HPLC-MS法测定家兔眼组织中环孢素含量

Determination of Cyclosporin A in Ocular Tissues by HPLC-MS

目的 :建立家兔眼部组织 :角膜 ,房水 ,虹膜 ,巩膜 ,晶体和玻璃体中环孢素的高效液相色谱 质谱测定方法。方法 :眼部液体组织或固体组织的匀浆液加入内标物环孢菌素D后经乙醚提取 ,挥干 ,残渣用 70 %甲醇水溶液复溶 ,再用正庚烷除去脂溶性杂质 ,进行高效液相色谱 质谱检测 ,色谱柱为SymmetryC18柱 (5 μm ,2 1mm× 15 0mm) ,流动相为甲醇 水 (90∶10 ) ,流速为 0 2 5ml/min ,质谱采用电喷雾电离源 ,选择性离子检测 (SIR) :环孢素m/z 12 2 5 [M +Na]+ ,内标物环孢菌素Dm/z 12 39[M +Na]+ 。结果 :不同眼组织中环孢素的回收率均在96 6 5 %～ 10 5 5 2 %之间 ,日内精密度均小于 7 6 3% ,日间精密度均小于 11 2 2 %。最低检测浓度可达 5ng/ml。结论 :本方法简便 ,专属 ,准确 ,灵敏度高 ,适用于环孢素眼部药代动力学及组织分布研究。

AIM:To establish a HPLC-MS method for the determination of cyclosporin A in ocular tissues of aqueous humor, vitreous humor, cornea, sclera, iris and lens. METHOD:Ocular tissue samples containing cyclosporin D as internal standard was extracted with ethyl ether. The high-performance liquid chromatography separation was accomplished on RP-C_ 18 column(2.1 mm×150 mm, 5 μm) with the mobile phase of methanol-water(90∶10) at flow rate of 0.25 ml/min. The quantification was carried out with the selected ion recording mode by monitoring the molecular ion CsA m/z 1225 [M+Na] + ,and CsD m/z 1239 [M+Na] + .RESULT:The recovery of method in different tissues ranged from 96.65% to 105.52%, the RSD of within-day was less than 7.63% and the RSD of between-day less than 11.86%. CONCLUSION:The method is specific, sensitive and rapid, which is suitable for determination of CsA in ocular tissues and for study on its ditribution and pharmacokinetics.