Oleic acid, compound with (Z)-N-octadec-9-enylpropane-1,3-diamine (2:1)

Endpoint summary

Description of key information

There are two acute oral studies available indicating an oral LD50 > 2000 mg/kg bw, and one acute dermal study resulting to an LD50 > 2000 mg/kg bw

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

9 Oct 2003 - 10 Dec 2003

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Well reported and carried out study according to guidelines/standards of 1996 instead of 2001 using class 200 mg/kg rather than 300 mg/kg.

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Deviations:

yes

Remarks:

version of 1996 rather than 2001 used

GLP compliance:

yes

Test type:

acute toxic class method

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
rat, Sprague-Dawley Rj: SD (IOPS Han).
- Source: Janvier, Le Genest-Saint-Isle, France.
- Age at study initiation: approximately 8 weeks old
- Weight at study initiation: 281 ± 7 g for the males and 215 ± 8 g for the females
- Fasting period before study: approx. 18 hours until 4 hours after administration
- Housing: polycarbonate cages with stainless steel lid (48 cm x 27 cm x 20 cm). Each cage contained one to seven animals of the same sex during the acclimation period and three rats of the same sex and group during the treatment period. Each cage contained autoclaved sawdust (SICSA, Alfortville, France).
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2°C
- Humidity (%): 30 to 70%
- Air changes (per hr): 12 cycles/hour of filtered, non-recycled air.
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 9 Oct 2003 To: 10 Dec 2003

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 20 mg/mL (200 mg/kg bw) and 200 mg/mL (2000 mg/kg bw)
- Amount of vehicle (if gavage): 10 mL/kg

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: No information on the toxic potential of the test item was available, for animal welfare reasons, the starting dose-level of 200 mg/kg was chosen.

Doses:

200 and 2000 mg/kg bw

No. of animals per sex per dose:

3

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations: frequently during the hours following administration and thereafter at least once per day
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight on day 1, 8 and 15 , macroscopic necropsy examination

Statistics:

Not applicable in ATC

Sex:

male/female

Dose descriptor:

LD50

Effect level:

2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: LD50 cut-off level according to OECD 423

Mortality:

At the 200 mg/kg dose-level, no mortality occurred.
At the 2000 mg/kg dose-level, no mortality occurred in males. In females, 1/3 animals died on day 2 and another one was found dead on day 8

Clinical signs:

other: At the 200 mg/kg dose-level piloerection and dyspnea were observed in all females on day 1, whereas no clinical signs were noted in males. At the 2000 mg/kg dose-level in males hypoactivity and dyspnea were recorded in all males on day 1. Then dyspnea, pi

Gross pathology:

Macroscopic examination of the main organs of the animals revealed no apparent abnormalities.

The report concluded to: Under our experimental conditions, the oral LD50 of the test item is:

- higher than 2000 mg/kg in male rats,

- comprised between 200 and 2000 mg/kg in female rats.

Interpretation of results:

Category 4 based on GHS criteria

Remarks:

Migrated information

Conclusions:

An acute oral toxicity study according to OECD 423 resulted to a LD50 between 200 -2000 mg/kgbw with LD50 cut-off value of 2000 mg/kgbw.

Executive summary:

In an OECD 423 study under GLP, Oleyl-diamine dioleate (Inipol 002) was administered to groups of 3 male and 3 female rats at dose levels of 200 and 2000 mg/kgbw formulated into corn oil applied at 10 ml/kg bw. At the 200 mg/kg dose-level, no mortality occurred. Piloerection and dyspnea were observed in all females on day 1, whereas no clinical signs were noted in males.

At the 2000 mg/kg dose-level, no mortality occurred in males. Hypoactivity and dyspnea were recorded in all males on day 1. Then dyspnea, piloerection and hypoactivity were observed in 1/3 males up to day 3, 4 or 5, respectively. In females, 1 animal died on day 2 and another one was found dead on day 8; hypoactivity, piloerection and dyspnea, together with rhinorrea and ocular secretion in the last one, were noted prior to death. In the surviving female, hypoactivity, piloerection and dypnea, together with loud breathing on days 1 and 3 and rhinorrea on days 5 and 6, were recorded from day 1 up to day 8.

