REDUCTION,

DECONTAMINATION AND
DETOXIFICATION OF
Sardjono
Faculty of Agricultural Tecnology
!adja Mada Uni"er#ity
DETOXIFICATION OF
M$COTOXINS IN FOOD
Introduction
Occurrence of mycotoxins in foods
The role of food processing on reduction,
CONTENT %
The role of food processing on reduction,
decontamination and detoxification of
mycotoxins
Detoxification by indigenous strain
Conclusion
INDONESIA
TRO&ICA' COUNTR$
(o(ulation % ))* +illion
Self #ufficiency in te
(roduction of ,a#ic food
agricultural cro(# -cereal#,
oter car,oydrate #ource,
,ean#.
/ig u+idity - R/ 0 123.
4ar+ te+(erature -)5 67)8C.
Ideal for fungal growth
I+(ro(erly 9 (oor andling and
#torage condition
MYCOTOXICOSES
Mycotoxigenic fungi and
occurrence of mycotoxins
STORA!E
PRODUCTION
RA4 MATERIA'S FOR
/AR:EST AND &OST
/AR:EST /AND'IN!
&ROCESSED FOODS
PROCESSING
RA4 MATERIA'S FOR
FOOD INDUSTR$
M$COTOXINS
CONTAMINATION IN
FOOD C/AIN
STORA!E
CONSUM&TION
Dry and heat
resistant fungi?
MYCOTOXIN MYCOTOXIN COMODITY COMODITY !"OD#C$" !"OD#C$" %$&'T% "I( %$&'T% "I(
&flatoxins &flatoxins
)*+, *,, -+, -,. )*+, *,, -+, -,.
!eanut, corn, grain and !eanut, corn, grain and
their product their product
Aspergillus Aspergillus flavus flavus
Aspergillus Aspergillus parasiticus parasiticus
Carsinogenic, Carsinogenic,
embryotoxic embryotoxic
&flatoxin M+ &flatoxin M+ Mil/ Mil/ &flatoxins &flatoxins0 0 Contaminted Contaminted
feed feed
Carsinogenic Carsinogenic
1umonisins 1umonisins
) *+,*, . ) *+,*, .
Corn, 2heat and other Corn, 2heat and other
grain grain
Fusarium Fusarium moniliforme moniliforme Carsinogenic Carsinogenic
&ccut &ccut
T/E FI:E FUN!A' M$COTOXINS IN A!RICU'TURA' &RODUCT
3earalenone 3earalenone Corn, 2heat, barley Corn, 2heat, barley
Fusarium Fusarium graminearum graminearum
F. F. Culmorum Culmorum
F. F. crookwellense crookwellense
Carsinogenic Carsinogenic
"eproduction "eproduction
system system
Deoxyni4alenol Deoxyni4alenol
Ni4alenol Ni4alenol Corn, 2heat, barley Corn, 2heat, barley
Fusarium Fusarium graminearum graminearum
F. F. Culmorum Culmorum
F. F. crookwellense crookwellense
Carsinogenic Carsinogenic
"eproduction "eproduction
system system
Ochratoxin & Ochratoxin &
)OT&. )OT&. Coffee, cocoa, 2heat Coffee, cocoa, 2heat
Aspergillus Aspergillus ochraceus ochraceus
Penicillium Penicillium vericosum vericosum carsinogenic carsinogenic
Natural co0occurrence of aflatoxins and
1usariummycotoxins in corn
in corn from South East Asia (Yamashita, et al (1995))
Mean (range) concentration (ng/g) in positive sample
FMB
1
FMB
2
AFB
1
AFB
2


Yamashita, A., Yoshizawa T., Aiura Y., Sanchez P.C., Dizon E.I., Arim R.H.
and Sardjono. 1995. Fusarium mycotoxins (Fumonisins, Nivalenol and
Zearalenone ) and aflatoxins in corn from South East Asia.

