High-performance liquid chromatography method for the determination of protein-glutathione mixed disulfide

Kshama Kumari, Peeyush Khanna, Naseem H. Ansari, Satish K. Srivastava

Research output: Contribution to journalArticle

13 Scopus citations

Abstract

We have developed an efficient and sensitive method for the quantification of picomole quantities of protein-glutathione and protein-cysteine mixed disulfides in ocular lens. Performic acid oxidation releases glutathione and cysteine from the protein-mixed disulfides as glutathione sulfonic acid and cysteic acid, respectively. The sulfonic acids are derivatized with phenylisothiocyanate, and the phenylthiocarbamyl derivatives are separated by HPLC using a reversed-phase ODS column, equilibrated with 0.14 M sodium acetate, containing 0.1% triethylamine and 6% acetonitrile.

Original languageEnglish (US)
Pages (from-to)374-376
Number of pages3
JournalAnalytical Biochemistry
Volume220
Issue number2
DOIs
StatePublished - Aug 1 1994

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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