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

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

25 March 2016 to 17 May 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 423 (Acute Oral toxicity - Acute Toxic Class Method)

Version / remarks:

Organization for Economic Co-operation and Development (OECD), OECD Guidelines for Testing of Chemicals, Section 4, Health Effects. No.423, "Acute Oral Toxicity - Acute Toxic Class Method", 2001.

Deviations:

yes

Remarks:

See "Any other information" for details

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Version / remarks:

Commission Regulation (EC) No 440/2008 Part B: Methods for the Determination of Toxicity and other Health Effects; B1 tris: "Acute Oral Toxicity, Acute Toxic Class Method". Official Journal of the European Union No. L142, May 2008, including the most recent amendments.

Deviations:

yes

Remarks:

See "Any other information" for details

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Version / remarks:

United States Environmental Protection Agency (EPA). Health Effects Test Guidelines, OPPTS 870.1100, Acute Oral Toxicity. Office of Prevention, Pesticides and Toxic Items (7101), EPA 712-C-02-190, 2002.

Deviations:

yes

Remarks:

See "Any other information" for details

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: JMAFF 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:

acute toxic class 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:

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: 9 Females (nulliparous and non-pregnant). Age and body weight: Young adult animals (approx. 8-10 weeks old) were selected. Body weight variation did not exceed +/- 20% of the sex mean. Identification: Earmark and tail mark Health inspection: At least prior to dosing. It was ensured that the animals were healthy and without any abnormality that might have affected the study integrity.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: Group housing of maximally 3 animals per cage in labeled Makrolon cages (MIV 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. 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

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

The toxicity of the test item was assessed by stepwise treatment of groups of females. The first group was treated at a dose level of 2000 mg/kg. The absence or presence of mortality of animals dosed at one step determined the next step, based on the test procedure defined in the guidelines. The onset, duration and severity of the signs of toxicity were taken into account for determination of the time interval between the dose groups.

Doses:

Single dosage on Day 1.

No. of animals per sex per dose:

3 animals per group

Control animals:

no

Details on study design:

Treatment Method: Oral gavage, using plastic feeding tubes. The test item were stirred on a magnetic stirrer during dosing. Fasting: Animals were deprived of food overnight prior to dosing and until 3-4 hours after administration of the test item. Water was available ad libitum. 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. No correction was made for the purity of the test item.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 signs were graded according to fixed scales and the time of onset, degree and duration were recorded: 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 statistical analysis was performed.

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred at 2000 and 5000 mg/kg body weight.

Clinical signs:

Hunched posture was seen for all animals on Day 1 and for one animal dosed at 2000 mg/kg on Days 2 and 3 also. Piloerection was seen for the majority of animals on Day 1. Two females dosed at 5000 mg/kg showed abnormal licking on Day 2.

Body weight:

The body weight gain shown by the animals over the study period was considered to be similar to that expected for normal untreated animals of the same age and strain.

Gross pathology:

Isolated reddish foci on the thymus were found at macroscopic post mortem examination of one female dosed at 2000 mg/kg. Macroscopic post mortem examination of the other animals at termination did not reveal any abnormalities.

Other findings:

Incidental findings included an accessory lobe to the right median lobe of the liver for one female dosed at 2000 mg/kg. This finding is occasionally seen among rats of this age and strain and was therefore considered not related to treatment.

Any other information on results incl. tables

Mortality data

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

 

