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Toxicological information

Toxicity to reproduction

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Administrative data

Endpoint:
one-generation reproductive toxicity
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Justification for type of information:
Data is from peer-reviewed journal

Data source

Reference
Reference Type:
publication
Title:
Study of the teratogenic potential of FD & C yellow no. 5 when given by gavage to rats
Author:
T. F. X. COLLJNS, T. N. BLACK, L. H. BROWN and P. BULHACK
Year:
1990
Bibliographic source:
Food Chem Toxic, Vol. 28, No. 12, pp. 821-827

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
Principles of method if other than guideline:
To evalute the toxicity of Tartrazine in female Osborne-Mendel rats by gavage.
GLP compliance:
not specified
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: liquid
Details on test material:
- Name of test material (as cited in study report):Tartrazine
- Molecular formula :C16H12N4O9S2.3Na
- Molecular weight :534.36 g/mole
- Substance type:Organic
- Physical state:Liquid , dye
- Analytical purity:92.7%
- Impurities (identity and concentrations):No data available.
Specific details on test material used for the study:
- Name of test material (as cited in study report):Tartrazine
- Molecular formula :C16H12N4O9S2.3Na
- Molecular weight :534.36 g/mole
- Substance type:Organic
- Physical state:Liquid , dye
- Analytical purity:92.7%

Test animals

Species:
rat
Strain:
Osborne-Mendel
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: FDA rat breeding colony
- Age at study initiation: (P) female x wks; - 13-21 wk old, Male – No data available (F1) x wks
- Weight at study initiation: (P) Males: x-x g; - No data available. Females: x-x g;-210 -270 g (F1) Males: x-x g; No data available ,Females: x-x g- No data available
- Fasting period before study: No data available
- Housing: Housed in stainless-steel hanging cages.
- Diet (e.g. ad libitum): Purina Laboratory Chow, (Ralston Purina Company. St Louis, MO, USA) was available ad lib.
- Water (e.g. ad libitum): Distilled water was available ad lib.
- Acclimation period: No data available

ENVIRONMENTAL CONDITIONS
- Temperature (°C):2 0.5-25C
- Humidity (%):30-50%
- Air changes (per hr): No data available
- Photoperiod (hrs dark / hrs light): 12-hr light/dark cycle

Administration / exposure

Route of administration:
oral: gavage
Type of inhalation exposure (if applicable):
not specified
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:not specified

DIET PREPARATION
- Rate of preparation of diet (frequency):not specified
- Mixing appropriate amounts with (Type of food):not specified
- Storage temperature of food:not specified

VEHICLE
- Justification for use and choice of vehicle (if other than water):
- Concentration in vehicle: 0, 60, 100, 200, 400, 600 and 1000 mg/kg
- Amount of vehicle (if gavage): 1 ml/100g body weight
- Lot/batch no. (if required): not specified
- Purity not specified
Details on mating procedure:
- M/F ratio per cage:1:2
- Length of cohabitation: No data available
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy- The presence of sperm in the vaginal
smear was considered to be evidence of copulation and the sperm positive females were considered to be at day 0 of gestation.
- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility.-No data available
- Further matings after two unsuccessful attempts: [no / yes (explain)]- No data available
- After successful mating each pregnant female was caged (how):-No data available
- Any other deviations from standard protocol:No data available
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
19 days
Frequency of treatment:
Daily
Details on study schedule:
FD & C Yellow No. 5 (tartrazine) was given to Osborne-Mendel rats by gavage at dose levels of 0, 60,100, 200, 400, 600 and 1000mg/kg body weight/day on days 0-19 of gestation. Maternal or developmental toxicity was observed till the rats were killed on day 20. Fetal skeletal and visceral development was observed among fetuses from all groups.
Doses / concentrationsopen allclose all
Dose / conc.:
0 mg/kg bw/day
Dose / conc.:
60 mg/kg bw/day
Dose / conc.:
100 mg/kg bw/day
Dose / conc.:
200 mg/kg bw/day
Dose / conc.:
400 mg/kg bw/day
Dose / conc.:
600 mg/kg bw/day
Dose / conc.:
1 000 mg/kg bw/day
No. of animals per sex per dose:
Total no of animals- 259 female rats
0 mg/kg /day -38 female rats
60 mg/kg /day -37 female rats
100 mg/kg /day -37female rats
200 mg/kg /day -35 female rats
400 mg/kg /day -38 female rats
600 mg/kg /day -38 female rats
1000 mg/kg /day -36 female rats
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: 40-41 pregnant female randomly selected for dosing.
Positive control:
not specified

