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

Toxicity to reproduction

Administrative data

Endpoint:

three-generation reproductive 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: guidelines issued by ICH [12] and OECD [13] adapted for mice.

Principles of method if other than guideline:

Three-generation behavior toxicity study of tartrazine in mice.

GLP compliance:

not specified

Limit test:

no

Justification for study design:

No data available.

Test material

Test material information

Constituent 1

Reference substance name:

Tartrazine

IUPAC Name:

Tartrazine

Constituent 2

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 :C16H12N4O9S2.3Na
- 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 :C16H12N4O9S2.3Na
- 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 species / strain selection:

No data available.

Sex:

male/female

Details on test animals or test system and environmental conditions:

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:

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.


VEHICLE
- Justification for use and choice of vehicle (if other than water):No data available
- Concentration in vehicle: 0.05%, 0.15%, and 0.45%,
- Amount of vehicle (if gavage): Not applicable.
- Lot/batch no. (if required): No data available
- Purity: No data available

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.

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

approx. 200 days

Frequency of treatment:

Daily

Details on study schedule:

- F1 parental animals not mated until [...] weeks after selected from the F1 litters.:5 weeks of age
- Selection of parents from F1 generation when pups were [...] days of age.9 weeks of age
- Age at mating of the mated animals in the study: [...] weeks:4-9 weeks

Doses / concentrations

Remarks:

Doses / Concentrations:
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, 10 female
71.3 mg/kg bw/day -10 male, 10 female
214.9 mg/kg bw/day-10 male, 10 female
642.9 mg/kgbw/day -10 male, 10 female

Control animals:

yes, concurrent vehicle

Details on study design:

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

Positive control:

No data available.

Examinations

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: yes

DETAILED CLINICAL OBSERVATIONS: No data available.
- Time schedule:

BODY WEIGHT: Yes
- Time schedule for examinations:The 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):
- 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: Yes

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

OTHER: 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.

Oestrous cyclicity (parental animals):

No data available.

Sperm parameters (parental animals):

No data available.

Litter observations:

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%.
For F1 generation body weight was observed at 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, and 9 weeks of age after weaning. Different neurobehavioral test like Surface righting,
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. 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.
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.

Postmortem examinations (parental animals):

No data available.

Postmortem examinations (offspring):

No data available.

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

Reproductive indices:

- Fertility index was observed.

Offspring viability indices:

Viability index was observed.

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

effects observed, treatment-related

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):

No significant change was observed in the male 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 .

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

Histopathological findings: non-neoplastic:

not specified

Histopathological findings: neoplastic:

not specified

Other effects:

not specified

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

not specified

Reproductive function: sperm measures:

not specified

Reproductive performance:

effects observed, treatment-related

Description (incidence and severity):

Abortion was observed in four dams, one dam each in the low dosed(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.

Details on results (P0)

CLINICAL SIGNS (PARENTAL ANIMALS)- 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.

BODY WEIGHT - No significant change was observed in the male and female mice during the preconception or mating periods in all treated groupcompare 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 .

FOOD CONSUMPTION (PARENTAL ANIMALS)-- 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.

TEST SUBSTANCE INTAKE (PARENTAL ANIMALS)-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.

REPRODUCTIVE FUNCTION: ESTROUS CYCLE (PARENTAL ANIMALS)-No data available

REPRODUCTIVE FUNCTION: SPERM MEASURES (PARENTAL ANIMALS)- No data available

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)-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.


ORGAN WEIGHTS (PARENTAL ANIMALS)- No data available

GROSS PATHOLOGY (PARENTAL ANIMALS)- No data available

HISTOPATHOLOGY (PARENTAL ANIMALS)-No data available

OTHER FINDINGS (PARENTAL ANIMALS)- No data available

Effect levels (P0)

Dose descriptor:

NOAEL

Effect level:

642.8 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

water consumption and compound intake

reproductive performance

other: No significant effect were observed on the clinical sign,body weight ,food consumption,chemical intake and reproductive performace .

Remarks on result:

other: No treatment related effects was observed

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:

effects observed, treatment-related

Dermal irritation (if dermal study):

not specified

Mortality / viability:

mortality observed, non-treatment-related

Description (incidence and severity):

One dam killed its litter in the high-dosed group during the first week of the lactation. All offspring of one litter died in the control group during
the first weeks of the lactation period since their dam had not nursed.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

In the , 0.05% dosed group, the average body weight of male and female offspring was increased significantly throughout the lactation period .In the 0.15%dosed group, the average body weight of male offspring was increased significantly at PND 7.In the 0.45% dosed group, the average body weight of male offspring was increased significantly at PND 21. 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.

