Analytical methods for gelatin differentiation from bovine and porcine origins and food products.

Usage of gelatin in food products has been widely debated for several years, which is about the source of gelatin that has been used, religion, and health. As an impact, various analytical methods have been introduced and developed to differentiate gelatin whether it is made from porcine or bovine sources. The analytical methods comprise a diverse range of equipment and techniques including spectroscopy, chemical precipitation, chromatography, and immunochemical. Each technique can differentiate gelatins for certain extent with advantages and limitations. This review is focused on overview of the analytical methods available for differentiation of bovine and porcine gelatin and gelatin in food products so that new method development can be established

Identification polypeptide biomarkers of porcine skin gelatin by two-dimensional electrophoresis

The peptide composition of gelatin is known to vary very common that the electrophoretic pattern of gelatin from one source differs from another source even for the same raw material. Therefore, the present study aimed to use proteomics field to identify gelatin polypeptides biomarker for depending on the condition under which collagen is hydrolyzed. Hence, it is porcine skins. The polypeptides obtained for porcine skin gelatins can be used as reference in future to detect the origins of gelatin added in the processed food. We compared porcine skin gelatin samples obtained from three producers. Total average numbers of polypeptides of porcine skin gelatins from company A, B and C were 303 ± 2.8, 285.5 ± 3.5 and 270.5 ± 4.9 spots respectively. 10 biomarkers were identified and presented in all different companies. We also did a mixture of porcine and bovine skin gelatin to detect the presence of these 10 biomarkers. The level of adulteration that could be detected was as low as 1.0% w/w

Chemical and functional properties of bovine and porcine skin gelatin

The ability to compare bovine and porcine skin gelatin based on their amino acid composition, polypeptides pattern, bloom strength, turbidity and foaming properties were investigated. Amino acid composition of both gelatin showed that the content of glycine, proline and arginine in porcine gelatin were higher than bovine gelatin. However, the polypeptides pattern between both gelatin is closely similar. The bloom strength of porcine gelatin was higher than bovine gelatin from pH 3 to pH 10. Both gelatin possessed highest bloom strength at pH 9. The lowest bloom strength of bovine gelatin was at pH 3 while porcine gelatin at pH 5. The highest turbidity of bovine gelatin obtained at pH 7 while porcine gelatin at pH 9. Foam expansion and foam stability of bovine gelatin were higher than porcine gelatin at all concentrations

Monoclonal antibody-based enzyme immunoassay for detection of porcine plasma in fish surimi

Detection of porcine plasma using indirect ELISA was developed using mAb B4E1 for the prevention of their usage in human food that creates religious and health conflicts. The immunoassay has a CV<20% and did not cross-react to other meat and non-meat proteins. The sensitivity of the assay is 0.25% (w/w) of porcine plasma in spiked raw and cooked fish surimi. The assay did not produce a false positive result for any of the commercial fish surimi tested that were not contain porcine plasma. Determination of a 60-kDa antigenic protein of porcine blood using Western Blot confirmed its presence in the plasma fraction of the porcine blood. Further proteomic analysis involving liquid chromatography-tandem mass spectrometry (LC-MS/MS) revealed the 60-kDa protein to be porcine serum albumin

Authentication approach using enzyme-linked immunosorbent assay for detection of porcine substances

Food manufacturers across the world commonly add animal substances in their food products. Some food products may contain porcine substances including pork, gelatine, blood and pepsin. These substances significantly affect the texture, colour or taste of the end products. Aside from enhancing sensory qualities, additional ingredients also contribute to preservation, bulk and nutrition. However, the inclusion of porcine substances might not be suitable among certain communities. One primary concern is fraud labelling which includes hiding the addition of porcine substances in food. Therefore, analytical techniques such as enzyme-linked immunosorbent assay (ELISA) have been developed to detect the porcine proteins in food. The ELISA delivers specificity and sensitivity in detecting the targeted animal species in food. This review provides an overview of the ELISA technique which has been developed for potential detection of porcine substances in laboratory-prepared food samples and commercial food products

Development of monoclonal antibodies against porcine blood for detection in fish-based products

