Calcium 3,5,5-trimethylhexanoate

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

16 December 2021 - 31 December 2021

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2022

Report date:

2022

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Calcium 3,5,5-trimethylhexanoate

EC Number:

264-731-9

EC Name:

Calcium 3,5,5-trimethylhexanoate

Cas Number:

64216-15-5

Molecular formula:

Ca(C9H18O2)2

IUPAC Name:

calcium bis(3,5,5-trimethylhexanoate)

Test material form:

solid: particulate/powder

Details on test material:

Supplier: Venator Pigments UK Limited
CAS Number: 64216-15-5
Batch/Lot number: VR6-461-15/24
Appearance: white powder
Purity: treated as 100%
Ca content: 11.78%
Expiry date: 05 October 2023

Test animals

Species:

rat

Strain:

Wistar

Remarks:

Han:WIST rats

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Toxi-Coop Zrt., H-1122 Budapest, Magyar Jakobinusok tere 4B
- Hygienic level at supplier: SPF
- Hygienic level during the study: Standard housing conditions
- Number of animals: 6 animals, 3 animals/group
- Sex: Female, nulliparous and non-pregnant animals
- Age of animals at dosing: Young adult rats, approx. 8-9 weeks old
- Body weight range at dosing: 169-194 g. The maximum difference of individual animal weights from the mean of the treatment group did not exceed 20%.
- Acclimatisation period: At least 8 days
- Animal health: Only healthy animals were used for the test. The health status was certified by the Veterinarian.
- Housing: Group caging (3 animals/cage)
- Cage type: T3H polycarbonate
- Bedding and nesting: “SAFE 3/4-S-FASERN” certified wooden chips and “Sizzle pet” nest material were available to animals during the study.
- Enrichment: Animals were housed by group to allow social interaction and with deep wood sawdust bedding to allow digging and other normal rodent activities.
- Animals received standard laboratory rat diet, ad libitum, and tap water from the municipal supply, as for human consumption from drinking bottles designed for rodents, ad libitum.
- The night before treatment, the animals were fasted. Food, but not water, was withheld overnight. Animals were weighed before dosing. Food was replaced 3 hours after the treatment.

ENVIRONMENTAL CONDITIONS
- Lighting period: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
- Temperature: 21 – 23 °C (target: 22 ± 3 °C)
- Relative humidity: 36 – 58 % (target: 30 – 70 %)
- Ventilation: 15-20 air exchanges/hour

IN-LIFE DATES: From 16 December 2021 to 31 December 2021

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

polyethylene glycol

Remarks:

Polyethylene glycol 400 (PEG 400)

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 200 mg/mL
- Amount of vehicle (if gavage): 10 mL/kg bw
- Justification for choice of vehicle: The test item did not dissolve in distilled water. Upon formulation of the test item in aqueous 1% methylcellulose, the test item swelled and formed a dense, stiff mass, which was not compatible with gavage application. Upon formulation of the powdered test item in PEG400 in a mortar, a homogenous suspension was obtained, which if kept under stirring, was sufficiently stable for an application via gavage.
- Lot/batch no.: S8092385
- Manufacturer: Merck
- Expiry date: 31 March 2024

DOSAGE PREPARATION: The test item was freshly formulated in the vehicle at the appropriate concentration (200 mg/mL), in the Pharmacy of NEXTREAT Laboratories on the day of administration. The formulations were stirred with magnetic stirrer up to finishing the treatment.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: As starting dose level for acute toxicity study, a dose of 2000 mg/kg body weight (bw) has been selected based on the data, provided by the Supplier, which indicate that the LD50 value of the test item is between 300 and 2000 mg/kg bw.
- Initially three animals were treated at the starting dose of 2000 mg/kg bw (Group 1). All three animals survived, temporarily showing clinical signs, thus the second group (Group 2) was treated at the same dose level (2000 mg/kg bw). In the second group, two animals behaved similarly to those in the first group, but as one animal showed severe clinical symptoms, after consultation with the Veterinarian it was decided to pre-terminally euthanize it. As per the guideline, this is considered in the interpretation of the test results in the same way as animals that died on test. As no further mortality was observed in this confirmatory dose group, no additional testing was required according to the criteria for termination given in Annex 2d of OECD Guideline No. 423.

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

6 (3 animals per group)

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Clinical observations: Following the end of the dosage, the animals were observed individually once during the first 30 minutes, at 1, 2, 3, 4 and 6 hours after the treatment and once daily for 14 consecutive days thereafter. Individual observations were performed on the skin and fur, eyes and mucous membranes and also respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma. The time of death was recorded as precisely as possible.
- Body weight: The body weight of the animals was recorded on Days 0 (prior to dosing), 7 and 14 (prior to necropsy), with a precision of 1 g.
- Necropsy: Animals were subjected to a necropsy and a macroscopic examination. After examination of the external appearance, the cranial, thoracic and abdominal cavities were opened and the appearance of the tissues and organs were observed. All gross macroscopic changes were recorded for each animal.

Statistics:

The method used was not intended to allow the calculation of a precise LD50 value.

Results and discussion

Effect levels

Effect levels 1

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

>= 2 000 mg/kg bw

Based on:

test mat.

Effect levels 2

Sex:

female

Dose descriptor:

LD50 cut-off

Effect level:

2 500 mg/kg bw

Based on:

test mat.

Mortality:

At the dose level of 2000 mg/kg bw, 1/6 animals was euthanized pre-terminally because of animal welfare reasons on Day 0. The other animals (5/6) survived.

Clinical signs:

other:

Body weight:

lower than 10% body weight loss

Remarks:

There were no effects on body weight or body weight gain that could be attributed to treatment with the test item.

Gross pathology:

In the pre-terminally euthanized animal, diffuse, dark red discoloration of the glandular mucosa of the stomach and diffuse, dark red discoloration of the mucosa of the duodenum and jejunum and was observed. There were no other macroscopic changes seen at the necropsy of the other animals.

Applicant's summary and conclusion

Interpretation of results:

Category 5 based on GHS criteria

Remarks:

The test substance is not classified according to Regulation (EC) No 1272/2008 (CLP)

Conclusions:

Under the conditions of this study, the acute oral LD50 value of the test item Calcium isononanoate was found to be above 2000 mg/kg bw in female Han:WIST rats.
The LD50 cut-off value is 2500 mg/kg bw.
The study result triggers the following classification/labelling:
- Regulation (EC) No 1272/2008 (CLP): Unclassified
- GHS (rev. 7) 2017: Category 5