免疫亲和柱净化-高效液相色谱法检测小麦中玉米赤霉烯酮  

HPLC determination of zearalenone in wheat with purification by immunoaffinity column

作　　者：王伟 刘安法 李明奇 邬冰 王彦斐 

机构地区：[1]河南中储粮质量检测中心有限公司,河南郑州450046 [2]北京中检维康技术有限公司,北京100044

出　　处：《粮食与饲料工业》2013年第4期59-60,65,共3页Cereal & Feed Industry

摘　　要：用50ml乙腈-水(体积比90∶10)萃取小麦试样,提取液稀释过滤后经含有玉米赤霉烯酮特异性抗体的Zearala Test免疫亲和柱层析净化,甲醇1.5ml洗脱,洗脱液用C18色谱柱(4.6mm×150mm,5μm)分离,流动相为乙腈-水(体积比60∶40)混合溶液,流速为1.0ml/min,采用荧光检测小麦中玉米赤霉烯酮。试验结果表明,玉米赤霉烯酮质量浓度在10.0～500.0μg/L范围内与色谱峰面积呈良好的线性关系,方法的检出限(3S/N)为5.0μg/L。应用此方法分析小麦加标样品,试样的平均回收率为83.7%～86.5%,测定值的相对标准偏差均小于5.0%,符合小麦中玉米赤霉烯酮的检测要求。Zearalenone was extracted from sample with acetonitrile and H20 （90 ： 10, v/v） mixture of 50 ml. The extract obtainedwas diluted and filtered. The filtrate was purified by passing through the Zearala Test immunoaffinity column containing zearalenone specificity antibody, and eluted with methanol of 1.5 ml, the elution was used for HPLC deternnnation. The C18 column （4.6 mm X 150 mm, 5μm）was used as separation column, and a mixture of acetonitrile and water （60 ： 40, v/v） was used as mobile phase at the flow rate of 1.0 mi/min. The zearalenone content in wheat was detected by fluorescence. The results showed that; the linear relationship between values of peak area and mass concentration was obtained in the range of 10.0-500.0μg/L. The detection limit（3S/N）found was 5.0 /μ/L. Application to the analysis of spiked wheat samples, the average recovery rate was 83. 7% - 86. 5%, and values of RSD was less than 5. 0%, which was coincidence with the requirements of zearalenone detection in wheat.

关 键 词：小麦 高效液相色谱法 免疫亲和层析净化 玉米赤霉烯酮 

分 类 号：TS210.7[轻工技术与工程—粮食、油脂及植物蛋白工程] O657.72[轻工技术与工程—食品科学与工程]