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Detection of Non-Halal Ingredients_2014

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This presentation was given by Associate Professor Shuahaimi Mustafa, Universiti Putri Malaysia, at the Vita Foods Conference in Hong Kong, 2-3 September 2014

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Detection of Non-Halal Ingredients_2014

  1. 1. DETECTION OF NON-HALAL INGREDIENTS Shuhaimi Mustafa Halal Products Research Institute (IPPH), UPM 3-4 SEPTEMBER 2014 AsiaWorld-Expo, Hong Kong
  2. 2. Contents Background Current issues in halal products Halal analysis and authentication Conclusion
  3. 3. حلالاً طيبا Halal – permissible by Shariah Law. Haram – prohibited by Shariah Law. Thoyyib – Good and wholesome (quality, safe, nutritious, pure).
  4. 4. Muslims cannot consume the following: •pork or pork by products, •animals that were dead prior to slaughtering, •animals not slaughtered properly or not slaughtered in the name of Allah, •blood and blood by products, •alcohol, •carnivorous animals, •birds of prey, •land animals without external ears
  5. 5. The needs for analysis Religious beliefs Allergic Fair-trade Laidan (2011) stated that a particular Halal-certified product cannot be genuinely guaranteed halal without being tested in halal laboratory. The following examples justify the need for analysis: From 902 meat products tested; 15.9% cases for raw product containing undeclared animal species and 22.9% cases for cooked products containing undeclared animal species. ~50% animal feeds contain undeclared animal species.
  6. 6. Meat-forgery scandals
  7. 7. Status of some food additives/ingredients Food additive/ingredients Description Status Riboflavin Color Haram if from pork liver/kidney Lycopene Color Doubtful, may be mixed with gelatin Butylated hydroxyanisole (BHA) Antioxidant Doubtful, if animal fat is used as carrier Glycerol Sugar alcohols Doubtful, if it is from animal Mono-and diglycerides of fatty acids Emulsifiers Doubtful, if it is from animal
  8. 8. Challenges in halal analyses No pressure/requirement by relevant authorities. Industry wants faster halal certification process Complex analytical techniques. Lack of competent analysts. Undeclared ingredients and processing aids. Lack of biomarkers: - oil/fat-based - protein-based - DNA-based - metabolites-based
  9. 9. Technique Target gene/sequence Reference Conventional PCR 12S rRNA Dalmasso et al. (2004) Taqman Real-time PCR Genomic Cai et al. (2012) Molecular Beacon real time PCR Mitochondrial cytochrome b Mohd Yusop et al. (2012) Commercial Real-time PCR Kit Unknown Demirhan et al. (2012) Technique Sample Reference FTIR Shortening Syahariza et al. (2005) FTIR Gelatin Hashim et al. (2010) FTIR + PLS Meatball Rohman et al. (2011) FTIR + chemometrics Ham sausages Xu et al.(2012) Technique Sample(s) Reference DSC Cooking oils + lard Mansor et al. (2012) Technique Sample(s) Reference MALDI-TOF-MS, LC-MS Porcine gelatine Zhang et al. (2009) HS-SPME-GC-MS Alcoholic beverage Garcia-Martin et al. (2010) GCxGC-TOF-MS Lard Indrasti et al. (2010) GC-MS Pork Nurjuliana et al. (2011) Some methods developed specifically for halal analysis
  10. 10. Halal Products Analyses MS ISO/IEC 17025:2005 Accredited Scope of Accreditation Material/Products tested Type of test Method Collagen, gelatin capsule Amino acid profiling HPLC DNA (meat and meat- based products) Porcine specific DNA marker HaFYSTM
  11. 11. HaFYSTM: 1 step, 1 button, ~ 1 hour Collect Sample Place in Test Module Insert into Analyzer On one touch Yes/No Results
  12. 12. Red line: Internal control; blue line: beef sample (Negative Result)
  13. 13. Red line: Internal control; blue line: Cat food containing pork (Positive Result)
  14. 14. Red line: Internal control; blue line: no- template control
  15. 15. Animal feeds
  16. 16. Amino acid analysis of gelatin using HPLC and Chemometric approaches Stages in analysis Explanation Acid hydrolysis hydrochloric acid is the most widely used agent for hydrolyzing proteins. the hydrolysis process occurred in 110OC within 24hrs Pre-column derivatization (Using AccQ tag Fluor reagent ) using 6-aminoquinolyl-N- hydroxysuccinimidyl carbamate derivatives Quantification separations by HPLC-FD All amino acid derivative separations required a gradient chromatography due to the wide hydrophobicity range of the amino acid side chains use fluorescence detection with excitation at 248nm and emission at 395nm.
  17. 17. PCA VIEW – SCORES PLOT
  18. 18. Projection of unknown samples Gelatin capsules – Nitta (Japan), Gelita (india), Sterling (brazil) & Rousoullet (china).
  19. 19. Porcine samples Porcine (Merck) Porcine (Fluka) Porcine skin gelatin Porcine(Dr.Oetker) Porcine (Tessenderlon)
  20. 20. Alcohol in products (GC/MS method)
  21. 21. y = 0.0001x - 0.0019 R² = 0.9982 0.0000 0.2000 0.4000 0.6000 0.8000 1.0000 1.2000 1.4000 1.6000 0 1000 2000 3000 4000 5000 6000 7000 8000 9000 10000 11000 ETOH/ACN PEAK AREA ETHANOL CONCENTRATION (PPM) HIGH ETHANOL CALIBRATION CURVE (1000 - 10 000 PPM)
  22. 22. Calculation of Ethanol Content By taking the equation y = 0.0001x - 0.0019, the amount of ethanol present in sample (ex : Sushi dipping sauce) can be calculated. Y = Peak area of Ethanol / Peak area of Acetonitrile x = Ethanol Concentration (ppm) When y = 0.6204, x = 0.6204 + 0.0019/0.0001 = 6222.916 ppm or 0.6223% Multiply by 10 (1 gm of original sample was diluted with deionized water up to 10 ml ) So, original sample contains 6.223% w/v ethanol (considered as HARAM).
  23. 23. Alcohol content in beverages Beers (3-10%) Wines (8-14%) Fortified wines (16-22%) Fruit juices (<0.1%)
  24. 24. Conclusion Halal should take into account beyond just trust, social, spiritual, environmental, and sustainability issues through document audit-based and site visit approaches as practiced by almost all halal certification bodies, but must also be verified through laboratory analysis.

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