| zeranolcomes from corn, peanuts, wheat, rice and corn gibberellic alcohol bacteria in non-standard storage when the secondary metabolites of corn gibberellic ketene reduction product, commodity name "livestock big strong" also known as "the right ring 14 ketone of phenol, the reduction product belongs to ray acid lactone class nonsteroidal assimilation hormone substances, have stronger to promote growth111.Effect of zeranol is a kind of better promote animal growth hormone-like substances, belong to the steroids of hormone-like chemicals in the kind of material, zeranol have a function to promote the synthesis of protein and protein efficiency so widely used in Europe and the United States and other developed countries, our country is a demonstration in China began in the late 20th century China. Later, after a lot of experiments and data that the zeranol in mammals have to accumulate in the body, and all have certain harm to both animals and humans.So explicitly in the developed countries it is forbidden to zeranol and hormone drugs for aquaculture , the ministry of agriculture in China no.193 announcements specified zeranol in harmful to human body, such as antibiotic drugs are banned for all food animals, as well as stipulated in the edible animal in the organization and may not be detected in the urine. And the current detection methods are mainly liquid, mass spectrometry and tandem mass spectrometry and other methods, but these methods are tedious, time is long, the operation personnel technical requirements is very high, so the method of enzyme-linked immune more easily by the public reception, because corn gibberellic alcohol enzyme-linked immune method is simple, practical stronger, better specificity, technical requirements of the operator is not high. More likely to be at the grass-roots level examination department to accept, feed companies, etc. Can also be used.1. Objective:to establish a rapid, effective, economical zeranol test kit for detecting the residues of ZER plant-based foods.2. Method:the indirect competitive ELISA method, this method is to adopt the principle of the specific antigen and antibody response, will need to detect smooth connection by using the method of biological substances and enzymes arose, and finally by enzyme and substrate produce different color and quantitatively detect the content of the material. Determination of the object can be antigen can be antibody also build corn gibberellic alcohol monoclonal antibody rapid detection of food of plant ZER content in the qualitative or half quantitative detection. The ELISA kit is the package is at the same time, fixed, dry and so on a series of debugging, making corn gibberellic alcohol kit reaction is more sensitive, more specific.3. Results:the experiment based on synthetic ZER package is original, ZER hapten for competition, both with a certain amount of resistance to corn gibberellic alcohol monoclonal antibody response, to establish ELISA method to detect ZER. When the package is concentration is 1:5000 is an ideal package is the concentration ratio, when the sheet resistance and enzymes two dilution ratio regulation to 1:500 and 2500, the kit of the most ideal color effect. However, when the sample solution with a single solution volume than 50:50 resistance enzyme-linked effect is the most ideal, the choice of substrate solution medium concentration of the substrate color, single resistance to determine the reaction time of 60 min, the optimal working concentration of 1:5000 antibody, enzyme mark 2, in which the optimal working concentration 1:5000. When the developed kit control the detection limit of 0.5 ng/mL, add recovery rate was between 60.0%～120.0%, the variation coefficient is less than 20%, under the condition of 2～8℃ can be saved for 3 months. |
