Quantification of underivatized amino acids in solid beverages using high-performance liquid chromatography and a potentiometric detector

Abstract

The simultaneous quantification of amino acids (AAs) in solid beverages without prior derivatization was explored by high-performance liquid chromatography (HPLC) coupled to a potentiometric detector. Included were threonine, leucine, methionine, phenylalanine, and histidine. The potentiometric detector was made consisting of a copper(II)-selective electrode based on a polyvinyl chloride (PVC) membrane, and the potential changes in the detector were determined according to the coordination interactions between cupric copper ions released from the inner filling solution of the electrode and AAs. Conditions were optimized for effective separation and sensitive detection. Fundamental characteristics such as linearity, limits of detection, limits of quantitation, accuracy, precision, and robustness were validated experimentally. The calibration curves showed a linear relationship between peak heights and the injection concentrations of the AAs. The detection limits down to the sub-micromolar range were achieved under isocratic conditions, outperforming ultraviolet detection. The copper(II)-selective electrode had a minimum lifetime of one month. Some real samples were examined to further demonstrate the feasibility of the proposed approach. The measurement results obtained by the present method were in good agreement with those obtained by the HPLC-mass spectrometry (MS), indicating that the combined HPLC-potentiometric method is a potential option for quantifying AAs.