Pospiech M., Tremlová B., Renčová E., Randulová Z. A functional immunohistochemical method for soya proteins detection was developed. The procedure is based on the avidin-biotin complex (ABC) method that attains sufficient sensitivity. The method was verified by the analysis of the model samples of different forms of soya additives containing various concentrations of soya isolate. The detection limit of soya present in the model samples was 0.5%. Different possibilities of the background staining were tested. The best results were obtained with the background staining according to Calleja. The results were confirmed by the accredited indirect ELISA method. The method allows the identification of various forms of soya proteins such as isolates, texturates, concentrates, and flour.
Nowadays there are various vegetable protein additives intended for the manufacture of meat products in the food industry. These ingredients include both, plant-origin as well as animal-origin proteins. The most common vegetable additives include various types of flour, starch, fiber and plant protein. Among animal proteins, the most commonly used are plasma, collagen or milk protein. Milk protein is added to meat products due to its functional properties, such as emulsifying fats, improving the holding capacity of meat, improving juiciness, gel-forming capacity and affecting the taste of the product. Usage of these proteins, however, is currently limited by the effective legislation, not only in order to prevent consumer deception, but also because of their potential impact on consumers' health of. Thus, this issue has received considerable attention not only in the Czech Republic, but also globally. The main risk is the impossibility of selecting a suitable foodstuff for individuals with potential allergic reactions. The only option for allergic consumers to protect themselves is to strictly exclude the given allergen from their diet. Although the number of studies dealing with the reduction or loss of allergenicity is increasing, yet these practices are not common. Most of the population suffering from food allergies is thus still dependent on strict exclusion of foodstuffs causing adverse allergic reactions from their diet. Detection of allergens in foodstuffs is unfortunately quite difficult due to the fact that they occur in trace amounts and are often masked by different parts of the foodstuff. This research dealt with the detection of milk protein in meat products purchased in the market network of the Czech Republic, whereas declaration given by the manufacturer on the packaging for the small meat products purchased from the market was used to verify the detection of milk protein by the immunofluorescence method. 20 products were examined, these were selected with regard to the presence of milk protein that was declared by the manufacturer on the packaging. Method validation was performed by comparing the positive results from the investigated method with information on the packaging of the meat product. Milk protein was detected in 84.62 per cent of samples where the manufacturer declared the presence of milk or cheese on the package and additionally in 85.71 per cent of samples where the manufacturer declared the presence of milk protein. The results show that the immunofluorescence method is suitable for the detection of milk protein in meat products.
Randulová Z., Tremlová B., Řezáčová-Lukášková Z., Pospiech M., Straka I. (2011): Determination of soya protein in model meat products using image analysis. Czech J. Food Sci., 29: 318-321.The addition of plant proteins into meat products is nowadays a commonly used practice especially for the technological and economical reasons. Their properties have been known and used in meat products production for a long time. In the past, wheat protein or flour had been used most frequently, however, in these days they are being replaced by soya protein which has much more favourable properties in its use. Considering the possible misuse of raw materials of plant origin for the adulteration of meat products, the existence of highly sensitive and accurate procedures for their detection is needed including the determination of their content. Soya protein can be detected using various methods. In our work, an immunohistochemical method was used with image analysis for the quantification of soya protein. Model meat products with the addition of known amounts of soya protein in various forms were made for this experiment.
Pospiech M., Tremlová B., Renčová E., Randulová Z., Řezáčová Lukášková Z., Pokorná J. (2011) This work compares the commonly used immunochemical methods for soya protein detection and alternative microscopic methods. Immunochemical methods were represented by the competitive ELISA method. Histochemical and immunohistochemical methods were used for microscopical examination. From a group of 252 meat products, each sample was examined for soya proteins by ELISA, histochemical, and immunohistochemical methods. The products came from the following categories: cooked sausages, ham, dry cooked sausages, and fermented sausages. The results showed that the highest accuracy was achieved by immunohistochemical examination. However, in the category of cooked sausages, this result was not statistically significant. Since the results in the individual categories differed, our results demonstrate that one single method does not always provide reliable and completely objective results. Immunohistochemical methods seem to be the most suitable for the verification of the reference immunochemical method results and prevention of false results.
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