Animal blood mainly from porcine and bovine have been used in human food as a binder, gelling agent, emulsifier, coloring agent and iron’s supplementary in most meat and fish-based products. However, certain communities including Muslims and Jews are strictly prohibited to consume animal blood in food due to prohibition assigned by each religious and related to health and personal matter. The addition of porcine plasma in fish surimi has been highlighted by the local authority in Malaysia. To date, a specific method to detect porcine plasma in food is unavailable. In this study, we have developed monoclonal antibodies (mAbs) against heated soluble proteins (HSPs) of porcine blood using fusion technology for detection of porcine plasma in fish surimi. Specificity of mAbs against blood, non-blood (meat and nonmeat) and commercial animal plasma proteins from different species were determined using indirect non-competitive ELISA. Antigenic components of porcine blood were determined using Western blot. The sensitivity of ELISA has been determined to analyze fish surimi that spiked with porcine plasma. After several screening of hybridoma was made, 27 hybridomas produced mAbs were selected. Based on that, fifteen mAbs are specific to raw and heated porcine blood, one mAb only specific to raw porcine blood and another 11 mAbs are cross-reacted with other animal blood. The fifteen mAbs specific to porcine blood are also not cross-reacted with meat and non-meat proteins. Based on specificity to animal plasma, twelve mAbs from 15 mAbs are specific to porcine plasma while other 3 mAbs are cross-reacted to chicken plasma. Western blot showed that 2 mAbs bind 60 kDa, 8 mAbs bind 85 kDa and 2 mAbs bind 250 kDa of the antigenic protein of porcine blood. The mAb labeled as B4E1 was selected to be used for detection of porcine plasma. The developed ELISA has a limit of detection (LOD) and limit of quantification (LOQ) of 0.2 μg/g and 1 μg/g, respectively for a standard solution of porcine plasma. The intra- and inter-assays of ELISA with coefficients of variation (CVs) less than 20% were able to detect at least 0.25% (w/w) porcine plasma in fish surimi. In conclusion, this study has successfully obtained the hybridoma-producing mAbs that are specific to porcine blood and porcine plasma. This study also has developed indirect non-competitive ELISA for detection of porcine plasma in laboratory model fish ball and fish surimi products using mAb B4E1 with LOD and LOQ of 0.2 μg/g and 1 μg/g, respectively with the sensitivity of the ELISA is 0.25% (w/w) porcine plasma in fish surimi

Comparison of bovine and porcine skin gelatin based on amino acid composition, polypeptide pattern and gel strength

The ability to compare bovine skin gelatin (BSG) and porcine skin gelatin (PSG) based on their amino acid content, polypeptide pattern and Bloom strength were investigated. Amino acid composition analysis of the two types of gelatin showed that the content of glycine, proline and arginine in PSG were higher than BSG. In the other hand, the Bloom strength of PSG and BSG was determined in different pH value ranging from pH 3 to pH 10. Within this pH range, PSG confer higher Bloom strength as compared to BSG. Both gelatins have the highest Bloom strength at pH 9 while the lowest Bloom strength of BSG and PSG was at pH 3 and pH 5 respectively. However, there is no difference of polypeptide pattern between PSG and BSG as both of them consists of α and β chains

Method for extraction and identification of gelatin

The present invention generally relates to extraction and identification of gelatin. More particularly, the present invention relates to a method of extracting gelatin protein from collagen containing raw materials and identifying the source of the extracted gelatin protein, the method comprising the steps of extracting gelatin proteins using acetone solution and separating the extracted gelatin proteins to their molecular weights by way of gel electrophoresis, wherein the molecular weight difference of the proteins is used as a biomarker to identify the source of the extracted gelatin proteins

Use of principal component analysis for differentiation of gelatine sources based on polypeptide molecular weights

The study was aimed to differentiate between porcine and bovine gelatines in adulterated samples by utilising sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE) combined with principal component analysis (PCA). The distinct polypeptide patterns of 6 porcine type A and 6 bovine type B gelatines at molecular weight ranged from 50 to 220 kDa were studied. Experimental samples of raw gelatine were prepared by adding porcine gelatine in a proportion ranging from 5% to 50% (v/v) to bovine gelatine and vice versa. The method used was able to detect 5% porcine gelatine added to the bovine gelatine. There were no differences in the electrophoretic profiles of the jelly samples when the proteins were extracted with an acetone precipitation method. The simple approach employing SDS–PAGE and PCA reported in this paper may provide a useful tool for food authenticity issues concerning gelatine

Current analytical methods for porcine identification in meat and meat products

Authentication of meat products is critical in the food industry. Meat adulteration may lead to religious apprehensions, financial gain and food-toxicities such as meat allergies. Thus, empirical validation of the quality and constituents of meat is paramount. Various analytical methods often based on protein or DNA measurements are utilized to identify meat species. Protein-based methods, including electrophoretic and immunological techniques, are at times unsuitable for discriminating closely related species. Most of these methods have been replaced by more accurate and sensitive detection methods, such as DNA-based techniques. Emerging technologies like DNA barcoding and mass spectrometry are still in their infancy when it comes to their utilization in meat detection. Gold nanobiosensors have shown some promise in this regard. However, its applicability in small scale industries is distant. This article comprehensively reviews the recent developments in the field of analytical methods used for porcine identification