Detection of tilapia, salmon, shark and shrimp in gelatin products by PCR method for the first time in Iran

Document Type : Original Article

Authors

1 Dana Gene Pajouh Knowledgebase Company, Tehran, Iran

2 Halal Research Center of IRI, Ministry of Health and Medical Education,, Tehran, Iran

Abstract

Introduction: Gelatin is made from the collagen of the skin and bones of animals and is widely used in food, pharmaceutical, cosmetics. Given the importance of the halal status of gelatin used in these products, the production and supply of halal gelatin are important in terms of global trade. Today, the solubility of gelatinous products is controlled for the origin of pigs or cattle, but in cases where the capsules or supplements are from fish and aquatic gelatin, they are not detected.
Methods: Specific primers for tilapia, salmon, sharks and shrimp were designed using databases and Bioinformatics studies. Then the PCR method was optimized. Finally, the DNA extracted from the capsules, collagen and supplements were amplified with specific primers and displayed on agarose gel.
Results: PCR reaction with specific primers of tilapia, salmon, shark and shrimp showed 113, 122, 200 and 132 bp bands on agarose gel, respectively. The specificity, stability and repeatability of the method were confirmed for each pair of primers. PCR with DNA extracted from gelatin and collagen capsules also confirmed the origin of aquatic animals.
Conclusion: In this study, for the first time, the origin of collagen and gelatin capsules were identified using specific primers of tilapia, salmon, shark and shrimp and PCR. The results showed that specific primers are able to amplify the expected band and identify the source. Therefore, with this method, the solubility of the mentioned cases can be checked, approved or rejected in a short period of time before entering the country.

Keywords


 1. Sudjadi S, Wardani HS, Sepminarti T, Rohman A. Analysis of porcine Gelatin DNA in a commercial capsule shell using real-time polymerase chain reaction for halal authentication. Int J Food Prop. 2016, 19 (9): 2127–34.
2. Nikzad J, Shahhosseini S, Tabarzad M, Nafissi-Varcheh N, Torshabi M. Simultaneous detection of bovine and porcine DNA in pharmaceutical gelatin capsules by duplex PCR assay for Halal authentication. DARU Journal of Pharmaceutical Sciences. 2017, 25:3, DOI 10.1186/s40199-017-0171-3.
3. Sepminarti, T, Sudjadi S, Wardani HS, Rohman A. Real-time polymerase chain reaction for halal authentication of gelatin in soft candy. Asian Journal of Biochemistry. 2016, 11 (1): 34-43.
4. Erwanto, Y, Rohman A, Arsyanti L, Pranoto Y. Identification of pig DNA in food products using polymerase chain reaction (PCR) for halal authentication-a review International. Food Research Journal. 2018, 25(4): 1322-1331.
5. Raja Mohd Hafidz RN, Ismail A, Che Man YB. Analytical Methods for Gelatin Differentiation from Bovine and Porcine Origins and Food Products. Journal of Food Science. 2012, 71:1.
6. Jae-Hwang L, Mi-Ra K, Cheon-Ho J, Yoo-Kyung J, Kisung K, Tae Sun K.Specific PCR assays to determine bovine, porcine, fish and plant origin of gelatin capsules of dietary supplements. Food Chemistry. 2016, 211: 253-259. DOI, https://doi.org/10.1016/j.foodchem.2016.05.060.
7. Bakar J. Properties of gelatins from skins of fish - Black tilapia (Oreochromis mossambicus) and red tilapia (Oreochromis nilotica). Food Chemistry. 2002, 77 (1):81-84. DOI, 10.1016/S0308-8146(01)00328-4
8. Panida S, Jirarat T, Pitt S. Extraction and electrospinning of gelatin from fish skin. International Journal of Biological Macromolecules. 2008,42(3):247-55. DOI, 10.1016/j.ijbiomac.2007.11.005
9. O. Martins ME, R. Sosa J, L. Claudino R, O. Lino SC, Vale DA. Thermal and Chemical Properties of Gelatin from Tilapia (Oreochromis niloticus) Scale. Journal of Aquatic Food Product Technology.2018,27 (10): 1120-1133.
10. Jeevithan E, Bao B, Bu Y, Zhou Y, Zhao Q, Wu W. Type II Collagen and Gelatin from Silvertip Shark (Carcharhinus albimarginatus) Cartilage: Isolation, Purification, Physicochemical and Antioxidant Properties. Mar Drugs. 2014, 12 (7): 3852–3873. DOI, 10.3390/md12073852.
11. Shahiri Tabarestani H, Sedaghat N, Jahanshahi M, Motamedzadeghan A, Mohebbi M. Physicochemical and Rheological Properties of White-Cheek Shark (Carcharhinus dussumieri) Skin Gelatin. International Journal of Food Properties. 2016, 19 (12): 2788-2804.
14. Matiacevich S, Cftre DC, Schebor C, Enrione J. Physicochemical and antimicrobial properties of bovine and salmon gelatin-chitosan films. CyTA - Journal of Food. 2013, 11 (4): 366-378. 
15. Fan HY, Dumont MJ, Simpson BK. Extraction of gelatin from salmon (Salmo salar) fish skin using trypsin-aided process: optimization by Plackett–Burman and response surface methodological approaches. J Food Sci Technol. 2017, 54 (12): 4000–4008. DOI, 10.1007/s13197-017-2864-5.