Device and method for online deduction of amino acid disulfide and determination of amino acid

Abstract

The invention provides a device and a method for online deduction of amino acid disulfide and determination of amino acid. The device mainly comprises a high-pressure pump, a high performance liquid chromatographic column, an online reduction column, and a chemiluminescent detector. The basic technical route is as follows: a amino acid solution to be determined enters the online reduction column after being subjected to high performance liquid chromatographic column separation; and the amino acid disulfide in the solution is restored into corresponding mercaptoamino acid by tri(2-propyloic) phosphine complex crystal of zinc (II) in the online reduction column, and then enters the chemiluminescent detector to be subjected to online determination of the amino acid. The online reduction device is simple and convenient in detection method, rapid in reduction process, high in reduction efficiency, good in repeatability, and wide in application prospect, and newly restored mercaptoamino acid is prevented from being oxidized into the amino acid disulfide again in sample treatment and analysis processes.

Description

technical field [0001] The invention relates to a device and method for rapidly on-line reduction of amino acid disulfide and determination of amino acid. The device and method are suitable for rapid online determination of the content of sulfhydryl amino acid and amino acid disulfide in urine or plasma samples. Background technique [0002] The content of thiol amino acids and amino acid disulfides in biological fluids is closely related to human health. Through the determination of mercapto amino acid and amino acid disulfide content, the health of the human body can be monitored in real time. The method currently used is to reduce the amino acid disulfide to the corresponding thiol amino acid off-line, and then measure the content of various thiol amino acids, thereby determining the content of various amino acids in the sample. The literature Muhammad I.K., Zafar I. Simultaneous determination of ascorbic acid, aminothiols, and methionine in biological matrices using ion...

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Problems solved by technology

This off-line operation is not only a waste of time, but mercapto amino acids are easily oxidized to amino acid disulfides during sample processing and subsequent analysis

Documents James L., James D. The determination of disulphide species within physical fluids. Trends Anal. Chem., 2012, 21:807-815. reported the direct simultaneous online determination of thiol amino acids and amino acid disulfides by electrochemical or mass spectrometry methods, This determination method is relatively simple in procedure, but the electrode stability of the electrochemical method is poor, and it is easily affected by interfering substances, resulting in a decrease in sensitivity; the mass spectrometry method is complex and expensive

At present, most of the determination of sulfhydryl amino acids and amino acid disulfides still use indirect methods. These methods have many steps and are time-consuming.

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