反相高效液相色谱法测定亳菊提取物中木犀草素及芹菜素的含量

Determination of Luteolin and Apigenin in Chrysanthemum morifolium Extract by RP-HPLC

目的建立同时测定亳菊提取物中木犀草素及芹菜素的反相高效液相色谱检测方法。方法采用Restek PinnacleⅡC18（250mm×4.6mm,5μm）色谱柱,配制流动相为甲醇∶四氢呋喃∶0.3%H3PO4水溶液=42∶13∶56,在流速为1.0ml/min,检测波长为350nm,柱温为室温,进样量为10μl的情况下建立测定方法。结果木犀草素线性范围为3.99-99.75μg/ml（r=0.9996）,平均加样回收率为105.9%,RSD=1.5%;芹菜素线性范围为4.095-102.375μg/ml（r=0.9996）,平均加样回收率为103.7%,RSD=2.5%。结论用该方法测定亳菊提取物中木犀草素及芹菜素的含量,具有分析时间较短,分离度高的特点。

Objective To develop a reversed - phase high performance liquid chromatographic method for the simultaneous determination of luteolin and apigenin in Chrysanthemum morifolium extract. Methods The separation was carried out on a Restek Pinnacle Ⅱ C1s column（250 mm ×4.6 mm,5 μm）with MeOH： THF： 0.3% H3PO4 =42： 13： 56 as mobile phase at a flow rate of 1.0 ml/min. The absorbanee was monitored at 350 nm. The volume of injection was 10 μl. Results The linearity was good within the ranges of 3.99 μg/ml - 99.75 μg/ml （ r = 0. 999 6 ） for luteolin and 4. 095 μg/ml - 102. 375 μg/ml （ r = 0. 999 6 ） for apigenin. The average recoveries were 105.9% and 103.7% and the relative standard deviations （RSDs）were 1.5% and 2.5% respectively. Conclusion The method used for the determination of luteolin and apigenin in the Chrysanthemum morifolium extract has the characteristics of short retention and high resolution.