Allyl (3-methylbutoxy)acetate

Endpoint summary

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

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)

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Species:

rat

Strain:

Wistar

Sex:

female

Route of administration:

oral: gavage

Vehicle:

other: sesame oil

Doses:

300 mg/Kg body and 2000 mg/Kg body

No. of animals per sex per dose:

3

Control animals:

no

Details on study design:

Test Procedure
A stepwise procedure using 3 female animals per step was used to assess the acute oral toxicity of the test item as described in the OECD guidelines for testing of chemicals, number 423.
In the lack of information about the toxicity of test item, a starting dose of 300 mg/Kg body weight was selected for Step 1. The test procedure was followed as per the attached scheme described in Appendix 1.
All the animals received a single dose of the test item by oral route of administration. Animals were administered with test item after being fasted for 15 h, but with ad libitum water. Feed was given after 3 h and 50 min following administration of test item.
A dose volume of 10 mL/Kg body weight was used. Based on the outcomes of the previous step, further steps were carried out.

Key result

Sex:

female

Dose descriptor:

LD50 cut-off

Effect level:

ca. 500 mg/kg bw

Based on:

test mat.

Mortality:

Mortality and Morbidity
In step 1 and step 2 all the animals were observed for mortality and morbidity for a period of 14 days following the test item administration. Based on mortality and morbidity rate observed at step 1 and step 2, the next steps were started based on the scientific judgement of the Study Director. In step 3 all animals were found dead following the test item administration.

Clinical signs:

Clinical Observation
Clinical observations were performed to look for signs of ill health or overt toxicity during the first 30 minutes and at 1, 2, 3 and 4 h after dose administration on Day 0 and daily during days 1-14 for step 1 and step 2. In step 3 clinical signs were observed at first 30 minutes and at 1, 2, 3 and 4 h after dose administration.
Any abnormalities of appearance or behaviour or other signs of reaction to treatment or ill health were recorded and a detailed individual record was maintained of the clinical condition of each animal.
Observations for signs of toxicity such as changes in skin and fur, eyes and mucous membranes, respiratory, circulatory, autonomic and central nervous systems; and somatomotor activity and behaviour pattern were observed. Attention was also directed to observe for tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.
No animals were found in a moribund condition and showed severe pain or enduring signs of severe distress in step 1 and step 2. In step 3 all animals were found dead after test item administration.

Body weight:

Body Weight
Body weight of each animal were recorded prior to the test item administration (day 0) and on day 7 and day 14 for step 1 and step 2.

Gross pathology:

Necropsy
All the animals in step 1 and step 2 were sacrificed by CO2 exposure at the end of the 14 days observation period and subjected to gross pathology.

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

The acute oral cut-off LD50 value in Wistar rats was 500 mg/kg/b.w. It is classifed as H302

Executive summary:

Acute oral toxicity potential of the test itemAllyl Amyl Glycolate(Batch No. AAG-TEST 1), supplied byMORAYA GLOBAL LIMITED, was evaluated in a 3-step procedure using 3 female (nulliparous and non-pregnant) Wistar rats for each step.

To administer a constant volume over the range of doses to the animal, solvent solubility trail was performed. The test item was not soluble in distilled water & physiological saline but was soluble inDMSO (Dimethyl sulfoxide) & sesame oil, therefore, desired concentration was achieved by mixing the test item with sesame oil.

In step 3 (2000 mg/kg b.w), toxicity symptoms were exhibited immediately after administration of test item. Animals showed signs like dyspnoea (difficult to breathing), abdominal breathing (breathing by diaphragm observed by greater deflection of abdomen), gasping (wheezing sound), nasal discharges, catatonia (decrease in spontaneous motor activities) and piloerection (rough hair standing up) during 30 mins observation. Between 1 h to 4 h, all animals exhibited tachycardia (increase in heart rate) and tucked up abdomen. At the end of 4^thh, all animals were dead (100% mortality). Necropsy revealed severe congestion in liver, kidney and lungs.

