RP-HPLC法测定野生刺五加果实中6种成分的含量

目的: 建立测定刺五加果实中原儿茶酸、紫丁香苷、绿原酸、刺五加苷E、异嗪皮啶和槲皮素-3-鼠李糖苷含量的RP-HPLC方法。方法: 采用Agilent Eclipse XDB-C₁₈(4.6 mm×250 mm,5 μm)色谱柱,以乙腈-0.3%磷酸水溶液为流动相,流速1.0 mL·min^-1,梯度洗脱,检测波长300 nm,柱温30 ℃。结果: 原儿茶酸、紫丁香苷、绿原酸、刺五加苷E、异嗪皮啶和槲皮素-3-鼠李糖苷质量浓度分别在4.92~36.9、19.224~144.18、18.454~138.405、8.416~63.12、3.286~24.645、2.522~18.915 μg·mL^-1范围内与峰面积呈良好的线性关系,其线性回归方程分别为C=0.055 2A-0.057 9(r=0.999 9),C=0.079 9A+0.014 6(r=0.999 7),C=0.012 7A-0.131 4(r=0.999 6),C=0.057 7A+0.086 6(r=0.999 8),C=0.012 0A-0.020 8(r=0.999 9),C=0.014 2A+0.065 1(r=0.999 9);加样回收率(n=6)分别为100.4%、99.8%、100.9%、101.2%、99.4%和99.7%。6个成分的含量测定结果分别为0.501~0.510、2.511~2.522、2.310~2.325、1.331~1.340、0.371~0.382和0.260~0.272 mg·g^-1。结论: 经方法学验证,该法可作为刺五加果实质量控制的方法。

Objective: To develop an RP-HPLC method for determination of protocatechuic acid, syringin, chlorogenic acid, eleutheroside E, isofraxidin, and quercetin-3-rhamnoside in Acanthopanax senticosus fruits.Methods: The determination was performed on an Agilent Eclipse XDB-C₁₈ column(4.6 mm×250 mm, 5 μm)at 30 ℃.The mobile phase was acetonitrile-0.3% aqueous phosphoric acid with gradient elution at a flow rate of 1.0 mL·min^-1.The detection wavelength was set at 300 nm.Results: The linear correlation equations for protocatechuic acid, syringin, chlorogenic acid, eleutheroside E, isofraxidin and quercetin-3-rhamnoside were C=0.055 2A-0.057 9(r=0.999 9), C=0.079 9A+0.014 6(r=0.999 7), C=0.012 7A-0.131 4(r=0.999 6), C=0.057 7A+0.086 6(r=0.999 8), C=0.012 0A-0.020 8(r=0.999 9), and C=0.014 2A+0.065 1(r=0.999 9), respectively.A good linearity was obtained over the range of 4.92-36.9μg·mL^-1, 19.224-144.18μg·mL^-1, 18.454-138.405μg·mL^-1, 8.416-63.12μg·mL^-1, 3.286-24.645μg·mL^-1, and 2.522-18.915 μg·mL^-1, respectively.The recoveries(n=6)were 100.4%, 99.8%, 100.9%, 101.2%, 99.4%, and 99.7%, respectively.The content of six components were in the ranges of 0.501-0.510 mg·g^-1, 2.511-2.522 mg·g^-1, 2.310-2.325 mg·g^-1, 1.331-1.340 mg·g^-1, 0.371-0.382 mg·g^-1 and 0.260-0.272 mg·g^-1, respectively.Conclusion: It is proved that the method can be used to control the quality of Acanthopanax senticosus fruits by verification methodology.