Department of Pharmaceutical Analysis, School of Pharmacy, Second Military Medical University, Shanghai, P. R. China; College of Pharmacy, Fujian University of Traditional Chinese Medicine, Fuzhou, Fujian, P. R. China; Shanghai Key Laboratory for Pharmaceutical Metabolite Research, Shanghai, P. R. China.
J Sep Sci. 2013 Oct;36(19):3184-90. doi: 10.1002/jssc.201300451. Epub 2013 Sep 3.
A simple, rapid, high-throughput, and highly sensitive LC-MS/MS was developed to determine anisodamine in a small volume (50 μL) of beagle dog plasma using atropine sulfate as the internal standard. The analyte and internal standard were isolated from 50 μL plasma samples after a one-step protein precipitation using Sirocco 96-well protein precipitation filtration plates. The separation was accomplished on a Hanbon Hedera CN column (100 × 4.6 mm, 5 μm) and the run time was 4 min. A Micromass Quatro Ultima mass spectrometer equipped with an ESI source was operated in the multiple reaction monitoring mode with the precursor-to-product ion transitions m/z 306.0→140.0 (anisodamine) and 290.0→123.9 (atropine) used for quantitation. The method was sensitive with a low LOQ of 0.05 ng/mL, and good linearity in the range 0.05-50 ng/mL for anisodamine (r(2) ≥ 0.995). All the validation data, such as accuracy, intra- and interrun precision, were within the required limits. The method was successfully applied to the pharmacokinetic study of anisodamine hydrochloride injection in beagle dogs.
建立了一种灵敏、快速、高通量的 LC-MS/MS 方法，用于测定犬血浆中 50 μL 小体积的山莨菪碱，以内硫酸阿托品为内标。采用 Sirocco 96 孔蛋白沉淀过滤板一步法蛋白沉淀，从 50 μL 血浆样品中分离出分析物和内标。分离在 Hanbon Hedera CN 柱（100×4.6mm，5μm）上进行，运行时间为 4 分钟。采用 Micromass Quatro Ultima 质谱仪，配有 ESI 源，在多重反应监测模式下操作，前体到产物离子跃迁 m/z 306.0→140.0（山莨菪碱）和 290.0→123.9（阿托品）用于定量。该方法灵敏，LOQ 低至 0.05ng/mL，山莨菪碱在 0.05-50ng/mL 范围内线性良好（r（2）≥0.995）。所有验证数据，如准确度、日内和日间精密度均在要求范围内。该方法成功应用于犬盐酸山莨菪碱注射液的药代动力学研究。
