Marcin A, Fencík R, Belicková E, Siklenka P
Institute of Experimental Veterinary Medicine, Kosice, Slovak Republic.
Vet Med (Praha). 1995 Jul;40(7):227-31.
A simple immunological method is described by which it is possible to detect the wheat flour according to wheat protein in the heat-processed meat products. The wheat proteins were extracted with a buffer TRIS-NaCl (0.02 mol/l, 0.5 mol/l) from: a) wheat flour; b) model meat products with addition of wheat flour (3.2, 3.2, 3.6, 0.8, 0.4 and 0.2%); c) commercial meat products containing wheat flour (sausages-S, P, PF). The aliquots of the extracts were subjected to an immunological dot-blot test. The extracts from the wheat flour and the model meat products were used for calibration of this method. The quantitative evaluation of the test was based on the spectrophotometric determination of the quantity of the violet stain on the nitrocellulose (NC) strip surfaces. The strips were immersed into the acetone and the absorbance were determined at 580 nm. The screening of the samples by dot-blot procedure proved to be an efficient way of the wheat flour determination in the cooked meat products at 0.2% level (i.e. 0.375 ng of wheat protein applied to the NC strip). Since the wheat antibody (primary antiserum) has no crossreactivity with pork or beef, this technique can be used for the detection of plant proteins in the mentioned mixtures.
本文描述了一种简单的免疫学方法，通过该方法可以根据热处理肉制品中的小麦蛋白来检测小麦粉。用TRIS-NaCl缓冲液（0.02 mol/l，0.5 mol/l）从以下物质中提取小麦蛋白：a）小麦粉；b）添加了小麦粉（3.2%、3.2%、3.6%、0.8%、0.4%和0.2%）的模型肉制品；c）含有小麦粉的市售肉制品（香肠-S、P、PF）。将提取物的等分试样进行免疫斑点印迹试验。用小麦粉和模型肉制品的提取物对该方法进行校准。试验的定量评估基于对硝酸纤维素（NC）条带表面紫色污渍量的分光光度测定。将条带浸入丙酮中，并在580 nm处测定吸光度。通过斑点印迹法对样品进行筛选被证明是一种在熟肉制品中检测小麦粉的有效方法，检测水平为0.2%（即应用于NC条带的0.375 ng小麦蛋白）。由于小麦抗体（一级抗血清）与猪肉或牛肉无交叉反应，该技术可用于检测上述混合物中的植物蛋白。
