Liquid Chromatography-Mass Spectrometry (LC-MS) Analysis of Bioactive Peptides

Abstract

Liquid chromatography (LC) is the most widely used separation technique for peptide analysis. This chapter discusses different steps in the mass spectrometry (MS)-based peptidomic analysis are discussed, including sample preparation, LC separation, detection, and data-acquisition using MS as well as data analysis. In order to investigate the peptide composition in complex food or biological matrices, an adequate sample preparation is required as matrix components can interfere with the peptide LC-MS signal. Bioactive peptides, originating from food proteins after digestion or hydrolysis, can be identified using different techniques, after which confirmatory in vitro and in vivo functionality studies may be carried out with the identified synthetic peptides. The potential of nano-LC-MS in peptide food analysis was demonstrated by the identification of the bioactive peptides produced during in vitro gastrointestinal digestion of soybean seeds and soy milk. When using liquid chromatography for peptide separation, different stationary phases can be applied. Most peptide detection and quantification occurs via ultraviolet detection and/or mass spectrometry.