Reaction mass of (9-acetoxy-3,8,10-triethyl-7,8,10-trimethyl-1,5-dioxa-9-azaspiro[5.5]undec-3-yl)methyl palmitate and (9-acetoxy-3,8,10-triethyl-7,8,10-trimethyl-1,5-dioxa-9-azaspiro[5.5]undec-3-yl)methyl stearate

Endpoint summary

Description of key information

The substance is virtually non-toxic after a single ingestion and after single dermal exposure.
Oral: LD50 = >2000 mg/kg bw (OECD 423)
Dermal: LD50 = >2000 mg/kg bw (OECD 402)

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:

2003

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Species:

rat

Strain:

other: HanBrI: Wist (SPF)

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: RCC Ltd, Laboratory Animal Services CH-4414 Fullinsdorf / Switzerland
- Age at study initiation: 12-13 weeks
- Weight at study initiation: 170 - 195 g
- Fasting period before study: 17 hours
- Housing: Groups of three in Makrolon type-4 cages with wire mesh tops and standard softwood bedding ('Lignocel' Schill AG, CH-4132 Muttenz/Switzerland)
- Diet: Pelleted standard Provimi Kliba 3433 rat/mouse maintenance diet, batch no. 84/02 (Provimi Kliba AG, CH-4303 Kaiseraugst/Switzerland) ad libitum.
- Water: Community tap water from Fiillinsdorf ad libitum.
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 30-70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

polyethylene glycol

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 0.2 g/mL
- Amount of vehicle (if gavage): 10 ml/kg body weight
- Justification for choice of vehicle: The vehicle was chosen after a non-GLP solubility trial which was performed before the study initiation date.
- Lot/batch no. : 442989/154502013

MAXIMUM DOSE VOLUME APPLIED: 10 ml/kg body weight

DOSAGE PREPARATION: The test item was weighed Into a tared glass beaker on a suitable precision balance and the vehicle added (weight:volume). Homogeneity of the test item in the vehicle was maintained during administration using a magnetic stirrer.

Doses:

2000 mg/kg

No. of animals per sex per dose:

2 x 3 females

Control animals:

no

Details on study design:

- Duration of observation period following administration: 15 days
- Frequency of observations: Daily during acclimatization and at approximately 1, 2, 3 and 5 hours after administration on test day 1. During days 2-15 twice daily for viability and once daily for clinical signs.
- Frequency of weighing: On test days 1 (prior to administration), 8 and 15
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, viability

Statistics:

No statistical analysis was used.

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Mortality:

All animals survived until the end of the study period.

Clinical signs:

other: No clinical signs were observed during the course of the study.

Gross pathology:

No macroscopic findings were recorded at necropsy.

Body weights (g)

Dose (mg/kg)	Animal No.	Sex	Day 1(treatment)	Day 8	Day 15
2000	1	F	185.0	193.3	197.4
	2	F	170.6	185.4	193.0
	3	F	178.4	189.0	200.4
2000	4	F	181.1	188.3	187.4
	5	F	177.6	191.4	196.6
	6	F	194.7	200.5	205.6

Interpretation of results:

practically nontoxic

Conclusions:

The acute oral LD50 of the test substance in female rats is greater than 2000 mg/kg body weight.

Executive summary:

Six female HanBrI: WIST (SPF) rats were treated with the test substance by oral gavage administration at a dosage of 2000 mg/kg body weight. The test item was diluted in vehicle (PEG 300) at a concentration of 0.2 g/mL and administered at a volume dosage of 10mL/kg. All animals survived until the end of the study period. No clinical signs were observed during the course of the study. One female showed a loss of body weight (0.5 %) between test day 8 and the end of the observation period. The body weight of the other animals was within the range commonly recorded for this strain and age. No macroscopic findings were recorded at necropsy.

 

In conclusion, the median lethal dose of the test substance after single oral administration to female rats, observed over a period of 14 days is greater than 2000 mg/kg body weight.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Acute toxicity: via inhalation route

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:

2003

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

other: HanBrI: WIST (SPF)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: RCC Ltd, Laboratory Animal Services CH-4414 Fullinsdorf / Switzerland
- Age at study initiation: Males: 9-10 weeks; Females: 12-13 weeks
- Weight at study initiation: Males: 235-275 g; Females: 180-200 g
- Housing: During acclimatization in groups of five per sex in Makrolon type-4 cages with standard softwood bedding. Individually in Makrolon type-3 cages with standard softwood bedding ("Lignocel", Schill AG, CH-4132 Muttenz) during treatment and observation.
- Diet: Pelleted standard Provimi Kliba 3433 rat/mouse maintenance diet, batch no. 84/02 (Provimi Kliba AG, CH-4303 Kaiseraugst/ Switzerland) ad libitum.
- Water: Community tap water from FQllinsdorf acf libitum
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 10-70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: the backs of the animals
- % coverage: 10
- Type of wrap if used: gauze patch with a semi-occlusive dressing fixed with an elastic adhesive bandage.

