2,2'-(octylimino)bisethanol

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2015-12-07 to 2016-04-13

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

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:

(Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, München, Germany)

Test type:

standard acute method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

2,2'-(octylimino)bisethanol

EC Number:

239-555-0

EC Name:

2,2'-(octylimino)bisethanol

Cas Number:

15520-05-5

Molecular formula:

C12H27NO2

IUPAC Name:

2-[(2-hydroxyethyl)(octyl)amino]ethan-1-ol

Test material form:

other: liquid

Details on test material:

Name: Genamin 3920
CAS No.: 15520-05-5
Molecular Weight of Base Form: 217.3483
Chemical Name: 2,2’-(Octylimino)bisethanol
Physical State at RT: liquid
Colour: clear
pH: 11.6 (23 °C, water:ethanol 1:1)

Storage Conditions: 2 – 8 °C, protected from light

Safety Precautions: The routine hygienic procedures are sufficient to assure personnel health and safety.

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

Housing and Feeding Conditions
- Full barrier in an air-conditioned room
- Temperature: 22 +/- 3 °C
- Relative humidity: 55 +/- 10%
- Artificial light, sequence being 12 hours light, 12 hours dark
- Air change: 10 x / hour
- Free access to Altromin 1324 maintenance diet for rats and mice (lot no. 02102150820)
- Free access to tap water, sulphur acidified to a pH value of approximately 2.8 (drinking water, municipal residue control, microbiological controls at regular intervals)
- The animals were kept individually in IVC cages, type III H, polysulphone cages on Altromin saw fibre bedding (lot no. 0446 )
- Certificates of food, water and bedding are filed for two years at BSL Munich and afterwards archived at Eurofins Munich
- Adequate acclimatisation period (at least five days) under laboratory conditions

Administration / exposure

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

Preparation of the Animals:
The animals were marked for individual identification by tail painting.
Approximately 25 hours before the test, the fur was removed from the dorsal area of the trunk using an electric clipper. Care was taken to avoid
abrading the skin, and only animals with healthy intact skin were used.
No less than 10% of the body surface was cleared for the application.
Prior to the application a detailed clinical observation was made of all animals. Only healthy animals were used.

Application:
The test item was applied at a single dose, uniformly over an area which was approximately 10% of the total body surface.
The test item was held in contact with the skin by a dressing throughout a 24-hour period. The dressing consisted of a gauze-dressing and
non-irritating tape and was fixed with an additional dressing in a suitable manner.

Duration of exposure:

The test item was held in contact with the skin throughout a 24-hour period. At the end of the exposure period the residual test item was not
removed.

Doses:

The test item was applied at a single dose of 2000 mg/kg body weight to each animal.

No. of animals per sex per dose:

5 male and 5 female

Control animals:

not required

Details on study design:

Observation period:
All animals were observed for 14 days after dosing

Weight Assessment:
The animals were weighed on day 1 (prior to the application) and on days 8 and 15.

Clinical Examination:
careful clinical examination was made several times on the day of dosing (at least once during the first 30 minutes and with special attention given
during the first 4 hours post-dose). As soon as symptoms were noticed they were recorded. Thereafter, the animals were observed for clinical signs once daily until the end of the observation period. All abnormalities were recorded.
Cageside observations included changes in the skin and fur, eyes and mucous membranes. Also respiratory, circulatory, autonomic and central
nervous systems and somatomotor activity and behaviour pattern were examined. Attention was directed to observations of tremors, convulsions,
salivation, diarrhoea, lethargy, sleep and coma.

Pathology:
At the end of the observation period the animals were sacrificed with an overdosage of pentobarbital injected intraperitoneally (Narcoren®, Merial;
lot no.: 250055; expiry date: 31/05/2018) at a dosage of 250-400 mg/kg bw. All animals were subjected to gross necropsy and examined
macroscopically for gross pathological changes. In absence of gross pathological changes no tissues were preserved for a possible histopathologicalevaluation.

