Abstract
With the growing popularity of various plant proteins used as raw materials for meat production, interest of manufacturers to extend the range of such raw materials is increasing as well. Manufacturers are trying to minimize the cost of manufacturing their products with simultaneous preserving the nutritional value of their products to the maximum extent possible. Such cheaper raw materials, which are also nutritionally rich, include pea protein. Another advantage for manufacturers is the fact that legislation does not order them to indicate pea protein presence in case of its addition, as it does for other allergenic ingredients, although this legume contains storage proteins which can cause a variety of allergic reactions, just like other legumes. Currently no method used for its qualitative determination has been described in literature, let alone its quantitative determination. Our work describes a possible method that can be applied for its quantification. It is a stereological method applied to microscopic sections stained by immunohistochemical staining based on the avidin-biotin complex using monoclonal legumin (1H9) as the primary antibody. The stereological method is based on geometry, it applies knowledge of geometry to analyze a sample of diverse origin, size and internal structure. Despite potential shortcomings in staining microscopic preparations, stereology allows us to perform quantification based on knowledge of morphology of the observed structures. This work describes a procedure of a known pea protein addition quantification in model meat products by means of Ellipse software. Pea protein quantification was performed in two ways. In the first case ten microimages of all sections prepared were examined, while in the second case one scan of the entire section was analyzed. Based on the results, Spearman's correlation coefficient was calculated, which confirmed our assumption of correlation between the protein added into the product and the measured area in microimages. In both ways Spearman's correlation coefficient was rSp = 1000. We obtained regression equations in MS Excel, which can be used for calculation of pea protein addition based on measured area of this protein in microscopic section.
Highlights
In the meat industry, raw materials in the form of vegetable proteins, which are used as a meat substitute, are very common (Modi et al, 2004)
This motivates the producers’ efforts to replace them with other vegetable proteins that are not ranked among the allergens that must be indicated in harmony with the aforementioned legislation
The results indicate that with increasing addition of the proteins increases the measured area in section by lowest concentration (0.1 percentage)
Summary
Raw materials in the form of vegetable proteins, which are used as a meat substitute, are very common (Modi et al, 2004). During meat production is frequently used various vegetable and animal proteins. Meat product contain from animal - origin proteins as plasma, collagen or milk protein (Petrášová, 2015). Some of these vegetable proteins are classified as allergens in the legislation (Regulation (EU) No 1169/2011). Pea protein belongs among the most common ones (Baticz, 2001). Pea proteins include storage proteins which can cause allergic reactions. The literature identifies analysis of polyphenols characteristic of certain legumes and HPLC method which can detect up to 0.1% addition of soy protein in a meat product, as potential detection methods. Other possible methods that can be used to detect vegetable protein are microscopic methods
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