Disodium 6-hydroxy-5-[(2-methoxy-4-sulphonato-m-tolyl)azo]naphthalene-2-sulphonate

Toxicity to reproduction

Administrative data

Endpoint:

two-generation reproductive toxicity

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

secondary literature

Justification for type of information:

Data from secondary source

Data source

Reference

Reference Type:

other: Secondary source

Title:

Two generation reproductive toxicity study of test chemical

Author:

HPVIS

Year:

2018

Bibliographic source:

HPVIS,2018

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 416 (Two-Generation Reproduction Toxicity Study)

Principles of method if other than guideline:

Two generation reproductive toxicity study of test chemical

GLP compliance:

not specified

Limit test:

no

Justification for study design:

No data available

Test material

Test material information

Constituent 1

Reference substance name:

Disodium 6-hydroxy-5-[(2-methoxy-4-sulphonato-m-tolyl)azo]naphthalene-2-sulphonate

EC Number:

247-368-0

EC Name:

Disodium 6-hydroxy-5-[(2-methoxy-4-sulphonato-m-tolyl)azo]naphthalene-2-sulphonate

Cas Number:

25956-17-6

Molecular formula:

C18H16N2O8S2.2Na

IUPAC Name:

disodium 6-hydroxy-5-[(2-methoxy-3-methyl-4-sulfonatophenyl)diazenyl]naphthalene-2-sulfonate

Test material form:

solid: particulate/powder

Details on test material:

- Name of test material: disodium 6-hydroxy-5-[(2-methoxy-4-sulphonato-m-tolyl)azo]naphthalene-2-sulphonate (Allura red AC)
- IUPAC name: disodium 6-hydroxy-5-[(2-methoxy-3-methyl-4-sulfonatophenyl)diazenyl]naphthalene-2-sulfonate
- Moleclar formula: C18H16N2O8S2.2Na
- Molecular formula: 496.4266 g/mol
- Smiles: c12c(cc(cc2)S(=O)(=O)[O-])ccc(c1/N=N/c1c(cc(c(c1)C)S(=O)(=O)[O-])OC)O.[Na+].[Na+]
- InChI :1S/C18H16N2O8S2.2Na/c1-10-7-14(16(28-2)9-17(10)30(25,26)27)19-20-18-13-5-4-12(29(22,23)24)8-11(13)3-6-15(18)21;;/h3-9,21H, 1-2H3,(H,22,23,24)(H,25,26,27);;/q;2*+1/p-2/b20-19+;;
- Substance type: Organic
- Physical state: Red powder
- Purity: 96.6% (pure dye content)

Test animals

Species:

rat

Strain:

other: Charles River Ceesarean albino

Sex:

male/female

Details on test animals or test system and environmental conditions:

No data available

Administration / exposure

Route of administration:

oral: feed

Vehicle:

not specified

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
The test chemical mixed with feed
DIET PREPARATION
- Rate of preparation of diet (frequency): No data available
- Mixing appropriate amounts with (Type of food): No data available
- Storage temperature of food: No data available

VEHICLE
- Justification for use and choice of vehicle (if other than water):
- Concentration in vehicle: 3700, 13,900 and 51,900 ppm - Amount of vehicle (if gavage):
- Lot/batch no. (if required): No data available
- Purity: No data available

Details on mating procedure:

- M/F ratio per cage: 1:2
- Length of cohabitation: 21 days
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy days. The first day that sperm were observed was designated as 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): The females were then placed in individual cages.
- Any other deviations from standard protocol: No data available

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

27 Weeks

Frequency of treatment:

Daily

Doses / concentrations

Remarks:

0,3700, 13,900 and 51,900 ppm (equivalent to 0, 185, 695 and 2595 mg/kg bw per day)

No. of animals per sex per dose:

Total: 120
0 mg/kgbw/day: 10 male and 20 female
185mg/kgbw/day: 10 male and 20 female
695mg/kgbw/day: 10 male and 20 female
2595 mg/kgbw/day: 10 male and 20 female

Control animals:

yes, plain diet

Details on study design:

No data available

Positive control:

No data available

Examinations

Parental animals: Observations and examinations:

Clinical observations included food consumption, appearance, individual body weights and behavior and were made weekly.