Evaluation of Results:
Individual reactions of each animal were recorded at each time of observation.
Toxic response data were recorded by sex and dose level.
Nature, severity and duration of clinical observations were described.
The body weight changes were summarised in a tabular form.
Necropsy findings were described.
With few exceptions, data were captured using the validated departmental computerised system E WorkBook (version 9.4.0, ID Business Solutions Ltd.)

Statistics:

According to OECD guidelines, the biological relevance of the results is the criterion for the interpretation of results, a statistical evaluation of the
results is not regarded as necessary.

Results and discussion

Effect levels

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality was observed

Clinical signs:

other: No treatment-related effects were observed.

Gross pathology:

On the skin and in the subcutis either scab or scab and one or more red or dark foci were observed in all animals.

Other findings:

Erythema up to grade 2 and oedema up to grade 2 were observed in all animals. Either necrosis or eschar or both were observed in all animals.
The signs of irritation were not reversible within the observation period.

Any other information on results incl. tables

Clinical Signs of Systemic Toxicity - Individual Data - Males

Animal No.		Dose (mg/kg bw)		Timepoint		Observations		Comment
21		2000		(0min – 240min)		Nsf		-
21		2000		(2d – 15d)		Nsf		Local findings
22		2000		(0min – 240min)		Nsf		-
22		2000		(2d- 5d)		Nsf		Local findings
22		2000		(6d – 12d)		Nsf		-
22		2000		(13d – 15d)		Nsf		Local findings
23		2000		(0min – 6d)		Nsf		-
23		2000		(7d)		Nsf		Local findings
23		2000		(8d – 12d)		Nsf		-
23		2000		(13d – 15d)		Nsf		Local findings
24		2000		(0min – 6d)		Nsf		-
24		2000		(7d – 15d)		Nsf		Local findings
25		2000		(0min – 240min)		Nsf		-
25		2000		(2d – 15d)		Nsf		Local findings

min = minute(s); h = hour(s); d = day(s), bw = body weight;nsf = no specific findings Clinical Signs of Systemic Toxicity - Individual Data - Females

Animal No.		Dose (mg/kg bw)		Timepoint		Observations		Comment
26		2000		(0min – 240min)		Nsf		-
26		2000		(2d – 15d)		Nsf		Local findings
27		2000		(0min- 12d)		Nsf		-
27		2000		(13d – 15d)		Nsf		Local findings
28		2000		(0min – 240min)		Nsf		-
28		2000		(2d – 15d)		Nsf		Local findings
29		2000		(0min – 240min)		Nsf		-
29		2000		(2d – 15d)		Nsf		Local findings
30		2000		(0min – 240 min)		Nsf		-
30		2000		(2d – 15d)		Nsf		Local findings

min = minute(s); h = hour(s); d = day(s), bw = body weight;nsf = no specific findings

