Post on 11-May-2015
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
©2012 Waters Corporation 2
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)
©2012 Waters Corporation 3
Why is Testing for Mycotoxins Important?
Conformance to regulations
y g y p
Quality, Productivity, Animal Health:Customer Specifications
Brand protection
©2012 Waters Corporation 4
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
©2012 Waters Corporation 5
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
©2012 Waters Corporation 6
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
©2012 Waters Corporation 7
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
©2012 Waters Corporation 8
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)
©2012 Waters Corporation 9
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
©2012 Waters Corporation 10
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
©2012 Waters Corporation 11
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
©2012 Waters Corporation 12
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
©2012 Waters Corporation 13
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!
©2012 Waters Corporation 14
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.
©2012 Waters Corporation 15
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
©2012 Waters Corporation 16
Enzyme Linked Immunosorbent Assay (ELISA)( )
©2012 Waters Corporation 17
LC-MS
©2012 Waters Corporation 18
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
©2012 Waters Corporation 19
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
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
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
©2012 Waters Corporation 22
Now on to Sampling!Now on to Sampling!
©2012 Waters Corporation 23