Validation of a simplified procedure for convenient and rapid quantification of reduced and oxidized glutathione in human plasma by liquid chromatography tandem mass spectrometry analysis.
Validation of a simplified procedure for convenient and rapid quantification of reduced and oxidized glutathione in human plasma by liquid chromatography tandem mass spectrometry analysis.
复制标题
DOI：
10.1002/bmc.4854
复制
发表时间：
2020-09
期刊：
影响因子：
--
通讯作者：
Levy MA
中科院分区：
文献类型：
--
作者：
Enomoto AC;Schneider E;McKinnon T;Goldfine H;Levy MA
内源性谷胱甘肽（GSH）和谷胱甘肽二硫化物（GSSG）状态对氧化条件高度敏感，并作为体内氧化还原状态的替代指标具有广泛的应用。已建立的全血中GSH和GSSG定量方法在人血浆中显示出有限的实用性，其中GSH和GSSG水平比全血中观察到的水平低约3-4个数量级。本研究提供了简化的样品处理和分析LC-MS/MS方法，显示了测量人血浆样品中GSH和GSSG浓度所需的灵敏度和准确度，在5倍稀释以抑制基质干扰后，其范围为200 - 500 nm（GSH）和5-30 nm（GSSG）。本文报告的方法的实用性通过试验性能和验证参数得到证明，这些参数表明灵敏度良好[定量下限为4.99 nm（GSH）和3.65 nm（GSSG）]，试验精密度较高（GSH和GSSG的试验内CV分别为3.6%和1.9%，试验间CV分别为7.0%和2.8%）。这些方法还显示分析物加标血浆样品的回收率异常（GSH为98.0 ± 7.64%，GSSG为98.5 ± 12.7%）。GSH和GSSG在−80°C下的稳定性分别为55周和46周，平均CV分别<15和<10%。
Endogenous glutathione (GSH) and glutathione disulfide (GSSG) status is highly sensitive to oxidative conditions and have broad application as a surrogate indicator of redox status in vivo. Established methods for GSH and GSSG quantification in whole blood display limited utility in human plasma, where GSH and GSSG levels are ~3–4 orders of magnitude below those observed in whole blood. This study presents simplified sample processing and analytical LC–MS/MS approaches exhibiting the sensitivity and accuracy required to measure GSH and GSSG concentrations in human plasma samples, which after 5‐fold dilution to suppress matrix interferences range from 200 to 500 nm (GSH) and 5–30 nm (GSSG). The utility of the methods reported herein is demonstrated by assay performance and validation parameters which indicate good sensitivity [lower limits of quantitation of 4.99 nm (GSH) and 3.65 nm (GSSG), and high assay precision (intra‐assay CVs 3.6 and 1.9%, and inter‐assay CVs of 7.0 and 2.8% for GSH and GSSG, respectively). These methods also exhibited exceptional recovery of analyte‐spiked plasma samples (98.0 ± 7.64% for GSH and 98.5 ± 12.7% for GSSG). Good sample stability at −80°C was evident for GSH for up to 55 weeks and GSSG for up to 46 weeks, with average CVs <15 and <10%, respectively.
登录
查看更多内容
影响因子：
3.3
作者：
ROUM, JH;BUHL, R;CRYSTAL, RG
通讯作者：
CRYSTAL, RG
DOI：
10.1016/0378-4347(95)00245-e
发表时间：
1995-11-03
期刊：
JOURNAL OF CHROMATOGRAPHY B-BIOMEDICAL APPLICATIONS
影响因子：
--
作者：
SMITH, NC;DUNNETT, M;MILLS, PC
通讯作者：
MILLS, PC
影响因子：
14.8
作者：
Giustarini, Daniela;Dalle-Donne, Isabella;Rossi, Ranieri
通讯作者：
Rossi, Ranieri
影响因子：
7.4
作者：
Carroll D;Howard D;Zhu H;Paumi CM;Vore M;Bondada S;Liang Y;Wang C;St Clair DK
通讯作者：
St Clair DK
DOI：
10.1016/j.jpba.2018.10.053
发表时间：
2019-02-05
影响因子：
3.4
作者：
Forgacsova, Andrea;Galba, Jaroslav;Kovac, Andrej
通讯作者：
Kovac, Andrej