Measurement of intracellular accumulation of anthracyclines in cancerous cells by direct injection of cell lysate in MEKC/LIF detection
Measurement of intracellular accumulation of anthracyclines in cancerous cells by direct injection of cell lysate in MEKC/LIF detection
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DOI：
10.1002/elps.200900659
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发表时间：
2010-04
期刊：
影响因子：
2.9
通讯作者：
Julius Mbuna;T. Kaneta;T. Imasaka
中科院分区：
文献类型：
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作者：
Julius Mbuna;T. Kaneta;T. Imasaka
蒽环类是广泛用于治疗各种类型的实体癌和白血病的化疗药物。在此，我们报告了一种新的分析方法，蒽环类药物的细胞内积累使用MEKC/LIF检测。一个含水的分离系统允许细胞裂解液直接注入毛细管。MEKC迁移溶液由含有牛磺脱氧胆酸钠、（2-羟丙基）-γ-CD和SDS的pH 9.22硼酸盐缓冲液组成。用Nd：YAG激光（532 nm）或氩离子激光（488 nm）激发，用激光诱导荧光法检测蒽环类药物、阿霉素（DOX）和表阿霉素（EPI）。两种细胞系，人肱骨肿瘤细胞（RDES）和人肺肿瘤细胞（A549），用两种蒽环类抗生素的混合物处理固定的时间，然后通过直接注射细胞裂解物测定细胞内浓度。DOX和EPI的回收率为96.0-100.8%。在50至500 nM的浓度范围内，通过RSD定量的重现性小于3.9%日内和6.7%日间。对于A549，发现EPI的摄取略低于DOX的摄取，但在RDES中观察到更高水平的EPI。蒽环类抗生素在RDES中的细胞内蓄积大于A549，但两种类型的细胞在12 h后均分泌蒽环类抗生素。这些结果表明，MEKC与水介质是有用的研究细胞内摄取和积累的药物，因为细胞裂解物可以直接使用，没有预处理，如脱蛋白或溶剂提取的分析物。
Anthracyclines are chemotherapeutic drugs that are broadly used in the treatment of various types of solid cancers and leukemia. Herein, we report on a novel analytical method for intracellular accumulation of anthracyclines using MEKC/LIF detection. An aqueous separation system permitted the injection of cell lysates directly into the capillary. The MEKC migrating solution was made up of borate buffer at pH 9.22 containing sodium taurodeoxycholate, (2‐hydroxypropyl)‐γ‐CD, and SDS. The anthracyclines, Doxorubicin (DOX) and epirubicin (EPI) were detected by LIF using a Nd:YAG laser (532 nm) or an argon ion laser (488 nm) for excitation. Two cell lines, human humerus tumor cells (RDES) and human lung tumor cells (A549), were treated with a mixture of the two anthracyclines for fixed periods of time, and then intracellular concentrations were determined by injecting cell lysates directly. Recovery values of 96.0–100.8% were obtained for DOX and EPI. Reproducibility quantified by RSD was less than 3.9% intraday and 6.7% interday at concentrations ranging between 50 and 500 nM. The uptake of EPI was found to be slightly less than that of DOX for A549, but higher levels of EPI were observed in RDES. Intracellular accumulation of anthracyclines was greater in RDES than in A549, but both types of cells excreted anthracyclines after 12 h. These results indicate that MEKC with an aqueous medium is useful for investigating intracellular uptake and accumulation of drugs, since cell lysates can be used directly with no pretreatment such as deproteination or solvent extraction of analytes.