Duka Ivan, Gerić Marko, Gajski Goran, Friščić Maja, Maleš Željan, Domijan Ana-Marija, Turčić Petra
Department of Pharmaceutical Botany, Faculty of Pharmacy and Biochemistry, University of Zagreb, Zagreb, Croatia.
Mutagenesis Unit, Institute for Medical Research and Occupational Health, Zagreb, Croatia.
J Environ Sci Health A Tox Hazard Subst Environ Eng. 2020;55(3):275-280. doi: 10.1080/10934529.2019.1687236. Epub 2019 Nov 5.
An adequate level of low molecular weight thiols (LMW-SH, especially glutathione (GSH)) protects cellular macromolecules against toxic agents, and is used as a sensitive biomarker of exposure to toxic compounds. During sample collection, storage and preparation, non-enzymatic and enzymatic oxidation of LMW-SH can occur leading to analytical inaccuracy. The aim of this study was to optimize a fast and reliable screening method for the determination of LMW-SH, mainly GSH, in blood and plasma samples as well as to investigate the impact of storage conditions on the LMW-SH stability. Based on our results, the described spectrophotometric method allows fast and reliable determination of LMW-SH in blood and plasma samples. Results on incubation of samples at 37 °C imply that synthesis of LMW-SH (probably GSH) as well as dynamic interexchange among various thiols forms can be induced in blood cells in conditions. Importantly, the level of LMW-SH in blood and plasma stored at -20 °C was constant, indicating that they can be stored at -20 °C for at least 30 days. Therefore, the method is suitable for assessment of LMW-SH in long-term human biomonitoring as well as environmental field studies, especially those involving a large number of samples such as epidemiological studies.
足够水平的低分子量硫醇(LMW-SH,尤其是谷胱甘肽(GSH))可保护细胞大分子免受有毒物质侵害,并用作接触有毒化合物的敏感生物标志物。在样品采集、储存和制备过程中,LMW-SH可能会发生非酶促和酶促氧化,导致分析不准确。本研究的目的是优化一种快速可靠的筛选方法,用于测定血液和血浆样品中的LMW-SH,主要是GSH,并研究储存条件对LMW-SH稳定性的影响。根据我们的结果,所描述的分光光度法可快速可靠地测定血液和血浆样品中的LMW-SH。在37°C孵育样品的结果表明,在某些条件下血细胞中可诱导LMW-SH(可能是GSH)的合成以及各种硫醇形式之间的动态相互交换。重要的是,储存在-20°C的血液和血浆中LMW-SH的水平是恒定的,这表明它们可以在-20°C储存至少30天。因此,该方法适用于长期人体生物监测以及环境领域研究中LMW-SH的评估,尤其是那些涉及大量样品的研究,如流行病学研究。