College of Pharmacy, Gachon University, Incheon 21936, Korea.
Department of New Drug Development, Inha University College of Medicine, Incheon 22212, Korea.
Molecules. 2021 Jan 15;26(2):425. doi: 10.3390/molecules26020425.
Sirolimus is a hydrophobic macrolide compound that has been used for long-term immunosuppressive therapy, prevention of restenosis, and treatment of lymphangioleiomyomatosis. In this study, a simple and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed and validated for the simultaneous determination of sirolimus in both porcine whole blood and lung tissue. Blood and lung tissue homogenates were deproteinized with acetonitrile and injected into the LC-MS/MS system for analysis using the positive electrospray ionization mode. The drug was separated on a C18 reversed phase column with a gradient mobile phase (ammonium formate buffer (5 mM) with 0.1% formic acid and acetonitrile) at 0.2 mL/min. The selected reaction monitoring transitions of / 931.5 → 864.4 and 809.5 → 756.5 were applied for sirolimus and ascomycin (the internal standard, IS), respectively. The method was selective and linear over a concentration range of 0.5-50 ng/mL. The method was validated for sensitivity, accuracy, precision, extraction recovery, matrix effect, and stability in porcine whole blood and lung tissue homogenates, and all values were within acceptable ranges. The method was applied to a pharmacokinetic study to quantitate sirolimus levels in porcine blood and its distribution in lung tissue following the application of stents in the porcine coronary arteries. It enabled the quantification of sirolimus concentration until 2 and 14 days in blood and in lung tissue, respectively. This method would be appropriate for both routine porcine pharmacokinetic and bio-distribution studies of sirolimus formulations.
西罗莫司是一种疏水性大环内酯化合物，已被用于长期免疫抑制治疗、预防再狭窄和治疗淋巴管平滑肌瘤病。在这项研究中，开发并验证了一种简单而灵敏的液相色谱-串联质谱（LC-MS/MS）方法，用于同时测定猪全血和肺组织中的西罗莫司。用乙腈对血液和肺组织匀浆进行蛋白沉淀，然后注入 LC-MS/MS 系统，采用正电喷雾电离模式进行分析。药物在 C18 反相柱上以梯度流动相（5 mM 甲酸铵缓冲液含 0.1%甲酸和乙腈）在 0.2 mL/min 下进行分离。选择反应监测转换 / 931.5 → 864.4 和 809.5 → 756.5 分别用于西罗莫司和依维莫司（内标，IS）。该方法在 0.5-50 ng/mL 的浓度范围内具有选择性和线性。该方法已在猪全血和肺组织匀浆中进行了灵敏度、准确性、精密度、提取回收率、基质效应和稳定性验证，所有值均在可接受范围内。该方法应用于药代动力学研究，定量分析猪冠状动脉支架植入后猪血液中西罗莫司的浓度及其在肺组织中的分布。该方法能够定量检测血液中西罗莫司浓度直至 2 天和 14 天，肺组织中浓度直至 14 天。该方法适用于常规猪药代动力学和西罗莫司制剂的生物分布研究。