Clinical signs

TEST DAY	MAX GRADE	1	1	1	2	3	4	5	6	7	8	9	10	11	12	13	14	15
HOURS AFTER TREATMENT		0	2	4
FEMALES 2000 MG/KG
ANIMAL 1 Posture                Hunched posture Skin / fur                Piloerection	(1)   (1)	1   -	1   1	1   1	-   -	-   -	-   -	-   -	-   -	-   -	-   -	-   -	-   -	-   -	-   -	-   -	-   -	-   -
ANIMAL 2 Posture                Hunched posture Skin / fur                Piloerection	(1)   (1)	1   -	1   1	1   1	-   -	-   -	-   -	-   -	-   -	-   -	-   -	-   -	-   -	-   -	-   -	-   -	-   -	-   -
ANIMAL 3 Posture                Hunched posture Skin / fur                Piloerection	(1)   (1)	1   -	1   1	1   1	1   -	1   -	-   -	-   -	-   -	-   -	-   -	-   -	-   -	-   -	-   -	-   -	-   -	-   -
FEMALES 2000 MG/KG
ANIMAL 4 Posture                Hunched posture	(1)	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
ANIMAL 5 Posture                Hunched posture	(1)	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
ANIMAL 6 Posture                Hunched posture	(1)	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
FEMALES 5000 MG/KG
ANIMAL 7 Posture                Hunched posture Skin / fur                Piloerection	(1)   (1)	-   -	1   1	1   1	-   -	-   -	-   -	-   -	-   -	-   -	-   -	-   -	-   -	-   -	-   -	-   -	-   -	-   -
FEMALES 5000 MG/KG
ANIMAL 8 Behaviour                Abnormal licking Posture                Hunched posture Skin / fur                Piloerection	(1)   (1)   (1)	-   -   -	-   1   1	-   -   -	1   -   -	-   -   -	-   -   -	-   -   -	-   -   -	-   -   -	-   -   -	-   -   -	-   -   -	-   -   -	-   -   -	-   -   -	-   -   -	-   -   -
ANIMAL 9 Behaviour                Abnormal licking Posture                Hunched posture Skin / fur                Piloerection	(1)   (1)   (1)	-   -   -	-   1   1	-   -   -	1   -   -	-   -   -	-   -   -	-   -   -	-   -   -	-   -   -	-   -   -	-   -   -	-   -   -	-   -   -	-   -   -	-   -   -	-   -   -	-   -   -

. Observation not performed. Sufficient data was available to warrant the study integrity.

 

Body weights

SEX/DOSE LEVEL	ANIMAL	DAY 1	DAY 8	DAY 15
FEMALES 2000 MG/KG	1 2 3   MEAN ST. DEV. N	150 164 165   160 8 3	171 202 191   188 16 3	193 211 208   204 10 3
FEMALES 2000 MG/KG	4 5 6   MEAN ST. DEV. N	158 167 178   168 10 3	182 197 206   195 12 3	192 206 216   205 12 3
FEMALES 5000 MG/KG	7   MEAN ST. DEV. N	167   167 -- 1	199   199 -- 1	207   207 -- 1
FEMALES 5000 MG/KG	8 9   MEAN ST. DEV. N	174 171   173 2 2	195 190   193 4 2	200 203   202 2 2

 

Macroscopic findings

ANIMAL	ORGAN	FINDING	DAY OF DEATH
FEMALES 2000 MG/KG
1		No findings noted	Scheduled necropsy Day 15 after treatment
2		No findings noted	Scheduled necropsy Day 15 after treatment
3	Liver	Right medial lobe: accessory liver	Scheduled necropsy Day 15 after treatment
FEMALES 2000 MG/KG
4	Thymus	Focus/foci, isolated, reddish	Scheduled necropsy Day 15 after treatment
5		No findings noted	Scheduled necropsy Day 15 after treatment
6		No findings noted	Scheduled necropsy Day 15 after treatment
FEMALES 5000 MG/KG
7		No findings noted	Scheduled necropsy Day 15 after treatment
FEMALES 5000 MG/KG
8		No findings noted	Scheduled necropsy Day 15 after treatment
9		No findings noted	Scheduled necropsy Day 15 after treatment

 

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The oral LD50 value of MLA-3202 in Wistar rats was established to exceed 5000 mg/kg body weight.According to the OECD 423 test guideline, the LD50 cut-off value was considered to exceed 5000 mg/kg body weight.

Executive summary:

The study was performed to assess the acute oral toxicity of MLA-3202 in the rat (Acute Toxic Class Method). 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, B1 tris: “Acute Oral Toxicity, Acute Toxic Class Method”

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

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

 

Initially, MLA-3202 was administered by oral gavage to three female Wistar rats at 2000 mg/kg body weight. In a stepwise procedure, one additional group of three females was dosed at 2000 mg/kg, one female at 5000 mg/kg and one group of two females at 5000 mg/kg body weight. Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice (Day 15).

 

No mortality occurred.

 

Hunched posture was seen for all animals on Day 1 and for one animal dosed at 2000 mg/kg on Days 2 and 3 also. Piloerection was seen for the majority of animals on Day 1. Two females dosed at 5000 mg/kg showed abnormal licking on Day 2.

 

The body weight gain shown by the animals over the study period was considered to be similar to that expected for normal untreated animals of the same age and strain.

 

Isolated reddish foci on the thymus were found at macroscopic post mortem examination of one female dosed at 2000 mg/kg. No other test item related abnormalities were noted in any of the remaining animals.

 

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

 

According to the OECD 423 test guideline, the LD50 cut-off value was considered 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 oral 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).