Glutathione measurement in human plasma. Evaluation of sample collection, storage and derivatization conditions for analysis of dansyl derivatives by HPLC.
Literature values for human plasma GSH vary over 10-fold despite the use of apparently valid analytical procedures for GSH measurement. The purpose of this study was to develop a procedure to minimize error in sample collection, processing and storage that could contribute to such differences. HPLC with fluorescence detection of dansyl derivatives was used for quantification. The results show that collection of blood with a butterfly needle and syringe reduces overestimation due to limited hemolysis and that use of a preservation solution designed to inhibit autooxidation and enzymatic degradation allows quantitative recovery of both GSH and GSSG. Stability tests showed that non-derivatized samples were stable for at least 2 months at - 80 degrees while dansyl derivatives were stable in the dark at 0-4 degrees for 12 months. Results from 59 healthy individuals (20-43 years) provided a mean (+/-1 SD) GSH value of 2.09+/-1.14 micromolar.
Department of Biochemistry, Emory University School of Medicine, Atlanta, GA 30322, USA. dpjones@emory.edu
D P
DP
Jones
J L
JL
Carlson
P S
PS
Samiec
P
P
Sternberg
V C
VC
Mody
R L
RL
Reed
L A
LA
Brown
D001800:Blood Specimen Collection / Q000295:instrumentation / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D003619:Dansyl Compounds / Q000737:chemistry; D005978:Glutathione / Q000097:blood* / Q000737:chemistry; D019803:Glutathione Disulfide / Q000097:blood; D006801:Humans; D007202:Indicators and Reagents / Q000737:chemistry; D013050:Spectrometry, Fluorescence
D003619:Dansyl Compounds; D007202:Indicators and Reagents; D005978:Glutathione; D019803:Glutathione Disulfide