Phenethyl acetate

Toxicological Summary

Workers - Hazard via inhalation route

Systemic effects

Long term exposure

Hazard assessment conclusion:

DNEL (Derived No Effect Level)

Value:

6.5 mg/m³

Most sensitive endpoint:

repeated dose toxicity

Route of original study:

Oral

DNEL related information

DNEL derivation method:

ECHA REACH Guidance

Overall assessment factor (AF):

75

Modified dose descriptor starting point:

LOAEC

Value:

490 mg/m³

Explanation for the modification of the dose descriptor starting point:

Repeated exposure via inhalation (or dermal application) route was not available.  Valid endpoints could be derived from orally administered studies (gavage or dietary) and for  different durations from 13 weeks to two years but the most relevant NOAEL was determined for female rats in a subchronic gavage study. While the NOAEL for males was 500 mg/kg bw/day the female rat NOAEL in the same study was 250 mg/kg bw/day and consequently this was used to derive the DNEL values by route-to route extrapolation based on the precautionary principle.  Since no local effects or short-term peak exposure effects were indicated, the long term DNEL is expected to be protective for short term exposure also.

AF for dose response relationship:

1

Justification:

NOAEL from subchronic study used as starting point

AF for differences in duration of exposure:

6

Justification:

extrapolation from subchronic to chronic exposure

AF for interspecies differences (allometric scaling):

1

Justification:

allometry not included where route-to-route extrapolation used to derive starting point

AF for other interspecies differences:

2.5

Justification:

default value

AF for intraspecies differences:

5

Justification:

default value

AF for the quality of the whole database:

1

Justification:

default value

AF for remaining uncertainties:

1

Justification:

default value

Acute/short term exposure

Hazard assessment conclusion:

no hazard identified

DNEL related information

Local effects

Long term exposure

Hazard assessment conclusion:

no hazard identified

Acute/short term exposure

Hazard assessment conclusion:

no hazard identified

DNEL related information

Workers - Hazard via dermal route

Systemic effects

Long term exposure

Hazard assessment conclusion:

DNEL (Derived No Effect Level)

Value:

2.27 mg/kg bw/day

Most sensitive endpoint:

repeated dose toxicity

Route of original study:

Oral

DNEL related information

DNEL derivation method:

ECHA REACH Guidance

Overall assessment factor (AF):

300

Modified dose descriptor starting point:

NOAEL

Value:

682 mg/kg bw/day

Explanation for the modification of the dose descriptor starting point:

Repeated exposure information via dermal application route was not available.  Valid endpoints could be derived from orally administered studies (gavage or dietary) and for  different durations from 13 weeks to two years but the most relevant NOAEL was determined for female rats in a subchronic gavage study. While the NOAEL for males was 500 mg/kg bw/day the female rat NOAEL was 250 mg/kg bw/day and consequently this was used to derive the DNEL values by route-to route extrapolation based on the precautionary principle.  Since no local effects or short-term peak exposure effects were indicated, the long term DNEL is expected to be protective for short term exposure also. The corrected starting dose based on relative absorption via oral and dermal routes was 682 mg/kg bw.

AF for dose response relationship:

1

Justification:

NOAEL from subacute study used as starting point

AF for differences in duration of exposure:

6

Justification:

extrapolation from subacute to chronic exposure

AF for interspecies differences (allometric scaling):

4

Justification:

default value for rat

AF for other interspecies differences:

2.5

Justification:

default value

AF for intraspecies differences:

5

Justification:

default value

AF for the quality of the whole database:

1

Justification:

default value

AF for remaining uncertainties:

1

Justification:

default value

Acute/short term exposure

Hazard assessment conclusion:

no hazard identified

DNEL related information

Local effects

Long term exposure

Hazard assessment conclusion:

no hazard identified

Acute/short term exposure

Hazard assessment conclusion:

no hazard identified

Workers - Hazard for the eyes

Local effects

Hazard assessment conclusion:

medium hazard (no threshold derived)

Additional information - workers

Toxicology summary

Toxicokinetics

Ester hydrolysis by artificial gastrointestinal juices followed first-order kinetics. The values for the ester hydrolysis rate constant (K) and t0.5 (time to effect 50% hydrolysis) showed a relatively slow rate of hydrolysis by artificial gastric juice and a faster reaction in artificial pancreatic juice. Phenylethyl acetate had a hydrolysis rate constant (K/h) of 0.139 and t0.5 of 300 mins in gastric juice and in pancreatic juice the values were 1.4 and 29.7 respectively. The results show relatively rapid hydrolysis of phenylethyl acetate by artificial gastro-intestinal juices and by liver and intestinal tissues and confirm the indications of rapid metabolic detoxification in the liver following repeat exposure and also the lack of bioaccummulation potential.

