反相高效液相色谱法测定苦荞中的芦丁和槲皮素

Determination of Rutin and Quercetin in Tartary Buckwheat by Reversed-phase High Performance Liquid Chromatography

目的:建立一种苦荞黄酮中芦丁和槲皮素含量的测定方法。方法:分别以芦丁和槲皮素为标品,采用KromasilC18反相柱(4.6mm×300mm,5μm),分别测定水解和未水解苦荞黄酮样品中芦丁和槲皮素的含量。结果:在流动相为0.4%磷酸-甲醇(体积比为40∶60)、检验波长358nm、进样速度1.0mL/min、柱温20℃条件下,测定的芦丁含量的线性范围为0.025～0.125g/L(r=0.9993),槲皮素含量的线性范围为0.008～0.256g/L(r=0.9995)。槲皮素的加样回收率99.9%,变异系数为1.50%(x,n=5);芦丁的加样回收率98.5%,变异系数为1.16%(x,n=5)。水解样中槲皮素含量为(194.7±4.22)mg/g,未水解样中槲皮素含量为(8.96±0.26)mg/g,芦丁含量为(83.84±3.29)mg/g,变异系数(x,n=3)分别为:2.17%、2.90%和3.92%。结论:该方法简便、快速、准确,可用于苦荞黄酮中芦丁和槲皮素的含量测定。

Objective： To establish a method for the quantitative determination of rutin and quercetin in the flavonoids from tartary buckwheat. Methods： The contents of rutin and guercetin in hydrolysis and un-hydrolysis sample was detected with rutin and quercetin as the mark sample respectively. Results： By Kromasil C18-column with 0.4% of phosphoric acid-methanol （volume ratio of 40：60） as the mobile phase wave length 358 nm, the flow rate was 1.0 mL/min. tile column temperature was set at 20℃, the calibration curves were linear in the ranges of 0,025-0.125 g/L （r= 0.9993） and 0.008-0.256g/L （r=0.9995） for rutin and quercetin respectively. The average recoveries were 98.5% and 99.9%. The CV were 1.16% （x^-, n=5） and 1.50%（x^-, n=5） for rutin and quercetin respectively. The concents of quercetin were （194.7±4.22） mg/g （CV 2.17% ） in the hydrolysis sample. Quercetin were （8.96±0.26） mg/g （CV 2.90%）, rutin were （83.84±3.29） mg/g （CV 3.92%） in the un-hydrolysis sample （x^-, n=3） . Conclusion： This method is simple, rapid, accurate and reliable. It is useful for determination rutin and quercetin in the flavonoids from tartary buckwheat.