盐酸法舒地尔原料药有关物质HPLC法测定的研究

Investigation on the determination of HPLC related substances related to the API of fasudil hydrochloride

目的:建立盐酸法舒地尔原料药中法舒地尔吡啶N-氧化物、法舒地尔1位异构体、5-哌嗪盐酸法舒地尔等有关物质测定的HPLC法。方法:采用YMC-Pack Pro C_(18)色谱柱(4.6 mm×250 mm,5μm),以1.5%三乙胺(pH 5.4):乙腈=87∶13(A)-乙腈(B)为流动相,梯度洗脱,流速1.0 mL·min^(-1),柱温30℃,检测波长275 nm,进样量20μL。结果:盐酸法舒地尔中法舒地尔吡啶N-氧化物、法舒地尔1位异构体、5-哌嗪盐酸法舒地尔与主成分间均能有效分离,线性范围分别为0.2575~5.0590、0.2539~5.0780、0.2530~5.0590μg·mL^(-1),检测限为0.2058、0.1270、0.1265μg·mL^(-1),定量限为0.4115、0.4062、0.4047μg·mL^(-1),准确度、精密度、耐用性均符合要求。结论:该方法适用于盐酸法舒地尔原料药中法舒地尔吡啶N-氧化物、法舒地尔1位异构、5-哌嗪盐酸法舒地尔等有关物质的测定和质量控制。

Objective:To established a HPLC method for the determination of fasudil pyridine N-oxide,fasudil 1-isomer,5-piperazine hydrochloride and other related substances in fasudil hydrochloride API.Methods:The YMC-Pack Pro C_(18)column(4.6 mm×250 mm,5μm)was used with 1.5%triethylamine(pH 5.4):acetonitrile=87∶13(A)-acetonitrile(B)as the mobile phase,gradient elute,flow rate 1.0 mL·min^(-1),column temperature 30℃,detection wavelength 275 nm,injection volume 20μL.Results:Fasudil hydrochloride in fasudil hydrochloride pyridine N-oxide,fasudil 1 isomer,5-piperazine hydrochloride fasudil hydrochloride and the principal component can be effectively separated,the linear range were 0.2575-5.0590,0.2539-5.0780,0.2530-5.0590μg·mL^(-1),and the detection limits were 0.2058,0.1270,0.1265μg·mL^(-1),respectively.The quantitative limits were 0.4115,0.4062,0.4047μg·mL^(-1),and the accuracy,precision and durability meet the requirements.Conclusion:The method is suitable for the determination and quality control of fasudil pyridine N-oxide,fasudil 1-isomer,5-piperazine fasudil hydrochloride and other related substances in fasudil hydrochloride API.