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Comprehensive
Review of
Patulin Control
Methods in
Foods
Matthew M. Moake, Olga I. Padilla-Zakour,
and Randy W. Worobo
ABSTRACT: The mycotoxin, patulin (4-hydroxy-4H-furo [,!c" pyran-![#H"-one$, i% produced &y a num&er of fun'i com-mon to fruit-
and e'eta&le-&a%ed product%, mo%t nota&ly apple%) *e%pite patulin+% ori'inal di%coery a% an anti&iotic, it ha% come under heay
%crutiny for it% potential ne'atie health effect%) Studie% ine%ti'atin' the%e health effect% hae proed inconclu%ie, &ut there i% little
dou&t a% to the potential dan'er inherent in the contamination of food product% &y patulin) The dan'er po%ed &y patulin nece%%itate% it%
control and remoal from food product%, creatin' a demand for handlin' and proce%%in' techniue% capa&le of doin' %o, prefera&ly at
lo co%t to indu%try) .ith thi% &ein' the ca%e, much re%earch ha% &een deoted to under%tandin' the &a%ic chemical and &iolo'ical
nature of patulin, a% ell a% it% interaction ithin food% and food production) .hile pa%t re%earch ha% elucidated a 'reat deal, patulin
contamination continue% to &e a challen'e for the food indu%try) Here, e reie in depth the pa%t re%earch on patulin ith an empha%i%
upon it% influence ithin the food indu%try, includin' it% re'ulation, health effect%, &io%ynthe%i%, detection, uantification, di%tri&ution
ithin food%, and control, durin' the ariou% %ta'e% of apple /uice production) 0inally, 1ey area% here future patulin re%earch %hould
focu% to &e%t control the patulin contamination pro&lem ithin the food indu%try are addre%%ed)
Introduction
Patulin is a mycotoxin produced by a number of fungi common to
fruit- and vegetable-based products, most notably apples. Ap-ples
are the 3rd most important fruit crop in the United States after citrus
fruits and grapes, with !" of apples being used for #uice and other
processed products $US%A &!!&'. Patulin contamina-tion within
apple products poses a serious health ris( to consum-ers,
particularly children whom have been shown by a US%A sur-vey to
consume increased levels of apple products during the )st y of life
$*. g+(g body weight+d', compared with adults $) g+(g bw+d'
$Plun(ett and others )&', placing them at increased ris( for
patulin toxicity. he health ris(s posed by patulin necessitate its
control and removal from apple products, creating a demand for
food-processing techniues capable of doing so, preferably at low
cost to industry. /ere, we review past research devoted to the
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understanding and control of patulin, with an emphasis upon pat-
ulin0s influence within the food industry.
History and Regulation
Patulin $-hydroxy-/-furo 13,&c2 pyran-&1*/2-one', igure ), is
a water-soluble lactone )st isolated as an antibiotic during the
MS 20040335 Submitted 5/21/04, Revised 8/17/04, Accepted 10/18/04.
Authors are with Dept. of Food Science and Technology, New York State
Agricultural Experiment Station, Cornell Univ., Geneva, NY 14456-0462.
Direct inquiries to author Worobo (E-mail:[email protected]).
)!s $Stott and 4ullerman )56'. 7wing to co-discovery of the
compound by various groups, it has historically been (nown by
names such as clavacin $Anslow and others )3', expansine
$8an 9ui#( )3:', claviformin $;hain and others )&', clavatin
$4ergel and others )3', gigantic acid $Philpot )3', and
myo-cin ; $%e
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Patulin control in foods . . .
Table 1The health effects of patulin
Acute symptom
Source
Agitation, convulsions, dysponea, pulmonary congestion,
Escoula and others 1977; Hayes and others 1979
edema, hyperemia, GI tract distension
Nausea
Walker and Wiesner 1944
Epithelial cell degeneration, intestinal hemorrhage
Mahfoud and others 2002
Intestinal inflammation
McKinley and Carlton 1980a, 1980b; McKinley and others
1982; Mahfoud and others 2002
Ulceration
Escoula and others 1977; Hayes and others 1979; McKinley and Carlton 1980a,
1980b; McKinley and others 1982; Mahfoud and others 2002
Chronic symptom
Source
Genotoxic
Mayer and Legaror 1969; Korte 1980; Thust and others 1982; Lee and Roschenthaler
1986; Roll and others 1990; Hopkins 1993; Pfeiffer and others 1998
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Neurotoxic
Hopkins 1993
Immunotoxic
Hopkins 1993; Wichmann and others 2002
Immunosuppressive
Wichmann and others 2002
Teratogeneic
Ciegler and others 1976; Roll and others 1990
Carcinogenic
Dickens and Jones 1961; Oswald and others 1978
Cellular level effect
Source
Plasma membrane disruption
Riley and Showker 1991; Mahfoud and others 2002
Protein synthesis inhibition
Hatez and Gaye 1978; Miura and others 1993; Arafat and Musa 1995
Transcription disruption, translation disruption
Moule and Hatey 1977; Arafat and others 1985; Lee and Roschenthaler 1987
DNA synthesis inhibition
Cooray and others 1982
Na-coupled amino acid transport inhibition
Ueno and others 1976
Interferon- production inhibition
Wichmann and others 2002
RNA polymerase inhibition
Moule and Hatey 1977
Aminoacyl-tRNA synthetases inhibition
Arafat and others 1985
Na-K ATPase inhibition
Phillips and Hayes 1977, 1978; Riley and Showker 1991
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Muscle aldolase inhibition
Ashoor and Chu 1973
Urease inhibition
Reiss 1977
Loss of free glutathione
Burghardt and others 1992; Barhoumi and Burghardt 1996
Protein crosslink formation
Fliege and Metzler 1999
Protein prenylation inhibition
Miura and others 1993
Fhile 6! g+9 is now the norm for patulin regulation, several countries
have set even lower limits for patulin at &6 to 36 g+9 $van Dgmond
):'. =n con#unction with these maximum content standards, a #oint
ood and Agriculture 7rgani?ation-Forld /ealth 7rgani?ation
$F/7' expert committee established a provi-sional maximum daily
inta(e of !. g+(g body weight for patulin $F/7 )6'. he United
States has been much slower to set reg-ulation on patulin, but today
the U.S. ood and %rug Administra-tion limits patulin to 6! g+9 in
single-strength and reconstituted apple #uices $US%A &!!'. he
limitation of these regulations to apple #uice and apple #uice
concentrate was li(ely based upon the fact that, at the time, only
apple #uice and cider had been
Figure 1The structure of patulin !"#hydro$y#"H#furo %&'(c) pyran#
(%*H)#one+
found to be naturally contaminated by patulin $F/7 )!'.
