Aluminium, 4,5-dihydro-5-oxo-1-(4-sulfophenyl)-4-[(4-sulfophenyl)azo]-1H-pyrazole-3-carboxylic acid complex

Developmental toxicity / teratogenicity

Administrative data

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Justification for type of information:

Data is from peer-reviewed journal

Data source

Reference

Reference Type:

publication

Title:

Effects of tartrazine on exploratory behavior in a three-generation toxicity study in mice.

Author:

Toyohito Tanaka∗, Osamu Takahashi, Shinshi Oishi, Akio Ogata

Year:

2008

Bibliographic source:

Reproductive Toxicology 26 , 156–163,2008

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: OECD Guideline 426 (Developmental Neurotoxicity Study)

Principles of method if other than guideline:

Three-generation reproductive and neurobehavioral toxicity study of Tartrazine in male and female mice

GLP compliance:

not specified

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Trisodium 5-hydroxy-1-(4-sulphophenyl)-4-(4-sulphophenylazo)pyrazole-3-carboxylate

EC Number:

217-699-5

EC Name:

Trisodium 5-hydroxy-1-(4-sulphophenyl)-4-(4-sulphophenylazo)pyrazole-3-carboxylate

Cas Number:

1934-21-0

Molecular formula:

C16-H12-N4-O9-S2.3Na

IUPAC Name:

trisodium 5-hydroxy-1-(4-sulphophenyl)-4-(4-sulphophenylazo)pyrazole-3-carboxylate

Test material form:

solid: particulate/powder

Details on test material:

- Name of test material (as cited in study report):Tartrazine
- Molecular formula:C16H12N4O9-S2.3Na
- Molecular weight :534.36 g/mole
- Substance type:Organic
- Physical state:Liquid , dye
- Analytical purity:85.0%
- 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:C16H12N4O9-S2.3Na
- Molecular weight :534.36 g/mole
- Substance type:Organic
- Physical state:Liquid , dye
- Analytical purity:85.0%
- Impurities (identity and concentrations):No data available.

Test animals

Species:

mouse

Strain:

Crj: CD(SD)

Details on test animals or test system and environmental conditions:

Details on test animal
TEST ANIMALS
- Source: Charles River Japan Inc., Kanagawa, Japan
- Age at study initiation: 5 weeks of age of the F0 generation to 9 weeks of age of the F2 generation.
- Weight at study initiation: (P) Males: x-x g; Females: x-x g; (F1) Males: x-x g; Females: x-x g
- Fasting period before study: No data available.
- Housing: They were housed individually in polycarbonate
Solid-floored cages with wood flakes.
- Diet (e.g. ad libitum): Basal diets (Nihon Clea, CE-2)
ad libitum.
- Water (e.g. ad libitum): They were given water ad libitum.
- Acclimation period: 1 week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 25±1 ◦C
- Humidity (%):50±5%.
- Air changes (per hr): No data available
- Photoperiod (hrs dark / hrs light): 12 h light/12 h dark cycle

Administration / exposure

Route of administration:

oral: feed

Vehicle:

other: Basal diets (Nihon Clea, CE-2)

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:

DIET PREPARATION
- Rate of preparation of diet (frequency): The experimental diets were prepared bimonthly (three times) in the laboratory.
- Mixing appropriate amounts with (Type of food): After mixing tartrazine with the powdered diet (basal Diets, Nihon Clea, CE-2), pellets were formed and fed to mice. Tartrazine was stable in the pellets during the experimental period when previously measured by
HPLC. The homogeneity of the test compound was ensured by the preparation procedures of the experimental diets in the laboratory when previously measured by HPLC. The concentration and homogeneity of the test compound in the diet was not tested during the experimental period.

- Storage temperature of food: No data available.

Analytical verification of doses or concentrations:

not specified

Details on mating procedure:

- M/F ratio per cage:1:1
- Length of cohabitation: 5 days
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy-No data available.
- 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 protocolNo data available.

