International Institute for Translational Chinese Medicine, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, 510006, PR China.
International Institute for Translational Chinese Medicine, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, 510006, PR China.
Anal Biochem. 2021 May 15;621:114153. doi: 10.1016/j.ab.2021.114153. Epub 2021 Mar 6.
Sample preparation plays a crucial part in plasma metabolomics. In order to obtain an optimal sample extraction method for gas chromatography mass spectrometry (GC-MS)-based plasma metabolomics, five different extraction strategies including protein precipitation, liquid-liquid extraction and solid-phase extraction were evaluated systematically for both plasma untargeted- and targeted-metabolomics. The comprehensive evaluation revealed that the all-in-one sample preparation method, MeOH-MTBE-HO (1:5:1.5, v/v/v), was the optimal extraction method for both untargeted- and targeted-metabolomics. Next, the optimal sample preparation protocol was applied in plasma metabolomics of osteoarthritis (OA). A panel containing cholesterol, lactic acid, stearic acid, alpha-tocopherol and oxalic acid was selected as candidate biomarker to distinguish OA patients from healthy controls (HC) based on the support vector machine (SVM) classification model. The discriminating capability of the candidate biomarker panel was further validated successfully with logistic regression and principal components analysis (PCA) analysis. Therefore, the panel could potentially act as diagnostic biomarker for osteoarthritis.
样品制备在血浆代谢组学中起着至关重要的作用。为了获得基于气相色谱-质谱联用（GC-MS）的血浆代谢组学的最佳样品提取方法，我们系统地评估了包括蛋白质沉淀、液-液萃取和固相萃取在内的五种不同的提取策略，用于非靶向和靶向代谢组学。综合评价表明，MeOH-MTBE-HO（1:5:1.5，v/v/v）的一体化样品制备方法是用于非靶向和靶向代谢组学的最佳提取方法。接下来，我们将最优的样品制备方案应用于骨关节炎（OA）的血浆代谢组学研究。基于支持向量机（SVM）分类模型，我们选择了一个包含胆固醇、乳酸、硬脂酸、α-生育酚和草酸的候选生物标志物panel，用于区分 OA 患者和健康对照（HC）。候选生物标志物 panel 的判别能力还通过逻辑回归和主成分分析（PCA）分析得到了成功验证。因此，该 panel 可能是骨关节炎的潜在诊断生物标志物。
