Citation: YANG Qian, XIAO Bing-Kun, YANG Jian-Yun, HUANG Rong-Qing. Determination of 18 Kinds of Amino Acids Based on A Mixed-mode Retention Mechanism by High Performance Liquid Chromatography-Mass Spectrometry[J]. Chinese Journal of Analytical Chemistry, ;2019, 47(4): 627-633. doi: 10.19756/j.issn.0253-3820.181599 shu

Determination of 18 Kinds of Amino Acids Based on A Mixed-mode Retention Mechanism by High Performance Liquid Chromatography-Mass Spectrometry

Institude of Radiation Medicine, Academy of Military Medical Sciences, Beijing 100850, China

Received Date: 18 September 2018
Revised Date: 28 December 2018

Amino acids are usually analyzed by derivatization or adding ion-pair reagents. In this work, a new method was established base on a mixed-mode retention mechanism (reversed-phase and cation-exchange) to determinate 18 kinds of amino acids, using simple mobile phase without the ion-pair and derivatization. In this method, 18 kinds of amino acids could be retained on a OSAKA SODA CAPCELL PAK CR 1:4 (150 mm×2.1 mm, 5 μm; SCX:C₁₈=1:4) column with a mobile phase of 0.1% formic acid(A)-acetonitrile (0.1% formic acid) (B) - 50 mmol/L ammonium formate (C) using a gradient elution program. Mass spectrometry (MS) detection was performed with multiple reaction monitoring (MRM) using positive electrospray ionization. The relative standard deviations (RSDs) of peak area ranged from 0.7% to 5.9%. The correlation coefficients (R²) of 18 kinds of compounds were greater than 0.993. And the recoveries of serum and urine samples from SD rats ranged from 92.2% to 113.6%. This method was proved to be simple with good reproducibility. It provides a new method and idea for determination of amino acids in biological samples or other complex matrices.
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