Philippines

Thailand

Indonesia


419
(57-1820)
1580
(63-8,800)
843
(226-1780)

286
(58-1210)
251
(50-1400)
442
(231-556)

49
(1-430)
63
(1-606)
352
(1-3300)

14
(1-78)
14
(1-73)
90
(1-680)

Abbrevation : FMB
1
and FMB
1
, Fumonisin B
1
and B
2
respectively; AFB
1
and AFB
2
,
Aflatoxin B
1
and B
2
, respectively
Central Java*)
Mycotoxins
Concentration (ng/g)
Average
Mycotoxins
Concentration (ng/g)
Average

AFB
1

AFB
2

AFG
1


102
20
8

FMB
1

FMB
2

FMB
3


788
182
108
Norhayati Ali, Sardjono, Yamashita, A., and Yoshizawa, T., 1998. Natural Co-
occurence of Aflatoxins and Fusarium Mycotoxins (fumonisins, deoxynevalenol,
nivalenol dan zearalenone) in Corn From Indonesia.
AFG
1
8 FMB
3

DON
NIV
ZEA
108
27
109
12

I+(ortant i##ue tat corn fro+ tro(ical country in te #a+e gro;ing condition#, can
,e #i+ultaneou#ly conta+inated ;it aflato<in#, fu+oni#in#, ni"alenol,
deo<yni"alenol and =earalenone
'e"el of Fu+oni#in# conta+ination in +ai=e6,a#ed
food# and feed fro+ $ogya>arta , Indone#ia
-Nuryono et al?, )**@.A
Sa+(le category
Industrially0produced
foods
Sa+(le nu+,er
&naly5ed positi4e
,6 +6
Fu+oni#in# -((,.
"ange &4erage
,,780+967: ;97+
foods
mall industry0products
Mai5e flour
Mai5e for foods
1ormulated feed
+< +:
6 0
= ;
>9 ,=
+,7=0,>67+ ;676
0 0
:8790,6<+ +,<;
+97<0,,;< =:8
A Mycoto<in Re#earc )*-)**@.%)6B
'e"el of Cearalenone conta+ination in +ai=e6,a#ed
food# and feed fro+ $ogya>arta , Indone#ia
-Nuryono et al?, )**5.A
Sa+(le category Cearalenone -((,.
"ange &4erage
Sa+(le nu+,er
&naly5ed positi4e
Industrially0produced foods
mall industry0products
Mai5e for foods
1ormulated feed
;7;0+87: =7+
,+7,0;,: +==
:7+0:7> :7,
:7>0;,: ,;7;
,+ 6
+< ;
+> ,
,+ +8
A D? of Food Control EF-)**5.%F561E
/UMAN OR!AN TAR!ET
F
O
O
D

