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.
[1]East China Normal Univ, Sch Chem & Mol Engn, Shanghai 200241, Peoples R China.
[2]Shanghai Jiao Tong Univ, Sch Pharm, Shanghai 200240, Peoples R China.
[3]Shanghai Univ Tradit Chinese Med, Ctr Chinese Med Therapy & Syst Biol, Shanghai 201203, Peoples R China.
[4]HD Biosci China Co Ltd, DMPK Dept, Shanghai 201201, Peoples R China.
[5]Xinjiang Med Univ, Affiliated Tumor Hosp, Dept Gynecol, Urumqi 830011, Peoples R China.
(8.0+极速模式)