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 F1 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):

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Sexual maturation:

effects observed, treatment-related

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

not specified

Histopathological findings:

not specified

Other effects:

not specified

Developmental neurotoxicity (F1)

Behaviour (functional findings):

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

Developmental immunotoxicity (F1)

Developmental immunotoxicity:

not specified

Details on results (F1)

VIABILITY (OFFSPRING)-
F1 generation
NO significant change were observed in the viability index in F1 generation male and female offspring compare to control during lactation period.

F2generation - 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.

CLINICAL SIGN
F1 generation
Clinical signs- No significant change were observed in thelitter size and sex ratio at birth in F1 generation male and female offspring compare to control during lactation period.
F2generation - No significant adverse effect was observed in litter size, litter weight, or sex ratio at birth.

Sexual maturation- (OFFSPRING)-
F1 generation
Reproductive performance- Four dams had not became pregnant, two dams in the control and one dam each in the low-dosed (0.05%) and middle-dosed groups(0.15). Abortion was observed in one dam in the high-dosed group(0.45%).

BODY WEIGHT (OFFSPRING)
F1 generation
Body weight -No significant change was observed in the male and female mice during the preconception or mating periods in all treated group compare 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.

food consumption- No significant change was observed in the food consumption of F1 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.

F2 generation
Body weight -
In the low-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 middle-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
.
food consumption- No significant change was observed in the food consumption of F2 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.


ORGAN WEIGHTS (OFFSPRING) No data available

GROSS PATHOLOGY (OFFSPRING)-No data available

HISTOPATHOLOGY (OFFSPRING)-No data available

OTHER FINDINGS (OFFSPRING)-
F1 generation
Neurological behaviour-
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.

F2generation -
Neurological behaviour-
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.
Test substance intake
F1 generation
Test substance intake- No significant change was observed in the chemical intake of F1 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.
F12generation
Test substance intake- No significant change was observed in the chemical intake of F2 male and female mice during the preconception ,mating periods, gestation and lactation period in all treated group 0, 0.05%, 0.15%, and 0.45% compare to control.

Effect levels (F1)

1

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

642.8 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

viability

mortality

body weight and weight gain

food consumption and compound intake

developmental neurotoxicity

other: see 'Remark'

Remarks on result:

other: No reproductive toxicity was observed

2

Dose descriptor:

LOAEL

Generation:

F1

Effect level:

214.9 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

not specified

Basis for effect level:

body weight and weight gain

other: neurobehavioural changes

Remarks on result:

other: neurobehavioural changes was observed

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

Results: F2 generation

General toxicity (F2)

Clinical signs:

not specified

Dermal irritation (if dermal study):

not specified

Mortality / viability:

not specified

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

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.

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 F2 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 F2 male and female mice during the preconception ,mating periods, gestation and lactation period in all treated group 0, 0.05%, 0.15%, and 0.45% compare to control.

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:

not specified

Histopathological findings:

not specified

Other effects:

not specified

Developmental neurotoxicity (F2)

Behaviour (functional findings):

effects observed, treatment-related

Description (incidence and severity):

The behavioural developmental parameters, the development of swimming direction at PND 7 was accelerated significantly in the 0.45%dosed group in male offspring. Time taken of olfactory orientation at PND 14 was accelerated significantly in the 0.15%-dosed and 0.45% dosed groups and those effects were significantly dose-related in a trend test(P < 0.01). Surface righting at PND 7 was accelerated significantly in the 0.15% dosed group in female offspring . In movement activity of exploratory behaviour at 3 weeks of age, total distance (cm), average distance (cm) and average speed (cm/s) showed a significant tendency to be affected in the treatment groups in male offspring in a trend test (P < 0.05, 0.05 and 0.01, respectively.

Developmental immunotoxicity (F2)

Developmental immunotoxicity:

not specified

Effect levels (F2)

1

Dose descriptor:

NOAEL

Generation:

F2

Effect level:

642 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

not specified

Basis for effect level:

mortality

body weight and weight gain

food consumption and compound intake

Remarks on result:

other: No developmental effects was observed

2

Dose descriptor:

LOAEL

Generation:

F2

Effect level:

214.9 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

not specified

Basis for effect level:

body weight and weight gain

other: neurobehavioural changes

Remarks on result:

other: effects on neurobehavioural changes observed

Target system / organ toxicity (F2)

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

Applicant's summary and conclusion

Conclusions:

In three generation reproductive toxicity study,NOAEL was considered to be 642.8 mg/kg/day (0.45%) for Tartrazine in both P and F1 generation (Crlj: CD1 male and female mice by oral diet.

Executive summary:

A three generation reproductive toxicity study was conducted forTartrazine(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.

 

In parents,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 the male 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 be642.8 mg/kg/day (0.45%) for Tartrazine(1934-21-0) in both P and F1 generation (Crlj: CD1 )male and female mice for reprotoxicity effect.