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Duplex droplet digital PCR (ddPCR) for simultaneous quantification of bovine and porcine gelatin in capsules

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Abstract

Detection of bovine and porcine in gelatin-based products is important as species fraud and product mislabeling may have a detrimental impact on customers who have health, ethical, and religious concerns about animal products. The duplex droplet digital PCR (ddPCR) assay using double-quenched probes has been developed for quantification and detection of porcine and bovine DNA in gelatin capsules. A DNA mixture derived from gelatin was found to have a limit of detection as low as 0.001 ng/µl for porcine samples and 0.01 ng/µl for bovine samples. DNA from 12 other distinct species was tested with the bovine and porcine probes, showing high specificity for this method. The test was validated using fifty-five commercial supplement and pharmaceutical capsules, of which 17 were positive for bovine and/or porcine DNA. This study shows that the duplex ddPCR is reliable for routine analysis in the identification of bovine and porcine origins for gelatin capsules.

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Acknowledgements

This work was officially supported by a research grant from The Halal Science Center, Chulalongkorn University.

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Authors and Affiliations

  1. The Halal Science Center, Chulalongkorn University, CU Research Building, 254 Phayathai Road, Wangmai, Pathumwan, Bangkok, 10330, Thailand

    Pornpimol Mahamad, Winai Dahlan, Saveeyah Kahong, Sukanya So-audon, Wila Munaowaroh & Vanida Nopponpunth

  2. Department of Clinical Chemistry, Faculty of Allied Health Science, Chulalongkorn University, 154 Rama I Road, Chula Soi 12, Pathumwan, Bangkok, 10330, Thailand

    Vanida Nopponpunth

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  1. Pornpimol Mahamad
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  2. Winai Dahlan
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  3. Saveeyah Kahong
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  4. Sukanya So-audon
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  5. Wila Munaowaroh
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  6. Vanida Nopponpunth
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Contributions

PM: Experimental design, Methodology, Data collection and analysis, Manuscript preparation and revision; WD: Conceptualization, Supervision and Funding acquisition, Manuscript preparation and revision; SK: Literature review, Methodology, Manuscript preparation and revision; SS: Methodology, Data collection and analysis, Manuscript preparation and revision; WM: Data collection and analysis, Manuscript preparation and revision; VN: Contributed to experimental design, Supervision, Manuscript preparation and revision.

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Correspondence to Pornpimol Mahamad.

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Mahamad, P., Dahlan, W., Kahong, S. et al. Duplex droplet digital PCR (ddPCR) for simultaneous quantification of bovine and porcine gelatin in capsules. Food Sci Biotechnol 32, 803–811 (2023). https://doi.org/10.1007/s10068-022-01204-x

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  • DOI: https://doi.org/10.1007/s10068-022-01204-x

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