Functional Evaluation of Anticancer Drugs in Human Leukemia Cells Based on Metabolic Profiling Technique

摘要

To date, mass spectrometry (MS) coupled with pre-separation technique such as liquid chromatography (LC-MS) or gas chromatography (GC-MS) has been known to be a conventional strategy for metabolomics, but these methods have several disadvantages for pharmaceutical metabolomics. First, sample preparation of such methods includes complicated steps such as sample extraction, derivatization, desalting, and/or concentration before further MS analysis. Second, large sample volume is required because of lower sensitivity. Third, albeit with new LC developments such as ultra-performance liquid chromatography (UPLC), total LC analysis time is still on the order of several minutes. Although GC coupled with time-of-flight MS had been developed to take shortertime than conventional GC-quadrupole-MS because of the ability to deconvolute co-eluted peaks easily, analysis time of GC is almost the same as that of UPLC. There is another problem that thwarts the development of MS-based metabolomics techniques. The appearance of adduct ions containing sodium or potassium ion, especially in positive ion mode, makes metabolomic data more complicated. Resolving of these issues is indispensable for highly sensitive and high-throughput analysis in pharmaceutical metabolomics research area because thousands of samples should be analyzed in the first large scale screening. Thus, 9-aminoacridine (9-AA) was utilized in the present study to develop MS mearsurement with negative ionization mode. In contrast to the methods mentioned above, direct MS analysis, especially, matrix-assisted laser des-orption ionization (MALDI)-MS has advantages for metabolite analysis because it is a highly sensitive, high-throughput, and low sample-consuming (within 1 mu L) technique compared tothe other conventional analytical platforms based on LC-, GC-or CE-MS. In addition, MALDI has recently been reported to be suitable not only for high-molecular-weight polymers or peptides but for low-molecular-weight