Based on the results obtainedin this study and in line withOECD Guideline for Testing of Chemicals, 423 (Adopted on 17^thDecember 2001) it is concludedthatthe given test itemAllylAmyl Glycolate(Batch No. AAG-TEST 1),supplied byMORAYA GLOBAL LIMITED,falls under ‘Category 4’ according to the Globally Harmonized System (GHS) for the classification of chemicals. The acute oralcut-offLD₅₀value in Wistar rats is 500 mg/kg/b.w.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

500 mg/kg bw

Quality of whole database:

Klimisch 1

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Version / remarks:

adopted 7 September, 2009

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Species:

rat

Strain:

Wistar

Sex:

male/female

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

nose only

Vehicle:

other: DMSO

Mass median aerodynamic diameter (MMAD):

>= 3.41 - <= 3.99 µm

Geometric standard deviation (GSD):

>= 1.93 - <= 2.42

Remark on MMAD/GSD:

Galai CIS-50 Aerosol Particle Size Analyser
Instrument: GALAI CIS-50 particle size analyzer
Manufactured by: Galai Pvt. Ltd., Israel
Principle of measurement: Laser based ‘Time-of-Transition Theory’
Unit of measurement: Aerosol particle size in µm.
Calibration: The instrument was calibrated once every three months using
the standards (Latex Microspheres - Thermo Labs, USA) to check the
performance efficiency and the performance of the instrument was within
the specified range of standards (1- 4 µm ).

Details on inhalation exposure:

The equipment, nose only exposure chamber is made of two stainless steel
cylindrical chambers with 24 exposure ports in 4 tiers (6 ports per tier) to
house up to 24 animals. Volume of the inhalation exposure chamber is 53
liters. The animal restrainers are made of polycarbonate tubes having
facility to trap the faeces for individual exposure tubes.
Inner chamber: Diameter 16 cm x Height 40 cm.
Outer chamber: Diameter 38 cm x Height 40 cm.
The equipment consists of two chambers, i.e. the aerosol inlet chamber and
the aerosol exhaust chamber. The chambers are cylindrical in shape and
placed one next to the other. The chamber has 24 ports into a central
plenum arranged as 4 horizontal rows centred on the sides of the cylindrical
chamber.
Animals are restrained in plexiglass exposure restrainers. The approximate
rat restrainer size for males: length 19.0 cm, diameter 6.0 cm and for
females: length 15.0 cm, diameter 6.0 cm.
These restrainers make an airtight seal with the plenum tube through a
rubber gasket. Within the plenum is a central tube into which the aerosol
enters at each port. Air flow carries the aerosol to the nose of the animals
restrained in position at the open ends of the tubes.
Aerosol not breathed and expired air pass into the plenum chamber (aerosol
exhaust chamber) and from there, via filters, to the exhaust system. The
plexiglass restrainers with the animals are housed in an outer chamber. This
chamber is made up of cylindrical plexiglass cover with an opening to fix
the restrainers to the central plenum. The chamber has temperature and
humidity control devices, and the temperature and the humidity where the
animals are housed is controlled and monitored.

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

ca. 4 h

Concentrations:

Based on the pre-study results, the following doses were selected for the main study.
G1 : Dimethyl sulphoxide with an injection rate of 0.4 mL/minute.
G2 : Test item concentration of 7% w/v in dimethyl sulphoxide with an injection rate of 0.4 mL/minute
G3 : Test item concentration of 10% w/v in dimethyl sulphoxide with an injection rate of 0.4 mL/minute
G4 : Test item concentration of 13% w/v in dimethyl sulphoxide with an injection rate of 0.4 mL/minute

No. of animals per sex per dose:

5 male and 5 female

Control animals:

yes

Sex:

male/female

Dose descriptor:

LC50

Effect level:

ca. 0.46 mg/L air

Based on:

test mat.

95% CL:

>= 0.414 - <= 0.517

Exp. duration:

4 h

Sex:

male

Dose descriptor:

LC50

Effect level:

ca. 0.5 mg/L air

Based on:

test mat.