Oral absorption of benzyl acetate was approximately 90% based on urinary recovery of parent and metabolites. Dermal absorption was not quantified but was indicated to be considerably lower than oral absorption with marked losses due to volatilisation from appliaction sites and retention of unabsorbed dose on the dermal surface. An estimate of maximal dermal absorption was calculated and 33% dermal absorption is unlikely to be exceeded and may represent a combination of dermal penetration and unintended oral ingestion.

A default value of 100% absorption is assumed for the inhalation route.

Acute toxicity

Acute oral toxicity

Phenylethyl acetate LD50 was 5200 mg/kg in the rat.

On the basis of this study, phenylethyl acetate does not require classification according to Regulation (EC) No. 1272/2008.

Acute inhalation toxicity

No study data were available for phenylethyl acetate but an inhalation LC50 was available for the read-across candidate, benzyl acetate, that

indicated four hour exposure to phenyl acetate is unlikely to result in any systemic toxicity. The benzyl acetate exposure was 0.766 mg/L air, the maximum attainable concentration, which resulted in no adverse effects in rats.

Acute dermal toxicity

Dermal toxicity was assessed in groups of three rabbits dosed up to 15000 mg/kg bw. The acute dermal LD₅₀ in the rabbit was found to be 6210 mg/kg bw. Phenylethyl acetate

was found to be systemically toxic at doses higher than the regulatory limit dose but showed no indications of eliciting irritation or causing corrosive effects to dermal tissue. Classification for dermal toxicity is not required according to Regulation (EC) No. 1272/2008.

Irritation/Corrosion

In vivo administration of three doses to rabbits revealed 24 hour exposure to phenylethyl acetate at 6500, 10000 or 15000 mg/kg elicited no more than transient very slight erythema that resolved rapidly.

In another group of eight rabbits, treated with phenylethyl acetate for 4 hours under a semi-occlusive dressing, marginal or slight erythema and oedema were observed for up to 24 hours after dosing but reactions had reversed within 72 hours.

In an in vivo study of ocular irritation the reactions observed were not severe but did persist to termination and consequently phenylethyl acetate is considered to cause irreversible eye damage.

Phenylethyl acetate does not require classification as a skin irritant but should be classified for ocular effects as "Irreversible Effects on the Eye" Category 1 with corresponding hazard statement 'H319: Causes serious eye irritation', according to Regulation (EC) No. 1272/2008.

Sensitisation

In an open epicutaneous test the negative result confirmed the negative response in a human repeat insult test.

Phenylethyl acetate is not a skin sensitiser.

Repeated dose toxicity

Repeated dose oral toxicity

Data from studies with benzyl acetate were used to read-across to phenylethyl acetate.

