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Description of key information

The acute oral median lethal dose (LD50 cut- off value) of Fatty acids, C16-18 (even numbered), manganese(II) salts in Wistar rats was found to be 5000 mg/kg bw. The LD50 for acute dermal toxicity was found to be > 2000 mg/kg bw.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
5 September 2018 (Study Initialtion) to 30 October 2019 (Experiment Completion)
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
yes
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source of test material: Supplied by Nitika Pharmaceutical Specialities Pvt. Ltd.
- Lot/batch No.of test material: MNST9H122A
- Expiration date of the lot/batch: May 2023
- Purity test (release) date: 12 June 2018 (CoA)

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature (Ambient). Container kept tightly closed in in a cool and well ventilated place
- Stability under test conditions: Assumed stable for the duration of the study
- Solubility and stability of the test substance in the solvent/vehicle: Formed a homogenous suspension in corn oil. Assumed stable for the duration of the study
- Reactivity of the test substance with the solvent/vehicle: None

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: Prepared as a homogenous suspension in corn oil prior to dosing
- Preliminary purification step (if any): None

FORM AS APPLIED IN THE TEST (if different from that of starting material) : Test item administered as a homogenous suspension in corn oil
Species:
rat
Strain:
Wistar
Remarks:
Han Tac:WH
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Vivo BioTech Ltd. Pregnapur, Telangana, India
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 9 to 11 weeks
- Weight at study initiation: 158.2g to 174.1 (groups 1 & 2, 300 mg/kg bw), 175.9 to 185.0g (groups 3 & 4, 2000 mg/kg bw)
- Fasting period before study: Rats were fasted overnight prior to dosing and for three hours post-dose
- Housing: Polypropylene rat cages covered with a stainless steel grid top. Autoclaved clean rice husk as bedding material. Wooden chew blocks as enrichment material. Three rats per cage
- Diet (e.g. ad libitum): yes (with the exception of overnight fasting and fasting threee hours post dose)
- Water (e.g. ad libitum): yes
- Acclimation period: 6 to 12 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 to 23°C
- Humidity (%): 58 to 66%
- Air changes (per hr): Minimum of 15 air changes/hour
- Photoperiod (hrs dark / hrs light): 12 hours artificial light and 12 hours darkness, light hours being 06:00 h – 18:00 h (maintained through an automatic timer)

IN-LIFE DATES: From: To: 26 September 2018 (Experimental start) to 30 October 2018 (Experimental completion)
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 30 mg/mL and 200 mg/mL (i.e. 300 and 2000 mg/kg bw at a dose volume of 10 mL/kg bw)
- Amount of vehicle (if gavage): 1.58 to 1.74 mL (300 mg/kg bw groups) and 1.76 to 1.85 mL (2000 mg/kg bw groups)
- Justification for choice of vehicle: In accordence with OECD 423, as the test item was insoluble in water, corn oil was selected as an acceptable vehicle as it formed a homogenous dosable suspension

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: As no toxicological information was available on the test item, in accordance with OECD 423 a starting dose of 300 mg/kg bw was selected as the initial test dose
Doses:
300 mg/kg bw (i.e. 30 mg/mL at 10 mL/kg bw)
2000 mg/kg bw (i.e. 200 mg/mL at 10 mL/kg bw)
No. of animals per sex per dose:
6 female rats per dose (3 females/set)
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Rats were observed for signs of toxicity and mortality at 0.5, 1, 2, 3, 4 and 5 h post-administration on the day of dosing. Rats were observed twice a day for morbidity and mortality for a period of 14 days following oral dosing. Clinical signs were recorded once a day. Individual body weight was recorded prior to dosing on day 0 and on days 7 and 14.

