Assay for Characterizing Adsorption-Properties of Surfaces (APS)

Area of Science:

• Analytical Chemistry
• Biochemistry

Background:

• Analyte adsorption to surfaces during sample preparation is a significant challenge in quantitative analysis, particularly in omics studies.
• Existing methods like internal standards are insufficient for minimizing overall analyte loss across diverse molecules.
• Understanding and quantifying adsorption interactions is crucial for improving analytical accuracy.

Purpose of the Study:

• To develop and validate a novel assay for qualifying and quantifying adsorption of molecules to surfaces (APS).
• To assess the adsorption properties of various vial surfaces using a comprehensive peptide mixture.
• To provide a method for approximating analyte loss during sample preparation.

Main Methods:

• Utilized liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based differential quantitative analysis.
• Employed a reference mixture of thousands of tryptic peptides with diverse chemical characteristics.
• Investigated adsorption properties of different vial types, including conventional polypropylene, low-protein-binding polypropylene, and glass vials.

Main Results:

• Demonstrated significant adsorption of hydrophobic peptides to conventional polypropylene vials.
• Observed minimal peptide adsorption to low-protein-binding polypropylene vials.
• Identified substantial peptide adsorption to glass vials, primarily driven by electrostatic interactions.

Conclusions:

• The developed assay effectively characterizes the adsorption properties of different surfaces.
• The assay can approximate analyte loss during sample preparation, aiding in method optimization.
• Findings highlight the impact of surface chemistry on analyte recovery and suggest appropriate vial selection for omics studies.