LC-MS/MS法测定人全血中环孢素A浓度及环孢素眼用乳剂健康人体药代动力学研究

建立液相色谱 质谱联用法测定人全血中环孢素A浓度，选用Kromasil-C₁₈色谱柱（20 mm×4.6 mm×5 μm），以甲醇1 mmol•L^-1甲酸铵溶液为流动相，采用梯度洗脱进行分离，样品用沉淀蛋白法处理后进样，流速1.1 mL•min^-1，柱温60 ℃，进样量20 μL。选用3200 QTrap型三重四极杆串联质谱仪的多重反应监测（MRM）扫描方式进行检测。环孢素A的线性范围为0.2~20.00 μg•L^-1，定量下限为0.2 μg•L^-1。准确度与精密度结果显示，方法日间、日内变异均小于15%，相对偏差为-12.60%~12.80%，方法提取回收率为（98.1±4.7）%，稳定性较好。所建立的方法快速、灵敏、专属性强、重现性好，可用于环孢素眼用乳剂健康人体药代动力学研究。

LC-MS/MS Determination of Cyclosporin in Human Blood and Application in Pharmacokinetics

The Cyclosporin in human blood was determinated by LC-MS/MS. Cyclosporin and the internal standard were extracted from blood by methanol, which was used as deproteinated solvent, and then separated on a Kromasil-C_(18) column (20 mm×4.6 mm×5 μm). The mobile phase was consisted of methanol-1 mmol·L~(-1) ammonium formate maintained at 60 ℃. The flow rate was 1.1 mL·min~(-1), and 20 μL aliquot of residues were injected into the LC-MS/MS system. Detection was carried out by multiple reaction monitoring on 3200 Qtrap LC-MS/MS system. The assay is linear over the range of 0.20-20.00 μg·L~(-1) with a lower limit of quantitation of 0.20 μg·L~(-1). Intra- and inter-day precision are less than 15%. The relative deviation is in the range of -12.60%-12.80%. The recovery of Cyclosporin is (98.1±4.7)%, and stability is good. It is a rapid, sensitive, selective and reliable method for the determination of Cyclosporin in human blood. The assay can be applied for the determination of Cyclosporin in human blood and the study on pharmacokinetics.