Evaluation of the Absorption of Methotrexate on Cells and Its Cytotoxicity Assay by Using an Integrated Microfluidic Device Coupled to a Mass Spectrometer

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• 作者：Dan Gao ; Haifang Li ; Niejun Wang ; Jin-Ming Lin
• 刊名：Analytical Chemistry
• 出版年：2012
• 出版时间：November 6, 2012
• 年：2012
• 卷：84
• 期：21
• 页码：9230-9237
• 全文大小：455K
• 年卷期：v.84,no.21(November 6, 2012)
• ISSN：1520-6882

文摘

An integrated microfluidic device was developed for high-throughput drug screening with an online electrospray ionization quadrupole time-of-flight mass spectrometer (ESI-Q-TOF MS). The multiple gradient generator followed by an array of microscale cell culture chambers and on-chip solid-phase extraction (SPE) columns for sample pretreatment prior to mass analysis was integrated in the device which was fabricated in one single step. By using the combination system, the process for characterization of drug absorption and evaluation of cytotoxicity could be simultaneously realized. To validate the feasibility, the absorption of methotrexate and its effects on HepG2 and Caco-2 cells were investigated. With the increasing concentration of drugs, the percentage of apoptotic cells appeared in a dose-dependent fashion. By comparison with the results obtained from ESI-Q-TOF MS analysis and cytotoxicity assay, we found that higher intracellular drug concentration resulted in increased cell cytotoxicity. The technique presented herein provides an easy protocol to screen drugs rapidly with low drug consumption, high throughput, and high sensitivity.