Zhao Guang, Yuan Fusheng, Zhu Jiajun
Orthopaedic Department, the Fourth Affiliated Hospital of China Medical University, Shenyang, 110032, People's Republic of China.
Biomed Chromatogr. 2014 Mar;28(3):341-7. doi: 10.1002/bmc.3025. Epub 2013 Sep 2.
A rapid, simple, selective and sensitive LC-MS/MS method was developed for the determination of curculigoside in rat plasma. The analytical procedure involves extraction of curculigoside and syringin (internal standard, IS) from rat plasma with a one-step extraction method by protein precipitation. The chromatographic resolution was performed on an Agilent XDB-C18 column (4.6 × 50 mm, 5 µm) using an isocratic mobile phase of methanol with 0.1% formic acid and H2 O with 0.1% formic acid (45:55, v/v) at a flow rate of 0.35 mL/min with a total run time of 2.0 min. The assay was achieved under the multiple-reaction monitoring mode using positive electrospray ionization. Method validation was performed according to US Food and Drug Administration guidelines and the results met the acceptance criteria. The calibration curve was linear over 4.00-4000 ng/mL (R = 0.9984) for curculigoside with a lower limit of quantification of 4.00 ng/mL in rat plasma. The intra- and inter-day precisions and accuracies were 3.5-4.6 and 0.7-9.1%, in rat plasma, respectively. The validated LC-MS/MS method was successfully applied to a pharmacokinetic study of curculigoside in rats after a single intravenous and oral administration of 3.2 and 32 mg/kg. The absolute bioavailability of curculigoside after oral administration was 1.27%.
建立了一种快速、简便、选择性好且灵敏的液相色谱-串联质谱（LC-MS/MS）法，用于测定大鼠血浆中仙茅苷的含量。分析过程包括采用一步蛋白沉淀萃取法从大鼠血浆中萃取仙茅苷和紫丁香苷（内标，IS）。色谱分离在安捷伦XDB-C18柱（4.6×50 mm，5 µm）上进行，使用甲醇与0.1%甲酸和水与0.1%甲酸的等度流动相（45:55，v/v），流速为0.35 mL/min，总运行时间为2.0 min。采用正电喷雾电离在多反应监测模式下完成测定。按照美国食品药品监督管理局的指导原则进行方法验证，结果符合验收标准。仙茅苷的校准曲线在4.00 - 4000 ng/mL范围内呈线性（R = 0.9984），大鼠血浆中的定量下限为4.00 ng/mL。大鼠血浆中日内和日间精密度及准确度分别为3.5 - 4.6%和0.7 - 9.1%。经验证的LC-MS/MS方法成功应用于大鼠单次静脉注射和口服3.2和32 mg/kg仙茅苷后的药代动力学研究。口服给药后仙茅苷的绝对生物利用度为1.27%。
