Determination of Total Glutathione in Dried Blood Spot Samples Using a High-Performance Liquid Chromatography

A method is described for the determination of total glutathione (TGSH) in dried blood spot (DBS) samples using high-performance liquid chromatography with fluorescence detection. Whole blood and DBS samples were obtained from a group of blood donors. After GSH reduction with dithiothreitol and prot...

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Bibliographic Details
Published in:Journal of chromatographic science 2015-07, Vol.53 (6), p.879-885
Main Authors: Kanďár, Roman, Štramová, Xenie, Drábková, Petra, Brandtnerová, Martina
Format: Article
Language:English
Subjects:
Adolescent
Adult
Chromatography, High Pressure Liquid - methods
Dried Blood Spot Testing - methods
Female
Glutathione - blood
Glutathione - chemistry
Humans
Limit of Detection
Linear Models
Male
Methanol
Middle Aged
Reproducibility of Results
Young Adult
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Summary:A method is described for the determination of total glutathione (TGSH) in dried blood spot (DBS) samples using high-performance liquid chromatography with fluorescence detection. Whole blood and DBS samples were obtained from a group of blood donors. After GSH reduction with dithiothreitol and protein precipitation with ethanol, the samples were derivatized with naphthalene-2,3-dicarboxaldehyde to form a very stable, highly fluorescent derivative. For the separation, a reversed phase HPLC method was used. The mixture of ethanol and deionized water (8 : 92, v/v) was used as a mobile phase. The analytical performance of this method was satisfactory: the intra- and interassay coefficients of variation were below 10%. Quantitative recoveries from spiked DBS samples were between 98.3 and 103.6%. The presented method is inexpensive and suitable for clinical testing purposes.
ISSN:0021-9665
1945-239X
DOI:10.1093/chromsci/bmu135