HPLC法测定疏风清热合剂中黄芩苷的含量

Determination of Baicalin in Shufeng Qingre Mixture by HPLC

目的:建立HPLC法测定疏风清热合剂中黄芩苷含量的方法。方法:采用HPLC法,色谱柱:Kromasil C18柱(250mm×4.6mm,5μm);流动相:甲醇-水-磷酸(47:53:0.2);流速:1.00mL/min;检测波长280nm;分别精密吸取对照品溶液、供试品溶液和阴性对照溶液,利用上述色谱条件,取20μL注入液相色谱仪,记录色谱图。结果:(1)从色谱图上看出,在该色谱条件下,保留时间约为4.1 min处,供试品和对照品溶液出现色谱峰,而阴性对照溶液未见此色谱峰,说明黄芩苷的测定不受其它杂质峰的干扰;(2)黄芩苷在1.98~39.56μg/mL浓度范围内线性关系良好(r=0.9998);(3)平均回收率97.3%,RSD=0.67%;(4)稳定性:黄芩苷峰面积的RSD=0.72%,表明供试品溶液在12 h内稳定性良好;(5)重复性:黄芩苷含量的RSD=0.61%,表明重现性较好;(6)黄芩苷含量:批号20180104黄芩苷含量0.63 mg/mL,RSD=0.68%,批号20180303黄芩苷含量0.51 mg/mL,RSD0.59%,批号20180503黄芩苷含量0.58 mg/mL,RSD0.77%。结论:HPLC法测定疏风清热合剂中黄芩苷含量方法简便、快速、准确,专属性强,可作为疏风清热合剂中黄芩苷的含量测定方法。

Objective:To establish the HPLC method for determination of Baicalin in Shufeng Qingre Mixture.Methods:HPLC method was adopted,chromatographic column:Kromasil C18 column(250 mm×4.6 mm,5 m),mobile phase:methanol-water-phosphoric acid(47:53:0.2),flow rate:1.00 mL/min,detection wavelength 280 nm.The chromatogram was recorded by injecting 20 mL into the liquid chromatograph under the above chromatographic conditions.Results:(1)The chromatogram showed that the chromatographic peak appeared at the retention time of about 4.1 min,but not in the negative control solution,indicating that the determination of baicalin was not interfered by other impurity peaks.(2)Baicalin was linear in the concentration range of 1.98-39.56 g/mL(r=0.9998).(3)The average recovery was 97.3%,RSD=0.67%.(4)Stability:The baicalin peak area RSD was 0.72%,indicating that the sample solution was stable at 12 h.(5)Reproducibility:The baicalin content RSD was 0.61%with good reproducibility.(6)Baicalin content:The baicalin content of batch number 20180104 was 0.63 mg/mL,RSD=0.68%,batch number 201803 was 0.51 mg/mL,RSD 0.59%,and batch number 201803 was 0.58 mg/mL,RSD 0.77%.Conclusion:HPLC method for determination of Baicalin in Shufeng Qingre Mixture is simple,rapid,accurate and specific.It can be used as a method for determination of Baicalin in Shufeng Qingre Mixture.