High performance liquid chromatographic assay for the quantitation of total glutathione in plasma.
A simple and widely used homocysteine HPLC procedure was applied for the HPLC identification and quantitation of glutathione in plasma. The method, which utilizes SBDF as a derivatizing agent utilizes only 50 microl of sample volume. Linear quantitative response curve was generated for glutathione over a concentration range of 0.3125-62.50 micromol/l. Linear regression analysis of the standard curve exhibited correlation coefficient of 0.999. Limit of detection (LOD) and limit of quantitation (LOQ) values were 5.0 and 15 pmol, respectively. Glutathione recovery using this method was nearly complete (above 96%). Intra-assay and inter-assay precision studies reflected a high level of reliability and reproducibility of the method. The applicability of the method for the quantitation of glutathione was demonstrated successfully using human and rat plasma samples.
Abukhalaf
Imad K
IK
Department of Pharmacology and Toxicology, Morehouse School of Medicine, Atlanta, GA 30310, USA. abukhai@msm.edu
Silvestrov
Natalia A
NA
Menter
Julian M
JM
von Deutsch
Daniel A
DA
Bayorh
Mohamed A
MA
Socci
Robin R
RR
Ganafa
Agaba A
AA
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D005456:Fluorescent Dyes; D005978:Glutathione / Q000097:blood*; D006801:Humans; D007202:Indicators and Reagents; D051381:Rats; D019163:Reducing Agents; D012015:Reference Standards; D015203:Reproducibility of Results; D012996:Solutions
D005456:Fluorescent Dyes; D007202:Indicators and Reagents; D019163:Reducing Agents; D012996:Solutions; D005978:Glutathione