Unit of Integrative Metabolomics, Institute of Environmental Medicine, Karolinska Institutet, Stockholm, Sweden; Metabolomics Laboratory, Clinical Research Center, Medical University of Bialystok, 15-276 Bialystok, Poland.
Unit of Integrative Metabolomics, Institute of Environmental Medicine, Karolinska Institutet, Stockholm, Sweden.
Prostaglandins Other Lipid Mediat. 2024 Feb;170:106789. doi: 10.1016/j.prostaglandins.2023.106789. Epub 2023 Oct 23.
Urinary eicosanoid concentrations reflect inflammatory processes in multiple diseases and have been used as biomarkers of disease as well as suggested for patient stratification in precision medicine. However, implementation of urinary eicosanoid profiling in large-scale analyses is restricted due to sample preparation limits. Here we demonstrate a single solid-phase extraction of 300 µL urine in 96-well-format for prostaglandins, thromboxanes, isoprostanes, cysteinyl-leukotriene E and the linoleic acid-derived dihydroxy-octadecenoic acids (9,10- and 12,13-DiHOME). A simultaneous screening protocol was also developed for cortisol/cortisone and 7 exogenous steroids as well as 3 cyclooxygenase inhibitors. Satisfactory performance for quantification of eicosanoids with an appropriate internal standard was demonstrated for intra-plate analyses (CV = 8.5-15.1%) as well as for inter-plate (n = 35) from multiple studies (CV = 22.1-34.9%). Storage stability was evaluated at - 20 °C, and polar tetranors evidenced a 50% decrease after 5 months, while the remaining eicosanoids evidenced no significant degradation. All eicosanoids were stable over 3.5-years in urine stored at - 80 °C. This method will facilitate the implementation of urinary eicosanoid quantification in large-scale screening.
尿中二十烷类浓度反映了多种疾病中的炎症过程，已被用作疾病的生物标志物，并被建议用于精准医学中的患者分层。然而，由于样品制备的限制，尿二十烷类谱分析在大规模分析中的实施受到限制。在这里，我们展示了一种在 96 孔格式中单次固相萃取 300μL 尿液以同时检测前列腺素、血栓烷、异前列烷、半胱氨酰白三烯 E 和亚油酸衍生的二羟十八烯酸（9,10- 和 12,13-DiHOME）的方法。还同时开发了一种用于皮质醇/皮质酮和 7 种外源性类固醇以及 3 种环氧化酶抑制剂的同时筛选方案。结果表明，对于使用适当内标进行定量的二十烷类，无论是在板内分析（CV = 8.5-15.1%）还是在来自多个研究的板间分析（n = 35，CV = 22.1-34.9%），都具有良好的性能。在 -20°C 下评估了储存稳定性，极性四氢壬烷在 5 个月后下降了 50%，而其余的二十烷类则没有明显的降解。在 -80°C 储存的尿液中，所有的二十烷类都能稳定 3.5 年以上。该方法将促进尿二十烷类定量在大规模筛选中的应用。
