Simultaneous Determination of Human Serum Albumin and Low-Molecular-Weight Thiols after Derivatization with Monobromobimane.
Simultaneous Determination of Human Serum Albumin and Low-Molecular-Weight Thiols after Derivatization with Monobromobimane.
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DOI：
10.3390/molecules26113321
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发表时间：
2021-06-01
期刊：
影响因子：
--
通讯作者：
Chwatko G
中科院分区：
文献类型：
--
作者：
Kurpet K;Głowacki R;Chwatko G
生物硫醇是非常强大的抗氧化剂，可以保护细胞免受氧化应激的影响。它们也被认为是相关的疾病生物标志物，特别是心血管疾病的危险因素。本文介绍了一种同时测定血浆中人血清白蛋白和低分子硫醇的新方法。该方法采用巯基特异性荧光标记试剂单溴马烷对分析物进行柱前衍生化，然后采用荧光检测（激发波长为378nm，发射波长为492nm）的反相高效液相色谱分离和定量。在衍生化步骤之前，氧化的硫醇通过硼氢化钠的还原裂解转化为它们的还原形式。总硫醇在以下范围内呈线性关系：人血清白蛋白为1.76 ~ 30.0 mg mL - 1， α-硫辛酸为0.29 ~ 5.0 nmol mL - 1，谷胱甘肽为1.16 ~ 35 nmol mL - 1，半胱氨酸为9.83 ~ 450.0 nmol mL - 1，同型半胱氨酸为0.55 ~ 40.0 nmol mL - 1， n -乙酰-l -半胱氨酸为0.34 ~ 50.0 nmol mL - 1，半胱氨酸为1.45 ~ 45.0 nmol mL - 1。所有分析物的回收率为85.16 ~ 119.48%。该方法灵敏度高，重复性好，在总硫醇的测定范围内呈线性。所设计的程序可应用于血浆样品，以监测各种病理生理状态下的生化过程。
Biothiols are extremely powerful antioxidants that protect cells against the effects of oxidative stress. They are also considered relevant disease biomarkers, specifically risk factors for cardiovascular disease. In this paper, a new procedure for the simultaneous determination of human serum albumin and low-molecular-weight thiols in plasma is described. The method is based on the pre-column derivatization of analytes with a thiol-specific fluorescence labeling reagent, monobromobimane, followed by separation and quantification through reversed-phase high-performance liquid chromatography with fluorescence detection (excitation, 378 nm; emission, 492 nm). Prior to the derivatization step, the oxidized thiols are converted to their reduced forms by reductive cleavage with sodium borohydride. Linearity in the detector response for total thiols was observed in the following ranges: 1.76–30.0 mg mL−1 for human serum albumin, 0.29–5.0 nmol mL−1 for α-lipoic acid, 1.16–35 nmol mL−1 for glutathione, 9.83–450.0 nmol mL−1 for cysteine, 0.55–40.0 nmol mL−1 for homocysteine, 0.34–50.0 nmol mL−1 for N-acetyl-L-cysteine, and 1.45–45.0 nmol mL−1 for cysteinylglycine. Recovery values of 85.16–119.48% were recorded for all the analytes. The developed method is sensitive, repeatable, and linear within the expected ranges of total thiols. The devised procedure can be applied to plasma samples to monitor biochemical processes in various pathophysiological states.
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DOI：
10.1080/10826070903249666
发表时间：
2009-01-01
影响因子：
1.3
作者：
Glowacki, R.;Bald, E.
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影响因子：
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DOI：
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发表时间：
2012-12-12
影响因子：
3
作者：
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影响因子：
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