Trisodium 3-hydroxy-4-(4'-sulphonatonaphthylazo)naphthalene-2,7-disulphonate

Developmental toxicity / teratogenicity

Administrative data

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: data from peer - reviewed journals

Data source

Reference

Reference Type:

publication

Title:

Effects of amaranth on F1 generation mice

Author:

Toyohito Tanaka

Year:

1992

Bibliographic source:

Toxicology Letters, 60 (1992) 315-324

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

other: as mentioned below

Principles of method if other than guideline:

To determine the reproductive toxicity of the test chemical in mice

GLP compliance:

not specified

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Trisodium 3-hydroxy-4-(4'-sulphonatonaphthylazo)naphthalene-2,7-disulphonate

EC Number:

213-022-2

EC Name:

Trisodium 3-hydroxy-4-(4'-sulphonatonaphthylazo)naphthalene-2,7-disulphonate

Cas Number:

915-67-3

Molecular formula:

C20H14N2O10S3.3Na

IUPAC Name:

trisodium 3-hydroxy-4-[(4-sulfonato-1-naphthyl)diazenyl]naphthalene-2,7-disulfonate

Constituent 2

Reference substance name:

trisodium (4E)-3-oxo-4-[(4- sulfonato-1- naphthyl)hydrazono]naphthalene- 2,7-disulfonate

IUPAC Name:

trisodium (4E)-3-oxo-4-[(4- sulfonato-1- naphthyl)hydrazono]naphthalene- 2,7-disulfonate

Constituent 3

Reference substance name:

Amaranth dye

IUPAC Name:

Amaranth dye

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

Name of test material (as cited in study report): Amaranth dye
Molecular formula (if other than submission substance): C20H11N2Na3O10S3
Molecular weight (if other than submission substance): 604.47
Smiles notation (if other than submission substance): c1ccc2c(c1)c(ccc2S(=O)(=O)[O])N=Nc3c4ccc(cc4cc(c3O)S(=O)(=O)[O-])S(=O)(=O)[O-].[Na+].[Na+].[Na+]
InChl (if other than submission substance): 1S/C20H14N2O10S3.3Na/c23-20-18(35(30,31)32)10-11-9-12(33(24,25)26)5-6-13(11)19(20)22-21-16-7-8-17(34(27,28)29)15-4-2-1-3-14(15)16;;;/h1-10,23H,(H,24,25,26)(H,27,28,29)(H,30,31,32);;;/q;3*+1/p-3
Substance type: Organic
Physical state: Solid

Test animals

Species:

mouse

Strain:

other: Crj:CD 1

Details on test animals or test system and environmental conditions:

Source: Charles River Japan inc (Tokyo,Japan)
Age at study initiation: 9 weeks at the start of study)
Weight at study initiation: No data
Fasting period before study: No data
Housing: individually housed in polycarbonate solid- floored cages with wood flakes
Diet (e.g. ad libitum): experimental or control diets, ad libitum
Water (e.g. ad libitum):Tap water, ad libitum
Acclimation period: No data
ENVIRONMENTAL CONDITIONS
Temperature (°C): No data
Humidity (%): No data
Air changes (per hr):No data
Photoperiod (hrs dark / hrs light):
No data
IN-LIFE DATES: From: To: No data

Administration / exposure

Route of administration:

oral: feed

Vehicle:

other: basal diet

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
Amaranth was administered in the diet to 60 mice (I0/sex/group) at dietary levels of 0.03,0.09 and 0.27%.

Analytical verification of doses or concentrations:

not specified

Details on analytical verification of doses or concentrations:

no data

Details on mating procedure:

M/F ratio per cage: Each female were mated with one male rat
Length of cohabitation: 5 days

Duration of treatment / exposure:

15 weeks

Frequency of treatment:

daily

Duration of test:

15 weeks

No. of animals per sex per dose:

60 mice (10/sex/dietary group)

Control animals:

yes, plain diet

Details on study design:

Dose selection rationale:
The dose levels were introduced on the basis of ADI of Amaranth dye (0.5 mg/kgb.w)
Other: The 20 mice (10/sex) in the control group were given the basal diet (Nihon Clea, CE-2) for a corresponding period of time.

Examinations

Maternal examinations:

At 8 weeks of age, the motor activity of the F0 generation mice was measured by an animal movement analyzing system. The behavioral parameters were recorded for 5 min. i.e. number of movements, movement time (s), number of horizontal movements, total distance (cm), number of vertical movements, vertical time (s), turning, average distance (cm), average speed (cm/s), and maximum continuation time (s)

Ovaries and uterine content:

no data

Fetal examinations:

The physical and functional developmental parameters were measured for individual pups, and were analyzed on a whole-litter basis as follows:
(I) Surface righting (PND 4 and 7)
(2) Negative geotuxis (PND 4 and 7)
(3) Cliff avoidance (PND 7)
(4) Swimming (PND 4, 14)
5) Olfactory orientation (PND 14)

Statistics:

Student’s t-test was performed on litter size, pup weight, and litter weight. The behavioral parameters were assessed with the Mann-Whitney U-test.

