结膜下给药兔眼玻璃体内维拉帕米的高效液相色谱分析

Determination of the concentration of verapamil in vitreous following subconjunctival administration by HPLC

目的 建立反相高效液相色谱法(HPLC)测定兔眼玻璃体内维拉帕米的浓度。方法 制备外伤性增生性玻璃体视网膜病变(PVR)动物模型,结膜下注射125mg/05ml维拉帕米,按时抽取玻璃体样品,经提取、浓缩处理后,采用HP1050HPLC系统测定样品中维拉帕米的浓度。结果 玻璃体内维拉帕米浓度在01～1000μg/ml范围内呈线性关系,回归方程为y=05933x,r=09992;变异系数为3%。最低检测浓度为005μg/ml,回收率大于95%。结论 此方法回收率高,线性关系良好,变异系数小,稳定可行,可推广应用。

ObjectiveTo develop a reversed phase high performance liquid chromatographic(HPLC) method for determining the concentration of verapamil in vitreous following subconjunctival administration.MethodsAn experimental model of traumatic proliferative vitreoretinopathy(PVR) was produced in pigment rabbits,which received subconjunctival verapamil injection after surgery.Vitreous specimens,obtained at various time,as well as the internal standard,imipramine,were extracted from alkalinised vitreous with n hexane isobutyl alcohol,then back extracted into 0 01*!mol/L phosphoric acid and the concentration of verapamil was determined by a reversed phase column and ultraviolet monitoring at 278*!nm.ResultsThe calibrated concentration curves of verapamil was linear with the concentration range of 0 1～100 0*!μg/ml( y=0 593*!3x,r=0 999*!2 ),the variation of average coefficient is 3%.The detection limit of verapamil is below 0 05*!μg/ml.Conclusion This HPLC method is specific,sensitive for detection the concentration of verapamil.Verapamil penetrated into the vitreous following subconjunctival administration and reached potentially therapeutic concentrations in 24*!h.