Phenol, 2,4-dinitro-, sulfurized, leuco derivatives

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2009-01-20 till 2009-02-10

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline-conform study under GLP without deviations

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Report date:

2009

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

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:

Phenol, 2,4-dinitro-, sulfurized, leuco derivatives

EC Number:

266-273-5

EC Name:

Phenol, 2,4-dinitro-, sulfurized, leuco derivatives

Cas Number:

66241-11-0

Molecular formula:

Molecular formula is not available.

IUPAC Name:

Reaction products of 2,4-dinitrophenol with polysulfide, leuco derivatives, sodium salt

Details on test material:

- Name of test material : Leuco Sulfur Black 1
- Physical state: thixotrophic, black, wet paste
- Composition: Leuco Sulfur Black 1:52.75 % (w/w) (calculated by amount of by-compounds)
2,4-Dinitrochlorobenzene: < 20 ppm
2,4-Dinitrophenole: < 20 ppm
Sulfate: 970 ppm
Chloride: 750 ppm
Sodium: 1.3 % (w/w)
Water: 45.75 % (w/w) by Karl-Fischer titration
- Lot/batch No.: ESA0007595
- Expiration date of the lot/batch: 2011-07-11
- Stability under test conditions: Stable under storage conditions.
- Storage condition of test material: At room temperature (range of 20 ± 5 °C), light protected.

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Laboratories Ltd., Füllinsdorf / Switzerland
- Age at study initiation: Males: 9 weeks; females: 11 weeks
- Weight at study initiation: 182.9 g – 267.1 g
- Fasting period before study: no data
- Housing: Individually in Makrolon type-3 cages with standard softwood bedding
- Diet (e.g. ad libitum): Pelleted standard Provimi Kliba 3433 rat/mouse maintenance diet, ad libidum
- Water (e.g. ad libitum): Community tap water from Füllinsdorf ad libitum
- Acclimation period: 2009-01-20 to 2009-01-26


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 30 - 70%
- Air changes (per hr): 10 - 15
- Photoperiod (hrs dark / hrs light): 12 / 12

Administration / exposure

Type of coverage:

semiocclusive

Vehicle:

water

Details on dermal exposure:

TEST SITE
- Area of exposure: back
- % coverage: 10 % of the total body surface
- Type of wrap if used: not specified


REMOVAL OF TEST SUBSTANCE
- Washing (if done): flushed with lukewarm tap water and dapped off with disposable paper towels.
- Time after start of exposure: 24 hrs


TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2319mg/kg body weight active ingredient [corresponding to 4396 mg/kg of the test item; based on the amount of 52.75% (w/w) Leuco Sulfur Black 1] or 6 mL application volume/kg body weight
- Concentration (if solution): 0.73 g/mL
- Constant volume or concentration used: yes
- For solids, paste formed: yes


VEHICLE
- Purity: purified water

Duration of exposure:

24 hrs

Doses:

Males and Females: 2319 mg/kg bw active ingredient (corresponding to 4396 mg/kg of the test item; based on the amount of 52.75% (w/w) Leuco Sulfur Black 1)


Nota: The dose levels are in terms of the active ingredient (Leuco Sulfer Black 1; 52.75 % w/w) in the test item.

No. of animals per sex per dose:

5 males, 5 females

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Viability / Mortality: Daily during the acclimatization period, during the first 30 minutes and at approximately 1, 2, 3 and 5 hours after administration on test day 1 and twice daily during days 2-15.
Body Weights: On test days 1 (prior to administration), 8 and 15.
- Necropsy of survivors performed: yes
- Other examinations performed:
clinical signs (Daily during the acclimatization period, during the first 30 minutes and at approximately 1, 2, 3 and 5 hours after administration on test day 1. Once daily during days 2-15.)
Local Dermal Signs: Once daily during days 2 (following dressing removal) through day 15

Statistics:

No statistical analysis was used.

Results and discussion

Effect levels

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

act. ingr.

Mortality:

No deaths occurred during the study.

Clinical signs:

other: No clinical signs were observed during the course of the study.

Gross pathology:

No macroscopic findings were recorded at necropsy.

Other findings:

Local Dermal Signs
Due to moderate black staining of the treated skin after the removal of the application patch, the assessment of erythema was first prevented on test day 2 in all animals. From test day 3 to test day 7 at the latest, a slight black staining was still present. Erythema was assessable, but not noted in all animals from test day 3 to the end of the study. No oedema was observed throughout the whole oberservation period. Scaling was observed in one male on test day 4.

