title: Mass spectrometry based detection of glutathione with sensitivity for single-cell analysis.
abstract: BACKGROUND
  Glutathione (GSH) is a very important molecule that participates in various physiologically important events. Depletion of cellular GSH has been suggested as a biomarker of early stage of cell injury. Assessment of GSH with high selectivity and sensitivity is in demand in many fields of life sciences.
  
  
  METHODS
  Cell lysates were deproteinated by 5-sulfosalicylic acid. Glutathione was derivatized with monobromobimane to form a GSH conjugate. S-Hexylglutathione was employed as internal standard. The resulting samples were analyzed using a high-performance liquid chromatography/electrospray ionization tandem mass spectrometry (HPLC/ESI-MS/MS) system.
  
  
  RESULTS
  The limit of detection was as low as 500 amol with high selectivity. The approach was sensitive enough to detect GSH content in a single human erythrocyte. The technique is the most sensitive approach among the reported MS-based GSH assays.
  
  
  CONCLUSIONS
  A monobromobimane derivatization and mass spectrometry based method was developed for the assessment of GSH. The derivatized GSH was chemically stable against autooxidation. With the great selectivity and sensitivity, the approach is anticipated to serve as a routine method for assessment of GSH and possibly other important endogenous low molecular weight thiols.
authors.affiliation: School of Traditional Chinese Medicine, Shenyang Pharmaceutical University, Shenyang, Liaoning, P.R. China.
authors.forename: Jia
authors.initials: J
authors.lastname: Yu
authors.affiliation: School of Traditional Chinese Medicine, Shenyang Pharmaceutical University, Shenyang, Liaoning, P.R. China.
authors.forename: Chunyan
authors.initials: C
authors.lastname: Li
authors.affiliation: Center for Developmental Therapeutics, Seattle Children's Research Institute; Division of Hepatology and Gastroenterology, Department of Pediatrics, University of Washington School of Medicine, Seattle, WA, USA.
authors.forename: Shuijie
authors.initials: S
authors.lastname: Shen
authors.affiliation: School of Traditional Chinese Medicine, Shenyang Pharmaceutical University, Shenyang, Liaoning, P.R. China.
authors.forename: Xiaoqiu
authors.initials: X
authors.lastname: Liu
authors.affiliation: School of Traditional Chinese Medicine, Shenyang Pharmaceutical University, Shenyang, Liaoning, P.R. China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Peng
authors.affiliation: Center for Developmental Therapeutics, Seattle Children's Research Institute; Division of Hepatology and Gastroenterology, Department of Pediatrics, University of Washington School of Medicine, Seattle, WA, USA.|Key Laboratory of Structure-Based Drug Design & Discovery of Ministry of Education, Shenyang Pharmaceutical University, Shenyang, Liaoning, P.R. China.
authors.forename: Jiang
authors.initials: J
authors.lastname: Zheng
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005978:Glutathione / Q000032:analysis*; D006801:Humans; D057230:Limit of Detection; D051381:Rats; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D059010:Single-Cell Analysis / Q000379:methods*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D005978:Glutathione