Phenethyl phenylacetate

Acute Toxicity: inhalation

Administrative data

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Justification for type of information:

Data is from study report.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Principles of method if other than guideline:

The study was conducted to find out the LC50, clinical effects and histopathological effect of test chemical at different aerosol concentration in Wistar albino rats.

GLP compliance:

no

Test type:

other: Acute Inhalation Toxicity

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Phenethyl phenylacetate

EC Number:

203-013-1

EC Name:

Phenethyl phenylacetate

Cas Number:

102-20-5

Molecular formula:

C16H16O2

IUPAC Name:

2-phenylethyl phenylacetate

Test material form:

liquid

Details on test material:

- Name of test material (as cited in study report): Phenethyl phenyl acetate
- Molecular formula : C16H16O2
- Molecular weight : 240.30 g/mole
- Substance type:Organic
- Physical state: Colorless Liquid
- Analytical purity: 99.55 %
- Lot/batch No.: 2012052101
- Storage condition of test material: Room temperature (20° - 30 °C)
- Other: Handling and Disposal
Safety precautions: Aprons, masks, caps, gloves and goggles were used to ensure the health and safety of the Personnel
Disposal:The remaining unused test item formulations were disposed as per internal SOPs

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: in bred in lab facility
- Age at study initiation: 7 to 9 weeks
- Weight at study initiation: Range of 200 ±20g
- Housing: Groups of 5 animals of similar sex in polypropylene cages with stainless steel grill top, facilities for food and water bottle, and bedding of clean paddy husk.
- Diet (e.g. ad libitum): conventional Laboratory diets, Pelleted feed supplied
- Water (e.g. ad libitum):Community tap water passed through ‘Aqua Guard on line water filter’, was kept in glass bottles, ad-libitum
- Acclimation period:20 healthy albino rats were selected and acclimatized for standard laboratory condition for period of 1 week in experimental room under veterinary examination.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-25 degree C
- Humidity (%): 30-60%
- Air changes (per hr): Air conditioned rooms with 10-15 air changes per hour
- Photoperiod (hrs dark / hrs light): illumination cycle set to 12 hours artificial fluorescent light and 12 hours dark.

Administration / exposure

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

nose only

Vehicle:

other: water (migrated information)

Remarks:

Distilled water

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: cylindrical chamber built from stainless steel and glass
- Exposure chamber volume: 8 liters
- Method of holding animals in test chamber: For the inhalation purpose the rats were placed in polycarbonate holder tubes positioned radically around exposure chamber, so that only the snouts and nostrils of the animals were exposed to the aerosol.
- Source and rate of air: The aerosol was generated by the used of Nanotek aerosol generator (particle size less than 1 micron)
- Method of conditioning air: The chamber was maintained at a slightly negative pressure to prevent leakage of the test atmosphere from the system, as well as its dilution with outside air.
- System of generating particulates/aerosols: Nanotek aerosol generator
- Treatment of exhaust air: The exhaust air was decontaminated by subsequent passage through 1% NaOH solution, silica gel and activated charcoal filters.

Analytical verification of test atmosphere concentrations:

not specified

Duration of exposure:

4 h

Concentrations:

Group I – Limit test (5 mg/L)
Group II – Confirmatory test (5 mg/L)

No. of animals per sex per dose:

10 (5 males and 5 females)

Control animals:

not specified

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical Signs: The treated animals were observed for signs of intoxication, at various intervals for first six hours after dosing and thereafter twice a day for 14 days. Any clinical signs in the treated group, if observed, recorded and mentioned in the report.
Body weight: The body weight of all the animals was observed weekly on day 0 (Before treatment), 7th and 14th (post treatment).
Mortality: All the animals were observed for mortality at various intervals for first six hours on the day of dosing and thereafter twice a day for 14 days.
- Necropsy of survivors performed: yes, Necropsy was carried out on all the animals that died during the study or surviving animals were sacrificed at the end of the study to observe any gross pathological changes.
- Other examinations performed: All the animals were observed for mortality and signs of intoxication at various intervals for first 6 hours on the day of dosing and thereafter twice a day for 14 days.

