盐酸法舒地尔中杂质和二聚体的HPLC法测定

Determination of impurities and dimers in fasudil hydrochloride by HPLC

目的采用高效液相色谱法建立盐酸法舒地尔中杂质检查和二聚体的测定。方法用Venusil XBP C18柱(250mm×4.6mm,5μm),甲醇–水(50∶50,含10mmol/L庚烷磺酸钠和5mmol/L磷酸二氢钠,用磷酸调pH3.0)为流动相;紫外检测波长为275nm;进样量10μL;体积流量1.0mL/min。采用自身对照法测定杂质,标准曲线法测定二聚体。结果盐酸法舒地尔中4个单一杂质量的均值不大于1.34%,之和的均值不大于1.38%。盐酸法舒地尔二聚体在0.63～41.60μg/mL线性关系良好,r=0.9999;平均回收率为99.5%,RSD值为1.3%。结论本方法简便、准确、专属性强,可作为盐酸法舒地尔中杂质检查及二聚体的测定方法。

Objective To develop an RP-HPLC method for determination of the impurities and dimers in fasudil hydrochloride. Methods Venusil XBP C 18 column （250 mm × 4.6 mm, 5 μm） was used. The mobile phase was composed of methanol-H 2 O （50∶50, including 10 mmol/L sodium heptane-1-sulphonate and 5 mmol/L sodium dihydrogen phosphate, pH 3.0）. The flow rate was 1.0 mL/min, and the determination wavelength was set at 275 nm with the injection volume of 10 μL. The total impurities were determined by the principal self contrast method, at the same time the dimmer contents in the sample were determined by standard curve method. Results Each and total contents of four impurities in fasudil hydrochloride were below 1.34% and 1.38%. A linearity between peak areas and dimer content was achieved in the range of 0.63—41.60 μg/mL（r = 0. 999 9）, and the average recovery was 99.5%, with an RSD value of 1.3%. Conclusion The method is simple, accurate, and sensitive, and could be used for the assay and quality control of impurities and dimers in fasudil hydrochloride.