Fhile this fact has since been disproved $see below, Patulin
Fith-in oods', apple #uice and cider remain the ma#or source of
hu-man patulin consumption.
Health ,ffects
Assessment of the health ris(s posed by patulin to humans is based
upon a wide number of studies during the past 6!-plus years that
implicate a number of acute, chronic, and cellular level health effects
as summari?ed in able ). inley and ;arlton ):!a, ):!bB
c>inley and others ):&B ahfoud and others &!!&'. ;hronic
health ris(s of patulin consumption can include neuro-toxic,
immunotoxic, immunosuppressive, genotoxic, teratogenic, and
carcinogenic effects $%ic(ens and Eones )*)B ayer and 9egaror
)*B ;iegler and others )5*B 7swald and others )5:B >orte
):!B hust and others ):&B 9ee and
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)56B 9ynen and others )5:'. he
gene encoding for *-SA has been
cloned and characteri?ed from P.
patulum and P. urticae $4ec( and
others )!B Fang and others ))'.
=nactivation of *-SA synthetase has
been shown to be the )st limitation on
patu-lin production $Ceway and
@aucher ):)'. *-SA synthetase loss
is a selective process because the
highly similar fatty acid synthetase of P.
urticae $9ynen and others )5:' is
stable under the same reaction
conditions that inactivate *-SA
synthetase $9am and others )::'.
his finding is further verified by
studies in which treatment of *-SA
synthetase reaction mixtures, contain-
ing Cicotanimide Adenine %inucleotide
Phosphate $CA%P/' co-factor, acetyl-
;oA, and malonyl ;oA, with the
reducing agent, dithiothreitol, and
proteinase inhibitor,
phenylmethylsulfonyl fluo-ride,
stabili?ed *-SA synthetase. his
suggests proteolysis and
conformational integrity play a role in
the regulation of *-SA synthetase
$9am and others )::'.
he next stage of patulin biosynthesis
involves the conversion of *-SA into m-
cresol via the activity of *-SA
decarboxylase $9am and others )::'.
-cresol is then converted into m-hy-
droxyben?yl alcohol by m-cresol &-
hydoxylase $urphy and 9ynen )56'.
he next step in patulin0s biosynthetic
pathway is debated among & main
mechanisms. 4oth agree that m-hydroxy-
ben?yl alcohol is eventually converted to
gentisaldehyde $orrest-er and @aucher
)5&B Lamir ):!', however the
intermediary be-tween these &
compounds is believed to be either
gentisyl alcohol $Se(iguchi and others
):3B =i#ima and others ):*' or m-hydrox-yben?aldehyde $Se(iguchi and
others ):3'. Some studies have
suggested that both are possible, with m-
hydroxyben?aldehyde being favorable
$@aucher )56', whereas others believe
that m-hydroxyben?aldehyde is not
converted to gentisaldehyde but rather to
m-hydroxyben?oic acid $urphy and
9ynen )56'. =n the &nd case, m-
hydroxyben?yl alcohol dehydrogenase
converts m-hydroxyben?yl alcohol into m-
hydroxyben?aldehyde $@auch-er )56B
urphy and 9ynen )56'. 4oth this
en?yme and m-cresol &-hydroxylase have
been shown to reuire oxygen and
CA%P/ to function $urphy and 9ynen
)56'.
7nce gentisaldehyde has been formed, it
is then converted to isoepoxydon,
phyllostine, neopatulin, D-ascladiol, andfinally to patulin $Se(iguchi and @aucher
)55, )5:B Se(iguchi and @au-cher
)5B Se(iguchi and others )5, ):3'.
he conversion of isoepoxydon to
phyllostine is accomplished via an CA%P-
depen-dent isoepoxydon dehydrogenase
$Se(iguchi and @aucher )5'.
;onversion of neopatulin to D-ascladiol is
accomplished through a reduction by
CA%P/. he product of this reaction, D-
ascladiol, is then either oxidi?ed to patulin
or nonen?ymatically transformed to its
isomer L-ascladiol $Se(iguchi and others
):3'. he biosynthetic pathway ofpatulin is summari?ed in igure &.
