Biochemistry and Toxicology Laboratory, Lyon Sud Hospital, University Hospital of Lyon, CEDEX, 69495 Pierre-Bénite, France.
Inserm U1052, CNRS UMR5286 Centre de Recherche en Cancérologie de Lyon, 69000 Lyon, France.
Int J Mol Sci. 2023 Feb 13;24(4):3770. doi: 10.3390/ijms24043770.
Performances of metabolomic methods have been widely studied on biological matrices such as serum, plasma, and urine; but much less on in vitro cell extracts. While the impact of cell culture and sample preparation on results are well-described, the specific effect of the in vitro cellular matrix on the analytical performance remains uncertain. The aim of the present work was to study the impact of this matrix on the analytical performance of an LC-HRMS metabolomic method. For this purpose, experiments were performed on total extracts from two cell lines (MDA-MB-231 and HepaRG) using different cell numbers. Matrix effects, carryover, linearity, and variability of the method were studied. Results showed that the performances of the method depend on the nature of the endogenous metabolite, the cell number, and the nature of the cell line. These three parameters should, therefore, be considered for the processing of experiments and the interpretation of results depending on whether the study focuses on a limited number of metabolites or aims to establish a metabolic signature.
代谢组学方法的性能已在血清、血浆和尿液等生物基质上得到了广泛研究；但在体外细胞提取物上的研究却少得多。虽然细胞培养和样品制备对结果的影响已有详细描述，但细胞外基质对分析性能的具体影响仍不确定。本工作的目的是研究该基质对 LC-HRMS 代谢组学方法分析性能的影响。为此，使用不同的细胞数量，在两种细胞系（MDA-MB-231 和 HepaRG）的总提取物上进行了实验。研究了基质效应、交叉污染、线性和方法的变异性。结果表明，方法的性能取决于内源性代谢物的性质、细胞数量和细胞系的性质。因此，应根据研究是侧重于有限数量的代谢物还是旨在建立代谢特征，考虑这三个参数来处理实验和解释结果。