P
R
O
C
E
S
S
I
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G
CONTAMINATED
RA4 MATERIA'S
SAFE &RODUCTS
DECONTAMINATION
DETOXIFICATION
/I!/ 'OADG
F
O
P
R
SORTATION
&/$SICA' H
C/EMICA'
&RETREATMENT
NOT FEASII'E FOR 'AR!E SCA'E
SAFE J
&RODUCTION COSTJ
Decontamination should ?
Complete inacti4e, destroy, remo4e the toxins, reduce to
acceptable le4els
Not produce toxic residues in food
!reser4e the nutriti4e 4alue
Not alter acceptability or technological properties Not alter acceptability or technological properties
*e integrated if possible, into regular food0processing and
preparation step
*e cost effecti4e
Not destroy or damage the e@uipment
FOOD PROCESSING
BIOLOGICAL PHYSICAL
CHEMICAL
REDUCTION, DECONTAMINATION,
DETOXIFICATION ?
IIO'O!ICA'
IINDIN!
deconta+ination
IIO'O!ICA'
MET/OD
ENC$MATIC
deto<ification
Organi#+# Action 9 +ecani#+ Refference
A fe; #train# of 'AI to ,ind AFIE in food +odel El6Na=a+i et al?, EBB2
&ieride# et al?, )***
/a#>ard et al?, )**E
non co"alently ,inding AFIE Cang and Ota, EBB7
El6na=a+i et al?, EBB2
Tur,ic et al?,)**)
IINDIN!
Tur,ic et al?,)**)
Lactobacillus ramnosus ,ind to "ia,le and non "ia,le cell El6 Na=a+i et al?, EBB2
a##ociation to ydro(o,ic (oc>et /a#>ard et al?, )***
Staphylococcus aureus to ,ind AFIE Oatley et al?, )***
and Escherichia coli
Lactobacillus and to ,ind AFIE El6Na=a+i et al?, )***
Propionibacterium
L. rhamnosus GG and to ,ind Cearalenone and it# dere6 El6Na=a+i et al?, )**)
L rhamnisus LC70 "ate#
!spergillus ory"ae##$% to ,ind AFIE -reduce in S'F. Sardjono et al?,)**@
&eurospora spp to ,ind AFIE -reduce in S'F. Sigit Setya,udi et al?, )**5
Aflato<in#
deto<ification
OR!ANISMS M$COTOXINS MEDIUM REFFERENCES
'la(obacterium auranotiacum Aflato<in S'F Ciegler et al?, EBFF
Aflato<in ME +il> 'illeoj et al?, EB1E
Aflato<in (eanut +il> /ao and Irac>ett, EBB5
Coryenebacterium rubrum Aflato<in Mann and Re+, EB1F
Candida lipolytica Aflato<in Mann and Re+, EB1F Candida lipolytica Aflato<in Mann and Re+, EB1F
!spergillus niger Aflato<in Mann and Re+, EB1F
)richoderma (iridae Aflato<in Mann and Re+, EB1F
*ucor ambigous Aflato<in Mann and Re+, EB1F
!spergillus niger Aflato<in Faraj et al?, EBB7
+hi"opus sp Aflato<in Iol and S+it, EB2B
Nout, EB2B
&eurospora sp Aflato<in Nout, EB2B
Aflato<in IE S'F Sigit?, FMC et al?, )**5
!spergillus ory"ae ##$% Aflato<in IE S'F Sardjono et al?, )**@
Lactic acid bacteria ,moromi- Aflato<in IE S'F Sardjono et al?, )**5
RESEARCH ON ISOA!ION OF
INDIGENEO"S PO!EN!IA
S!RAINS
Aspergillus oryzae ((*6
E<tracellular en=y+e#
DE!RADATION
DETOXIFICATION DETOXIFICATION
Reduction
Degradation of lactone
O(ening difuran ring
KODI
- Sardjono et al.. )**@.
TOXICIT$ TEST
T/E !RO4T/ OF Iacillu# +egateriu+IN ENC$MATIC
DE!RADATION &RODUCT - Sardjono, et al?, )**@.
A? Nor+al gro;t? I !ro;t on degradation (roduct?
C !ro;t on reaction (roduct for+ eated en=y+e# and AFIE
D$TOXI1IC&TION *Y &'T TO'$"&NT '&CTIC &CID *&CT$"I& IO'&T$D
1"OM MO"OMI ) ardAono, et al7, ,99;.
Lactobacillus #(SS
MOROMI
Lactobacillus #(I!
Lactobacillus #(&S
