Lee Xiao-Pen, Shouji Yukiko, Kumazawa Takeshi, Hasegawa Chika, Fujishiro Masaya, Sato Junichi, Hasegawa Iwao, Sato Keizo
Department of Legal Medicine, Showa University School of Medicine, Tokyo 142-8555, Japan.
Department of Legal Medicine, Showa University School of Medicine, Tokyo 142-8555, Japan.
Leg Med (Tokyo). 2017 Jan;24:36-55. doi: 10.1016/j.legalmed.2016.11.005. Epub 2016 Nov 27.
A high-throughput method was developed for the detection of 31 benzodiazepine drugs and tandospirone in human plasma by on-line column-switching ultra-fast liquid chromatography-tandem mass spectrometry. Plasma samples (100μl) spiked with the 32 drugs and oxazepam-d (internal standard) were diluted with 300μl of 13.3mM ammonium acetate/acetonitrile (33:67, v/v). After centrifugation and filtration, the clear supernatant was injected directly onto the extraction column (Oasis HLB cartridge column). The following procedure was fully automated. The analytes retained on the extraction column were eluted by backflushing of the extraction column and introduced into an analytical column (SUMIPAX ODS D-Swifter column, 30mm×3.0mm i.d.; particle size 2μm) by column switching. Quantification was performed by multiple reaction monitoring with positive-ion electrospray ionization. Distinct peaks appeared for each drug and the internal standard on each channel within 7min, including the extraction time. All drugs spiked into plasma showed recoveries of 83-95%. The regression equations for the 32 drugs showed excellent linearities in the range of 50-2000pg/ml of plasma and the limits of detection ranged from 20 to 50pg/ml. The lower and upper limits of quantitation were 50-100ng/ml and 2000pg/ml, respectively. Intra- and interday coefficients of variation for none of the drugs were greater than 13.6%. The accuracies of quantitation were 87-112%. The multiple reaction monitoring information-dependent acquisition of enhanced product ions method enabled the quantification and confirmation of diazepam, triazolam, and lorazepam obtained from actual plasma.
建立了一种高通量方法，用于通过在线柱切换超快速液相色谱-串联质谱法检测人血浆中的31种苯二氮䓬类药物和坦度螺酮。将加有32种药物和奥沙西泮-d（内标）的血浆样品（100μl）用300μl 13.3mM醋酸铵/乙腈（33:67，v/v）稀释。离心和过滤后，将清澈的上清液直接注入萃取柱（Oasis HLB柱）。以下步骤完全自动化。保留在萃取柱上的分析物通过萃取柱反冲洗脱，并通过柱切换引入分析柱（SUMIPAX ODS D-Swifter柱，30mm×3.0mm内径；粒径2μm）。通过正离子电喷雾电离的多反应监测进行定量。包括萃取时间在内，每种药物和内标在每个通道上7分钟内都出现了明显的峰。所有添加到血浆中的药物回收率为83-95%。32种药物的回归方程在血浆浓度50-2000pg/ml范围内显示出良好的线性，检测限为20-50pg/ml。定量下限和上限分别为50-100ng/ml和2000pg/ml。所有药物的日内和日间变异系数均不大于13.6%。定量准确度为87-112%。多反应监测信息依赖增强产物离子采集方法能够对实际血浆中获得的地西泮、三唑仑和劳拉西泮进行定量和确证。
