Halal Authentication of Gelatin Capsules using Polymerase Chain Reaction, Fourier Transform Infrared Spectroscopy and Chemometric Methods
Gelatin, a high molecular weight polypeptide obtained from protein, is an important material for capsules of health supplements.A reliable method for halal authentication of gelatin is needed because some supplement products are capsulated using gelatinfrom non-halal source.In this study a real-time...
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my-usim-ddms-127102024-05-29T19:04:58Z Halal Authentication of Gelatin Capsules using Polymerase Chain Reaction, Fourier Transform Infrared Spectroscopy and Chemometric Methods Ahlam Inayatullah Gelatin, a high molecular weight polypeptide obtained from protein, is an important material for capsules of health supplements.A reliable method for halal authentication of gelatin is needed because some supplement products are capsulated using gelatinfrom non-halal source.In this study a real-time polymerase chain reaction (PCR) assay using SYBR green fluorescent dye was done to detect porcine in gelatin capsules supplements. The method combined the porcine and bovine specific primers and SYBR green dye to specifically amplify a 120-bp fragment of bovine and 131-bp fragment of porcine mitochondrial Cytochrome b (CYT b) gene.Mitochondrial genes were present in multiple copies and thus ensure available targets even in degraded samples. Using basic local alignment search tools (BLAST) with 1 ng DNA standard, bovine and porcine species yielded a melt temperature at ±80.00 °C only with one specific peak in the melt curve reaction, demonstrating the porcine and bovine specificity of the primers. A sensitivity test with 5-fold serial dilution revealed that the assay can determine 10¯¹ nġ/ų1 of porcine and bovine DNA with efficiencies of 97.4% and 90.6% with good repeatabilities and high correlation coefficients (R²= 0.995 bovine; R²= 0.962 porcine).From a total of 20 samples of supplements in capsules with halal certifications from various countries,eight (8) samples were confirmed with adulteration of porcine DNA while four samples were confirmed to contain bovine DNA.FTIR technique was then used to confirm the results of real-time PCR in terms of chemical representation. The FTIR results were later confirmed using principal component analysis (PCA).Eight wavelengths chosen from the twelve samples were 694,1249,1342,1404,1450,1558,1635 and 3271 cm¯¹. Universiti Sains Islam Malaysia 2018 Thesis en https://oarep.usim.edu.my/handle/123456789/12710 https://oarep.usim.edu.my/bitstreams/2a995984-b079-4b92-9bf8-d1317e62ab3a/download 8a4605be74aa9ea9d79846c1fba20a33 https://oarep.usim.edu.my/bitstreams/7caf5bac-d47a-4b19-898a-d7be309a3065/download 7de1fb8f6548fbab5b6217f854f531fa https://oarep.usim.edu.my/bitstreams/c28dd736-36a2-46d8-97bd-93d4cf93315b/download 8f4cb22a71dc8047849c167cac1c44c9 https://oarep.usim.edu.my/bitstreams/6b00ee92-0ae9-4090-97e9-cf50f429b045/download dd0ae903dc776c02659d7635b1a7dc01 https://oarep.usim.edu.my/bitstreams/3e91b87c-4e0b-433d-a7c4-bf9de6682292/download 1b32e099fdf38d3264ae5aed833e9270 https://oarep.usim.edu.my/bitstreams/57efe39f-33ea-4644-945a-49571a1c0d24/download 26ca9993d4f89cc55661db558f26fdd5 https://oarep.usim.edu.my/bitstreams/e1fb506c-a3d8-465d-8064-e43e95aa5bd7/download 1e56254d5b8572d934bb766adbd66c1f https://oarep.usim.edu.my/bitstreams/41f2fd22-75ea-4aa2-abfe-151bf44661b5/download 9e72e2b2d3257ddf6013aa0ae53143da https://oarep.usim.edu.my/bitstreams/b839818d-0679-4ea5-be4e-5d72f88bf0ea/download 796b47012657f67c0a6fd7d0427d844f https://oarep.usim.edu.my/bitstreams/77e97a98-e72b-4f5a-9092-261cb39ccd2e/download 68b329da9893e34099c7d8ad5cb9c940 https://oarep.usim.edu.my/bitstreams/ce6eff6c-0e29-4258-adf9-5d80a1c32982/download 36c6f0b2061da514c400c0bc2749b5cf https://oarep.usim.edu.my/bitstreams/407b7312-6e4e-4882-82c3-f7550d16679f/download cc9067c2ee470dc248b14b194209a34e https://oarep.usim.edu.my/bitstreams/26adaa13-36d6-4235-82f9-1240cc32c6b2/download 48175cc56e52e020bf178616c0977374 https://oarep.usim.edu.my/bitstreams/6f031891-b328-43a4-a54d-ada0339ca7c3/download 212b0306580d4f0044d18f9a3edcc832 https://oarep.usim.edu.my/bitstreams/9d9d71ca-0f3f-4faa-91a3-69aa0df44938/download 792680ea1ef80bbcb0718ee7aa1e2a75 https://oarep.usim.edu.my/bitstreams/306f7679-8381-42be-adca-03d9133d48db/download e1c06d85ae7b8b032bef47e42e4c08f9 https://oarep.usim.edu.my/bitstreams/154e7eeb-e117-4c0b-8727-d9ec1de92597/download 429079e52f342c49301a02e207bfde06 Gelatin mitochondrial Cytochrome |
