Department of Chemistry, Middle East Technical University, Ankara, Turkey.
Department of Metallurgical and Materials Engineering, Middle East Technical University, Ankara, Turkey.
J Sep Sci. 2020 May;43(9-10):1925-1933. doi: 10.1002/jssc.202000211. Epub 2020 Apr 3.
Recently the connection between oxidative stress and various diseases, including cancer and Alzheimer's, attracts notice as a pathway suitable for diagnostic purposes. 8-Oxo-deoxyguanosine and 8-oxo-deoxyadenosine produced from the interaction of reactive oxygen species with DNA become prominent as biomarkers. Several methods have been developed for their determination in biofluids, including solid-phase extraction and enzyme-linked immunosorbent assays. However, still, there is a need for reliable and fast analytical methods. In this context, solid-phase microextraction offers many advantages such as flexibility in geometry and applicable sample volume, as well as high adaptability to high-throughput sampling. In this study, a solid-phase microextraction method was developed for the determination of 8-oxo-deoxyguanosine and 8-oxo-deoxyadenosine in biofluids. The extractive phase of solid-phase microextraction consisted of hydrophilic-lipophilic balanced polymeric particles. In order to develop a solid-phase microextraction method suitable for the determination of the analytes in saliva and urine, several parameters, including desorption solvent, desorption time, sample pH, and ionic strength, were scrutinized. Analytical figures of merit indicated that the developed method provides reasonable interday and intraday precisions (<15% in both biofluids) with acceptable accuracy. The method provides a limit of quantification for both biomarkers at 5.0 and 10.0 ng/mL levels in saliva and urine matrices, respectively.
最近,氧化应激与各种疾病(包括癌症和阿尔茨海默病)之间的联系作为一种适合诊断目的的途径引起了人们的注意。活性氧与 DNA 相互作用产生的 8-氧代-脱氧鸟苷和 8-氧代-脱氧腺苷成为生物标志物的突出代表。已经开发出几种用于测定生物流体中这些物质的方法,包括固相萃取和酶联免疫吸附测定。然而,仍然需要可靠和快速的分析方法。在这种情况下,固相微萃取具有许多优点,例如几何形状和适用的样品体积灵活,以及对高通量采样的高度适应性。在这项研究中,开发了一种用于测定生物流体中 8-氧代-脱氧鸟苷和 8-氧代-脱氧腺苷的固相微萃取方法。固相微萃取的萃取相由亲水-亲脂平衡聚合物颗粒组成。为了开发一种适用于测定唾液和尿液中分析物的固相微萃取方法,考察了几种参数,包括洗脱溶剂、洗脱时间、样品 pH 值和离子强度。分析性能指标表明,该方法在唾液和尿液基质中对两种生物标志物提供了合理的日间和日内精密度(<15%)和可接受的准确度。该方法在唾液和尿液基质中分别为两种生物标志物提供了 5.0 和 10.0 ng/mL 水平的定量限。