Korean J Physiol Pharmacol.  2012 Oct;16(5):355-360. 10.4196/kjpp.2012.16.5.355.

Simultaneous Determination of Glutamate, Glycine, and Alanine in Human Plasma Using Precolumn Derivatization with 6-Aminoquinolyl-N-hydroxysuccinimidyl Carbamate and High-Performance Liquid Chromatography

Affiliations
  • 1Institute of Pharmacy and Department of Pharmacology, School of Pharmacy, Binzhou Medical University, Yantai 264003, China. lqingzhong@yahoo.com.cn
  • 2Department of General Surgery, Yantai Yuhuangding Hospital, Yantai 264003, China.
  • 3Department of Quality Inspection, Hubei Provincial Bafeng Pharmaceuticals & Chemicals Share Co., Ltd, Hefeng 445800, China.

Abstract

A simple, sensitive and reproducible high-performance liquid chromatography (HPLC) method has been validated for determining concentrations of glutamate, glycine, and alanine in human plasma. Proteins in plasma were precipitated with perchloric acid, followed by derivatization with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC). Simultaneous analysis of glutamate, glycine, and alanine is achieved using reversed-phase HPLC conditions and ultraviolet detection. Excellent linearity was observed for these three amino acids over their concentration ranges with correlation coefficients (r)>0.999. The intra- and inter-day precision were below 10%. This method utilizes quality control samples and demonstrates excellent plasma recovery and accuracy. The developed method has been successfully applied to measure plasma glutamate, glycine, and alanine in twenty volunteers.

Keyword

Amino acids; AQC; HPLC; Plasma; Ultraviolet detection
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