Phenol, 4-(phenylamino)-, sulfurized, leuco deriv.

Endpoint summary

Description of key information

Acute Toxicity: oral:

In an acute oral toxicity study with female rats, performed according to OECD 423 guidelines, an LD50 of >2000 mg/kg bw was determined.

 

Dermal:

In an acute dermal toxicity study with rats, performed according to OECD 402 guidelines, an LD50 of >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:

02 - 22 November 2012

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)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nohsan, Notification No 8147, April 2011, including the most recent partial revisions.

Deviations:

no

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

- Source: Charles River Deutschland, Sulzfeld, Germany.
- Age at study initiation: Young adult animals (approx. 8-9 weeks old)
- Weight at study initiation: Body weight variation was within +/- 20% of the sex mean (first group: 169 - 188 grams; second group: 159-167 grams).
- Fasting period before study: Animals were deprived of food overnight prior to dosing and until 3-4 hours after administration of the test substance. Water was available ad libitum.
- Housing: Group housing of 3 animals per cage in labeled Macrolon cages
- Diet: Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).
- Water: Free access to tap water.
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
Environmental controls for the animal room were set to maintain 18 to 24°C, a relative humidity of 40 to 70%, approximately 15 room air changes/hour, and a 12-hour light/12-hour dark cycle.

IN-LIFE DATES: From: 02 to 22 November 2012

Route of administration:

oral: gavage

Vehicle:

CMC (carboxymethyl cellulose)

Details on oral exposure:

GAVAGE METHOD: plastic feeding tubes.

Frequency:
Two dosages within 24 hours. The first on t=0 and the second on t=4 hours. Multiple dosages given within 24 hours are regarded as a single dose (see also dose volume).

The maximum time between formulation and the second dosing on each day was exceeded with a maximum of approximately 0.75 hours.

Fasting:
Animals were deprived of food overnight prior to dosing and until approximately 2 hours after the second administration of the test substance. Water was available ad libitum.

The maximum time of fasting of 24 hours was exceeded with 2 minutes for the second group at 2000 mg/kg.

VEHICLE
- Justification for choice of vehicle: The vehicle was selected based on trial formulations performed at WIL Research Europe and on test substance data supplied by the sponsor.

MAXIMUM DOSE VOLUME APPLIED: 2000 mg/kg (10 mL/kg) body weight.

DOSAGE PREPARATION:
The formulations (w/w) were prepared within 5 hours or within approximately 5.75 hours prior to the first and second dosing on each day, respectively. Homogeneity was accomplished to a visually acceptable level. Correction was made for purity of the test substance (16.4%).

Doses:

2000 mg/kg body weight

No. of animals per sex per dose:

6 (2 groups of three females in a stepwise manner)

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Mortality/Viability: Twice daily
Body weights: Days 1 (pre-administration), 8 and 15.
Clinical signs: At periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15.
- Necropsy of survivors performed: At the end of the observation period, all animals were sacrificed by oxygen/carbon dioxide procedure and subjected to necropsy. Descriptions of all internal macroscopic abnormalities were recorded.
- Other examinations performed: none.

Statistics:

No statistical analysis was performed (The method used is not intended to allow the calculation of a precise LD50 value).

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred.

Clinical signs:

Hunched posture was shown by three animals on Day 1.

Black faeces noted for both groups at 2000 mg/kg between Days 2 and 4, was considered to be related to staining properties of the test substance (a black paste).

Body weight:

The body weight gain shown by the animals over the study period was considered to be similar to that expected of normal untreated animals of the same age and strain.

Gross pathology:

No abnormalities were found at macroscopic post mortem examination of the animals.

Interpretation of results:

GHS criteria not met

Conclusions:

In an acute oral toxicity study with female rats, performed according to OECD 423 guidelines, an LD50 of >2000 mg/kg bw was determined.

Executive summary:

In an acute oral toxicity study, the test substance was administered by oral gavage to two subsequent groups of three female Wistar rats at 2000 mg/kg body weight. The total dose was administered as two dosages of 1000 mg/kg body weight each (the first on t=0 and the second on t=4 hours), due to the low purity of the test substance. Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice (Day 15).

Hunched posture was shown by three animals on Day 1. Black faeces noted for both groups at 2000 mg/kg between Days 2 and 4, was considered to be related to staining properties of the test substance (a black paste). The body weight gain shown by the animals over the study period was considered to be normal. No mortality occurred and no abnormalities were found at macroscopic post mortem examination of the animals.

The oral LD50 value in Wistar rats was established to exceed 2000 mg/kg body weight.

According to the OECD 423 test guideline, the LD50 cut-off value was considered to exceed 5000 mg/kg body weight.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

2 000 mg/kg bw

Quality of whole database:

Reliable guideline study data

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:

06-20 December 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nohsan, Notification No 8147, April 2011; including the most recent partial revisions.

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

- Source: Charles River Deutschland, Sulzfeld, Germany.
- Age at study initiation: Young adult animals (approx. 10 weeks old)
- Weight at study initiation: Body weight variation was within +/- 20% of the sex mean (males: 281 - 307 grams; females: 191 - 204 grams).
- Housing: Individual housing in labeled Macrolon cages
- Diet: Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).
- Water: Free access to tap water.
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
Environmental controls for the animal room were set to maintain 18 to 24°C, a relative humidity of 40 to 70%, approximately 15 room air changes/hour, and a 12-hour light/12-hour dark cycle.