.etection and /uantification
he established limit of 6! g+9 in the
United States as the maxi-mum patulin
level allowed in fruit products has
provided an in-centive for the
development of faster and more specificanalytical methods with lower detection
limits. A comprehensive review of the
development of thin-layer gas and liuid
chromatographic methods for the
detection and confirmation of identity of
patulin has been previously published by
Shephard and 9eggott $&!!!'.
he most common method currently used
to uantify patulin in fruit products is high-
performance liuid chromatography
$/P9;' with ultraviolet $U8' detection.
his is the official method adopted by
A7A; =ntl. for apple #uice $method
6.)!' with a detection limit of 6 g+9. he
#uice is extracted 3 times with ethyl
acetate and cleaned up by liuid-liuid
extraction with a ).6" sodium car-bonate
solution. he ethyl acetate extract is dried
with anhydrous sodium sulfateB the
solvent is then evaporated, normally
under ni-trogen, and the dried residue is
dissolved with acidified water $p/
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Patulin control in foods . . .
by addition of
acetic acid'.
his prepared
extract is ready
for /P9;
analysis. he
recommendedliuid
chromatograph
y $9;' systems
include
analytical
reversed-phase
9; columns
such as
octadecylsilane
fully end-
capped with 6
m particle
stationaryphase, )& to &6
nm pore si?e,
carbon loading
of )&" to )5",
and a U8
detector set at
&5* nm,
although a
photo diode
array detector
is preferred to
aid in the
presumptiveidentification of
the patulin
pea(. he
system can be
run isocratically
at ) m9+min us-
ing 3" to )!"
acetonitrile in
acidified water
$!.!6 parts
per vol-ume
perchloric acid
*!"' as long
as patulin
separates from
6-hy-
droxymethylfurf
ural $/', a
common
compound
found in apple
#uice that elutes
#ust before
patulin. or
cloudy apple
#uice and ap-
ple puree, a
collaborative
study of )&
participants
from Duropean
countries was
recently
conducted tovalidate the
effectiveness of
this 9;
procedure for
patulin
determination
with a slight
modifica-tion.
Prior to the
ethyl acetate
extraction, the
samples were
treated withpectinase
en?ymes and
held overnight
at room
temperature or
for & h at ! M;
and then
centrifuged at
6!! N g for 6
min. 4ased on
the results, the
method is
recommendedfor patulin at
greater than 6!
g+9 in cloudy
apple #uice and
purees $ac-
%onald and
others &!!!'.
he
simultaneous
uantification ofpatulin and /
in apple #uice by
reversed-phase
/P9; has been
reported by
@o(men and
Acar $)'.
he method
developed uses
a 6- m ;):
analytical
column $)6! N
mm', a
photodiode
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array detector
operating at &6!
to 3!! nm, and
a mobile phase
of water-
acetonitrile $J)
v+ v' at ).!
m9+min. Sample
preparation
followed the
official method
describedearlier.
;omplete
separation of
/ and patulin
was achieved in
less than min
at detection
limits of less
than !.!) g+9
and less than 6
g+9 respectively.
he /P9;+U8
procedure is
routinely used
for uantitative
deter-mination
of patulin in
apple products,
but methods to
confirm the
presence of
patulin usually
include more
specificdetection tech-
niues such as
mass
spectrometry
$S' after 9; or
gas chroma-
tography $@;'
separations. or
@;-S
analysis, patulin
is detect-
ed as its
trimethyl silyl
derivative
$S-patulin'
with electron
ion-i?ation
$
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assays using
)3;-labeled
patulin as the
internal
standard. 7ne
method used
9;+S in neg-
ative
electrospray
ioni?ation mode
without
derivati?ation,
while the &nd
procedure
utili?ed high
resolution gas
chromatography
+ high resolution
mass
spectrometry
$/
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relevant en#2ymes involved in the
biosynthesis of patulin !3aucher
14567 Murphy and 8ynen 14567
e9iguchi and 3aucher 14547
e9iguchi and others 14:&7 Ii;ma
and others 14:*7 Priest and 8ight
14:4+0
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Patulin-production within fruits,
vegetables, and their products has been
investigated and often appears to be
dependent on such factors as water
activity $aw', temperature, p/, and other
chemical characteristics intrinsic to fruits
$Sommer and others )5B
Cortholt and others )5:B c;allum and
others &!!&'. p/ and patulin production
have been shown to be inversely related,
with patulin being unstable at high p/
$c;allum and others &!!&'.
emperature has been shown to affect
pathogen growth and, to a greater extent,
the production of patulin $c;allum and
others &!!&'. Patulin production has
been observed at all temperatures
permitting P. expansum growth,
encompassing an approximate range of !
to 3! M; $Sommer and others )5'. B.nivea has been shown to grow faster at
temperatures of 3! and 35 M; while patu-
lin production was highest at &) M;
$
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and Australia $Sommer and others
)5'.
Cumerous studies around the world
have examined the extent and degree
to which apple products have been
contaminated by
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Patulin control in foods . . .
patulin. =n
Fisconsin, &3
of ! roadside
apple #uices
were found to
contain
between )! to36! g patulin+9
$4rac(ett and
arth )5a'.
A &nd study
showed that :
of )3
commercial
apple #uices
tested
contained
between
and 3! g
patulin+9 #uice$Fare and
others )5'. A
ur(ish study
showed &)6 of
&)6 apple #uice
concentrates
examined had
between 5 and
356 ppb patulin
with 3" being
above the 6!-
ppb
internationalstandard
$@o(men and
Acar ):'.
inally, a )*
to ): study
in South Africa
showed that 6
of && #uices
sampled
contained
between )!
and 6 ppb
patulin, and
&" of infant
apple products
showed 6 to &!
ppb $4rown
and Shephard
)'. A
summary of
patulin con-
tamination
within foods is
given in able
&.