Thirteen-week studies were conducted by Abdo (1986) to evaluate the cumulative toxicity of benzyl acetate and to determine the doses to be used in the 2-year studies. Four-week-old male and female F344/ N rats were obtained from Harlan Industries, observed for 13 days, and assigned by species and sex to cages according to a table of random numbers. The cages were then assigned to dosed and control groups according to another table of random numbers. Rats were housed five per cage in polycarbonate cages covered with spun-bonded polyester filters. Racks and filters were replaced every 2 weeks. Cages and bedding were replaced twice per week. Water, via an automatic watering system, and Wayne Lab Blox were available ad libitum. Groups of 10 rats of each sex were administered 0, 62.5, 125, 250, 500, or 1,000 mg/ kg benzyl acetate in corn oil by gavage, 5 days per week for 13 weeks. Animals were checked for mortality and signs of morbidity twice daily. Those animals that were judged moribund were killed and necropsied. Each animal was given a clinical examination weekly, including palpation for tissue masses or swelling. Individual body weight data were collected weekly. On days 92-96, survivors were killed with carbon dioxide. Necropsies were performed on animals that survived to the end of the study and on all animals found dead, unless precluded in whole or in part by autolysis or cannibalization. The following specimens were examined microscopically for controls, for the highest dosage group with at least 60% survivors at the time of the group kill, and for all animals that died before the survivors of the group were killed: gross lesions, tissue masses, abnormal lymph nodes, skin, mandibular lymph nodes, mammary gland, salivary gland, thigh muscle, bone, bone marrow, thymus, trachea, lungs and bronchi, heart,thyroid, parathyroid, esophagus, stomach, duodenum, jejunum, ileum, colon, mesenteric lymph nodes, liver, gallbladder (mice), pancreas, spleen, kidneys, adrenals, urinary bladder, seminal vesicles/ prostate/ testes or ovaries/ uterus, nasal cavity, brain, pituitary, and spinal cord, Tissues were preserved in 10% neutral buffered formalin, embedded in paraffin, sectioned, and stained with hematoxylin and eosin. The NOAEL was found to be 500 mg/kg body weight after dosing male rats with benzyl acetate in corn oil for 13 weeks.

The NOAEL was found to be 250 mg/kg body weight after dosing female rats with benzyl acetate in corn oil for 13 weeks. Based on these results, the test substance does not require classification according to Directive 67/548/EEC and Regulation EC No. 1272/2008.

The toxicity of benzyl acetate has also been investigated in high quality range-finding studies in the rat and mouse (NTP, 1993).

The NOAEL in female rats was 6250 ppm (equivalent to 480 mg/kg bw) based on reduced cholesterol levels in the 12500 ppm females. The NOAEL in male rats was 12500 ppm (equivalent to 900 mg/kg bw) based on reductions in body weight and food consumption in the 25000 ppm males.

The DNEL was derived from the gavage study conducted by Abdo in 1986, specifically from an NOAEL of 250 mg/kg bw/day (females considered more sensitive responders). Although other NOAEL values could be derived, for example from a carcinogenicity study, the subchronic exposure value in the oral gavage study was considered to represent the most sensitive endpoint in most sensitive sex, and therefore provide an appropriate starting point for the dose descriptor.

Repeated dose dermal toxicity

The lack of evidence of systemic toxicity or local effects in an acute dermal exposure study and the general lack of signs of dermal irritation in short term tests with several test species including rats, rabbits and guinea pigs combined with an absence of adverse occupational exposure reports of dermatitis or dermal irritation indicate that adverse effects are not anticipated in humans, either as dermal toxicity or irritancy, following repeated dermal exposure. No studies to investigate subacute or subchronic dermal systemic toxicity are presented and no dermal NOAEL was determined.

Repeated dose inhalation toxicity

Phenylethyl acetate has low volatility and lethal concentrations could not be generated for the acute exposure study. The maximum achievable concentration had no toxicologically significant effects. Physico-chemical limitations and animal welfare ethics preclude further assessment of repeated inhalation exposure.

On the basis of the repeated dose oral study results no classification is proposed for repeated dose toxicity.

Genetic toxicity

Phenylethyl acetate was investigated for potential to induce reverse mutations in bacterial tester strains in the Ames test. No positive results were obtained in direct plate incorporation or preincubation assays with or without the use of metabolic activation. the read across candidate material, benzyl acetate, was tested in the presence of metabolic activation in both the mouse and human lymphoma assays, and found to give a positive mutagenic response. However, in the absence of metabolic activation, the result was ambiguous since at higher dose levels, the benzyl acetate was toxic but was not considered to be mutagenic. Benzyl acetate also showed a marginal induction of structural aberrations, predominantly chromatid exchanges, but only at the highest dose in 24 hour continuous treatment without S9 mix. Because there was no structural aberration induction with any other treatment the

test substance was considered to be non-clastogenic. Since the in vitro results were inconclusive,

benzyl Acetate was investigated for its ability to induce unscheduled DNA synthesis in Fischer -344 rats following in vivo treatment.

Benzyl Acetate, was negative under the conditions employed in this study as the net grain/nucleus count was always negative and the percentage of cells undergoing repair was less than 10%.

