Waters ACQUITY UPLC M-Class System with 2D Technology

ACQUITY™ UPLC M-Class System with 2D Technology

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The ACQUITY UPLC® M-Class System provides reliable high resolution chromatographic separations optimized for columns from 75 μm to 1.0 mm internal diameter. The system’s high pressure, direct (non-split) solvent and sample management system is optimized to minimize dispersive and adsorptive losses. The new sample manager and complementary valve and trapping management module delivers maximum sample recovery with superior chromatographic resolution through efficient use of sub-2-µm particles for a variety of system and column configurations to provide extensive application flexibility.

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.

Key Features

• 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.

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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