HPLC法测定注射用黄芩苷原料中黄芩苷及其有关物质的含量

Content Determination of Baicalin and Its Related Substances in Baicalin for Injection by HPLC

目的:建立测定注射用黄芩苷原料中黄芩苷及其有关物质含量的方法。方法:以高效液相色谱法测定注射用黄芩苷原料中黄芩苷及其有关物质千层纸素A-7-O葡糖醛酸苷、汉黄芩苷的含量,并以不加校正因子的主成分自身对照法计算总杂质的含量。色谱柱为Agilent ZORBAX SB-C18(150 mm×4.6 mm,5μm),流动相为甲醇-0.1%磷酸水溶液(梯度洗脱),流速为1.0 ml/min,柱温为30℃,检测波长为274 nm。结果:黄芩苷、千层纸素A-7-O葡糖醛酸苷、汉黄芩苷的质量浓度分别在50.00~300.00、0.21~6.57、0.16~0.80μg/ml范围内与各自峰面积积分值呈良好的线性关系(r分别为0.999 9、0.999 3、0.999 3);平均加样回收率分别为99.16%、100.69%、99.18%,RSD分别为1.2%、2.4%、4.6%(n均为9)。黄芩苷与有关物质分离良好,千层纸素A-7-O葡糖醛酸苷与汉黄芩苷的检出限分别为0.43、0.64 ng。结论:该方法专属性强,结果准确、可靠,可用于注射用黄芩苷原料中黄芩苷及其有关物质的含量测定。

OBJECTIVE： To establish a method for the content determination of baicalin and related substances in Baicalin for injection. METHODS： The contents of baicalin and related substance, such as oroxylin A 7-O-glucuronide, wogonoside, were de- termined by HPLC. The contents of the total impurities were computed by main constituent self-control method without calibration factor. The separation was performed on Agilent ZORBAX SB-C,8（ 150 mm^4.6 ram, 5 p.m） column with mobile phase consisted of methanol-0.1%phosphoric acid solution （gradient elution） at the flow rate of 1.0 ml/min. The column temperature was 30 ~C and the detection wavelength was set at 274 nm. RESULTS： The linear ranges of baicalin, oroxylin A 7-O-glucuronide and wogonoside were 50.00-300.00 pg/ml（r=0.999 9）,0.21-6.57 ~tg/ml（r----0.999 3） and 0.16-0.80 p.g/ml（r=0.999 3）. Average recovery rates were 99.16% （RSD= 1.2%, n----9）, 100.69% （RSD--2.4%, n----9） and 99.18% （RSD=4.6%, n-~9）. Baicalin was separated from related substance well. The detection limits of oroxylin A 7-O-glucureonide and wogonside were 0.43,0.64 rig. CONCLUSIONS： The meth- od is specific, accurate and reliable, and it can be used for the content determination of baicalin and related substance in baicalin for injection.