HPLC法测定云南花粉片中总黄酮醇苷的含量

Determination of Total Flavoniods in Yunnan Pollen Tablet by HPLC

目的：建立云南花粉片中总黄酮醇苷的HPLC测定方法。方法：以HPLC法定量分析总黄酮醇苷的含量。色谱条件：色谱LichrospherC-18（250mm×4．0mm，5μm），流动相：甲醇－0．4％磷酸溶液（50：50），检测波长：360nm^[1-2]，流速为1．0mL／min，进样量：10μL，以峰面积外标法定量。结果：槲皮素与山奈素的峰与其他色谱峰分离良好，槲皮素在3．10-18．60μg／ml范围内线性关系良好，回归方程为y=0．02670X-0．1489（r=0．9999），山奈素在6．15～36．90μg／ml范围内线性关系良好，回归方程为y=0．02406X-0．0942（r=0．9999）。槲皮素平均回收率为99．93％，RSD为1．40％，山奈素平均回收率为100．03，RSD为1．86％。结论：本方法结果准确，重现性好，可以作为云南花粉片中总黄酮醇苷含量的定量分析方法。

Objective:To establish a HPLC method for the determination of total flavonids in Yunnan pollen tablet.Methods: HPLC was used to make a quantitative analysis of total flavonoids.Chromatography lichrospher was C-18(250mm × 4.0,5μm).Mobile phase was methanol-0.4% phosphoric acid(50:50),with 360nm of detection wavelength,at a flow rate of 1.0mL/min.Sample volume was 10μL,with peak area quantified by external standard.Results: The peak of quercetion and kaempferol were well separated from other chromatographic peak.The quercetim showed a good linear at the range of 3.10~18.60μg/ml;regression equation was y=0.02670x-0.1489(r=0.9999).Kaempferol showed a good linear at the range of 6.15~36.90μg/ml;regression equation was y=0.02406x-0.0942(r=0.9999).The average recovery rate of the quercetin was 99.93%(RSD=1.40%).The average recovery rate of the kaempferol was 100.03%(RSD= 1.86%).Conclusion: This method is accurate and reliable with good reappearance and can be used for the quantitative analysis of the total flavonoids inYunnan pollen tablet.