ACQUITY UPLC M-Class System with 2D Technology

Expand the dynamic range of your analysis with 2D separations.

The ACQUITY UPLC^® M-Class System with 2D Technology expands the use of sub-2-micron particles to achieve high peak capacity separations. 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.

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

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ACQUITY UPLC M-Class System w/2D Technology

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Comprehensive 2D - Expanding the impact.

Conventional 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.

This improved 2D approach uses RP at pH 10 in the first dimension, followed by RP at pH 2 in the second dimension for results that far exceeds those of conventional IEX methodologies.

Secure Identifications for Best-in-Class Reproducibility

An increased number of secure identifications is shown as a function of 2D-LC resolution. Both reproducible and incremental identifications of proteins based on 2D-LC can be seen.

High resolution in both dimensions.

When ion exchange is used as the first separation dimension, poor resolution and bleed between fractions could occur. Using a highly resolving support in the first step provides you with better information in the last step. A reversed phase support at pH 10, followed by a sub-2-micron separation at lower pH, useful, orthogonal separations that yield high quality data is obtained.

Increased resolution equals better protein identifications.

In this separation, the power of 2D-LC for resolution of complex proteomic samples is demonstrated by this tryptic digest of E. coli. Using ACQUITY^® UPLC M-Class 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.

The 2D-LC enabled

Identification of twice as many proteins more securely based
Recovery of more peptides
Greater protein coverage with high confidence in results

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