Determination of bovine and porcine gelatin polypeptides using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE)

Gelatin is used widely in food and pharmaceutical industries because of its unique physicochemical characteristics. However, the gelatin utilisation becomes an issue among Muslim, Jews and vegetarians due to its animal origin. As such, methods of gelatin detection and differentiation as well as gela...

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Main Author:	Tukiran, Nur Azira
Format:	Thesis
Language:	English
Published:	2012
Subjects:	Gelatin Gelatin - Analysis Polyacrylamide gel electrophoresis
Online Access:	http://psasir.upm.edu.my/id/eprint/33346/1/IPPH%202012%203R.pdf
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id	my-upm-ir.33346
record_format	uketd_dc
spelling	my-upm-ir.333462015-03-26T03:10:23Z Determination of bovine and porcine gelatin polypeptides using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) 2012-04 Tukiran, Nur Azira Gelatin is used widely in food and pharmaceutical industries because of its unique physicochemical characteristics. However, the gelatin utilisation becomes an issue among Muslim, Jews and vegetarians due to its animal origin. As such, methods of gelatin detection and differentiation as well as gelatin stability in the processed foods have emerged as an important research area to be studied. Therefore, the aim of this study was to detect and differentiate the bovine and porcine gelatin polypeptides using a combination method of sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and Principal Component Analysis (PCA). Utilising optimal conditions of SDS-PAGE with 6% of resolving gel, 8 μg amount of protein, 8 M Urea-SDS sample buffer, together with sensitive silver staining had exhibited molecular weights of gelatin polypeptides ranged from 53 to 220 kDa. They can be used to differentiate between bovine and porcine gelatins. The gelatin of porcine exhibited wider molecular weight distribution as compared to bovine that consisted of 11 and 2 prominent bands, respectively. In addition, these prominent bands were stable under heat treatment and consistent even in different Bloom number offer suitability for the identification of gelatin sources. In order to determine the detection limit of SDS-PAGE in detecting the percentage of adulterated samples,PCA was applied to classify the percentage of adulteration in the samples. Results showed that SDS-PAGE combined with PCA was unable to detect the presence of less than 5% porcine gelatin adulterated in the bovine gelatin. To detect the presence of gelatin in the samples, both extracting solutions (cold acetone and deionised water) are suitable; however, for commercial processed products with mixed ingredients cold acetone was more efficient. In the study, the qualitative comparison using SDS-PAGE between samples could be differentiated by PCA, and the combination may provide robust information for gelatin species identification. This study proposed that this new approach employing SDS-PAGE and PCA together with a simple gelatin extraction method may provide a useful tool for food authenticity issues concerning gelatin. Gelatin Gelatin - Analysis Polyacrylamide gel electrophoresis 2012-04 Thesis http://psasir.upm.edu.my/id/eprint/33346/ http://psasir.upm.edu.my/id/eprint/33346/1/IPPH%202012%203R.pdf application/pdf en public masters Universiti Putra Malaysia Gelatin Gelatin - Analysis Polyacrylamide gel electrophoresis
institution	Universiti Putra Malaysia
collection	PSAS Institutional Repository
language	English
topic	Gelatin Gelatin - Analysis Polyacrylamide gel electrophoresis
spellingShingle	Gelatin Gelatin - Analysis Polyacrylamide gel electrophoresis Tukiran, Nur Azira Determination of bovine and porcine gelatin polypeptides using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE)
description	Gelatin is used widely in food and pharmaceutical industries because of its unique physicochemical characteristics. However, the gelatin utilisation becomes an issue among Muslim, Jews and vegetarians due to its animal origin. As such, methods of gelatin detection and differentiation as well as gelatin stability in the processed foods have emerged as an important research area to be studied. Therefore, the aim of this study was to detect and differentiate the bovine and porcine gelatin polypeptides using a combination method of sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and Principal Component Analysis (PCA). Utilising optimal conditions of SDS-PAGE with 6% of resolving gel, 8 μg amount of protein, 8 M Urea-SDS sample buffer, together with sensitive silver staining had exhibited molecular weights of gelatin polypeptides ranged from 53 to 220 kDa. They can be used to differentiate between bovine and porcine gelatins. The gelatin of porcine exhibited wider molecular weight distribution as compared to bovine that consisted of 11 and 2 prominent bands, respectively. In addition, these prominent bands were stable under heat treatment and consistent even in different Bloom number offer suitability for the identification of gelatin sources. In order to determine the detection limit of SDS-PAGE in detecting the percentage of adulterated samples,PCA was applied to classify the percentage of adulteration in the samples. Results showed that SDS-PAGE combined with PCA was unable to detect the presence of less than 5% porcine gelatin adulterated in the bovine gelatin. To detect the presence of gelatin in the samples, both extracting solutions (cold acetone and deionised water) are suitable; however, for commercial processed products with mixed ingredients cold acetone was more efficient. In the study, the qualitative comparison using SDS-PAGE between samples could be differentiated by PCA, and the combination may provide robust information for gelatin species identification. This study proposed that this new approach employing SDS-PAGE and PCA together with a simple gelatin extraction method may provide a useful tool for food authenticity issues concerning gelatin.
format	Thesis
qualification_level	Master's degree
author	Tukiran, Nur Azira
author_facet	Tukiran, Nur Azira
author_sort	Tukiran, Nur Azira
title	Determination of bovine and porcine gelatin polypeptides using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE)
title_short	Determination of bovine and porcine gelatin polypeptides using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE)
title_full	Determination of bovine and porcine gelatin polypeptides using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE)
title_fullStr	Determination of bovine and porcine gelatin polypeptides using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE)
title_full_unstemmed	Determination of bovine and porcine gelatin polypeptides using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE)
title_sort	determination of bovine and porcine gelatin polypeptides using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (sds-page)
granting_institution	Universiti Putra Malaysia
publishDate	2012
url	http://psasir.upm.edu.my/id/eprint/33346/1/IPPH%202012%203R.pdf
_version_	1747811697021681664

Determination of bovine and porcine gelatin polypeptides using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE)

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