PATULIN
1. EXPLANATION
Patulin has not been previously evaluated by the Joint FAO/WHO
Expert Committee on Food Additives.
Patulin is a mycotoxin produced by fungi belonging to several
genera including Penicillium, Aspergillus and Byssochlamys.
Although patulin can occur in many molding fruits, grains and other
foods, the major source of patulin contamination is in apples with
brown rot, and in apple cider or apple juice. Patulin is stable in an
acid environment, and is not destroyed during thermal processing
(Pohland & Allen, 1970; Scott & Somers, 1968). Patulin toxicosis has
been described in cattle and poultry (Camguilhem et al., 1976;
Schultz et al., 1969; Lovett, 1972).
Many of the older studies on patulin levels in fruit and fruit
products employed methodology that lacked sufficient sensitivity. In
addition, positive identification of patulin has not always been
confirmed. Based on the results of more recent surveys conducted in
several countries, the mean patulin content of apple juice/beverage is
estimated to be less than 10-15 µg/l (based on the values of the
positive samples and assuming that patulin is at the detection limit
in the non-detected samples) with an overall range of less than 1 to
250 µg/l patulin and a contamination range of 7-52% in the retail
product. (Mortimer et al., 1985; Anderson & Josephson, 1979). The
dietary intake of patulin from apple juice containing 10-15 µg/l is
estimated to range from less than 0.03 to 0.26 µg/kg bw/day (less than
l.9-3.9 µg/day) for different age groups in the population, including
children. Because of the limited number of countries upon which these
estimates are based they cannot automatically be applied worldwide.
Patulin has been reviewed by IARC (1976 & 1986).
2. BIOLOGICAL DATA
2.1 Biochemical aspects
2.1.1 Absorption, distribution and excretion
A single oral dose of 3 mg/kg bw of [14C] (0.133 mCi/mmol)
patulin in 1mM citrate buffer was given to 17 male and 12 female
Sprague Dawley rats that had been exposed for 41-66 weeks after birth
to levels of 0 or 1.5 mg/kg bw of patulin in 1 mM citrate buffer. All
animals were fasted for 24 hours before the administration of labelled
patulin. Animals were placed in metabolic cages and feces, urine and
CO2 were collected. One or two animals of each sex from each group
(untreated or pretreated with patulin) were sacrificed at 4, 24, 48,
or 72 hours or 7 days after blood was collected. Concentrations of
patulin in erythrocytes were calculated from the difference between
radioactivity of whole blood and serum. Within 7 days approximately
49% of administered radioactivity was recovered from feces, and 36%
from urine. Most of the excretion of label occurred within the first
24 hours. Approximately 1-2% of the label was recovered as 14CO2.
At the end of 7 days, 2-3% of the radioactivity was recovered in soft
tissues and blood. The major retention sites of patulin were
erythrocytes and blood-rich organs (spleen, kidney, lung and liver)
(Dailey et al., 1977a).
2.1.2 Effects on enzymes and other biochemical parameters
Oxygen uptake stimulated by Krebs-cycle intermediates was
reported in an abstract to be inhibited in tissue extracts from mice,
rat and golden hamster. Inhibition of oxygen uptake in liver
homogenates was observed at levels of patulin as low as 0.033 mM.
Inhibition of oxygen uptake in heart and muscle homogenates was
greater than in liver homogenates. Patulin competitively inhibited
succinate dehydrogenase in mouse liver homogenates. The P/O ratio was
not affected by the toxin. In comparative studies, the golden hamster
was more susceptible, and the rat less susceptible to patulin
inhibition than the mouse (Hayes, 1977).
Kidney explants from male Osborne-Mendel rats, when incubated for
18 hours in medium containing 0.5, 0.75, or 1.0 mM of patulin
in vitro, lost their respiratory ability as measured by conversion
of [14C] glucose to 14 CO2. During measurement of respiration,
patulin was not present in the reaction mixture. At 0.1 mM of
patulin, respiration was increased. Leakage of protein into the
medium at a concentration of 1.0 mM patulin may indicate increased
cell membrane permeability (Braunberg et al., 1982).
