Feed webinar _VICAM


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Feed webinar _VICAM

  1. 1. Webinar: Best practices for sampling and testing of mycotoxins  Discover 5 reasons why you should be testing for mycotoxins and avoid unnecessary risks.  Compare the merits of different testing methods.  Learn the most recommended sampling practices.©2012 Waters Corporation 1
  2. 2. Description p  This webinar will : — explain why it is important to your business to test for mycotoxins — describe the advantages and disadvantages of different testing methods and what to look for when choosing a method — convince you that businesses of all sizes can protect themselves from mycotoxin risks and meet customer demand for uncontaminated products©2012 Waters Corporation 2
  3. 3. What are Mycotoxins? y Mykes: Greek for fungus/mold Toxicum: Latin for poison/toxin  Mycotoxins are chemicals produced b molds h l d d by ld  Molds can grow on plants both before harvest and during storage  Different molds can produce d ff ff ld d different mycotoxins  Different mycotoxins have different toxicity profiles  Regulations for mycotoxins vary (species, country, commodity)©2012 Waters Corporation 3
  4. 4. Why is Testing for Mycotoxins Important? y g y p  Conformance to regulations  Quality, Productivity, Animal Health:Customer Specifications  Brand protection©2012 Waters Corporation 4
  5. 5. Most Common Mycotoxins regulated in Human Foods and Feed Worldwide Mycotoxin Human Feed d Total Aflatoxin 76 49 Aflatoxin B1 61 48 Aflatoxin M1 60 0 Patulin 48 0 Ochratoxin A 37 36 Deoxynivalenol 37 36 Zearalenone 17 35 Fumonisin 6 29 Human: FOA Food and Nutrition Paper #81, 2003 Feed: F d www.Knowmycotoxins.com/regulations.htm, 2012 K t i / l ti ht©2012 Waters Corporation 5
  6. 6. US Aflatoxin Regulations Food g Commodity C d Action Level ( b) l (ppb) Reference f Foods 20 CPG 555.400 Milk 0.5 (aflatoxin M1) CPG 527.400 Peanuts and Peanut products 20 CPG 570.375 Pistachio nuts 20 CPG 570.500 http://www.fda.gov/Food/GuidanceComplianceRegulatoryInformation/Guida nceDocuments/ChemicalContaminantsandPesticides/ucm077969.htm#afla©2012 Waters Corporation 6
  7. 7. US Aflatoxin Regulations for Feed g Class of Animals Feed Aflatoxin Level 300 ppb Finishing b f cattle Fi i hi beef ttl Corn and peanut products C d t d t Beef cattle, swine or poultry Cottonseed meal 300 ppb Finishing swine over 100 lb. Corn and peanut products 200 ppb Breeding cattle, breeding Corn and peanut products 100 ppb swine and mature poultry Animal feeds and ingredients, Immature animals 20 ppb excluding cottonseed meal Dairy animals, animals not listed Animal feeds and ingredients 20 ppb above, or unknown use Source: http://www.fda.gov/AnimalVeterinary/Products/AnimalFoodFeeds/Contaminants/ucm050974.htm©2012 Waters Corporation 7
  8. 8. Aflatoxin Requirements in the EC Maximum content in mg/kg (ppm) relative to a Aflatoxin B 1 feedingstuff with a moisture content of 12% All feed materials 0.02 Complete feedingstuffs for cattle, sheep and goats 0.02 with the exception of: - complete feedingstuffs for dairy animals 0.005 - complete feedingstuffs for calves and lambs 0.01 Complete feedingstuffs for pigs and poultry (except 0.02 young animals) Other complete feedingstuffs 0.01 Complementary feedingstuffs for cattle, sheep and goats (except complementary feedingstuffs for dairy 0.02 animals, calves and lambs) Complementary feedingstuffs for pigs and poultry 0.02 (except young animals) Other complementary feedingstuffs 0.005 Source: www.food.gov.uk/safereating/mycotoxins/animalfeed/©2012 Waters Corporation 8
  9. 9. Regulatory Requirements in the EC g y q  Aflatoxin B1: ― Commission Directive 2003/100/EC of 31 October 2003 amending Directive (EC) No 2002/32/EC of the European Parliament and of the Council on undesirable substances in animal feed  DON, zearalenone, ochratoxin, T-2, HT-2 and fumonisins — Commission Recommendation of 17 August 2006 on the presence of dexynivalenol, zearalenone, ochratoxin A, T-2 and HT-2 and fumonisins in products intended for animal feeding ( 006/5 6/ C) (2006/576/EC)©2012 Waters Corporation 9
