N-Phenyl-diethanolamine, reaction products with formaldehyde

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

14 August 2020 - 09 September 2020

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Test conducted in accordance with OECD Test Guideline and in accordance with GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2020

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity: Fixed Dose Procedure)

Version / remarks:

09 October 2017

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

yes

Test material

Test material information

Constituent 1

Test material form:

liquid

Details on test material:

Storage conditions: At room temperature protected from light
Physical description: Light orange liquid

Test animals

Species:

rat

Strain:

Wistar

Remarks:

Crl: WI(Han)

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Females nulliparous and non-pregnant: yes
- Rationale for use of males (if applicable): N/A
- Age at study initiation: Single young adult animal (approx. 10 weeks old)
- Weight at study initiation: 181 g
- Fasting period before study: Not specified
- Housing: On arrival, the animal was group housed (up to 5 animals of the same sex together) in polycarbonate cages (Makrolon MIV type; height 18 cm.) and following assignment to the study, the animal was individually housed in polycarbonate cages (Makrolon MIII type; height 18 cm.) containing sterilized wooden fibers as bedding material (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany) equipped with water bottles. The room in which the animal was kept was documented in the study records. The cage was clearly labeled.
- Historical data:
- Diet: Pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany) was provided ad libitum throughout the study, except during designated procedures.
The feed was analyzed by the supplier for nutritional components and environmental contaminants. Results of the analysis were provided by the supplier and are on file at the Test Facility. It is considered that there were no known contaminants in the feed that would interfere with the objectives of the study.
- Water (e.g. ad libitum): Municipal tap-water was freely available to each animal via water bottles. Periodic analysis of the water was performed, and results of these analyses are on file at the Test Facility. It is considered that there were no known contaminants in the water that would interfere with the objectives of the study.
- Acclimation period: The animal was allowed to acclimate to the Test Facility toxicology accommodation for at least 5 days before the commencement of dosing.
- Microbiological status when known: Not reported
- Method of randomisation in assigning animals to test and control groups:
Animals were assigned to the study at the discretion of the coordinating biotechnician, with all animals within ± 20% of the sex mean body weights. Animals in poor health or at extremes of body weight range were not assigned to the study.
Before the initiation of dosing, a health inspection was performed, and any assigned animal considered unsuitable for use in the study were replaced by alternate animals obtained from the same shipment and maintained under the same environmental conditions.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Actual daily mean temperature during study period: 21-22°C
- Humidity (%): Actual daily mean relative humidity during study period: 48 -70%
- Air changes (per hr): => 10/h with 100% fresh air (no air recirculation)
- Photoperiod (hrs dark / hrs light): 12 hour light/12 hour dark cycle
- Animal enrichment: For psychological/environmental enrichment, the animal was provided with paper (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom)

IN-LIFE DATES: From: 14 August 2020 To: 09 September 2020

Administration / exposure

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: The test item was applied to approx. 18 cm² .
- % coverage: Approx. 10 %
- Type of wrap if used: The test item was held in contact with the skin with a dressing, consisting of a surgical gauze patch (Surgy 1D), successively covered with Coban elastic bandage. A piece of Micropore tape was additionally used for fixation of the bandages.

REMOVAL OF TEST SUBSTANCE
- Washing (if done):
- Time after start of exposure:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit):
- Concentration (if solution):
- Constant volume or concentration used: yes/no
- For solids, paste formed: yes/no

VEHICLE
- Amount(s) applied (volume or weight with unit):
- Concentration (if solution):
- Lot/batch no. (if required):
- Purity:

Duration of exposure:

Application period: 24 h, after which the dressing was removed, and the skin cleaned of residual test item using water.

Doses:

2000 mg/kg

The dose volume for the animal was based on the body weight measurement prior to dosing. Dose volume (mL/kg body weight) was calculated as follows: Dose level (g/kg) / spec.gravity or density (g/mL) * purity correction factor.

Animal welfare reasons prevented the initiation of the main study due to severe skin effects, therefore no addiitonal doses were applied.

No. of animals per sex per dose:

A single female was dosed at the single 2000 mg/kg bw dose.

Control animals:

other: Adjacent areas of untreated skin of the animal served as a control

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Throughout the study, the animal was observed for general health/mortality and moribundity twice daily, in the morning and at the end of the working day. Post-dose observations were performed at periodic intervals on the day of dosing (at least three times) and once daily thereafter. The animal was examined for reaction to dosing. The onset, intensity and duration of these signs was recorded (if appropriate), particular attention being paid to the animal during and for the first hour after dosing.
The animal was weighed individually on Day 1 (pre-dose), 8 and 15.
- Necropsy of survivors performed: yes
- Clinical signs including body weight: yes
- Other examinations performed: clinical signs, macroscopic findings

Statistics:

Not applicable, only a single dose was applied to a single animal during the test

Results and discussion

Preliminary study:

The results from the single animal dosed at 2000 mg/kg are as follows:
No mortality occurred.
Systemic signs of toxicity consisted of hunched posture on Days 3-5.
Local toxicity consisted of very slight to well-defined erythema between 24- and 72-hours post-dosing. Red discoloration was seen between Days 2 and 7, which resulted in flaking, signs of necrosis and/or scabs on the lumbar skin / right flank between Days 2 and 15.
The body weight gain shown by the animal during the observation period was within the range expected for rats used in this type of study.
No abnormalities were found at macroscopic post-mortem examination of the animal.

