DNA and Protein-Based Methods for Halal Gelatin Authentication: A Review
DOI:
10.29303/jpm.v21i2.11539Published:
2026-04-30Downloads
Abstract
Tracing and tracking halal has emerged as an imperative for Muslim consumers worldwide to ensure that the meat-based products comply with Islamic law and food safety standards. The most challenging technical issue in halal authentication concerns products derived from gelatin. Because extreme manufacturing processes, such as high-temperature and chemical processing, damage the physical and molecular architecture of raw materials, detection is difficult. Objective: This study builds on a systematic literature review with a PRISMA protocol to map DNA- and protein-based detection techniques and compare their efficiencies. Methods: A systematic search was conducted across major databases, including Google Scholar, ScienceDirect, and ResearchGate, for peer-reviewed articles published from 2015 to 2025. From this process, 25 studies with high-impact potential were selected for thematic analysis. The results show that DNA-based methods dominate the literature (80%). Despite this, quantitative PCR (qPCR) remains the "gold standard" owing to its specificity and sensitivity. However, the findings reveal that droplet digital PCR (ddPCR) can detect highly degraded DNA in complex matrices, whereas traditional methods have shown limitations for detecting DNA in pharmaceutical capsules and cosmetics. On the other hand, protein-based methods such as ELISA remain useful for fast, economical screening, but sensitivity is substantially reduced at higher processing temperatures. In conclusion, this study highlights the importance of combining IPC and multi-method frameworks to limit false negatives. The findings of this study are substantial enough to provide the fundamentals for the alternative scaling-up of halal assurance systems, especially in the form of guidelines that provide a scientific framework for strengthening a more reliable authentication system for processed gelatin products, at least within Indonesia.
Keywords:
ddPCR; Gelatin; Halal Authentication; Porcine Detection; qPCRReferences
[1] M. A. S. Ahmad, S. S. Shukur, and N. M. Noh, “Halal integrity and authenticity in food industry: A review of issues and challenges,” J. Halal Serv., vol. 3, no. 1, pp. 15–28, 2023.
[2] W. Anggraini, “Development of biodegradable materials for consumer product packaging: Environmental impact and consumer preferences,” J. Pijar MIPA, vol. 20, no. 2, pp. 304–310, 2025, doi: 10.29303/jpm.v20i2.8588.
[3] N. S. A. Gani, S. S. Shukur, and H. Desa, “Porcine DNA authentication in sugar confectionery products,” Malaysian J. Appl. Sci., vol. 10, no. 2, pp. 31–44, 2025, doi: 10.37231/myjas.2025.10.2.476.
[4] R. M. C. Sari et al., “Recent advances in rapid and reliable biosensors for detection of porcine gelatine,” Int. J. Food Sci. Technol., vol. 59, no. 10, pp. 7716–7726, 2024, doi: 10.1111/ijfs.17129.
[5] N. Othman et al., “Molecular analysis for halal authentication of gelatin in pharmaceutical capsules,” J. Pharm. Sci., vol. 12, no. 1, pp. 45–52, 2024, doi: 10.1016/j.xphs.2023.09.012.
[6] M. T. J. Abdullah, “DNA-based and protein-based methods for porcine detection: A comparative review,” Food Chem. Res., vol. 8, no. 4, pp. 210–225, 2023.
[7] Heryani, S. Nurjanah, and D. N. Faridah, “Validation of porcine DNA analysis method using real-time PCR,” Halal Stud. Soc., vol. 1, no. 3, pp. 5–11, 2024, doi: 10.29244/hass.1.3.5-11.
[8] N. Darmawan et al., “Droplet digital PCR (ddPCR) for halal detection of commercial gelatin products,” Int. J. Food Sci., vol. 2025, no. 1, 2025, doi: 10.1155/ijfo/8841767.
[9] S. Rahmawati and A. Malik, “Limitations of ELISA in highly processed food matrices,” Indones. J. Chem. Anal., vol. 6, no. 2, pp. 88–95, 2024, doi: 10.20885/ijca.vol6.iss2.art2.
