Novel multiplex PCR-RFLP assay discriminates bovine, porcine and fish gelatin substitution in Asian pharmaceuticals capsule shells
Gelatin is widely used in pharmaceuticals as a protective coating, such as soft and hard capsule shells. However, the animal source of gelatin is a sensitive issue because certain gelatins such as porcine and bovine gelatins are not welcome in Halal, Kosher and Hindus’ consumer goods. Recently, we h...
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| Format: | Article |
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Taylor & Francis
2018
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| author | Sultana, Sharmin Hossain, M.A Motalib Naquiah, Nina Nizar Ahmad Ali, Md. Eaqub |
| author_facet | Sultana, Sharmin Hossain, M.A Motalib Naquiah, Nina Nizar Ahmad Ali, Md. Eaqub |
| author_sort | Sultana, Sharmin |
| collection | UM |
| description | Gelatin is widely used in pharmaceuticals as a protective coating, such as soft and hard capsule shells. However, the animal source of gelatin is a sensitive issue because certain gelatins such as porcine and bovine gelatins are not welcome in Halal, Kosher and Hindus’ consumer goods. Recently, we have documented DNA barcoding and multiplex PCR platforms for discriminating porcine, bovine and fish gelatins in various fish and confectionary products; but those assays were not self-authenticating and also not tested in highly refined pharmaceutical products. To address this knowledge gap, here we report a self-authenticating multiplex PCR-restriction fragment length polymorphism (RFLP) assay to identify animal sources of various gelatin in pharmaceutical capsules. Three different restriction enzymes, BsaAI, Hpy188I and BcoDI were used to yield distinctive RFLP patterns for gelatin-based bovine (26, 94 bp), fish (97, 198 bp) and porcine (17, 70 bp) DNA in control experiments. The specificity was cross-tested against 16 non-target species and the optimised assay was used to screen gelatin sources in 30 halal-branded pharmaceuticals capsule shells. Bovine and porcine DNA was found in 27 and 3 of the 30 different capsules products. The assay was suitable for detecting 0.1 to 0.01 ng total DNA extracted from pure and mixed gelatins. The study might be useful to authenticate and monitor halal, kosher, vegetarian and Hindu compliant pharmaceuticals, foods and cosmetics. |
| first_indexed | 2024-03-06T05:51:40Z |
| format | Article |
| id | um.eprints-20593 |
| institution | Universiti Malaya |
| last_indexed | 2024-03-06T05:51:40Z |
| publishDate | 2018 |
| publisher | Taylor & Francis |
| record_format | dspace |
| spelling | um.eprints-205932019-03-05T01:31:01Z http://eprints.um.edu.my/20593/ Novel multiplex PCR-RFLP assay discriminates bovine, porcine and fish gelatin substitution in Asian pharmaceuticals capsule shells Sultana, Sharmin Hossain, M.A Motalib Naquiah, Nina Nizar Ahmad Ali, Md. Eaqub Q Science (General) QD Chemistry Gelatin is widely used in pharmaceuticals as a protective coating, such as soft and hard capsule shells. However, the animal source of gelatin is a sensitive issue because certain gelatins such as porcine and bovine gelatins are not welcome in Halal, Kosher and Hindus’ consumer goods. Recently, we have documented DNA barcoding and multiplex PCR platforms for discriminating porcine, bovine and fish gelatins in various fish and confectionary products; but those assays were not self-authenticating and also not tested in highly refined pharmaceutical products. To address this knowledge gap, here we report a self-authenticating multiplex PCR-restriction fragment length polymorphism (RFLP) assay to identify animal sources of various gelatin in pharmaceutical capsules. Three different restriction enzymes, BsaAI, Hpy188I and BcoDI were used to yield distinctive RFLP patterns for gelatin-based bovine (26, 94 bp), fish (97, 198 bp) and porcine (17, 70 bp) DNA in control experiments. The specificity was cross-tested against 16 non-target species and the optimised assay was used to screen gelatin sources in 30 halal-branded pharmaceuticals capsule shells. Bovine and porcine DNA was found in 27 and 3 of the 30 different capsules products. The assay was suitable for detecting 0.1 to 0.01 ng total DNA extracted from pure and mixed gelatins. The study might be useful to authenticate and monitor halal, kosher, vegetarian and Hindu compliant pharmaceuticals, foods and cosmetics. Taylor & Francis 2018 Article PeerReviewed Sultana, Sharmin and Hossain, M.A Motalib and Naquiah, Nina Nizar Ahmad and Ali, Md. Eaqub (2018) Novel multiplex PCR-RFLP assay discriminates bovine, porcine and fish gelatin substitution in Asian pharmaceuticals capsule shells. Food Additives & Contaminants: Part A, 35 (9). pp. 1662-1673. ISSN 1944-0049, DOI https://doi.org/10.1080/19440049.2018.1500719 <https://doi.org/10.1080/19440049.2018.1500719>. https://doi.org/10.1080/19440049.2018.1500719 doi:10.1080/19440049.2018.1500719 |
| spellingShingle | Q Science (General) QD Chemistry Sultana, Sharmin Hossain, M.A Motalib Naquiah, Nina Nizar Ahmad Ali, Md. Eaqub Novel multiplex PCR-RFLP assay discriminates bovine, porcine and fish gelatin substitution in Asian pharmaceuticals capsule shells |
| title | Novel multiplex PCR-RFLP assay discriminates bovine, porcine and fish gelatin substitution in Asian pharmaceuticals capsule shells |
| title_full | Novel multiplex PCR-RFLP assay discriminates bovine, porcine and fish gelatin substitution in Asian pharmaceuticals capsule shells |
| title_fullStr | Novel multiplex PCR-RFLP assay discriminates bovine, porcine and fish gelatin substitution in Asian pharmaceuticals capsule shells |
| title_full_unstemmed | Novel multiplex PCR-RFLP assay discriminates bovine, porcine and fish gelatin substitution in Asian pharmaceuticals capsule shells |
| title_short | Novel multiplex PCR-RFLP assay discriminates bovine, porcine and fish gelatin substitution in Asian pharmaceuticals capsule shells |
| title_sort | novel multiplex pcr rflp assay discriminates bovine porcine and fish gelatin substitution in asian pharmaceuticals capsule shells |
| topic | Q Science (General) QD Chemistry |
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