Duration of treatment / exposure:

199 days (approx 200)

Frequency of treatment:

Daily

Duration of test:

199 days (approx 200)

Doses / concentrations

Remarks:

0, 0.05%, 0.15%, and 0.45% (0,71.3, 214.9 and 642.8 mg/kg/day)

No. of animals per sex per dose:

Total no. of animals-80
0 mg/kg bw/day -10 male and 10 female
71.3 mg/kg bw/day -10 male and 10 female
214.9 mg/kg bw/day -10 male and 10 female
642.9 mg/kg/day -10 male and 10 female

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale:
Micewere assigned to the groups by the stratified randomization method.

Examinations

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule:
- Cage side observations checked in table [No.?] were included.

DETAILED CLINICAL OBSERVATIONS: Yes / No / No data
- Time schedule:

BODY WEIGHT: Yes
- Time schedule for examinations:he animals were weighed individually on experimental days 0, 2, 4, 7, 14, 21, 28, and 30 during the preconception period.

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

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

POST-MORTEM EXAMINATIONS: Yes / No / No data
- Sacrifice on gestation day #
- Organs examined:

OTHER:

Ovaries and uterine content:

No data available.

Fetal examinations:

- External examinations: Yes:
- Soft tissue examinations: No data
- Skeletal examinations: No data
- Head examinations: No data
Litter observation- For F1 generation offspring were weighed individually on PNDs 0, 4, 7, 14, and 21 during the lactation period. The survival indices were calculated as (live offspring at each period)/ (live and dead offspring at birth) ×100%. Negative geotaxis, Cliff avoidance, Swimming behavior and olfactory orientation were observed on PND 4-14 .Food intake and chemical intake was observed during Preconception, mating, gestation and
Lactation.
For F2 generation offspring were weighed individually on PNDs 0, 4, 7, 14, and 21 during the lactation period. The survival indices were calculated as (live offspring at each period)/ (live and dead offspring at birth) ×100%. The offspring were weaned when they were 4 weeks of age, and one male and one female were selected at random from each litter to continue treatment. The animals were weighed individually at 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, and 9 weeks of age after weaning.
Animals at 7weeks of age in the F1 and F2 generation each performed 1 trial per day for 3 days in a Biel-type multiple-T water maze adapted for mice. Exploratory behavior of mice was measured in an animal movement analyzing system ANIMATE AT- at 8 weeks of in the F1 and F2.

Statistics:

Food intake, litter size, litter weight, and body weight were assessed with Bonferroni’s multiple comparison tests after the analysis of variance (ANOVA) or the Kruskal–Wallis test. Sex ratio, survival and behavioural developmental data were assessed with the X2-test or with Fisher’s exact test of frequency analysis. Movement activity data were assessed with the Steel–Dwass test of non-parametric methods. Multiple-T water maze performance data were assessed with the Sign–Wilcoxon test for trials and assessed with the Steel–Dwass test within each treatment group. Dose-response effects were assessed with the Jonckheere test for ordered alternatives or the cumulative -test (multi) for frequency data.

Indices:

Fertility index and viability index of offspring were opbserved.

Historical control data:

No data available.

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:

no effects observed

Description (incidence and severity):

In movement activity of exploratory behavior at 8weeks of age, no variables of measurement showed a significant adverse effect of tartrazine in male and female mice of treated group compare to contol.

Dermal irritation (if dermal study):

not specified

Mortality:

not specified

Body weight and weight changes:

no effects observed

Description (incidence and severity):

The average body weight of dams showed no significant adverse effect during the gestation or lactation period in treated group compare to control

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

No significant change was observed in the food consumption of male and female mice during the preconception ,mating periods, gestation and lactation period in all treated group compare 0, 0.05%, 0.15%, and 0.45% to control

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

no effects observed

Description (incidence and severity):

No significant change was observed in the chemical intake of male and female mice during the preconception ,mating periods, gestation and lactation period in all treated group compare 0, 0.05%, 0.15%, and 0.45% to control.