At necropsy, no apparent abnormalities were observed.

The report concluded that the LD50 was higher than 2000 mg/kg in male rats, and between 200 and 2000 mg/kg in female rats. OECD 423 evaluation criteria indicate LD50 between 200 -2000 mg/kgbw with LD50 cut-off value of 2000 mg/kgbw.

Reference 2

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

5 Sep 1984 - 19 Sep 1984

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Older OECD 401 limit test, sufficiently reported.

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

no

GLP compliance:

no

Remarks:

Start GLP: Internally audited, with Study Director Statement

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Hacking and Churchill Limited, Huntingdon, Cambridgeshire, England.
- Age at study initiation: 4-6 weeks
- Weight at study initiation: ange of 102 to 132 g prior to dosing
- Fasting period before study: 4 hours (and 4 hours after dosing)
- Housing: They were housed in groups by sex in metal cages with wire mesh floors.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: minimum 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): between 21°C and 24°C
- Humidity (%): mean daily relative humidity value was 65%.
- Air changes (per hr): approximately 15 air changes/hour.
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 5 Sep 1984 To: 19 Sep 1984

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

EN 455 was administered as supplied by the Sponsor at a volume of 5.22 ml/kg (S.G. 0.957).
The appropriate dose volume of the test substance was administered to each rat using a syringe and plastic catheter.

Doses:

Pre-study: 1 and 5 g/kg bw in 2 male + 2 female per dose level.
Main study: 5 g/kg bw in 5 male and 5 female

No. of animals per sex per dose:

5 males and 5 females

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations: frequent intervals day 1 after dosing; further at least twice per day.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight on day 1, 8 and 15

Statistics:

Single dose level so no statics applied

Preliminary study:

No mortality: acute lethal oral dose was greater than 5.0 g/kg bodyweight

Sex:

male/female

Dose descriptor:

LD0

Effect level:

5 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: No mortality observed

Mortality:

No mortality

Clinical signs:

other: Pilo-erection, abnormal body carriage (hunched posture), abnormal gait (waddling), lethargy and pallor of the extremities were observed following dosing in all rats, with recovery as judged by external appearance and behaviour apparently complete on Day 4

Gross pathology:

Nothing reported

Other findings:

No

Signs of reaction to treatment observed in rats dosed orally with EN 455.

 

Table indicating observed signs in number of animals:

Signs	male	female
¿Pilo-erection	5	5
Abnormal body carriage (hunched posture)	5	5
Abnormal gait (waddling)	5	5
Lethargy	5	5
Pallor of extremities	5	5

Body weights main study:

	Bodyweight (g) at Day
Sex	1	8	15
male	124	170	243
	124	170	250
	132	166	250
	132	180	263
	124	174	257
female	104	172	207
	102	164	200
	108	124	186
	112	186	219
	108	170	207

Interpretation of results:

practically nontoxic

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Dosing of in total 7 males and 7 females of 5 g/kg body weight resulted to no mortality. The acute lethal oral dose to rats was therefore found to be greater than 5.0 g/kg bodyweight.

Executive summary:

The acute oral toxicity of EN 455 (Oleyl diamine, dioleate, purity 95%) was tested according to OECD 401 in a limit test at 5.0 g/kg bw following a preliminary test on 2 males and 2 females showing no mortality at this level.

 

The result of the preliminary study on 2 males and 2 females at 1.0 and 5.0 g/kg bw, indicated that the acute lethal oral dose was greater than 5.0 g/kg bodyweight.

Dosing was then extended to a larger group of rats (five males and five females) in order to confirm this finding.

There were no mortalities.

Pilo-erection, abnormal body carriage (hunched posture), abnormal gait (waddling), lethargy and pallor of the extremities were observed following dosing in all rats, with recovery as judged by external appearance and behaviour apparently complete on Day 4.

