[Classification of anticancer drugs with different mechanisms based on amino-acid consumption profiling in culture media]

An approach for quantitative determination of amino-acid consumption profiling in culture media by high performance liquid chromatography with fluorescence detection (HPLC-FLD) was developed and validated, using o-phthalic dicarboxaldehyde (OPA) as the derivatizing reagent and norvaline as the internal standard. Mobile phase A was 10 mmol/L Na2HPO4Na2B4O7 buffer (pH 7.95), and mobile phase B was acetonitrile-methanol-water (45:45:10, v/v/v). The linear elution program was 5% B at the start and 52% B at the end in 35 min. The 17 free amino-acids (FAAs) were separated satisfactorily in 33 min. Following HeLa cells incubation in conditioned medias of taxol (4 micromol/L) and mitomycin (75 micromol/L), respectively, with control for 24 h, the media 17 amino-acid consumption profilings were determined, and then analyzed by multivariate statistical analysis based on Matlab7.1 software platform. Relation analysis performed by partial least squares-discriminant analysis (PLS-DA) indicated that in comparison with the control group, the media amino-acid consumption profiling can distinguish the two anticancer drugs with different mechanisms, which provides a new perspective for the pre-classification of drug action mechanisms during the screening of new anticancer drugs. Meanwhile, the idea from the outer into the inner has convenient and economic characteristics.