95% CL:

>= 0.445 - <= 0.556

Exp. duration:

4 h

Key result

Sex:

female

Dose descriptor:

LC50

Effect level:

ca. 0.43 mg/L air

Based on:

test mat.

95% CL:

>= 0.386 - <= 0.482

Exp. duration:

4 h

Mortality:

G1: 0
G2: 2 ( 1 female and 1 male) on day 2.
G3: 5 ( 3 female and 2 male) on day 2.
G4: 3 ( 2 female and 1 male) on day 1
G4: 5 ( 2 female and 3 male) on day 2

Clinical signs:

other: The rats were observed for pre-terminal deaths four times during day 1 (at hourly intervals during the exposure) and twice after release of rats from the restrainers for clinical signs of toxicity / pre-terminal deaths and once daily during days 2-15. All

Body weight:

Body weights were recorded once during the acclimatization period and on day 1 (pre-exposure), 2, 4 (3 days post exposure), 8 (7 days post exposure) and 15 (14 days post exposure) and at death.
G1: The body weights of all the rats increased through the observation
period.
G2: The body weights of the all the surviving rats decreased on days 2, 4
when compared to their initial weight however all these rats gained weight
on day 8 and at terminal weighing .The pre-terminally dead rats showed
decrease in weight when compared to their initial weight.
G3 and G4: The body weight of the surviving rats decreased on days 2, 4, 8
when compared to their initial weight however rats gained weight at
terminal weighing. The pre-terminally dead rats showed decrease in weight
when compared to their initial weight.

Gross pathology:

Necropsy
At the end of the observation period, all surviving rats were euthanised using isoflurane anesthesia and subjected to detailed necropsy by an experienced prosector. The pre-terminally dead animals were also subjected to detailed necropsy.
G1: No abnormality was detected in any of the rats at necropsy.
G2 and G3: No abnormality was detected in the pre-terminally dead and
surviving rats at necropsy.
G4: Lung congestion was observed in pre-terminally dead rats (Rm9083,
Rm9086 and Rm9090). No abnormality was detected in rest of the preterminally dead and surviving rats at necropsy.

Interpretation of results:

Category 2 based on GHS criteria

Conclusions:

LC 50 ( male /female) is : 0.46 mg/L of chamber air. It is classifed as H330 Category 2

Executive summary:

The acute inhalation toxicity study of Allyl Amyl Glycolate was conducted
in male and female Wistar rats with one vehicle control (G1) and three
treatment groups by nose only exposure using 7%, 10% and 13% w/v
aerosol of the test item in dimethylsulphoxide, to three groups of rats (G2,
G3 and G4) respectively. The aerosol was generated by a glass atomizer
with an injection rate of 0.4 mL/min, with 1.4 kg/cm2of atomizer pressure.
The rats housed in special rat restrainers were exposed to the test item
aerosol for four hours in an inhalation exposure chamber (dynamic state) for
the respective groups. The post-treatment observation period was 14 days.
The aerosol sampled from the inhalation chamber for particle
size analysis showed a mean aerosol particle size of 1.71 ± 1.18,
1.28 ± 1.05, 1.26 ± 0.99 and 1.31 ± 1.00 micrometers for G1, G2, G3 and
G4 groups respectively. The particle size was measured by using online
particle size analyser (Galai Particle size analyser) instead of normal
Cascade Impacter. So, the particle size is reported as Mean aerosol particle
size instead of Mass Median Aerodynamic Diameter (MMAD).
The generated aerosol particle distribution was uniform throughout the
exposure period of four hours. The geometric standard deviation (GSD)
value of the test item is 2.36, 2.22, 1.93 and 2.42 for G1, G2, G3 and G4
groups respectively, which were greater than 1.2, which indicates that the
aerosol is polydisperse. The fraction of aerosol with particle size of 3.99,
3.97, 3.41 and 3.91 micrometer is less than 95%, 97%, 97% and 97% in
G1,G2, G3 and G4 groups respectively this fraction is considered to be
inhalable up to the alveolar region of the lungs.
The analytically determined average concentrations of Allyl Amyl
Glycolate were 0.25, 0.47 and 0.84 mg per liter of chamber air for G2, G3
and G4 groups respectively.
Clinical signs like, clear nasal discharge, slight / moderate ataxia,
slight tremors/ moderate tremors, slight / moderate salivation, dyspnoea,
hypoactivity, and slight piloerection were observed in treated rats.
Pre-terminal death of 20%, 50% and 80% occurred in G2, G3 and G4 groups
respectively without sex preference. There was reduction in body weight
was observed in all surviving treatment group rats of G2 on days 2, 4 when
compared to their initial weight however all these rats gained weight on day
8 and at terminal weighing. There was reduction in body weight was
observed in all surviving treatment group rats of G3 and G4 on days 2, 4 and
8 when compared to their initial weight, however gained bodyweight at
terminal weighing when compared to the initial weight. The reduction in
body weight was observed in all pre-terminally dead rats.