Absorption of radiolabelled glycine, alanine and lysine was
reduced in perfused intestines of rats that had received 100 µg of
patulin intraperitoneally on alternate days for 1 month (dose
aproximately 1.6 mg/kg bw/day). The authors attributed this effect to
reduced total ATPase, NaK ATPase, and alkaline phosphatase activities
which were studied in a satellite group of rats (Devaraj et al.,
1982a).
Forty-eight hours after i.p. injection of 5.0 or 7.5 mg/kg bw of
patulin into male ICR mice, NaKATPase and MgATPase of liver, kidney
and brain preparations were significantly inhibited. Injection of 2.5
mg/kg bw of patulin had no significant effect on enzyme activity. The
same effects were demonstrated in in vitro studies with
mitochondrial and microsomal fractions of liver, kidney and brain of
ICR mice (Phillips & Hayes, 1977).
Patulin inhibited the in vitro activity of NaKATPase in
microsomes prepared from mouse brain. Activity was partially restored
by washing. Preincubation of patulin with dithiothreitol or
glutathione prevented the inhibition (Phillips & Hayes, 1978).
Patulin inhibited acetylcholinesterase and NaKATPase in cerebral
hemisphere, cerebellum and medulla oblongata in rats treated for 1
month with approximately 1.6 mg/kg bw/day of patulin injected
intraperitoneally. Concomitantly, acetylcholine levels were raised in
these brain segments (Devaraj et al., 1982b).
A non-competitive and irreversible inhibition of the activity of
alcohol dehydrogenase derived from yeast was attributed by the authors
to patulin's ability to bind to SH-groups; the Ki was found to be
5.0 x 10-5 M (Ashoor & Chu, 1973a).
Non-competitive inhibition was demonstrated when patulin was
inclubated with rabbit-muscle aldolase; the ki was 1.3 x 10-5 M. The
cysteine adduct of patulin was a less effective inhibitor (Ashoor &
Chu, 1973b).
Patulin, at a level of 4.348 µmol/ml, was reported to inhibit
in vitrothe activity of DNA-dependent RNA polymerases I and II
prepared from rat liver nuclei by 29% and 84%, respectively (Tashiro
et al., 1979).
Patulin at a level of 200 µg/ml inhibited in vitro the chain
initiation stage of RNA synthesis in rat liver nuclei (Moule & Hatey,
1977).
Inhibition of yeast-derived aminoacyl-tRNA synthetase by patulin
occurs mainly by modification of sulfhydryl groups on the enzyme
(Arafat, et al., 1985).
Ribonuclease H, prepared from rat liver nuclei, was inhibited by
patulin in vitro by 62% at a concentration of 0.324 µmol/mol, and by
47% at a concentration of 1.071 µmol/ml (Tashiro et al., 1979).
Acid RNAse in human placental microsome and mitochondria-rich
fractions was increased up to 1.5 times when incubated with 0.5 - 3
mg/g placenta of patulin in vitro (Fuks-Holmberg, 1980).
Patulin caused a competitive inhibition of lactate dehydrogenase
from rabbit muscle (Ki = 7.2 x 10-5 M). The presence of cysteine
reversed the inhibitory effect of patulin on lactate dehydrogenase
(Ashoor & Chu, 1973a).
Liver lactate dehydrogenase was increased in 4 pregnant Sprague
Dawley rats after exposure to 3 mg/kg bw/day of patulin in tris-
acetate buffer, by gavage, from days 1-19 of gestation (Fuks-Holmberg,
1980).
Malate dehydrogenase in human placental microsome- and
mitochondria-rich fractions was increased up to 15 times when
incubated with 0.5 - 3 mg/g placenta of patulin in vitro
(Fuks-Holmberg, 1980).
Placental GPT was depressed in 4 pregnant Sprague Dawley rats
after exposure to 3 mg/kg bw/day of patulin in tris-acetate buffer, by
gavage, from days 1-19 of gestation (Fuks-Holmberg, 1980).