REMOVAL OF TEST SUBSTANCE
- Washing: with lukewarm tap water and dried with disposable paper towels
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2g/kg bodyweight

Duration of exposure:

24 hours

Doses:

2000 mg/kg

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 15 days
- Frequency of observations: Daily during acclimatization and at approximately 1, 2, 3 and 5 hours after administration on test day 1. During days 2-15 twice daily for viability and once daily for clinical signs.
- Frequency of weighing: On test days 1 (prior to administration), 8 and 15
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, viability

Statistics:

No statistical analysis was used.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Mortality:

No deaths occurred during the study.

Clinical signs:

other: No systemic or local signs of toxicity were observed during the study period.

Gross pathology:

No macroscopic findings were observed at necropsy.

Individual body weights in grams

Sex / dose	Animal No.	Day of treatment	Day 8	Day 15
male / 2000 mg/kg	1	236.4	249.3	267.7
	2	269.3	285	298.6
	3	271.5	29.38	318.8
	4	243.4	266.0	287.2
	5	254.4	280.3	301.1
female / 2000 mg/kg	1	180.5	184.8	182.1
	2	186.6	194.8	197.7
	3	186.9	185.2	197.1
	4	194.5	201.4	211.3
	5	197.0	208.4	216.6

Interpretation of results:

practically nontoxic

Conclusions:

The acute dermal LD50 of the test substance in rats of both sexes is greater than 2000 mg/kg body weight.

Executive summary:

Five male and five female HanBrI: WIST (SPF) rats were treated with the test substance at 2000 mg/kg by dermal application. The test item was applied undiluted as delivered from the sponsor at an amount dosage of 2 g/kg. The application period was 24 hours. No deaths occurred during the study. No clinical signs were observed during the observation period. One female animal showed a loss of body weight (0.9 %) one week after treatment. It recovered between test day 8 and the end of the observation period. Another female animal lost body weight (1.5 %) from test day 8 to 15. The body weight of the remaining animals was within the range commonly recorded for this strain and age. No macroscopic findings were observed at necropsy.

In conclusion, the median lethal dose of the test substance after single dermal administration to rats of both sexes, observed over a period of 14 days is greater than 2000 mg/kg body weight.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Additional information

According to regulation (EC) No 1907/2006, Annex XI, paragraph 1.5., substances whose physicochemical, toxicological and ecotoxicological properties are likely to be similar or follow a regular pattern as a result of structural similarity may be considered as a group, or "category" of substances. Environmental effects or environmental fate may be predicted from data for reference substance(s) within the group by interpolation to other substances in the group (read-across approach).

The source (CAS 376588-17-9) and target (EC 936-831-9) substance share identical structural elements, the only difference being the length of the carbon chain at the ester function. The source chemical is esterified with stearic acid (C18), whereas the target compound is a mixture of stearate (C18) and palmitate (C16), with the stearate version accounting for about 60% of the chemicals present in the target compound. Therefore, 60% of the target compound are identical with the source chemical and the remaining molecules of the target compound are nearly identical with the only difference being the chain length at the ester group. It is expected that the structural difference between the octadecanoate and the hexadecanoate does not affect the physico-chemical properties, the toxicological and ecotoxicologocal profile as well as the environmental fate. Both substances are characterized by similar values for water solubility, vapor pressure and log POW. Furthermore, it can be assumed that the degradation products of CAS 376588-17-9 are very similar to the degradation products of the mixture containing the octadecanoate and the hexadecanoate. In addition, both substances triggered comparable (eco)toxicological effects.  Therefore read-across to CAS 376588-17-9 is scientifically justified for physico-chemical properties, the toxicological and ecotoxicological profile as well as environmental fate. For more information on the read-across approach, please refer to the Chapter 13 (attached read-across justification).

Acute toxicity was evaluated in two GLP compliant guideline studies according to OECD 423 (RCC, 848423, 2003) and OECD 402 (RCC, 848424, 2003) with the source substance. According to the analogues approach, no additional information is needed to conclude classification and labelling. The test substance was considered as not acute toxic after oral and dermal application.

Justification for classification or non-classification

Classification, Labeling, and Packaging Regulation (EC) No. 1272/2008

The available experimental test data are reliable and suitable for the purpose of classification under Regulation (EC) No.1272/2008. Based on the data, classification for acute toxicity is not warranted under Regulation (EC) No.1272/2008.