As there were pre-terminal dead rats no abnormality and lung congestion
was detected. No abnormality was detected in any of the surviving rats
during necropsy.

LC 50 ( male /female) is : 0.46 mg/L of chamber air = 460 mg/m³ of air

LC 50 ( female) is : 0.43 mg/L of chamber air = 430 mg/m³ of air

LC 50 ( male) is : 0.5 mg/L of chamber air = 500 mg/m³ of air

It is classifed as H330 Category 2

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LC50

Value:

430 mg/m³

Quality of whole database:

Klimisch 1

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Version / remarks:

Adopted on 09 October 2017

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

yes

Species:

rat

Strain:

other: Rattus norvegicus

Sex:

female

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

A range finding study using 1 female rat at dose rate of 200 mg/Kg b.w. was carried out in order to establish the dose levels for the main study. Seventeen hours prior to the treatment, around 10% dorsal skin area of each rat was clipped free of hair, without any abrasion. The appropriate amount of the test item was applied uniformly over the fur clipped area of each rat.

After the application, the test item was held in contact with the skin for a period of
24 hours, using a gauze dressing (Make: Liv Medica Products Pvt Ltd; Batch No: S0360617; Expiry: May 2022) and bandaged with non-irritating adhesive tape (Make: 3M India Limited; Batch No: R06160302; Expiry: June 2021). After 24 hours, the residual test item was wiped gently from the skin using cotton soaked in distilled water. Animals were individually caged and neck collar was used to prevent the ingestion of the test item from the application site. No mortality was observed over a period of 72 hours in the range finding study at 200 mg/Kg b.w., 1000 mg/Kg b.w and 2000 mg/Kg b.w. and therefore, main study using 2 female rats at the dose of 2000 mg/Kg b.w. was conducted.

Duration of exposure:

24h

Doses:

Range finding: 200, 1000, 2000 mg/Kg b.w
Main study: 2000 mg/Kg b.w

No. of animals per sex per dose:

range finding: 1 female / dose
main study: 2 females / dose

Control animals:

not required

Details on study design:

Step Study Dose (mg/Kg) Number of animals Number of moribund or dead animals Subsequent action
1 Range finding study 200 1 female 0 Proceeded to Step 2
2 Range finding study 1000 1 female 0 Proceeded to Step 3
3 Range finding study 2000 1 female 0 Proceeded to Step 4
4 Main Study 2000 2 females 0 Further testing not required
Animals were observed at 25 min after application and 45 min, 1 h, 2h, 3h, 4h, 5 h, 6h, 7 h, 8h, 9h, 10h, 13 h, 16 h, 19 h, 22h, 24 h and periodically for a total of 14 days. In addition to that the skin reactions were visually scored at 24 h, 48 h and 72 h after removal of test chemical using Draize criteria.

Statistics:

na

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality or morbidity were observed in any of the animals used in this study.