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

no indication of skin irritation up to the relevant limit dose level

Remarks:

Minor transient systemic toxicity (hunched posture) was observed and mortality is not to be expected if additional animals were dosed.

Mortality:

None

Clinical signs:

Minor transient systemic toxicity (hunched posture) was observed on Days 3-5.
Local toxicity consisted of very slight to well-defined erythema between 24- and 72-hours post-dosing. Red discoloration was seen between Days 2 and 7, which resulted in flaking, signs of necrosis and/or scabs on the lumbar skin / right flank between Days 2 and 15.

Body weight:

The body weight gain shown by the animal during the observation period was within the range expected for rats used in this type of study.

Gross pathology:

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

Other findings:

N/A

Any other information on results incl. tables

Table 1. Individual Mortality - Female Day(s): - Relative to Start Date

2000 mg/kg Group 1
	Day of Death	Removal Date	Path Removal Reason
1	15	09-Sep-2020	Terminal euthanasia

 

Table 2. Individual Clinical Observations

Group 1 Sex: Female 2000 mg/kg
	Observation Type: All Types	Day(s) Relative to Start Date
		1 0h	1 2h	1 4h	2	3	4	5	6	7	8	9	10	11	12	13	14	15
1	Hunched Posture	.	.	.	.	X	X	X	.	.	.	.	.	.	.	.	.	.
	Skin, Flaking, Lumbar, Grade 1	.	.	.	.	.	X	X	X	.	.	.	.	.	X	.	X	X
	Skin, Flaking, Lumbar, Grade 2	.	.	.	.	.	.	.	.	X	X	X	X	.	.	X	.	.
	Skin, Flaking, Lumbar, Moderate	.	.	.	.	.	.	.	.	.	.	.	.	X	.	.	.	.
	Skin, Discolored, Lumbar, Red	.	.	.	X	X	X	X	X	X	.	.	.	.	.	.	.	.
	Skin, Scab, Lumbar	.	.	.	.	X	X	X	X	X	X	X	X	X	X	X	X	X

X = present

Table 3. Individual Body Weights - Female

2000 mg/kg Group 1	Day(s) Relative to Start Date
	1	8	15
1	181	193	203
Mean	181.0	193.0	203.0
SD	-	-	-
N	1	1	1

 

Table 4. Irritation Observations

Group 1 Sex: Female 2000 mg/kg
	Observation Type: All Types	Day(s) Relative to Start Date
		3 24H	4 48H	5 72H
1	Erythema, Lumbar, Grade 1	X	X	.
	Erythema, Lumbar, Grade 2	.	.	X
	Other (see comment), Right Subcapsular^a	X	X	X

^a Scab on right flank showing necrosis.

X = Present

Individual Macroscopic Pathology:

Animal: 1, Group: 1, Sex: Female, Dose: 2000 mg/kg, Removal Reason: Terminal Euthanasia

Study Day (Week) of Death: 15 (3)

Gross Status: Complete

Gross Pathology Observations [Correlation]:

No observations found

Gross Pathology - The following Tissues were Not Examined:

None

 

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The dermal LD50 value of the test item in Wistar Han rats was established to be > 2000 mg/kg body weight.

Executive summary:

A test was performed in accordance with OECD 402 (2017), in order to determine the potential toxicity of the test item, when given by a single dermal dose.

Initially, the test item was administered to a single female Wistar Han rat by a single dermal application at 2000 mg/kg body weight for 24 hours in a range finder study. The animal was subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice (Day 15).

No mortality occurred. Furthermore, the body weight gain shown by the animal during the observation period was within the range expected for rats used in this type of study and no abnormalities were found at macroscopic post-mortem examination. Hunched posture was noted between Days 3 and 5. Local toxicity consisted of very slight to well-defined erythema between 24- and 72-hours post-dosing. Red discoloration was seen between Days 2 and 7, which resulted in flaking, signs of necrosis and/or scabs on the lumbar skin / right flank between Days 2 and 15. Based on the severity of these effects, it was considered that the other two animals assigned to the main study could not be dosed for animal welfare reasons.

Based on the results, it can be stated that the dermal LD50 value exceeds 2000 mg/kg, because only minor transient systemic toxicity (hunched posture) was observed and mortality is not to be expected if additional animals were dosed.

Based on these results, the test item would not be classified in accordance with Regulation (EC) No 1272/2008 on Classification, Labelling and Packaging of Substances and Mixtures.