Skin Irritation at Application Site – Individual Data – Males

Animal No.	Timepoint	Oedema/	Local	Comment
		Erythema	Skin Findings
21	(2d – 5d)	0/0	Necrosis	Black focus at application site approx. 2cmx2cm
21	(6d – 8d)	1/2	Necrosis	Black focus approx. 3cmx4cm
21	(9d -12d)	1/1	Necrosis	Black focus approx. 3cmx4cm
21	(13d – 15d)	0/1	Necrosis	Eschar peeling off, black focus approx. ø 1cm
22	(2d)	0/0	Necrosis	Black focus at application site approx. 2cmx2cm
22	(3d – 4d)	0/0	Necrosis	Black focus at application site approx. ø 0.5cm
22	(5d)	1/1	Necrosis	Black focus at application site approx. ø 0.5cm
22	(6d)	2/2	Nsf	-
22	(7d)	1/2	Nsf	-
22	(8d – 12d)	0/1	Nsf	-
22	(13d - 14d)	0/0	Eschar	3 foci ø 0.5cm
22	(15d)	0/0	Eschar	2 foci ø 0.3cm
23	(2d – 4d)	0/0	Nsf	-
23	(5d)	1/1	Nsf	-
23	(6d)	2/2	Nsf	-
23	(7d)	2/2	Necrosis	Black focus approx. 1.5cmx1.5cm
23	(8d – 12d)	0/1	Nsf	-
23	(13d)	0/0	Eschar	1 focus ø 0.5 cm
23	(14d – 15d)	0/0	Eschar	1 focus ø 0.3 cm
24	(2d – 4d)	0/0	Nsf	-
24	(5d)	1/1	Nsf	-
24	(6d)	2/2	Nsf	-
24	(7d)	2/2	Necrosis	Black focus approx. 2cmx2cm
24	(8d)	1/1	Necrosis	Black focus approx. 1cmx1cm
24	(9d – 12d)	0/1	Necrosis	Black focus approx. 1cmx1cm
24	(13d)	0/0	Eschar	1 focus ø 0.5cm
24	(14d – 15d)	0/0	Eschar	1 focus ø 0.3cm
25	(2d)	0/0	Necrosis	Black focus at application site approx. 2cmx2cm
25	(3d – 4d)	0/0	Necrosis	Black focus at application site approx. 1cm
25	(5d)	1/1	Necrosis	Black focus at application site approx. 1cm
25	(6d)	1/2	Necrosis	Black focus approx. 1cmx0.5cm
25	(7d)	1/2	Necrosis	Black focus approx. 2cmx2cm
25	(8d)	1/1	Necrosis	Black focus approx. 1.5cmx1.5cm
25	(9d – 12d)	0/1	Necrosis	Black focus approx. 1.5cmx1.5cm
25	(13d – 15d)	0/0	Eschar	2 foci ø 0.3cm

E = erythema; O = oedema; 0, 1, 2, 3, 4 = scoring system laid down in OECD Guideline 404 Skin Irritation at Application Site - Individual Data - Females

Animal No.	Timepoint	Oedema/	Local	Comment
		Erythema	Skin Findings
26	(2d – 4d)	0/0	Necrosis	Black focus at application site approx. 2cmx2cm
26	(5d)	2/2	Necrosis	Black focus at application site approx. 2cmx2cm
26	(6d – 12d)	2/2	Necrosis	Black focus approx. 3cmx4cm
26	(13d – 15d)	0/1	Necrosis	Black focus approx. 2cmx2cm, eschar peeling off
27	(2d – 3d)	0/0	nsf	-
27	(4d)	0/2	nsf	-
27	(5d)	1/2	nsf	-
27	(6d – 7d)	2/2	nsf	-
27	(8d – 12d)	0/1	nsf	-
27	(13d – 15d)	0/0	Eschar	1 focus approx. ø 0.4cm
28	(2d – 4d)	0/0	Necrosis	Black focus at application site approx. 2cmx2cm
28	(5d)	1/2	Necrosis	Black focus at application site approx. 2cm.x2cm
28	(6d – 7d)	2/2	Necrosis	Black focus approx. 2cmx2cm
28	(8d – 12d)	1/1	Necrosis	Black focus approx. 2cmx2cm
28	(13d – 15d)	0/1	Necrosis	Black focus approx. 2cmx2cm
29	(2d – 3d)	0/0	Necrosis	Black focus at application site approx. 2cmx2cm
29	(4d)	0/1	Necrosis	Black focus at application site approx. 2cmx2cm
29	(5d)	0/2	Necrosis	Black focus at application site approx. 2cmx2cm
29	(6d)	1/2	Necrosis	Black focus approx. 2cmx2cm
29	(7d)	1/2	Necrosis	Black focus approx. 2.5cmx2.5cm
29	(8d)	1/1	Necrosis	Black focus approx. 2.5cmx2.5cm
29	(9d – 12d)	0/1	Necrosis	Black focus approx. 2.5cmx2.5cm
29	(13d)	0/0	Necrosis	Black focus approx. 2.5cmx2.5cm
29	(14d – 15d)	0/0	Eschar	Eschar peeling off, approx. ø 1cm
30	(2d – 4d)	0/0	Necrosis	Black focus at application site approx. 2cmx2cm
30	(5d – 6d)	1/2	Necrosis	Black focus at application site approx. 2cmx2cm
30	(7d)	2/2	Necrosis	Black focus approx. 2.5cmx2.5cm
30	(8d – 12d)	1/1	Necrosis	Black focus approx. 2.5cmx2.5cm
30	(13d – 14d)	0/1	Necrosis	Black focus approx. 1cmx1cm, eschar peeling off
30	(15d)	0/0	Eschar	Eschar peeling off, approx. ø 1cm

