Simultaneous identification and verification of gelatin type in capsule shells by electrophoresis and polymerase chain reaction

Malik, Amarila; Sutantyo, Melya Leonita; Hapsari, Indah; Sinurat, Anna Veronika; Purwati, Euis Maras; Jufri, Mahdi; Suryadi, Herman

doi:10.1007/s40005-016-0245-0

Cited by 15 publications

(14 citation statements)

References 25 publications

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“…1-A, the PCR amplification produced amplicon bands of approximately 360 bp length for both porcine and bovine. This result is consistent with the result reported for porcine and bovine amplifications using universal cyt b primer pairs [12].…”

Section: Optimization Of Pcr-rflpsupporting

confidence: 93%

“…One pair of universal primer and two pairs of porcine-specific primers were selected from the literature review, as listed in PCR-RFLP was performed as described previously by (Malik et al 2016). Amplicon DNA was subjected to digestion by using Bsa JI restriction enzyme (New England Biolabs, USA).…”

Section: Primer Characterization Analysismentioning

confidence: 99%

“…Based on in silico analysis and reports by Aida et al (2005), Malik et al (2016), and Murugaiah et al (2009) [11,12,24] the appropriate restriction enzyme to discriminate cytochrome b DNAs of porcine and bovine is BsaJI enzyme. Correct DNA fragments, i.e.…”

Section: Optimization Of Pcr-rflpmentioning

confidence: 99%

“…polymerase chain reaction (PCR) method [1]. Several PCR-based methods such as PCRsouthern hybridization [1,10], polymerase chain reactionrestriction fragment length polymorphism (PCR-RFLP) [11,12], real-time PCR [13], nested-PCR [14], and multiplex PCR [13] have widely established and proved that PCR-based analysis is sensitive and specific in identifying several species in low amount of DNA.…”

Section: Introductionmentioning

confidence: 99%

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Rapid Pcr–based Detection Optimization of Porcine Dna in Gelatin Capsule Shell

2018

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Objective: This study was conducted to optimize the genomic deoxyribonucleic acid (DNA) based molecular detection of gelatin derived from porcine by performing polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and duplex PCR method employing cyt B gene.Methods: Optimization was carried out for DNA extraction, PCR conditions, and the sensitivity of the PCR-RFLP method. Due to the very low DNA trace in gelatin after the various manufacturing process, the extraction was optimized to obtain sufficient DNA which was visible on the agarose gel. PCR-RFLP was carried out using universal primers and BsaJI restriction enzyme, and duplex PCR was carried out using two sets of porcine-specific primers. Porcine and bovine DNA were mixed in various concentration to confirm sensitivity of both methods, i.e. 100%, 50%, 10%, 1%, 0.5%, 0.1%, 0.05%, and 0.01%Results: Both methods, PCR-RFLP, and Duplex PCR, were able to detect as low as 0.01% porcine DNA, indicated by the presence of porcine DNA amplicon bands (131 bp and 228 bp for PCR-RFLP, 212 bp and 398 bp for duplex PCR). Although DNA bands presented in low intensity, identification of porcine and bovine species and estimation of DNA quantities were possible.Conclusion: Both conventional PCR methods, i.e. PCR-RFLP and Duplex PCR, were sensitive, specific, and suitable as a rapid initial detection method for molecular detection of porcine in gelatin capsule shells.

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Section: Optimization Of Pcr-rflpsupporting

confidence: 93%

Section: Primer Characterization Analysismentioning

confidence: 99%

Section: Optimization Of Pcr-rflpmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Rapid Pcr–based Detection Optimization of Porcine Dna in Gelatin Capsule Shell

2018

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“…As at 2013, the global Halal market size was about 580 to 660 billion US Dollars and has great potential to increase in the future year ( Jahangir et al 2016 ). Apart from increasing Muslim population and Halal awareness, Halal market is driven by development of Halal ingredients authentication methods that strengthening consumers’ confident ( Malik et al 2016 ). Gelatin is among the most studied Halal ingredient because of its vast usage in pharmaceutical and food products.…”

Section: Introductionmentioning

confidence: 99%

A Review of Gelatin Source Authentication Methods

Asiyanbi-H

Idris

et al. 2018

TLSR

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Gelatin is a very popular pharmaceutical and food ingredient and the most studied ingredient in Halal researches. Interest in source gelatin authentication is based on religious and cultural beliefs, food fraud prevention and health issues. Seven gelatin authentication methods that have been developed include: nucleic acid based, immunochemical, electrophoretic analysis, spectroscopic, mass-spectrometric, chromatographic-chemometric and chemisorption methods. These methods are time consuming, and require capital intensive equipment with huge running cost. Reliability of gelatin authentication methods is challenged mostly by transformation of gelatin during processing and close similarities among gelatin structures. This review concisely presents findings and challenges in this research area and suggests needs for more researches on development of rapid authentication method and process-transformed gelatins.

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