高效液相色谱法测定盐酸黄酮哌酯原料和制剂中有关物质

摘要

Objective To establish an HPLC method for determination of the related substances in active pharmaceutical ingredient ( API ) and preparations of flavoxate hydrochloride, and to investigate the real impurity levels of the products in 6 manufacturers. Methods The Phenomenex Luna C18 column ( 150 mm × 4. 6 mm, 5 μm ) was adopted, the mobile phase was buffer solution ( taken 1 g sodium hexane sulfonate and added water to make it dissolve, added 1 mL three ethylamine and 1 mL phosphoric acid, and then added water to 650 mL, shaken well ) -acetonitrile ( 65:35 ) , gradient elution, the flow rate was 1. 0 mL/min, the detection wavelength was 293 nm, and the column temperature was 30 ℃. Results The peak occurrence sequence was as follows:flavoxate hydrochloride, impurity A, impurity B, impurity C and impurity D, the separation degree of each peak was more than 1. 5. The linear ranges of impurity A, impurity B, impurity C, impurity D and flavoxate hydrochloride were 0. 2922-5. 8440, 0. 3063-6. 1260, 0. 3087-6. 1738, 0. 3212-6. 4232, 0. 1081-2. 1620 μg/mL. The average recoveries of impurities A, B, C and D were 98. 92％, 99. 11％, 100. 04％, 99. 48％, RSDs were 0. 62％, 1. 34％, 1. 02％, 1. 43％ ( n=6 ) , respectively, and the correction factors were 1. 5, 1. 4, 1. 3 and 1. 2, respectively. Conclusion The method has high sensitivity and good separation effect, it can simultaneously determine several process impurities and degradation impurities in flavoxate hydrochloride.%目的 建立测定盐酸黄酮哌酯原料及制剂中有关物质的高效液相色谱(HPLC)法,考察6个厂家产品的真实杂质水平.方法 色谱柱采用Phenomenex Luna C18柱(150 mm×4.6 mm,5μm),流动相为缓冲液(取1 g己烷磺酸钠,加水适量使溶解,加三乙胺1 mL和磷酸1 mL,加水稀释至650 mL,摇匀)-乙腈(65:35),梯度洗脱,流速为1.0 mL/min,检测波长为293 nm,柱温为30℃.结果 盐酸黄酮哌酯、杂质A、杂质B、杂质C、杂质D依次出峰,各峰间分离度均大于1.5;杂质A,B,C,D及盐酸黄酮哌酯质量浓度分别在0.2922～5.8440,0.3063～6.1260,0.3087～6.1738,0.3212～6.4232,0.1081～2.1620μg/mL范围内与峰面积线性关系良好;杂质A,B,C,D平均回收率分别为98.92％,99.11％,100.04％,99.48％,RSD分别为0.62％,1.34％,1.02％,1.43％(n=6),校正因子分别为1.5,1.4,1.3,1.2.结论 该方法灵敏度高,分离效果好,能同时测定盐酸黄酮哌酯中几种工艺杂质和降解杂质.