The State Key Laboratory of Natural Medicines and Department of Pharmacognosy, China Pharmaceutical University, 210038, Nanjing, China; The Ministry of Education (MOE) Key Laboratory for Standardization of Chinese Medicines and the State Administration of TCM (SATCM) Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, 201210, Shanghai, China; School of Pharmacy, Jiangxi Science and Technology Normal University, 330013, Nanchang, China.
J Appl Toxicol. 2014 Feb;34(2):149-57. doi: 10.1002/jat.2844. Epub 2012 Dec 12.
In this study, we have developed and validated a simple, accurate and sensitive gas chromatography-mass spectrometry (GC-MS) method for simultaneous quantification of 18 fatty acids in rat serum, including both non-esterified (NEFA) and esterified (EFA) fatty acids, and subsequent analysis of fatty acid metabolic profiles. This novel method was used to evaluate the serum levels of fatty acids from vehicle- and acetaminophen (APAP)-treated rats. Serum levels of 7 NEFAs and 14 EFAs were significantly higher in APAP-treated rats 24 h after APAP administration at 1500 mg kg⁻¹ when compared with vehicle-treated controls. Control and APAP-treated rats could be differentiated based on their metabolic profiles using two different chemometric analysis methods: principle component analysis (PCA) and partial least squares-discriminant analysis (PLS-DA). More importantly, we identified the following NEFAs as potential biomarkers of APAP-induced liver injury: oleic acid (C18:1n9), linoleic acid (C18:2n6), docosahexaenoic acid (C22:6n3) and arachidonic acid (C20:4n6). The serum concentrations of C18:1n9, C18:2n6 and C22:6n3 were all positively correlated (r > 0.8; Pearson's correlation analysis) with the activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST). These results suggest that a novel targeted metabolomics method based on the metabolic profiling of fatty acids analyzed by GC-MS provides exact serum concentrations of fatty acids as well as a prospective methodology to evaluate chemically induced hepatotoxicity.
在这项研究中，我们开发并验证了一种简单、准确、灵敏的气相色谱-质谱（GC-MS）方法，用于同时定量分析大鼠血清中的 18 种脂肪酸，包括非酯化（NEFA）和酯化（EFA）脂肪酸，并随后分析脂肪酸代谢谱。该新方法用于评估乙酰氨基酚（APAP）处理和对照大鼠的血清脂肪酸水平。与对照相比，在给予 1500mg/kg APAP 后 24 小时，APAP 处理的大鼠血清中 7 种 NEFA 和 14 种 EFA 的水平明显更高。使用两种不同的化学计量学分析方法：主成分分析（PCA）和偏最小二乘判别分析（PLS-DA），可以根据代谢谱区分对照和 APAP 处理的大鼠。更重要的是，我们确定了以下 NEFA 作为 APAP 诱导肝损伤的潜在生物标志物：油酸（C18:1n9）、亚油酸（C18:2n6）、二十二碳六烯酸（C22:6n3）和花生四烯酸（C20:4n6）。C18:1n9、C18:2n6 和 C22:6n3 的血清浓度均与丙氨酸氨基转移酶（ALT）和天冬氨酸氨基转移酶（AST）的活性呈正相关（r>0.8；Pearson 相关分析）。这些结果表明，基于 GC-MS 分析的脂肪酸代谢谱的新型靶向代谢组学方法可提供准确的脂肪酸血清浓度，并为评估化学诱导肝毒性提供了有前景的方法。