On the basis of an overall negative response in a battery of genotoxicity tests, and using rad-across data where appropriate, phenylethyl acetate was not classified for genetic toxicity in accordance with Regulation (EC) No. 1272/2008.

Carcinogenicity

In a mouse pulmonary tumour incidence assay, groups of 20 mice were intraperitoneally injected with 24 doses of phenylethyl acetate in tricaprylin 2097. The dose levels were the MTD, 6000 mg/kg, and one-fifth MTD, 1200 mg/kg. Neither resulted in an increased incidence of pulmonary tumours in comparison with vehicle and untreated controls

There was no evidence of oncogenicity in the study, and therefore no classification is proposed for phrnylethyl acetate carcinogenicity.

Toxicity to reproduction

No specific studies of reproductive toxicity are available for phenylethyl acetate.

Effects on fertility

Various repeated administration studies have been conducted on the chemical and structural analogue, benzyl acetate.  In view of the chemical and structural similarities, it is considered that the available data are adequate for addressing phenylethyl acetate toxicity.

Specific investigations of reproductive function were made in both rats and mice at the end of the 13-week dietary studies. Sperm morphology and vaginal cytology of rats and mice from all dose levels were evaluated. No treatment-related effects on sperm morphology occurred in rats or mice. A significant dose-related effect occurred in oestrous cycling for female mice; this effect did not occur in female rats. The lengthening of the oestrous cycle observed in exposed female mice is considered to be related to the reduced body weights seen in this study and is not a direct toxic effect of the test material.

 

The data from the rat 13-week dietary study with benzyl acetate indicate a potential effect on the male reproductive tract. With the exception of a single male at 12500 ppm (with findings described as ‘minimal’), findings were associated with toxicity (including 90% mortality at the top dose level) and bodyweight effects and may therefore be secondary, rather than direct toxic effects of the test material. The single, minimal grade, incidence at 12500 ppm is not considered to be clearly related to treatment. It is notable that similar findings were not seen in the gavage study, which used comparable dose levels and in which the bolus dosing would be expected to produce more severe effects. The absence of effects in the rat following gavage or dietary administration for two years is equally notable, and specific investigations at the end of the 13-week dietary study did not reveal any effects on sperm morphology. No NOAEL values for reproductive toxicity were determined from these investigations.

Developmental toxicity

Benzyl acetate, was examined for its ability to induce teratogenic abnormalities when administered to female Wistar rats. The test substance was administered via oral gavage at dose concentrations of 0, 10, 100, 500 and 1000 mg/kg to groups of 20 -22 female rats on days 6 to 15 of gestation. No evidence of maternal toxicity was observed.

No evidence for developmental toxicity distinct from effects on dams although there

was a significant decrease observed in the weight of the males and females exposed to the top dose level.

.

Benzyl acetate, was not considered to be teratogenic under the conditions of this study. While there are indications that administration of 1000 mg/kg can influence the growth of the foetus, the effects were not significant. Based on these results, the test substance dose not require classification according to Regulation EC No. 1272/2008 or Directive 67/548/EEC.

DNEL derivation

The critical endpoint is considered to be the NOAEL of 250 mg/kg bw/d derived in the 13 week repeated dose oral toxicity study in rats. The value for females was used as it was lower than the corresponding male NOAEL of 500 mg/kg bw/day. 

Systemic effects observed included mortality and adverse clinical signs including trembling, sluggishness and ataxia; reduced bodyweights or weight gains and macroscopic pathology in the stomach although this was not supported by histopathological effects.

Local effects, resulting from repeated exposure to phenylethyl acetate or a structural analogue, were not apparent and there were no indicatiins of specific effects likely to arise from acute or peak exposures.

This study provides the lowest systemic NOAEL obtained in the available studies. Therefore, the oral NOAEL of 250 mg/kg bw/d is used as a starting point for derivation of the appropriate DNELs, including route-to-route extrapolation for inhalation and dermal exposure.The corrected starting point for inhalation was 490 mg/n3 and for dermal DNEL derivation was 682 mg/kg bw.

Oral absorption was set at a default value of 50% for phenyl ethyl acetate and dermal absorption was assumed to be at the same conservative level with a default assumption of 100% inhalation absorption also used in the DNEL calculations

according to REACH Guidance.

