高效液相色谱法同时测定复方罗布麻片I中7个成分的含量

目的：应用高效液相色谱技术，建立同时测定复方罗布麻片I中绿原酸、氢氯噻嗪、金丝桃苷、防己诺林碱、粉防己碱、蒙花苷和盐酸异丙嗪含量的方法。方法：采用Waters XBridge C₁₈色谱柱（250 mm×4.6 mm，5 μm），流动相为乙腈-0.2%磷酸水溶液，梯度洗脱，流速0.5 mL·min^-1，柱温30℃，检测波长为210、256、325 nm。结果：7个化学成分分离度良好，绿原酸、氢氯噻嗪、金丝桃苷、防己诺林碱、粉防己碱、蒙花苷、盐酸异丙嗪质量浓度分别在1.814~45.36 μg·mL^-1（r=1.000）、48.64~1 216 μg·mL^-1（r=0.999 8）、4.14~103.5 μg·mL^-1（r=1.000）、3.338~83.46 μg·mL^-1（r=1.000）、6.68~167 μg·mL^-1（r=1.000）、0.710 4~17.76μg·mL^-1（r=1.000）和3.128~78.2 μg·mL^-1（r=1.000）与峰面积呈良好的线性关系；精密度良好，相对标准偏差（RSD）小于0.90%；回收率良好，3个加样浓度的平均回收率（n=3）为98.5%~101.0%，RSD为0.38%~1.8%；稳定性良好，7个成分在24 h内保持稳定，RSD小于0.80%。8批样品中绿原酸、氢氯噻嗪、金丝桃苷、防己诺林碱、粉防己碱、蒙花苷、盐酸异丙嗪的含量测定结果分别为0.056 88~0.237 5、1.447~1.582、0.065 48~0.147 7、0.055 33~0.103 7、0.124 2~0.233 1、0.013 74~0.045 01和0.916 0~1.041 mg·片^-1。结论：该方法经方法学验证，可同时测定复方罗布麻片I中7个化学成分的含量，可用于该药品的质量控制。

Objective: To develop a high-performance liquid chromatography(HPLC) method for simultaneous determination of chlorogenic acid,hydrochlorothiazide,hyperoside,fangchinoline,tetrandrine,buddleoside and promethazine hydrochloride in Compound Kendir Leaves tablets I.Methods: A Waters XBridge C₁₈ column(250 mm×4.6 mm,5 μm) was used for the separation,with gradient elution of acetonitrile and 0.2% phosphoric acid as the mobile phase at a flow rate of 0.5 mL·min^-1. The detection wavelength was set at 210,256 and 325 nm. The column temperature was 30℃.Results: The seven constituents were well separated.The linear range was 1.814-45.36 μg·mL^-1 for chlorogenic acid(r=1.000),48.64-1216 μg·mL^-1 for hydrochlorothiazide(r=0.999 8),4.14-103.5 μg·mL^-1 for hyperoside(r=1.000),3.338-83.46 μg·mL^-1 for fangchinoline(r=1.000), 6.68-167 μg·mL^-1 for tetrandrine(r=1.000),0.7104-17.76 μg·mL^-1 for buddleoside(r=1.000),and 3.128-78.2 μg·mL^-1 for promethazine hydrochloride(r=1.000).The results of the precision tests were satisfactory with relative standard deviations(RSDs) all below 0.90%;the average spike recoveries(n=3) at three concentration levels were 98.5%-101.0% and RSDs were 0.38%-1.8%.The results of the stability tests were satisfactory and the seven constituents remained stable in 24 hours with all RSDs below 0.80%.The contents of chlorogenic acid,hydrochlorothiazide,hyperoside,fangchinoline,tetrandrine,buddleoside and promethazine hydrochloride in eight batches of Compound Kendir Leaves tablets I were 0.056 88-0.115 8,1.447-1.582,0.075 48-0.147 7, 0.055 33-0.103 7,0.124 2-0.233 1,0.013 74-0.045 01 and 0.916 0-1.041 mg per tablet,respectively.Conclusion: It is proved by methodology validation that the established method can be utilized for simultaneous determination of seven bioactive constituents in Compound Kendir Leaves tablets I,which can be used for the quality control of this drug.