Sparreboom A, van Tellingen O, Nooijen W J, Beijnen J H
Department of Pharmacy, Slotervaart Hospital, Amsterdam, Netherlands.
J Chromatogr B Biomed Appl. 1995 Feb 17;664(2):383-91. doi: 10.1016/0378-4347(94)00495-q.
We have developed and validated a sensitive and selective assay for the quantification of paclitaxel and its metabolites 6 alpha, 3'-p-dihydroxypaclitaxel, 3'-p-hydroxypaclitaxel and 6 alpha-hydroxypaclitaxel in plasma, tissue, urine and faeces specimens of mice. Tissue and faeces were homogenized (approximately 0.1-0.2 g/ml) in bovine serum albumin (40 g/l) in water, and urine was diluted (1:5, v/v) in blank human plasma. Sample pretreatment involved liquid-liquid extraction of 200-1000 microliters of sample with diethyl ether followed by automated solid-phase extraction using cyano Bond Elut columns. 2'-Methylpaclitaxel was used as internal standard. The overall recovery of the sample pretreatment procedure ranged from 76 to 85%. In plasma, the lower limit of detection (LOD) and the lower limit of quantitation (LLQ) are 15 and 25 ng/ml, respectively, using 200 microliters of sample. In tissues, faeces and urine the LLQs are 25-100 ng/g, 125 ng/g and 25 ng/ml, respectively, using 1000 microliters (faeces: 200 microliters) of homogenized or diluted sample. The concentrations in the various biological matrices, for validation procedures spiked with known amounts of the test compounds, are read from calibration curves constructed in blank human plasma in the range 25-100,000 ng/ml for paclitaxel and 25-500 ng/ml for the metabolites. The accuracy and precision of the assay fall within the generally accepted criteria for bio-analytical assays.
我们已经开发并验证了一种灵敏且具选择性的分析方法，用于定量小鼠血浆、组织、尿液和粪便样本中的紫杉醇及其代谢物6α,3'-对羟基紫杉醇、3'-对羟基紫杉醇和6α-羟基紫杉醇。将组织和粪便在水中的牛血清白蛋白（40 g/l）中匀浆（约0.1 - 0.2 g/ml），尿液在空白人血浆中按1:5（v/v）稀释。样品预处理包括用乙醚对200 - 1000微升样品进行液 - 液萃取，随后使用氰基Bond Elut柱进行自动固相萃取。2'-甲基紫杉醇用作内标。样品预处理程序的总回收率在76%至85%之间。在血浆中，使用200微升样品时，检测下限（LOD）和定量下限（LLQ）分别为15和25 ng/ml。在组织、粪便和尿液中，使用1000微升（粪便：200微升）匀浆或稀释样品时，LLQ分别为25 - 100 ng/g、125 ng/g和25 ng/ml。对于用已知量的测试化合物加标的验证程序，各种生物基质中的浓度从空白人血浆中构建的校准曲线读取，紫杉醇的范围为25 - 100,000 ng/ml，代谢物的范围为25 - 500 ng/ml。该分析方法的准确度和精密度符合生物分析方法普遍接受的标准。
