Stable isotope dilution mass spectrometry quantification of hydrogen sulfide and thiols in biological matrices

被引：21

作者：

Malaeb, Hind ^{[1
,2
,3
]}

Choucair, Ibrahim ^{[1
,3
]}

Wang, Zeneng ^{[1
,3
]}

Li, Xinmin S. ^{[1
,3
]}

Li, Lin ^{[1
,3
]}

Boyd, W. Christopher ^{[2
]}

Hine, Christophe ^{[1
,3
]}

Tang, W. H. Wilson ^{[1
,3
,4
]}

Gogonea, Valentin ^{[1
,2
,3
,6
]}

Hazen, Stanley L. ^{[1
,2
,3
,4
,5
]}

机构：

[1] Cleveland Clin, Lerner Res Inst, Dept Cardiovasc & Metab Sci, Cleveland, OH USA

[2] Cleveland State Univ, Dept Chem, Cleveland, OH USA

[3] Cleveland Clin, Ctr Microbiome & Human Hlth, Cleveland, OH USA

[4] Cleveland Clin, Dept Cardiovasc Med, Heart Vasc & Thorac Inst, Cleveland, OH USA

[5] Cleveland Clin, Lerner Res Inst, Dept Cardiovasc & Metab Sci, 9500 Euclid Ave,NC-10, Cleveland, OH 44195 USA

[6] Cleveland State Univ, Dept Chem, 2121 Euclid Ave, Cleveland, OH 44115 USA

来源：

REDOX BIOLOGY | 2022年 / 55卷

基金：

美国国家卫生研究院;

关键词：

Hydrogen sulfide; Gut microbiota; Aging; Liquid chromatography tandem mass; spectrometry; Plasma thiol; Cysteine; Homocysteine; Glutathione; Cysteinylglycine; gamma-glutamylcysteine; Microbiome; CAPACITY; CYSTEINE; KIDNEY; STATE; H2S;

D O I：

10.1016/j.redox.2022.102401

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Background: Hydrogen sulfide (H2S), a gaseous signaling molecule that impacts multiple physiological processes including aging, is produced via select mammalian enzymes and enteric sulfur-reducing bacteria. H2S research is limited by the lack of an accurate internal standard-containing assay for its quantitation in biological matrices. Methods: After synthesizing [S-34]H2S and developing sample preparation protocols that avoid sulfide contami-nation with the addition of thiol-containing standards or reducing reagents, we developed a stable isotope-dilution high performance liquid chromatography tandem-mass spectrometry (LC-MS/MS) method for the simultaneous quantification of Total H2S and other abundant thiols (cysteine, homocysteine, glutathione, glu-tamylcysteine, cysteinylglycine) in biological matrices, conducted a 20-day analytical validation/normal range study, and then both analyzed circulating Total H2S and thiols in plasma from 400 subjects, and within 20 volunteers before and after antibiotic-induced suppression of gut microbiota. Results: Using the new assay, all analytes showed minimal interference, no carryover, and excellent intra-and inter-day reproducibility (<= 7.6%, and <= 12.7%, respectively), linearity (r(2) > 0.997), recovery (90.9%-110%) and stability (90.0%-100.5%). Only circulating Total H2S levels showed significant age-associated reductions in both males and females (p < 0.001), and a marked reduction following gut microbiota suppression (mean 33.8 +/- 17.7%, p < 0.001), with large variations in gut microbiota contribution among subjects (range 6.0-66.7% reduction with antibiotics). Conclusions: A stable-isotope-dilution LC-MS/MS method is presented for the simultaneous quantification of Total H2S and multiple thiols in biological matrices. We then use this assay panel to show a striking age-related decline and gut microbiota contribution to circulating Total H2S levels in humans.

引用

页数：10