TY - JOUR
T1 - Liquid chromatography-tandem mass spectrometry assay of reduced and oxidized glutathione and main precursors in mice liver
AU - Bouligand, Jérôme
AU - Deroussent, Alain
AU - Paci, Angelo
AU - Morizet, Jackie
AU - Vassal, Gilles
N1 - Funding Information:
This work was supported by a grant from the ARC (Association de la Recherche sur le Cancer). The Institute Gustave Roussy, the C.N.R.S. (Centre National de la Recherche Scientifique), the Ligue contre le Cancer and the ARC are gratefully acknowledged for the acquisition of the tandem mass spectrometer. We thank Patrice Ardouin and the staff of the animal unit at the Institute Gustave Roussy for their care of the animals. We thank Jean-Pierre Levillain for peptide derivatization methods.
PY - 2006/2/17
Y1 - 2006/2/17
N2 - A liquid chromatography/tandem mass spectrometry assay of glutathione (GSH), glutathione disulfide (GSSG) and of precursors (γ-glutamyl- cysteine, cysteinyl-glycine, cysteine, cystine, homocysteine and homocystine) was developed to study glutathione synthesis in mice liver. After iodoacetic acid derivatization, the analytes were analyzed using reversed-phase gradient HPLC and detected using multiple reaction monitoring. Linear calibrations were performed over the concentrations range of 100-10,000 ng/mL for the thiol-containing precursors and extended up to 100,000 ng/mL for GSH and GSSG. The method was validated for each compound with inter-day accuracy below 11.9% and with precision below 15%. The method showed low limits of quantitation of 100 ng/mL for each thiol-containing compound and GSSG and of 200 ng/mL for other disulfides.
AB - A liquid chromatography/tandem mass spectrometry assay of glutathione (GSH), glutathione disulfide (GSSG) and of precursors (γ-glutamyl- cysteine, cysteinyl-glycine, cysteine, cystine, homocysteine and homocystine) was developed to study glutathione synthesis in mice liver. After iodoacetic acid derivatization, the analytes were analyzed using reversed-phase gradient HPLC and detected using multiple reaction monitoring. Linear calibrations were performed over the concentrations range of 100-10,000 ng/mL for the thiol-containing precursors and extended up to 100,000 ng/mL for GSH and GSSG. The method was validated for each compound with inter-day accuracy below 11.9% and with precision below 15%. The method showed low limits of quantitation of 100 ng/mL for each thiol-containing compound and GSSG and of 200 ng/mL for other disulfides.
KW - Disulfide glutathione
KW - Glutathione
KW - Glutathione precursors
KW - HPLC-MS/MS
KW - Liver
KW - Mice
UR - http://www.scopus.com/inward/record.url?scp=32244432846&partnerID=8YFLogxK
U2 - 10.1016/j.jchromb.2005.12.037
DO - 10.1016/j.jchromb.2005.12.037
M3 - Article
C2 - 16434243
AN - SCOPUS:32244432846
SN - 1570-0232
VL - 832
SP - 67
EP - 74
JO - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
JF - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
IS - 1
ER -