摘要
目的建立保健食品中总黄酮醇苷的酸水解-高效液相色谱测定方法。方法采用甲醇回流提取总黄酮醇苷,酸水解法水解为槲皮素、山柰素和异鼠李素3种苷元,用高效液相色谱仪测定3种苷元含量,计算总黄酮醇苷含量。结果甲醇-25%盐酸(4∶1)在85℃水浴中回流提取0.5 h能使样品完全水解。最佳色谱条件:ODS C18色谱柱(150 mm×4.6 mm,5μm),流动相:甲醇-0.6%磷酸溶液(47∶53),柱温:35℃,检测波长:360 nm。槲皮素在4.766~95.52μg/m L、山柰素在5.052~101.0μg/m L、异鼠李素在2.027~48.66μg/m L的浓度范围内,线性关系良好(r=0.999 1、0.999 4、0.999 4);测定样品的相对标准偏差(RSD)为0.57%;平均加标回收率分别为98.25%、99.81%和101.8%,回收率试验的RSD分别为3.82%、3.31%和4.86%;槲皮素、山柰素检出限为0.50μg,异鼠李素为0.80μg,总黄酮醇苷为4.52μg。同一份样品采用本方法测定的总黄酮醇苷含量低于采用《保健食品检验与评价技术规范》(2003年版)中方法测定的总黄酮含量。结论本方法简便、结果稳定、重复性强,可应用于保健食品中总黄酮醇苷的含量测定。
Objective To develop a HPLC method for the determination of the total flavonol glycosides in health food by acid hydrolysis. Methods The sample was extracted by methanol and was hydrolyzed into 3 aglycones by acid hydrolysis.They were quercetin,kaempferide and isorhamnetin. Chromatographic separation was carried out by the method of HPLC.The content of total flavonol glycosides was calculated. Results Methanol-25% hydrochloric acid solution(4∶1) was added into selected samples, then 85 ℃ water bath heating and refluxing were conducted for 30 min. Optimum chromatographic conditions were as follows. Chromatographic column: ODS C18(150 mm ×4.6 mm,5μm). Mobile phase:used methanol-0.6% phosphoric acid(47∶53). Column temperature was set at 35 ℃. Detection wavelength was 360 nm. The linear ranges of quercetin,kaempferide and isorhamnetin were(4.766-95.52) μg/m L(r=0.999 1),(5.052-101.0) μg/m L(r=0.999 4) and(2.027-48.66) μg/m L(=0.999 r 4) respectively. The relative standard deviation of the samples was 0.57%. The recovery rate was 98.25%(RSD=3.82%),99.81%(RSD=3.31%) and 101.8%(RSD=4.86%). The detection limits of quercetin and kaempferide were both 0.50 μg,while Isorhamnetin was 0.80 μg,the total flavonol glycosides was 4.52 μg. The content of total flavonol glycosides detected by this method was lower than the content of the total glavonoids detected by the method provided by technical specification for inspection and evaluation of health food(2003 Edition). Conclusion This method proved to be simple,stable and had strong repeatability and could be used for the determination of total flavonol glycosides in health food.
出处
《预防医学》
2017年第6期574-578,共5页
CHINA PREVENTIVE MEDICINE JOURNAL
