Reaction mass of Chromate(1-), [2-(3-chlorophenyl)-2,4-dihydro-4-[[2-hydroxy-5-(methylsulfonyl)phenyl]azo]-5-methyl-3H-pyrazol-3-onato(2-)][methyl [7-hydroxy-8-[[2-hydroxy-5-(methylsulfonyl)phenyl]azo]-1-naphthalenyl]carbamato(2-)]-, sodium and sodium bis[2-(3-chlorophenyl)-2,4-dihydro-4-[[2-hydroxy-5-mesylphenyl]azo]-5-methyl-3H-pyrazol-3-onato(2-)]chromate(1-) and sodium bis[methyl [7-hydroxy-8-[[2-hydroxy-5-mesylphenyl]azo]-1-naphthyl]carbamato(2-)]chromate(1-)

Endpoint summary

Description of key information

Oral:

In an acute oral toxicity study according to OECD Guideline 423 (acute toxicity class method) in rats (BASF SE, 2017), an LD50 between 300 and 2000 mg/kg bw was determined.

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:

2016-10-28 to 2016-12-09

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2001-12-17

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

no

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Batch No. of test material: 001-140902, Test item No.: 16/0083-1
- Expiration date of the batch: 2019-06-07

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature

OTHER SPECIFICS: Solid / brown

Species:

rat

Strain:

Wistar

Remarks:

Crl:WI (Han) SPF

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Wiga GmbH, Germany
- Females nulliparous and non-pregnant: yes
- Age at study initiation: ~ 10 weeks
- Weight at study initiation: 177 - 184 g
- Fasting period before study: Feed was withdrawn from the animals at least 16 hours before administration, but water was available ad libitum.
- Housing: Single housing in fully air-conditioned rooms, Makrolon cage, type III.
- Diet: VRF1(P); SDS Special Diets Services, 67122 Altrip, Germany, ad libitum
- Water: Tap water, ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 30 - 70
- Air changes (per hr): ~ 10
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

deionized

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 20,3 g/100mL
- Amount of vehicle: 10 mL/kg bw
- Justification for choice of vehicle: Aqueous preparation corresponds to the physiological medium

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: By request of the sponsor a starting dose of 2000 mg/kg bw was chosen in the first step with 3 female animals. As all animals died, 300 mg/kg bw were administered to 3 female rats in the second step. Because no mortality occurred, 300 mg/kg bw were administered to another group of 3 female animals in the third step.

Doses:

2000, 300 mg/kg bw

No. of animals per sex per dose:

3 female animals/dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical signs for each animal were recorded several times on the day of administration and at least once during each workday thereafter. Individual body weights shortly before administration (day 0), weekly thereafter, on the last day of observation and on the day of death starting with study day 1. A check for any dead or moribund animals was made at least once each workday
- Necropsy of survivors performed: yes, necropsy with gross pathological examination was performed on the last day of the observation period after sacrifice by CO2-inhalation in a chamber with gradually increasing concentrations. Necropsy of all animals that died were performed as early as possible after death.
- Other examinations performed: No histological examinations were performed.

Statistics:

Calculations were performed using Microsoft Excel 2010 and checked with a calculator.

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 300 - < 2 000 mg/kg bw

Based on:

test mat.

Mortality:

One animal of the single 2000 mg/kg bw.test group died on study day 2, while the two other animals showed delayed mortality on study day 6. No mortality occurred in both 300 mg/kg bw test groups.

Clinical signs:

other: Clinical signs in the single 2000 mg/kg test group revealed in the first animal red-brownish discolored feces on day 1 after administration, which persisted until day 3. Reduced defecation could be noted on study day 2. Impaired general state, piloerectio

Gross pathology:

The following macroscopic pathologic findings were observed in the animals that died (2000 mg/kg bw test group, 3 females):
- Black discoloration of the stomach contents in one animal
- Red discoloration of the glandular stomach in one animal
- Red discoloration of the small intestine, red-brownish discolored contents in one animal
Due to the advanced putrefaction no macroscopic pathological findings could be determined in the other two animals which died in day 6.
There were no macroscopic pathological findings in the animals sacrificed at the end of the observation period (both 300 mg/kg bw. test groups, 6 females).

Interpretation of results:

Category 4 based on GHS criteria

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

300 mg/kg bw

Quality of whole database:

Klimisch code 1

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Oral:

In an acute oral toxicity study performed according to the Acute Toxic Class Method (OECD Guideline 423), doses of 2000 or 300 mg/kg bw of the undiluted test substance were administered by gavage to test groups of three fasted Wistar rats each (3 females). All three animals of the 2000 mg/kg bw dose group died within 6 days. Animals showed an impaired general state, piloerection, diarrhea, red-brownish discolored feces. Pathological findings included black discoloration of the stomach contents, red discoloration of the glandular stomach, red discoloration of the small intestine. Due to the advanced putrefaction no macroscopic pathological findings could be determined in the other two animals which died on day 6.

No mortality occurred in both 300 mg/kg bw test groups. In both groups, animals showed an impaired general state and piloerection. The body weight of all surviving animals in both 300 mg/kg bw test groups increased throughout the study period within the normal range. There were no macroscopic pathological findings in the animals sacrificed at the end of the observation period (both 300 mg/kg bw test groups, 6 females).

The acute oral LD50 was determined to be above 300 and below 2000 mg/kg bw.

Justification for classification or non-classification

The available experimental test data are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008.

As a result the substance is considered to be classified for acute oral toxicity (Category 4, H302) under Regulation (EC) No 1272/2008, as amended for the tenth time in Regulation (EU) No 2017/776.