IN-LIFE DATES: From: 06 to 20 December 2012

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

One day before exposure (Day -1) an area of approximately 5x7 cm on the back of the animal was clipped.

The formulation was applied in an area of approx. 10% of the total body surface, i.e. approx. 25 cm² for males and 18 cm² for females. The formulation was held in contact with the skin with a dressing.

Frequency: Single dosage, on Day 1.

Washing: Following application, dressings were removed and the skin cleaned of residual test substance using tap water.

Duration of exposure:

24 hours.

Doses:

2000 mg/kg

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

Dose volume: 2000 mg/kg (based on main component C.I. Leuco Sulphur Brown 96). This dose corresponded to 10200 mg test substance including C.I. Leuco Sulphur Brown 96, based on a correction factor of 5.1 (to correct for water content of the test substance).

DOSAGE PREPARATION: The test substance was dosed undiluted as delivered by the sponsor. Correction was made for
water content of the test substance (correction factor 5.1).

Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Mortality/Viability: Twice daily.
Body weights: Days 1 (pre-administration), 8 and 15.
Clinical signs: At periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15.
- Necropsy of survivors performed: At the end of the observation period, all animals were sacrificed by oxygen/carbon dioxide procedure and subjected to necropsy. Descriptions of all internal macroscopic abnormalities were recorded.
- Other examinations performed: none.

Statistics:

None.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred.

Clinical signs:

Flat/hunched posture, piloerection, ptosis, uncoordinated movements, hypothermia and/or chromodacryorrhoea were noted among all animals on Days 1 and/or 2.

The treated skin area of all animals showed black staining on Days 2 and 3 and red staining between Days 4 and 15. Other findings that were noted on the treated skin area included swelling, focal erythema and scales between Days 2 and 5.

Body weight:

The changes noted in body weight gain were within the range expected for rats used in this type of study and were therefore considered not indicative of toxicity.

Gross pathology:

No abnormalities were found at macroscopic post mortem examination of the animals.

Interpretation of results:

not classified

Conclusions:

In an acute dermal toxicity study with rats, performed sccording to OECD 402 guidelines, an LD50 of >2000 mg/kg bw was determined.

Executive summary:

In an acute dermal toxicity study, the test substance was administered to five Wistar rats of each sex by a single dermal application at 2000 mg/kg body weight for 24 hours. Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice (Day 15).

No mortality occurred. Flat/hunched posture, piloerection, ptosis, uncoordinated movements, hypothermia and/or chromodacryorrhoea were noted among all animals on Days 1 and/or 2. The treated skin area of all animals showed black staining on Days 2 and 3 and red staining between Days 4 and 15. Other findings that were noted on the treated skin area included swelling, focal erythema and scales between Days 2 and 5.

The changes noted in body weight gain were within the range expected for rats used in this type of study and were therefore considered not indicative of toxicity. No abnormalities were found at macroscopic post mortem examination of the animals.

The dermal LD50 value of C.I. Leuco Sulphur Brown 96 in Wistar rats was established to exceed 2000 mg/kg body weight.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

2 000 mg/kg bw

Quality of whole database:

Reliable guideline study data

Additional information

Acute toxicity: oral

In an acute oral toxicity study, the test substance was administered by oral gavage to two subsequent groups of three female Wistar rats at 2000 mg/kg body weight. The total dose was administered as two dosages of 1000 mg/kg body weight each (the first on t=0 and the second on t=4 hours), due to the low purity of the test substance. Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice (Day 15).

Hunched posture was shown by three animals on Day 1. Black faeces noted for both groups at 2000 mg/kg between Days 2 and 4, was considered to be related to staining properties of the test substance (a black paste). The body weight gain shown by the animals over the study period was considered to be normal. No mortality occurred and no abnormalities were found at macroscopic post mortem examination of the animals.

The oral LD50 value in Wistar rats was established to exceed 2000 mg/kg body weight.

Acute toxicity: dermal

In an acute dermal toxicity study, the test substance was administered to five Wistar rats of each sex by a single dermal application at 2000 mg/kg body weight for 24 hours. Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice (Day 15).

No mortality occurred. Flat/hunched posture, piloerection, ptosis, uncoordinated movements, hypothermia and/or chromodacryorrhoea were noted among all animals on Days 1 and/or 2. The treated skin area of all animals showed black staining on Days 2 and 3 and red staining between Days 4 and 15. Other findings that were noted on the treated skin area included swelling, focal erythema and scales between Days 2 and 5.

The changes noted in body weight gain were within the range expected for rats used in this type of study and were therefore considered not indicative of toxicity. No abnormalities were found at macroscopic post mortem examination of the animals.

The dermal LD50 value of C.I. Leuco Sulphur Brown 96 in Wistar rats was established to exceed 2000 mg/kg body weight.

Justification for classification or non-classification

Classification, Labelling, and Packaging Regulation (EC) No 1272/2008

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008. Based on this data, the substanceis considered not to be classified for acute oral or acute dermal toxicity under Regulation (EC) No 1272/2008, as amended for the tenth time in Regulation (EC) No 2017/776.