[10] M. J. Page et al., “The PRISMA 2020 statement: An updated guideline for reporting systematic reviews,” BMJ, vol. 372, p. n71, 2021, doi: 10.1136/bmj.n71.
[11] A. W. Harzing, “Publish or Perish 8,” 2021. [Online]. Available: https://harzing.com/resources/publish-or-perish
[12] V. Braun and V. Clarke, “Using thematic analysis in psychology,” Qual. Res. Psychol., vol. 3, no. 2, pp. 77–101, 2006, doi: 10.1191/1478088706qp063oa.
[13] Sudjadi et al., “Optimization of DNA extraction for porcine detection in cosmetic creams,” J. Cosmet. Sci. Technol., vol. 18, no. 2, pp. 112–120, 2020.
[14] M. Ali et al., “Multiplex PCR for simultaneous detection of bovine, porcine, and chicken DNA in food products,” Food Control, vol. 121, p. 107655, 2021.
[15] A. Fitri et al., “Validation of cytochrome b primers for porcine DNA detection in marshmallows,” Halal Sci. J., vol. 5, no. 1, pp. 22–30, 2022.
[16] S. Nurani et al., “Combination of FTIR and PCR for species-specific detection,” J. Pharm. Anal., vol. 9, no. 4, pp. 245–252, 2019, doi: 10.1016/j.jpha.2019.03.003.
[17] H. Kim et al., “Porcine-specific peptide marker identification via HPLC-MS/MS proteomics,” J. Agric. Food Chem., vol. 66, no. 15, pp. 3900–3908, 2018.
[18] A. Rohman et al., “Authenticity of fish oil capsules from porcine gelatin contamination using qPCR,” Int. J. Food Prop., vol. 20, no. 3, pp. 671–679, 2017.
[19] R. Hassan et al., “Absolute quantification of porcine DNA using digital PCR,” Food Res. Int., vol. 152, p. 110901, 2022, doi: 10.1016/j.foodres.2021.110901.
[20] Y. Erwanto et al., “PCR-RFLP for species differentiation in collagen-derived products,” Meat Sci., vol. 114, pp. 12–18, 2016, doi: 10.1016/j.meatsci.2015.12.001.
[21] M. M. Sahilah et al., “Halal monitoring of vitamin capsules,” J. Food Saf. Qual., vol. 7, no. 2, pp. 104–112, 2021.
[22] S. Maryam et al., “DNA degradation of porcine in jelly products,” Food Biosci., vol. 51, p. 102234, 2023.
[23] N. A. M. Fadzlillah et al., “Effectiveness of ELISA kits for porcine gelatin detection,” Food Anal. Methods, vol. 8, no. 5, pp. 1311–1318, 2015.
[24] D. Raraswati et al., “Low concentration DNA detection using digital PCR,” Cosmet. J., vol. 14, no. 1, pp. 45–53, 2022, doi: 10.2147/CCIDE.S346743.
[25] S. M. Yusop et al., “Phylogenetic analysis for gelatin origin confirmation,” J. Trop. Med., vol. 2020, 2020.
[26] R. Handayani et al., “Effect of cooking spices inhibitors on porcine DNA amplification,” J. Food Qual. Hazards Control, vol. 11, no. 2, pp. 78–85, 2024.
[27] N. Zakaria et al., “Shift from conventional to digital methods for halal certification,” Trends Food Sci. Technol., vol. 109, pp. 15–25, 2021, doi: 10.1016/j.tifs.2021.01.019.
[28] B. Pratama et al., “Importance of laboratory method validation in halal assurance,” Indones. J. Halal Res., vol. 5, no. 3, pp. 201–215, 2023.
[29] A. Sasmita et al., “Portable protein-based biosensor for porcine gelatin detection,” Sensors Actuators B Chem., vol. 420, p. 136502, 2025, doi: 10.1016/j.snb.2024.136502.