A poor bodyweight gain (16 g) was recorded for one female rat on Day 8. Normal bodyweight gains were recorded for all remaining rats on Days 8 and 15 (Table 3).

Conclusion: The acute lethal oral dose to rats of EN 455 was found to be greater than 5.0 g/kg bodyweight

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

Two generally straight forward studies evaluating acute toxicity via oral route performed under GLP both indicating a low level of acute toxicity.

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

01 Nov 2011 to 02 Mar 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study was performed according to current OECD/EC guidelines and according to GLP principles.

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

yes

Remarks:

study was conducted according to the acute toxic class method designed for assessment of acute oral toxicity (OECD No. 423): 2x3 females were used instead of 5 animals.

Qualifier:

according to guideline

Guideline:

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

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nohsan, Notification No. 8147, April 2011; including the most recent partial revisions.

Principles of method if other than guideline:

For treated animals, an external macroscopic examination was conducted next to an internal macroscopic examination.
The study integrity was not adversely affected by the deviation.

GLP compliance:

yes (incl. QA statement)

Remarks:

wih exception of the additional histopathology of the skin.

Test type:

other: acute toxic class method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany.
- Age at study initiation: approx. 12 weeks old
- Weight at study initiation: Body weight variation did not exceed +/- 20% of the sex mean.
- Housing: Individually in Makrolon cages
- Diet: ad libitum, pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).
- Water: ad libitum, tap water
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.3-21.9
- Humidity (%): 40-59
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: approximately 25 cm² for males and 18 cm² for females
- % coverage: appr. 10% of the total body surface
- Type of wrap if used: a surgical gauze patch (Surgy 1D), successively covered with aluminum foil and Coban elastic bandage

REMOVAL OF TEST SUBSTANCE
- Washing: with tap water
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount applied: 2.26 mL/kg bw (= 2000 mg/kg bw)

Duration of exposure:

24 hours

Doses:

2000 mg/kg

No. of animals per sex per dose:

6 females (2x3 f were dosed in a stepwise manner)

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations: mortality was checked twice daily, clinical signs once daily.
- Frequency of weighing: day 1 (pre-administration), 8 and 15.
- Necropsy of survivors performed: yes
- Other examinations performed: histopathological examination of treated skin with macroscopic abnormalities.

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred.

Clinical signs:

other: Hunched posture, lethargy, chromodacryorrhoea, piloerection and/or ptosis were shown by all animals between Days 1 and 10. The treated skin-area of all animals showed general or focal erythema, scales, scabs, a thickened area and/or fissures during the ob

Gross pathology:

Three animals for which an external macroscopic examination was conducted showed scab formation on the dorso-lumbar region of the skin. No other abnormalities were found at macroscopic post mortem examination of the animals.

Other findings:

Treatment-related microscopic skin findings were as follows:
- Ulceration (characterized by full thickness loss of the epithelium through the basement membrane) was present in 5/6 2000 mg/kg treated rats at slight (3/6) or marked (2/6) degree.
- A fibrinous exudate (a focally fibrillar eosinophilic mass of exudative proteinaceous material and cellular debris) was present on the skin surface of all rats at a minimal (1/6), slight (1/6) or moderate (4/6 degree). The most severe exudate was seen in ulcerated areas.
- Acanthosis, thickening/hyperplasia of the epidermis (increase in the number of epithelial cell layers which may extend into the dermis), was present in the non-ulcerated /less severely affected areas of all rats at a slight (5/6) or moderate (1/6) degree. This was likely a reactive response to the test item.
- All rats had lymphogranulocytic inflammation (characterized by a mixture of predominately lymphocytic and some granulocytic inflammatory cells) present in the dermis at minimal (3/6), slight (1/6) or moderate (2/6) degree. Inflammation extended focally into the underlying panniculus muscle which separates the dermis from the underlying subcutaneous tissue.

