HPLC测定覆盆子中鞣花酸、黄酮和覆盆子苷-F5的含量

Determination of ellagic acid,flavonoids and goshonoside-F5 in Rubi Fructus by HPLC

该研究建立了HPLC-uV同时测定覆盆子药材中鞣花酸、异槲皮苷、山柰酚-3-O-β-D-芸香糖苷、椴树苷和山柰酚含量以及HPLC-ELSD测定覆盆子苷-F5含量的方法。实验结果表明，各化合物的标准曲线线性关系良好（R^2〉0．9995），检测限为0．297～90．144ng，定量限为0．990～300．480ng，加样回收率为97．11％～101．7％（RSD≤2．1％）。不同产地覆盆子药材中各化合物的含量差异较大，其中鞣花酸的产量与纬度呈正相关，总黄酮含量在东部产地药材中较高，且覆盆子苷-F5含量在安徽亳州药材中最高。该研究建立的方法简便、准确、灵敏、重复性好，为覆盆子药材的进一步研究开发，品质评价和质量控制提供了科学依据。

High-performance liquid chromatographic coupled with variable wavelength detection （HPLC-VWD） has been developed for simultaneous determination of 5 analytes including ellagic acid, quercetin, kaempferol-3-O43-D-rutinoside, tiliroside and kaempferol, and high-performance liquid chromatographic with an evaporative light scattering detector （HPLC-ELSD） has been estab- lished to determine goshonoside-F5 in extract of Rubi Fructus. Chromatographic separations were carried out on an Agilent ZORBAX SB-CIs column （4. 6 mm ×250 mm, 5. 0 μm）. All calibration curves of reference standards revealed good linearity （R2 〉0. 999 5） within the concentration ranges tested. The method limits of detection ranged 0. 297-90. 144 ng and the method limits ofquantitation ranged 0. 990-300. 480 ng, respectively. Recoveries of 6 analytes were from 97. 11% to 101.7%, with RSD less than 2. 1%. The result shows that amounts of the 6 analytes in the samples from 16 localities were found to be different. The higher latitude of growing environment , the more ellagic acid in herb. The content of total flavonoids in sample from east localities were higher than that in middle and west localities, and the content of goshonoside-F5 in Bozhou, Anhui province was higher than others. This method was found to be simple, accurate, sensitive with good repeatability. Those results might serve as a sound foundation for further study, quality control and application of Rubi Fructus.