高效液相色谱法同时测定荷叶中6种黄酮类成分

Simultaneous Determination of Six Flavonoid Compounds in Lotus Leaves by High Performance Liquid Chromatography

为了探索同时测定荷叶茶及饮片中6种黄酮类成分（芦丁、金丝桃苷、紫云英苷、槲皮素、山奈素和异鼠李素）含量的方法,本研究以8种不同的荷叶样本为材料,采用高效液相色谱分析法对6种黄酮类成分进行了同时测定。样品经过前处理,以0.5%甲酸-水（A）和0.1%甲酸-乙腈（B）作为流动相进行梯度洗脱,流速为1mL/min,柱温为25℃,进样量为20μL,经Agilent TC-C18（2）（150mm×4.6mm,5μm）色谱柱分离,于360nm波长处检测,结果显示,芦丁、金丝桃苷、紫云英苷、槲皮素、山奈酚、异鼠李素6种黄酮类成分分别在1.6~160、1.8~180、2.16~216、1.4~140、2.12~212、1.6~160μg/mL浓度范围内有良好的线性关系（R2〉0.9992）,其检测稳定性、重复性、日内精密度、日间精密度以及加样回收率的RSD均小于2%。进一步用该方法对不同来源的8个荷叶样本进行检测,结果显示荷叶样本中6种黄酮类成分含量以槲皮素最高,并且以样本G荷叶茶（购自G公司,批号为130802）的槲皮素含量最高。本研究建立的同时测定6种荷叶黄酮类成分含量的方法快速、准确,可为荷叶有效成分的检测和质量控制提供技术支撑。

To simultaneously determine the contents of six flavonoids（rutin,hyperoside,astragalin,quercetin,kaempferol and isorhamnetin）in lotus leaves and related tea products,eight lotus leaf samples from different companies were detected by HPLC.The prepared samples were separated by Agilent TC-C18（2）（150 mm × 4.6 mm,5μm）at 25℃ with a step linear gradient using 0.5%formic acid-water（A）and 0.1%formic acid-acetonitrile（B）at a flow rate of 1.0 mL/min.The detection wavelength was 360 nm.Results showed that the concentration ranges of rutin,hyperoside,astragalin,quercetin,kaempferide and isorhamnetin were 1.6-160,1.8- 180,2.16-216,1.4-140,2.12-212 and 1.6-160μg/mL,respectively.Between these ranges,the linearity of the six flavonoids were well-presented（R 2 0.9992）in a certain concentration range.The RSD values for the stability,repeatability,intra-day and inter-day precisions and recovery rates were all less than 2%.Eight lotus leaf samples from different companies were detected by the above method.The content of quercetin was the highest among the six flavonoids,and was highest in Batch No.130802 from Company G.The method used in this study accurately detected the contents of six flavonoids in lotus leaves and tea products,and provided an effective way in which to determine the functional components and quality of lotus leaf products.