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Abstract
This chapter focuses on the chromatography of amino acids (AAs) and peptide analysis. The approach to AA analysis depends on the origin of a specimen. Specimens may be divided into three classes: (1) mixtures of AAs derived from peptide hydrolysis, (2) AAs derived from peptide sequencing, and (3) AAs present in some biological fluid. The major task in AA analysis is the determination of free AAs in biological fluids. A specimen may contain many unknown substances that can affect the reliability of the analysis. Conventional AA chromatography is currently performed mainly on octadecyl silica by reversed-phase, high-performance liquid chromatography (RP-HPLC) with water/organic solvent gradients. The most common method for peptide analysis utilizes a HPLC procedure with a column of 25-cm length and 4.1-mm ID, packed with a 5-μm, wide-pore octadecylor octyl-bonded stationary phase, eluted with a water/acetonitrile gradient and 0.1–0.2% of trifluoroacetic acid (TFA) as ion-pairing and solubilizing agents.
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