Oestrous cyclicity (parental animals):

No data available

Sperm parameters (parental animals):

No data available

Litter observations:

individual fetal weight and length (crown to rump), and external fetal anatomical structure were observed

Postmortem examinations (parental animals):

SACRIFICE
- Male animals: All surviving animals [describe when, e.g. as soon as possible after the last litters in each generation were produced.]
- Maternal animals: All surviving animals [describe when, e.g. after the last litter of each generation was weaned.]

GROSS NECROPSY
-yes , Observations included number and placement of implantation sites, resorption sites, and live and dead fetuses; individual fetal weight and length (crown to rump), and external fetal anatomical structure. Gross necropsies were performed on each female including examination of uterus and visceral structures.

HISTOPATHOLOGY / ORGAN WEIGHTS
The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively.

Postmortem examinations (offspring):

SACRIFICE
- Twenty-four hours following the birth of the pups the first litters (FIA) were arbitrarily reduced to 8 maximum per mother.
- These animals were subjected to postmortem examinations (macroscopic and/or microscopic examination) as follows:

GROSS NECROPSY
- Gross necropsy consisted of [external and internal examinations including the cervical, thoracic, and abdominal viscera.]

HISTOPATHOLOGY / ORGAN WEIGTHS
The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively.

Statistics:

No data available

Reproductive indices:

No data available

Offspring viability indices:

No data available

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

not specified

Dermal irritation (if dermal study):

not specified

Mortality:

not specified

Body weight and weight changes:

not specified

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

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):

Fertility indices for the control and test animals of both F1A and F1B were considered low.The fertility index of the 3700 ppm test group in the F2A breeding cycle as well as the 3700 and 51900 ppm test groups in the F2B breeding cycle were reported to be low in comparison to control animals and historical control data.

Effect levels (P0)

Dose descriptor:

NOAEL

Effect level:

695 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

reproductive performance

Remarks on result:

other: No toxic effects were observed

Target system / organ toxicity (P0)

Critical effects observed:

not specified

System:

other: not specified

Organ:

not specified

Results: P1 (second parental generation)

General toxicity (P1)

Clinical signs:

not specified

Dermal irritation (if dermal study):

not specified

Mortality:

not specified

Body weight and weight changes:

not specified

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

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

Reproductive function / performance (P1)

Reproductive function: oestrous cycle:

not specified

Reproductive function: sperm measures:

not specified

Reproductive performance:

effects observed, treatment-related

Description (incidence and severity):

The fertility index of the 3700 ppm test group in the F2A breeding cycle as well as the 3700 and 51900 ppm test groups in the F2B breeding cycle were reported to be low in comparison to control animals and historical control data.

Effect levels (P1)

Dose descriptor:

NOAEL

Effect level:

695 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

reproductive performance

Remarks on result:

other: No toxic effects were observed

Target system / organ toxicity (P1)

Critical effects observed:

not specified

System:

other: not specified

Organ:

not specified

Results: F1 generation

General toxicity (F1)

Clinical signs:

not specified

Dermal irritation (if dermal study):

not specified

Mortality / viability:

mortality observed, treatment-related

Body weight and weight changes:

not specified

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:

not specified

Histopathological findings:

not specified

Other effects:

no effects observed

Description (incidence and severity):

Growth suppression characterized as slight was also reported for the low-level F1B pups, and the high-level F1A and F1B pups and the F2A . when compared with controls. All other measured parameters were comparable to controls in each generation and among the two filial generations.

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:

695 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No evidence was seen of teratogenic or embryotoxic effects were observed

Remarks on result:

other: No evidence was seen of teratogenic or embryotoxic effects were observed

Target system / organ toxicity (F1)

Critical effects observed:

not specified

System:

other: not specified

Organ:

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:

not specified

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:

not specified

Histopathological findings:

not specified

Other effects:

no effects observed

Description (incidence and severity):

Growth suppression characterized as slight was also reported for the F2A and F2B breeding cycles when compared with controls. All other measured parameters were comparable to controls in each generation and among the two filial generations.