When white male albino mice were injected with 10 doses of 0.1 mg
of patulin in propylene glycol on alternate days, glycogen
phosphorylase in the liver was activated, and blood glucose levels
increased by 60%. These results were confirmed by studies in vitro
(Madiyalakan & Shanmugasundaram, 1978).
Groups of 10 rats were fed either regular diet, diet infected
with Penicillium patulum, or were injected intraperitoneally with
purified patulin (1 mg/kg bw on alternate days) for 3 months. Fasting
blood glucose levels were elevated and a glucose tolerance test
revealed an elevated glucose curve and reduced insulin production. The
authors concluded that patulin is diabetogenic (Devaraj et al.,
1986).
Four days after a single intraperitoneal dose of 0, 0.5, 5.0, or
10.0 mg/kg bw of patulin to male Fischer 344 rats, liver mixed
function oxidases and cytochrome P450 activity were determined.
Oxidative cleavage of phosphonothioate EPN and aryl hydrocarbon
hydroxylase were elevated at 10 mg/kg bw. No effect was observed on
p-nitroanisole O-demethylase or on cytochrome P450 (Kangsadalampai
et al., 1981).
Patulin was reported in an abstract to induce mixed function
oxidase in male ICR mice treated with 0.5, 1.0 or 2.0 mg/kg bw of
patulin intraperitoneally (Siraj & Hayes, 1978).
2.2 Toxicological studies
2.2.1 Acute toxicity
LD50
Species Sex Route (mg/kg bw) Reference
Mouse M oral 29-48 Escoula et al., 1977
Lindroth & von Wright, 1978
McKinley & Carlton, 1980a
F 46.31 Escoula et al., 1977
M&F 17 Hayes et al., 1979
? 25 Katzman et al., 1944
M i.p. 5.7-8.17 Ciegler et al., 1976
Escoula et al., 1977
McKinley & Carlton, 1980a
F 10.85 Escoula et al., 1977
M&F 7.6 Hayes et al., 1979
? 4-5.7 Katzman et al., 1944
Ciegler et al., 1976
M i.v. 8.57 Escoula et al., 1977
F 8.57 Escoula et al., 1977
? s.c. 8-10 Katzman et al., 1944
M 10 McKinley & Carlton, 1980a
Rat M oral 30.53-55.0 Escoula et al., 1977
McKinley et al., 1982
F 27.79 Escoula et al., 1977
? 32.5 Dailey et al., 1977b
M&F 108-118 Hayes et al., 1979
LD50
Species Sex Route (mg/kg bw) Reference
M i.p. 4.59-10.0 Escoula et al., 1977
McKinley et al., 1982
(neonate) F 5.70 Escoula et al., 1977
M&F 6.8 Hayes et al., 1979
(weanling) M&F 5.9 Hayes et al., 1979
M i.v. 8.57 Escoula et al., 1977
M s.c. 11.0 McKinley et al., 1982
? 25 Katzman et al., 1944
Hamster M oral 31.5 McKinley & Carlton, 1980b
i.p. 10 McKinley & Carlton, 1980b
s.c. 23 McKinley & Carlton, 1980b
Acute toxicity of i.p.-administered patulin was reported to be
reduced by simultaneous administration of another mycotoxin,
rubratoxin B (Kangsadalampai et al., 1981). Toxic signs
consistently reported in all studies were agitation, in some cases
convulsions, dyspnea, pulmonary congestion and edema, and ulcerations,
hyperemia and distension of the gastro-intestinal tract. When a
patulin/cysteine adduct was administered to mice intraperitoneally, no
acute toxicity was observed at levels up to 150 µg of patulin/mouse
(Ciegler et al., 1976).
2.2.2 Short-term Studies
2.2.2.1 Mice
When patulin was administered by gavage in citrate buffer to
groups of 10 male Swiss ICR mice at doses of 0, 24 or 36 mg/kg bw,
daily or on alternate days for 14 days, body weight was depressed and
mortality was increased in a dose dependent manner. Histopathological
lesions were found in the gastro-intestinal tract, which included
epithelial degeneration, hemorrhage, and ulceration of gastric mucosa,
and exudation and epithelial desquamation in the duodenum (McKinley &
Carlton, 1980a).
