reversed-phase support at pH 10 followed by a sub-2-micron separation at lower pH with ACQUITY UPLC M-Class System with 2D Technology, you can obtain useful, orthogonal separations that yield high quality data.
Increased resolution equals better protein identifications
In this separation, the power of comprehensive 2D-LC for resolution of complex proteomic samples is demonstrated by this tryptic digest of E. coli. Using the ACQUITY UPLC M-Class System in one dimension, 0.5 µg were loaded on-column to resolve and identify 365 proteins from 2874 peptides. Identification of 7661 and 9415 peptides from the 5-step and 10-step separations respectively were obtained by adding 2.5 µg to the 2D-LC system, resulting in greater protein coverage with high confidence in results.
With 2D-LC and the ACQUITY UPLC M-Class System, labs can identify twice as many proteins more securely, recover more peptides, and achieve greater protein coverage with high confidence in results.
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