DM130大孔吸附树脂富集葎草中总黄酮的工艺研究

目的]研究DM130大孔吸附树脂纯化葎草总黄酮工艺并采用梯度洗脱HPLC法测定各洗脱组分中牡荆素、木犀草素和大波斯菊苷的含量。方法]通过单因素分析考察DM130大孔吸附树脂分离、纯化葎草总黄酮的最佳工艺条件。HPLC检测波长为350 nm,流速1 ml/min,温度25℃,色谱柱为Hanbon Kromasil C_8色谱柱（250.0 mm×4.6 mm,5.0μm）,流动相A为水相（甲醇：水：甲酸=30：70：0.1）,流动相B为有机相（0.1%甲酸甲醇,即每100 ml甲醇含0.1 ml甲酸）。结果]DM130树脂具体工艺条件为控制上样浓度0.2mg/ml,流速2.0 BV/h,上样液pH值为6,洗脱用水4.0 BV,50%乙醇共3.5 BV进行洗脱,洗脱流速2.0 BV/h;牡荆素进样浓度在0.16～20.00μg/ml,大波斯菊苷进样浓度在0.16～20.00μg/ml,木犀草素进样浓度在0.40～50.00μg/ml范围内与峰面积呈良好的线性关系。结论]DM130型树脂在此工艺条件下,纯化葎草总黄酮效果良好,总黄酮回收率可达近88%,各洗脱组分中牡荆素、木犀草素、大波斯菊苷含量不同。

Study on the Total Flavonoids of Humulus scandens(Lour.) Merr.Technology in DM130 Macroporous Resin Concentration

Objective] To study the technology for purification of flavonoids from Humulus scandens (Lour.)Merr. by DM130 macroporus adsorbtion resin and determine the content of cosmosiin,vitexin and luteolin by Gradient HPLC. Method] The proper technology parameters of purification of flavonoids were obtained through single factor test. Chromatographic separation was peformed in Hanbon Kromasil C_8 column(4.6 mm×250.0 mm,5.0 μm) at 25 ℃with an molibe phaseA consisting of CH_3OH-H_2O-CH_3COOH(30:70:0.1) and molibe phaseB of CH_3OH-CH_3COOH(1 000:1) at a flow rate of 1.0 ml/min, the molibe phase was monitored by a photodiode array detector at 350 nm. Result] The best technical parameter of DM130 macroporous resin were as follows, the content flavonoids was 0.2 mg/ml, adsorption flow rate was 2.0 BV/h, pH value was 6, after eluted with 4 BV distilled water ,the eluting regent 3.5 BV 50% ethanol and eluting rate 2.0 BV/h. The linear range of vitexin was 0.16-20.00 μg /ml, the linear range of cosmosiin was 0.16-20.00 μg /ml, and the linear range of luteolin was 0.16-20.00 μg /ml. Luteolin had a good linear relationship with peak area in the 0.40-50.00 ug/ml concentration range. Conclusion] The DM130 macroporus adorsption resin showed better comprehensive adsorption property, and the average recovery of the method was 88%, the contents of vitexin, luteolin and cosmosiin in each elution fraction group were different.