2.2.2.2 Rats
When patulin was administered by gavage to groups of 10 male
Sprague Dawley rats at doses of 28 or 41 mg/kg bw, daily or on
alternate days for 14 days, initial loss of body weight was observed;
animals recovered after day 4. Mortality was increased in all treated
groups, but no dose dependency was observed. Gross lesions were found
in the stomach and small intestine; the gastric mucosa was reddened
and the stomach was distended. The duodenum and jejunum were distended
by fluid. Histopathological lesions were found in the stomach which
consisted of ulceration of the mucosa, epithelial degeneration,
hemorrhage, and neutrophil and mononuclear cell infiltration (McKinley
et al., 1982).
Drinking water containing 0, 24, 84, or 295 mg/l of patulin in
1 mM citrate buffer was given to groups of 6 SPF RIVM:Tox (Wistar
derived) rats for 4 weeks. Food and liquid intake were recorded three
times per week. Body weights were determined at the start of the
experiment and at termination. Urinalysis, including urine volume,
bilirubin, and urinary protein were determined in the last week.
Creatinine clearance was calculated from serum and urine levels of
creatinine. At termination, the animals were examined
macroscopically, and the liver, spleen, thyroid glands, brain,
kidneys, heart, mesenteric lymph nodes, adrenal glands, thymus, testes
and ovaries were weighed. Histopathological examination was carried
out on all organs and tissues of the high-dose and the control groups.
Food and liquid intake were reduced in the mid- and high-dose animals.
Body weights at the high dose level were decreased. Creatinine
clearance was lower in the high-dose animals, but no morphological
glomerular damage was observed. In the high-dose group, fundic ulcers
in the stomach were observed in combination with enlarged and active
pancreatico-duodenal lymph nodes, while villous hyperemia of the
duodenum was observed at the mid- and high-dose levels. The authors
suggested, based on normal appearance of the adrenal glands, that the
observed effects in the gastrointestinal tract were a direct effect of
patulin on the tissue, which was not mediated through adrenal gland
stimulation (stress) (Speijers et al., 1988).
2.2.2.3 Hamsters
When patulin was administered by gavage to groups of 10 male
Syrian golden hamsters at doses of 0, 16 or 24 mg/kg bw, daily or on
alternate days for 14 days, loss of body weight was observed and
mortality was increased in all treated groups, but no dose dependency
was observed. Gross lesions were found in the stomach and duodenum.
Histopathological lesions were found in the gastro-intestinal tract
that included epithelial degeneration, hemorrhage and ulceration
(McKinley & Carlton, 1980b).
2.2.2.4 Monkeys
Groups of 1 male and 1 female pigtail monkeys (Macaca nemestrina)
received daily doses of 0, 5, 50, or 500 µg/kg bw/day of patulin for
4 weeks. Monkeys of the highest dose group received 5 mg/kg bw/day
patulin for 2 additional weeks. Weekly determinations were made of
SGOT, SAP, BUN, cholesterol, sodium and potassium as well as
hematological parameters. Plasma protein electrophoresis was
performed and glucose and lipoprotein levels were determined. No signs
of toxicity were noted, except that the monkeys receiving 5 mg/kg
bw/day of patulin started to reject their food during the last 3 days
of the experiment. No statistically significant differences were
observed in any of the parameters studied (Garza et al., 1977).
2.2.3 Long-term/carcinogenicity studies
2.2.3.1 Mice
Twelve pregnant Swiss mice received 2 mg/kg bw/day of patulin in
water containing 0.05% lactic acid twice daily by gavage for 6 days
starting 14 days after mating. The 12 control mice received 0.05%
lactic acid by gavage. Mean survival time was significantly reduced
in the patulin-treated dams, while 2 of 12 control animals and 5 of 12
experimental animals developed tumours. Of the offspring, 8 of 43 male
and 11 of 52 female suckling mice died in the first 6 days of life,
with hyperemia and bleeding in the brain, lungs and skin. When these
early deaths were excluded from the calculations, patulin did not
affect survival time in the animals exposed in utero. No evidence of
carcinogenicity was observed in the offspring that had been exposed
only to patulin in utero, (Osswald et al., 1978).