7 #train# of yea#t
MOROMI
&1*+
DE!RADATION
&RODUCT ,y
Lactobacillus #(I!
MIC"OO"-&NIM MYCOTOXIN M$C%&NIM "$1$""$NC$
Aspergillus niger DONB T0, &cetylation, deacetylation $l har/a2y and &bbas, +==+
Mucor muceda DONB T0, &cetylation, deacetylation $l har/a2y and &bbas, +==+
Agrobacterium rhizobium DON Oxidi5ed to >0/eto060DON hima et al7, +==<
Fusarium nivale DON acetylation of hydroxyl Yoshi5a2a C Moroo/a, +=<;
Fusarium graminearum T0, &cetylation Yoshi5a2a et al7, +=89
'usariumto<in#
deto<ification
Fusarium graminearum T0, &cetylation Yoshi5a2a et al7, +=89
Colonetria nivalis T0, &cetylation Yoshi5a2a et al7, +=89
"corn enzymes" DON reduction Miller and Young, +=8;
Arabidopsis thalia DON %ydroxyl group of DON !oppenberger et al7, ,99>
Fusarium moniliforme 1M*+ &lberts et al7, +==9
!ophiala spinifera 1M*+ %ydroly5ed ester bound Du4ic/, +==6
"hinoclodiella atrovirensa 1M*+ %ydroly5ed ester bound Du4ic/, +==6
!ophilia spinifera 1M*+ Oxidati4e deaminasi *lac/2ell et al7, +===
#hamnidium elegans 3$& 3ea060O0D0glucoside (amimura, +=8:
Mucor baenieri 3$& 3ea060O0D0glucoside $l har/a2y and &bul0%aAA,+=8<
"hizopus spp 3$& 3ea060O0D0glucoside (amimura, +=8:
MICROOR!ANISMS M$COTOXIN MEC/ANISM REFFERENCE
Actinobacter calcoaceticus OT& Degradation *ata and 'as5tity, +===
Aspergillus fumigatus OT& Detoxification Earga et al7, ,999
OTA and &atulin
Aspergillus fumigatus OT& Detoxification Earga et al7, ,999
Aspergillus niger OT& Degradation Earga et al7, ,999
Aspergillus from grape OT& Degradation &brunhosa et al7, ,99,
Paecilomyces !atulin Degradation &nderson et al7, +=<=
yeast from alcoholic fermentation !atulin "eduction %ar2ig et al, +=<=
tinson et al7, +=<8
yeast !atulin Con4ert to acladiol $ C 3 Moss and 'ong, ,99,
$actobacillus plantarum !atulin "educed by intra7 $n5ym "7hapira )unpublished.
T"$&TM$NT M&T$"I&' MYCOTOXIN $11$CT "$11$"$NC$
)**LC, E) +in cofee ,ean aflato<in 1B3 reduction Scott, EB2@
)**LC, E5 +in B@3 reduction
E**LC, ) cotton#eed +eal 7*3 MC aflato<in 5*3 reduction Mann, et al?, EBF1
E**LC, ) cotton#eed +eal F,F*3 MC 5*3 reduction
E15LC corn, dry condition citrinin deco+(o#ed Kita,ata>e et al, EBBE
EF*LC corn, +oi#t citrinin de#troyed
Roa#ting -EB*6))1LC cofee ,ean OTA FB6BF3 reduction "an der Stegen, )**E
for 56)* +in.
%$&TIN-
for 56)* +in.
)**6)5*LC dry or ;et grain OTA co+(lete de#truc6 Ioudra et al?,EBB5
tion
E5*LC, @@ corn Cearalenone not de#tryed Iennet et al?,EB2*
E5*LC6 )**LC corn Fu+oni#in IE 216E**3 de#truct?
)E2LC, E5 +in corn+eal ;it5 Mg9g Fu+oni#in IE co+(lete lo## Ca#telo et al?,EBB2
Ia>ing DON 5*3 lo## Tana>a et al?,EB2F
E<tru#ion coo>ing flour DON B53 reduction Ca==aniga et al?,)**E
E<tru#ion coo>ing flour aflato<in E*6)53 reduction Elia#6Oro=>o, )**E
E<tru#ion -*?73
li+eN E?53 /)O) tortilla# aflato<in co+(lete de#troy
E**LC, O acti"ated a((le juice (atulin al+o#t all Kada>al et al?,)**)
carcoal Kada>al and Na#, )**7
CEREA'S
&ROCESSIN!
C/EMICA'
&/$SICA'
IIO'O!ICA'
C'EANIN!,
DE/USKIN!
&O'IS/IN!
DR$ MI''IN!
4ET MI''IN!
IAKIN!
ROASTIN!
FR$IN!
N
S

