Development of Enzyme-Linked Immunoassay Systems for Monitoring the Content of the Main Non-Meat Additives in Meat Products

2021 ◽  
Vol 37 (5) ◽  
pp. 117-122
Author(s):  
E.A. Zvereva ◽  
O.D. Hendrickson ◽  
B.B. Dzantiev ◽  
A.V. Zherdev
Keyword(s):  
Trypsin Inhibitor ◽  
Room Temperature ◽  
Meat Products ◽  
Reaction Medium ◽  
Soybean Trypsin Inhibitor ◽  
Russian Science ◽  
Additional Advantage ◽  
Relative Standard ◽  
Total Analysis ◽  
Kinetic Mode

Abstract-Methods for control of the content of non-meat components (connective tissue of animals, eggs, soybeans) in meat products have been developed based on competitive enzyme immunoassay of biomarker proteins: collagen, ovalbumin, and soybean trypsin inhibitor. Polyclonal rabbit antibodies against the biomarkers were produced. Screening of the following analysis conditions was carried out using the most affine preparations: the duration of the stages, the concentrations of the reagents, the composition of the reaction medium to ensure the completeness and the minimum limits of detection of analytes. It was shown that the immunochemical stage can be transferred to the kinetic mode and reduced to 20 min, while the total analysis took only 1 h. The selected conditions synchronized detection stages and provided the same signal amplitudes from all three biomarkers. An additional advantage of the method is that the analysis can be carried out at room temperature. The detection limits for collagen, ovalbumin and soybean trypsin inhibitor in the final extracts were 0.025 μg/mL, 0.012 μg/mL, and 0.001 μg/mL, respectively. Approbation of the method showed the reliability of the conclusions about the composition of the tested meat products; the relative standard deviation in the analysis of the monitored biomarkers was 8-10%. Key words: enzyme-linked immunoassay, meat products, collagen, ovalbumin, soybean trypsin inhibitor The work was financially supported by the Russian Science Foundation (project no. 19-16-00108).

DEVELOPMENT OF IMMUNOENZYME ANALYTICAL SYSTEMS FOR CONTROL OF THE CONTENT OF MAIN NON-MEAT COMPOUNDS IN PROCESSED MEAT PRODUCTS

2021 ◽  
Vol 1 (19) ◽  
pp. 322-324
Author(s):  
E.A. Zvereva ◽  
A.V. Zherdev ◽  
B.B. Dzantiev
Keyword(s):  
Connective Tissue ◽  
Enzyme Immunoassay ◽  
Trypsin Inhibitor ◽  
Meat Products ◽  
Soybean Trypsin Inhibitor ◽  
Processed Meat

Methods have been developed to control the content of non-meat components (connective tissue of animals, eggs, soy) in processed meat products, based on enzyme immunoassay of biomarker proteins – collagen, ovalbumin, soybean trypsin inhibitor.

scholarly journals Immunoreactivity of Lupine and Soybean Allergens in Foods as Affected by Thermal Processing

Foods ◽  
2020 ◽  
Vol 9 (3) ◽  
pp. 254 ◽  
Author(s):  
Caterina Villa ◽  
Mónica B. M. V. Moura ◽  
Joana Costa ◽  
Isabel Mafra
Keyword(s):  
Trypsin Inhibitor ◽  
Thermal Processing ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Meat Products ◽  
Soybean Trypsin Inhibitor ◽  
Thermal Treatments ◽  
Food Matrix ◽  
Soybean Proteins ◽  
Sds Page