2.2.3.2 Rats
Subcutaneous injections of 0.2 mg of patulin in 0.5 ml of arachis
oil administered biweekly for 61 or 64 weeks to 2 groups of 5 male
Wistar rats weighing 100 g at the start of the experiment was reported
to produce local (fibro)sarcomas at the injection site in 4 of 4 and
2 of 4 rats surviving at the time when the first tumour was observed.
No metastases were observed, and of 3 tumours tested, only one was
transplantable in 3 of 12 recipient rats. Control animals receiving
arachis oil did not develop local tumours (Dickens & Jones, 1961).
When patulin in water containing 0.05% lactic acid, was
administered by gavage twice weekly to 50 female SPF Sprague Dawley
rats at a dose of 1 mg/kg bw for 4 weeks, and 2.5 mg/kg bw for the
following 70 weeks (total dose: 358 mg/kg bw of patulin), no effects
were observed on weight gain or on survival. No significant
differences were observed in tumour incidence. The occurrence of 4
forestomach papillomas and 2 glandular stomach adenomas, as compared
to none in the control animals, is noteworthy. The Committee noted a
discrepancy between the reported duration of the study (64 weeks) and
the reported duration of administration (74 weeks) (Osswald et al.,
1978).
Groups of 70 FDRL Wistar rats of each sex were exposed to 0, 0.1,
0.5, and 1.5 mg/kg bw/day of patulin in citrate buffer by gavage 3
times per week for 24 months. The rats were derived from the F1
generation of a 1-generation reproduction study. Mortality was
increased in both sexes in the highest dose: all males had died by 19
months; 19% of females survived until termination at 24 months. Body
weights of males were reduced in the mid- and high-dose, but for
females body weights were comparable in all groups. No difference in
tumour incidence was observed (Becci et al., 1981).
2.2.4 Reproduction studies
2.2.4.1 Rats
Groups of 30 Sprague-Dawley rats of each sex received doses of 0,
1.5, 7.5, and 15.0 mg/kg bw/day of patulin in citrate buffer by gavage
5 times per week for 7 weeks before mating. The pregnant dams were
gavaged daily at the same levels during gestation. Half the dams were
sacrificed on day 20 of gestation, and used for teratological
evaluation. The remaining dams were allowed to produce the F1
generation. Some of the F0 and F1 males were used for a dominant
lethal experiment. Twenty-three controls and 15 low-dose animals per
sex were continued to produce an F2 generation. One-half of the
latter generation were again used for teratological evaluation.
Hematology and blood chemistry examinations were performed on 10 males
and 10 females of the F1 generation 23 days after weaning. The only
lesion found at necropsy of parent animals was gaseous distension of
the gastrointestinal tract. All treated males of the F0 generation
had a dose-related reduction in weight gain. High mortality occurred
at 7.5 and 15.0 mg/kg bw/day in both males and dams. Although litter
size at 7.5 mg/kg bw/day was comparable to controls, survival of male
progeny was severely impaired. At the 1.5 mg/kg bw/day level, pup
growth of both sexes was reduced, and there was increased mortality
among the F2 females. No significant alterations were found in the
hematology and blood chemistry levels in selected animals of the F1
generation (Dailey et al., 1977b).
Groups of 50 FDRL Wistar rats of each sex were exposed to 0, 0.1,
0.5, and 1.5 mg/kg bw/day of patulin in citrate buffer by gavage for
4 weeks before mating, and pregnant females were dosed through
gestation and lactation. The parent generation was sacrificed after
weaning. Body weight gain was comparable among groups. In the high
dose group, 10 females died. Reproductive parameters such as mating
success, litter size, fertility, gestation, viability, and lactation
indices, and pup weight at birth, 4 days and at weaning, were not
statistically different among experimental groups. Histopathological
evaluation of grossly abnormal tissues of the F0 generation did not
show any effects of patulin treatment. The F1 generation was used for
a 2-year toxicity/carcinogenicity study (see section 2.2.3.2) (Becci
et al., 1981).