- Necropsy of survivors performed: yes, at the end of the 14 day observation period, all rats were euthanised by carbon dioxide asphyxiation and were subject to gross pathological examination, consisting of external examination and opening of the abdominal and thoracic cavities.
Key result
Sex:
female
Dose descriptor:
LD50 cut-off
Effect level:
>= 5 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality was observed in rats treated with 300 or 2000 mg Fatty acids, C16-18 (even numbered), manganese(II) salts/kg bw
Clinical signs:
No clinical signs were observed in rats treated with 300 or 2000 mg Fatty acids, C16-18 (even numbered), manganese(II) salts/kg bw
Body weight:
Normal gain in body weight was observed in all rats treated with 300 or 2000 mg Fatty acids, C16-18 (even numbered), manganese(II) salts/kg bw
Gross pathology:
Necropsy (Macroscopic Findings)

External examination of terminally sacrificed rats did not reveal any abnormality.
Internal (vIsceral) examination of terminally sacrificed rats did not reveal any abnormality.

In the absence of any pathological lesion in terminally sacrificed rats, it is concluded that the test item did not produce any treatment related effect at the dose levels used.
Interpretation of results:
Category 5 based on GHS criteria
Remarks:
or unclassified
Conclusions:
The acute oral median lethal dose (LD50 cut- off value) of Fatty acids, C16-18 (even numbered), manganese(II) salts in Wistar rats was found to be 5000 mg/kg bw.

Based on results of this study, the classification for Fatty acids, C16-18 (even numbered), manganese(II) salts is as follows:
Globally Harmonized System of Classification and Labelling of Chemicals (GHS 2017) : Category 5 or Unclassified
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
5 000 mg/kg bw
Quality of whole database:
Klimisch 1

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
the study does not need to be conducted because exposure of humans via inhalation is not likely taking into account the vapour pressure of the substance and/or the possibility of exposure to aerosols, particles or droplets of an inhalable size
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:
4 March 2019 (Study Initiation) to XX 2019 (Study Completion)
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
fixed dose procedure
Limit test:
yes
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source of test material: Nitika Pharmaceutical Specialities Pvt. Ltd.
- Lot/batch No.of test material: MNST9H796L
- Expiration date of the lot/batch: August 2023
- Purity test (release) date: September 2018

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature (ambient). Container kept tightly closed in a dry, cool and well ventilated place.
- Stability under test conditions: Assumed stable for the duration of the test
- Solubility and stability of the test substance in the solvent/vehicle: Solubility not applicable. The required amount of test item was applied (moistened with Reverse Osmosis (RO) water) over the clipped dorsolateral thoracic surface of the skin (approximately 7 cm × 5 cm body surface area). The test item contact period with the skin was 24 hours, rats were thereafter observed for a period of 14 days.

- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: None

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: None, test item administered as supplied (moistened with Reverse Osmosis water) before application.


OTHER SPECIFICS:
- measurement of pH, osmolality, and precipitate in the culture medium to which the test chemical is added: The test item pH (1% solution in distilled water at room temperature) was checked and found to be acceptable at 7.07 ± 0.03.
Species:
rat
Strain:
Wistar
Remarks:
RccHan
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Animal Breeding Facility, Jai Research Foundation, India
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 12 to 14 weeks
- Weight at study initiation: Minimum: 253.8, Maximum: 284.3
- Fasting period before study: None
- Housing: Polypropylene rat cages covered with stainless steel grid tops. Autoclaved clean rice husk as bedding material. Wooden blocks were provided as enrichment material.
- Diet: Teklad certified Global High Fiber Rat/Mice Feed manufactured by Envigo, USA. ad libitum
- Water: UV sterilized water filtered through a Reverse Osmosis water filtration system ad libitum
- Acclimation period: 7 to 19 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 to 23 °C
- Humidity (%): 57 to 67%
- Air changes (per hr): Minimum of 15/hour
- Photoperiod (hrs dark / hrs light): The photoperiod was maintained through automatic timer (12 h artificial light and 12 h darkness), light hours being 06:00 – 18:00 h.