Lupine and soybean are important technological aids for the food industry. However, they are also capable of inducing severe allergic reactions in food-sensitized/allergic individuals. In this context, this work intended to study the combined effects of thermal processing and food matrix on the immunoreactivity of lupine and soybean proteins used as ingredients in bakery and meat products, respectively. For this purpose, the effects of baking, mild oven cooking, and autoclaving on the protein profiles were evaluated, using model mixtures simulating the production of lupine-containing breads and soybean-containing cooked hams/sausages, by native- and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and immunoblotting using specific antibodies. The results showed that lupine gamma-conglutin immunoreactivity was slightly decreased in wheat flour mixtures compared to rice, but it was more pronounced in baked products. In meat mixtures, substantial protein fragmentation was noted after autoclaving, with decreased immunoreactivity of soybean trypsin inhibitor. The analysis of 22 commercial products enabled the identification of lupine gamma-conglutin in four bakery samples and soybean trypsin-inhibitor in five sausages, and further differentiated autoclaved from other milder thermally treated products. Generally, the immunoreactivity of target proteins was reduced by all the tested thermal treatments, though at a higher extent after autoclaving, being slightly altered by the food matrix.

Factor X Activation by washed human platelets

1979 ◽  
Author(s):  
E van Wijk ◽  
L Kahlé ◽  
J ten Cate
Keyword(s):  
Human Factors ◽  
Trypsin Inhibitor ◽  
Calcium Ions ◽  
Factor Xa ◽  
Human Platelets ◽  
Soybean Trypsin Inhibitor ◽  
Thrombin Inhibitor ◽  
Chromogenic Substrate ◽  
Factor X ◽  
Factor X Activation

In a system of washed human platelets, Ca2+and purified human factors X anc II, a sufficient amount of thrombin is generated in about 10 minutes to aggregate the platelets. This thrombin is formed through the activation of FX by the platelets. In a system with either FX or FII present, no aggregation occurs. In addition no aggregation is observed when hirudin, a specific thrombin inhibitor, or when soybean trypsin inhibitor, which inhibits factor Xa, are added to the mixture. The formation of factor Xa can be monitored indirectly through the generation of thrombin, in the presence of an excess of prothrombin, using a thrombin sensitive chromogenic substrate. When washed platelets are incubated with FX alone for 10 minutes, no aggregation occurs and after the addition of prothrombin aggregation starts within 6 minutes. These findings confirm that washed platelets possess a factor X activating property. The generation of FXa proceeds in the absence of added Ca2+, whereas in the presence of Ca2+factor Xa activity reaches a maximum in 3 minutes, whereafter the activity progressively decreases. This may be due to the binding of Xa to the platelets in the presence of calcium ions.

ICP-MS AND ICP-AES ANALYSIS OF PLANT REFERENCE MATERIALS

2019 ◽  
Vol 85 (6) ◽  
pp. 11-24
Author(s):  
I. V. Nikolaeva ◽  
A. A. Kravchenko ◽  
S. V. Palessky ◽  
S. V. Nechepurenko ◽  
D. V. Semenova
Keyword(s):  
Reference Material ◽  
Mass Spectrometer ◽  
Reference Materials ◽  
Atomic Emission ◽  
Icp Ms ◽  
Lithium Metaborate ◽  
Relative Standard ◽  
Microwave System ◽  
Total Analysis ◽  
Siberian Pine

Two methods — ICP-MS and ICP-AES are used for certification of the new reference material — needles of Siberian pine (NSP-1). Techniques of the analysis include decomposition of plant samples in two different ways: acid digestion in a microwave system MARS-5 and lithium metaborate fusion followed by ICP-MS and ICP-AES analysis of the solutions. Simultaneous determinations of all the elements were carried out in low, medium and high resolution using SF-mass-spectrometer ELEMENT and atomic-emission spectrometer IRIS Advantage with external calibrations and internal standards (In — ICP-MS, Sc —ICP-AES). Middle and high resolutions of ICP mass spectrometer were used for interference corrections. Data obtained by ICP-MS and ICP-AES with different decomposition techniques are in good agreement. The ICP-MS and ICP-AES techniques have been validated by the analysis of three plant reference materials: LB-1 (leaf of a birch), Tr-1 (grass mixture) and EK-1 (Canadian pondweed). These techniques were used for the determination of 38 elements in the new reference material NSP-1. Relative standard deviations for most of the determined elements were below 10%. Combination of ICP-MS and ICP-AES techniques for certification of the new reference material makes it possible to expand the set of elements to be determined and to reduce the total analysis time.