2.2.5 Special studies on genotoxicity
Results of genotoxicity assays on Patulin
Concentration
Test System Test Object of Patulin Results Reference
DNA synthesis T.pyriformis 3.2 µg/ml Positive Burger et al.
retardation 1988
DNA synthesis AWRF cells 1,2,4&8 µg/ml Positive Stetina &
retardation CHO cells 0.25, 0.5, l,2,4 Votava, 1986
µg/ml
DNA breakage ColE1 plasmid 0.25, 0.5, 1.0 Negative Lee &
in vitro DNA & 5.0 mM (1) Roeschenthaler,
Lambda DNA 0.5, 1, 5, 10 1986
& 14 mM
DNA breakage E.coli 10, 20, 25 & 50 Positive Lee &
in vivo D110 polA µg/ml Roeschenthaler,
1986
DNA breakage FM3A mouse 1.0, 3.2, 10 Positive Umeda et al.,
mammary carcinoma µg/ml 1977
cells
DNA breakage AWRF cells 2 & 10 µg/ml Positive Stetina &
CHO cells 2, 8 & 10 Votava, 1986
µg/ml
Concentration
Test System Test Object of Patulin Results Reference
Prophage E. coli 5, 10, 25 & 50 Positive Lee &
induction X8011(lambda) µg/ml Roeschenthaler,
1986
Spot test E. coli 1 - 10 µg/ Positive Auffray &
K12 assay (2) Boutibonnes,
1986
Chromotest (3) E. coli 0.01, 0.02 & Positive Auffray &
K12 PQ37 0.05 µg/ml (No S-9) Boutibonnes,
Negative 1987
(with S-9)
Chromotest (2) E. coli 0.001 - 30 Negative Krivobok et
PQ37 µg/ml al., 1987
Recombinogenesis B. subtilis 20 & 100 µg/ Positive Ueno &
H17/M45 disc Kubota, 1976
Ames test E. coli 1 µg/ml (to Positive Burger et
M13am6H1 phage) &/or al., 1988.
phage 5 µg/ml (to
bacteria)
Ames test (3,4) S.typhimurium 0.01, 0.1, 1, Negative Ueno et al.,
TA-98 10, 100 & 500 1978
TA-100 µg/plate
Concentration
Test System Test Object of Patulin Results Reference
Ames test (3) S.typhimurium 0.25, 2.5, 25 & Negative Wehner et
TA-98 250 µg/plate al., 1978
TA-100
TA-1535
TA-1537
Ames test (3) S.typhimurium 0.1, 1, 10 & 100 Negative Kuczuk et
TA-1535 µg/plate al., 1978
TA-1537
TA-1538
Ames test (5) S.typhimurium 5, 10, 20 & 30 Negative von Wright &
TA-98 µg/plate Lindroth, 1978
TA-100
Ames test (3,5) S.typhimurium <0.0065 umoles/ Negative Bartsch et
TA-100 plate al., 1980
TA-1538
Host mediated S.typhimurium 3x <500 µg Negative Gabridge &
assay in G46 i.m. Legator, 1969
Swiss albino
mice
Host mediated S.typhimurium 10 & 20 mg/kg Negative von Wright &
assay in TA-1950 bw, gavage Lindroth, 1978
male NMRI mice TA-1951
Reverse S. cerevisiae 50 (No S-9) & Negative Kuczuk et
mutagenesis (3) D-3 100 (with S-9) al., 1978
µg/plate
Concentration
Test System Test Object of Patulin Results Reference
Forward S. cerevisiae 10, 25, 50 & 75 Positive Mayer &
mutagenesis (haploid) µg/ml Legator, 1969
Forward FM3A mouse 0.032, 0.1 & Positive Umeda et al.,
mutagenesis mammary carcinoma 0.32 µg/ml 1977
(8-azoquanine cells
resistance)
Unscheduled Primary ACI rat 60 & 600 µM Negative Mori et al.,
DNA synthesis hepatocytes 1984
induction in vitro
Primary C3H 65 & 650 µM Negative Mori et al.,
mouse hepatocytes 1984
in vitro
Chromosome FM3A mouse 0.032, 0.1 & Positive Mori et al.,
aberration mammary carcinoma 0.32 µg/ml 1984
induction cells
Chromosome Chinese hamster 1, 2.5, 5 & 10 Positive Thust et al.,
aberration V79-E cells µM (no S-9) 1982
induction (3) in vitro Negative
(with S-9)
Chromosome Human 3.5 µM Positive Withers,
aberration leucocytes 1966
induction in vitro
Concentration
Test System Test Object of Patulin Results Reference