Control
.uring =pple
Harvest'
Processing'
and torage
=n Corth
America, apple
#uice is typically
a byproduct
pro-duced from
culled apples
unfit for other,
higher uality
and higher
profit, purposes
$
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efforts, and $'
adherence to
/a?ard
Analysis
;ritical ;ontrol
Points
principles
$9ope?-@arcia
and Par( ):B
Par( and
others )'.Dxisting and
developmental
preventive
measures
during
preproduction
are based upon
fruit uality and
facility
sanitation
measures. he
uality of fruit
resulting fromharvesting is
the )st step in
controlling
patulin levels.
Fith the
highest uality
hand-pic(ed
fruit being used
for direct-for-
retail sale,
processed
apple products
usually areproduced from
mechani-cal
harvest,
windfalls,
insect-
damaged, or
culled fruit.
4ruises, s(in
brea(s, and
other physical
damage within
these apples
pro-vide a
perfect entry for
P. expansum
and other
patulin-
producing
species into the
fruit. Studies
have examined
the effect of
fruit uality and
harvest method
on the patulin
content of the
result-ant
#uices. =n )
study, patulin
was
undetectable in
5 cultivars of
tree-pic(ed
cider, whereas
it was detected
between !.&
and 35 g+9 in cultivars of
ground-harvest
cider $Eac(son
and others
&!!3'. any of
the patulin
control
measures
suggested by
the Eoint
A7+F/7
ood
StandardsProgramme are
based upon the
careful
selection of
fruits as based
upon good
agricul-tural
practice
$;7%DQ &!!&'.
/owever, as
indicated by a
Cew Gor( State
/udson 8alley9ab study that
showed around
&!" of bagged,
in-store, retail
apples to
contain
consumer-
visible blue-rot
$
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processing,
including
washing,
sorting, and
pac(aging,
poses a &nd
means of both
fungal control
and
contamination
ali(e. Fashing
with high-pressure water
has been shown
to reduce patulin
levels within
apple #uice by
&)" to 6"
$Acar and others
):'. A &nd
study showed
that washing of
ground-
harvested
apples resulted
in a )!" to
)!!" patulin re-
duction,
depending on
initial patulin
level and
washing
treatment
$Eac(son and
others &!!3'.
/owever, these
same washes
can also serve
as a source ofcontamination.
;ontaminated
bins, storage
rooms, drencher
washes, drying
brushes after
apple wash, and
other steps
within the
processing cycle
can all provide a
source of fungal
inoculum cycling
in poorlysaniti?ed setups.
Prevention
methods aimed
at cleaning and
sterili?ing
storage and
processing
facilities
routinely and in
between
seasons are
being mapped
out, but have not
yet been fully
developed. Plus,
the older,
complicat-ed
design of many
pac(ing-house
and processing
euipment in-
hibits the ability
to effectively
saniti?e. Dven if
effective
strategies are
successfully
mapped out,
the inherent
variability in
apple han-dling
facilities will
reuire
customi?ation
of sanitation
methods for
each operation
$
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stem-end in-
fected blue-rot
began to
appear with
increasing
freuency.
9ong-term,
controlled
atmosphere
storage has
now beenshown to allow
the slow growth
and stem-
based invasion
of fungi into
apples
$
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$
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their
effectiveness
against fungal
spores.
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Pierson and others 1971; Buchanan and others 1974; Sommer and others 1974
Apricots
Harvey and others 1972; Buchanan and others 1974; Sommer and others 1974
Persimmons
Sommer and others 1974
Strawberries
Andersson and others 1977; Frank and others 1977; Arici and others 2002
Nectarines
Harvey and others 1972
Raspberries
Andersson and others 1977; Arici and others 2002
Black mulberries
Arici and others 2002
White mulberries
Lingon berries
Andersson and others 1977
Peaches
Harvey and others 1972; Buchanan and others 1974; Andersson and others 1977; Frank
and others 1977
Plums
Harvey and others 1972; Buchanan and others 1974; Andersson and others 1977
Tomatoes
Greengages
Frank and others 1977
Bananas
Blueberries
Akerstrand and others 1976; Andersson and others 1977
Black currants
Andersson and others 1977
Almonds
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Pecans
Jiminez and others 1991
Peanuts
Hazelnuts
Foods contaminated with patulin Source
Apple juice
Lovett and others 1974; Sommer and others 1974; Frank and others 1977; Scott and others
1977;
Brackett and Marth 1979a; Prieta and others 1994; Rychlik and Schieberle 1999;
Leggott and Shephard 2001; Ritieni 2003
Apple-acerola juice
Rychlik and Schieberle 1999
Pear juice
Ehlers 1986
Grape juice
Harwig and others 1978; Rychlik and Schieberle 1999
Sour cherry juice
Blackcurrant juice
Rychlik and Schieberle 1999
Orange juice
Pineapple juice
Ake and others 2001
Passion fruit juice
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Apple cider
Brackett and Marth 1979; Wheeler and others 1987; Leggott and Shephard 2001
Apple puree
Leggott and Shephard 2001; Ritieni 2003
Corn
Lin and others 1993
Strawberry jam
Blackcurrant jam
Jelinek and others 1989
Blueberry jam
Baby food
Prieta and others 1994; Leggott and Shephard 2001; Ritieni 2003
Cheddar cheese
Bullerman and Olivigni 1974
Barley Malt
Lopez-Diaz and Flannigan 1997
Wheat Malt
Bread
Reiss 1972, 1976
Countries with apple and
juice contamination
Source
Canada
Scott and others 1972; Harwig and others 1973b; Sommer and others 1974
England
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Brian and others 1956; Sommer and others 1974
New Zealand
Walker 1969
United States
Sommer and others 1974; Ware and others 1974
South Africa
Leggott and Shephard 2001
Sweden
Josefsson and Andersson 1976
Turkey
Gokmen and Acar 1998
Brazil
de Sylos and Rodriguez-Amaya 1999
Austria
Steiner and others 1999a
Belgium
Tangi and others 2003
Australia
Sommer and others 1974
France
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Patulin control in foods . . .
standard apple
#uice production
steps are
centered on 3
areas. he )st
of these
involves theuality of the
fruit and
processing of
this fruit, prior
to pressing. As
previously
mentioned,
processed
apple products
often utili?e
lower uality
fruit that is
unsuitable fordirect mar(et
retail.