Examinations

Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS:Yes


DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule:Daily

BODY WEIGHT: Yes
- Time schedule for examinations:Daily

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No
- Time schedule for examinations:

OTHER:
Estrous cyclicity (parental animals):
Any irregularity in estrous cyclicity were observed
Sperm parameters (parental animals):
No data available.
Litter observations:
The foetuses were grouped by litter for identification. Each live foetus was promptly weighed, sexed and examined for gross external malformations, and the crown-rump length was measured. Any foetus that weighed less than 70% of the average weight of the concurrent male or female controls was considered to be a runt. External visceral and sternebral variations were also observed.
Postmortem examinations (parental animals):
On day 20 of gestation, the females were examined for gross abnormalities for the last time before being killed by CO2 asphyxiation. Caesarean sections were performed, corpora lutea were counted and the uteri were opened and examined in situ. The uterine positions of all implantation sites were noted and their condition (early or late resorptions, alive or dead foetuses) was determined.
Postmortem examinations (offspring):
Approximately half of the foetuses were examined for skeletal variations after being fixed in alcohol, cleared and stained with Alizarin Red S by a modification of Dawson's method. The remaining foetuses were fixed in Bouin's solution and sectioned according to the method of Wilson in order to detect internal visceral variations. Specific sternebral variation were observed in all fetuses like Incomplete ,Bipartite, Missing and Malaligned.Specific soft tissue variation like Hydroureter,Enlarged renal pclvis,Ectopic kidney ,Haemorrhages , Hydrocephalus ,Ectopic/incomplete descended testes ,Ectopic ovary ,Microphthalmia ,Anophthalmia ,Oedema and Defective heart valve were observed . Skeletal variation like 15 rib,l4th rib l3th rib ,l2th rib, Ribs, fused Rjbs, wavy ribs,Centra. red. on. Centra, misshapen ,Centra, bipartite ,Centra, missing Centra, fused wI dorsal arches ,Vertebrae. missing Caudal ossiBcation ,Dorsal arches, red Dorsal arches, fused Interparietal, red. oss. , Pariewl, red ,Frontal, red oss. ,Nasal red. oss. Supraoccipital, red. Oss,Supraoccipitaf. bipartite Hyoid. Red oss.Squuamosa, red. Zygomatic, red. ms. ,Pubis, red oss. Metacarpals, red. w Scapula. red oss. ,Scapula, curved ,
Clavicle, misshapen Maxillary, red. oss. ere observed .
Statistics:
Statistical method like ANOVA, Fishers exact test, t-test etc were used.
Reproductive indices:
Fertility index and delivery index were observed.
Offspring viability indices:
Viability index was observed.

Results and discussion

Results: P0 (first parental animals)

General toxicity (P0)

Clinical signs:
no effects observed
Description (incidence and severity):
No significant change were observed in behaviour and clinical sign.
Dermal irritation (if dermal study):
not specified
Mortality:
no mortality observed
Description (incidence):
No mortality was observed .Except one female in the group given 60 mg/kg body weight/day died at day 13 of gestation of gavage difficulties unrelated to dosage.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
No significant change were observed in body weight at all treated group 60, 100, 200, 400 and 600 mg/kg/day compare to control.
Except at dose group 1000 mg/kg/day, where body weight increased compare to control. Initial body weight at day 0 and maternal body-weight gain during gestation did not vary significantly between treated and control groups.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
No significant change were observed in food consumption at all treated group 60, 100, 200, 400 and 600 mg/kg/day compare to control.
Except at dose group 1000 mg/kg/day, where food consumption increased compare to control.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
no effects observed
Description (incidence and severity):
No significant change were observed in number of implants and number of resorption sites of all treated group at 60, 100, 200, 400, 600 and 1000 mg/kg /day as compare to control.
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
not specified
Histopathological findings: neoplastic:
not specified
Other effects:
not specified

Reproductive function / performance (P0)

Reproductive function: estrous cycle:
not specified
Reproductive function: sperm measures:
not specified
Reproductive performance:
no effects observed
Description (incidence and severity):
No significant change were observed in% pregnent rats and % implantation efficiency in all treated group at 60, 100, 200, 400, 600 and 1000 mg/kg /day as compare to control.

