Method for separating and measuring acetylcysteine enantiomers

Abstract

The invention provides a method for separating and measuring acetylcysteine enantiomers. In the method, before acetylcysteine is added in a chromatographic column, a derivatization reagent is used to perform derivatization, wherein the derivatization reagent is N(alpha)-(5-fluoro-2,4-dinitrophenyl)-L-amino acid compound. The method for separation and measurement combines the high performance liquid chromatography (HPLC) or high performance liquid chromatography-mass spectrum (HPLC-MS), and the used chromatographic column uses octadecylsilane chemically bonded silica as filler. The method for separation and measurement comprises the following steps: (1) taking acetylcysteine or a preparation with acetylcysteine, dissolving in low-concentration acid solution, adjusting the pH value of the mixed solution to 6.0-8.0 with alkaline solution to obtain a sample solution for testing; (2) mixing the acetylcysteine solution with 1mol / L of carbonate solution, adding N(alpha)-(5-fluoro-2,4-dinitrophenyl)-L-amino acid compound to mix evenly; (3) reacting the solution obtained by the step (2) at 40-60 DEG C in a dark place, adding hydrochloric acid solution after the reaction; (4) adding the solution obtained by the step (3) in a drying solution with the prestored phosphorus pentoxide and potassium hydroxide, adding phosphate buffer solution-acetonitrile, dissolving residues through ultrasonic treatment, filtering to obtain filtrate which is used as a testing solution; and (5) adopting HPLC or HPLC-MS to separate and measure the testing solution prepared by the step (4).

Description

technical field

[0001] The invention relates to the field of analytical chemistry, in particular to a method for separating and measuring acetylcysteine ​​enantiomers. Background technique

[0002] Pre-column derivatization is a method to improve detection sensitivity. For most compounds with no UV absorption or weak UV absorption, with appropriate derivatization methods, accurate determination including content and enantiomers can be carried out.

[0003] Acetylcysteine ​​is an amino acid drug that acts as an expectorant. Its chemical name is N-acetyl-L-cysteine ​​and its molecular formula is C 5 h 9 NO 3 S, the chemical name of its enantiomer is N-acetyl-D-cysteine. There is no relevant report on the determination of acetylcysteine ​​isomers in the existing technology, only the literature HPLC Separation of DL-Amino Acide Derivatized with N 2 -(5-Fluoro-2, 4-Dinitrophhenyl)-LAmino Acid Amides (Journal of Chromatography B 674 (1995) 143-148) and the method of Chinese p...

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