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

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


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)


Why is Testing for Mycotoxins Important?
y g y p

 Conformance to regulations

 Quality, Productivity, Animal Health:Customer Specifications

 Brand protection


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


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


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


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/


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)


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


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


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


Technologies for Analysis of Mycotoxins
g y y
Instrumentation cost
Training required
Precision and accuracy
Information content
Versatility

Lower Higher

Strip tests IAC + Fluorometer HPLC, GC LC-MS
ELISA UPLC-MS
Tlc

Immunoassay
u oassay Instrumental methods
st u e ta et ods


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!


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.


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


Enzyme Linked Immunosorbent Assay
(
(ELISA)
)


LC-MS


Technologies for Analysis of Mycotoxins
g y y
Instrumentation cost
Training required
Precision and accuracy
Information content
Versatility

Lower Higher

Strip tests IAC + Fluorometer HPLC, GC LC-MS
ELISA UPLC-MS
Tlc

Immunoassay
u oassay Instrumental methods
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
strategy
gy

 Testing volume

 Turn-around time

 Labor - appropriate level of education

 Capital investment and consumables

 Waste disposal

 COA from suppliers


Now on to Sampling!


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