Subcellular Quantification of Doxorubicin and Its Metabolite in Cultured Human Leukemia Cells Using Liquid Chromatography-Tandem Mass Spectrometry
Subcellular Quantification of Doxorubicin and Its Metabolite in Cultured Human Leukemia Cells Using Liquid Chromatography-Tandem Mass Spectrometry
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使用液相色谱-串联质谱法对培养的人白血病细胞中的阿霉素及其代谢物进行亚细胞定量
DOI：
10.1080/00032719.2012.680056
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发表时间：
2012
影响因子：
2
通讯作者：
Yun Chen
中科院分区：
文献类型：
--
作者：
Jinhui Xu;Yuan Liu;Y. Yu;Q. Ni;Yun Chen
多柔比星（DOX）作为一种抗癌药物在世界范围内广泛用于治疗白血病和实体瘤。然而，其临床应用在很大程度上受到心脏毒性的限制。DOX诱导的心脏毒性的最常提出的机制之一是代谢物的形成。然而，DOX细胞内代谢中涉及的酶途径迄今尚未完全阐明。为了更详细地描述DOX代谢，我们实验室开发了一种使用液相色谱-串联质谱法（LC/MS/MS）的测定方法，以同时测定亚细胞区室中的DOX及其主要代谢物多柔比星（DOXol）。使用固相萃取实现样品净化和富集，DOX和DOXol的经验证校准范围分别为5.00-1000 ng/mL和0.50-50.0 ng/mL。达到了良好的准确度和精密度。使用该测定，在两种人T白血病细胞系（即，Jurkat和CCRF-CEM）。还进行了时间进程分析。由此产生的DOX和DOXol亚细胞分布和浓度-时间曲线的变化可能归因于这些细胞系内还原酶的差异表达、活性和定位。更重要的是，这项工作表明，同时测定药物及其代谢产物在亚细胞区室可以实现使用LC/MS/MS。
Doxorubicin (DOX) is widely used in the world as an anticancer agent for the treatment of leukemia and solid tumors. However, its clinical use is largely limited by the emergence of cardiotoxicity. One of the most frequently proposed mechanisms for DOX induced cardiotoxicity is the formation of metabolites. However, the enzymatic pathways involved in DOX intracellular metabolism have not been fully elucidated thus far. To provide a more detailed description of DOX metabolism, an assay using liquid chromatography-tandem mass spectrometry (LC/MS/MS) was developed in our lab to simultaneously determine DOX and its primary metabolite doxorubicinol (DOXol) in subcellular compartments. Sample cleanup and enrichment were achieved using solid phase extraction, and the validated calibration ranges for DOX and DOXol were 5.00–1000 ng/mL and 0.50–50.0 ng/mL, respectively. Good accuracy and precision were achieved. Using this assay, the accumulation of DOX and DOXol in whole cells, nuclear-enriched fraction (NEF) and organelle-enriched fraction (OEF) were compared between two human T leukemia cell lines (i.e., Jurkat and CCRF-CEM). A time-course analysis was also carried out. The resulting varieties of DOX and DOXol subcellular distributions and concentration-time profiles might be attributed to the differential expression, activities, and localization of reductive enzymes within these cell lines. More importantly, this work demonstrated that simultaneous determination of drug and its metabolite in subcellular compartments could be achieved using LC/MS/MS.
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影响因子：
11.2
作者：
A. Bodley;Leroy F. Liu;M. Israel;R. Seshadri;Y. Koseki;F. Giuliani;S. Kirschenbaum;R. Silber
通讯作者：
A. Bodley;Leroy F. Liu;M. Israel;R. Seshadri;Y. Koseki;F. Giuliani;S. Kirschenbaum;R. Silber
DOI：
10.1073/pnas.85.10.3585
发表时间：
1988-05-01
影响因子：
11.1
作者：
OLSON, RD;MUSHLIN, PS;BOUCEK, RJ
通讯作者：
BOUCEK, RJ
影响因子：
7.4
作者：
Rabinowitz, Joshua D.;Kimball, Elizabeth
通讯作者：
Kimball, Elizabeth
DOI：
10.1016/j.jchromb.2004.04.030
发表时间：
2004-09-05
影响因子：
3
作者：
Arnold, RD;Slack, JE;Straubinger, RM
通讯作者：
Straubinger, RM