HPLC测定黄曲霉菌侵染后花生种子中6种芪化物  

Determination of six stilbenes in peanut seeds after invaded by Aspergillus flavus

作　　者：王琴飞[1] 应东山[1] 洪彩香[1] 李莉萍[1] 王明[1] 张如莲[1] 

机构地区：[1]中国热带农业科学院热带作物品种资源研究所/农业部华南作物基因资源与种质创制重点实验室,海南儋州571737

出　　处：《中国油料作物学报》2013年第4期428-435,共8页Chinese Journal of Oil Crop Sciences

基　　金：海南省自然科学基金(809034);中央级公益性科研院所基本科研业务费专项资金(中国热带农业科学院热带作物品种资源研究所)(1630032012025)

摘　　要：为建立一种同时检测受黄曲霉菌侵染后花生种子中白藜芦醇糖苷(Piceid)、白皮杉醇(Piceatannol)、白藜芦醇(Resveratrol)、两种白藜芦醇二聚体(ε-viniferin、δ-viniferin)和紫檀芪(Pterostilbene)的高效液相色谱(HPLC)方法,探索不同种植年度和地区的花生种子,在受黄曲霉菌侵染后6种芪化物的合成规律,采用PurospherSTAR RP-18e色谱柱(Hiber150×4.6mm,5μm);流动相为乙腈、水和2%甲酸水溶液三元梯度洗脱,柱温为30℃,检测波长306nm,流速1mL/min。结果显示,6种芪化物线性范围在1～100mg/L之间与峰面积的线性关系良好,相关系数在0.999 0～0.999 9之间;精密度、重复性和稳定性的标准偏差(RSD)都低于4.7%;检出限为0.01～0.05mg/L;加标回收率在95.2%～102.0%之间(n=3);所测两个品种的花生种子在受到黄曲霉菌侵染后,6种芪化物总含量在不同种植年度间变化差异较小,分别为1.23和0.40μg/g;在不同种植地区间变化差异较大,分别为33.69和71.08μg/g,品种间芪化物合成量与侵染率呈负相关。To understand the synthesis of stilbene in different years and regions of peanuts after Aspergillus flavus infection,simple HPLC method was developed on simultaneous determination of six stilbenes(resveratrol,piceid,piceatannol,ε-viniferin,δ-viniferin,and pterostilbene) in peanut.HPLC separation was achieved on a Purospher STAR RP-18e column(Hiber150×4.6mm,5μm) at 306 nm,30℃.A mobile phase of acetonitrile,water and 2% formic acid aqueous solution at a gradient elution with flow rate of 1mL/min was used.The method exhibited good linear relationships between the peak area and the concentrations range of 1 to 100 mg/L for the six stilbenes.The correlation coefficients were between 0.999 0 to 0.999 9.RSD of this method was less than 4.7%,the detection limits were between 0.01 and 0.05mg/L.The average recoveries were 95.2% to 102.0%.Four peanut varieties from different producing area and planting year were used to validate this method.Results indicated that the total amount of stilbenes varied slightly within the same producing area,with a value of 1.23 and 0.4μg/g respectively.However,significant differences were observed for the same varieties from different areas,possessing a value of 33.69 and 71.08μg/g respectively.A negative correlation relationship was also observed between the total amount of stilbenes and infection rate.The method is suitable for rapid and accurate determination of resveratrol,piceid,piceatannol,ε-viniferin,δ-viniferin,and pterostilbene from peanut seeds infected by Aspergillus flavus,and could be used for development of functional food and resistant breeding or varieties screening study of peanuts.

关 键 词：花生 白藜芦醇 芪化物 高效液相色谱法(HPLC) 

分 类 号：R151.3[医药卫生—营养与食品卫生学]