A 384-well solid-phase extraction for LC/MS/MS determination of methotrexate and its 7-hydroxy metabolite in human urine and plasma
A 384-well solid-phase extraction for LC/MS/MS determination of methotrexate and its 7-hydroxy metabolite in human urine and plasma
复制标题
DOI：
10.1021/ac000897i
复制
发表时间：
2001-02-01
影响因子：
7.4
通讯作者：
Henion, J
中科院分区：
文献类型：
--
作者：
Rule, G;Chapple, M;Henion, J
已开发了用于人尿液和血浆中甲氨蝶呤及其主要代谢产物7-羟基甲氨蝶呤的384孔板固相萃取程序。这种形式以前没有用于从生物流体中固相萃取药物。384孔板含有与二氧化硅颗粒结合的C-18固定相，二氧化硅颗粒被掺入玻璃纤维膜中。在尿液中，甲氨蝶呤和7-羟基甲氨蝶呤的定量曲线范围分别为1 - 50 μ g/mL和50 - 1000 ng/mL，在血浆中分别为5 - 250 ng/mL和5 - 100 ng/mL。使用20-穆尔样品等分试样通过线性回归定量两种分析物。进行了评价颗粒大小、洗脱体积和进样体积对信号强度影响的实验，并将其与检查威尔斯之间交叉污染的实验结果沿着报告。测定尿液中的回收率大于或等于95%。在14小时内分析的384个样品的运行结果表明，384孔SPE可以成功地利用，以增加分析运行的大小和样品通量的LC/MS/MS测定生物样品中的小药物分子。
A solid-phase extraction procedure, in a 384-well format, has been developed for methotrexate and its primary metabolite, 7-hydroxymethotrexate, in human urine and plasma. This format has not been utilized previously for solid-phase extraction of drugs from biological fluids, The 384-well plates contained a C-18 stationary phase bonded to silica particles which are incorporated into a glass-fiber membrane. Methotrexate and 7-hydroxymethotrexate have been quantified across the curve range of 1 to 50 mug/mL and 50 to 1000 ng/mL, respectively, in urine and from 5 to 250 ng/mL and 5 to 100 ng/mL, respectively, in plasma. Both analytes are quantified by linear regression using 20-muL sample aliquots. Experiments to evaluate the influence of particle size, elution volume, and injection volume on signal intensity were conducted and are reported, along with the results of experiments examining cross contamination between wells. Recovery was determined to be greater than or equal to 95% from urine. Results from a run of 384 samples analyzed over a 14-h period indicate that 384-well SPE can be successfully utilized to increase analytical run sizes and sample throughput for LC/MS/MS determination of small drug molecules in biological samples.