Stereological analysis of pea protein in model samples
Keywords:Vegetable proteins, microscopy, immunohistochemistry, allergens, meat products
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.
Aguilera, J. M, Stanley, D. W. 1999. Microstructural principles of food processing & engineering. New York: Elsevier Science Publishers LTD, ISBN 978-0-8342-1256-5.
Baticz, O. 2001. Isolation and study of the functional properties of pea proteins. Nahrung/Food, vol. 45, no. 6, p. 399-401. DOI: https://doi.org/10.1002/1521-3803(20011001)45:6<399::AID-FOOD399>3.0.CO;2-0
Bedáňová, I., Večerek, V. 2007. Basics of statistics for studying veterinary medicine and pharmacy. 1st Ed. Brno: University of Veterinary and Pharmaceutical Sciences Brno, 130 p.
Flint, O. F., Meech, M. V. 1978. Quantitative determination of texturised soya protein by a stereological technique. The Analyst, vol. 103, no. 1224, p. 252-258. https://doi.org/10.1039/an9780300252 DOI: https://doi.org/10.1039/an9780300252
Glaser, J. R., Glaser, E. M. 2000. Stereology, morphometry, and mapping: the whole is greater than the sum of its parts. Journal of Chemical Neuroanatomy, vol. 20, no. 1, p. 115-126. https://doi.org/10.1016/S0891-0618(00)00073-9 DOI: https://doi.org/10.1016/S0891-0618(00)00073-9
Javůrková, Z. 2014. Food quality evaluation by image analysis. Brno, 133 p. Doctoral thesis on Faculty of Veterinary Hygiene and Ecology on University of Veterinary and Pharmaceutical Sciences Brno. Leader dissertation doc. MVDr. Bohuslava Tremlová, Ph.D.
Javůrková, Z., Pospiech, M., Zelenková, M., Kameník, J., Petrášová, M., Tremlová, B. 2015. Microscopic determination of bamboo fiber in meat products. Potravinarstvo, vol. 9, no. 1, p. 190-194. https://doi.org/10.5219/452 DOI: https://doi.org/10.5219/452
Kaláb, M., Wojtas, P. A., Miller, S. S. 1995. Microscopy and other imaging techniques in food structure analysis. Trends in Food Sciences and Technology, vol. 6, no. 6, p. 177-186. https://doi.org/10.1016/S0924-2244(00)89052-4 DOI: https://doi.org/10.1016/S0924-2244(00)89052-4
Lukáškova, Z. Řezáčová, Tremlová, B., Pospiech, M., Renčová, E., Randulová, Z., Steinhauser, L., Reichová, A., Bednář, J. 2011. Comparison of immunohistochemical, histochemical and immunochemical methods for the detection of wheat protein allergens in meat samples and cooked, dry, raw and fermented sausage samples, Food Additives and Contaminants Part A-Chemistry Analysis Control Exposure & Risk Assessment, vol. 28, no. 7, p. 817-825. https://doi.org/10.1080/19440049.2011.572292 DOI: https://doi.org/10.1080/19440049.2011.572292
Mellenthin, O., Galensa, R. 1999. Analysis of polyphenols using capillary zone electrophoresis and HPLC: detection of soy, lupin, and pea protein in meat products. Journal of agricultural and food chemistry. vol. 47, no. 2, p. 594-602. https://doi.org/10.1021/jf980749h DOI: https://doi.org/10.1021/jf980749h
Messiona, J. L., Assifaouia, A., Cayota, P., Saurela, R. 2012. Effect of pea proteins extraction and vicilin/legumin fractionation on the phase behavior in admixture with alginate. Food hydrocolloids. vol. 29, no. 2, p. 335-346. https://doi.org/10.1016/j.foodhyd.2012.03.003 DOI: https://doi.org/10.1016/j.foodhyd.2012.03.003
Modi, V. K., Mahendrakar, N. S., Narasimha Rao, D. Sachindra, N. M. 2004. Quality of buffalo meat burger containing legume flours as binders. Meat Science, vol. 66, no. 1, p. 143-149. https://doi.org/10.1016/S0309-1740(03)00078-0 DOI: https://doi.org/10.1016/S0309-1740(03)00078-0
Petrášová, M., Pospiech, M., Tremlová, B., Randulová, Z. 2015. Immunofluorescence detection of milk protein in meat products. Potravinarstvo, vol. 9, no. 1, p. 101-105, https://doi.org/10.5219/431 DOI: https://doi.org/10.5219/431
Pospiech M. 2008. Histochemical and immunohistochemical methods in food analysis with focus on soya protein detection. Brno, 133 p. Doctoral thesis on Faculty of Veterinary Hygiene and Ecology on University of Veterinary and Pharmaceutical Sciences Brno. Leader dissertation doc. MVDr. Bohuslava Tremlová, Ph.D.
Regulation (EU) No 1169/2011 of the European Parliament and of the Council of 25 October. Official Journal of the European Union. 2011, L 04, p. 18-63.
Řezáčová Lukášková, Z., Pospiech, M., Tremlová, B., Randulová, Z. 2011. Quantification of immunohistochemical detection of wheat protein in model samples. Proteins 2011, Zlín, p. 104-107.
Tremlová, B., Pospiech, M., Randulová, Z. 2013. Microscopic methods in the analysis of meat and meat products. 1st ed. Brno: knihovnicka.cz, 133 p.
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