Innovation Center for Medical Redox Navigation, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
Anal Chem. 2010 Jan 15;82(2):498-504. doi: 10.1021/ac901083a.
In the present study, a high-throughput and nontargeted metabolomic technique using matrix-assisted laser desorption ionization-mass spectrometry (MALDI-MS) was developed for the rapid analysis of cellular metabolites. Either the detection limit or the linearity between concentrations of several standards and peak intensities was examined, indicating a detection limit lower than 10 fmol/well with a high linearity at low concentrations. To verify the validity of this method, metabolites from human acute lymphoblastic leukemia Jurkat cells were analyzed. Only 2,500 cells suspended in PBS were directly dropped onto a stainless MALDI sample plate, followed by mixing with matrix on the sample plate. Up to 150 metabolite peaks were detected from a single analysis within 90 s. For multivariate analysis of Jurkat cells against drug-treatment, three anticancer drugs were utilized. Principal component analysis of metabolites showed clear independent clusters for cells treated with these anticancer drugs. Furthermore, several metabolites involved in nucleotide synthesis were found to contribute to the separation of each cluster. These data suggest that the high-throughput MALDI-MS-based metabolomic technique proposed in the present study can be utilized for drug screening and validation of drug efficacy and safety.
在本研究中，开发了一种基于基质辅助激光解吸电离质谱（MALDI-MS）的高通量非靶向代谢组学技术，用于快速分析细胞代谢物。我们检查了几种标准品的检测限或浓度与峰强度之间的线性关系，结果表明检测限低于 10fmol/孔，在低浓度下具有很高的线性度。为了验证该方法的有效性，我们分析了人急性淋巴细胞白血病 Jurkat 细胞的代谢物。仅将悬浮在 PBS 中的 2500 个细胞直接滴加到不锈钢 MALDI 样品板上，然后在样品板上与基质混合。在 90 秒内，单次分析可检测到多达 150 个代谢物峰。对于 Jurkat 细胞对抗癌药物治疗的多变量分析，我们使用了三种抗癌药物。代谢物的主成分分析显示，用这些抗癌药物处理的细胞有明显的独立聚类。此外，还发现几种参与核苷酸合成的代谢物有助于每个聚类的分离。这些数据表明，本研究中提出的基于高通量 MALDI-MS 的代谢组学技术可用于药物筛选以及验证药物的疗效和安全性。