| institution |
Universiti Sains Islam Malaysia |
| collection |
USIM Institutional Repository |
| language |
English |
| topic |
Gelatin mitochondrial Cytochrome |
| spellingShingle |
Gelatin mitochondrial Cytochrome Ahlam Inayatullah Halal Authentication of Gelatin Capsules using Polymerase Chain Reaction, Fourier Transform Infrared Spectroscopy and Chemometric Methods |
| description |
Gelatin, a high molecular weight polypeptide obtained from protein, is an important material for capsules of health supplements.A reliable method for halal authentication of gelatin is needed because some supplement products are capsulated using gelatinfrom non-halal source.In this study a real-time polymerase chain reaction (PCR) assay using SYBR green fluorescent dye was done to detect porcine in gelatin capsules supplements. The method combined the porcine and bovine specific primers and SYBR green dye to specifically amplify a 120-bp fragment of bovine and 131-bp fragment of porcine mitochondrial Cytochrome b (CYT b) gene.Mitochondrial genes were present in multiple copies and thus ensure available targets even in degraded samples. Using basic local alignment search tools (BLAST) with 1 ng DNA standard, bovine and porcine species yielded a melt temperature at ±80.00 °C only with one specific peak in the melt curve reaction, demonstrating the porcine and bovine specificity of the primers. A sensitivity test with 5-fold serial dilution revealed that the assay can determine 10¯¹ nġ/ų1 of porcine and bovine DNA with efficiencies of 97.4% and 90.6% with good repeatabilities and high correlation coefficients (R²= 0.995 bovine; R²= 0.962 porcine).From a total of 20 samples of supplements in capsules with halal certifications from various countries,eight (8) samples were confirmed with adulteration of porcine DNA while four samples were confirmed to contain bovine DNA.FTIR technique was then used to confirm the results of real-time PCR in terms of chemical representation. The FTIR results were later confirmed using principal component analysis (PCA).Eight wavelengths chosen from the twelve samples were 694,1249,1342,1404,1450,1558,1635 and 3271 cm¯¹. |
| format |
Thesis |
| author |
Ahlam Inayatullah |
| author_facet |
Ahlam Inayatullah |
| author_sort |
Ahlam Inayatullah |
| title |
Halal Authentication of Gelatin Capsules using Polymerase Chain Reaction, Fourier Transform Infrared Spectroscopy and Chemometric Methods |
| title_short |
Halal Authentication of Gelatin Capsules using Polymerase Chain Reaction, Fourier Transform Infrared Spectroscopy and Chemometric Methods |
| title_full |
Halal Authentication of Gelatin Capsules using Polymerase Chain Reaction, Fourier Transform Infrared Spectroscopy and Chemometric Methods |
| title_fullStr |
Halal Authentication of Gelatin Capsules using Polymerase Chain Reaction, Fourier Transform Infrared Spectroscopy and Chemometric Methods |
| title_full_unstemmed |
Halal Authentication of Gelatin Capsules using Polymerase Chain Reaction, Fourier Transform Infrared Spectroscopy and Chemometric Methods |
| title_sort |
halal authentication of gelatin capsules using polymerase chain reaction, fourier transform infrared spectroscopy and chemometric methods |
| granting_institution |
Universiti Sains Islam Malaysia |
| url |
https://oarep.usim.edu.my/bitstreams/7caf5bac-d47a-4b19-898a-d7be309a3065/download https://oarep.usim.edu.my/bitstreams/c28dd736-36a2-46d8-97bd-93d4cf93315b/download https://oarep.usim.edu.my/bitstreams/6b00ee92-0ae9-4090-97e9-cf50f429b045/download https://oarep.usim.edu.my/bitstreams/3e91b87c-4e0b-433d-a7c4-bf9de6682292/download https://oarep.usim.edu.my/bitstreams/57efe39f-33ea-4644-945a-49571a1c0d24/download https://oarep.usim.edu.my/bitstreams/e1fb506c-a3d8-465d-8064-e43e95aa5bd7/download https://oarep.usim.edu.my/bitstreams/41f2fd22-75ea-4aa2-abfe-151bf44661b5/download https://oarep.usim.edu.my/bitstreams/b839818d-0679-4ea5-be4e-5d72f88bf0ea/download |
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