Correction for the relative extent of absorption is therefore not required when deriving systemic dermal DNEL values.

DNEL values were calculated using the oral NOAEL as the starting point with route-to-route extrapolation for other possible exposure scenarios. The long-term systemic DNELs derived using default assessment factors were also considered protective for short term exposure and against peak exposure exceedances.

Local effects did not require consideration.

General Population - Hazard via inhalation route

Systemic effects

Long term exposure

Hazard assessment conclusion:

DNEL (Derived No Effect Level)

Value:

1.61 mg/m³

Most sensitive endpoint:

repeated dose toxicity

Route of original study:

Oral

DNEL related information

DNEL derivation method:

ECHA REACH Guidance

Overall assessment factor (AF):

150

Modified dose descriptor starting point:

NOAEC

Value:

242 mg/m³

Explanation for the modification of the dose descriptor starting point:

Repeated exposure via inhalation (or dermal application) route was not available.  Valid endpoints could be derived from orally administered studies (gavage or dietary) and for  different durations from 13 weeks to two years but the most relevant NOAEL was determined for female rats in a subchronic gavage study. While the NOAEL for males was 500 mg./kg bw/day the female rat NOAEL was 250 mg/kg bw/day and consequently this was used to derive the DNEL values by route-to route extrapolation based on the precautionary principle.  Since no local effects or short-term peak exposure effects were indicated, the long term DNEL is expected to be protective for short term exposure also.

AF for dose response relationship:

1

Justification:

NOAEL used as starting point for DNEL derivation

AF for differences in duration of exposure:

6

Justification:

extrapolation from subacute to chronic exposure

AF for interspecies differences (allometric scaling):

1

Justification:

no adjustment for allometry required based on route-to-route extrapolation

AF for other interspecies differences:

2.5

Justification:

default value

AF for intraspecies differences:

10

Justification:

default value for general population

AF for the quality of the whole database:

1

Justification:

default value

AF for remaining uncertainties:

1

Justification:

default value

Acute/short term exposure

Hazard assessment conclusion:

no hazard identified

DNEL related information

Local effects

Long term exposure

Hazard assessment conclusion:

no hazard identified

Acute/short term exposure

Hazard assessment conclusion:

no hazard identified

DNEL related information

General Population - Hazard via dermal route

Systemic effects

Long term exposure

Hazard assessment conclusion:

DNEL (Derived No Effect Level)

Value:

1.14 mg/kg bw/day

Most sensitive endpoint:

repeated dose toxicity

Route of original study:

Oral

DNEL related information

DNEL derivation method:

ECHA REACH Guidance

Overall assessment factor (AF):

600

Modified dose descriptor starting point:

NOAEL

Value:

682 mg/kg bw/day

Explanation for the modification of the dose descriptor starting point:

Repeated exposure via inhalation (or dermal application) route was not available.  Valid endpoints could be derived from orally administered studies (gavage or dietary) and for  different durations from 13 weeks to two years but the most relevant NOAEL was determined for female rats in a subchronic gavage study. While the NOAEL for males was 500 mg./kg bw/day the female rat NOAEL was 250 mg/kg bw/day and consequently this was used to derive the DNEL values by route-to route extrapolation based on the precautionary principle.  Since no local effects or short-term peak exposure effects were indicated, the long term DNEL is expected to be protective for short term exposure also.

AF for dose response relationship:

1

Justification:

NOAEL used as starting point for DNEL derivation

AF for differences in duration of exposure:

6

Justification:

extrapolation from subacute to chronic exposure

AF for interspecies differences (allometric scaling):

4

Justification:

default value for rats

AF for other interspecies differences:

2.5

Justification:

default value

AF for intraspecies differences:

10

Justification:

default value

AF for the quality of the whole database:

1

Justification:

default value

AF for remaining uncertainties:

1

Justification:

default value

Acute/short term exposure

Hazard assessment conclusion:

no hazard identified

DNEL related information

Local effects

Long term exposure

Hazard assessment conclusion:

no hazard identified

Acute/short term exposure

Hazard assessment conclusion:

no hazard identified

General Population - Hazard via oral route

Systemic effects

Long term exposure

Hazard assessment conclusion:

