Glycine, N-[5-(acetylamino)-4-[2-(2-bromo-4,6-dinitrophenyl)diazenyl]-2-methoxyphenyl]-N-(2-methoxy-2-oxoethyl)-, methyl ester

Endpoint summary

Description of key information

Disperse Blue ANT, did not induce toxic effects in the rat following oral administration of a single dose at a level of 2000 mg/kg body weight.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 02 December 2014 to 02 January 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Compliant to GLP and testing guideline, coherence between data, results and conclusions.

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Deviations:

no

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Italy s.r.l., San Pietro al Natisone (UD), Italy
- Age at study initiation: approximately 9 - 10 weeks
- Weight at study initiation: 162-170 grams
- Fasting period before study: approximately 16 hours prior to dosing
- Housing: 3 animals/cage
- Diet (e.g. ad libitum): ad libitum, except for the fasting period prior to dosing
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 °C+/-2 °C
- Humidity (%): 55%+/-15%
- Air changes (per hr): Approximately 15 to 20 air changes per hour
- Photoperiod (hrs dark / hrs light): Artificial (fluorescent tubes), daily light/dark cycle of 12/12 hours

IN-LIFE DATES: From: To: From: 09 December 2014 (Allocation of the animals to the first group) To: 02 January 2015 (Last necropsy procedure)

Route of administration:

oral: gavage

Vehicle:

polyethylene glycol

Remarks:

: water 1:1

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 200 mg/mL
- Amount of vehicle (if gavage): 10 mL/kg body weight
- Justification for choice of vehicle: solubility of the test item


MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg body weight

DOSAGE PREPARATION (if unusual):

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose:

Doses:

2000 mg/kg body weight

No. of animals per sex per dose:

3 females

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: All animals were observed for clinical signs on dosing, approximately 0.5,
2, 4 hours after dosing on the day of treatment and daily thereafter. All animals were weighed at allocation to the
study (Day -1), on the day of dosing (Day 1) and on Days 2, 8 and 15.
- Necropsy of survivors performed: yes, gross necropsy examination for both external and internal abnormalities,
with particular attention to the gastro-intestinal tract.

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred.

Clinical signs:

other: In the first group of animals, blue staining on the dorsal region and perianal region was individually observed from Day 2 up to Day 8. In the second group of 3 female animals, generalised blue staining (i.e. on the dorsum/dorsal region, hindlimb/forelim

Gross pathology:

No abnormalities were observed at necropsy examination.

Interpretation of results:

study cannot be used for classification

Remarks:

Migrated information

Conclusions:

These results indicate that the test item, Disperse Blue ANT, did not induce toxic effects in the rat following oral administration of a single dose
at a level of 2000 mg/kg body weight. The lack of mortality demonstrates the acute toxicity expected (ATE) to be greater than 2000 mg/kg body weight.

Executive summary:

The acute toxicity of Disperse Blue ANT was investigated following a single oral administration to the Sprague Dawley rat followed by a 14-day observation period. A first group of 3 female animals was initially dosed at 2000 mg/kg body weight (Step 1). No mortality occurred and no signs of toxicity were noted. The only finding observed was blue staining on perianal and/or sacral dorsal region in all animals from Day 2 up to Day 8. A second group of 3 female animals was then dosed at the same dose level (Step 2). No death occurred and no signs of toxicity were seen. Generalised blue staining was individually observed starting from the day of dosing up to Day 7. Body weight changes recorded during the study were within the expected range for this strain and age of animals. No abnormalities were observed at necropsy examination performed at the end of the observation period on animals of both groups. These results indicate that the test item, Disperse Blue ANT, did not induce toxic effects in the rat following oral administration of a single dose at a level of 2000 mg/kg body weight.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 02 December 2014 to 18 December 2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Compliant to GLP and testing guideline, coherence between data, results and conclusions.

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

yes

Remarks:

At the end of the exposure period (23.5 hours and not 24 as indicated in protocol), the treatment area was cleaned with corn oil and not with lukewarm water. These are deviations did not compromise the integrity of the study.

