Wadsworth Center , New York State Department of Health , Empire State Plaza , P.O. Box 509, Albany , New York 12201-0509 , United States.
Department of Environmental Health Sciences, School of Public Health , State University of New York at Albany , Empire State Plaza , P.O. Box 509, Albany , New York 12201-0509 , United States.
Environ Sci Technol. 2018 Jun 5;52(11):6647-6655. doi: 10.1021/acs.est.8b00883. Epub 2018 May 10.
The determination of oxidative stress biomarkers (OSBs) is useful for the assessment of health status and progress of diseases in humans. Whereas previous methods for the determination of OSBs in urine were focused on a single marker, in this study, we present a method for simultaneous determination of biomarkers of oxidative damage to lipids, proteins, and DNA. 2,4-Dinitrophenylhydrazine (DNPH) derivatization followed by solid phase extraction (SPE) and high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) allowed the determination of 8-hydroxy-2'-deoxyguanosine (8-OHdG), o- o'-dityrosine (diY), malondialdehyde (MDA), and four F-isoprostane isomers: 8-iso-prostaglandinF (8-PGF), 11β-prostaglandinF (11-PGF), 15( R)-prostaglandinF (15-PGF), and 8-iso,15( R)-prostaglandinF (8,15-PGF) in urine. Derivatization with DNPH and SPE was optimized to yield greater sensitivity and selectivity for the analysis of target chemicals. The limits of detection of target analytes in urine were below 30 pg mL. The assay intra- and interday variability was below 16% of the relative standard deviation, and the recoveries of target chemicals spiked into synthetic urine were near 100%. The method was applied to the analysis of 21 real urine samples, and the analytes were found at a detection frequency of 85% for 8-PGF and 15-PGF, 71% for 11-PGF, 81% for 8,15-PGF, and 100% for diY, 8-OHdG, and MDA. This method offers simultaneous determination of multiple OSBs of different molecular origin in urine samples selectively with high accuracy and precision.
氧化应激生物标志物（OSB）的测定对于评估人类的健康状况和疾病进展非常有用。虽然以前测定尿液中 OSB 的方法侧重于单个标志物，但在本研究中，我们提出了一种同时测定脂质、蛋白质和 DNA 氧化损伤生物标志物的方法。2,4-二硝基苯肼（DNPH）衍生化后，通过固相萃取（SPE）和高效液相色谱-串联质谱（HPLC-MS/MS），可以测定 8-羟基-2'-脱氧鸟苷（8-OHdG）、邻-邻'-二酪氨酸（diY）、丙二醛（MDA）和四种 F-异前列腺素同型物：8-异前列腺素 F（8-PGF）、11β-前列腺素 F（11-PGF）、15（R）-前列腺素 F（15-PGF）和 8-异，15（R）-前列腺素 F（8,15-PGF）。DNPH 衍生化和 SPE 进行了优化，以提高分析目标化学物质的灵敏度和选择性。目标分析物在尿液中的检测限低于 30pg/mL。分析物的日内和日间变异系数低于相对标准偏差的 16%，目标化学物质添加到合成尿液中的回收率接近 100%。该方法应用于 21 个实际尿液样本的分析，8-PGF 和 15-PGF 的检出频率为 85%，11-PGF 为 71%，8,15-PGF 为 81%，diY、8-OHdG 和 MDA 为 100%。该方法选择性好、准确度高、精密度高，可同时测定尿液样品中多种不同分子来源的 OSB。
