Amidation products of C16-18 (even numbered), C18 unsaturated fatty acids esters with 1,1'-iminodipropan-2-ol

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

24 August 2016 to 20 September 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

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)

Version / remarks:

Organization for Economic Co-operation and Development (OECD), OECD Guidelines for Testing of Chemicals, Section 4, Health Effects, No. 402, "Acute Dermal Toxicity", Paris, 1987.

Deviations:

yes

Remarks:

See "Any other information" for details

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

Commission Regulation (EC) No 440/2008 Part B: Methods for the Determination of Toxicity and other Health Effects; B.3: "Acute Toxicity (Dermal)". Official Journal of the European Union No. L142, May 2008, including most recent amendments.

Deviations:

yes

Remarks:

See "Any other information" for details

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Version / remarks:

United States Environmental Protection Agency (EPA). Health Effects Test Guidelines, OPPTS 870.1200, Acute Dermal Toxicity. Office of Prevention, Pesticides and Toxic Items (7101), EPA 712-C-98-192, August 1998.

Deviations:

yes

Remarks:

See "Any other information" for details

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: JMAFF, Notification No 8147

Version / remarks:

Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nousan, Notification No 8147, November 2000; including the most recent partial revisions.

Deviations:

yes

Remarks:

See "Any other information" for details

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Amidation products of C16-18 (even numbered), C18 unsaturated fatty acids esters with 1,1'-iminodipropan-2-ol

EC Number:

701-003-6

Cas Number:

1454803-04-3

Molecular formula:

C20H39NO3 to C26H51NO3

IUPAC Name:

Amidation products of C16-18 (even numbered), C18 unsaturated fatty acids esters with 1,1'-iminodipropan-2-ol

Test material form:

liquid

Details on test material:

Identification: MLA-3202
Appearance: Clear amber-red liquid
Purity/Composition: UVCB
Test item storage: At room temperature
Stable under storage conditions until 17 February 2019 (expiry date)
Purity/composition correction factor: No correction factor required
Chemical name (IUPAC), synonym or trade name: Amides, tallow, N,N-bis(2-hydroxypropyl)
CAS Number: 1454803-04-3
Test item handling No specific handling conditions required

Specific details on test material used for the study:

Batch: RC-1045Study specific test item informationPurity/composition correction factor: No correction factor requiredChemical name (IUPAC), synonym or trade name: Amides, tallow, N,N-bis(2-hydroxypropyl)CAS Number: 1454803-04-3Test item handling: No specific handling conditions required

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

Species: Rat, Wistar strain, Crl:WI (Han) (outbred, SPF-Quality). Recognized by international guidelines as the recommended test system (e.g. OECD, EC). Source: Charles River Deutschland, Sulzfeld, Germany. Number of animals: 5 males and 8 females (females were nulliparous and non-pregnant). Age and body weight: Young adult animals (approx. 10 weeks old) were selected. Body weight variation did not exceed +/- 20% of the sex mean. Identification: Tail mark with indelible ink. Health inspection: At least prior to dosing. It was ensured that the animals were healthy and that the skin to be treated was intact and free from any abnormality.Animal Husbandry Conditions: Environmental controls for the animal room were set to maintain 18 to 24°C, a relative humidity of 40 to 70%, at least 10 air changes/hour, and a 12-hour light/12-hour dark cycle. Any variations to these conditions were maintained in the raw data and had no effect on the outcome of the study. Accommodation: Individually housed in labeled Makrolon cages (MIII type, height 18 cm.) containing sterilized sawdust as bedding material (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany) and paper as cage-enrichment (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom). Acclimatization period was at least 5 days before start of treatment under laboratory conditions. During the acclimatization period the animals were group housed in Makrolon cages (MIV type, height 18 cm). Diet: Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany). Water: Free access to tap water. Diet, water, bedding and cage enrichment evaluation for contaminants and/or nutrients was performed according to facility standard procedures. There were no findings that could interfere with the study.

Administration / exposure

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

Test Item Preparation The test item was dosed undiluted as delivered by the Sponsor. The test item was kept at room temperature for a maximum of 4 hours prior to dosing. No correction was made for the purity/composition of the test item. Adjustment was made for specific gravity of the test item.TreatmentMethod: Dermal application. The test item was stirred on a magnetic stirrer during application. Clipping: One day before exposure (Day -1) an area of approximately 5x7 cm on the back of each animal was clipped. Application: The test item was applied on an area of approx. 10% of the total body surface, i.e. approx. 25 cm² for males and 18 cm² for females. The test item was held in contact with the skin with a dressing, consisting of a surgical gauze patch (Surgy 1D), successively covered with aluminum foil and Coban elastic bandage. A piece of Micropore tapewas additionally used for fixation of the bandages in females only.

