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Toxicological information

Toxicity to reproduction

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Administrative data

Endpoint:
two-generation reproductive toxicity
Remarks:
based on test type (migrated information)
Type of information:
read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study (GLP)
Justification for type of information:
The extrapolation from sorbic acid to potassium sorbate or vice versa is considered not to be restricted in any way, since the determinant of potential toxicity is on the "sorbate" anion.

Data source

Referenceopen allclose all

Reference Type:
study report
Title:
Unnamed
Year:
2004
Report Date:
2004
Reference Type:
study report
Title:
Unnamed
Year:
2004
Report Date:
2004

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 416 (Two-Generation Reproduction Toxicity Study)
Version / remarks:
adopted 2001-01-22
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.35 (Two-Generation Reproduction Toxicity Test)
Version / remarks:
, 1988
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
signed 2003-12-08
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material: Sorbic acid
- Physical state: White crystalline powder
- Batch No.: 0000009738
- Purity: 99.9%
- Expiration date of the lot/batch: March 27, 2006
- Stability: The test substance was considered stable for the duration of the study

Test animals

Species:
rat
Strain:
Crj: CD(SD)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland GmbH, Sulzfeld, Germany
- Age at study initiation:  55-61 days
- Weight at study initiation: Males: 167.3-253.2 g Females: 149.1-187.7 g

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
other: 0.5 % aqueous hydroxypropyl-methylcellulose gel
Details on exposure:
VEHICLE
- Concentration in vehicle: 0, 30, 100 and 300 mg/mL

TOTAL VOLUME APPLIED: 10 mL/kg b.w./day


Details on mating procedure:
Mating: One male with one female from the same dose level. The day on which sperm or a vaginal plug was found was considered as the day of conception (day 0 of pregnancy).
Duration of mating: up to 3 weeks
Deviations from standard protocol: None (no standardisation of litter sizes was carried out, and no second matings were conducted)
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Recovery was invariably better than 80 % of nominal, indicating that doses were close to nominal and homogeneousyl distributed in the matrix.
Duration of treatment / exposure:
- Duration of exposure before mating: 10 weeks
- Duration of exposure in general:
Males (P-, F1-generation): until termination of mating period
Females (P-, F1-generation): until day 21 of lactation (weaning)
F1-pups not selected for mating: until 7 weeks after birth
F2-generation, males and females: until day 21 of lactation.
Frequency of treatment:
Daily
Details on study schedule:
Number of generation studies: 2
Doses / concentrations
Remarks:
Doses / Concentrations:
0, 300, 1000, 3000 mg/kg bw/day
Basis:

No. of animals per sex per dose:
Number of animals per group:
Control group (P mating): 30 males / 30 females
300 mg/kg/day (P mating): 30 males / 30 females
1000 mg/kg/day (P mating): 30 males / 30 females
3000 mg/kg/day (P mating): 30 males / 30 females
Control group (F1 mating): 25 males / 25 females
300 mg/kg/day (F1 mating): 25 males / 25 females
1000 mg/kg/day (F1 mating): 25 males / 25 females
3000 mg/kg/day (F1 mating): 25 males / 25 females
Control animals:
yes, concurrent vehicle

Examinations

Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Cage side observations checked: Clinical signs

BODY WEIGHT: Yes

FOOD CONSUMPTION: Yes

Oestrous cyclicity (parental animals):
Oestrus cycle: Yes
Sperm parameters (parental animals):
Sperm parameters: testis weight, epididymis weight, enumeration of  homogenisation-resistant spermatids in testes, enumeration of cauda epididymal sperm reserve, sperm motility, sperm morphology
Litter observations:
Offspring: number and sex of pups, number of pubs per dam, offspring sex ratio, stillbirths, body weight gain, number of pubs with malformations,  functional development, physical development
Postmortem examinations (parental animals):
HISTOPATHOLOGY / ORGAN WEIGHTS
- Organ weights: Uterus, ovaries, testicles, epididymides,  prostate, seminal vesicles, brain, liver, kidneys, spleen, pituitary, thyroid, adrenal glands
- Histopathology: adrenal, coagulating gland, epididymide, ovary, pituitary, prostate, seminal vesicles, testicle, uterus with oviducts and cervix, vagina, grossly abnormal tissue
Postmortem examinations (offspring):
HISTOPATHOLOGY / ORGAN WEIGHTS
- Organ weights: Uterus, ovaries, testicles, epididymides,  prostate, seminal vesicles, brain, liver, kidneys, spleen, pituitary,  thyroid, adrenal glands
- Histopathology: adrenal,coagulating gland, epididymide, ovary,  pituitary, prostate, seminal vesicles, testicle, uterus with oviducts and cervix, vagina, grossly abnormal tissue
- Histopathology F1 not selected for mating, F2: grossly abnormal tissues  (if applicable)

Results and discussion

Results: P0 (first parental animals)

Details on results (P0)

