Department of Medicinal Chemistry, College of Pharmacy, University of Sharjah, Sharjah 27272, United Arab Emirates.
Research Institute of Medical and Health Sciences, University of Sharjah, Sharjah 27272, United Arab Emirates.
Int J Mol Sci. 2023 Jan 10;24(2):1354. doi: 10.3390/ijms24021354.
Untargeted multi-omics analysis of plasma is an emerging tool for the identification of novel biomarkers for evaluating disease prognosis, and for developing a better understanding of molecular mechanisms underlying human disease. The successful application of metabolomic and proteomic approaches relies on reproducibly quantifying a wide range of metabolites and proteins. Herein, we report the results of untargeted metabolomic and proteomic analyses from blood plasma samples following analyte extraction by two frequently-used solvent systems: chloroform/methanol and methanol-only. Whole blood samples were collected from participants (n = 6) at University Hospital Sharjah (UHS) hospital, then plasma was separated and extracted by two methods: (i) methanol precipitation and (ii) 4:3 methanol:chloroform extraction. The coverage and reproducibility of the two methods were assessed by ultra-high-performance liquid chromatography-electrospray ionization quadrupole time-of-flight mass spectrometry (UHPLC-ESI-QTOF-MS). The study revealed that metabolite extraction by methanol-only showed greater reproducibility for both metabolomic and proteomic quantifications than did methanol/chloroform, while yielding similar peptide coverage. However, coverage of extracted metabolites was higher with the methanol/chloroform precipitation.
非靶向多组学分析血浆是一种新兴的工具，可用于鉴定评估疾病预后的新型生物标志物，并深入了解人类疾病的分子机制。代谢组学和蛋白质组学方法的成功应用依赖于可重现地定量检测广泛的代谢物和蛋白质。本文报告了两种常用溶剂系统（氯仿/甲醇和甲醇）提取分析物后，来自沙迦大学医院（UHS）参与者血液血浆样本的非靶向代谢组学和蛋白质组学分析结果。采集全血样本，然后通过两种方法分离和提取血浆：（i）甲醇沉淀和（ii）4:3 甲醇/氯仿提取。通过超高效液相色谱-电喷雾电离四极杆飞行时间质谱（UHPLC-ESI-QTOF-MS）评估了这两种方法的覆盖度和重现性。研究表明，与甲醇/氯仿相比，仅用甲醇提取代谢物在代谢组学和蛋白质组学定量方面具有更好的重现性，而产生的肽覆盖率相似。然而，甲醇/氯仿沉淀提取的代谢物覆盖度更高。