DNEL (Derived No Effect Level)

Value:

0.42 mg/kg bw/day

Most sensitive endpoint:

repeated dose toxicity

Route of original study:

Oral

DNEL related information

DNEL derivation method:

ECHA REACH Guidance

Overall assessment factor (AF):

600

Modified dose descriptor starting point:

NOAEL

Value:

250 mg/kg bw/day

Explanation for the modification of the dose descriptor starting point:

Not required oral route used in key study for starting point derivation

AF for dose response relationship:

1

Justification:

NOAEL used as starting point for DNEL derivation

AF for differences in duration of exposure:

6

Justification:

extrapolation from subacute to chronic exposure

AF for interspecies differences (allometric scaling):

4

Justification:

default value for rats

AF for other interspecies differences:

2.5

Justification:

default value

AF for intraspecies differences:

10

Justification:

default value

AF for the quality of the whole database:

1

Justification:

default value

AF for remaining uncertainties:

1

Justification:

default value

Acute/short term exposure

Hazard assessment conclusion:

DNEL (Derived No Effect Level)

Value:

2.5 mg/kg bw/day

Most sensitive endpoint:

repeated dose toxicity

Route of original study:

Oral

DNEL related information

DNEL derivation method:

ECHA REACH Guidance

Overall assessment factor (AF):

100

Modified dose descriptor starting point:

NOAEL

Value:

250 mg/kg bw/day

Explanation for the modification of the dose descriptor starting point:

Not required oral route used in key study for starting point derivation

AF for dose response relationship:

1

Justification:

NOAEL used as starting point for DNEL derivation

AF for interspecies differences (allometric scaling):

4

Justification:

default value for rats

AF for other interspecies differences:

2.5

Justification:

default value

AF for intraspecies differences:

10

Justification:

default value

AF for the quality of the whole database:

1

Justification:

default value

AF for remaining uncertainties:

1

Justification:

default value

General Population - Hazard for the eyes

Local effects

Hazard assessment conclusion:

medium hazard (no threshold derived)

Additional information - General Population

Toxicology summary

Toxicokinetics

Ester hydrolysis by artificial gastrointestinal juices followed first-order kinetics. The values for the ester hydrolysis rate constant (K) and t0.5 (time to effect 50% hydrolysis) showed a relatively slow rate of hydrolysis by artificial gastric juice and a faster reaction in artificial pancreatic juice. Phenylethyl acetate had a hydrolysis rate constant (K/h) of 0.139 and t0.5 of 300 mins in gastric juice and in pancreatic juice the values were 1.4 and 29.7 respectively. The results show relatively rapid hydrolysis of phenylethyl acetate by artificial gastro-intestinal juices and by liver and intestinal tissues and confirm the indications of rapid metabolic detoxification in the liver following repeat exposure and also the lack of bioaccummulation potential.

Oral absorption of benzyl acetate was approximately 90% based on urinary recovery of parent and metabolites. Dermal absorption was not quantified but was indicated to be considerably lower than oral absorption with marked losses due to volatilisation from appliaction sites and retention of unabsorbed dose on the dermal surface. An estimate of maximal dermal absorption was calculated and 33% dermal absorption is unlikely to be exceeded and may represent a combination of dermal penetration and unintended oral ingestion.

A default value of 100% absorption is assumed for the inhalation route.

Acute toxicity

Acute oral toxicity

Phenylethyl acetate LD50 was 5200 mg/kg in the rat.

On the basis of this study, phenylethyl acetate does not require classification according to Regulation (EC) No. 1272/2008.

Acute inhalation toxicity

No study data were available for phenylethyl acetate but an inhalation LC50 was available for the read-across candidate, benzyl acetate, that

indicated four hour exposure to phenyl acetate is unlikely to result in any systemic toxicity. The benzyl acetate exposure was 0.766 mg/L air, the maximum attainable concentration, which resulted in no adverse effects in rats.

Acute dermal toxicity

Dermal toxicity was assessed in groups of three rabbits dosed up to 15000 mg/kg bw. The acute dermal LD₅₀in the rabbit was found to be 6210 mg/kg bw. Phenylethyl acetate

was found to be systemically toxic at doses higher than the regulatory limit dose but showed no indications of eliciting irritation or causing corrosive effects to dermal tissue. Classification for dermal toxicity is not required according to Regulation (EC) No. 1272/2008.