Duration of exposure:

24 hours, after which dressings were removed and the skin cleaned of residual test item using tap water.

Doses:

Frequency: Single dosage, on Day 1. Dose level (volume): 2000 mg/kg (2.13 mL/kg) body weight 5000 mg/kg (5.32 mL/kg) body weight Dose volume calculated as dose level (g/kg) / density (g/mL)

No. of animals per sex per dose:

The study was performed in a step wise fashion. Three females were dosed at 2000 mg/kg in the first step. Based on the absence of mortality and clinical signs, five females and five males were dosed at 5000 mg/kg.

Control animals:

not required

Details on study design:

Observations Mortality/Viability: Twice daily. Body weights: Days 1 (pre-administration), 8 and 15. Clinical signs: At periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15. The time of onset, degree and duration were recorded and the symptoms graded according to fixed scales: Maximum grade 4: grading slight (1) to very severe (4) Maximum grade 3: grading slight (1) to severe (3) Maximum grade 1: presence is scored (1). Necropsy: At the end of the observation period, all animals were sacrificed by oxygen/carbon dioxide procedure and subjected to necropsy. Descriptions of all internal macroscopic abnormalities were recorded.

Statistics:

No specified

Results and discussion

Preliminary study:

Three females were dosed at 2000 mg/kg in the first step. Based on the absence of mortality and clinical signs, five females and five males were dosed at 5000 mg/kg.

Effect levels

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred.

Clinical signs:

At 2000 mg/kg, no systemic clinical signs were noted. Focal erythema was seen on the nose and/or treated skin-area for the animals during the observation period.At 5000 mg/kg chromodacryorrhoea was noted for three males and two females on Day 1. Scales and/or scabs were seen on the back of one male. Erythema and/or scales were seen in the treated skin-area for the animals during the observation period.

Body weight:

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

Gross pathology:

Isolated dark red foci of the thymus were found in one female treated at 5000 mg/kg. No abnormalities were found at macroscopic post mortem examination for any of the other animals.

Other findings:

No further findings noted in the study report.

Any other information on results incl. tables

MORTALITY DATA

TEST DAY HOURS AFTER TREATMENT	1 0	1 2	1 4	2	3	4	5	6	7	8	9	10	11	12	13	14	15
FEMALES 2000 MG/KG	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
MALES 5000 MG/KG	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
FEMALES 5000 MG/KG	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-

 

CLINICAL SIGNS

TEST DAY HOURS AFTER TREATMENT	MAX GRADE	1 0	1 2	1 4	2	3	4	5	6	7	8	9	10	11	12	13	14	15
FEMALES 2000 MG/KG
ANIMAL 1 Skin/fur                Erythema focal (Nose)	(4)	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
ANIMAL 2 Skin/fur                Erythema focal (Nose)                Erythema focal (Treated skin)	(4) (4)	1 -	- -	- -	- 1	- -	- -	- -	- -	- -	- -	- -	- -	- -	- -	- -	- -	- -
ANIMAL 3 Skin/fur                Erythema focal (Nose)                Erythema focal (Treated skin)	(4) (4)	1 -	- -	- -	- 1	- -	- -	- -	- -	- -	- -	- -	- -	- -	- -	- -	- -	- -
MALES 5000 MG/KG
ANIMAL 4 Secretion/excretion                Chromodacryorrhoea (Snout)	(3)	-	1	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-
ANIMAL 5 No clinical signs noted		-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
ANIMAL 6 Secretion/excretion                Chromodacryorrhoea (Snout)	(3)	-	1	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-
ANIMAL 7 Secretion/excretion                Chromodacryorrhoea (Snout)	(3)	-	1	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-
ANIMAL 8 Skin/fur                Scales (Back)                Scabs (Back)	(3) (3)	- -	- -	- -	- -	- -	- -	- -	- -	- -	- -	- -	- 1	- 1	- 1	1 -	1 -	1 -
FEMALES 5000 MG/KG
ANIMAL 9 Skin/fur                General erythema (Treated skin) Secretion/excretion                Chromodacryorrhoea (snout)	(4)   (3)	-   -	-   1	-   1	1   -	-   -	-   -	-   -	-   -	-   -	-   -	-   -	-   -	-   -	-   -	-   -	-   -	-   -
ANIMAL 10 Skin/fur                General erythema (Treated skin)	(4)	-	-	-	1	-	-	-	-	-	-	-	-	-	-	-	-	-
ANIMAL 11 Skin/fur                General erythema (Treated skin)	(4)	-	-	-	1	-	-	-	-	-	-	-	-	-	-	-	-	-
ANIMAL 12 Skin/fur                General erythema (Treated skin)	(4)	-	-	-	1	-	-	-	-	-	-	-	-	-	-	-	-	-
ANIMAL 13 Skin/fur                General erythema (Treated skin)                Scales (Treated skin) Secretion/excretion                Chromodacryorrhoea (Snout)	(4) (3)   (3)	- -   2	- -   2	- -   1	1 -   -	- -   -	- -   -	- 1   -	- 1   -	- -   -	- -   -	- -   -	- -   -	- -   -	- -   -	- -   -	- -   -	- -   -