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:

not specified

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

not specified

Histopathological findings: neoplastic:

not specified

Other effects:

not specified

Maternal developmental toxicity

Number of abortions:

effects observed, treatment-related

Description (incidence and severity):

Abortion was observed in four dams, one dam each in the (0.05%) dosedand (0.15%)dosed groups and two dams in the (0.45%) dosed groups

Pre- and post-implantation loss:

not specified

Total litter losses by resorption:

not specified

Early or late resorptions:

not specified

Dead fetuses:

not specified

Changes in pregnancy duration:

not specified

Description (incidence and severity):

Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): not specified

Changes in number of pregnant:

not specified

Other effects:

not specified

Details on maternal toxic effects:

Maternal toxic effects:no effects

Details on maternal toxic effects:
Body weight -No significant change was observed in the male and female mice during the preconception or mating periods in F0 generation compare to control.

Food consumption:No significant effect on food consumption were observed in treated mice in F0 generation as compared to control.

Test substance intake- No significant change was observed in the chemical intake of male and female mice during the preconception ,mating periods, gestation and lactation period in F0 generation as compared to control.

Reproductive performance- No significant adverse effect was observed on survival, litter size, litter weight, or sex ratio at birth were observed in F0 generation as compared to control.

Effect levels (maternal animals)

Dose descriptor:

NOAEL

Effect level:

642.8 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

other: developmental toxicity

Remarks on result:

other: No effects on developmental parameters

Maternal abnormalities

Abnormalities:

not specified

Results (fetuses)

Fetal body weight changes:

no effects observed

Description (incidence and severity):

No significant change was observed in the male and female mice during the preconception or mating periods in all treated groupcompare to control in F1 generation.The average body weight of dams showed no significant adverse effect during the gestation or lactation period in all treated group 0, 0.05%, 0.15%, and 0.45% compare to control.There was no sex-related difference in average body weight during the lactation period.

Reduction in number of live offspring:

no effects observed

Description (incidence and severity):

No significant adverse effect was observed in Viability index in all treated group 0, 0.05%, 0.15%, and 0.45%compare to control during lactation period.

Changes in sex ratio:

no effects observed

Description (incidence and severity):

No significant adverse effect was observed in sex ratio at birth

Changes in litter size and weights:

no effects observed

Description (incidence and severity):

No significant adverse effect was observed in litter size, litter weight at birth

Changes in postnatal survival:

not specified

External malformations:

not specified

Skeletal malformations:

not specified

Visceral malformations:

not specified

Other effects:

effects observed, treatment-related

Description (incidence and severity):

Significant change were observed in the neurobehaviur test ,Swimming direction at PND7 in F1 generation male at dose level 0.15%and surface righting at PND7 in female offspring at dose level 0.15%compare to control.Significant change were observed in the neurobehaviur test ,Swimming direction at PND7 and time taken of olfactory orientation at PND 14 in F2 generation male at dose level 0.15% and 0.45% respectively and surface righting at PND7 in female offspring at dose level 0.15 %compare to control.

Effect levels (fetuses)

1

Dose descriptor:

NOAEL

Effect level:

643.8 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No significant effect were observed onthe litter size or sex ratio at birth,litter weight and viability index of offspring

Remarks on result:

other: No developmental toxic effects was observed

2

Dose descriptor:

LOAEL

Effect level:

214.9 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

not specified

Basis for effect level:

other: neurobehavioural changes

Remarks on result:

other: neurobehavioural changes was observed

Fetal abnormalities

Abnormalities:

not specified

Overall developmental toxicity

Developmental effects observed:

not specified

Treatment related:

not specified

Relation to maternal toxicity:

not specified

Dose response relationship:

not specified

Relevant for humans:

not specified

Applicant's summary and conclusion

Conclusions:

NOAEL was found to be 642.8 mg/kg/day (0.45%) for Tartrazine in both F1and F 2generation (Crlj: CD1 male and female mice when their dams were exposed to test chemical by oral diet.

Executive summary:

A three generation reproductive toxicity study was conducted for Tartrazine(1934-21-0) in (Crlj: CD1 male and female mice.The design of the study was based on the guidelines issued by ICH[12]and OECD[13]adapted for mice The test materialwas given to male and female mice in the diet at levels of 0 (control), 0.05%, 0.15%, and 0.45%(0, 71.3, 214.9 and 642.8 mg/kg/day)from 5 weeks of ageof the F0 generation to 9weeks of age of the F2 generation. 10 male and 10 female in each dose group.Clinical sign ,body weight, food consumption,chemical intake ,reproductive and neurobehavioral parameters were measured. Thelitter size or sex ratio at birth,litter weight and viability index of offspring were also observed..No. of females examined, No. of pregnant females, No. of litters No. of offspring, Average litter size, Average litter weight, Total sex ratio (male/female) and Average sex ratio (male %) were also observed.Exploratory behavior of mice was measured in an animal movement analyzing system ANIMATE AT- at 8 weeks of in the F0.