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A
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A
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E
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M
A
T
E
R
I
A
L
S
IIO'O!ICA'
FR$IN!
COOKIN!
EXTRUSION
MA'TIN! H
IRE4IN!
&RODUCTSJJ
M
Y
C
O
T
O
X
I
N
R
A
W

M
C'EANIN!, DE/USKIN!, &O'IS/IN!
-enerate little heat no significant thermal brea/do2n
Molds and mycotoxins are concentrated in dust, bro/en
grains or seed coat
urface cleaning, 2ashing and drying, steaming reduction of
&1*+ 2ere >;F, 6<08;F and =90=>F )(utit and Mer/o, &1*+ 2ere >;F, 6<08;F and =90=>F )(utit and Mer/o,
+==+.
!olishing remo4e NIE, DON and 3$&, all 2ere found in bran
fraction, except NIE also found in pearled barley
In mai5e commercial mill &1 and 1M* reduced by >9069F, no
reduction on 3$&
DR$ MI''IN!
Mo#t +ycoto<in# tend to cencentrated in te ,ran fraction
and outer layer of grain (roduce fraction ;it lo;er
concentartion of +ycoto<in#
AFIE % (roduct# -corn+eal or four conta+inated only 16E*3 f
fro+ original AFIE
OTA % lo;er in (roduct, iger in ,ran and oter# fraction
DON and CEA % ige#t in ,ran and lo;e#t in te flour DON and CEA % ige#t in ,ran and lo;e#t in te flour
Fu+oni#in
1umonisin concentration )ugG/g.
Corn fraction IE I) I7
Hhole corn +;69 <+: +;,
-rits +>; >= +9
ICI grade flour >;8 +,, 6:
Corn meal +68 ;, ,8
-erm J bran 6,+9 ,9+9 66<
*roggi et al 7, ,99,
4ET MI''IN!
PRODUCE STARCH AT EXTREEMELY LOW LEVEL OF MYCOTOXINS
AFB : were lost in steep liquor and concentrated in the ger
ZEA,NIV, DON ; similar result (NIV and DON soluble in steep liquor)
B1 2.59 Zea mays
B2 0.45 Feed (ppm)
Cleaning
Corn step B1 0.02
Steeping liquor
Milling Germ B1 0.15
B2 0.05
Fraction & Fiber B1 0.45
Separation B2 0.25
Gluten B1 0.72
B2 0.28
Dry Sacharification
B1 ND Corn Starch Convet into Corn oil
B2 ND Sugar
Distribution of fumonisins in wet milling process
(amimura, %7,+===
IAKIN!, ROASTIN!, FR$IN! - Scott EB27, Rei## EB12,
Su,irade EBBF, Scuda+ore et al?, )**7, Sa+ar et al, )**E? Ca#telo et al?, )**E
AFIE % destroyed during dough fermentation lost during
ba/ing ) up to ,;F.
almost complete destruction in fried snac/
)pretreatment ? >F NaO%, autocla4ing and
frying +; min at +=:KC. frying +; min at +=:KC.
OTA % 'ittle lost during ba/ing )cacao for fla4orL.
DON % The maximum reduced during dough fermnt7 6:F
FUMONISIN% "educed about ,8F )ba/ing at ,99KC for ,9 min.
No significance lost spi/ed corn fried at +<9KC,
degradation occurred started at +89KC
1rying corn chips at +=9KC for +; min7:<F
(ppb)
Raw peanut B1 0.24-2.38
Shelled peanut B2 Tr- 0.26
Steep tankes G1 ND-0.19
B1 0.034 G2 ND-Tr
B2 0.012 Steep water
Steep peanut B1 0.73
B1 0.66 B2 0.15
B2 0.09 Coat splitting
G1 0.16 Seed coat Peanut kernel B1 0.50
G2 0.02 Sorting B2 0.11 G2 0.02 Sorting B2 0.11
B1 0.64 Damage kernel kernel (normal) ND
B2 0.06 Frying
Fry kernel B1 Tr
B1 0.42 Frying oil
B2 Tr Product ND
Distribution of aflatoxins in butter-peanut process
Nixtamalization )alkaline cooking and heating)
Technology used for preparing traditional foods from corn (tortillas,
for example), significantly reduced the concentration of aflatoxin.
(Ulloa-Sosa and Shroeder, 1969; Price and Jogersen, 1985;
Elias-Orozco et al., 2002) UP TO 79% REDUCED
NIV, DON : reduced 60-100% in aqueous bicarbonate at 10-50%
COOKIN! AND CANNIN!
NIV, DON : reduced 60-100% in aqueous bicarbonate at 10-50%
