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A simultaneously quantitative profiling method for 40 endogenous amino acids and derivatives in cell lines using hydrophilic interaction liquid chromatography coupled with tandem mass spectrometry

化学 色谱法 氨基酸 亲水作用色谱法 代谢组学 串联质谱法 液相色谱-质谱法 分析物 检出限 质谱法 电喷雾 高效液相色谱法 生物化学
作者
Bangjie Zhu,Li Li,Hai Wei,Wenbin Zhou,Junlian Yin,Fugang Li,Peiyuan Lin,Jiaqiang Sheng,Qingjiang Wang,Chao Yan,Yu Cheng
出处
期刊:Talanta [Elsevier]
卷期号:207: 120256-120256 被引量:24
标识
DOI:10.1016/j.talanta.2019.120256
摘要

Endogenous metabolites of amino acids and their derivatives in biosamples are frequently highlighted as the most differential metabolites in recent metabolomics studies. The method for the detection of amino acid derivatives such as N-acetyl amino acids and oligopeptides is rarely reported. We developed a rapid, high-throughput, sensitive and reliable quantitative method to simultaneously profile 40 underivatized amino acids and their derivatives including N-acetyl amino acids and oligopeptides in cell lines, based on ultra-high-performance liquid chromatography-electrospray tandem mass spectrometry (UHPLC- MS/MS) by using a hydrophilic interaction liquid chromatography (HILIC) column. The optimized method was successfully validated with satisfactory linearity, sensitivity, accuracy, precision, matrix effects, recovery and stability for all analytes. Only one compound (cysteine-glutathione disulfide) showed relatively low recoveries at three concentration levels (60.8–74.3%). The limit of quantification (LOQ) for most compounds was in the range of 0.6–10 ng/mL (6–100 pg on column). This method was successfully applied to the analysis of amino acids and their derivatives in breast cancer cell samples. Principal component analysis (PCA) and the orthogonal projections to latent structures (OPLS) showed a clear discrimination of the non-tumorigenic breast epithelial cell line MCF-10A from the breast cancer cell line HCC 1806. Characteristic metabolic changes in amino acid metabolism were observed in the breast cancer cell line. This quantified analytical method of 40 endogenous amino acids and their derivatives in cell lines meets the requirement of quantification in specific expanded metabolomics studies with good sensitivity.
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