栀子黄色素富集纯化对其抑制LDL氧化修饰的影响

Effect of Gardenia Yellow Pigment Enrichment and Purification on its Inhibition of LDL Oxidation Modification

摘要:

通通过动态筛选方法确定栀子黄色素(Gardenia yellow pigment, GYP)最优吸附树脂，对其纯化工艺条件进行优化及放大测试，并对GYP纯化前后性质及其抑制低密度脂蛋白（Low density lipoprotein, LDL）氧化修饰进行了比较。结果表明，NKA大孔树脂最适合富集纯化GYP，最佳纯化条件为：上样液质量浓度0.72 g/L、流速2 BV/h（BV为床体积）、上样量27 BV，分别用7 BV去离子水、9 BV 15%乙醇水溶液洗杂，最后用3 BV80%乙醇以2 BV/h流速洗脱。在该条件下，纯化后的栀子黄色素（GYP-2）较纯化前栀子黄色素（GYP-1）色价提高了5.14倍，栀子苷、绿原酸与栀子黄色素吸光度的比值（OD1、OD2）分别从2.65、1.01降低到0.690、0.684。该工艺放大后效率稳定、效果良好。不同浓度GYP-1能抑制LDL氧化且呈现良好的量效关系。与GYP-1相比较，同等浓度GYP-2对共轭二烯（CD）、丙二醛（MDA）产生的抑制效果显著强于GYP-1 (P<0.01)，其减缓光谱红移、抑制LDL外观颜色及微观结构改变的效果也强于GYP-1。

Abstract:

The optimal macroporous adsorption resin for enrichment and purification of gardenia yellow pigment (GYP) was determined by dynamic screening methods. The purification conditions were optimized and amplified. The properties of before and after GYP purification and the inhibition effects of these two kinds of GYP on low density lipoprotein（LDL)oxidation modification were compared. The results showed as follows:NKA macroporous resin was the most suitable resin for the enrichment and purification of GYP. The optimal purification conditions were that sample mass concentration was 0.72g/L, sample flow rate was 2BV/h(BV isbed volume), sample volume was 27BV, the solutions for washing impurities were 7 BV distilled water and 9BV 15% ethanol , followed by desorption with 3BV 80% ethanol at a elution rate of 2BV/h. Compared withGYP before purification (GYP-1), the color value of GYP after purification(GYP-2) increased by 5.14 times, The absorbance ratio of geniposide, chlorogenic acid and GYP (OD1 and OD2) decreased from 2.65 and 1.01 to 0.69 and 0.684 under the above optimal purification conditions, respectively. The enlarged process of the optimal purification conditions had stable efficiency and good effect. The different concentrations of GYP-1 had a significantly inhibitory activity on oxidation of LDL and the inhibitory effectpresented a good dose-effect relationship. Compared with GYP-1, the inhibition efficiencies of GYP-2 with the same concentration were significantly enhanced against the generation of conjugated dienes (CD)and malondialdehyde (MDA) (P<0.01). The effects on retardingspectral red shift, inhibiting the changes of LDL appearance colors and microstructures during oxidation were also stronger than those of GYP-1.