Indices:

no data

Historical control data:

no data

Results and discussion

Results: maternal animals

Maternal developmental toxicity

Details on maternal toxic effects:

Maternal toxic effects:no effects

Details on maternal toxic effects:
There was no consistent significant compound- or dose-related effect in motor activity of mice administered amaranth

Effect levels (maternal animals)

Remarks on result:

not determinable

Results (fetuses)

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:yes

Details on embryotoxic / teratogenic effects:
There were no consistent significant differences in the parameters of the litters. In the high-dose group, there were a few litters of small size. The body weight of the pups in the treatment groups during the lactation period was not significantly different from that of controls. The survival index at PND 21for male pups was control 88.9%, low-dose 76.0%, medium-dose 82.5%, and high dose 75.0%, and for female pups was control 88.9%, low-dose 74.8%, medium-dose 88.9%, and high-dose 68.3%. The developmental parameters were repressed in the amaranth 0.03% group (about 50 mg/kg/d)

Effect levels (fetuses)

Dose descriptor:

NOEL

Effect level:

50 mg/kg bw/day (nominal)

Based on:

test mat.

Remarks on result:

other: effects on some developmental and behavioral parameters observed in higher dose levels

Fetal abnormalities

Abnormalities:

not specified

Overall developmental toxicity

Developmental effects observed:

not specified

Any other information on results incl. tables

Table 1: Summary of data of litters in F1 generation study of Amaranth Administered in mice

Dose Levels (%)	0	0.03	0.09	0.27
No of litter	9	10	9	10
No of pups	109	124	109	97
Litter Size	12.1 ±0.93	12.4±2.80	12.1± 2.67	9.7±4.55
Pup Weight (g)	1.57±0.152	1.54±0.135	1.54±0.150	1.54 ±0.172
Litter Weight (g)	18.96±1.23	19.12±3.79	18.68±3.50	14.95±6.33

Eacch value represents mean± S.D

Applicant's summary and conclusion

Conclusions:

The color additive, amaranth, was given in the diet to provide dietary levels of 0 (control), 0.03,0.09 and
0.27%, from 5 weeks of age in F0, generation mice to 9 weeks of age in F1, generation mice, and some
reproductive, developmental and behavioral parameters were measured.

There was no effect on the parameters of litters, litter size, pup weight and litter weight. The body weight of pups during the lactation period in the treatment groups increased less significantly, and the survival index at postnatal day (PND) 21 of the amaranth 0.27% group was reduced. The developmental parameters were repressed in the amaranth 0.03% group (about 50 mg/kg/d)
The dose levels of amaranth influenced some reproductive, developmental and behavioral parameters in mice.
The No Observed Effect Level in maternal and F1 generation mice after administration of Amaranth dye was 0.03% (about 50 mg/kg/day).

Executive summary:

The color additive, amaranth, was given in the diet to provide dietary levels of 0 (control), 0.03,0.09 and

0.27%, from 5 weeks of age in F0, generation mice to 9 weeks of age in F1, generation mice, and some

reproductive, developmental and behavioral parameters were measured.

 

Male and female mice (Crj:CD-1, 4 weeks old) were obtained from Charles River Japan Inc. (Tokyo, Japan). They were individually housed in polycarbonate solid floored cages with wood flakes. They were given control or experimental diets and water ad libitum. Amaranth was administered in the diet to 60 mice (IO/sex/group) at dietary levels of 0.03, 0.09 and 0.27%. The 20 mice (lo/sex) in the control group were given the basal diet (Nihon Clea, CE-2) for a corresponding period of time The animals were 5 weeks old at the start of the study. At 8 weeks of age, the motor activity of the mice was measured by an animal movement analyzing system (ANIMATE AT-420, Toyo Sangyo Co., Ltd, Toyama, Japan).At 9 weeks of age, each female was paired with one male from the same treatment group, for a period of 5 days. After 5

days, the males were removed from the females, and the females were allowed to produce and rear their pups.

 

F1 generation

On postnatal day 0 (PND 0), litter size, pup weight, and litter weight were measured. The pups were weighed on PND 4, 7, 14 and 21 during the lactation period. The physical and functional developmental parameters were measured for individual pups, and were analyzed on a whole-litter basis as follows:

(I)Surface righting (PND 4 and 7)

(2)Negative geotuxis (PND 4 and 7)

(3) Cliff avoidance (PND 7)

(4)Swimming (PND 4, 14)

5)Olfactory orientation (PND 14)

Student’s t-test was performed on litter size, pup weight, and litter weight. The behavioral parameters were assessed with the Mann-Whitney U-test.

 There was no consistent significant compound- or dose-related effect in motor activity of mice administered amaranth.There was no effect on the parameters of litters, litter size, pup weight and litter weight. The body weight of pups during the lactation period in the treatment groups increased less significantly, and the survival index at postnatal day (PND) 21 of the amaranth 0.27% group was reduced.The developmental parameters were repressed in the amaranth 0.03% group (about 50 mg/kg/d)

The dose levels of amaranth influenced some reproductive, developmental and behavioral parameters in mice.

The No Observed Effect Level in maternal and F1 generation mice after administration of Amaranth dye was0.03% (about 50 mg/kg/day).