Interpretation of results:

not classified

Remarks:

Migrated information for dermal acute toxicity Criteria used for interpretation of results: EU

Conclusions:

The dermal toxicity of oleyl diamine was tested according to current OECD/EC guidelines and under GLP principles. The LD50 was determined to be > 2000mg/kg.

Based on these results, Oleyl diamine, dioleate does not have to be classified and has no obligatory labelling requirement for acute dermal toxicity according to the:
- Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2007),
- Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures.

Executive summary:

Assessment of acute dermal toxicity with Oleyl diamine, dioleate in the rat.

The study was carried out based on the guidelines described in:

-      OECD No.423 (2001) "Acute Oral Toxicity, Acute Toxic Class Method"

-      Commission Regulation (EC) No 440/2008, B1tris: "Acute Oral Toxicity, Acute Toxic Class Method"

-      EPA, OPPTS 870.1100 (2002), "Acute Oral Toxicity"

-      JMAFF guidelines (2011) including the most recent partial revisions.

-      OECD No.402 (1987) "Acute Dermal Toxicity"

-      Commission Regulation (EC) No 440/2008, B3: "Acute Toxicity (Dermal)"

-      EPA, OPPTS 870.1200 (1998), "Acute Dermal Toxicity"

Oleyl diamine, dioleate was administered to two subsequent groups of three female Wistar rats by a single dermal application at 2000 mg/kg body weight for 24 hours. Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice (Day 15). Histopathology was conducted on the treated skin area from all rats.

Results: Hunched posture, lethargy, chromodacryorrhoea, piloerection and/or ptosis were shown by all animals between Days 1 and 10. The treated skin-area of all animals showed general or focal erythema, scales, scabs, a thickened area and/or fissures during the observation period.

The changes noted in body weight gain in males and females were within the range expected for rats used in this type of study and were therefore considered not indicative of toxicity.

Three animals for which an external macroscopic examination was conducted showed scab formation on the dorso-lumbar region of the skin. No other abnormalities were found at macroscopic post mortem examination of the animals.

Treatment-related histopathological findings in the treated skin area consisted of ulceration (complete loss of the epidermis) in most rats with a superficial fibrinous exudate (this correlated with the scab formation seen macroscopically) in all rats. There was hyperplasia of the epidermis (acanthosis) in the non-ulcerated areas of all rats. Furthermore, dermal lymphogranulocytic inflammation was present in all rats.

 

The dermal LD50 value of Oleyl diamine, dioleate in Wistar rats was established to exceed 2000 mg/kg body weight.

 

From the histopathological results it is concluded that Oleyl diamine, dioleate is an irritant and has corrosive potential when applied with an occlusive dressing to the rat skin at 2000 mg/kg for 24 hours.

Based on these results, Oleyl diamine, dioleate does not have to be classified and has no obligatory labeling requirement for acute dermal toxicity according to the:

- Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2007),

- Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

Recent, high quality study.

Additional information

Acute oral toxicity:

In an OECD 423 study under GLP, Oleyl-diamine dioleate was administered to groups of 3 male and 3 female rats at dose levels of 200 and 2000 mg/kgbw, formulated into corn oil applied at 10 mL/kg bw. At the 200 mg/kg dose-level, no mortality occurred. Piloerection and dyspnoea were observed in all females on day 1, whereas no clinical signs were noted in males.

At the 2000 mg/kg dose-level, no mortality occurred in males. Hypoactivity and dyspnoea were recorded in all males on day 1. Then dyspnoea, piloerection and hypoactivity were observed in 1/3 males up to day 3, 4 or 5, respectively. In females, 1 animal died on day 2 and another one was found dead on day 8; hypoactivity, piloerection and dyspnoea, together with rhinorrea and ocular secretion in the last one, were noted prior to death. In the surviving female, hypoactivity, piloerection and dyspnoea, together with loud breathing on days 1 and 3 and rhinorrea on days 5 and 6, were recorded from day 1 up to day 8.

At necropsy, no apparent abnormalities were observed.