 

InParents,movement activity of exploratory behavior at 8 weeks of age, no variables of measurement showed a significant adverse effect of tartrazine in male and female mice of treated group compare to contol.No significant change was observed in themale and female mice during the preconception or mating periods in all treated group compare to control.Even the average body weight of dams showed no significant adverse effect during the gestation or lactation period in treated group compare to control.No significant change was observed in the food consumption ofmale and female mice during the preconception, mating periods, gestation and lactation period in all treated group compare 0,0.05%, 0.15%, and 0.45%to control.No significant change was observed in the chemical intake ofmale and female mice during the preconception, mating periods, gestation and lactation period in all treated group compare 0,0.05%, 0.15%, and 0.45%to control. Abortion was observed in four dams, one dam each in the lowdosed(0.05%) and middle-dosed groups(0.15), and two dams in the high-dosed groups (0.45%). One dam in the high-dosed group(0.45%)killed its all offspring during the first week of lactation. One dam in the control(0%) group had not nursed its offspring during the first week of lactation.

 

In the F1 generation,In the,0.05%dosed group, the average body weight of male and female offspring was increased significantly throughout the lactation period .In the0.15%dosed group, the average body weight of male offspring was increased significantly at PND 7.In the0.45%dosed group, the average body weight of male offspring was increased significantly at PND 21.No significant change was observed in themale and female mice during the preconception or mating periods in all treated group compare to control in F1 generation.

The averagebody weight of dams showed no significant adverse effect during the gestation or lactation period in all treated group0,0.05%, 0.15%, and 0.45% compare to control.No significant change was observed in the food consumption of F1male and female mice during the preconception ,mating periods, gestation and lactation period in all treated group compare 0,0.05%, 0.15%, and 0.45%to control.The development of swimming direction at postnatal day (PND) 7 was accelerated significantly in male offspring in a dose-related manner. Surface righting at PND 7 was affected significantly in female offspring in dose-related manner. Several variables in exploratory behavior showed significant tendencies to be affected in the treatment groups in male offspring at 3 weeksof age.

 

In the F2 generation, In the 0.05% dosed group, the average body weight of female offspring was affected significantly at birth and was increased significantly at PNDs 14 and 21, and that of male offspring was increased significantly at PNDs 7, 14 and 21. In 0.15% dosed group, the average body weight of male offspring was increased significantly throughout the lactation period, and that of female offspring was increased significantly at PNDs 7, 14 and 21. There was no sex-related difference in average body weight during the lactation period.No significant change was observed in the food consumption of F2male and female mice during the preconception, mating periods, gestation and lactation period in all treated group compare 0,0.05%, 0.15%, and 0.45%to control.No significant change was observed in the chemical intake of F2male and female mice during the preconception, mating periods, gestation and lactation period in all treated group 0,0.05%, 0.15%, and 0.45%compareto control.The development of swimming direction at PND 7was accelerated significantly in the 0.45%dosed group in male offspring. Time taken of olfactory orientation at PND 14 was accelerated significantly in male offspring in a dose-related manner. Several variables in exploratory behavior showed significant tendencies to be affected in the treatment groups in male offspring at 3 weeks of age, and in males at 8 weeks of age. The dose levels of tartrazine in the present study produced a few adverse effects on neurobehavioral parameters throughout generations in mice .But no adverse effect were observed on reproductive parameter in all generations of (Crlj: CD1 male and female mice in all treated group 0,0.05%, 0.15%, and 0.45% compareto control. Therefore NOAEL was considered to be 642.8 mg/kg/day (0.45%) for Tartrazine(1934-21-0) in both F1and F 2 generation (Crlj: CD1 male and female mice for developmental effect.