HPLC法同时测定复方利血平片中7个成分的含量

Simultaneous determination of the content of seven components in compound reserpine tablets by HPLC

目的:建立HPLC法同时测定复方利血平片中利血平、氢氯噻嗪、硫酸双肼屈嗪、盐酸异丙嗪、维生素B_(1)、维生素B_(6)和泛酸钙7个成分的含量。方法:采用Agilent Zorbax SB-C_(18)色谱柱(250 mm×4.6 mm,5μm),以乙腈(A)-0.03 mol·L^(-1)磷酸二氢钾溶液(含0.25%庚烷磺酸钠,用磷酸调节pH至4.0)(B)为流动相,梯度洗脱(0~7 min,5%A→_(18)%A;7~15 min,_(18)%A;15.1~23 min,50%A;23.1~30 min,5%A),流速1.0 mL·min^(-1),柱温40℃,检测波长205 nm(0~7 min,泛酸钙)、275 nm(7~17 min,氢氯噻嗪、维生素B_(6)、硫酸双肼屈嗪和维生素B_(1))、215 nm(17~30 min,盐酸异丙嗪和利血平),进样体积20μL。结果:利血平、氢氯噻嗪、硫酸双肼屈嗪、盐酸异丙嗪、维生素B_(1)、维生素B_(6)和泛酸钙的浓度在相应的范围内与峰面积呈良好线性关系(r均>0.999);低、中、高3个浓度水平9份样品的平均加样回收率分别为99.8%(RSD=1.4%)、98.9%(RSD=1.5%)、100.4%(RSD=1.4%)、99.8%(RSD=1.0%)、99.2%(RSD=1.2%)、100.9%(RSD=1.1%)、100.2%(RSD=1.2%);精密度、稳定性良好,RSD均<2.0%。薄膜衣片(6个生产企业共17个批次)中7个成分的含量均介于标示量的93.0%~108.2%;糖衣片(1个生产企业共3个批次)中7个成分的含量均介于标示量的75.2%~108.0%。结论:该方法简便快捷,专属性好,可用于复方利血平片的质量控制,为质量标准提高提供参考。

Objective:To establish an HPLC method for the simultaneous determination of reserpine,hydrochlorothiazide,dihydralazine sulfate,promethazine hydrochloride,vitamin B_(1),vitamin B_(6) and calcium pantothenate in compound reserpine tablets.Methods:HPLC experiment was performed on an column of Agilent Zorbax SB-C_(18)(250 mm×4.6 mm,5μm)with the mobile phase of acetonitrile(A)-0.03 mol·L^(-1) KH_(2)PO_(4)(containing0.25%of heptanesulfonic acid sodium and adjust pH to 4.0 with phosphoric acid)(B)with gradient elution(0-7 min,5%A→_(18)%A;7^(-1)5 min,_(18)%A;15.1-23 min,50%A;23.1-30 min,5%A).The flow rate was 1.0 mL·min^(-1),the column temperature was 40℃,and the detection wavelength was 205 nm(0-7 min for calcium pantothenate),275 nm(7^(-1)7 min for hydrochlorothiazide,vitamin B_(6),dihydralazine sulfate,vitamin B_(1))and 215 nm(17-30 min for promethazine hydrochloride and reserpine).The injection volume was 20μL.Results:Reserpine,hydrochlorothiazide,dihydralazine sulfate,promethazine hydrochloride,vitamin B_(1),vitamin B_(6) and calcium pantothenate showed good linear relationships with the peak area in the corresponding range(r>0.999),the average recoveries of 9 samples at low,medium and high concentration levels were 99.8%(RSD=1.4%),98.9%(RSD=1.5%),100.4%(RSD=1.4%),99.8%(RSD=1.0%),99.2%(RSD=1.2%),100.9%(RSD=1.1%),100.2%(RSD=1.2%)respectively.The precision and stability were good with RSDs less than 2%.The contents of 7 components in film-coated tablets(6 manufacturing enterprises,a total of 17 batches)were between 93.0%and 108.2%of the labeled amount.The contents of 7 ingredients in sugar-coated tablets were between 75.2%and 108.0%of the marked amount.Conclusion:The established method is simple,rapid,specific for the quality control of compound reserpine tablets.It provides reference for improving the quality standards of compound reserpine tablets.