Statistics:

not specified

Results and discussion

Preliminary study:

not specified

Effect levels

Key result

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 5 mg/L air

Based on:

test mat.

Exp. duration:

4 h

Mortality:

No mortality at the tested dose level of 5.0 mg/L was observed throughout the period of observation after exposure.

Clinical signs:

other: other:

Body weight:

- other body weight observations: The body weight of all the animals recorded individually on day 7th and 14th (post treatment) showed normal gain as compared to day 0th.

Gross pathology:

No gross pathological changes were observed

Other findings:

NECROPSY FINDING
EXTERNAL
i. Skin-Skin and hair coat was observed wet.
ii. All external orifices-Normal
B. INTERNAL
i. Subcutaneous- No changes were observed.
ii. Superficial and deep lymph nodes- No change inmesenteric lymph node.
ABDOMINAL CAVITY
i. Opening and general examination- In the abdominal cavity all the organs were present in normal position.
ii. Spleen-No changes were recorded.
iii. Digestive system-No gross changes were observed in stomach and intestine.
iv. Liver and biliary ducts-No gross pathological changes were observed
v. Excretory system-No gross pathological changes were observed.
vi. Adrenal- Observed normal.
vii. Male/female genital organs –Showed normalcolour, consistency and no inflammatory changes.
2. THORACIC CAVITY
i. Opening and general examination- Thoracic cavity was found to be normal without any fluid, mucous or blood etc.
ii. Lungs-No changes were recorded.
iii. Heart- No changes were observed in color and consistency. Heart found normal.
iv. Thyroid- Normal in shape, size and surface.
3. CRANIAL CAVITY
i. Brain- Normal in shape and size.

Any other information on results incl. tables

TABLE -1

EXPOSURE ATMOSPHERE DATA

 

Parameters	Group-I (5 mg/L) (limit test)	Group-II (5 mg/L) (confirmatory test)
Chamber temperature °C (Mean ± S.E.)	23.19±1.46	24.21±1.63
Relative humidity (%)  (Mean ± S.E.)	46.27±2.35	45.27±2.69
Oxygen content (%) (Mean ± S.E.)	19.46±1.64	19.14±1.2

 

TABLE – 2

Mean Body Weight (gm)

 

S.No.	Groups	BODY WEIGHT (gm)
		DAY 0	DAY 7^th	% gain or loss	DAY14^th	% gain or loss
1.	Group-I (5.0 mg/L)	199.36	205.51	3.0 %	213.89	7.28 %
2.	Group-II (5.0 mg/L)	207.57	214.66	3.41 %	223.49	7.66 %

TABLE – 3

CLINICAL SIGNS AND MORTALITY

Group: I                                                                                           Dose: 5.00 mg/L

WISTAR ALBNINO RATS

 

Parameters	Incidence of Clinical Signs Observed after Dosing on																			Mortality
	Day 0					DAY
	Min	Hour
	30	1	2	4	6	1	2	3	4	5	6	7	8	9	10	11	12	13	14	Total	%
Mortality (total)	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0/10	0
Clinical Signs	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0

 

0         =   No clinical sign (Normal)

+         =   Mild

++        =   Moderate

+++      =   High

++++    =    Severe

Group: II                                                                                          Dose: 5.0 mg/L

                                                                                               

WISTAR ALBINO RATS

 

Parameters	Incidence of Clinical Signs Observed after Dosing on																			Mortality
	Day 0					DAY
	Min	Hour
	30	1	2	4	6	1	2	3	4	5	6	7	8	9	10	11	12	13	14	Total	%
Mortality (total)	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0/10	0
Clinical Signs	0	0	0	0	6	0	0	0	0	0	0	0	0	0	0	0	0	0	0

 

0         =   No clinical sign (Normal)

+         =   Mild

++        =   Moderate

+++      =   High

++++    =   Severe

TABLE – 4

SUMMARY OF NECROPSY FINDING

 

S. No.	Fate	Wistar albino rats
		Dose (mg/l)
		5.0 (limit test)	5.0 (confirmatory test)
1	Terminal sacrifice	10/10	10/10
2	Found Dead	0/10	0/10
3	Abnormalities detected	0/10	0/10