[30] F. Abdullah, “Comparative review of DNA vs protein-based methods for halal authentication,” Halal Res. Technol., vol. 8, no. 2, pp. 88–101, 2023.
[31] K. Heryani et al., “Internal positive control (IPC) in qPCR to prevent false negatives,” J. Food Qual., vol. 2024, 2024.
[32] L. Handayani, “Inhibitory effects of cooking spices on porcine DNA amplification,” J. Halal Res., vol. 6, no. 1, pp. 40–48, 2024.
[33] W. Anggraini, “Role of microbial enzymes in organic waste bioconversion,” J. Pijar MIPA, vol. 20, no. 5, pp. 835–841, 2025, doi: 10.29303/jpm.v20i5.9343.
[34] W. Anggraini, “Synthetic enzyme engineering for halal food applications,” J. Pijar MIPA, vol. 20, no. 7, pp. 1259–1263, 2025, doi: 10.29303/jpm.v20i7.9909.
[35] M. Ahmad et al., “Halal integrity and food fraud in gelatin supply chain,” J. Halal Sci., vol. 5, no. 2, pp. 112–125, 2023.
[36] N. Othman et al., “Molecular analysis for halal authentication of gelatin in pharmaceutical capsules,” J. Pharm. Sci., vol. 12, no. 1, pp. 45–52, 2024.
[37] N. S. A. Gani, S. S. Shukur, and H. Desa, “Porcine DNA authentication in sugar confectionery products,” Malaysian J. Appl. Sci., vol. 10, no. 2, pp. 31–44, 2025, doi: 10.37231/myjas.2025.10.2.476.
[38] A. Fitri et al., “Validation of cytochrome b primers for porcine DNA detection in high-sugar marshmallows,” Halal Sci. J., vol. 5, no. 1, pp. 22–30, 2022.
[39] Heryani, S. Nurjanah, and D. N. Faridah, “Validation of porcine DNA analysis method using selected primer and exogenous internal positive control in real-time PCR,” Halal Stud. Soc., vol. 1, no. 3, pp. 5–11, 2024, doi: 10.29244/hass.1.3.5-11.
[40] B. Pratama et al., “Importance of laboratory method validation to support halal product assurance regulations in Indonesia,” Indones. J. Halal Res., vol. 5, no. 3, pp. 201–215, 2023.
[41] K. Heryani et al., “Internal positive control (IPC) in qPCR to prevent false negatives in complex food matrices,” J. Food Qual., vol. 2024, 2024.
[42] L. Handayani, “Inhibitory effects of cooking spices on porcine DNA amplification in processed meats,” J. Halal Res., vol. 6, no. 1, pp. 40–48, 2024.
[43] R. Hassan et al., “Absolute quantification of porcine DNA in binary gelatin mixtures using digital PCR,” Food Res. Int., vol. 152, p. 110901, 2022, doi: 10.1016/j.foodres.2021.110901.
[44] S. Maryam et al., “DNA degradation of porcine in jelly products: Effect of acidic pH,” Food Biosci., vol. 51, p. 102234, 2023.
[45] D. Raraswati et al., “Low concentration DNA detection in liquid cosmetic serums using digital PCR,” Cosmet. J., vol. 14, no. 1, pp. 45–53, 2022, doi: 10.2147/CCIDE.S346743.
[46] N. Zakaria et al., “The shift from conventional to digital methods for halal certification: A global trend review,” Trends Food Sci. Technol., vol. 109, pp. 15–25, 2021, doi: 10.1016/j.tifs.2021.01.019.
[47] R. M. C. Sari et al., “Recent advances in rapid and reliable biosensors for detection of porcine gelatine,” Int. J. Food Sci. Technol., vol. 59, no. 10, pp. 7716–7726, 2024, doi: 10.1111/ijfs.17129.
[48] S. Rahmawati and A. Malik, “Limitations of ELISA in highly processed food matrices: A study on gelatin-based products,” Indones. J. Chem. Anal., vol. 6, no. 2, pp. 88–95, 2024, doi: 10.20885/ijca.vol6.iss2.art2.
