reaction product of: C.I. Leuco Sulfur Black 1 with (3-chloro-2-hydroxypropyl)trimethylammonium chloride

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 23. Aug. 1995 to 06. Sep. 1995

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1995

Report date:

1995

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Version / remarks:

1987

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

1992

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

424-510-1

EC Name:

-

Cas Number:

220150-59-4

Molecular formula:

not applicable for UVCB substance

IUPAC Name:

Reaction products of Phenol, 2,4-dinitro-, sulfurized, leuco derivatives and (3-chloro-2-hydroxypropyl)trimethylammonium chloride

Test animals

Species:

rat

Strain:

Wistar

Remarks:

Hanlbm: WIST (SPF)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: BRL, Biological Research Laboratories Ltd., Wolferstrasse 4, 4414 Füllinsdorf, CH
- Females nulliparous and non-pregnant: not specified
- Age at study initiation: males: 8 weeks; females: 11 weeks
- Weight at study initiation: males: 220 to 239 g; females: 187 to 213 g
- Fasting period before study: not specified
- Housing: acclimatisation: in groups of five in Makrolon type-4 cages; during treatment and observation: individually in Makrolon type-3 cages. Autoclaved standard softwood bedding ("Lignocel", Schill AG, 4132 Muttenz, CH).
- Diet: ad libitum; pelleted standard Kliba 343, batch nos. 86/95 and 65/95 rat maintenance diet ("Kliba", Klingentalmühle AG, 4303 Kaiseraugst, CH). Results of analyses for contaminants included in report.
- Water: ad libitum; community tap water from Füllinsdorf. Bacteriological, chemical and contaminant analyses included.
- Acclimation period: one week under laboratory conditions, after health examination. Only animals without any visible signs of illness were used for the study.
- Identification: by unique cage number and corresponding colour-coded spots on the tail
- Randomisation: randomly selected at the time of delivery into groups of five

ENVIRONMENTAL CONDITIONS
- Temperature: 21.5 to 23 °C
- Humidity: 54 to 76 %
- Air changes: 10 to 15 per hour
- Photoperiod (hrs dark / hrs light): 12 / 12

Administration / exposure

Type of coverage:

semiocclusive

Vehicle:

water

Remarks:

bi-distilled water

Details on dermal exposure:

TEST SITE
- Area of exposure: dorsal area, clipped one day before test item application
- Coverage: 10 % of body surface area
- Type of wrap if used: semi-occlusive

REMOVAL OF TEST SUBSTANCE
- Washing: washed with lukewarm tap water and dried with disposable paper towels
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount applied: 4 mL/kg bw
- Concentration: 2000 mg/kg bw
- Constant volume used: no
- Constant concentration used: yes
- For solids, paste formed: yes

Duration of exposure:

24 hours

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

Males (2000 mg/kg bw): 5
Females (2000 mg/kg bw): 5
Total: 10

Control animals:

no

Details on study design:

- Observations: clinical signs, body weight, mortality, pathological change
- Duration of observation period following administration: 14 days
- Frequency of observations: four times during test day 1 then daily during days 2 to 15
- Frequency of weighing: on test days 1 (pre-administration), 8 and 15
- Necropsy of survivors performed: yes. Necropsies were performed by experienced prosectors. At the end of the observation period all animals were anaesthetised by intraperitoneal injection of NARCOREN (Rhone Merieux GmbH, 88471 Laupheim, Germany) at a dose of at least 2.0 ml/kg bw (equivalent to at least 320 mg/kg sodium pentobarbitone /kg bw) and sacrificed by exsanguination. The animals were examined macroscopically.

CLINICAL SIGNS:
General behaviour: aggressiveness, vocalisation, restlessness / excitation, nervousness, fear, sedation, somnolence, sleep, coma
Respiration: apnoea, dyspnea, rales
Eye: chromodacryorrhoea, exophthalmos, miosis, mydriasis, whitish discharge, lid adhesion, lacrimation, negative corneal reflex
Nose: rhinorrhoea, epistaxis
Motility: akinesia, ataxia, dropped head, hyperkinesia, hypokinesia, paralysis (flaccid), paralysis (spastic), paddling movements, stiff gait, rolling movements
Body posture: ventral body position, latero-abdominal position, hunched posture
Motor susceptibility: spasms, tonic muscle spasms, clonic muscle spasms, opisthotonus, saltatory spasms, trismus, tremor, muscle-twitching, muscle-twitching (generalised)
Skin: erythema, oedema, necrosis
Various: loss of weight, emaciation, diarrhoea, ruffled fur, salivation, pallor, cyanosis

Statistics:

The LOGIT-Model could not be used as no deaths occurred.

Results and discussion

Effect levels

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No deaths occurred during the study period.

Clinical signs:

other: There were no clinical signs of systemic toxiclty. The skin was discoloured with blue at the application site in all animals. The staining persisted on test days 2 to 15. In one male animal the blue discoloured skin was observed until test day 8.

Gross pathology:

No macroscopic findings were noted.

Applicant's summary and conclusion

Interpretation of results:

other: not classified according to the CLP criteria (EC 1272/2008)

Conclusions:

The LD50 (dermal, rat) of test item was found to be greater than 2000 mg/kg bw.

Executive summary:

Acute dermal toxicity of the test item was evaluated in an experimental study that was performed according to the OECD Guideline 402 (1987) and EU method B.3 (1992). 2000 mg/kg bw of the test item was applied dorsally to 10 rats of both sexes for a duration of 24 hours.

A death rate of 0 % at 2000 mg/kg bw was observed. Local discolouration was observed between 24 hours and end of observation period. The LOGIT-Model could not be applied to these data. The acute dermal toxicity of the test item in rats of both sexes observed over a period of 15 days was estimated to be greater than 2000 mg/kg bw. Therefore, it can be extrapolated that the LD50 (dermal, rat) is greater than 2000 mg/kg bw.