by weight (Lauren and Smith, 2001)
reduced 40% and 60% boiled noodle at 98C for 10 min
ZEA : not reduced by 12 days at 110C heating after treatment
with bicarbonate (Kamimura, 1980)
T2 toxin: 30% reduced in the same heating (Kamimura,,1980)
FUMONISIN : 11-15%reduced; 90% after alkaline steeping
OTA : stable during cooking; citrinin citrinin H2
EXTRUSION
A system in which raw materials are passed through continuous
processing machinery within which compressed and sheared at
elevated temperature and pressure resident time is short
MYCOTOXIN !"OC$ $11$CT "$11
&1*+ C &1*, $xtrusion at +;9C, spi/ed corn sur4i4e Martine5, +=8=
dough
&1*+ Corn flour )J ;9MgG/g. ,;F reduction Ca55aniga et al7, ,99+ &1*+ Corn flour )J ;9MgG/g. ,;F reduction Ca55aniga et al7, ,99+
DON Corn four )J; mgG/g. =;F reduction
&1*+ C &1*, $xtrusion rice flour )+690,99CB
scre2 speed +>9 rpmB moist7 ,9F. <;F lost Camargo et al7, +=8=
OT& ;9F moistB +;9C partially decomposed *oudra et al7,+==;.
DON @uite stable Holf0%all et al7, +===
3$& 4arried 2ith scre2 configuartion :;08>F reduction "yu et al7,+===
1#MONIIN extrusion of dry mill product >90=9F reduced aunders et al7,,99+
1M*+ C 1M*, coo/ing, extrusion, gelatini5ation >90;;F reduction Meuster, ,99+
1M*+ C 1M*, corn fla/es, extrusion and roasting :90<9F lost Degirolamo, et al7,,99+
IRE4IN! 9
A'CO/O'IC
FERMENTATION
DECOM&OSED 9
C/AN!E %
OTA
TRIC/OT/ECENE TRIC/OT/ECENE
ISO TRIC/OT/ECENE
&ATU'IN
"$11 ? cott et al%+==,B ch2ar5 et al.%+==;
DID NOT C/AN!E DON, FUMONISIN
T/E CONTRIIUTION OF SE:ERA' FOOD &ROCESSIN! ON
REDUCTION OF M$COTOXINS
E? EDII'E OI' &ROCESSIN! Aflato<in ;ere accu+ulated into P#oa(
#toc>Q during neutrali=ation (roce## ? &eanut oil ;itout (urification
are ar+full e<a+(le % aflato<in in Pin#tant noodleQ
)? 4/EAT &ROCESSIN! +ycoto<in# -e#(c?fu+oni#in. are accu+ulated
in ,ran, ger+ and #tee(ing ;ater, #+all (art #tilll in final (roduct#
7? CORN STRAC/ &ROCESSIN!+ycoto<in# are accu+ulated in ger+,
,ran and Pcorn #tee( liRuor Pfor feeding? In ca#e of P+arningQ
(roce##ing, reduction of aflato<in# occurred during ,oiling in li+e
#olution
@? &EANUT IUTTER &ROCESSIN! aflato<in found in #eed coat,
#tee(ing ;ater, frying oil and trace are re+ain in final (roduct
5? CIDER &ROCESSIN! % #afe for (atulin
F? COFFEE &ROCESSIN! %
OTA re+o"e ,y initial cleaning, reduction in
roa#ting -EF3 fro+ #tarting concentration .
AFI B73 lo#t during roa#ting
1? IE:ERA!E % 1? IE:ERA!E %
(art of +ycoto<in# ;ere re+o"ed ,y
#orting te decayed fruit?
&atulin @*3 re+o"ed ,y clarification
(rocedure
NIXT&M&'I3&TION ? Not effecti4e for 3$&, 1#MONIIN
and OT&
N$#T"&'I3&TION ? $ffecti4e for &1*+
"O&TIN- ? reduce part of mycotoxins , almost
remo4ed &1* by lime coo/ing as remo4ed &1* by lime coo/ing as
pretreatment
*&(IN- ? "educe relati4ely high part of
mycotoxins, except OT&
1$"M$NT&TION ? destroyed by binding, en5ymatic
detoxification, other reactions
CONCLUSION
Prevention of mycotoxins contamination is not always
possible?
Procedure suitable for destruction of multiple
mycotoxins still limited
Almost impossible to establish physical and chemical Almost impossible to establish physical and chemical
treatment would be useful for destruction
mycotoxins simultaneously
Biological methods more promising for detoxification
Be possible to introduce modification to commercial
process to get the reduction of mycotoxins in
retail product