Study of disulfide reduction and alkyl chloroformate derivatization of plasma sulfur amino acids using gas chromatography-mass spectrometry

Analytical and Bioanalytical Chemistry
Zdeněk SvageraPetr Hušek

Abstract

Four disulfide-reducing agents, dithiothreitol (DTT), 2,3-dimercaptopropanesulfonate (DMPS), and the newly tested 2-mercaptoethanesulfonate (MESNA) and Tris(hydroxypropyl)phosphine (THP), were investigated in detail for release of sulfur amino acids in human plasma. After protein precipitation with trichloroacetic acid (TCA), the plasma supernatant was treated with methyl, ethyl, or propyl chloroformate via the well-proven derivatization-extraction technique and the products were subjected to gas chromatographic-mass spectrometric (GC-MS) analysis. All the tested agents proved to be rapid and effective reducing agents for the assay of plasma thiols. When compared with DTT, the novel reducing agents DMPS, MESNA, and THP provided much cleaner extracts and improved analytical performance. Quantification of homocysteine, cysteine, and methionine was performed using their deuterated analogues, whereas other analytes were quantified by means of 4-chlorophenylalanine. Precise and reliable assay of all examined analytes was achieved, irrespective of the chloroformate reagent used. Average relative standard deviations at each analyte level were ≤6%, quantification limits were 0.1-0.2 μmol L(-1), recoveries were 94-121%, and linearity wa...Continue Reading

References

Aug 7, 2001·Journal of Chromatography. B, Biomedical Sciences and Applications·Y ShinoharaT Hashimoto
Oct 11, 2001·Clinical Chemistry and Laboratory Medicine : CCLM·M NauckH Wieland
Oct 27, 2001·Journal of Chromatography. B, Biomedical Sciences and Applications·J B LaurensW J Vermaak
May 14, 2003·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Petr HusekPetr Simek
Sep 6, 2005·Clinica Chimica Acta; International Journal of Clinical Chemistry·Katharina SchröcksnadelDietmar Fuchs
Oct 30, 2007·Clinical Chemistry and Laboratory Medicine : CCLM·Per Magne UelandSteinar Hustad
May 30, 2008·Clinical Chemistry·Per Magne Ueland
Apr 10, 2009·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Toshimasa Toyo'oka
Jun 2, 2009·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Mahroukh RafiiPaul B Pencharz
Jun 12, 2009·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·M Elizabeth McMenaminThomas D Nolin
Feb 24, 2012·Comparative Biochemistry and Physiology. Part A, Molecular & Integrative Physiology·P JedličkaH Zahradníčková

❮ Previous
Next ❯

Citations

Aug 15, 2014·Molecular Microbiology·Gustavo A AfanadorSean T Prigge
Oct 21, 2014·Evolution; International Journal of Organic Evolution·Ismael GalvánCarlos Alonso-Alvarez
May 27, 2017·The Journal of Experimental Biology·Ismael Galván, Carlos Alonso-Alvarez

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.