Clinical signs:

In range finding study (200 mg/kg and 1000 mg/Kg), all animals showed signs of nasal irritation immediately after application. Animals were normal after 5 h of application. At 24 h observation, all animals were found normal.
At 2000mg/kg b.w dose, both in range findings and main study, all the animals showed signs of nasal irritation immediately after application and has staggering gait from 1 h to 2 h. Animals were dull, depressed with decreased motor activity from 2 h to 6 h. Animals became normal after 7th h. At 24 h observation, all animals were found normal.

Body weight:

All the animals showed an increase in body weight at the end of the experiment.

Gross pathology:

No gross and histopathological examination were found necessary in this study.

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the results obtained and in line with OECD 402 the acute dermal LD50 of the given test item Allyl Amyl Glycolate (Batch No. AAG-TEST 1) is greater than 2000 mg/Kg b.w.
CLP criteria not met.

Executive summary:

Based on the results obtained and in line with OECD 402 the acute dermal LD50 of the given test item Allyl Amyl Glycolate (Batch No. AAG-TEST 1) is greater than 2000 mg/Kg b.w.

CLP criteria not met.

Acute dermal toxicity of the test itemAllylAmyl Glycolate(Batch No. AAG-TEST 1), following single dermal application was evaluated in female (nulliparous and non-pregnant) Wistar rats.

Since the test item is a colourless liquid it was used as such without any dilution. In order to establish the dose levels for the main study, a range finding study was initiated using 1 female rat at the dose of 200 mg/Kg b.w. The test item was applied uniformly over the fur clipped dorsal area of the rat (10% of the total body surface area). After application, the test item was held in contact with the skin with gauze dressing which was wrapped with non-irritating tape for a period of 24 hours. Animals were maintained in an individual cage with appropriate gauze dressing and neck collar in order to avoid oral ingestion of the test chemical from the test sites.After 24 hours, the residual test item was wiped gently from the skin using distilled water soaked cotton. Since no mortality was observed in 200 mg/kg b.w. dose, based on OECD 402, a higher dose of 1000 mg/kg b.w. & 2000 mg/kg b.w. was evaluated using the same procedure as before. However, a period of 72 h was allowed between the testing of each animal.

There was no mortality in the range finding study at the dose levels of 200 mg/kg b.w, 1000 mg/kg b.w and 2000 mg kg b.w and therefore, main study using 2 female rats at the dose of 2000 mg/Kg b.w. was conducted.

Step	Study	Dose (mg/Kg)	Number of animals	Number of moribund or dead animals	Subsequent action
1	Range finding study	200	1 female	0	Proceeded to Step 2
2	Range finding study	1000	1 female	0	Proceeded to Step 3
3	Range finding study	2000	1 female	0	Proceeded to Step 4
4	Main Study	2000	2 females	0	Further testing not required

Animals were observed at 25 min after application and 45 min, 1 h, 2h, 3h, 4h, 5 h, 6h, 7 h, 8h, 9h, 10h, 13 h, 16 h, 19 h, 22h, 24 h and periodically for a total of 14 days. In addition to that the skin reactions were visually scored at 24 h, 48 h and 72 h after removal of test chemical using Draize criteria. The mean score of skin reactions are follows,

Animal number	Range finding study			Main study
	1	2	3	4	5
	T	T	T	T	T
Erythema and Eschar formation	0	0	0	0	0
Oedema formation	0	0	0	0	0

   T: Treated

In range finding study (200 mg/kg and 1000 mg/Kg), all animals showed signs of nasal irritation immediately after application. Animals were normal after 5 h of application. At 24 h observation, all animals were found normal.

At 2000mg/kg b.w dose, both in range findings and main study, all the animals showed signs of nasal irritation immediately after application and has staggering gait from 1 h to 2 h. Animals were dull, depressed with decreased motor activity from 2 h to 6 h. Animals became normal after 7^thh. At 24 h observation, all animals were found normal.

No mortality and morbiditywereobserved in any of the animals used in this study.
All the animals showed anincreasein body weight at the end of the experiment.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

Based on the results obtained and in line with OECD 402 the acute dermal LD50 of the given test item Allyl Amyl Glycolate (Batch No. AAG-TEST 1) is greater than 2000 mg/Kg b.w. CLP criteria not met.