Developmental neurotoxicity (F2)

Behaviour (functional findings):

not specified

Developmental immunotoxicity (F2)

Developmental immunotoxicity:

not specified

Effect levels (F2)

Dose descriptor:

NOAEL

Generation:

F2

Effect level:

695 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No evidence was seen of teratogenic or embryotoxic effects were observed

Remarks on result:

other: No evidence was seen of teratogenic or embryotoxic effects were observed

Target system / organ toxicity (F2)

Critical effects observed:

not specified

System:

other: not specified

Organ:

not specified

Overall reproductive toxicity

Reproductive effects observed:

not specified

Treatment related:

not specified

Applicant's summary and conclusion

Conclusions:

The NOAEL was considered to be 695 mg/kg bw for F0 and F1 generation , when male and female rats were treated with test chemical orally .

Executive summary:

Two generation reproductive toxicity study of test chemical was performed on male and femaleCharles River Ceesarean albino rats . The test chemical mixed with food in dose concentration 0,3700, 13,900 and 51,900 ppm  (equivalent to 0, 185, 695 and 2595 mg/kg bw per day) were administered for 27 weeks .These P1 parental generations were individually housed. Clinical observations included food consumption, appearance, individual body weights and behavior and were made weekly. The F1A weanling rats designated P2 generation were kept 4-5 to a cage according to sex and maintained on the same concentration level as their parents until reaching maturity. During the breeding phase of the P1 generation, two females and one male were placed in a breeding cage. At weekly intervals during the mating period, the males were rotated among the females in each group. Following mating, the females were placed in individual cages to produce the first (FIA) litters. Twenty-four hours following the birth of the pups the first litters (FIA) were arbitrarily reduced to 8 maximum per mother. The number of conceptions, number of litters, live births, stillbirths, size of natural and nursing litters, deaths during the period of lactation, and number of pups weaned were recorded. The body weights of each pup were recorded at 24 hours and at weaning. Gross signs of toxicity were monitored. After 21-days of nursing, random pups were sacrificed and gross necropsies performed. Twenty-four females and twelve males remaining from each test group and control group were selected at random and designated the P2 generation. Following the weaning of the F1A animals, the P1 generation was remated to produce their second litters referred to as F1B, according to the procedures described above.
The P2 generation was housed 4-5 per cage and was maintained on the same dietary levels as their parents. The procedures outlined above for the P1 generation were maintained for the P2 generation. The litters of the P2 animals were referred to as the F2A litters. Body weights of the F2A pups were monitored 24 hours following the birth and at weaning. Gross signs of toxicity were recorded. Following a 21-day nursing period, all pups were weaned and sacrificed. One week following the weaning period of the F2A litter, the P2 generation was remated to produce their second litters (F2B). Two females were placed in a cage with a male from the corresponding dose group. Males were rotated weekly, and females were examined daily for presence of spermatozoa for a maximum of 21 consecutive days. The first day that sperm were observed was designated as day 0 of gestation. The females were then placed in individual cages. Half of the females (12) were sacrificed on day 19 or 20 of gestation and Caesarean sections were performed. Observations included number and placement of implantation sites, resorption sites, and live and dead fetuses; individual fetal weight and length (crown to rump), and external fetal anatomical structure. Gross necropsies were performed on each female including examination of uterus and visceral structures. The remaining 12 females were allowed to litter normally. The fetuses of both females delivering normally and via Caesarean section were necropsied.  

 

Fertility indices for the control and test animals of both F1A and F1B were considered low. The authors attributed this to the advanced age of the animals upon mating. The fertility index of the 3700 ppm test group in the F2A breeding cycle as well as the 3700 and 51900 ppm test groups in the F2B breeding cycle were reported to be low in comparison to control animals and historical control data.
Offspring toxicity F1 and F2: Growth suppression characterized as slight was also reported for the low-level F1B pups, and the high-level F1A and F1B pups and the F2A and F2B breeding cycles when compared with controls. All other measured parameters were comparable to controls in each generation and among the two filial generations. HenceThe NOAEL was considered to be 695 mg/kg bw for F0 and F1 generation , when male and female rats were treated with test chemical orally .