  10. 10. Worldwide requirements: Japan q p Japan Compound feeds for cattle (except calves, dairy cows), pigs (except piglets), chicken Aflatoxin B1 20 (except young chicken broilers) quails chicken, Compound feeds for calves, dairy cows, piglets, Aflatoxin B1 10 young chicken and broilers Zearalenone 1000 Compound feeds DON 1000 Compound feeds for cows with an age over 3 DON 4000 months Complete feed for beef cow (excluding calves), swine (excluding piglet), poultry (excluding Aflatoxin B1 20 chicks and quail) hi k d il) Complete feed for calves, dairy cows, piglets, Aflatoxin B1 200 chicks and broiler starters Feed for all livestock Zearalenone 20 Feed for all livestock (excluding calves DON 200 <3months) Feed for calves <3months DON 4000 Source: list of 39 countries + the EC; www.Knowmycotoxins.com/regulations.htm S li t f t i th EC K t i / l ti ht©2012 Waters Corporation 10
  11. 11. Animal Health and Productivity y Toxin Organ Affected Symptoms Aflatoxin Liver Enlarged, Friable Ochratoxin Kidney y Enlarged, Inflammed g T2 toxin Mouth, Gizzard Ulcers, Erosion Zearalenone Uterus, Uterus Ovary Enlarged, Enlarged Inflammed Deoxynivalenol Liver Size reduction Fumonisin Lungs, Heart Enlarged Adapted from: http://en.engormix.com/MA-pig-industry/health/articles/target-organs-key-effective-t1241/p0.htm©2012 Waters Corporation 11
  12. 12. How can I test for Mycotoxins? y  Techniques for mycotoxin testing range from sophisticated analytical instruments to test strips similar to a home pregnancy test  Tests can be done in-house or sent to commercial analytical labs©2012 Waters Corporation 12
  13. 13. Technologies for Analysis of Mycotoxins g y y Instrumentation cost Training required Precision and accuracy Information content VersatilityLower Higher Strip tests IAC + Fluorometer HPLC, GC LC-MS ELISA UPLC-MS Tlc Immunoassay u oassay Instrumental methods st u e ta et ods©2012 Waters Corporation 13
  14. 14. Some sample preparation necessary for all methods G ou d sample Ground sa p e Add solvent Vortex, Filter, settle Blend etc. centrifuge or other  Most samples will be solids  Grind to appropriate fineness  Use appropriate solvent for the analyte  Sample must be representative!©2012 Waters Corporation 14
  15. 15. Typical Qualitative (yes/no) Strip Test yp Q (y / ) p Results in as little as 3 minutes To check for a POSITIVE result The presence of two pink lines (equal to or greater than 10 ppb), – the test line and the control line – let the AflaCheck test strip develop for at least 5 minutes indicates a NEGATIVE result before reading. If the test line hasn’t appeared (a level of less than 10 ppb). after 5 minutes, the result is positive.©2012 Waters Corporation 15
  16. 16. Workflow Quantitative Strip Test ( (VICAM DON-V)) Wheat, Barley or , y 5 g wheat + 20 Vortex Let filter 3 Maize sample mL water minutes or less 1 minute 200 µL filtrate Add 200 µL DON-V Apply 100 µL Read test Diluent to sample well at 4 minutes©2012 Waters Corporation 16
  17. 17. Enzyme Linked Immunosorbent Assay ( (ELISA) )©2012 Waters Corporation 17
  18. 18. LC-MS©2012 Waters Corporation 18
  19. 19. Technologies for Analysis of Mycotoxins g y y Instrumentation cost Training required Precision and accuracy Information content VersatilityLower Higher Strip tests IAC + Fluorometer HPLC, GC LC-MS ELISA UPLC-MS Tlc Immunoassay u oassay Instrumental methods st u e ta et ods©2012 Waters Corporation 19
  20. 20. How do I know my testing is effective? y g  Do you have a quality and HACCP system in place?  Do you take part in proficiency programs such as AOAC, FAPAS, and FGIS?  Is your in-house method validated and approved?  Do you run check samples?  Is your sampling representative?©2012 Waters Corporation 20
  21. 21. How do I know my testing provider is effective?  Do they have a quality system in place?  Are they accredited by state, national, and international regulatory agencies or industry trade organization?  Do they use validated, approved methods (AOAC, GIPSA)?  Do they take part in proficiency programs such as AOAC, FAPAS, and FGIS ?©2012 Waters Corporation 21
  22. 22. Considerations when choosing a testing strategy gy  Testing volume  Turn-around time  Labor - appropriate level of education  Capital investment and consumables  Waste disposal  COA from suppliers©2012 Waters Corporation 22
  23. 23. Now on to Sampling!©2012 Waters Corporation 23