HPLC法测定复方利血平片中6个成分的含量均匀度

HPLC determination for the content uniformity of the six components in compound reserpine tablets

目的:建立HPLC法测定复方利血平片中氢氯噻嗪、硫酸双肼屈嗪、维生素B1、维生素B6、盐酸异丙嗪及利血平的含量均匀度。方法:采用Phenomenex Gemini C18色谱柱(250 mm×4.6 mm,5μm),柱温40℃,流速1.0 m L·min-1,以0.14%庚烷磺酸钠溶液(用5%磷酸溶液调节pH至3.5)-乙腈(82∶18)为流动相(1),检测波长为256 nm,测定氢氯噻嗪、硫酸双肼屈嗪、维生素B1和维生素B6;以0.14%庚烷磺酸钠溶液(用5%磷酸溶液调节pH至3.5)-乙腈(60∶40)为流动相(2),检测波长为220 nm,测定盐酸异丙嗪和利血平。结果:氢氯噻嗪、硫酸双肼屈嗪、维生素B1、维生素B6、盐酸异丙嗪及利血平含量均匀度的平均值(A+1.8S)分别为100.5%~103.4%(2.7~5.7),101.8%~105.3%(5.6~8.4),102.8%~104.8%(8.2~9.6),97.6%~99.8%(2.8~6.7),102.5%~103.6%(5.4~9.9),97.5%~%(12.2~14.1);定量限分别为1.6、2.7、7.8、26.2、1.7、0.7 ng;线性范围分别为0.036 88~0.086 04 mg·mL-1103.4(r=0.999 9), 0.050 95~0.118 9 mg·mL-1(r=0.999 6),0.011 69~0.027 27 mg·mL-1(r=0.999 4), 0.013 10~0.030 58 mg·mL-1(r=0.999 6),0.082 65~0.289 3 mg·mL-1(r=1.000),1.221~4.273μg·mL-1(r=1.000);低、中、高3个浓度共9份样品的平均回收率分别为99.9%(RSD为0.4%)、100.7%(RSD为1.4%)、100.5%(RSD为0.3%)、101.2%(RSD为0.5%)、100.6%(RSD为0.5%)和100.5%(RSD为0.7%),可同时鉴别氢氯噻嗪、硫酸双肼屈嗪、维生素B1、维生素B6、盐酸异丙嗪及利血平。结论:经方法学验证,该方法可用于复方利血平片的质量控制,且方法简便快捷,专属性强,重复性好。

Objective：To establish an HPLC method for the determination of the content uniformity of the six components（hydrochlorothiazide,dihydralae,vitamin B1,vitamin B6,promethazine hydrochloride and reserpine）in compound reserprine tablets.Mzienteh osduslfat：The Phenomenex Gemini C18 column（250 mm×4.6 mm,5 μm）was adopted at the flow rate of 1.0 mL ·min-1 with the column temperature of 40 ℃.The mobile phase（1）was 0.14% sodium 1-heptanesulfonate solution（adjust p H to 3.5 with 5% phosphoric acid solution）-acetonitrile（82∶18）and the detection wavelength was 256 nm for hydrochlorothiazide,dihydralazine sulfate,vitamin B1 and vitamin B6.The mobile phase（2）was 0.14% sodium 1-heptanesulfonate solution（adjust p H to 3.5 with 5% phosphoric acid solution）-acetonitrile（60∶40）,and the detection wavelength was 220 nm for promethazine hydrochloride and reserpine.Results：The averages（A＋1.8 S）of the content uniformity for the six components（hydrochlorothiazide,dihydralazinesulfate,vitamin B1,vitamin B6,promethazine hydrochloride and reserpine）were 100.5%-103.4%（2.7-5.7）,101.8%-105.3%（5.6-8.4）,102.8%-104.8%（8.2-9.6）,97.6%-9.8%（2.8-6.7）,102.5%-103.6%（5.4-9.9）,97.5%-103.4%（12.2-14.1）.The limits of quantitation of the six components were 1.6,2.7,7.8,26.2,1.7 and 0.7 ng,respectively.The linear ranges of the six components were 0.036 88-0.086 04 mg·m L-1（r=0.999 9）,0.050 95-0.118 9 mg·m L-1（r=0.999 6）,0.011 69-0.027 27 mg·m L-1（r=0.999 4）,0.013 10-0.030 58 mg·mL-1（r=0.999 6）,0.082 65-0.289 3 mg·mL-1（r=1.000）,1.221-4.273 μg·mL-1（r=1.000）.The average recoveries（n=9）from three concentrations of low,medium and high samples for the six components were 99.9%（RSD=0.4%）,100.7%（RSD=1.4%）,100.5%（RSD=0.3%）,101.2%（RSD=0.5%）,100.6%（RSD=0.5%）and 100.5%（RSD=0.7%）,respectively.Hydrochlorothiazide,dihydralazine sulfate,vitamin B1,vitamin B6,promethazine hydrochloride and reserpine were indentified simultaneously in the method.Conclusion：The method is simple,rapid,reproducible and specific for the quality control of compound reserprine tablets after validation.