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Italy s.r.l., San Pietro al Natisone (UD), Italy
- Age at study initiation: approximately 8-10 weeks (at allocation)
- Weight at study initiation: 174-182 grams
- Housing: individually caged during the study
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at lest 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 °C+/-2 °C
- Humidity (%): 55%+/-15%
- Air changes (per hr): Approximately 15 to 20 air changes per hour
- Photoperiod (hrs dark / hrs light): Artificial (fluorescent tubes), daily light/dark cycle of 12/12 hours

IN-LIFE DATES: From: To: From: 03 December 2014 (allocation day) To: 18 December2014 (day of necropsy)

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: darsal surfaces of the trunk
- % coverage: approximately 10% of body surface
- Type of wrap if used: a patch of surgical gauze covered by a strip of synthetic film was placed over the treated site and the whole assembly held in place byencircling the trunk of the animal with a lenght of elastic adhesive bendage.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): corn oil
- Time after start of exposure: approximately 24 hours (23.5 hours)

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg/body weight
- For solids, paste formed: yes, using 1 mL of sterile water

Duration of exposure:

Approximately 24 hours (23.5 hours)

Doses:

2000 mg/kg/body weight

No. of animals per sex per dose:

5 males and 5 females

Control animals:

not required

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: All animals were observed for clinical signs on dosing, approximately 1, 2, 4 hours hours after dosing on the day
of treatmnent and daily thereafter. All animals were weighed at allocation to the study (Day -1), on the day of dosing (Day 1) and on Days 8 and 15.
- Necropsy of survivors performed: yes, gross necropsy examination for both external and internal abnormalities, with particular attention to the treatment
site.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred.

Clinical signs:

other: Blue and/or yellow staining of the treated site was observed in all animals from Day 2 up to Day 15.

Gross pathology:

Yellow staining of the treatment site was observed during the external examination performed on all animals at termination of the study. In two
single occasions (animal nos. A0674004 and A0674005), also blue staining was noted. No internal abnormalities were seen at necropsy examination.

Interpretation of results:

study cannot be used for classification

Remarks:

Migrated information

Conclusions:

These results indicate that the test item, Disperse Blue ANT, has no toxic effect on the rat following dermal exposure over a 24 hour period at a
level of 2000 mg/kg body weight. The lack of mortality demonstrates the LD50 to be greater than 2000 mg/kg body weight.

Executive summary:

The acute toxicity of Disperse Blue ANT was investigated following dermal administration of a single dose to the rat followed by a 14-day observation period. A single dose of 2000 mg/kg body weight was administered to a group of 5 male and 5 female animals for 24 hours. After 14 days all animals were killed and subjected to necropsy examination. No mortality occurred and no signs of toxicity were observed in male or female animals during the observation period. Staining of the treatment site was seen during the entire observation period of the study starting from the day after dosing. The body weight changes observed during the study were within the expected range for this species and age of animals. Staining of the treatment site was found in all animals at termination of the study. No internal abnormalities were seen. These results indicate that the test item, Disperse Blue ANT, has no toxic effect on the rat following dermal exposure over a 24 hour period at a level of 2000 mg/kg body weight. The lack of mortality demonstrates the LD50 to be greater than 2000 mg/kg body weight.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Additional information

The acute toxicity of Disperse Blue ANT was investigated following a single oral administration to the Sprague Dawley rat followed by a 14-day observation period. A first group of 3 female animals was initially dosed at 2000 mg/kg body weight (Step 1). No mortality occurred and no signs of toxicity were noted. The only finding observed was blue staining on perianal and/or sacral dorsal region in all animals from Day 2 up to Day 8. A second group of 3 female animals was then dosed at the same dose level (Step 2). No death occurred and no signs of toxicity were seen. Generalised blue staining was individually observed starting from the day of dosing up to Day 7. Body weight changes recorded during the study were within the expected range for this strain and age of animals. No abnormalities were observed at necropsy examination performed at the end of the observation period on animals of both groups. These results indicate that the test item, Disperse Blue ANT, did not induce toxic effects in the rat following oral administration of a single dose at a level of 2000 mg/kg body weight.

Justification for classification or non-classification

Classification:        No category

Signal word:        No signal word required

Hazard statement : No hazard statement required