Sister chromatid Chinese hamster 1, 2.5, 5 & 10 Negative Thust et al.,
exchange V79-E cells µM 1982
induction (3) in vitro
Sister chromatid Primary Chinese 0.5,1 & 2 Positive Kubiak &
exchange hamster cells µg/ml Kosz-Vnencha k,
induction in vitro 1983
Sister chromatid Human peripheral 0.075, 0.10, Weakly Cooray et
exchange blood 0.20 & 0.30 positive al., 1982
induction lymphocytes µg/ml at 0.10 &
in vitro 0.20 µg/ml
Cell cycle Primary Chinese 0.5, 1 & 2 Positive Kubiak &
retardation hamster cells µg/ml Kosz-Vnenchak,
in vitro 1983
Cell cycle Human peripheral 0.075 & 0.30 Positive Cooray et
retardation blood lymphocytes µg/ml at 0.30 al., 1982
in vitro µg/ml
Chromosome Chinese hamster 2 x 20 mg/kg Positive Korte et
aberration bone marrow bw gavage (6) al., 1979
induction cells in vivo
Chromosome Chinese hamster 2 x 1, 10 & Positive Korte, 1980
aberration bone marrow 20 mg/kg bw >10 mg/kg
induction cells in vivo gavage
Concentration
Test System Test Object of Patulin Results Reference
Chromosome Chinese hamster 2 x 10 & 20 Positive Korte &
aberration bone marrow mg/kg bw dose Ruckert, 1980
induction cells in vivo response
Sister chromatid Chinese hamster 2 x 1, 10 & Negative Korte, 1980
exchange bone marrow 20 mg/kg bw
induction cells in vivo gavage
Cell cycle Human peripheral 0.075 & 0.30 Positive Cooray et
retardation blood lymphocytes µg/ml al., 1982
in vitro
Dominant ICR/Ha Swiss 0.1 & 0.3 mg/kg Negative Epstein et
lethal assay Mice bw, i.p. al., 1972
Dominant Sprague-Dawley 1.5 mg/kg bw Negative Dailey et
lethal assay rats 5x/wk x 10-11 al., 1977b
wk gavage
Dominant Texas ICR x 3.0 mg/kg bw, Negative Reddy et
lethal assay Sprague Dawley i.p. al., 1978
Sch:Ars(CF1)f
(1) Positive when CuCl2 & NADPH were added
(2) Both with and without S-9 fraction (source not specified)
(3) Both with and without rat liver S-9 fraction
(4) Both with regular plate and preincubation methods
(5) Both with and without mouse liver S-9 fraction
(6) Effect negated if animals first given ethanol as only liquid for 9 wk prior to exposure
2.2.6 Special studies on neurotoxicity
2.2.6.1 Rats
Groups of 25 albino rats (sex not specified) weighing 25 - 30 g
received 0 or 100 µg of patulin in propylene glycol intraperitoneally
on alternate days (dose approximately 1.6 mg/kg bw/day) for 1 month.
The patulin treated animals showed convulsions, tremors, impaired
locomotion, stiffness of hindlimbs, and wagging of the head. Patulin
inhibited acetylcholinesterase and NaKATPase in the cerebral
hemisphere, cerebellum and medulla oblongata. Concomitantly,
acetylcholine levels were raised in these brain segments (Devaraj
et al., 1982a).
2.2.7 Special studies on teratogenicity
2.2.7.1 Rats
As part of a 2-generation reproduction study (see section
2.2.4.1), offspring of 15 Sprague Dawley dams of the F1 and F2
generation exposed by gavage to 0 or 1.5 mg/kg bw/day of patulin in
citrate buffer were evaluated for teratological abnormalities. Patulin
caused an increase in resorptions in the F1 litters, but this effect
was not observed in the F2 generation. The average weight of male
fetuses of the F2 generation was significantly less than controls. No
increases in skeletal or soft tissue abnormalities were observed
(Dailey et al., 1977b).
