HPLC法同时测定草珊瑚中7个活性成分的含量

目的: 建立HPLC法同时测定草珊瑚药材中反丁烯二酸、新绿原酸、绿原酸、隐绿原酸、异嗪皮啶、落新妇苷、迷迭香酸含量。 方法: 采用Agilent Zorbax Eclipse XDB-C₁₈色谱柱(250 mm×4.6 mm,5 μm),流动相A为乙腈-甲醇(1: 9),流动相B为0.5%的甲酸水溶液,梯度洗脱(0~20 min,20%A;20~30 mim,20%A→30%A;30~60 min,30%A),流速为0.8mL·min^–1,检测波长为228 nm(检测反丁烯二酸)、344 nm(检测新绿原酸、绿原酸、隐绿原酸、异嗪皮啶、落新妇苷、迷迭香酸),柱温为40 ℃。 结果: 反丁烯二酸、新绿原酸、绿原酸、隐绿原酸、异嗪皮啶、落新妇苷、迷迭香酸浓度分别在24.12~180.90、1.01~101.00、1.75~175.04、2.19~219.20、1.27~127.20、1.57~157.08、2.93~193.28 μg·mL^-1范围内与峰面积呈良好的线性关系,平均加样回收率(n=6)均在93.92%~103.0%范围内,RSD均小于3.0%。 结论: 该方法准确,灵敏度高,重复性好,适用于草珊瑚药材中7个有效成分的含量测定。

Objective: To develop an HPLC method for simultaneous determination of seven main bioactive constituents including fumaric acid, neochlorogenic acid, chlorogenic acid, cryptochlorogenin acid, isofraxidin, astilbin, rosmarinic acid in the whole plant of Sarcandra glabra (Thunb.) Nakai. Methods: An Agilent Zorbax Eclipse XDB-C₁₈ column (250 mm×4.6 mm, 5 μm) was adopted and the column temperature was at 40 ℃. The mobile phase consisted of a water solution containing 0.5% formic acid and acetonitrile-methanol (1: 9) in gradient mode [0-20 min,20% acetonitrile-methanol(1: 9);20-30 mim, 20%→30% acetonitrile-methanol (1: 9);30-60 min,30% acetonitrile-methanol (1: 9)], the flow rate was 0.8 mL·min^-1, the detection wavelength was set at 228 nm for detecting fumaric acid and 344 nm for detecting neochlorogenic acid, chlorogenic acid, cryptochlorogenin acid, isofraxidin, astilbin, and rosmarinic acid. Results: The good linearity with peak areas was achieved in the range of 24.12-180.90 μg·mL^-1 for fumaric acid, 1.01-101.00 μg·mL^-1 for neochlorogenic acid, 1.75-175.04 μg·mL^-1 for chlorogenic acid, 2.19-219.20 μg·mL^-1 for cryptochlorogenin acid, 1.27-127.20 μg·mL^-1 for isofraxidin, 1.57-157.08 μg·mL^-1 for astilbin, 2.93-193.28 μg·mL^-1 for rosmarinic acid. The average recoveries(n=6) were 93.92%-103.0% with RSD less than 3.0%. Conclusion: The method developed is accurate, sensitive, reproducible, which can be used for the determination of the seven bioactive constituents in the whole plant of S. glabra.