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

Acute Toxicity: other routes

Administrative data

Endpoint:

acute toxicity: other routes

Remarks:

subcutaneous

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 30. Aug. 1995 to 11. Oct. 1995

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Remarks:

No CLP classification available for subcutaneous administration data

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1995

Report date:

1995

Materials and methods

Test guideline

Test guideline 1

Qualifier:

equivalent or similar to guideline

Guideline:

other: OECD Guideline 402 (Acute Dermal Toxicity)

Version / remarks:

1987

Deviations:

yes

Remarks:

Test animals were typically below minimum weight requirement (200 g). Animals were group caged, instead of individually.

Test guideline 2

Qualifier:

equivalent or similar to guideline

Guideline:

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

Version / remarks:

1992

Deviations:

yes

Remarks:

Test animals were typically below minimum weight requirement (200 g). Animals were group caged, instead of individually.

GLP compliance:

yes (incl. QA statement)

Limit test:

no

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:

Wistar HanIbm: WIST (SPF)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: BRL, Biological Research Laboratories, Ltd., Wölferstrasse 4, 4414 Füllinsdorf, CH
- Age at study initiation: males: 7 to 8 weeks; females: 9 to 10 weeks
- Weight at study initiation: males: 184.9 to 237.3 g; females: 178.8 to 200.1 g
- Fasting period before study: not specified
- Housing: groups of five in Makrolon type-3 cages with standard autoclaved softwood bedding ("Lignocel", Schill AG, 4132 Muttenz, CH)
- Diet: ad libitum; pelleted standard Kliba 343, batch no. 65/95 rat maintenance diet ("Kliba", Klingentalmühle AG,.4303 Kaiseraugst, CH) (analytical test report demonstrates suitability)
- Water: ad libitum; community tap water from Füllinsdorf (bacteriological assay, contaminant and chemical water analysis demonstrated suitability)
- Acclimation period: one week after bill of good health
- Identification: by unique cage number and corresponding colour-coded spots on the tail
- Randomisation: randomly selected at time of delivery into groups of five

ENVIRONMENTAL CONDITIONS
- Temperature: 20 to 23 °C
- Humidity: 42 to 79 %
- Air changes: 10 to 15 per hour
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

subcutaneous

Vehicle:

water

Remarks:

bi-distilled water

Details on exposure:

TEST ITEM PREPARATION
- The test item was placed into a glass beaker on a tared Mettler PM 480 balance, and the vehicle was added. A weight by volume dilution was preparad using a homogeniser.
- Homogeneity of the test item in the vehicle was maintained during treatment using a magnetic stirrer. The preparation was made immediately prior to each dosing.

TEST MATERIAL
- Volume administered: 6 mL/kg bw
- Concentration: 20, 100 and 2000 mg/kg bw
- For solids, paste formed: no; diluted in vehicle to form liquid

DURATION
- One single dose; not removed

Doses:

Group 1: 20 mg/kg bw
Group 2: 100 mg/kg bw
Group 3: 2000 mg/kg bw

No. of animals per sex per dose:

5 males and 5 females for each dose

Control animals:

no

Details on study design:

- Observations: mortality, clinical signs, body weight, macroscopic findings
- Duration of observation period following administration: 14 days
- Frequency of observations: four times during day 1, and once daily for surviving animals during days 2-15
- Frequency of weighing: on test day 1 (pre-administration), 8 and 15 for surviving animals
- Necropsy of survivors performed: yes

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 (COX, Analysis of Binary Data, London 1977) was applied to estimate the toxicity value. Additionally, the 90%, 95% and 99% confidence limits for the toxicity for each sex and the slope of the dose response line were estimated.

Results and discussion

Effect levels

Effect levels 1

Sex:

male/female

Dose descriptor:

LD50

Effect level:

187.18 mg/kg bw

Based on:

test mat.

95% CL:

>= 86.86 - <= 501.5

Effect levels 2

Sex:

male

Dose descriptor:

LD50

Effect level:

119.01 mg/kg bw

Based on:

test mat.

95% CL:

>= 17.85 - <= 553.91

Effect levels 3

Sex:

female

Dose descriptor:

LD50

Effect level:

325.67 mg/kg bw

Based on:

test mat.

95% CL:

>= 93.35 - <= 2 937.45

Mortality:

Group 1 (20 mg/kg): 0 %
Group 2 (100 mg/kg): 30 % (3 males)
Group 3 (2000 mg/kg): l00 % (5 males, 5 females)

Clinical signs:

Group 1 (20 mg/kg): no clinical signs noted
Group 2 (100 mg/kg): sedatlon (all), convulsions (all), ventral recumbency (all), uncoordinated movements (3 males, 5 females), dyspnea (4 males, 5 females), ruffled fur (2 males)
Group 3 (2000 mg/kg): sedation (all), convulsions (all), ventral recumbency (all), dyspnea (all)

Body weight:

The rate of body weight gain of the animals during the observation period was within the normal range for rats of this strain and age

Gross pathology:

No organ abnormalities were observed at necropsy.

Applicant's summary and conclusion

Conclusions:

The LD50 (subcutaneous, rat both sexes) of the test item was determined to be 187.18 mg/kg bw.

Executive summary:

Acute dermal toxicity by subcutaneous injection of the test item was evaluated in an experimental study that was performed in a similar way to the OECD Guideline 402 (1987) and EU method B.2 (1992). The test item was administered subcutaneously at doses 20, 100 and 2000 mg/kg bw by subcutaneous injection (5 male and 5 female rats per dose). The following mortality rates were observed:

20 mg/kg bw: 0 %

100 mg/kg bw: 30 % (3 males)

2000 mg/kg bw: 100 % (5 males, 5 females)

No clinical signs were observed among test subjects administered 20 mg/kg bw. Test subjects administered either 100 or 2000 mg/kg bw test item demonstrated sedation, convulsions, ventral recumbancy and dyspnea. Test subjects administered 100 mg/kg bw also demonstrated uncoordinated movements and ruffled fur. The body weight gain was withln the normal range in animals of this strain and age.

No macroscopical findings were observed in any of the animals. Based on these observations, the mean lethal dose (LDGIT-model) for the acute subcutaneous toxicity in rats of both sexes observed for a period of 14 days is 187.18 mg/kg (for males 119.01 mg/kg; for females 325.67 mg/kg).