Effect levels (P0)

Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day
Based on:
test mat.
Sex:
female
Basis for effect level:
clinical signs
mortality
gross pathology
reproductive performance
Remarks on result:
other: No effect on reproductive performance

Target system / organ toxicity (P0)

Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified
Treatment related:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified

Results: F1 generation

General toxicity (F1)

Clinical signs:
not specified
Dermal irritation (if dermal study):
not specified
Mortality / viability:
no mortality observed
Description (incidence and severity):
No significant change were observed in the viability for male and female fetuses in all treated group compare to control.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
No significant changes were observed in the body weight of male and female fetuses in all treated group compare to control.
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Sexual maturation:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
no effects observed
Description (incidence and severity):
No significant change were observed in Specific sternebral variation all treated group60, 100, 200, 400, 600 and 1000 mg/kg /day compare to control.
No significant changes were observed in Specific Skeletal variation in all treated group60, 100, 200, 400, 600 and 1000 mg/kg /day compare to control.
Histopathological findings:
no effects observed
Description (incidence and severity):
No significant change were observed in Specific soft tissue variation like Hydroureter, Enlarged renal pclvis, Ectopic kidney, Haemorrhages , Hydrocephalus ,Ectopic/incomplete descended testes, Ectopic ovary, Microphthalmia, Anophthalmia, Oedema and Defective heart valve all treated group60, 100, 200, 400, 600 and 1000 mg/kg /day compare to control.
Other effects:
no effects observed
Description (incidence and severity):
No significant change were observed in the crown rump length of male and female foetuses in all treated group compare to control.

Developmental neurotoxicity (F1)

Behaviour (functional findings):
not specified

Developmental immunotoxicity (F1)

Developmental immunotoxicity:
not specified

Effect levels (F1)

Dose descriptor:
NOAEL
Generation:
F1
Effect level:
1 000 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
viability
mortality
body weight and weight gain
gross pathology
histopathology: non-neoplastic
other: No effect on crown rump length
Remarks on result:
other: No effect on viability, body weight, gross pathology and histopathology

Target system / organ toxicity (F1)

Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified
Treatment related:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified

Overall reproductive toxicity

Reproductive effects observed:
not specified
Treatment related:
not specified
Relation to other toxic effects:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified

Any other information on results incl. tables

To preclude the possibility of bias, all evaluations of the dams and foetuses during caesarean sections and during skeletal or visceral analysis were done without the evaluators knowing the dose-level group to which the animals belonged. Maternal and reproductive outcome at autopsy and foetal data at rats given F D &C yellow no.5 by gavage.

Applicant's summary and conclusion

Conclusions:
NOAEL was considered to be 1000 mg/kg bw/day for F0 and F1 generation when Osborne-Mendel female rats were treated with Tartrazine orally by gavage for 17 days.
Executive summary:

In a Teratogeninc potential study, Osborne-Mendel female rats were treated with Tartrazine in the concentration of 0, 60, 100, 200, 400, 600 and 1000 mg/kg /day orally by gavage in water. One female in 60 mg/kg body weight/day died on day 13 of gestation of gavage difficulties unrelated to dosage. No clinical sign of toxicity were observed in treated female rats as compared to control. Significant increase in food consumption at 1000 mg/kg body weight/day and no effect on body weight were observed in treated female rats as compared to control. No effect on % of pregnant female, no of corpora, implantation, no of viable foetuses, 5 resorptions, early death or late deaths per litter and sex ratio of treated rats as compared to control. Similarly, no effect on foetuses viability and body weight were observed in treated rats. In addition, three hydrocephalic pups in a single litter and isolated increase in the number of litters containing foetuses with at least two types of stemebral variations in foetuses at 200 mg/kg body weight/day, significant increase in haemorrhages at 600 mg/kg body weight/day and four foetuses from three litters at 1000 mg/kg body weight/day were observed but, The incidences of foetuses with visceral variations and of litters containing those foetuses were similar in all groups. Therefore, NOAEL was considered to be 1000 mg/kg bw/day for F0 and F1 generation when Osborne-Mendel female rats were treated with Tartrazine orally by gavage for 19 days.