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necessitating
the need to
re#ect entire
#uice loads of
apples. =f
reliant on this
method, small-
scale
producers who
cannot afford
these losseswill be forced to
sort by hand,
in-creasing
costs to the
point that many
may cease
operating
$
8/20/2019 SIKLUS BIOSINTESIS PATHWAY
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reduce patulin
levels by :"
and 55",
respec-tively.
/owever, these
methods ma(e
the removed
ca(e and+or fil-
ter potentially
highly toxic and
unfit for any
further use, suchas often is done
with #uice
sediments in
animal feed
$4issessur and
others &!!)'.
his could
represent a loss
of income for
many #uice
producers.
4atch
absorption with
synthetic
polymers has
also been
investigated
$;anas and
Aranda )*',
as has en?yme
treat-ment with
pectinase
en?ymes used
to brea( down
the pectin coat
surrounding
protein particles,allowing
aggregation and
sedimen-tation
of protein
particles and, in
the case of
patulin
contamina-tion,
their associated
patulin adducts.
his method has
resulted in a
53" decrease in
patulin contentwithin #uice
$4issessur and
oth-ers &!!&'.
he 3rd #uice
production
process
capable of
reducing
patulin
levels is the
pasteuri?ation
process. 7f the
3 processes
mentioned thus
far, this is by far
the least
effective.
8/20/2019 SIKLUS BIOSINTESIS PATHWAY
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to reduce patulin
levels by ):.:"
$Fheeler and
others ):5'.
Dv-idence also
shows that
patulin is
nonvolatile and,
upon distilla-tion
production of
apple aroma,
patulin remainswithin #uice con-
centrate $>ryger
&!!)'. inally,
not only is the
pasteuri?ation
pro-cess unable
to significantly
reduce patulin
levels, it often
fails to fully
remove heat
resistant patulin-
producing fungi,
such as B. nivea
and B. fulva
$7ugh and
;orison ):!',
allowing for
poten-tial
continued
production of
patulin within the
finished #uice.
As a final stageof the production
process, studies
have exam-ined
the effects of
storage on
patulin content.
ixed studies
show either a
decrease $Scott
and Somers
)*:B /arwig
and others
)53a' or no
change
$Pohland and
Allen )5!B
Legota and
others )::' of
patulin levels in
apple #uice with
refrigerated
storage. Similar
studies within
grape #uice have
shown stability
$7ugh and
;orison ):!' or
an approximate
6!" decrease
$Scott and Som-
ers )*:' of
spi(ed patulin
levels in grape
#uice after 6 w(.
Control
Postproducti
on
Filtering and
adsorption
A number of
studies have
been devoted to
the removal of
patu-lin from
#uice through the
use of
adsorption
filters, columns,
and agitation
treatments using
carbon-based
material.
Agitation with &!
mg+m9 activated
charcoal
followed byfiltration through
a !-or *!-mesh
charcoal column
reduced a 3!-
g+m9 patulin
solu-tion to
below
detectable
levels. urther,
use of 6 mg+m9
charcoal in
agitation was
able to reduce
patulin to belowdetectable levels
in naturally
contaminated
cider. /owever,
color loss was
mar(ed-ly
present in this
resulting #uice
$Sands and
others )5*'. =n
a &nd study, !,
!.6, ).!, ).6,
&.!, &.6, and 3
g+9 activated
8/20/2019 SIKLUS BIOSINTESIS PATHWAY
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charcoal was
added to
naturally
contaminated
apple #uice
containing *&.3
ppb patulin.
Samples were
mixed for !, 6,
)!, &!, and 3!
min. hree
grams+literactivated
charcoal was
found to be most
effective with a
time period of 6
min. ;learness
of #uice
increased, color
of #uice
decreased, and
small decreases
in fumaric acid,
p/, and M4rix
were also seen
$>ada(al and
Cas &!!&'. =n
another study,
ultrafine
activated carbon
was bound to
granular uart?
producing a
com-posite
carbon
adsorbent
$;;A'. ;olumns
with varyingamounts of ;;A
were prepared
and )! g+m9
patulin were
filtered through
at ) m9+min.
ifty percent
brea(through
values for
columns with
).!, !.6, and
!.&6 g ;;A
were )35.6,
3:.6, and ).
g, respectively
$/uebner and
others &!!!'.
=n a th study
designed to
compare theeffects of
different car-bon
activation
methods, steam
activated carbon
C7
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;
8/20/2019 SIKLUS BIOSINTESIS PATHWAY
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through the addition of sulfur dioxide to
the hemiacetal ring of patulin, forming a
carbonyl hydroxysulfonate. he &nd
reaction is thought to be irreversible and
to occur due to an opening of the lactone
ring structure at one
of the double bonds $4urroughs )55'.
A th study found the sul-fur dioxide
inactivation of patulin to be reversibleor irreversible de-pending on p/
$Steiner and others )b'.
