建立并验证了一种使用固相萃取同时测定人血浆中咪达唑仑及其 1'-羟基代谢物浓度的液相色谱-串联质谱方法。
Development and validation of a method using supported liquid extraction for the simultaneous determination of midazolam and 1'-hydroxy-midazolam in human plasma by liquid chromatography with tandem mass spectrometry detection.
机构信息
Department of Chemistry and Pharmaceutics, Active Biotech AB, Box 724, SE-22007 Lund, Sweden.
出版信息
J Pharm Biomed Anal. 2012 Jan 25;58:71-7. doi: 10.1016/j.jpba.2011.09.015. Epub 2011 Sep 22.
The metabolic conversion of midazolam (MDZ) to its main metabolite 1'-hydroxy-midazolam (1-OH-MDZ) can be used as a probe drug for cytochrome P450 3A (CYP3A) activity. A sensitive method for the simultaneous determination of MDZ and its metabolite 1-OH-MDZ in human plasma using supported liquid extraction (SLE) in combination with liquid chromatography-tandem mass spectrometry (LC-MS/MS) detection was developed and validated. Plasma samples (100 μL) were diluted with 0.5M NH(3) (aq) containing deuterated internal standards. The samples were extracted with ethyl acetate on a 96-well SLE-plate. Separation was performed on a Symmetry Shield RP18 column using an acidic gradient running from 2% to 95% methanol in 3 min. Detection was performed using a triple quadrupole mass spectrometer running in positive electrospray selected reaction monitoring (SRM) mode. The validated dynamic range was 0.2-100 nmol/L for both analytes. In the concentration range 0.6-75 nmol/L the extraction recoveries were in the ranges 91.2-98.6% and 94.5-98.3% for MDZ and 1-OH-MDZ, respectively. Matrix effects were more pronounced for MDZ than for 1-OH-MDZ but the response was still 75.4% or higher compared to a reference. The overall repeatability was within 2.2-7.6% for both analytes, the overall reproducibility was within 3.1-10.2% for both analytes and the overall accuracy bias was within -1.1 to 7.5% for both analytes. The method was successfully applied to determine the plasma concentrations of MDZ and 1-OH-MDZ in 14 healthy volunteers up to 24h after administration of a single oral dose of 2mg MDZ. The SLE technology was found to be convenient and suitable for sample preparation, and the developed method was found to be rapid, selective and reproducible for the simultaneous determination of MDZ and 1-OH-MDZ in human plasma.
咪达唑仑（MDZ）向其主要代谢产物 1'-羟基咪达唑仑（1-OH-MDZ）的代谢转化可用作细胞色素 P450 3A（CYP3A）活性的探针药物。建立并验证了一种灵敏的人血浆中 MDZ 及其代谢物 1-OH-MDZ 的同时测定方法，该方法采用固相萃取（SLE）结合液相色谱-串联质谱（LC-MS/MS）检测。将 100 μL 血浆样品用含氘代内标的 0.5M NH 3 (aq) 稀释。样品在 96 孔 SLE 板上用乙酸乙酯萃取。在 Symmetry Shield RP18 柱上，采用酸性梯度，在 3 分钟内从 2%到 95%甲醇洗脱，进行分离。采用正电喷雾选择反应监测（SRM）模式的三重四极杆质谱仪进行检测。两种分析物的验证有效范围均为 0.2-100 nmol/L。在 0.6-75 nmol/L 的浓度范围内，MDZ 和 1-OH-MDZ 的提取回收率分别为 91.2-98.6%和 94.5-98.3%。基质效应对于 MDZ 比 1-OH-MDZ 更为明显，但与参比相比，其响应仍为 75.4%或更高。两种分析物的总体重复性均在 2.2-7.6%范围内，两种分析物的总重现性均在 3.1-10.2%范围内，两种分析物的总准确度偏差均在-1.1 至 7.5%范围内。该方法成功地应用于 14 名健康志愿者单次口服 2mg MDZ 后 24 小时内测定血浆中 MDZ 和 1-OH-MDZ 的浓度。发现固相萃取技术方便且适用于样品制备，所建立的方法快速、选择性好、重现性好，可用于人血浆中 MDZ 和 1-OH-MDZ 的同时测定。
Zotero 插件