HPLC法测定拟南芥中总黄酮含量的研究

为更好地开发和利用拟南芥中总黄酮及相关副产品,研究了检测拟南芥中总黄酮含量的方法。结果表明:以芦丁为标准品,建立了检测拟南芥中总黄酮含量的HPLC法。HPLC法色谱条件:C18色谱柱、35℃柱温、256nm检测波长、35%甲醇∶65%水(含0.4%磷酸)流动相、10μL进样量及10mL·min-1流速。标准品芦丁在该色谱条件下,峰型良好,在0.05~1.60 mg·mL-1时峰面积与质量浓度间呈良好线性关系,R2=0.999 7,平均加样回收率为99.90%,RSD小于3%(n=6),该方法适用于拟南芥中总黄酮的检测。

Determination of Total Flavonoids Content in Arabidopsis thaliana by High-Performance Liquid Chromatographic

In order to develop and use the flavonoids and related by-products in Arabidopsis thaliana, the deter- mine method was established for total flavonoids content. The results showed that taking rutin as the standard, high-performance liquid chromatographic method was established, mobile phase consisted on methanol （A）/ 0.4% orthophosphoric acid in distrilled water（B）and each solvent was filtered through a 0.45 μm pore size fil- ter and degassed in an ultrasonic bath before. The analysis of rutin was performed at 35℃ （column oven tem- perature）with 10 mL.min1 flow rate, the injection volume was 10 μL,and solvent A：B of rutin was 35%： 65% ,the chromatogram was detected at 256 nm. The peak area had a good linearity with concentration when R2 ： 0. 999 7, the recoveries of standard spiling was 99.90 % with RSD less than 3 %. So the method was suit- able for determination of total flavonoids in Arabidopsis thaliana.