Phenothiazine

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2010-03-17 to 2010-04-30

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Well performed GLP study according to recent OECD guidelines.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Certificate attached to full study report.

Test type:

standard acute method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Phenothiazine

EC Number:

202-196-5

EC Name:

Phenothiazine

Cas Number:

92-84-2

Molecular formula:

C12H9NS

IUPAC Name:

10H-phenothiazine

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River, 97633 Sulzfeld, Germany
- Age at study initiation: females: 18-19 weeks; males: 8-9 weeks
- Weight at study initiation: femailes: 215-228g; males: 240-260g
- Housing: Full barrier in an air-conditioned room, animals were kept individually in IVC cages, type III H, polysulphone cages on Altomin saw fibre bedding
- Diet (e.g. ad libitum): Altromin 1324, ad libitum
- Water (e.g. ad libitum): tap water, ad libitum
- Certificates of food, water and bedding are filed at the test facility
- Acclimation period: at least five days
- Female rats were nulliparous and non-pregnant

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22°C +/- 3°C
- Humidity (%): 55% +/- 10%
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12hrs / 12hrs

Administration / exposure

Type of coverage:

semiocclusive

Vehicle:

other: Corn oil (Sigma-Aldrich)

Details on dermal exposure:

TEST SITE
- Area of exposure: dorsal area of the trunk (approx. 24hrs before the test, the fur was removed by using an electrical clipper)
- % coverage: 10%
- Type of wrap if used: semi-occlusive dressing consisted of a gauze-dressing and non-irritating tape and was fixed with an additional dressing in a suitable manner.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): at the end of exposure period residual test item was removed by using corn oil
- Time after start of exposure: 24hrs

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): at a single dose of 2000 mg/kg body weight
- Constant volume or concentration used: yes
- For solids, paste formed: yes, moistened with vehicle
- The test item was ground to a fine powder in a mortar with the help of a pestle
- In order to ensure good skin contact it was moistened with the vehicle

VEHICLE
- Lot/batch no. (if required): lot no. 128K0040, expiry date: 25/04/2010
- Rationale: This vehicle was chosen due to its non-irritating characteristics

Duration of exposure:

24hrs

Doses:

2000mg/kg body weight

No. of animals per sex per dose:

5 females and 5 males

Control animals:

not required

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: weighing on day 1 (prior to application) and on days 8 and 15, clinical examination was made several times on day of dosing (at least once during the first 30mins and with special attention given during the first 4hrs post-dose).
- Necropsy of survivors performed: yes, all gross pathological changes were recorded.
- Other examinations performed: clinical signs, body weight

Results and discussion

Effect levels

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: The test item showed no sign of acute dermal toxicity.

Mortality:

No mortalities.

Clinical signs:

other: No findings during the entire observation period.

Gross pathology:

No specific gross pathological changes were recorded for any animal, with the exception of acute injection of blood vessels in the abdominal region, which is due to the euthanasia injection.

Other findings:

Signs of irritation
- Erythema grade 1 was observed in 4/5 female and 5/5 male animals
- Erythema grade 2 was observed in 3/5 male animals
- Desquamation and eschar were observed in 5/5 female and 5/5 male animals
- Scratches were observed in 3/5 female and 5/5 male animals
- Not all signs of irritation were reversible with the observation period

Any other information on results incl. tables

TABLE: Skin Irritation – Individual Data – Females

Day after Start of Application	Animal No. 11		Animal No. 12		Animal No. 13		Animal No. 14		Animal No. 15
	E/O	Comments	E/O	Comments	E/O	Comments	E/O	Comments	E/O	Comments
day 2	1/0	nf	1/0	nf	0/0	nf	0/0	*	0/0	*
day 3	1/0	nf	1/0	nf	0/0	nf	0/0	*	0/0	*
day 4	1/0	slight s	1/0	nf	0/0	nf	0/0	*	0/0	*
day 5	0/0	s, d	1/0	d	1/0	d	0/0	slight d	0/0	d
day 6	0/0	d, es	1/0	d	1/0	d, es	0/0	slight d	1/0	d, es
day 7	0/0	d, es	0/0	d, es	0/0	d, es	0/0	d, slight s	0/0	d, s
day 8	0/0	d, es	0/0	d, es	0/0	d, es	0/0	d	0/0	slight d
day 9	0/0	slight d, slight es	0/0	d, es	0/0	d, es	0/0	d	0/0	slight d
day 10	0/0	nf	0/0	d, es	0/0	d, es	0/0	moderate d	0/0	slight d
day 11	0/0	slight d, slight s	0/0	moderate d	0/0	slight d	0/0	slight d	0/0	nf
day 12	0/0	slight d	0/0	slight d	0/0	nf	0/0	nf	0/0	nf
day 13	0/0	slight d	0/0	slight d	0/0	nf	0/0	nf	0/0	nf
day 14	0/0	slight d	0/0	nf	0/0	nf	0/0	nf	0/0	nf
day 15	0/0	nf	0/0	nf	0/0	nf	0/0	nf	0/0	nf
Comments: E = erythema; O = oedema; 1, 2, 3, 4 = scoring system laid down in OECD Guideline 404 (Table 2) d = desquamation; es = eschar; s = scratches; nf = no findings * orange discoloration at the application site, caused by the test item

