Bouligand Jérôme, Deroussent Alain, Paci Angelo, Morizet Jackie, Vassal Gilles
UPRES EA3535 Pharmacology and New Cancer Treatments, Institute Gustave Roussy and Paris XI University, Villejuif, France.
J Chromatogr B Analyt Technol Biomed Life Sci. 2006 Feb 17;832(1):67-74. doi: 10.1016/j.jchromb.2005.12.037. Epub 2006 Jan 24.
A liquid chromatography/tandem mass spectrometry assay of glutathione (GSH), glutathione disulfide (GSSG) and of precursors (gamma-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.
建立了一种用于研究小鼠肝脏中谷胱甘肽（GSH）、谷胱甘肽二硫化物（GSSG）及其前体（γ-谷氨酰半胱氨酸、半胱氨酰甘氨酸、半胱氨酸、胱氨酸、同型半胱氨酸和同型胱氨酸）的液相色谱/串联质谱分析法。经碘乙酸衍生化后，使用反相梯度高效液相色谱法对分析物进行分析，并采用多反应监测进行检测。对含硫醇前体在100 - 10,000 ng/mL浓度范围内进行线性校准，对GSH和GSSG的线性校准范围扩展至100,000 ng/mL。该方法对每种化合物进行了验证，日间准确度低于11.9%，精密度低于15%。该方法显示每种含硫醇化合物和GSSG的定量下限为100 ng/mL，其他二硫化物的定量下限为200 ng/mL。
