Bioanalysis of free and liposomal Amphotericin B in rat plasma using solid phase extraction and protein precipitation followed by LC‐MS/MS
Bioanalysis of free and liposomal Amphotericin B in rat plasma using solid phase extraction and protein precipitation followed by LC‐MS/MS
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DOI：
10.1016/j.jpba.2018.06.014
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发表时间：
2018-09
影响因子：
3.4
通讯作者：
Chong Su;Hong Yang;Hui Sun;J. P. Fawcett;Dong Sun;J. Gu
中科院分区：
文献类型：
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作者：
Chong Su;Hong Yang;Hui Sun;J. P. Fawcett;Dong Sun;J. Gu
两性霉素B（AMB）是一种多烯大环内酯类抗生素，用于治疗侵袭性真菌感染。脂质体AMB（L-AMB）是一种脂质剂型，可减少药物的副作用和毒性。体内游离AMB（F-AMB）和L-AMB的定量对于监测脂质体制剂的质量控制并确保其在临床使用期间的安全性是重要的。本研究采用LC-MS/MS法分别测定大鼠血浆中的F-AMB和L-AMB。F-AMB经固相萃取分离后进行分析，总AMB（T-AMB）经蛋白沉淀后测定，L-AMB经差分法测定。该方法得到充分验证。校准曲线在0.7-120 μg/mL（T-AMB）和0.2-20 μg/mL（F-AMB）范围内呈线性。准确度和精密度结果在可接受的变异性限度内，回收率一致且重现性好，基质效应不显著，分析物在所有检测的储存条件下均稳定。该方法成功地应用于大鼠单次静脉注射6 mg/kg剂量L-AMB的药代动力学研究。该方法将为L-AMB的进一步临床研究提供基础，并为其他脂质体药物制剂的含量测定提供有用的技术支持。
Amphotericin B (AMB) is a polyene macrolide antibiotic used for treating invasive fungal infections. Liposomal AMB (L-AMB) is a lipid dosage form which reduces the side effects and toxicity of the drug. The quantitation of free AMB (F-AMB) and L-AMB in vivo is important to monitor quality control of the liposomal formulation and to ensure its safety during clinical use. In this study, an original strategy was developed to separately determine F-AMB and L-AMB in rat plasma using LC–MS/MS. F-AMB was analyzed after separation by solid phase extraction, total AMB (T-AMB) was determined after protein precipitation and L-AMB was determined by difference. The method was fully validated. Calibration curves were linear in the ranges 0.7–120 μg/mL for T-AMB and 0.2–20 μg/mL for F-AMB. Accuracy and precision results were within acceptable variability limits, recoveries were consistent and reproducible, matrix effects were insignificant and analytes were stable under all the storage conditions tested. The method was successfully applied to a pharmacokinetic study in rats administered a single intravenous 6 mg/kg dose of L-AMB. The method will allow further clinical studies of L-AMB and provide useful technical support for the assay of other liposomal drug formulations.