枳实提取物中类黄酮高效液相色谱法测定及其抗氧化作用探究.docVIP

枳实提取物中类黄酮高效液相色谱法测定及其抗氧化作用探究 　【摘要】 　　目的建立了枳实中柚皮苷、橙皮苷、新橙皮苷、芦丁、槲皮素含量的测定方法，同时测定了枳实提取物的抗氧化作用。方法采用HPLC法，色谱柱：Phenomenex Gemini C18柱(150 mm×4.6 mm，5 μm)，柱温37℃，流动相为乙腈-磷酸溶液，检测波长280，365 nm，流速1.0 ml/min 。采用DPPH·测定枳实提取物的自由基清除能力。结果进样量范围：柚皮苷为1.143～5.717 μg，橙皮苷为0.092～0.459 μg，新橙皮苷为1.179～5.895 μg，芦丁为0.024～0.118 μg，槲皮素为0.079～0.395 μg。柚皮苷、橙皮苷、新橙皮苷的平均回收率分别为99.604%，100.150%，100.249%。RSD分别为0.515%，4.390%，0.774%。枳实提取物对DPPH·清除作用是柚皮苷的17.6倍。结论HPLC测定法简便、准确，可用于测定枳实类黄酮含量，同时枳实提取物具有较强的自由基清除作用。 【关键词】 枳实； 类黄酮； 高效液相色谱法； 抗氧化作用 　　Abstract：ObjectiveTo establish an HPLC method for determination of naringin, hesperidin, neohesperidin, rutin and quercetin in Citrus aurantium L., and to study antioxidant activity .MethodsThe samples were extracted with ethanol. The chromatographic conditions were as follows: Phenomenex Gemini C18 column (150 mm×4.6 mm,5μm), column temperature at 37℃, mobile phase of acetonitrile-phosphate solution, detection wavelength at 280nm or 365nm, flow rate 1.0ml/min. In addition, DPPH radical-scavenging was employed to measure antioxidant activities. ResultsThe calibration curves were linear in the range of 1.143～5.717μg for naringin, 0.092～0.459μg for hesperidin, 1.179～5.895μg for neohesperidin, 0.024～0.118μg for rutin and 0.079～0.395μg for quercetin. The average recoveries of naringin, hesperidin, neohesperidin were 99.604%, 100.150% and 100.249% respectively. RSD were 0.515%, 4.390% and 0.774% respectively. The DPPH radical scavenging activity of C. aurantium L. flavoniods was 17.6 times of naringin. ConclusionThe method is simple, accurate for detecting flavoniods in C. aurantium L., and the C.aurantium L .extract has antioxidant activities. 　　Key words：Citrus aurantium L. ； Flavoniods ; High performance liquid chromatography; Antioxidant activity 　　枳实为芸香科植物酸橙Citrus aurantium L.及其栽培变种或甜橙Citrus sinensis Osbeck的干燥幼果，具有破气除痞、化痰消积的功效，是中医临床常用药物［1］。 　　关于测定枳实类黄酮的HPLC法大部分报道是测定柚皮苷、橙皮苷［2，3］，而对于新橙皮苷、芦丁、槲皮素的测定，少有报道，同时对于枳实类黄酮的抗氧化作用未见报道。本研究采用HPLC测定枳实中柚皮苷、橙皮苷、新橙皮苷、芦丁、槲皮素的含量，同时测定了枳实类黄酮对DPPH