Feed webinar _VICAM

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

Descriptionp

This webinar will :— explain why it is important to your business to test for mycotoxinsexplain 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 convince you that businesses of all sizes can protect themselves

from mycotoxin risks and meet customer demand for uncontaminated products


What are Mycotoxins?y

Mykes: Greek for fungus/mold

h l d d b ld

Toxicum: Latin for poison/toxin

Mycotoxins are chemicals produced by molds

Molds can grow on plants both before harvest and during storage

ff ld d d ff Different molds can produce different mycotoxins

Different mycotoxins have different toxicity profiles

Regulations for mycotoxins vary (species, country, commodity)


Why is Testing for Mycotoxins Important?

Conformance to regulations

y g y p

Quality, Productivity, Animal Health:Customer Specifications

Brand protection


Most Common Mycotoxins regulated in Human Foods and Feed Worldwide

dMycotoxin Human Feed

Total Aflatoxin 76 49

Aflatoxin B1 61 48

Aflatoxin M1 60 0Aflatoxin M1 60 0

Patulin 48 0

Ochratoxin A 37 36

Deoxynivalenol 37 36

Human: FOA Food and Nutrition Paper #81, 2003

F d K t i / l ti ht 2012

Zearalenone 17 35

Fumonisin 6 29

Feed: www.Knowmycotoxins.com/regulations.htm, 2012


US Aflatoxin Regulations Foodg

C d l ( b) fCommodity Action Level (ppb) Reference

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

US Aflatoxin Regulations for Feedg

Class of Animals Feed Aflatoxin Level

Fi i hi b f ttl C d t d t 300 ppbFinishing beef cattle Corn and peanut products 300 ppb

Beef cattle, swine or poultry Cottonseed meal 300 ppb

Finishing swine over 100 lb. Corn and peanut products 200 ppb

Breeding cattle, breedingswine and mature poultry

Corn and peanut products 100 ppbswine and mature poultry

Immature animals Animal feeds and ingredients, excluding cottonseed meal 20 ppb

Dairy animals, animals not listed above, or unknown use Animal feeds and ingredients 20 ppb


Source: http://www.fda.gov/AnimalVeterinary/Products/AnimalFoodFeeds/Contaminants/ucm050974.htm

Aflatoxin Requirements in the EC

Aflatoxin B 1Maximum content in mg/kg (ppm) relative to a feedingstuff with a moisture content of 12%Aflatoxin B 1 feedingstuff with a moisture content of 12%

All feed materials 0.02

Complete feedingstuffs for cattle, sheep and goats with the exception of:- complete feedingstuffs for dairy animals

0.02

0.005- complete feedingstuffs for calves and lambs 0.01

Complete feedingstuffs for pigs and poultry (except young animals) 0.02

Other complete feedingstuffs 0.01

Complementary feedingstuffs for cattle, sheep and goats (except complementary feedingstuffs for dairy animals, calves and lambs)

0.02

Complementary feedingstuffs for pigs and poultry (except young animals) 0.02(except young animals)

Other complementary feedingstuffs 0.005


Source: www.food.gov.uk/safereating/mycotoxins/animalfeed/

Regulatory Requirements in the ECg y q

Aflatoxin B1:― Commission Directive 2003/100/EC of 31 October 2003Commission 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— 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 (2006/576/EC)( 006/5 6/ C)


Worldwide requirements: Japan

Japan Compound feeds for cattle (except calves, dairy cows), pigs (except piglets), chicken (except young chicken broilers) quails

Aflatoxin B1 20

q p

(except young chicken, broilers) quails

Compound feeds for calves, dairy cows, piglets, young chicken and broilers Aflatoxin B1 10

Compound feeds ZearalenoneDON

10001000

Compound feeds for cows with an age over 3 months DON 4000

Complete feed for beef cow (excluding calves), swine (excluding piglet), poultry (excluding hi k d il)

Aflatoxin B1 20chicks and quail)

Complete feed for calves, dairy cows, piglets, chicks and broiler starters Aflatoxin B1 200

Feed for all livestock Zearalenone 20

S li t f 39 t i th EC K t i / l ti ht

Feed for all livestock (excluding calves <3months) DON 200

Feed for calves <3months DON 4000


Source: list of 39 countries + the EC; www.Knowmycotoxins.com/regulations.htm

Animal Health and Productivity

Toxin Organ Affected Symptoms

y

Aflatoxin Liver Enlarged, Friable

Ochratoxin Kidney Enlarged, Inflammedy g

T2 toxin Mouth, Gizzard Ulcers, Erosion

Zearalenone Uterus Ovary Enlarged InflammedZearalenone Uterus, Ovary Enlarged, Inflammed

Deoxynivalenol Liver Size reduction

Adapted from: http://en.engormix.com/MA-pig-industry/health/articles/target-organs-key-effective-t1241/p0.htm

Fumonisin Lungs, Heart Enlarged


How can I test for Mycotoxins?

Techniques for mycotoxin testing range from sophisticated analytical instruments to test strips similar to a home

y

analytical instruments to test strips similar to a home pregnancy test

Tests can be done in-house or sent to commercial analytical labs


Technologies for Analysis of Mycotoxinsg y yInstrumentation costTraining requiredPrecision and accuracyInformation contentVersatility

HigherLower

LC-MSUPLC-MS

HPLC, GCIAC + FluorometerELISA

Strip tests

Tlc

Immunoassay Instrumental methods u oassay st u e ta et ods


Some sample preparation necessary for all methods

Ground sample Add solvent Vortex, Filter, settle G ou d sa p e Add solventBlend etc.

Filter, settle centrifuge or

other

Most samples will be solids Grind to appropriate fineness Use appropriate solvent for the analyte Use appropriate solvent for the analyte Sample must be representative!


Typical Qualitative (yes/no) Strip Testyp Q (y / ) p

Results in as little as 3 minutesThe presence of two pink lines

– the test line and the control line –indicates a NEGATIVE result (a level of less than 10 ppb).

To check for a POSITIVE result (equal to or greater than 10 ppb),

let the AflaCheck test strip develop for at least 5 minutesbefore reading. If the test line hasn’t appeared

after 5 minutes, the result is positive.


Workflow Quantitative Strip Test (VICAM DON-V) ( )

Wheat, Barley or 5 g wheat + 20 Vortex Let filter 3 , yMaize sample

gmL water 1 minute minutes or less

200 µL filtrate Add 200 µL DON-V

Diluent

Apply 100 µL

to sample well

Read test

at 4 minutes


Enzyme Linked Immunosorbent Assay (ELISA)( )


LC-MS


Technologies for Analysis of Mycotoxinsg y yInstrumentation costTraining requiredPrecision and accuracyInformation contentVersatility

HigherLower

LC-MSUPLC-MS

HPLC, GCIAC + FluorometerELISA

Strip tests

Tlc

Immunoassay Instrumental methods u oassay st u e ta et ods


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?


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 ?


Considerations when choosing a testing strategygy

Testing volume

Turn-around time

Labor - appropriate level of education

Capital investment and consumables

Waste disposal

COA from suppliers


Now on to Sampling!Now on to Sampling!


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