- = sign not observed

 

BODY WEIGHTS (GRAM)

SEX/DOSE LEVEL	ANIMAL	DAY 1	DAY 8	DAY 15
FEMALES 2000 MG/KG
	1 2 3   MEAN ST.DEV. N	205 194 193   197 7 3	196 199 200   198 2 3	201 207 211   206 5 3
MALES 5000 MG/KG
	4 5 6 7 8   MEAN ST.DEV. N	299 285 274 278 268   281 12 5	312 302 299 282 291   297 11 5	345 327 325 305 318   324 15 5
FEMALES 5000 MG/KG
	9 10 11 12 13   MEAN ST.DEV. N	213 190 197 207 201   202 9 5	220 192 204 214 207   207 11 5	238 204 211 225 213   218 13 5

 

MACROSCOPIC FINDINGS

ANIMAL	ORGAN	FINDING	DAY OF DEATH
FEMALES 2000 MG/KG
1   2   3		No findings   No findings   No findings	Scheduled necropsy Day 15 after treatment Scheduled necropsy Day 15 after treatment Scheduled necropsy Day 15 after treatment
MALES 5000 MG/KG
4   5   6   7   8		No findings noted   No findings noted   No findings noted   No findings noted   No findings noted	Scheduled necropsy Day 15 after treatment Scheduled necropsy Day 15 after treatment Scheduled necropsy Day 15 after treatment Scheduled necropsy Day 15 after treatment Scheduled necropsy Day 15 after treatment
FEMALES 5000 MG/KG
9   10   11   12   13	Thymus	No findings noted   No findings noted   No findings noted   No findings noted   Focus/foci, isolated, dark red	Scheduled necropsy Day 15 after treatment Scheduled necropsy Day 15 after treatment Scheduled necropsy Day 15 after treatment Scheduled necropsy Day 15 after treatment Scheduled necropsy Day 15 after treatment

 

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The dermal LD50 value of MLA-3202 in Wistar rats was established to exceed 5000 mg/kg body weight.

Executive summary:

Assessment of acute dermal toxicity with MLA-3202 in the rat.

 

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

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"

JMAFF Guidelines (2000), including the most recent revisions.

 

Initially, MLA-3202 was administered to three female Wistar rats by a single dermal application at 2000 mg/kg body weight for 24 hours. Based on the results, MLA-3202 was administered to five Wistar rats of each sex by a single dermal application at 5000 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).

 

No mortality occurred.

At 2000 mg/kg, no systemic clinical signs were noted. Focal erythema was seen on the nose and/or treated skin-area for the animals during the observation period.

At 5000 mg/kg chromodacryorrhoea was noted for three males and two females on Day 1. Scales and/or scabs were seen on the back of one male. Erythema and/or scales were seen in the treated skin-area for the animals during the observation period.

The mean body weight gain of all animals during the observation period was within the range expected for rats used in this type of study.

Isolated dark red foci of the thymus were found in one female treated at 5000 mg/kg. No abnormalities were found at macroscopic post mortem examination for any of the other animals.

 

The dermal LD50 value of MLA-3202 in Wistar rats was established to exceed 5000 mg/kg body weight.

Based on these results, MLA-3202 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 (2015) (including all amendments) and Regulation (EC) No 1272/2008 on classification, labelling and packaging of items and mixtures (including all amendments).