HPLC-UV-ELSD联用同时测定痰热清注射液中6种成分的含量

目的:建立HPLC-UV-ELSD联用法同时测定痰热清注射液中6种有效成分(原儿茶酸,绿原酸,咖啡酸,黄芩苷,熊去氧胆酸,鹅去氧胆酸)的含量。方法:在相同色谱条件下,采用不同检测器同时测定痰热清注射液中6种有效成分,HPLC-UV法测定原儿茶酸、绿原酸、咖啡酸、黄芩苷,HPLC-ELSD测定熊去氧胆酸、鹅去氧胆酸。采用Venusil XBP-C₁₈色谱柱(4.6 mm×250 mm,5 μm),以乙腈(A)-4%醋酸水(B)为流动相,线性梯度洗脱,流速为1 mL·min^-1,紫外检测波长300 nm,ELSD漂移管温度70℃。结果:原儿茶酸、绿原酸、咖啡酸、黄芩苷、熊去氧胆酸、鹅去氧胆酸质量浓度分别在0.02~0.42 μg·mL^-1(r=0.9996)、0.31~6.12 μg·mL^-1(r=0.9993)、0.59~11.84 μg·mL^-1(r=0.9991)、33.24~664.75 μg·mL^-1(r=0.9996)、32.02~640.50 μg·mL^-1(r=0.9994)和5.58~111.60 μg·mL^-1(r=0.9998)范围内呈良好的线性关系;平均回收率在97.8%和101.5%之间。结论:该方法专属性强,耐用性好,可用于痰热清注射液中6种成分的含量测定和质量控制。

Objective:To establish a method for simultaneous determination of six active ingredients[protocatechuic acid,chlorogenic acid,caffeic acid,baicalin,ursodeoxycholic acid(UDCA)and chenodeoxycholic acid(CDCA)] in Tanreqing injection.Methods:The analysis and detection of six active ingredients in Tanreqing injection were carried out simultaneously under the same chromatographic condition.Protocatechuic acid,chlorogenic acid,caffeic acid and baicalin were detected by UV detector, and ursodeoxycholic acid(UDCA)and chenodeoxycholic acid(CDCA)were detected by ELSD detector.The determination was carried out on a Venusil XBP-C₁₈ column(4.6 mm×250 mm,5 μm)with gradient elution using acetonitrile(A)-4% aqueous ethylic acid(B)as mobile phase,and the flow rate was 1.0 mL·min^-1.The UV detection wavelength was 300 nm and the drift tube temperature of the evaporative light scattering detector (ELSD)was maintained at 70℃.Results:The calibration curves were linear in the ranges of 0.02-0.42 μg·mL^-1(r=0.9996)for protocatechuic acid,0.31-6.12 μg·mL^-1(r=0.999 3)for chlorogenic acid,0.59-11.84 μg·mL^-1(r=0.9991)for caffeic acid,33.24-664.75 μg·mL^-1(r=0.9996)for baicalin,32.02-640.50 μg·mL^-1(r=0.9994)for ursodeoxycholic acid,5.58-111.60 μg·mL^-1(r=0.9998)for chenodeoxycholic acid. The average recoveries fell between 97.8% and 101.5%.Conclusion:The established method can be used to determine the contents of protocatechuic acid,chlorogenic acid,caffeic acid,baicalin,ursodeoxycholic acid and chenodeoxycholic acid in Tanreqing injection,and thus may also serve as a standard for the quality control of Tanreqing injection.