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Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns
Published on: April 26, 2016
An improved method for determining free amino acids by RP-HPLC/DAD with o-phthalaldehyde derivatization: Method
Viviane Maria de Sousa Fontes1, Tatiana Colombo Pimentel2, Ana Beatriz Martins da Silva3
1Postgraduate Program in Food Science and Technology, Federal University of Paraíba, João Pessoa, Paraíba, Brazil.
Abstract:
Rapid methods for amino acid determination are desired to reduce running times, and the main factors involved in the rapid separation of these compounds in HPLC are the columns, solvents, and gradient. The present study refers to a method optimization to rapidly analyze 19 amino acids in RP-HPLC/DAD with pre-column derivatization using o-phthalaldehyde. To evaluate the method's robustness, Indian Pale Ale-IPA beers and wines from the San Francisco Valley-SFV and Chapada Diamantina-CHD, Brazil, were analyzed. The method showed acceptable linearity (R2 > 0.992), precision (CV < 3.96%), recovery (74.2-113%), detection limits (<0.56 mg/L), and quantification limits (<3.62 mg/L) and separation gradient in 18.5 min. IPA beers were discriminated from wines by arginine, asparagine, tryptophan, alanine, phenylalanine, and tyrosine contents. Furthermore, CHD wines were discriminated from SFV wines by asparagine, glutamine, arginine, cystine, tyrosine, and leucine. In conclusion, a rapid method for amino acid determination was optimized and validated for wines and beers.
