ACQUITY PREMIER Solution for Biomedical Research

 

Expanding the coverage of biomarkers classes and detecting biologically significant low-level markers are essential in biomedical research. Cellular markers of disease and key signaling molecules like organic acids, phosphorylated and carboxylated lipids and metabolites, or phosphorylation of proteins are often challenging to detect and accurately quantify because they readily adsorb to metal surfaces and are lost in the analytical process due to analyte/surface interactions. Innovative ACQUITY PREMIER solution featuring MaxPeak™ High Performance Surfaces significantly reduces these analyte/surface interactions and increases the possibility of novel discoveries and a deeper understanding of the biology.

Non-specific binding or adsorption leads to poor chromatographic peak shape, low recovery, reduced sensitivity, and even complete loss of analytes. Long system equilibration and use of chemical additives to passivate the surface are common practice to reduce analyte/surface interactions and improve chromatographic peak shape and recovery. Waters ACQUITY PREMIER solution featuring MaxPeak™ High Performance Surfaces is designed to minimize such non-specific adsorptions expands the coverage of biomarker classes, improve chromatographic peak shape, separation, reproducibility, and sensitivity of biologically important metabolites, lipids, and phospho-peptides.

Application Notes
  • ACQUITY PREMIER LC Technology Significantly Improves Sensitivity, Peak Shape, and Recovery for Phosphorylated and Carboxylate Lipids
  • Improved Analytical Sensitivity and Chromatographic Peak Shape for the Quantification of TCA Cycle Analytes in Human Plasma using the ACQUITY PREMIER System Solution
  • Maximizing Phosphopeptide Recovery in LC-MS Studies with MaxPeak High Performance Surfaces Technology

Phosphorylated and Carboxylate Lipid Analysis
Phosphorylated and carboxylate lipid species are metal sensitive and can readily absorb to stainless steel surfaces within the flow path of UPLC™ systems leading to poor peak shape, low recovery and reduction in sensitivity. The ACQUITY PREMIER solution provides a highly reliable method for the analysis of LPA, PA, LPS and PS lipid molecular species that significantly improves sensitivity, peak tailing and recovery of phosphorylated and carboxylate lipids. The developed method enables increased lipidomics coverage and detection of detection of low level signaling lipids such as phosphatidylinositol phosphates with mono, di and tri phosphates, ceramide-1-phosphate and Sphingoid base 1-phosphates.

ACQUITY PREMIER solution improves the peaks shape, peak area, and sensitivity for Lysophosphatidic acid LPA (16:0/0:0) 
ACQUITY PREMIER solution improves the peaks shape, peak area, and sensitivity for Lysophosphatidic acid LPA (16:0/0:0)
 ACQUITY PREMIER solution eliminates peak tailing, improves peaks shape, peak area, and sensitivity for Lysophosphatidylserine LPS(18:1/0:0)
ACQUITY PREMIER solution eliminates peak tailing, improves peaks shape, peak area, and sensitivity for Lysophosphatidylserine LPS(18:1/0:0)

 

 

Improved Sensitivity and Chromatographic Peak Shape for the Quantification of TCA Cycle Analytes
The analysis of TCA cycle and related metabolites in human plasma is challenging due to their small, polar nature with many isobars, wide range of concentrations in biological samples, and finally, metal sensitivity. The ACQUITY PREMIER solution mitigates analyte interactions with metal to improve peak shape as well as sensitivity without full system passivation with strong acids or chelating additives in the mobile phase. The simple mixed-mode anion exchange separation using the ACQUITY PREMIER CSH Phenyl-Hexyl column allows for fast separations and analysis of the TCA cycle and related metabolites in human plasma with great sensitivity.

 Separation of TCA cycle metabolites from plasma using the ACQUITY PREMIER solution shows an increase in peak area for iso-citric and citric acids, reduced peak tailing for malic acid, and detection of low-level metabolite, 3-phosphoglyceric acid, often missed by conventional LC.
Separation of TCA cycle metabolites from plasma using the ACQUITY PREMIER solution shows an increase in peak area for iso-citric and citric acids, reduced peak tailing for malic acid, and detection of low-level metabolite, 3-phosphoglyceric acid, often missed by conventional LC.

 

 

Phosphopeptide Recovery Enhancements with MaxPeak High Performance Surfaces Technology
MaxPeak High Performance Surface (HPS) present in ACQUITY PREMIER solution limits the metal ion interaction with the phosphate group present in the peptide and dramatically improves chromatographic peak shape, recovery, and mass spectrometry response phosphopeptides.

 Analysis of tryptic digest of the phosphorylated protein, Alpha Casein, using the ACQUITY PREMIER solution, enables the detection of new phosphopeptides that are not seen the conventional LC, and expands the coverage in biomarker discovery.
Analysis of tryptic digest of the phosphorylated protein, Alpha Casein, using the ACQUITY PREMIER solution, enables the detection of new phosphopeptides that are not seen by the conventional LC, and expands the coverage in biomarker discovery.