IN-LIFE DATES: From: To: 9 March 2019 (Experimental Start) to 11 April 2019 (Experimental Completion)
Type of coverage:
occlusive
Vehicle:
other: moistened prior to application with RO water
Details on dermal exposure:
TEST SITE
- Area of exposure: The fur of each rat was closely clipped from the dorsal area of the trunk using an electric clipper without abrading or damaging the skin. An area greater than 10 percent of the body surface area was clipped approximately 24 h prior to test item application.
- % coverage: An area greater than 10 percent of the body surface area was clipped approximately 24 h prior to test item application.
- Type of wrap if used: The test item was held in contact with the skin using porous gauze dressing (not more than 8 ply) and a non-irritating tape (Medi tape 330, hypo-allergenic surgical tape) throughout the 24 h exposure period to prevent any loss of test item and also to ensure that the rats did not lick or ingest test item.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): At the end of the 24 h exposure period any residual test item was removed using cotton soaked in RO water.
- Time after start of exposure: 24h

TEST MATERIAL
- Amount(s) applied (weight with unit): Based on individual body weights, the required amount of test item was applied (moistened with RO water).
- For solids, paste formed: no

VEHICLE
Not applicable, test item administered as supplied (moistened with RO water) over the clipped dorsolateral thoracic surface of the skin.
Duration of exposure:
24h
Doses:
Range finding study; 200, 1000 and 2000 mg/kg bw
Main study; 2000 mg/kg bw
No. of animals per sex per dose:
1 for each dose range finding concentration
2 for the main study at 2000 mg/kg bw
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Rats were observed for signs of toxicity and mortality within 0.5 hours and at 2, 4 and 6 h post-dermal application on the day of dosing (day 0). Subsequently, rats were observed twice a day for morbidity and mortality for a period of 14 days following dermal application. Rats were observed for erythema and oedema at 24 h, 48 h and 72 h post patch removal. The clinical signs were recorded once a day. Individual body weight was recorded prior to dermal application on day 0 and on days 7 and 14 post dermal application.

- Necropsy of survivors performed: yes, all rats at the end of the 14-day observation period were euthanised by carbon dioxide asphyxiation and subjected to a gross pathological examination consisting of an external examination and opening of abdominal and thoracic cavities.

- Other examinations performed: clinical observations and body weight
Statistics:
None
Preliminary study:
As no toxicological information was available for the test item, a range finding study was conducted. Initially, rat N˚ 1 was dosed at 200 mg Fatty acids, C16-18 (even numbered), iron(III) salts/kg body weight. As no mortality was observed after 72 h exposure, rat N˚ 2 was dosed at 1000 mg Fatty acids, C16-18 (even numbered), iron(III) salts/kg body weight. rat N˚ 3 was dosed at the limit guideline dose of 2000 mg Fatty acids, C16-18 (even numbered), manganese(II) salts/kg body weight.
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
no indication of skin irritation up to the relevant limit dose level
Mortality:
No morbidity and mortality was observed in any rat treated with 200, 1000 or 2000 mg Fatty acids, C16-18 (even numbered), manganese(II) salts/kg bw.
Clinical signs:
No sign of toxicity was observed in any rat treated with 200, 1000 or 2000 mg Fatty acids, C16-18 (even numbered), manganese(II) salts/kg bw.
No erythema or oedema was observed at any 24 h time point interval (24 h, 48 h and 72 h) post patch removal in any rat.
Body weight:
Normal gain in body weight was observed in all rats treated with 200, 1000 or 2000 mg Fatty acids, C16-18 (even numbered), iron(III) salts/kg bw.
Gross pathology:
Necropsy (Macroscopic Findings)
External examination of terminally sacrificed rats did not reveal any abnormality.
Internal (visceral) examination of terminally sacrificed rats did not reveal any abnormality.
Interpretation of results:
GHS criteria not met
Conclusions:
As per the Globally Harmonized System of Classification and Labelling of Chemicals (GHS 2017), if the test item is classified under Category 5 or Unclassified it indicates that the LD50 is established as greater than 2000 mg/kg body weight.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
Klimisch 1

Additional information

Justification for classification or non-classification

The acute oral median lethal dose (LD50 cut- off value) of Fatty acids, C16-18 (even numbered), manganese(II) salts in Wistar rats was found to be 5000 mg/kg bw. The LD50 for acute dermal toxicity was found to be > 2000 mg/kg bw. This substance therefore will not be classified.