+ Site Statistics
References:
52,654,530
Abstracts:
29,560,856
PMIDs:
28,072,755
+ Search Articles
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ PDF Full Text
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn

+ Translate
+ Recently Requested

The determination of glutathione, cyst(e)ine, and other thiols and disulfides in biological samples using high-performance liquid chromatography with dual electrochemical detection



The determination of glutathione, cyst(e)ine, and other thiols and disulfides in biological samples using high-performance liquid chromatography with dual electrochemical detection



Analytical Biochemistry 163(1): 9-15



A determination of glutathione, cysteine, and their disulfides using HPLC and dual electrochemical detection (HPLC-DEC) was described previously but was not validated in biological tissues for these and other important thiols and disulfides (SH/SS). Thus, our objectives were to develop this method to quantify simultaneously reduced and oxidized glutathione, cysteine, cystine, and other SH/SS in various tissues, including human blood and plasma, rat liver and hippocampus, mosquito, and spinach leaf. Optimal conditions were determined for sample processing and analysis using metaphosphoric acid and HPLC-DEC. Authentic standards of 10 common SH/SS compounds were resolved and eluted within 15 min, and all standard curves were linear from 5 to 1600 pmol. Validation was based on the following: First, tissue sample sizes were proportional to peak areas over an eightfold range. Second, recovery of SH/SS added to samples before processing was 96-101%. Finally, the results were equivalent and correlated highly with values for total SH by 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB) assay (r2 = 0.996) and for total glutathione by DTNB-GSSG reductase assay (r2 = 0.998). The life span of the Au/Hg electrode was limited to 200-500 samples based on the lineal range of standard curves. On the basis of these results, we believe that this method will fill a long-time need for the simultaneous determination of SH/SS in biological tissues.

(PDF emailed within 0-6 h: $19.90)

Accession: 041605588

Download citation: RISBibTeXText

PMID: 3619033

DOI: 10.1016/0003-2697(87)90085-6



Related references

The determination of glutathione cysteine and other thiols and disulfides in biological samples using high performance liquid chromatography and dual electrochemical detection. Federation Proceedings: 1733, 1986

Determination of thiols and disulfides using high-performance liquid chromatography with electrochemical detection. Journal Of Chromatography B Biomedical Applications. 672(1): 73-80, 1995

Automated high performance liquid chromatography with on-line reduction of disulfides and chemiluminescence detection for determination of thiols and disulfides in biological fluids. Analytica Chimica Acta 768: 96-101, 2013

Measurement of biological thiols and disulfides by high-performance liquid chromatography and electrochemical detection of silver mercaptide formation. Analytical Biochemistry 197(1): 19-24, 1991

Simultaneous determination of thiols and corresponding disulfides in mixtures by high performance liquid chromatography with polarographic detection using a dual tast analyzer. Journal of Chromatography 455: 336-343, 1988

Glutathione and cyst(e)ine profiles of vegetables using high performance liquid chromatography with dual electrochemical detection. Journal of Food Composition and Analysis 10(2): 90-101, 1997

Determination of glutathione in biological material by high-performance liquid chromatography with electrochemical detection. Analytica Chimica Acta 196: 251-254, 1987

Determination of glutathione in biological tissues by high-performance liquid chromatography with electrochemical detection. Journal of Pharmaceutical and Biomedical Analysis 7(5): 643-647, 1989

Determination of oxidized and reduced glutathione in pharmaceuticals by reversed-phase high-performance liquid chromatography with dual electrochemical detection. Journal of Chromatography A 846(1-2): 59-64, June 18, 1999

High-performance liquid chromatography using electrochemical detection for the determination of prazosin in biological samples. Journal Of Chromatography B Biomedical Applications. 670(1): 177-182, 1995

Determination of prilocaine in human plasma samples using high-performance liquid chromatography with dual-electrode electrochemical detection. Journal of Chromatography 526(1): 215-222, 1990

Determination of glutathione in biological samples by high performance liquid chromatography with fluorescence detection. Biomedical Chromatography 8(6): 306-308, 1994

Speciation and determination of thiols in biological samples using high performance liquid chromatography-inductively coupled plasma-mass spectrometry and high performance liquid chromatography-Orbitrap MS. Analytica Chimica Acta 680(1-2): 41-47, 2011

Determination of the enantiomeric composition of salsolinol in biological samples by high-performance liquid chromatography with electrochemical detection. Journal of Chromatography 495: 205-214, 1989

Determination of clenbuterol in biological samples by high performance liquid chromatography with in series ultraviolet and electrochemical detection. Wei Sheng Yan Jiu 33(5): 623-4, 628, 2004