高效液相色谱法同时测定保健食品中的安石榴甙和鞣花酸

Simultaneous determination of ellagic acid and punicalagins in health food by HPLC

目的建立同时测定保健食品中安石榴甙、鞣花酸的高效液相色谱法(HPLC)方法。方法采用迪马钻石Ⅱ(4.6 mm×250 mm,5μm)色谱柱,乙腈+0.2%磷酸水溶液为流动相梯度洗脱,在波长256 nm下进行检测。结果方法的线性范围:α安石榴甙24～240μg/ml;β安石榴甙56.0～560μg/ml;鞣花酸60.0～600μg/ml。相关系数r为0.999 7～0.999 9;检出限:α安石榴甙、β安石榴甙、鞣花酸分别为2.36、2.78、2.67μg/g;定量限:7.08、8.34、8.01μg/g。高低两个浓度水平加标回收率97.5%～104%;相对标准偏差(RSD)均<2.5%。结论本方法简便、快速、准确、重现性好,适用于保健食品中安石榴甙和鞣花酸的测定。

Objective To establish a method for the simultaneous determination of ellagic acid and punicalagins in health food by high performance liquid chromatography（HPLC）.Methods Chromatographic separation was carried out by using a Diamonsil C18（Ⅱ） column（4.6 mm×250 mm,5 μm）.The mobile phases were acetonitrile and 0.2% H3PO4 aqueous solution with an optimized gradient elution.The detective wavelength was 256 nm.Results The detection limits of α ellagic acid,β ellagic acid and punicalagins were 2.36,2.78 and 2.67 μg/g,respectively.The linear range for α ellagic acid,β ellagic acid and punicalagins were 24-240 μg/ml,56.0-560 μg/ml and 60.0-600 μg/ml,respectively,and all of which showed a correlation coefficient of over 0.999 7.The recoveries of three analytes at two spiked levels were ranged from 97.5%-104% and all of the relative standard deviation（RSD） were less than 2.5%.Conclusion This method was validated to be simple,rapid,accurate and reproducible for the determination of Ellagic Acid and punicalagins in health food.