[49] N. A. M. Fadzlillah et al., “Effectiveness of commercial ELISA kits for porcine gelatin detection in instant beverage powders,” Food Anal. Methods, vol. 8, no. 5, pp. 1311–1318, 2015.
[50] S. Nurani et al., “Combination of FTIR and PCR for species-specific detection in pharmaceutical capsules,” J. Pharm. Anal., vol. 9, no. 4, pp. 245–252, 2019, doi: 10.1016/j.jpha.2019.03.003.
[51] V. Braun and V. Clarke, “Using thematic analysis in psychology,” Qual. Res. Psychol., vol. 3, no. 2, pp. 77–101, 2006, doi: 10.1191/1478088706qp063oa.
[52] A. Sasmita et al., “Portable protein-based biosensor for porcine gelatin detection in hygiene products,” Sensors Actuators B Chem., vol. 420, p. 136502, 2025, doi: 10.1016/j.snb.2024.136502.
[53] W. Anggraini, “Synthetic enzyme engineering: In silico and in vivo approaches for applications in the halal food industry,” J. Pijar MIPA, vol. 20, no. 7, pp. 1259–1263, 2025, doi: 10.29303/jpm.v20i7.9909.
[54] M. Ahmad et al., “Halal integrity and food fraud in the global gelatin supply chain: A review,” J. Halal Sci., vol. 5, no. 2, pp. 112–125, 2023.
[55] F. Abdullah, “Comparative review of DNA vs protein-based methods for halal authentication,” Halal Res. Technol., vol. 8, no. 2, pp. 88–101, 2023.
[56] N. Darmawan et al., “Droplet digital PCR (ddPCR) for halal detection of commercial gelatin products,” Int. J. Food Sci., vol. 2025, no. 1, 2025, doi: 10.1155/ijfo/8841767.
[57] M. Ali et al., “Multiplex PCR for simultaneous detection of bovine, porcine, and chicken DNA in food products,” Food Control, vol. 121, p. 107655, 2021.
[58] N. S. A. Gani, S. S. Shukur, and H. Desa, “Porcine DNA authentication in sugar confectionery products,” Malaysian J. Appl. Sci., vol. 10, no. 2, pp. 31–44, 2025.
[59] A. Fitri et al., “Validation of cytochrome b primers for porcine DNA detection in high-sugar marshmallows,” Halal Sci. J., vol. 5, no. 1, pp. 22–30, 2022.
[60] Heryani, S. Nurjanah, and D. N. Faridah, “Validation of porcine DNA analysis method using selected primer and exogenous internal positive control in real-time PCR,” Halal Stud. Soc., vol. 1, no. 3, pp. 5–11, 2024.
[61] B. Pratama et al., “Importance of laboratory method validation to support halal product assurance regulations in Indonesia,” Indones. J. Halal Res., vol. 5, no. 3, pp. 201–215, 2023.
[62] K. Heryani et al., “Internal positive control (IPC) in qPCR to prevent false negatives in complex food matrices,” J. Food Qual., vol. 2024, 2024.
[63] L. Handayani, “Inhibitory effects of cooking spices on porcine DNA amplification in processed meats,” J. Halal Res., vol. 6, no. 1, pp. 40–48, 2024.
License
Copyright (c) 2026 Nurul Azizah, Welly Anggraini
This work is licensed under a Creative Commons Attribution 4.0 International License.
The following terms apply to authors who publish in this journal:
1. Authors retain copyright and grant the journal first publication rights, with the work simultaneously licensed under a Creative Commons Attribution License 4.0 International License (CC-BY License) that allows others to share the work with an acknowledgment of the work's authorship and first publication in this journal.
2. Authors may enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., posting it to an institutional repository or publishing it in a book), acknowledging its initial publication in this journal.
3. Before and during the submission process, authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website), as this can lead to productive exchanges as well as earlier and greater citation of published work (See The Effect of Open Access).
Downloads
How to Cite
