Literature DB >> 16741240

Validation of a reversed-phase HPLC method for quantitative amino acid analysis.

Maria Paola Bartolomeo1, Federico Maisano.   

Abstract

A semi-automated method for amino acid derivatization and analysis has been validated for use in analysis of protein biopharmaceuticals. The method includes protein hydrolysis, o-phthalaldehyde derivatization, and reversed-phase high-performance liquid chromatography analysis in a general-purpose UV-visible high-performance liquid chromatography system. Amino-acid derivatization is performed automatically by the high-performance liquid chromatography autosampler right before injection. The required validation parameters, i.e., specificity, linearity, accuracy, precision, limit of detection, and limit of quantification, were studied for bovine serum albumin and for a recombinant human Fab fragment. The method can be employed as an absolute quantification method for determination of extinction coefficients of recombinant proteins.

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Year:  2006        PMID： 16741240      PMCID： PMC2291777     

Source DB:  PubMed          Journal:  J Biomol Tech        ISSN： 1524-0215


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