Statistically significant reduction in food consumption was observed in  high dose males and females of the P- and F1-generation during the  pre-mating period and in high dose females also during gestation and  lactation period. Intermediate dosed dams showed a slight reduction in  food consumption during the pre-mating (P- and F1-generation) and the  gestation period (P-generation). The reduced food intake was related to  caloric substitution via sorbic acid. Reduced body weight and body weight gain was observed in high dose males  (P- and F1-generation). A significant dose-related reduction in body  weights of F1-pups was observed in the mid and high dose groups during  the lactation period. F2-pups of the high dose group showed also  significantly decreased body weights. However, this effect on body weight  was regarded as not adverse but related to the administration of an acid  at a high dose. In a 90-day dietary toxicity study no adverse effects on  body weight were recorded up to the highest tested doses of 6800 mg/kg  for males and 7200 mg/kg for females (IUCLID section 7.5.1). Treatment-related death occurred in F1 male and female parents  administered with sorbic acid at the high dose level. Animals of this  group showed also treatment-related clinical signs during the pre-mating  period (lateral position, reduced motility, pilo-erection and dyspnoea). Male fertility parameters, oestrus cycle length and normality as well as  reproductive performance were not influenced by the treatment (P- and  F1-generation).Cleavage of the balanopreputial gland and vaginal opening  were marginally but statistically significantly delayed in high dose  F1-pups selected for mating when compared to the control. The reduced anogenital distance found for F2-pups was related to the  slightly reduced body weight of the F2 pups during lactation and was thus  not regarded as an adverse effect. Gross pathology and histopathology revealed no changes related to  treatment for all animals examined. Statistically significant reduced absolute brain weights occurred in high dose F1-parent males and females and reduced absolute adrenal weights only in females. High dose F1-weanlings of both sexes showed decreased absolute brain and spleen weights and reduced fasting body weights. F2-pups showed decreased  absolute spleen weights and reduced body weights upon necropsy. Finally, it is noted that the extrapolation from sorbic acid to potassium sorbate is considered not to be restricted in any way, since the determinant of potential toxicity is the sorbate anion.

Effect levels (P0)

Dose descriptor:
NOAEL
Effect level:
3 000 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain
food consumption and compound intake

Results: F1 generation

Details on results (F1)

Statistically significant reduction in food consumption was observed in high dose males and females of the P- and F1-generation during the pre-mating period and in high dose females also during gestation and lactation period. Intermediate dosed dams showed a slight reduction in  food consumption during the pre-mating (P- and F1-generation) and the gestation period (P-generation). The reduced food intake was related to caloric substitution via sorbic acid. Reduced body weight and body weight gain was observed in high dose males (P- and F1-generation). A significant dose-related reduction in body weights of F1-pups was observed in the mid and high dose groups during  the lactation period. F2-pups of the high dose group showed also significantly decreased body weights. However, this effect on body weight was regarded as not adverse but related to the administration of an acid at a high dose. In a 90-day dietary toxicity study no adverse effects on  body weight were recorded up to the highest tested doses of 6800 mg/kg  for males and 7200 mg/kg for females (IUCLID section 7.5.1). Treatment-related death occurred in F1 male and female parents administered with sorbic acid at the high dose level. Animals of this group showed also treatment-related clinical signs during the pre-mating  period (lateral position, reduced motility, pilo-erection and dyspnoea). Male fertility parameters, oestrus cycle length and normality as well as reproductive performance were not influenced by the treatment (P- and  F1-generation). Cleavage of the balanopreputial gland and vaginal opening were marginally but statistically significantly delayed in high dose F1-pups selected for mating when compared to the control. The reduced anogenital distance found for F2-pups was related to the slightly reduced body weight of the F2 pups during lactation and was thus not regarded as an adverse effect. Gross pathology and histopathology revealed no changes related to treatment for all animals examined. Statistically significantly reduced absolute brain weights occurred in high dose F1-parent males and females and reduced absolute adrenal weights only in females. High dose F1-weanlings of both sexes showed decreased absolute brain and spleen weights and reduced fasting body weights. F2-pups showed decreased absolute spleen weights and reduced body weights upon necropsy. Finally, it is noted that the extrapolation from sorbic acid to potassium sorbate is considered not to be restricted in any way, since the determinant of potential toxicity is the sorbate anion.

Effect levels (F1)

open allclose all
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
1 000 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food consumption and compound intake
Dose descriptor:
LOAEL
Generation:
F1
Effect level:
3 000 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food consumption and compound intake

Results: F2 generation

Effect levels (F2)

open allclose all
Dose descriptor:
NOAEL
Generation:
F2
Effect level:
1 000 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain
organ weights and organ / body weight ratios
Dose descriptor:
LOAEL
Generation:
F2
Effect level:
3 000 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain
organ weights and organ / body weight ratios

Overall reproductive toxicity

Reproductive effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
- LO(A)EL
Parent animals: Not applicable.
F1-generation: 3000 mg/kg b.w./day

The extrapolation from sorbic acid to potassium sorbate or vice versa is considered not to be restricted in any way, since the determinant of potential toxicity is on the "sorbate" anion.
F2-generation: 3000 mg/kg b.w./day
- NO(A)EL
Parent animals: 3000 mg/kg b.w./day
F1-generation: 1000 mg/kg b.w./day
F2-generation: 1000 mg/kg b.w./day

The NOAEL for male and female animals of the P-generation was established at 3000 mg/kg bw/day. The statistically significant reduction of food intake in particular for F0 and F1 dams at 1000 and 3000 mg/kg bw/day, in the presence of caloric substitution by sorbic acid was considered to be the cause of reduced body weight development and slight developmental disturbances (morphological landmarks, learning and memory) of the F1/F2 offspring of the mid and high dose group during lactation. The reason for this effect remains unknown, however nutritional deficiencies in the pups masqued by caloric overcompensation in lactating females might be an explanation. The NOAEL concerning effects on development of the conceptus and the offspring (F1-generation) through sexual maturity was established at 1000 mg/kg bw/day. Unscheduled deaths and clinical signs in F1 weanlings selected for mating (observed for 5 juveniles at 3000 mg/kg bw/day and one juvenile at 1000 mg/kg bw/day) during early pre-mating period is not uncommon in oral (gavage) reproduction toxicity studies. When administering excessive dose levels to juveniles as in this case, intolerance to oral gavage treatment often is a more important aspect rather than toxic effects induced by the test compound itself - findings that would not be necessarily seen in a corresponding dietary (feeding) study. Hence, these deaths may also be considered as incidental and not treatment related, and therefore of no toxicological relevance.