HPLC法同时测定炒牛蒡子饮片中7个活性成分的含量

目的：建立HPLC法同时测定炒牛蒡子饮片中绿原酸、隐绿原酸、3，4-二咖啡酰奎宁酸、3，5-二咖啡酰奎宁酸、4，5-二咖啡酰奎宁酸、牛蒡苷、牛蒡苷元7个主要活性成分的含量。方法：采用YMC-Pack-ODS-A C₁₈ （250 mm×4.6 mm，5 μm）色谱柱，流动相A为0.1%甲酸水溶液，流动相B为乙腈，梯度洗脱（0~20 min，5% B→15% B；20~75 min，15% B→35% B；75~90 min，35% B→50% B），流速为1.0 mL·min^-1，检测波长为286 nm，柱温为35℃。结果：绿原酸、隐绿原酸、3，4-二咖啡酰奎宁酸、3，5-二咖啡酰奎宁酸、4，5-二咖啡酰奎宁酸、牛蒡苷、牛蒡苷元质量浓度分别在12.55~200.80、17.84~89.20、2.44~122.00、10.26~513.00、5.48~274.00、44.63~1 428.00、12.83~205.20 μg·mL^-1范围内与峰面积呈良好的线性关系（r=0.999 3~1.000），平均加样回收率（n=6）均在94.3%~100.8%范围内，RSD均小于3.0%。样品中7个成分的含量测定结果分别为1.99~6.21、1.16~1.75、1.35~2.18、1.14~2.96、1.77~4.98、34.81~74.26、2.38~9.19 mg·g^-1。结论：该方法可以用于炒牛蒡子饮片中7个主要活性成分的含量测定。

Objective:To develop an HPLC method for simultaneous determination of seven main bioactive constituents (chlorogenic acid,cryptochlorogenic acid,3,4-dicaffeoylquinic acid,3,5-dicaffeoylquinic acid,4,5-dicaffeoylquinic acid,arctiin and arctigenin) in processed Fructus Arctii.Methods:A YMC-Pack-ODS-A C₁₈ column (250 mm×4.6 mm,5 μm) was used and the column temperature was 35. The mobile phase consisted of 0.1% formic acid (A) and acetonitrile (B) with elution gradient (5%B→15%B at 0-20 min,15%B→35%B at 20-75 min,35%B→50%B at 75-90 min) at a flow rate of 1 mL·min^-1. The detection wavelength was set at 286 nm. Results:The calibration curves of chlorogenic acid,cryptochlorogenic acid,3,4-dicaffeoylquinic acid,3,5-dicaffeoylquinic acid,4,5-dicaffeoylquinic acid,arctiin and arctigenin were in good linearity within 12.55-200.80,17.84-89.20,2.44-122.00,10.26-513.00,5.48-274.00,44.63-1 428.00 and 12.83-205.20 μg·mL^-1 (r=0.999 3-1.000),respectively. The average recoveries (n=6)were in the ranges of 94.3%-100.8% with RSD less than 3.0%.The contents in 12 batches of processed Fructus Arctii were 1.99-6.21 mg·g^-1 for chlorogenic acid,1.16-1.75 mg·g^-1 for cryptochlorogenic acid,1.35-2.18 mg·g^-1 for 3,4-dicaffeoylquinic acid,1.14-2.96 mg·g^-1 for 3,5-dicaffeoylquinic acid,1.77-4.98 mg·g^-1 for 4,5-dicaffeoylquinic acid,34.81-74.26 mg·g^-1 for arctiin and 2.38-9.19 mg·g^-1 for arctigenin. Conclusion:The method developed can be used for the determination of the seven bioactive constituents in processed Fructus Arctii.