E = erythema; O = oedema; 0, 1, 2, 3, 4 = scoring system laid down in OECD Guideline 404;
nsf = no specific findings

Absolute Body Weights in g and Body Weight Gain in %

Test Group	Animal No.	Dose (mg/kg bw)	Body Weight (g) on Day			BW gain in Comparison to Day 1 (%)
			Day 1	Day 8	Day 15	Day 15
Males	21	2000	243	252	258	6
	22		241	257	278	15
	23		244	257	280	15
	24		267	283	321	20
	25		241	257	282	17
Females	26	2000	231	227	212	-8
	27		232	228	235	1
	28		225	221	227	1
	29		230	237	243	6
	30		220	218	225	2

bw = body weight

Macroscopic Findings - Individual Data - Males and Females

Test Group	Animal No.	Dose (mg/kg bw)	Organ	Macroscopic Findings
Males	21	2000	Skin	Scab 1cm
			Subcutis	Focus red 1cm
	22	2000	Skin	Scab 2cmx0.3cm
	23	2000	Skin	Scab 0.3cm
	24	2000	Skin	Scab 0.3cm
	25	2000	Skin	Scab 2cmx0.3cm
Females	26	2000	Skin	Scab 4cm
			Subcutis	Focus dark 1cm;
				focus red 3cm
	27	2000	Skin	Scab 0.4cm
			Subcutis	Focus red 0.2cm
	28	2000	Skin	Scab 4cm
			Subcutis	Focus red 1cm
	29	2000	Skin	Scab 1cm
			Subcutis	Focus red 1cm
	30	2000	Skin	Scab 1cm
			Subcutis	Focus red 1cm

nsf = no specific findings

LD₅₀

Test Group	Dose (mg/kg bw)	Number of Animals	Number of Intercurrent Deaths	LD50Cut-Off             (mg/kg bw)
Males	2000	5	0	> 2000
Females	2000	5	0

bw = body weight

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Remarks:

severely irritating but no systemic toxicity

Conclusions:

Under the conditions of the present study, single dermal application of the test item Genamin 3920 to rats at a dose of 2000 mg/kg body weight was
associated with no mortality and no signs of systemic toxicity but with signs of irritation/corrosion at the application site which were not fully
reversible within the observation period.
The dermal LD50 was determined to be > 2000 mg Genamin 3920 / kg body weight.

Executive summary:

Summary Results

LD₅₀:                                      > 2000 mg/kg bw

Species/strain:                      WISTAR Crl: WI(Han) rats

Vehicle (moistening):             no vehicle used

Number of animals:                5 male and 5 female

Duration of exposure:           24 hours

Method:                                OECD 402, EC 440/2008, Method B.3

Results

Test Group		Animal No.		Dose (mg/kg bw)		Number of Animals		Number of Intercurrent Deaths
Males		21 - 25		2000		5		0
Females		26 - 30		2000		5		0

bw = body weight

Signs of systemic toxicity related to dose level used, time of onset and duration:

No treatment-related effects were observed.

Effect on organs (related to dose level):

On the skin and in the subcutis either scab or scab and one or more red or dark foci were observed in all animals.

Signs of irritation:

Erythema up to grade 2 and oedema up to grade 2 were observed in all animals.Either necrosis or eschar or both were observed in all animals.

The signs of irritation were not reversible within the observation period.

Conclusion

Under the conditions of the present study, single dermal application of the test itemGenamin 3920to rats at a dose of 2000 mg/kg body weight was associated with no mortality and no signs of systemic toxicity but with signs of irritation/corrosion at the application site which were not fully reversible within the observation period.

The dermal LD₅₀was determined to be > 2000 mgGenamin 3920/ kg body weight.