Journal
JOURNAL OF SEPARATION SCIENCE
Volume 41, Issue 16, Pages -Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/jssc.201800381
Keywords
amino acids; brain homogenate; high-performance liquid chromatography; thiols
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Funding
- University of Lodz [B1711100001599.02]
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We have developed a simple, fast, accurate, and cheap method for the simultaneous determination of total cysteine, homocysteine, glutathione, and N-acetylcysteine in brain homogenates based on the reduction of disulfide bonds by tris(2-carboxyethyl) phosphine, pre-column derivatization of free thiol groups with 2-chloro-1-methylquinolinium tetrafluoroborate followed by ion-pair reversed-phase high-performance liquid chromatography separation with ultraviolet detection. The separation of thiol derivatives was achieved in 10min. Linearity was observed in the range of 10-300, 0.7-10, 2-30, and 3-20mol/L homogenate with a limit of detection of 3.7, 0.2, 0.8, and 1.2mol/L homogenate for cysteine, homocysteine, glutathione, and N-acetylcysteine, respectively. The precision, calculated as relative standard deviation, was in the range of 1.21-4.77, 1.53-14.35, 0.47-1.92, and 1.61-8.95% for cysteine, homocysteine, glutathione, and N-acetylcysteine, respectively. The presented method was successfully applied to the selective determination of total amino thiols in pig brain tissue samples.
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