scholarly journals Preparation of Li3PS4–Li3PO4 Solid Electrolytes by Liquid-Phase Shaking for All-Solid-State Batteries

2021 ◽  
Vol 2 (1) ◽  
pp. 39-48
Author(s):  
Nguyen H. H. Phuc ◽  
Takaki Maeda ◽  
Tokoharu Yamamoto ◽  
Hiroyuki Muto ◽  
Atsunori Matsuda
Keyword(s):  
Solid Solution ◽  
Solid State ◽  
Liquid Phase ◽  
Room Temperature ◽  
Reaction Medium ◽  
Magic Angle Spinning ◽  
Resonance Spectroscopy ◽  
Synthesis Methods ◽  
Magic Angle ◽  
Angle Spinning

A solid solution of a 100Li3PS4·xLi3PO4 solid electrolyte was easily prepared by liquid-phase synthesis. Instead of the conventional solid-state synthesis methods, ethyl propionate was used as the reaction medium. The initial stage of the reaction among Li2S, P2S5 and Li3PO4 was proved by ultraviolet-visible spectroscopy. The powder X-ray diffraction (XRD) results showed that the solid solution was formed up to x = 6. At x = 20, XRD peaks of Li3PO4 were detected in the prepared sample after heat treatment at 170 °C. However, the samples obtained at room temperature showed no evidence of Li3PO4 remaining for x = 20. Solid phosphorus-31 magic angle spinning nuclear magnetic resonance spectroscopy results proved the formation of a POS33− unit in the sample with x = 6. Improvements of ionic conductivity at room temperature and activation energy were obtained with the formation of the solid solution. The sample with x = 6 exhibited a better stability against Li metal than that with x = 0. The all-solid-state half-cell employing the sample with x = 6 at the positive electrode exhibited a better charge–discharge capacity than that employing the sample with x = 0.

scholarly journals PSV-25 Detection of heart and aorta tissue peptide markers by the multiple-reaction monitoring method

2020 ◽  
Vol 98 (Supplement_4) ◽  
pp. 362-363
Author(s):  
Daniil Khvostov ◽  
Natalya Vostrikova ◽  
Irina M Chernukha
Keyword(s):  
Amino Acid ◽  
High Performance ◽  
Multiple Reaction Monitoring ◽  
Meat Product ◽  
Meat Products ◽  
Amino Acid Sequences ◽  
Composition Analysis ◽  
Reaction Monitoring ◽  
Russian Science ◽  
Monitoring Method

Abstract Functional, particularly personalized meat-based foods are of more in demand by a consumer today. Functional additives, such as plant components and animal proteins from bovine or porcine tissues have been successfully used. With many ingredients added to foods, it is important to provide quality and composition monitoring to confirm the products’ authenticity, to identify undeclared or rarely used types of raw meat in product formulations. For example, if animal heart tissue is a component of a product formulation or if aorta tissue presents in a product due to improper trimming. Different methods are used to identify raw materials, including new approaches in proteomics and peptidomics that are considered the most effective modern methods nowadays. The purpose of the study is meat product composition analysis and special biomarker peptide identification to confirm the presence of heart and aorta tissue in a finished meat product. Over 20 amino acid sequences were checked based on earlier obtained data. Those amino acid sequences were analyzed with a high-performance liquid chromatography with mass spectrometric detection as described. The MS settings were selected using the Skyline. Signal-to-Noise ratio (S/N) over 10 units were used to choose the best peptide candidates. Seven peptides were found in porcine hearts. The best candidate was peptide VNVDEVGGEALGR (S/N - 73.10±5.3) from β-Hemoglobin. Two marker peptides from serum albumin were selected for pork aorta: TVLGNFAAFVQK (S/N 53.51±2.4) and EVTEFAK (S/N 31.69±4.1). These biomarkers showed the best detection and specificity. The multiply reaction monitoring method made it possible to identify the most/best specific peptides—biomarkers that could confirm the heart and/or aorta in meat products. The method can be used for comparative research or identification of best peptides that are specific to any type of animal tissue. The work was supported by the Russian Science Foundation, project no. 16-16- 10073.

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