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...

AI Technical Summary

Problems solved by technology

If chiral chromatographic columns are used to separate isomeric impurities, on the one hand, the types of mobile phases are greatly limited, and normal phase conditions are almost unavailable; in addition, the detection sensitivity is not easy to meet the requirements

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