Irritation/Corrosion

In vivo administration of three doses to rabbits revealed 24 hour exposure to phenylethyl acetate at 6500, 10000 or 15000 mg/kg elicited no more than transient very slight erythema that resolved rapidly.

In another group of eight rabbits, treated with phenylethyl acetate for 4 hours under a semi-occlusive dressing, marginal or slight erythema and oedema were observed for up to 24 hours after dosing but reactions had reversed within 72 hours.

In an in vivo study of ocular irritation the reactions observed were not severe but did persist to termination and consequently phenylethyl acetate is considered to cause irreversible eye damage.

Phenylethyl acetate does not require classification as a skin irritant but should be classified for ocular effects as "Irreversible Effects on the Eye" Category 1 with corresponding hazard statement 'H319: Causes serious eye irritation', according to Regulation (EC) No. 1272/2008.

Sensitisation

In an open epicutaneous test the negative result confirmed the negative response in a human repeat insult test.

Phenylethyl acetate is not a skin sensitiser.

Repeated dose toxicity

Repeated dose oral toxicity

Data from studies with benzyl acetate were used to read-across to phenylethyl acetate.

Thirteen-week studies were conducted by Abdo (1986) to evaluate the cumulative toxicity of benzyl acetate and to determine the doses to be used in the 2-year studies. Four-week-old male and female F344/ N rats were obtained from Harlan Industries, observed for 13 days, and assigned by species and sex to cages according to a table of random numbers. The cages were then assigned to dosed and control groups according to another table of random numbers. Rats were housed five per cage in polycarbonate cages covered with spun-bonded polyester filters. Racks and filters were replaced every 2 weeks. Cages and bedding were replaced twice per week. Water, via an automatic watering system, and Wayne Lab Blox were available ad libitum. Groups of 10 rats of each sex were administered 0, 62.5, 125, 250, 500, or 1,000 mg/ kg benzyl acetate in corn oil by gavage, 5 days per week for 13 weeks. Animals were checked for mortality and signs of morbidity twice daily. Those animals that were judged moribund were killed and necropsied. Each animal was given a clinical examination weekly, including palpation for tissue masses or swelling. Individual body weight data were collected weekly. On days 92-96, survivors were killed with carbon dioxide. Necropsies were performed on animals that survived to the end of the study and on all animals found dead, unless precluded in whole or in part by autolysis or cannibalization. The following specimens were examined microscopically for controls, for the highest dosage group with at least 60% survivors at the time of the group kill, and for all animals that died before the survivors of the group were killed: gross lesions, tissue masses, abnormal lymph nodes, skin, mandibular lymph nodes, mammary gland, salivary gland, thigh muscle, bone, bone marrow, thymus, trachea, lungs and bronchi, heart,thyroid, parathyroid, esophagus, stomach, duodenum, jejunum, ileum, colon, mesenteric lymph nodes, liver, gallbladder (mice), pancreas, spleen, kidneys, adrenals, urinary bladder, seminal vesicles/ prostate/ testes or ovaries/ uterus, nasal cavity, brain, pituitary, and spinal cord, Tissues were preserved in 10% neutral buffered formalin, embedded in paraffin, sectioned, and stained with hematoxylin and eosin. The NOAEL was found to be 500 mg/kg body weight after dosing male rats with benzyl acetate in corn oil for 13 weeks.

The NOAEL was found to be 250 mg/kg body weight after dosing female rats with benzyl acetate in corn oil for 13 weeks. Based on these results, the test substance does not require classification according to Directive 67/548/EEC and Regulation EC No. 1272/2008.

The toxicity of benzyl acetate has also been investigated in high quality range-finding studies in the rat and mouse (NTP, 1993).

The NOAEL in female rats was 6250 ppm (equivalent to 480 mg/kg bw) based on reduced cholesterol levels in the 12500 ppm females. The NOAEL in male rats was 12500 ppm (equivalent to 900 mg/kg bw) based on reductions in body weight and food consumption in the 25000 ppm males.

