Method for separating and measuring acetylcysteine enantiomers

An acetylcysteine ​​and enantiomer technology, applied in the field of analytical chemistry, can solve problems such as unavailability of normal phase conditions, limited types of mobile phases, and difficulty in detection sensitivity, achieving stable product and fast reaction speed. , the effect of mild reaction conditions

Active Publication Date: 2011-02-09
湖北新生源生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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...

Method used

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  • Method for separating and measuring acetylcysteine enantiomers
  • Method for separating and measuring acetylcysteine enantiomers
  • Method for separating and measuring acetylcysteine enantiomers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] (1) Instruments and conditions

[0029] Diane UltiMate 3000 high performance liquid chromatography system and workstation in the United States, automatic sample injection, the chromatographic column uses octadecylsilane bonded silica gel column; UV detection wavelength: 333nm. Mobile phase: Use 0.02mol / L ammonium acetate solution (adjust the pH value to 6.8 with phosphoric acid or ammonia water) as mobile phase A, acetonitrile as mobile phase B, and perform linear gradient elution in the following table

[0030] time (minutes)

Mobile phase A (%)

Mobile phase B (%)

0

80

20

18

80

20

19

50

50

25

50

50

26

80

20

36

80

20

[0031] (2) Experimental steps

[0032] Take 11mg of D-acetylcysteine, add water and dissolve it into 25ml solution, add L-acetylcysteine ​​82mg and dissolve it to make a mixed solution, take 1ml of the mixed solution and put it in a 10ml measuring bottle, add 2...

Embodiment 2

[0034] (1) Instruments and conditions

[0035] Diane UltiMate 3000 high performance liquid chromatography system and workstation in the United States, automatic sample injection, the chromatographic column uses octadecylsilane bonded silica gel column; UV detection wavelength: 333nm. Mobile phase: use 0.03mol / L ammonium acetate solution (adjust the pH value to 5.0 with phosphoric acid or ammonia water) as mobile phase A, acetonitrile as mobile phase B, and perform linear gradient elution in the following table

[0036] time (minutes)

Mobile phase A (%)

Mobile phase B (%)

0

80

20

18

80

20

19

50

50

25

50

50

26

80

20

36

80

20

[0037] (2) Experimental steps

[0038] Take 82mg of acetylcysteine ​​and put it in a 10ml measuring bottle, add 2ml of 1mol / L sulfuric acid solution, shake to dissolve, and neutralize with 1mol / L potassium hydroxide solution to pH 6.0, add water to dilute to th...

Embodiment 3

[0040] (1) Instruments and conditions

[0041] Diane UltiMate 3000 high performance liquid chromatography system and workstation in the United States, automatic sample injection, the chromatographic column uses octadecylsilane bonded silica gel column; UV detection wavelength: 333nm. Mobile phase: use 0.04mol / L ammonium acetate solution (adjust the pH value to 7.0 with phosphoric acid or ammonia water) as mobile phase A, acetonitrile as mobile phase B, and perform linear gradient elution in the following table

[0042] time (minutes)

Mobile phase A (%)

Mobile phase B (%)

0

80

20

18

80

20

19

50

50

25

50

50

26

80

20

36

80

20

[0043] (2) Experimental steps

[0044] Take 82mg of acetylcysteine ​​granules, put them in a 10ml measuring bottle, add 2ml of 1mol / L formic acid solution, shake to dissolve, neutralize with 1mol / L ammonia solution to pH 7.0, add water to dilute to the mark, sh...

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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...

Claims

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Application Information

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IPC IPC(8): G01N30/14G01N30/02
Inventor 王先兵程文莹杨娟吕启超郭刚
Owner 湖北新生源生物工程有限公司
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