Sulfur compounds common to
biological systems and fre-uently
associated with patulin toxicity, such as
glutathione, cys-teine, and
thioglycolate, are believed by many to
produce biologi-cally inactive products
when reacted with patulin $;avallito
and 4ailey )B @eiger and ;onn
)6B en?ie and others )5'.
ur-ther, other mycotoxins such as
aflatoxins 4), @), 4&, and @& $aeba
)::' as well as ?earalonone $%oyle
and others ):&' have been shown to
be effectively degraded by o?one.
=n all of these cases, preliminary
evidence is promising. /owev-er,inadeuate research has been done to
examine the efficacy and full functional
range of applicability for the treatment.
urther-more, the reaction mechanisms
for few if any of the chemical
detoxification treatments are fully
understood, and both the reac-tion
products and their respective toxicities
are still un(nown and must be
determined prior to use. he regulatory
approval for some of the proposed
chemical treatments such as
ammoniation and potassium
8/20/2019 SIKLUS BIOSINTESIS PATHWAY
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permanganate must be sought prior to
use in the food industry.
-iological control
4iological methods of patulin control
result largely from the ob-
16 ;7P
8/20/2019 SIKLUS BIOSINTESIS PATHWAY
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Patulin control in foods . . .
servation that
patulin is almost
always
completely
degraded dur-
ing yeast
fermentation.
4esides being
uite successful,
this meth-od is
much better
understood
compared with
other
decontamina-
tion methods.
Approximately
!" of patulin
can be removed
during yeast
fermentation$4urroughs
)55'. =n )
study, * of :
yeast strains
reduced patulin
levels to below
detectable
levels, while all
: strains
resulted in a
" or better
decrease in total
pat-ulin content.
;ontrol #uice, onthe other hand,
stored for an
eual amount of
time $& wee(s',
had only a )!"
reduction
$Stinson and
others )5:'. =n
a &nd study,
yeast
fermentation
reduced patulin
levels
completely after
& w(. his same
study also
showed that
patulin levels
failed to
decrease
significantly in
#uices that had
been yeast
fermented and
then filter
sterili?ed to
remove yeast,
suggesting that
active yeast,
and not their
byproducts,
were re-uired
for the reduction
$/arwig and
others )53a'.
reatments of
patulin along
with
cyclohexamide,
a protein
synthesis
bloc(er of yeast,
completely
bloc(ed protein
synthesis and
prevented the
detoxification of
patulin. Addition
of
cyclohexamide 3h after pat-ulin
addition,
however,
resulted in a
reduced, but
continued, rate
of patulin
degradation,
suggesting that
the proteins
synthesi?ed
within the 3-h
window were
catalyticallyactive against
patulin and did
not #ust bind it
up in adduct
formation
$Sumbu and
others ):3'. A
later study
showed that 3
strains of
Saccharomyces
cere-visiae
reduced patulin
levels during
fermentive
growth but not
aerobic growth.
his reduction
resulted in the
production of &
ma-#or productsJ
D-ascladiol,
patulin0s
immediate
biosynthetic
precur-sor, and
its isomer L-
8/20/2019 SIKLUS BIOSINTESIS PATHWAY
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ascladiol. hese
& products were
also seen in the
treatment of
patulin with the
reducing agent
sodium borohy-
drate $oss and
9ong &!!&'. D-
ascladiol is itself
a mycotoxin
$Su-?u(i andothers )5)',
which has
reduced toxicity
compared with
patulin and also
reacts with
sulfhydryl-
containing
compounds
$Se(iguchi and
others ):3'.
Fhile effective,
biological
control with
yeast is limited
to prod-ucts
that can be
fermented.
urthermore,
yeast are
themselves
sensitive to
patulin, and atconcentrations
greater than
&!! g+ m9,
yeast have
been shown to
be completely
inhibited,
prevent-ing
fermentive
detoxification
$Sumbu and
others ):3'.
Co re-searchhas been done
to examine the
potential use of
other fer-
menting
microbes, such
as lactic acid
bacteria, in
decreasing pat-
ulin content
within #uices.
Similar
reducing
en?ymes and
environ-ments
produced by
these bacteria
may very well
be able to de-
grade patulin.
inally, no
research has
investigated the
direct en-?ymatic
degradation of
patulin.
8/20/2019 SIKLUS BIOSINTESIS PATHWAY
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been performed
specifically on
patulin $%oyle
and others
):&B 8alletrisco
and others
)!'.
Patulin
Production
in the Final
Product
9imited wor(
has been done
to examine the
potential growth
and production
of patulin by
fungi in stored,
finished product
#uices. =t is well
(nown that
many
Aspergillus and
Byssochlamys
spp. molds,
many of which
produce patulin,
are heat
resistant and
can survive the
pasteuri?ation
processes usedin #uice and ci-
der production.
urthermore,
many of the
same patulin-
produc-ing
molds have also
been shown to
grow and
produce patulin
at
standard cold
storage
temperature
down to !M;
$Sommer and
others )5'.
a(en together,
these & facts
suggest that
finished apple
#uice and cider
could very well
support the
continued pro-
duction of
patulin by fungi.
=f this is the
case, any
single
treatment
patulin control
method, such
as physical
adsorption,would po-
tentially be
moot due to
production of
patulin by
active fungi
postprocessing.
Studies must
be done to
examine this
potential within
#uices, and, if
possible, toprovide
correlates
between mold
numbers and
patulin levels
within #uice.