Any other information on results incl. tables

Table 2: Number of animals dead [and with evident toxicity] [and time range within which mortality occurred]

 

Dose (mg/kg bw)	Conc. in vehicle (%)*	Mortality (# dead/total)			Time range of deaths (hours)	Number with evident toxicity (#/total)
		Male	Female	Combined		Male	Female	Combined
Control	Control	n.a.	n.a.	n.a..	n.a..	n.a..	n.a..	n.a..
2319*	0.73 g/mL	0/5	-	n.a.	n.a.	0/5	-	n.a.
2319*	0.73 g/mL	-	0/5	n.a.	n.a.	-	0/5	n.a.

n.a.: not applicable

* 2319 mg/kg active ingredient (corresponding to 4396 mg/kg of the test item; based on the amount of 52.75% (w/w) Leuco Sulfur Black 1)

 

Table 3: Clinical/local signs (Males)

Dose mg/kg bw	Ani-mal No.	Sex	Signs	Test days
				1					2	3	4	5	6	7	8	9	10	11	12	13	14	15
				0.5*	1*	2*	3*	5*
2319^a	1	M	No clinical signs	√	√	√	√	√	√	√	√	√	√	√	√	√	√	√	√	√	√	√
			Scaling								√
			Black staining						2	1	1	1	1	1
			Erythema	§	§	§	§	§	#	0	0	0	0	0	0	0	0	0	0	0	0	0
			Oedema	§	§	§	§	§	0	0	0	0	0	0	0	0	0	0	0	0	0	0
			Eschars	§	§	§	§	§	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	2	M	No clinical signs	√	√	√	√	√	√	√	√	√	√	√	√	√	√	√	√	√	√	√
			Black staining						2	1	1	1
			Erythema	§	§	§	§	§	#	0	0	0	0	0	0	0	0	0	0	0	0	0
			Oedema	§	§	§	§	§	0	0	0	0	0	0	0	0	0	0	0	0	0	0
			Eschars	§	§	§	§	§	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	3	M	No clinical signs	√	√	√	√	√	√	√	√	√	√	√	√	√	√	√	√	√	√	√
			Black staining						2	1	1	1	1	1
			Erythema	§	§	§	§	§	#	0	0	0	0	0	0	0	0	0	0	0	0	0
			Oedema	§	§	§	§	§	0	0	0	0	0	0	0	0	0	0	0	0	0	0
			Eschars	§	§	§	§	§	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	4	M	No clinical signs	√	√	√	√	√	√	√	√	√	√	√	√	√	√	√	√	√	√	√
			Black staining						2	1	1	1	1	1
			Erythema	§	§	§	§	§	#	0	0	0	0	0	0	0	0	0	0	0	0	0
			Oedema	§	§	§	§	§	0	0	0	0	0	0	0	0	0	0	0	0	0	0
			Eschars	§	§	§	§	§	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	5	M	No clinical signs	√	√	√	√	√	√	√	√	√	√	√	√	√	√	√	√	√	√	√
			Black staining						2	1	1	1	1	1
			Erythema	§	§	§	§	§	#	0	0	0	0	0	0	0	0	0	0	0	0	0
			Oedema	§	§	§	§	§	0	0	0	0	0	0	0	0	0	0	0	0	0	0
			Eschars	§	§	§	§	§	0	0	0	0	0	0	0	0	0	0	0	0	0	0

Key: 0 no findings, 1 slight, 2 moderate,√noted

* Examinations were performed within the first 30 minutes and 1, 2, 3 and 5 hours after treatment.

§Assessment of dermal findings was not possible underneath the application patch

#Assessment of erythema was not possible due to black staining of the skin

^a2319 mg/kg active ingredient (corresponding to 4396 mg/kg of the test item; based on the amount of 52.75% (w/w) Leuco Sulfur Black 1)

no clinical signs were evident in any animal during the acclimatization period.

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The median lethal dose LD50 (rat) of Leuco Sulfur Black 1 after single dermal administration to rats of both sexes, observed over a period of 14 days is greater than 2000 mg/kg body weight

Executive summary:

The acute toxicity by dermal application of LEuco Sulfur Black 1 was investigated according to OECD Guideline 402 under GLP.

Five male and five female HanRcc:WIST (SPF) rats were treated with Leuco Sulfur Black 1 at 2319 mg active ingredient/kg (corresponding to 4396 mg test item/kg) by dermal application. The test item was formulated in purified water according to the content of the active ingredient at a concentration of 0.73 g/mL and administered at a volume dosage of 6 mL/kg. The application period was 24 hours.

No deaths occurred during the study. No clinical signs were observed during the course of the study.

Due to moderate black staining of the treated skin after the removal of the application patch, the assessment of erythema was first prevented on test day 2 in all animals. From test day 3 to test day 7 at the latest, a slight black staining was still present. Erythema was assessable, but not noted in all animals from test day 3 to the end of the study. No oedema was observed throughout the whole oberservation period. Scaling was observed in one male on test day 4.

The body weight of the animals was within the range commonly recorded for this strain and age. No macroscopic findings were observed at necropsy.