Departamento de Química, ICEx, Universidade Federal de Minas Gerais, Avenida Antônio Carlos, 6627-31270901, Belo Horizonte, MG, Brazil.
Nuclear Technology Development Center/National Nuclear Energy Commission (CDTN/CNEN), Belo Horizonte, MG, Brazil.
Anal Biochem. 2020 May 15;597:113666. doi: 10.1016/j.ab.2020.113666. Epub 2020 Mar 3.
Metabolomic studies are essential to identify and quantify key metabolites in biological systems. Analysis of amino acids (AA) is very important in target metabolomics studies. Chromatographic methods are used to support metabolite determinations. Therefore, this work presents analysis of 17 AA in Saccharomyces cerevisiae cells (a useful model in the study of cancer metabolism) exposed to sodium selenite and gamma radiation. An improved GC/MS method using propyl chloroformate/propanol as derivatizing reagent was applied to AA determinations. The method exhibited good linearity in the range of 0.08-600.00 mg L; limits of determination from 0.04 to 1.60 mg L; limits of quantification from 0.08 to 2.76 mg L; repeatability ranging from 1.9 to 11.4 %; and precision ranging from 2.8 to 13.8 %. The correlations between selenite/gamma radiation with AA profile was investigated to establish candidates for cancer biomarkers. The analyses of yeast cultures found high concentrations of amino acids, such as Alanine, Serine, Glutamate, and Lysine, which might be associated with the development of metabolic adaptations of cancer based on its high demand for biomass and energy, found both in this model and neoplastic cells.
代谢组学研究对于鉴定和量化生物系统中的关键代谢物至关重要。在目标代谢组学研究中,分析氨基酸(AA)非常重要。色谱方法用于支持代谢物的测定。因此,本工作分析了暴露于亚硒酸钠和γ辐射的酿酒酵母细胞(癌症代谢研究的有用模型)中的 17 种 AA。应用丙基氯甲酸酯/丙醇作为衍生试剂的改进 GC/MS 方法用于 AA 的测定。该方法在 0.08-600.00 mg/L 的范围内表现出良好的线性关系;测定下限为 0.04-1.60 mg/L;定量下限为 0.08-2.76 mg/L;重复性范围为 1.9-11.4%;精密度范围为 2.8-13.8%。研究了亚硒酸钠/γ辐射与 AA 谱之间的相关性,以确定癌症生物标志物的候选物。对酵母培养物的分析发现了高浓度的氨基酸,如丙氨酸、丝氨酸、谷氨酸和赖氨酸,这可能与癌症基于其对生物量和能量的高需求的代谢适应的发展有关,在这个模型和肿瘤细胞中都有发现。