Merginol 901

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

January 21 - February 18, 2021

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2021

Report date:

2021

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

December 17, 2001

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

LUBW Landesanstalt für Umwelt Baden-Württemberg, Griesbachstraße 1, 76185 Karlsruhe

Test type:

acute toxic class method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Fatty acids, C16-18 and C18-unsatd., epoxidized, Me esters, oligomeric reaction products with trimethylolpropane

Molecular formula:

Not applicable (UVCB substance)

IUPAC Name:

Fatty acids, C16-18 and C18-unsatd., epoxidized, Me esters, oligomeric reaction products with trimethylolpropane

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Batch number of test material: 0019913739
- Test item No.: 12/0028-3
- Expiry date: November 01, 2021
- Purity: ~ 96%; Mn=760 g/mol; Mw: 990 g/mol; D = Mw/Mn = 1.3; For details see GLP-characterization No. 20L00189
- Physical state / color: Liquid, viscous / yellowish, clear

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature
- Homogeneity: The test item was homogeneous by visual inspection.
- Storage stability: The stability of the test item under storage conditions over the study period was guaranteed by the sponsor, and the sponsor holds this responsibility.

- Analysis of feed: The feed used in the study was assayed for chemical and microbial contaminants by the manufacturer in regular intervals (several times a year). Results are archived by Bioassay.
- Analysis of drinking water: The drinking water was regularly assayed for contaminants by the municipal authorities of Heidelberg. The German Drinking Water Regulation of Dec. 5, 1990 served as the guideline for maximum tolerable contaminants.
- Bedding and enrichment analysis: The bedding and enrichment were regularly assayed for contaminants (chlorinated hydrocarbons and heavy metals).

FORM AS APPLIED IN THE TEST
- Test item homogenization until the end of each administration period:
The homogeneity of the test item during administration was ensured by stirring with a magnetic stirrer.
- Form of administration: Undiluted

OTHER SPECIFICS
- Density [g/mL]: 0.984 (determined by Bioassay Laboratories)

Test animals

Species:

rat

Strain:

Wistar

Remarks:

Crl:WI (Han) SPF

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Wiga GmbH, Germany
- Females nulliparous and non-pregnant: yes
- Age at study initiation: Young adult animals (female animals approx. 10-11 weeks)
- Weight at study initiation: Animals of comparable weight (± 20% of the mean weight, actual weights see Table 5)
- Housing: Makrolon cage, type III; Single housing
- Fasting period: Feed was withdrawn from the animals at least 16 hours before administration, but water was available ad libitum.
- Diet: ad libitum; LASQCdiet® Rod16, HiHyg, LASvendi (Altromin, 32791 Lage, Germany)
- Water: ad libitum; Tap water
- Bedding: H 15005-29; Ssniff, Spezialdiäten GmbH (Experimental Animal Diets Inc., 59494 Soest, Germany)
- Enrichment: Wooden gnawing blocks (Type NGM E-022); ABEDD® LAB & VET Service GmbH, Hasnerstraße 84/6; 1160 Wien – Austria
- Acclimation period: At least 5 days before the beginning of the experimental phase; during the acclimatization period, the animals were accustomed to the environmental conditions of the study and to the diet.
- Identification: Individual identification by cage cards and tail marking.

ENVIRONMENTAL CONDITIONS
- Temperature: 22°C +/- 3°C
- Humidity: 30 – 70 %
- Air changes (per hr): Approx. 10
- Photoperiod (hrs dark/hrs light): 12h/12h (6.00a.m. – 6.00p.m./ 6.00p.m.–6.00a.m.)

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

3 (see table 1)

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Time of day of administration: In the morning
- Frequency of weighing: Individual body weights shortly before administration (day 0), weekly thereafter and on the last day of observation.
- Clinical observations: Clinical signs for each animal were recorded several times on the day of administration and at least once during each workday thereafter.
- Mortality: A check for any dead or moribund animals was made at least once each workday.
- Necropsy of survivors performed: yes; Necropsy with gross-pathology examination was performed on the last day of the observation period after sacrifice by CO2-inhalation in a chamber with gradually increasing concentrations.

