Amino Acid Metabolomics Using LC MS/MS: Assessment of Cancer Cell Resistance in a Simulated Tumor Microenvironment.
We performed a comprehensive quantification of 20 amino acids in RPMI 1640 medium cultured human colorectal adenocarcinoma cells to evaluate the efficacy of 5 fluorouracil treatment under hypoxic and hypoglycemic conditions, which mimic the tumor microenvironment. In this study, we developed a simple and comprehensive analytical method by using LC MS/MS connected to the Intrada amino acid column, which eluted amino acids within 9 min. The present method covered a linearity range of 3.6 1818 mM, except for Gly (227 1818 mM) Ala, Asp, His (7.1 1818 mM each) and Trp (3.6 909 mM) The limits of detection were in the range of 0.02 38.0 pmol per injection in a standard solution. Amino acid concentration data were analyzed using principal component analysis to represent samples on two dimensional graphs. Linear discriminant analysis was used to classify samples on the score plots. Using this approach, the effect of 5 fluorouracil treatment could be successfully discriminated at high discrimination rates. Moreover, several amino acids were extracted from corresponding loading plots as candidate markers for distinguishing the effects of the 5 fluorouracil treatment or tumor microenvironmental conditions. These results suggest that our proposed method might be a useful tool for evaluating the efficacy of anticancer drugs in the tumor microenvironment.
Analytical sciences the international journal of the Japan Society for Analytical Chemistry