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

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体积分数15%乙醇水溶液洗杂,最后用3 BV体积分数80%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0.69、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。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。

The optimal macroporous adsorption resin for enrichment and purification of gardenia yellow pigment(GYP) was determined by dynamic screening methods. The purification process conditions for GYP were optimized and amplified. The properties of GYP before and after purification and its inhibition effects on the low density lipoprotein(LDL) oxidation modification were compared. The results showed that NKA macroporous resin was the most suitable resin for the enrichment and purification of GYP. The optimal purification process conditions were that the mass concentration of sample was 0.72 g/L, flow rate of sample was 2 BV/h(BV is bed volume), sample volume was 27 BV, the solutions for washing impurities were 7 BV distilled water and 9 BV 15%(volumn fraction) ethanol, followed by desorption with 3 BV 80% ethanol at 2 BV/h. The color value of the purified GYP(GYP-2) was 5.14 times higher than that of without purification GYP(GYP-1). Compared with those of GYP-1, the absorbance ratio of geniposide and chlorogenic acid to 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. GYP-1 at different concentrations had a significantly inhibitory activity on the oxidation of LDL and the inhibitory effect presented a good dose-effect relationship. The inhibition effect of GYP-2 at the same concentration on the generation of conjugated dienes(CD) and malondialdehyde(MDA) was significantly stronger than that of GYP-1(P<0.01). In addition, the effects of GYP-2 on retarding spectral red shift, inhibiting changes of LDL appearance colors and microstructures during oxidation were also stronger than those of GYP-1.