However, when patulin was administered i.p. to groups of 10 - 17
pregnant Charles River CD1 rats at doses of 1.5 or 2.0 mg/kg bw/day,
a significant decrease in average fetal body weight was observed at
the lower dose, and at 2.0 mg/kg bw/day all implanted embryos were
resorbed (Reddy, et al., 1978).
Patulin was injected into the air cell of chick eggs. It was
reported to be embryotoxic at levels of 2.35-68.7 µg/egg depending on
the age of the embryo, and teratogenic at levels of 1-2 µg/egg.
Patulin/cysteine adducts exhibited the same toxic effects, but at much
higher doses: 15-150 µg of patulin equivalents (Ciegler et al.,
1976).
2.3 Observations in man
Patulin has been tested as an antibiotic for treatment of the
common cold in humans. Application was through the nasal route
(1:10,000 or 1:20,000 solutions, every 4 hours). Most of the
information is anecdotal (Gye, 1943). A report on a controlled trial
failed to identify the number of patients tested, and was unclear as
to which clinical tests were performed to support the author's
assertion that no ill effects were observed (Hopkins, 1943).
3. COMMENTS
The Commitee reviewed studies on the biochemistry and toxicology
of patulin as well as very limited information pertaining to
observations in humans when patulin was tested as an antibiotic for
treatment of the common cold in humans.
In rats, most of the administered dose was eliminated within 48
hours in feces and urine. Less than 2% was expired as CO2. No other
metabolites have been identified. About 2% of the administered dose
was present after 7 days, primarily associated with erythrocytes.
Patulin has a strong affinity to sulfhydryl groups. Patulin
adducts formed with cysteine are less toxic than the unmodified
compound in acute toxicity, teratogenicity, and mutagenicity studies.
Its affinity for SH-groups explains its inhibitory activity on many
enzymes.
In acute and short-term studies, patulin caused gastrointestinal
hyperemia, distension, hemorrhage and ulceration. Pigtail monkeys
tolerated patulin consumption of up to 0.5 mg per kg of body weight
per day for 4 weeks without adverse effects.
Two reproduction studies in rats were available. No reproductive
or teratogenic effects were noted at levels of up to 1.5 mg/kg of
bw/day, but there was an increase of resorptions at that level.
An oral carcinogenicity study in rats was negative. Short-term
in vitro genotoxicity studies indicate that patulin is not
mutagenic, but that it has clastogenic activity in some test systems.
A provisional tolerable weekly intake (PTWI) for patulin of 7
µg/kg bw was set based on a no-effect level of 0.1 mg/kg bw in a
combined reproduction/long-term/carcinogenicity study. An additional
long-term carcinogenicity study in a rodent species other than the rat
is recommended for further evaluation of the toxicity of patulin.
Data on patulin levels in apple juice, a food that is often
consumed by children, were available. In this group of the population
and based on surveys in limited areas of the world a maximum intake of
0.26 µg/kg bw/day has been estimated. However, occasional samples of
apple juice can be heavily contaminated and therefore efforts should
be made to avoid unnecessary exposure to this mycotoxin by adherence
to good manufacturing practices in which rotted or mouldy fruit are
not used. This should reduce dietary exposure to levels below the
PTWI. The Committee urged the application of such practices.
4. EVALUATION
Level causing no toxicological effect
Rat: 0.1 mg/kg bw/day
Estimate of Provisional Tolerable Weekly Intake
7 µg/kg bw
Further work or information desirable
A long-term/carcinogenicity study in a rodent species other than
the rat.
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See Also:
Toxicological Abbreviations
Patulin (WHO Food Additives Series 35)
PATULIN (JECFA Evaluation)
Patulin (IARC Summary & Evaluation, Volume 10, 1976)
Patulin (IARC Summary & Evaluation, Volume 40, 1986)