DIFFERENTIATION OF BOVINE AND PORCINE GELATINES USING LC-MS/MS AND CHEMOMETRICS

  • NINA SALAMAH Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Universitas Gadjah Mada, Yogyakarta, 55281, Indonesia
  • YUNY ERWANTO Faculty of Animal Sciences, Universitas Gadjah Mada, Yogyakarta 55281, Indonesia
  • SUDIBYO MARTONO Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Universitas Gadjah Mada, Yogyakarta, 55281, Indonesia
  • IRFAN MAULANA MS Senior Application Chemist, PT. Kromtekindo Utama “Waters Indonesia” Jakarta 12330, Indonesia
  • ABDUL ROHMAN Institute of Halal Industry and Systems, Universitas Gadjah Mada, Yogyakarta, 55281, Indonesia

Abstract

Objective: To differentiate porcine gelatin and bovine gelatins using specific peptide markers as determined by liquid chromatography-mass spectrometry tandem with mass spectrometry (LC-MS/MS) and classify both gelatins using retention time and m/z as variables in principal component analysis (PCA).


Methods: Porcine and bovine gelatins were digested using trypsin enzyme and then subjected to LC-MS/MS analysis. The specific peptides were identified. The classification between porcine and bovine gelatins was carried out using chemometrics of PCA using retention times and mass to charge ratio (m/z) as variables.


Results: PCA using singvariables retention times (tR) and m/z could successfully classify porcine gelatin and bovine gelatin based on score plots of first principle component (PC1) and second principle component (PC2). The loading plot analysis showed that variable of tR32 and m/z32 contributed for the separation of both gelatins.


Conclusion: The chromatograms of LC-MS/MS combined with PCA offered reliable method for differentiation between porcine and bovine gelatins. The developed method could be extended for halal authentication of food and pharmaceutical products via detection of porcine gelatin, non-halal gelatin.

Keywords: LC-MS/MS, Porcine gelatin, Bovine gelatin, Principal component analysis, Classification

References

1. Li GY, Fukunaga S, Takenouchi K. Comparative study of the physiological properties of collagen, gelatin and collagen hydrolysate as cosmetic materials. Int J Cosmet Sci 2005;27:101–6.
2. Poppe J, In: A Immeson. Ed. Gelatin in Thickening and Gelling Agents for Food, Blackie, London; 1992. p. 98–123.
3. Shoshani D, Markovitz E, Cohen Y. Skin test hypersensitivity study of a crosslinked, porcine collagen implant for aesthetic surgery. J Periodontol 2007;78:112–21.
4. Gómez Guillen MC, Gimenez B, Lopez Caballero ME, Montero MP. Functional and bioactive properties of collagen and gelatin from alternative sources: a review. Food Hydrocolloids 2011;25:1813-27.
5. Mursyidi A. The role of chemical analysis in the halal authentication of food and pharmaceutical products. J Food Pharm Sci 2013;1:1-4.
6. Widyaninggar A, Triyana K, Rohman A. Differentiation between porcine and bovine gelatin in capsule shells based on amino acid profiles and principal component analysis. Indonesian J Pharm 2012;23:104-9.
7. Sakaguchi M, Nakayama T, Inouye S. Food allergy to gelatin in children with systemic immediate-type reactions, including anaphylaxis, to vaccines. J Allergy Clin Immunol 1996;98:1058–61.
8. Nhari RMHR, Ismail A, Che Man YB. Analytical methods for gelatin differentiation from bovine and porcine origins and food products. J Food Sci 2012;77:R42-6.
9. Rohman A, Che Man YB. Analysis of pig derivatives for halal authentication studies. Food Rev Int 2012;28:97-112.
10. Ali ME, Sultana S, Hamid SBA, Hossain M, Yehya WA, Kader A, et al. Gelatin controversies in food, pharmaceuticals, and personal care products: authentication methods, current status, and future challenges. Crit Rev Food Sci Nutr 2016;58:1-17.
11. Venien A, Levieux D. Differentiation of bovine from porcine gelatines using polyclonal anti-peptide antibodies in indirect and competitive indirect ELISA. J Pharm Biomed Anal 2005;39:418–24.
12. Hashim DM, Che Man YB, Norakasha R, Shuhaimi M, Salmah Y, Syahariza ZA. Potential use of fourier transform infrared spectroscopy for differentiation of bovine and porcine gelatins. Food Chem 2010;118:856-60.
13. Kauschke SG, Knorr A, Heke M. Two assays for measuring fibrosis: reverse transcriptase-polymerase chain reaction of collagen alpha (III) mRNA is an early predictor of subsequent collagen deposition while a novel serum Nterminal procollagen (III) propeptide assay reflects manifest fibrosis in carbon tetrachloride-treated rats. Anal Biochem 1999;275:131–40.
14. Sudjadi, 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;19:2127-34.
15. Nemati M, Oveisi MR, Abdollahi H, Sabzevari O. Differentiation of bovine and porcine gelatins using principal component analysis. J Pharm Biomed Anal 2004;34:485–92.
16. Azilawati MI, Hashim DM, Jamilah B, Amin I. RP-HPLC method using 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate incorporated with normalization technique in principal component analysis to differentiate the bovine, porcine and fish gelatins. Food Chem 2015;172:368-76.
17. Zhang GF, Liu T, Wang Q, Chen L, Lei JD, Luo J, et al. Mass spectrometric detection of marker peptides in tryptic digests of gelatin: a new method to differentiate between bovine and porcine gelatin. Food Hydrocoll 2009; 23:2001–7.
18. Zhang GF, Sun AM, Li WJ, Liu T, Su ZG. Mass spectrometric analysis of enzymatic digestion of denatured collagen for identification of collagen type. J Chromatogr A 2006; 1114:274–7.
19. Olsen JV, Ong SE, Mann M. Trypsin cleaves exclusively c-terminal to arginine and lysine residues. Mol Cell Proteomics 2014;3:608-14.
20. Cheng XL, Wei F, Xiao XY, Zhao YY, Shi Y, Liu W, et al. Identification of five gelatins by ultra performance liquid chromatography/time-of-flight mass spectrometry (UPLC/Q-TOF-MS) using principal component analysis. J Pharm Biomed Anal 2012;62:191-5.
21. Che Man YB, Rohman A, Mansor TST. Differentiation of lard from other edible fats and oils by means of fourier transform infrared spectroscopy and chemometrics. J Am Oil Chem Soc 2011;88:187–92.
22. Marina AM, Che Man YB, Ismail A. Use of the SAW sensor electronic nose for detecting the adulteration of virgin coconut oil with RBD palm kernel olein. J Am Oil Chem Soc 2010;87:263–70.
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SALAMAH, N., ERWANTO, Y., MARTONO, S., MAULANA, I., & ROHMAN, A. (2019). DIFFERENTIATION OF BOVINE AND PORCINE GELATINES USING LC-MS/MS AND CHEMOMETRICS. International Journal of Applied Pharmaceutics, 11(4), 159-163. https://doi.org/10.22159/ijap.2019v11i4.30248
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