RP-HPLC法测定野生刺五加果实中6种成分的含量

Determination of six components in wild Acanthopanax senticosus fruits by RP-HPLC

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RPHPLC方法。方法：采用Agilent Eclipse XDB-C18（4.6 mm×250 mm,5μm）色谱柱,以乙腈-0.3%磷酸水溶液为流动相,流速1.0 m L·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、2.522～18.915μg·m L-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=0.0799A＋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-C18column（ 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 m L·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 ·m L- 1,19. 224- 144. 18μg·m L- 1,18. 454-138. 405μg·m L- 1,8. 416- 63. 12μg·m L- 1,3. 286- 24. 645μg·m L- 1,and 2. 522- 18. 915 μg·m L- 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- 1and 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.