Fith the
possibility of
fungi actively
producingpatulin within
#uice, control
measures
should aim not
only at the
reduction of
patulin itself,
but the
inhibition of
both fungal
growth and
patulin
productionwithin #uice.
Some studies
have examined
the effects of
many patulin-
reducing
treatments on
live patulin-
producing
fungi. Sulfur
dioxide, sodium
ben?oate, and
potassium
8/20/2019 SIKLUS BIOSINTESIS PATHWAY
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sorbate have
been
investigated for
the affect on B.
nivea growth
and patu-lin
production
within #uice.
Seventy-five
ppm sulfur
dioxide, )6!ppm potassium
sorbate, and
6!! ppm
sodium
ben?oate all
signif-icantly
retarded B.
nivea growth
and patulin
production
$
8/20/2019 SIKLUS BIOSINTESIS PATHWAY
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P. expansum
growth $
8/20/2019 SIKLUS BIOSINTESIS PATHWAY
41/58
;, Atindehou D. &!!).%Vtermination de la Patuline dans le Eus de ruits
;ommercialisVsen ;Wte d0=voire. Sci Aliment &)J)K
&!*.
A(erstrand >, olander A, Andersson A, Cilsson @.
)5*. 7ccurrence of moulds and mycotoxins in fro?en
blueberries. 8Xr Hda &:J)5K&!!.
Andersson AD, Eosefsson @, Cilsson @, A(erstrand >.
)55. Hgelsvampar och Patulin = ru(t 7ch 4Yr. 8Xr
Hda :J&&K:.
Annous 4A, Sapers @, attra??o A, ,
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Proceedings of the &th ycotoxin For(shopB &!!&
Eun 3-*B 4erlin, @ermany.
Arti( C, Acar E, >abraman C, Poyra?oglu D. &!!).
Dffects of various clarification treatments on patulin,
phenolic compound, and organic acid compositions of
apple #uice. Dur ood lein
8/20/2019 SIKLUS BIOSINTESIS PATHWAY
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1;7%DQ2 ;ommittee on ood Additives and
;ontaminants. &!!&. Proposed draft code of practice
for the reduction of patulin contamination in apple #uice
and apple #uice ingredients in other beverages $at step
6 of procedure'. @eneva, Swit?erland. Eoint A7+F/7
ood Standards Programme. p )K5.
;onway FS, Eanisiewic? FE, >lein E%, Sams ;D.
). Strategy for combining heat
treatment, calcium infiltration, and biological control
to reduce postharvest de-
cay of @ala0 apples. /ort Sci 3$'J5!!K.
;ooray iessling >-/, 9indahl->iessling >. ):&.
he effects of patulin and pat-ulin-cysteine mixtures on
%CA synthesis and the freuency of sister-chromatid
exchanges in human lymphocytes. ood ;hem oxicol
&!J:3K:.
;orbett %. &!!3. PatulinRU.>. producers perspective
=nJ Patulin technical sympo-sium. ebruary ):K),
&!!3B >issimmee, la. Cational ;enter for ood safety
adn echnology. Summit, =ll.
%emirci , Arici , @umus . &!!3. Presence of
patulin in fruit and fruit #uices produced in ur(ey.
Drnaehrungs-Umschau 6!$5'J&*&K3.
%e Sylos ;,
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18 ;7P
8/20/2019 SIKLUS BIOSINTESIS PATHWAY
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Patulin control in foods . . .
ran( />, 7rth ,
>ennedy 4P;, Scott
P. )53b.7ccurrence of patulin
and patulin-producing
strains of Penicillium
expansum in natural
rots of apple in
;anada. ;an. =nst.
ood Sci echnol E
*J&&K6.
/arwig E, 4lanchfield
4E, Scott P. )5:.
Patulin production by
Penicillium roque-forti
hom from grape.
;an =nst ood Sci
echnol E ))$3'J)K
6).
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/ate? , @aye .
)5:. =nhibition of
translation in
reticulocyte by the
mycotoxin patulin.
D4S 9ett 6J&6&K*.
/ayes AF, Phillips
%, Filliams F9,
;iegler A. )5.
Acute toxicity of
patulin in mice and
rats. oxicology
)3J)K)!!.
/eatley C@, Philpot
E. )5. he routine
examination for
antibiotics produced
by molds. E @en
icrobiol )J&3&K5.
/offman /, int?laff
/E, Alperden =,
9eistner 9. )5).
Untersuchung uber
die =n-a(tiviering des
y(otoxins Patulin
durch
Suofydrylgruppen.
%ie leis-
chwirtschaft.
6)J)63K*, )63.
/op(ins E. )3. he
toxicological ha?ardsof patulin. 4r =nd 4iol
, Pallaroni
9, A(e ;9, 9em(e
S9, /errera P,
Phillips %.
&!!!. %evelopment
and characteri?ation
of a carbon-based
composite material
for reducing patulin
levels in apple #uice. E
ood Prot *3$)'J)!*K
)!.
=i#imia /, Dbi?u(a G,
San(awa U. ):*.
4iosynthesis ofpatulin, in vitro
conversion of gentisyl
alcohol into patulin by
microsomal
en?yme$s' and
retention of one of the
carbinol protons in
this reaction. ;hem
Pharm 4ull
3$:'J363K5.
=yengar , ;hirtel SE,
er(-
er
8/20/2019 SIKLUS BIOSINTESIS PATHWAY
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#uice. Cahrung+ood
*$)'J3)K3.
>orte A. ):!.
;hromosomal
analysis in bone
marrow cells of
;hinese hamsters
after treatment with
mycotoxins. utat
or?ybs(i ,
>ows?y(-@indifier L,
>urylowic? S. )*5.
