analytical aspects of mycotoxin binders · 2010. 3. 3. · lower analytical values for any...

http://www.jrc.ec.europa.eu/

Joint Research Centre (JRC)

Fusarium-toxin Forum of 1 - 2 February 2010 1

Analytical Aspects of Mycotoxin Binders

Anna Kolossova & Joerg Stroka


Overview

Legal background

Type of Binders (structure)

Binders available to test in the study

Design/outline of study and evaluation

Analytical methods used

Statistical evaluation

Conclusion


Legislation first…


Some background

My first contact with “binders”…

Humans in Ghana

Macaws in Peru

Munn, C.A., 1994. Macaws: winged rainbows. Natl. Geograph. 185:118-140.

TD Phillips is working with Don Brightsmith, Head of Rainforest Expeditions, to determine the reason for geophagy in Macaws in Peru.

http://www.inkanatura.com/macawclaylicks.asp


What has research been focussed on?

Animal health (in vivo)

Metabolic indicators (in vivo)

Cell culture studies (in vitro)

Isotherm studies (analysis of suspensions)

Effects on analysis????


Methods for prevention and decontamination of mycotoxins

Prevention of mycotoxin contaminationPre- and post-harvest control and harvest management

Codex Alimentarius – codes of practice for the prevention and reduction of mycotoxins in cereals, peanuts, apple products, raw materialsThe use of HACCP – to consider in the futureAlthough the prevention of mycotoxin contamination in the field is the main goal of agricultural and food industries, under certain environmental conditions the contamination of various commodities with mycotoxins is unavoidable

Detoxification of mycotoxinsRemoval from contaminated commoditiesInactivation of mycotoxins by physical, chemical or biological methodsNeither the use of chemical decontamination processes, nor the mixing of batches with the aim of decreasing the level of contamination below the maximum tolerable level are legal within the EU

Mycotoxins can not be entirely eliminated


Mycotoxin binders

A mycotoxin binder (adsorbent/denaturant) added to the diet should be able to interact in vivo with mycotoxins and restrict their absorption in the GI tract of animals

Several requirements for a mycotoxin binder

Effective inactivation of mycotoxins of interestReduction of mycotoxin bioavailability/activityAvailability of essential nutrients to an animalNot being detrimental to animal or food productNot being a growth promoter (growth promotion may mask mycotoxins)Demonstrate verifiable positive results (improvement of zootechnicalperformance; recovery of organ status; excretion of mycotoxins via faeces; recovery of the immune status)Technical and economical feasibility


Types of Mycotoxin Binders

AdsorbentsInorganic

Activated charcoalAluminosilicates, clays

HSCAS, bentonite, montmorillonite, zeolite, clinoptilolite…Clay minerals – layered silicates [Si2O5

2-]xyZeolite – silicate made of tetrahedrons of SiO4 and AlO4

Chemically treated silicates

OrganicChemical polymers (cholestyramine, polyvinylpyrrolidone, divinylbenzene-styrene polymers…) High costsGlucan products

Yeast and products from yeast (yeast cell wall)


Types of Mycotoxin Binders

DenaturantMycotoxin-degrading enzymesLife microorganisms producing mycotoxin-degrading enzymes

Eubacterium sp. (BBSH 797)T. mycotoxinivorans

Organic bindersEfficient against a larger range of mycotoxins than inorganic bindersMore adapted to the most frequent cases of multi-contaminated feedsBiodegradable and do not accumulate in the environment after being excreted by animals


Oltipraz a “Mycotoxin Binder”?

http://phm.utoronto.ca/~jeffh/jnppres.ppt


Level of Interest for Testing Mycotoxin Binders


Analytical aspects

Cascade of modern analytical Methods:

– Extraction

– Clean-up / Dilution

– Separation

– Detection


Methods used for testing

Official Methods (or those in standardization process):

• Aflatoxin B1 EN ISO 17375:2006

• Deoxynivalenol prEN 15791:2009

• Zearalenone prEN 15792:2009

• Ochratoxin A WD N954 of TC327/WG1

• Fumonisins WD N953 of TC327/WG1

• T2 and HT2 toxins Breidbach et.al. under consideration as a prospective CEN standard.


Repeatability Data of the Test Methods

4.1OTA

3.2FUM B1 + B2

9.8T2 & HT2 toxins

7.7ZEA

6.9AFB1

10.6DON

Average RSDr (%)Mycotoxin


Tested Binders in this Study

2.55.1Fibres (lignocellulose)20

0.59.8HSCAS19

0.59.8HSCAS18

0.58.8HSCAS17

0.410.6Montmorillonite16

0.410.4Montmorillonite15

16.7Fibres14

18.9Clay13

0.59.1Clay12

0.256.9Mixture organic + mineral component11

0.258.6Mixture organic + mineral component10

0.25.0Yeast cell wall9

0.256.2Mixture organic + mineral8




0.254.6Clay4

0.18.4Clay3

0.17.9Clay + organic acid2


Amount added [%]pH valueType of main product(s)Code


Some Statistics

Data analysisStudent’s t-test (independent two-sample t-test with unequal sample sizes and equal variance). Differences were assumed to be significant at p < 0.05.

Critical t-values (p = 0.05) were compared with those obtained by the Student’s t-test according to the following formula:

21

11

21

nnS

XXt+×

−=

Statistical Advisor: A. Breidbach


Binder

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20

Ret

rieva

l of a

flato

xin

B1,

%

0

20

40

60

80

100

120

Aflatoxin B1


DON

Binder

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20

Ret

rieva

l of d

eoxy

niva

leno

l, %

0

20

40

60

80

100

120


ZON (combined binders)

Binder group

A B C D E

Ret

rieva

l of z

eara

leno

ne, %

0

20

40

60

80

100


OTA (combined binders)

Binder group

A B C D E

Ret

rieva

l of o

chra

toxi

n A

, %

0

20

40

60

80

100


Fumonisins (combined binders)

Binder group

A B C D E

Ret

rieva

l of f

umon

isin

s B

1 & B

2, %

0

20

40

60

80

100


T-2 & HT-2 toxins

Binder group

A B C D E

Ret

rieva

l of T

-2 &

HT-

2 to

xins

, %

0

20

40

60

80

100

120


Recent Findings (FAC-A) on AfB1


Recent Findings (FAC-A) on DON


AfB1 to AfM1 transformation in the presence of binders


Conclusion

The addition of the tested binders (at the level indicated by the manufacturer) did not result in lower analytical values for any mycotoxincurrently regulated in feed.

The combined effect of processing such as wet heating, pH-change, etc. in the presence of mycotoxin binders was not investigated, but might be a subject of interest for other studies.


Collaborators

Ouissam Abbas and Vincent BaetenIn collaboration with

Alexandro Rodriguez

Walloon Agricultural Research CentreQuality of Agricultural Products Department

Chaussée de Namur, 24B – 5030 GEMBLOUX

Tél :++ 32 (0) 81/62.03.50Fax : ++ 32 (0) 81/62.03.88

[email protected]://cra.wallonie.be


IRMM staff behind the scene

Massimo AmbrosioKaty KroegerKatrien Bouten


Open Call for Collaborators

IRMM is looking for collaborators for projects on:characterisation studies homogeneity and stability studies, and feasibility studies

As well as for scientific staff (e.g. post docs)

www.irmm.jrc.be/html/calls/call_for_collaborators/index.htmwww.irmm.jrc.be/html/job_opportunities/index.htm

analytical aspects of mycotoxin binders · 2010. 3. 3. · lower analytical values for any...

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