高效液相色谱-二极管阵列检测-电化学检测联用技术同时测定三精双黄连口服液中的4种化合物

Simultaneous Determination of Four Compounds in Sanjing Shuang-huanglian Oral Liquid by High Performance Liquid Chroma-tography-Diode Array Detection-Electrochemical Detection

建立了应用高效液相色谱-二极管阵列检测-电化学检测（HPLC—DAD—ECD）联用技术同时测定三精双黄连口服液中的绿原酸、咖啡酸、黄芩甙和木樨草素的方法。以Zorbax SB—C18柱（150mm×4．6mmi．d．，5．0μm）为色谱柱，柱温为30℃，流动相为（A）甲醇和（B）甲醇-水-冰醋酸（体积比为50：50：1），其梯度洗脱程序为2％A3min↑→ 3%A12min → 25%A 5min↑→80%A。 流速为0．8mL／min。二极管阵列检测波长为275nm。电化学单安培检测器的工作电压为0．7V。在上述条件下实现了绿原酸、咖啡酸、黄芩甙和木樨草素的分离检测。上述4种化合物的回收率为96．6％～99．6％，相对标准偏差（RSD）为2．5％～4．1％，检测限依次为1，0．2，9和7mg／L。该方法简便、快速，重现性和准确度较好，可作为测定双黄连口服液中绿原酸、咖啡酸、黄芩甙和木樨草素的有效方法。

Chlorogenic acid, caffeic acid, baicalin and luteolin in Sanjing Shuanghuanglian Oral Liquid were simultaneously detected and identified using a high performance liquid chromatography coupled with diode array detection and electrochemical detection （HPLC-DAD-ECD）. The separation was performed on a Zorbax SB-C18 column （ 150 mm×4.6 mm i. d. , 5.0 μm）. The mobile phase consisted of （A） methanol and （B） methanol-water-acetic acid （50： 50： 1, v/v/v） using a linear gradient elution of 2%A -3%A at 0 -3 min, 3%A -25%A at 3 - 15 min, 25%A - 80%A at 15 - 20 min. The flow rate was 0.8 mL/min. The DAD detection was used at 275 nm. The ECD detection was done at 0. 7 V. The column thermostat set at 30 ℃. The limits of detection of the 4 compounds were 1 mg/L for chlorogenic acid, 0.2 mg/L for caffeic acid, 9 mg/L for baicalin, 7 mg/L for luteolin. The average recoveries were between 96.6% - 99.6% with relative standard deviations （RSDs） of 2.5% -4. 1%. The method is simple, rapid, reproducible and accurate. It can be used for the routine analysis of the four compounds in Shuanghuanglian Oral Liquid.