Commenced in January 2007
Paper Count: 30455
Comparison of Real-Time PCR and FTIR with Chemometrics Technique in Analysing Halal Supplement Capsules
Abstract:Halal authentication and verification in supplement capsules are highly required as the gelatine available in the market can be from halal or non-halal sources. It is an obligation for Muslim to consume and use the halal consumer goods. At present, real-time polymerase chain reaction (RT-PCR) is the most common technique being used for the detection of porcine and bovine DNA in gelatine due to high sensitivity of the technique and higher stability of DNA compared to protein. In this study, twenty samples of supplements capsules from different products with different Halal logos were analyzed for porcine and bovine DNA using RT-PCR. Standard bovine and porcine gelatine from eurofins at a range of concentration from 10-1 to 10-5 ng/µl were used to determine the linearity range, limit of detection and specificity on RT-PCR (SYBR Green method). RT-PCR detected porcine (two samples), bovine (four samples) and mixture of porcine and bovine (six samples). The samples were also tested using FT-IR technique where normalized peak of IR spectra were pre-processed using Savitsky Golay method before Principal Components Analysis (PCA) was performed on the database. Scores plot of PCA shows three clusters of samples; bovine, porcine and mixture (bovine and porcine). The RT-PCR and FT-IR with chemometrics technique were found to give same results for porcine gelatine samples which can be used for Halal authentication.
Digital Object Identifier (DOI): doi.org/10.5281/zenodo.1474717Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 320
 Phillips, Handbook of Hydrocolloid Second Edition. Cambridge: Woodhead publishing limited UK, 2009.
 G. Zhang, T. Liu, Q. Wang, L. Chen, and J. Lei, “Mass Spectrometric Detection of Marker Peptides in Tryptic Digests of Gelatin: A New Method to Differentiate Between Bovine and Porcine Gelatin,” J. Food Hydrocolloids, vol. 23, 2009, pp. 2001-2007.
 G.S Al-Saidi, A. Al-Alawi, M.S. Rahman, and N. Guizani, “Fourier Transform infrared (FTIR) Spectroscopic Study of Extracted Gelatin from Shaari (Lithirinus microdon) Skin:Eeffects of Extraction Conditions”, IFRJ, vol. 19, 2012, pp. 1167-1173.
 N. Azira. I. Amin, and Y.B. Che Man, “Differentiation of Bovine and Porcine Gelatins in Processed Products Via Sodium Dodecyl Sulphate-Polyacrylamide Gel Electrophoresis (SDS-PAGE) and Principal Component Analysis (PCA) Techniques,” Int Food Rsch, vol. 19, 2012, pp. 1175-1180.
 A. Widyaninggar, Triwahyudi, K. Triyana, and Ab. Rohman, “Differentiation Between Porcine and Bovine Gelatin in Commercial Capsule Shells based on Amino Acid Profiles and Principal Component Analysis”, Indonesian J.Pharm, vol. 23, 2012, pp. 96-101.
 BioRad, Real-Time PCR Application Guide, May 2016.
 S. Soares, J.S. Amaral, M.B.P.P Oliveira, and I. Mafra, “A SYBR Green Real-Time PCR Assay to Detect and Quantify Pork Meat in Processed Poultry Meat Products,” Meat Science, vol. 94, 2013, pp. 115–120.
 S. Tanabe, M. Hase, T. Yano, M. Sato, T. Fujimura, and H. Akiyama, “Real-Time Quantitative PCR Detection Method for Pork, Chicken, Beef, Mutton, and Horseflesh in Foods,” Bioscience. Biotechnology. Biochemistry, vol. 71, 2007, pp. 5-3131.
 R. Higuchi, C. Fockler, G. Dollinger, and R. Watson, “Kinetic PCR Analysis: Real-time Monitoring of DNA Amplification Reactions,” J. Biotech, vol. 11, 1993, pp. 1026-1030.
 J.H. Muyonga, C.G.B. Cole, and K.G. Duodu, “Fourier Transform Infrared (FTlR) Spectroscopic Study of Acid Soluble Collagen and Gelatin from Skin and Bones of Young and Adult Nile Perch (L.niIoticus),” J. Food Chemistry, vol. 86, 2004, pp. 325-332.
 S. Krimm, and J. Bandekar, “Vibrational Spectroscopy and Conformation of Peptides, Polypeptides and Proteins”. Advances in Protein Chemistry,” vol. 38, 1986, pp. 181-364.
 G. Fischer, X. Cao, N. Cox and M. Francis, “The FTIR Spectra of Glycine and Glycylglycine Zwitterions Isolated in Alkali Halide Matrices,” J. Chem. Phys, vol. 313, 2005, pp. 39-49.
 P. Lagant, G. Vergoten, M.H, Loucheux-Lefebvre, and G. Fleury, “Raman Spectra and Normal Vibrations of Dipeptides I. Glycylglycine,” J. Bio. Polymers, vol. 22, 1983, p. 1267.
 A.M. Marina, Y.B. Che Man, and A. Ismail, “Use Of The SAW Sensor Electronic Nose for Detecting The Adulteration of Virgin Coconut Oil with RBD Palm Kernel Olein,” J.O.T.A.O.S, vol. 87, 2010, pp. 263-270.
 A. Rohman, and Y.B.Che Man, “Analysis of Pig Derivatives for Halal Authentication Studies,” J. Food Rev Int. Vol. 28, 2012, pp. 97-112.