RESUMO
To increase production efficiency of meat products, milk protein additives are often used. Despite a number of advantages, use of dairy ingredients involves a certain risk, namely the allergenic potential of milk proteins. A number of methods have been developed to detect milk-origin raw materials in foodstuffs, including immunological reference methods. This study presents newly developed immunohistochemical (IHC) methods for casein detection in meat products. Casein was successfully detected directly in meat products where sensitivity was determined at 1.21 and specificity at 0.28. The results obtained from the IHC were compared with the Enzyme-Linked Immuno Sorbent Assay (ELISA) and there was no statistically significant difference between the IHC and ELISA methods (p > 0.05). The correspondence between the methods was 72% in total. The highest correspondence was reached in frankfurters (90%), the lowest in canned pâté (44%).
RESUMO
The aim of the research was to verify the necessity of secondary fixation with osmium tetroxide in various types of meat products and evaluation of structural changes of products using different fixation procedures. The material for the study consisted of 11 types of meat products that were analyzed using a scanning electron microscope (SEM) with two different methods of chemical fixation. The first method included the usual processing of biological samples: glutaraldehyde primary fixation, the use of a buffer, secondary fixation by osmium tetroxide (OsO4), buffer, and dehydration using ethanol of increasing concentrations. The second method comprised the glutaraldehyde primary fixation and dehydration using the ethanol of increasing concentrations only. The results unambiguously suggest that the main difference between these methods is in fixation and visibility of fat. Our analysis principally suggests that fixation of the product with OsO4 allows the tracking of all components (fat droplets, muscle fibers, connective tissue) in meat products. At the same time, our results also support the possibility that the secondary fixation can be skipped during the analysis, where the main objection is an observation of lipid-free structures of the meat products (e.g., connection between muscle and starches or spices) or meat products with an insignificant amount of fat.
RESUMO
The aim of the study was to analytically evaluate quantum dots in immunohistofluorescence (IHF-QD) microscopic imaging as detectors of food allergens-peanut and wheat. The experiment was designed as two in silico experiments or simulations: (a) models of pastry samples were prepared with the addition of allergenic components (peanut and wheat protein components) and without the addition of allergenic components, and (b) positive and negative commercial samples underwent food allergen detection. The samples from both simulations were tested by the ELISA and IHF-QD microscopic methods. The primary antibodies (secondary antibodies to a rabbit Fc fragment with labeled CdSe/ZnS QD) were labelled at 525, 585, and 655 nm emissions. The use of quantum dots (QDs) has expanded to many science areas and they are also finding use in food allergen detection, as shown in the study. The study indicated that differences between the ELISA and IHF-QD microscopic methods were not observable among experimentally produced pastry samples with and without allergenic components, although differences were observed among commercial samples. The important value of the study is certainly the differences found in the application of different QD conjugates (525, 585, and 655). The highest contrast was found in the application of 585 QD conjugates that can serve for the possible quantification of present food allergens-peanuts and wheat. The study clearly emphasized that QD can be used for the qualitative detection of food allergens and can represent a reliable analytical method for food allergen detection in different food matrixes.
