Improved method for the determination of N-acetylcysteine in human plasma by high-performance liquid chromatography.
An improved method for the determination of N-acetylcysteine by paired-ion reversed-phase high-performance liquid chromatography has been developed. Following incubation with dithiothreitol to release bound N-acetylcysteine, free N-acetylcysteine and the internal standard N-acetylpenicillamine were derivatised with 2,4-dinitro-1-fluorobenzene. The samples were then deproteinised by ultrafiltration. The dinitrophenyl derivatives were extracted from acidified ultrafiltrate into diethyl ether and purified by using a back-extraction step. They were then separated from naturally occurring plasma components and reagent impurities by high-performance liquid chromatography, utilising an Ultrasphere ODS (5 microns) reversed-phase column and detection at 360 nm.
D000111:Acetylcysteine / Q000097:blood*; D055598:Chemical Phenomena; D002621:Chemistry; D002851:Chromatography, High Pressure Liquid; D004139:Dinitrofluorobenzene; D006801:Humans; D010396:Penicillamine / Q000031:analogs & derivatives / Q000097:blood
C010005:N-acetylpenicillamine; D004139:Dinitrofluorobenzene; D010396:Penicillamine; D000111:Acetylcysteine