TABLE – 5

INDIVIDUAL ANIMAL FATE & NECROPSY FINDINGS

 

Group: I                                                                                                       5.0 mg/L

 

WISTAR ALBINO RATS

 

Animal ID	Fate	Time	Gross Findings
20167-1	TS	Day 14	NAD
20167-2	TS	Day 14	NAD
20167-3	TS	Day 14	NAD
20167-4	TS	Day 14	NAD
20167-5	TS	Day 14	NAD
20167-6	TS	Day 14	NAD
20167-7	TS	Day 14	NAD
20167-8	TS	Day 14	NAD
20167-9	TS	Day 14	NAD
20167-10	TS	Day 14	NAD

 

Day 0 is the day of exposure

TS=Terminal Sacrifice

NAD=No Abnormality Detected

FD=Fg/L

Group II- 5 mg/l 

WISTAR ALBINO RAT

Animal ID	Fate	Time	Gross Findings
20167-11	TS	Day 14	NAD
20167-12	TS	Day 14	NAD
20167-13	TS	Day 14	NAD
20167-14	TS	Day 14	NAD
20167-15	TS	Day 14	NAD
20167-16	TS	Day 14	NAD
20167-17	TS	Day 14	NAD
20167-18	TS	Day 14	NAD
20167-19	TS	Day 14	NAD
20167-20	TS	Day 14	NAD

 

Day 0 is the day of exposure

TS=Terminal Sacrifice

NAD=No Abnormality Detected

FD=Found Dead

Applicant's summary and conclusion

Interpretation of results:

other: Not classified

Conclusions:

The acute inhalation toxicity dose (LC50) was considered to be >5 mg/L (>5000 mg/m3), when male and female Wistar rats were exposed with the test chemical via inhalation route by nose only exposure for 4 hours. Hence the test chemical can be classified under the category “Not Classified” as per CLP Regulation.

Executive summary:

The acute inhalation study of test chemical was conducted in Wistar rats. The study was performed according to OECD-Guideline -403. In the limit test, ten healthy Wistar albino rats of both sexes (5 male and 5 female) of body weight 200±20 gm were selected for study after acclimatization. The test group was exposed to aerosol at the concentration of 5 mg/L for period of 4 hrs. After exposure all the animals were closely observed for clinical signs of toxicity at various intervals such as 1 hr, 2 hrs, 4 hrs, and 6 hrs on the day of test chemical aerosol exposure and later on twice a day throughout the experimentation period of 14 days.The necropsy was performed on all the animals which were died during the exposure or were sacrificed at termination of experiment. All the albino rats exposed to aerosol at the concentration of 5 mg/Ldid not show any clinical signs of intoxication. Furthermore, no mortality was observed throughout the period of observation. The necropsy was performed on all the animals at the termination of study did not show any gross pathological changes. After 72 hrs, the result obtained from limit test was confirmed in another 10 animal of both sex at similar concentration following same Guideline. Ten healthy Wistar albino rats of both sex body weight 200±20 gm were selected for study after acclimatization. The test groups of animals were exposed to aerosol at the concentration of 5 mg/L for period of 4 hrs. After exposure all the animals were closely observed for any clinical signs of toxicity at various intervals such as 1 hr, 2 hrs, 4 hrs, and 6 hrs on the day of test chemical aerosol exposure and later on twice a day throughout the experimentation period of 14 days.The necropsy was performed on all the animals at termination of experiment. All the albino rats exposed to aerosol at the concentration of 5mg/L did not show any clinical signs of intoxication. Furthermore, no mortality was observed throughout the period of observation.The body weight of animals exposed to test compound, observed on day 0th (pre treatment) and day 7th (post treatment) did not differ significantly as compared to day 0th. Whereas, body weight of animals observed on day 14th showed normal increase as compared to day 0th. The necropsy was performed on all the animals at the termination of study did not show any gross pathological changes.

The acute inhalation toxicity dose (LC50) was considered to be >5 mg/L (>5000 mg/m3), when male and female Wistar rats were exposed with the given test chemical via inhalation route by nose only exposure for 4 hours. From the results obtained from present investigation, it can be concluded that test chemical was non-toxic at the aerosol concentration of 5.0 mg/L under the test conditions.