 

TABLE: Skin irritation – Individual Data – Males

Day after Start of Application	Animal No. 21		Animal No. 22		Animal No. 23		Animal No. 24		Animal No. 25
	E/O	Comments	E/O	Comments	E/O	Comments	E/O	Comments	E/O	Comments
day 2	0/0	*	0/0	*	0/0	*	0/0	*	0/0	*
day 3	0/0	*	0/0	*	0/0	*	0/0	*	0/0	*
day 4	0/0	*	0/0	*	0/0	*	0/0	*	0/0	*
day 5	2/0	d, es	1/0	d, es	1/0	d, es	2/0	d, es	2/0	d, es
day 6	1/0	d, es	1/0	d, es	0/0	d, es	1/0	d, es	1/0	d, es
day 7	0/0	d, es	0/0	d, es	0/0	d, es, **	0/0	d, es, **	0/0	d, es,*
day 8	0/0	slight d, es	0/0	slight d, es	0/0	slight d, es, **	0/0	slight d, es, **	0/0	slight d, es, **
day 9	0/0	slight d, es	0/0	slight d, es, **	0/0	slight d, es, **	0/0	slight d, es, **	0/0	slight d, es, **
day 10	0/0	slight d, es	0/0	slight d, es, s	0/0	slight d, es, **	0/0	slight d, es, **	0/0	slight d, es, **
day 11	0/0	moderate d, s#	0/0	slight d	0/0	slight d, es, **, s	0/0	slight d, es, s, **	0/0	slight d, es, **, s
day 12	0/0	slight d, es, s#	0/0	slight d	0/0	slight d, es, **, s	0/0	slight es, s, **	0/0	d, es, **, s
day 13	0/0	slight d, es, s#	0/0	slight d	0/0	slight d, es, **, s	0/0	slight es, s, **	0/0	d, es
day 14	0/0	es	0/0	slight d	0/0	es	0/0	es	0/0	d, es
day 15	0/0	es	0/0	nf	0/0	es	0/0	es	0/0	nf
Comments: E = erythema; O = oedema; 1, 2, 3, 4 = scoring system laid down in OECD Guideline 404 (Table 2) d = desquamation; es = eschar; s = scratches; s#: punctiform scratches, nf = no findings * orange discoloration at the application site, caused by the test item ** small wound

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of the present study, single dermal application of phenothiazine to rats at a dose of 2000mg/kg body weight was associated with no signs of toxicity or mortality, but with signs of irritation. The dermal LD50 was determined to be greater than 2000mg/kg body weight. Phenothiazine does not require classification for acute percutaneous toxicity according to the provisions laid down in Regulation (EC) No 1272/2008.

Executive summary:

- Phenothiazine was tested for acute dermal Toxicity according to OECD guideline 402 and according to Regulation (EC) No 440/2008, Method B.3.

- Study was conducted as limit test (2000mg/kg body weight)

- 5 male and 5 female Wistar rats (Crl: WI(HAN)) were used in this study

- Corn oil was used as vehicle

- Rats were dermal exposed to phenothiazine for 24hrs

- Signs of toxicity related to dose level used, time of onset and duration were not observed (no treatment-related effects)

- No effects on organs (related to dose level) were observed

- LD50 is greater than 2000mg/kg body weight

- Signs of irritation were found in some animals. Not all signs were reversible within the observation period.

- Phenothiazine does not require classification for acute percutaneous toxicity according to the provisions laid down in Regulation (EC) No 1272/2008.