The report concluded that the LD50 was higher than 2000 mg/kg in male rats, and between 200 and 2000 mg/kg in female rats. OECD 423 evaluation criteria indicate LD50 between 200 -2000 mg/kgbw with LD50 cut-off value of 2000 mg/kgbw.

 

In another study according to OECD 401 and under GLP, Oleyl-diamine dioleate (95% purity) was administered undiluted at 5000 mg/kg bw (5.22 mL/kg) to a group of 5 male and 5 female rats following a trial test in 2 male and 2 female rats at 1000 and 5000 mg/kg bw showing no mortality. Also the main study resulted to no mortality. All animals showed signs of pilo-erection, hunched posture, abnormal gait (waddling), lethargy and pallor of extremities.

 

The differences in results between the two studies might possibly be related to the differences in dosing scheme applied. In the first study animals were dosed in a formulation of about 20% in corn oil, whereas in the second study the substance was administered pure. As Oleyl-diamine dioleate is not soluble in water, but soluble in oil, the solution in oil possibly could lead to an enhanced bioavailability and faster absorption kinetics.

 

The first study resulted to an overall mortality of 2/6 animals, both females. Although this indicates an overall LD50 > 2000 mg/kg, the formal and conservative evaluation of this study design using only a very small number of animals and thus high level of uncertainty, and no availability of possible sex-differences, would lead to classification Cat.4 with LD50 between 200(300) - 2000 mg/kg bw, with aLD50 cut-off level of 2000 mg/kg based on the deaths seen in the females.

However, there is an additional study of similar validity but applying larger number of animals at higher dose levels, showing an even higher LD50 in excess of 5000 mg/kg.

This second study, and additionally a 14-day range finding and a 28-day study, all show that there is no principal difference in susceptibility between male and female animals after all, and thus even the first study can be reported to show that the LD50 > 2000 mg/kg. Hence no classification for acute oral toxicity is needed.

 

Acute dermal toxicity:

Acute dermal toxicity of Oleyl-diamine dioleate was evaluated following a single dermal application at 2000 mg/kg body weight for 24 hours in two subsequent groups of three female Wistar rats. Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice (Day 15). Histopathology was conducted on the treated skin area from all rats.

All animals showed hunched posture, lethargy, chromodacryorrhoea, piloerection and/or ptosis between Days 1 and 10. The treated skin-area of all animals showed general or focal erythema, scales, scabs, a thickened area and/or fissures during the observation period.

Treatment-related histopathological findings in the treated skin area consisted of ulceration (complete loss of the epidermis) in most rats with superficial fibrinous exudates, correlating with the scab formation seen macroscopically, in all rats.

The dermal LD50 value of Oleyl-diamine dioleate in Wistar rats was established to exceed 2000 mg/kg body weight.

 

Acute inhalation toxicity:

No data available. Due to irritating properties respiratory irritation would be in principle possible. Due to the low vapour pressure (< 0.0015 Pa at 20°C based on C12/14 -diamine rather than Oleyl diamine dioleate salt) inhalation of vapours leading to irritation of airways will not occur. The indicated use pattern also does not give rise to a high concern for specific exposures via inhalation of these oleyl-diamine dioleate salts.

Justification for selection of acute toxicity – oral endpoint
Study showing the most severe results from the two available studies of comparable validity.

Justification for selection of acute toxicity – inhalation endpoint
No inhalation studies are needed when vp < 0.1 Pa at 20°C

Justification for selection of acute toxicity – dermal endpoint
Only study available.

Justification for classification or non-classification

The evaluation of available data leads to the conclusion that for oleyl-diamine dioleate, both the acute oral and acute dermal LD50 are determined to be above 2000 mg/kg bw. Consequently, oleyl-diamine dioleate does not need to be classified for acute oral or dermal toxicity for EU-CLP.

For inhalation route no data is available. However, in view of low likelihood of exposures related to very low vapour pressure no further testing is needed.

Available information indicates that oleyl-diamine dioleate does not require classification for aspiration hazard.