HPLC法测定芪桂痛风片中新绿原酸、秦皮甲素、绿原酸、秦皮乙素、秦皮苷、秦皮素、4,5-O-二咖啡酰奎宁酸和蒙花苷

Determination of neochlorogenic acid,esculin,chlorogenic acid,esculetin,fraxin,fraxetin,4,5-O-dicaffeoylquinic acid,and linarin in Qigui Tongfeng Tablets by HPLC

目的 建立测定芪桂痛风片中新绿原酸、秦皮甲素、绿原酸、秦皮乙素、秦皮苷、秦皮素、4,5-O-二咖啡酰奎宁酸、蒙花苷的HPLC方法。方法 采用ACE Excel C18-AR色谱柱(150 mm×4.6 mm,3μm);流动相为0.1%甲酸水–乙腈,梯度洗脱;检测波长325 nm(新绿原酸、绿原酸、4,5-O-二咖啡酰奎宁酸)、334 nm(秦皮甲素、秦皮乙素、秦皮苷、秦皮素、蒙花苷);体积流量1 mL/min;柱温30℃;进样量10μL。结果 新绿原酸、秦皮甲素、绿原酸、秦皮乙素、秦皮苷、秦皮素、4,5-O-二咖啡酰奎宁酸、蒙花苷的线性范围分别为1.86~59.45、26.21~838.80、8.01~256.47、3.60~115.45、17.79~569.18、1.93~61.76、3.67~117.51、2.27~72.76μg/mL,线性关系良好(r=1.000 0),平均回收率分别为95.56%、92.88%、95.49%、92.15%、93.03%、96.03%、99.90%、95.42%,RSD值分别为3.40%、3.11%、3.25%、3.17%、3.10%、3.52%、3.40%、3.34%。结论 建立的芪桂痛风片中8种成分HPLC定量方法简便、快速、准确,可以用于评价芪桂痛风片的质量。

Objective To establish an HPLC method for determination of neochlorogenic acid, esculin, chlorogenic acid, esculetin,fraxin, fraxetin, 4,5-O-dicaffeoylquinic acid, and linarin in Qigui Tongfeng Tablets. Methods The separation was carried out on Ace Excel C18-AR column(150 mm × 4.6 mm,3 μm). The mobile phase was acetonitrile-0.1% formic acid with gradient elution. The detection wavelength was set at 325 nm(neochlorogenic acid, chlorogenic acid, and 4,5-O-dicaffeoylquinic acid) and 334 nm(esculin,esculetin, fraxin, fraxetin, and linarin). The flow rate was 1.0 mL/min, temperature of column was set at 30 ℃, and the volume of injection was 10 μL. Results Neochlorogenic acid, esculin, chlorogenic acid, esculetin, fraxin, fraxetin, 4,5-O-dicaffeoylquinic acid,and linarin had good linear relationships in concentration range of 1.86 — 59.45, 26.21 — 838.80, 8.01 — 256.47, 3.60 — 115.45,17.79 — 569.18, 1.93 — 61.76, 3.67 — 117.51, and 2.27 — 72.76 μg/mL(r=1.000 0), and their average recoveries were 95.56%,92.88%, 95.49%, 92.15%, 93.03%, 96.03%, 99.90%, and 95.42% with RSD values as 3.40%, 3.11%, 3.25%, 3.17%, 3.10%, 3.52%,3.40%, and 3.34%, respectively. Conclusion The established HPLC quantitative method for eight components in Qigui Tongfeng Tablets is simple, rapid, and accurate, and can be used to evaluate the quality of Qigui Tongfeng Tablets.