Statistics:

Calculations were performed using Microsoft Excel 2010 and checked with a calculator.

Results and discussion

Effect levels

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: no mortality occured

Mortality:

No mortality occurred in both test groups dosed with 2000 mg/kg bw.

Clinical signs:

other: In the first 2000 mg/kg bw. test group impaired general state and piloerection were observed in all animals from hour 1 or 2 until hour 5, while diarrhea was seen in two of these animals at hour 3 or from hour 2 until hour 4. In addition, dyspnoea and cow

Gross pathology:

There were no macroscopic pathological findings in any animal sacrificed at the end of the observation period.

Any other information on results incl. tables

Table 2: MORTALITY

Mortality
Dose (mg/kg bw):	2000	2000
Sex:	female	female
Administration:	1	2
No. of animals:	3	3
Mortality (animals):	No mortality	No mortality

Table 3: MAXIMUM INCIDENCE OF CLINICAL SIGNS I

Dose (mg/kg bw):		2000
Sex:		female
Administration:		1
No. of animals:		3
Animal No.:	R 142	R 143	R 144
Abnormalities:
Impaired general state:	h2 - h5	h1 - h5	h2 - h5
Dyspnoea:	-	h3 - h4	-
Piloerection:	h2 - h5	h1 - h5	h2 - h5
Diarrhea:	-	h3	h2 - h4
Cowering position:	-	h3 - h4	-

Table 4: MAXIMUM INCIDENCE OF CLINICAL SIGNS II

Dose (mg/kg bw):		2000
Sex:		female
Administration:		2
No. of animals:		3
Animal No.:	R 148	R 149	R 150
Abnormalities:
Impaired general state:	-	h3 - h4	h3 - h4
Piloerection:	-	h3 - h4	h3 - h4
Diarrhea:	-	h3	h3

Table 5: BODY WEIGHTS

Individual body weight changes
Dose (mg/kg bw):	2000					2000
Administration:	1					2
Animal No.:	R	R	R	Mean weight	Standard- deviation	R	R	R	Mean weight	Standard- deviation
	142	143	144			148	149	150
Body weight at study day (g):
0	194	192	196	194.0	2.00	188	184	190	187.3	3.06
7	219	202	216	212.3	9.07	208	206	210	208.0	2.00
14	228	204	223	218.3	12.66	219	216	220	218.3	2.08

Table 6: GROSS PATHOLOGY

Gross Pathology
Dose (mg/kg bw):	2000			2000
Administration:	1			2
No. of animals:	3			3
Animal No.:	R 142	R 143	R 144	R 148	R 149	R 150
Macroscopic pathologic abnormalities:	-	-	-	-	-	-

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of this study the median lethal dose of the test substance after oral administration was assessed to be greater than 2000 mg/kg bw in rats.

Executive summary:

In an acute oral toxicity study performed according to the Acute Toxic Class Method, a dose of 2000 mg/kg bw of the undiluted test item was administered by gavage to two test groups of three fasted Wistar rats each (6 females).
The following test substance-related clinical observations were recorded, clinical signs occurred within the first 5 hours after administration:

2000 mg/kg (first test group):
- No mortality occurred
- Impaired general state in all animals
- Dyspnoea in one animal
- Piloerection in all animals
- Diarrhea in two animals
- Cowering position in one animal

2000 mg/kg (second test group):
- No mortality occurred
- Impaired general state in two animals
- Piloerection in two animals
- Diarrhea in two animals

All animals gained weight in a normal range throughout the study period.
There were no macroscopic pathological findings in any animal sacrificed at the end of the observation period (6 females).
The acute oral LD50 was assessed to be LD50, oral, rat > 2000 mg/kg bw