Zhang R, Wang J, Yuan G, Wei C, Liu X, Wang B, Gao H, Guo R
Institute of Clinical Pharmacology, Qilu Hospital of Shandong University, Jinan, Shandong, People's Republic of China.
Arzneimittelforschung. 2012 Jun;62(6):290-4. doi: 10.1055/s-0032-1308980. Epub 2012 Apr 2.
The purpose of this study is to determine the concentrations of norcantharidin (CAS NO: 5442-12-6) in mouse tissues and investigate its tissue distribution after intragastric administration of disodium norcantharidate solution. A highly sensitive and specific liquid chromatography-tandem mass spectrometric (LC-MS/MS) method was developed and validated, using ribavirin (CAS NO: 36791-04-5) as the internal standard (IS). Norcantharidin and IS were extracted from 0.3 mL tissue homogenates using protein precipitation with acetone under acid condition. The analyte was separated on a C18 reverse phase column and analyzed by MS/MS in the multiple reaction monitoring (MRM) mode using ESI with positive ionization, m/z 169→123 for norcantharidin and m/z 267→135 for IS. The developed method was validated over a linear range of concentrations 0.015 μg·mL - 1 in liver, lung, kidney, stomach, small intestine, uterus and testis, 0.0050.5 μg·mL - 1 in heart, spleen and brain, the correlation coefficients (r2) were between 0.9918 and 0.9976. The tissue distribution study result was as follows: The AUC0-t of norcantharidin in tissues was in the order as follows: small intestine, stomach, uterus, kidney, testis, liver, lung, spleen, heart, brain.
本研究的目的是测定小鼠组织中去甲斑蝥素（CAS编号：5442-12-6）的浓度，并研究灌胃给予去甲斑蝥酸钠溶液后其在组织中的分布情况。建立并验证了一种高灵敏度和特异性的液相色谱-串联质谱（LC-MS/MS）方法，使用利巴韦林（CAS编号：36791-04-5）作为内标（IS）。在酸性条件下，通过丙酮蛋白沉淀法从0.3 mL组织匀浆中提取去甲斑蝥素和内标。分析物在C18反相柱上分离，并采用电喷雾电离正离子模式，在多反应监测（MRM）模式下通过MS/MS进行分析，去甲斑蝥素的m/z为169→123，内标的m/z为267→135。所建立的方法在肝脏、肺、肾脏、胃、小肠、子宫和睾丸中的线性浓度范围为0.015 μg·mL - 1，在心脏、脾脏和大脑中的线性浓度范围为0.0050.5 μg·mL - 1，相关系数（r2）在0.9918至0.9976之间。组织分布研究结果如下：去甲斑蝥素在各组织中的AUC0-t顺序如下：小肠、胃、子宫、肾脏、睾丸、肝脏、肺、脾脏、心脏、大脑。