Antibiotics origin,
nature, and
properties 8ol. ==.
Cew Gor(J Pergamon
Press. p )&&3K3!.
>ryger yria(ides . &!!).
%iffusion of patulin in
the flesh of pears
inoculated with four
post-harvest
pathogens. E Phy-
topathol
)J65K*).
9am >S, Ceway E7,
@aucher @. )::.
!n vitro
stabili?ation
of *-methylsalicyl-
ic acid
synthetase from
Penicillium urticae.
;an
E icrobiol
3J3!K5.
9arsen 7, risvad
E;, -S,
, Sung F4. )3.
Simultaneous thin
layer
chromatographic
8/20/2019 SIKLUS BIOSINTESIS PATHWAY
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determination of
?earalenone and
patulin in mai?e. E
Planar
;hromatography-
odern 9; *J&5K5.
9indroth S, von
Fright A. )!.
%etoxification of
patulin by adduct
formation with
cysteine. E Dnviron
Pathol oxicol 7ncol
)!$K6'J&6K.
9lovera , 8iladrich
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sum by polymerase
chain reaction. =nt E
ood icrobiol
:J)3K.
ayer 8F, 9egaror
S. )*. Production
of petite mutants of
Saccharomyces cer-
evisiae by patulin. E
Agric ood ;hem
)5J6K*.
c;allum E9, sao
8/20/2019 SIKLUS BIOSINTESIS PATHWAY
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urphy @, 9ynen .
)56. Patulin
biosynthesisJ he
metabolism of m-
hydroxyben-?yl
alcohol and m-
hydroxyben?aldehyde
by particulate
preparations from
Peni-cillium patulum.
Dur E 4iochem
6:J*5K56.
utlu , /i?arcioglu
C, @o(men 8. )5.Patulin adsorption
(inetics on activated
carbon, activation
energy, and heat of
adsorption. E ood
Sci *&J)&:K3!.
Ceway E, @aucher
@. ):). =ntrinsic
limitations on the
continued production
of the antibiotic
patulin by Penicillium
urticae. ;an E
icrobiol &5J&!*K)6.
Corstadt A, c;alla
. )*3. Phytotoxic
substance from a
species of Penicilli-
um. Science
)!J)!K).
Corstadt A, c;alla
. )*:.
icrobiological
population in
stubblefield mulched
soil. Soil Sci )!5J)::.
Cortholt %, 8an
Dgmond /P, Paulsch
FD. )5. Patulin
production by some
fun-gal species in
relation to water
activity and
temperature. E ood
Prot )$))'J::6K !.
7ugh ;S, ;orison
;A. ):!.
easurement of
patulin in grapes and
wines. E ood Sci
6J5*K:.
7swald /, ran( />,
>omitows(i %, Finter
/. )5:. 9ong-term
testing of patulin
administered orally to
Sprague-%awley rats
and Swiss mice.
ood ;osmet oxi-col
)*J&3K5.
Par( %9, 9ee 9S,
Price
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;
8/20/2019 SIKLUS BIOSINTESIS PATHWAY
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@;+S confirmation of patulin identity. =nJ ruc(sess
and others, editors. ycotoxins and food safety. Cew
Gor(J >luwer Academic+Plenum Pub-lishers. p )36K!.
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Singh E. )*5. Patulin. =nJ @ottlieb %,
Shaw P%, editors. Antibiotics =J mechanics
of action. 4erlinJ Springer-8erlag. p
*&)K3!.
Sommer C, 4uchanan E
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httpJ++www.usda.gov+nass+pubs+agr!&+!&[ch6.pdf .
Accessed
arch 6, &!!3.
1US%A2 U.S. ood and %rug Administration. &!!.
;ompliance policy guide. ;om-pliance policy guidance
for %A staff. Sec. 6)!.)6!. Apple #uice, apple #uice
concentrates,
and apple #uice productsRAdulteration with patulin.
Available
fromJ
httpJ++www.fda.gov+ora+compliance[ref+cpg+cpgfod+cpg
6)!-)6!.htm.
Ac-
cessed
arch
&, &!!.
8alletrisco S, ;asadio S, Stefanelli ;. )!. Use of
ultraviolet radiation to brea( down mycotoxins.
=ndustrie Alimentari &$&::'J))))K&.
van Dgmond /P. ):. ;urrent situation on
regulations for mycotoxins. 7verview of tolerances
and status of standard methods of sampling and
analysis. ood Addit ;ontam *J)3:K::.
8an 9ui#( A. )3:. Antagonism of Penicillium spp.
versus Pythium de"aryanum. ;hron 4ot J&)!.
Fal(er E, Fiesner 4P. ). Patulin and clavicin.
9ancet &*J&.
Fang =>, oehler PD. ):5. Presence
of patulin in pasteuri?ed apple cider. E ood Sci
6&J5K:!.
Fichmann @, /erbarth 7, 9ehmann =. &!!&. he
mycotoxins citrinin, gliotoxin, and patulin affect
interferon- rather than interleu(in- production in
human blood cells. Dnviron oxicol )5$3'J&))K:.
Fisniews(y A, @lat? 4A, @leason 9,
8/20/2019 SIKLUS BIOSINTESIS PATHWAY
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Patulin control in foods . . .
ood Prot *3J5!3K:.
1F/72 Forld /ealth
7rgani?ation. )!.
Dvaluation of certain
food additives and
contaminants. F/7
thirty-fifth report of
the #oint A7+F/7
expert committee on
food additives.
@enevaJ echnical
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Vol. 1, 2005 — ;7P