【作者】 王蕾；

【导师】 高腾云；

【作者基本信息】 河南农业大学 ， 兽医， 2008， 硕士

【摘要】 黄曲霉毒素是一种来源于自然界中霉菌的毒素,已经被证明为导致人类癌症的一类致癌源。黄曲霉毒素M1则是黄曲霉毒素B1的代谢产物,存在于食用了黄曲霉毒素B1污染饲料的动物的乳中,各国都对乳中黄曲霉毒素M1进行了严格限量。关于黄曲霉毒素B1的预防、去除和解毒的研究工作,显得非常重要。虽然国内外对黄曲霉毒素的去除和解毒进行了大量的研究,可迄今为止还没有一种可供工业上大规模应用的理想的解毒方法。所以,及时对食品中黄曲霉毒素进行定量检测,是目前最大限度地降低黄曲霉毒素危害的最好措施。本研究拟建立起免疫亲和柱—荧光光度法快速测定牛奶中黄曲霉毒素M1的方法。试样经过离心、脱脂、过滤后,滤液经过键合有黄曲霉毒素M1特殊抗体的免疫亲和柱净化,此抗体对黄曲霉毒素M1具有专一的识别能力,黄曲霉毒素M1键合在分离柱中的抗体上。用甲醇与水为10:90的混合液将免疫亲和柱上杂质除去;以甲醇与水之比为80:20的混合液通过分离柱洗脱,加入溴溶液衍生,以提高测定灵敏度。衍生化后的洗脱液于荧光光度计中测定黄曲霉毒素M1。通过选择最佳激发波长和发射波长;对阴性牛奶样品添加特定量的标准样品,进行平行试验,判断该方法的精确度;进行三个水平的标准样品回收率试验;确定检出限。从而评判方法的准确性和精确性。采用免疫亲和柱——荧光光度法检测牛奶中的黄曲霉毒素M1,检测低限为0.01μg/kg,在0.1～1.0μg/kg范围内,回收率为89.6%～96.1%,变异系数为0.52%～5.8%,分析一个样品的时间小于30min,分析过程中不使用黄曲霉毒素M1标准物质。本方法具有准确、简单、快速、安全等特点,可以满足批量样品的检测需要。被检测的青贮料、粗饲料、全混合日粮样品中黄曲霉毒素B1平均含量分别为8.28μg/kg、3.11μg/kg和6.56μg/kg,属于轻度污染。50份奶样中,阳性最高值为0.203μg/kg,阳性最低值为0.003μg/kg,阳性平均值为0.089μg/kg,均符合我国目前的限量标准。奶牛样品中黄曲霉毒素M1与饲料中黄曲霉毒素的污染有关,奶牛通过饲料途径食入的黄曲霉毒素B1,约有1.35%以M1的形式进入乳中。更多还原

【Abstract】 Aflatoxin has been proved a reason resulting in cancer in human being. Aflatoxin M1 is the metabolize production of aflatoxin B1.Aflatoxin M1 in milk comes from animals fed feeds have been polluted by aflatoxin B1.So the effort to prevent and dispose aflatoxin B1 is very important.Although many studies have been carried out inside and outside,so far there isn’t an ideal means applied in industry.At present the best measure to reduce the harm of aflatoxin is to detect aflatoxin in food in time.Recently,high sensitive instrument analysis means and rapid effective immunity means have been applied in detection.Aflatoxin M1 in milk are extracted and separated by immunoaffinity column.Then the column in washed with methanol water(10:90) twice and the concentrated aflatoxin M1 are eluted with methanol water(80:20).The aflatoxin M1 is determined by fluorometer with bromine.Test portions and spiked at 0.1,0.5,1.0μg/kg, recoveries are 89.6%～96.1%,variable coefficient are 0.52%～5.8%,the lowest.It is 30 minutes for analysis one sample.The average content of aflatoxin Blin silage,coarse fodder and blend diet is 8.28μg/kg, 3.11μg/kg,and 6.56μg/kg.respectively,the pollution is light.50 samples have been detected are all accorded with the limited standard in our country,the maximum of positive is 0.203μg/kg,the minimum is 0.003μg/kg,and the average value is 0.089μg/kg.There is relationship between aflatoxin M1 in milk and aflatoxin in feed.There is about 1.3% aflatoxin in milk comes from feed.更多还原