The DNEL was derived from the gavage study conducted by Abdo in 1986, specifically from an NOAEL of 250 mg/kg bw/day (females considered more sensitive responders). Although other NOAEL values could be derived, for example from a carcinogenicity study, the subchronic exposure value in the oral gavage study was considered to represent the most sensitive endpoint in most sensitive sex, and therefore provide an appropriate starting point for the dose descriptor.

Repeated dose dermal toxicity

The lack of evidence of systemic toxicity or local effects in an acute dermal exposure study and the general lack of signs of dermal irritation in short term tests with several test species including rats, rabbits and guinea pigs combined with an absence of adverse occupational exposure reports of dermatitis or dermal irritation indicate that adverse effects are not anticipated in humans, either as dermal toxicity or irritancy, following repeated dermal exposure. No studies to investigate subacute or subchronic dermal systemic toxicity are presented and no dermal NOAEL was determined.

Repeated dose inhalation toxicity

Phenylethyl acetate has low volatility and lethal concentrations could not be generated for the acute exposure study. The maximum achievable concentration had no toxicologically significant effects. Physico-chemical limitations and animal welfare ethics preclude further assessment of repeated inhalation exposure.

On the basis of the repeated dose oral study results no classification is proposed for repeated dose toxicity.

Genetic toxicity

Phenylethyl acetate was investigated for potential to induce reverse mutations in bacterial tester strains in the Ames test. No positive results were obtained in direct plate incorporation or preincubation assays with or without the use of metabolic activation. the read across candidate material, benzyl acetate, was tested in the presence of metabolic activation in both the mouse and human lymphoma assays, and found to give a positive mutagenic response. However, in the absence of metabolic activation, the result was ambiguous since at higher dose levels, the benzyl acetate was toxic but was not considered to be mutagenic. Benzyl acetate also showed a marginal induction of structural aberrations, predominantly chromatid exchanges, but only at the highest dose in 24 hour continuous treatment without S9 mix. Because there was no structural aberration induction with any other treatment the

test substance was considered to be non-clastogenic. Since the in vitro results were inconclusive,

benzyl Acetate was investigated for its ability to induce unscheduled DNA synthesis in Fischer -344 rats followingin vivotreatment.

Benzyl Acetate, was negative under the conditions employed in this study as the net grain/nucleus count was always negative and the percentage of cells undergoing repair was less than 10%.

On the basis of an overall negative response in a battery of genotoxicity tests, and using rad-across data where appropriate, phenylethyl acetate was not classified for genetic toxicity in accordance with Regulation (EC) No. 1272/2008.

Carcinogenicity

In a mouse pulmonary tumour incidence assay, groups of 20 mice were intraperitoneally injected with 24 doses of phenylethyl acetate in tricaprylin 2097. The dose levels were the MTD, 6000 mg/kg, and one-fifth MTD, 1200 mg/kg. Neither resulted in an increased incidence of pulmonary tumours in comparison with vehicle and untreated controls

There was no evidence of oncogenicity in the study, and therefore no classification is proposed for phrnylethyl acetate carcinogenicity.

Toxicity to reproduction

No specific studies of reproductive toxicity are available for phenylethyl acetate.

Effects on fertility

Various repeated administration studies have been conducted on the chemical and structural analogue, benzyl acetate.  In view of the chemical and structural similarities, it is considered that the available data are adequate for addressing phenylethyl acetate toxicity.

Specific investigations of reproductive function were made in both rats and mice at the end of the 13-week dietary studies. Sperm morphology and vaginal cytology of rats and mice from all dose levels were evaluated. No treatment-related effects on sperm morphology occurred in rats or mice.Asignificant dose-related effect occurred in oestrous cycling for female mice; this effect did not occur in female rats. The lengthening of the oestrous cycle observed in exposed female mice is considered to be related to the reduced body weights seen in this study and is not a direct toxic effect of the test material.

 

The data from the rat 13-week dietary study with benzyl acetate indicate a potential effect on the male reproductive tract. With the exception of a single male at 12500 ppm (with findings described as ‘minimal’), findings were associated with toxicity (including 90% mortality at the top dose level) and bodyweight effects and may therefore be secondary, rather than direct toxic effects of the test material. The single, minimal grade, incidence at 12500 ppm is not considered to be clearly related to treatment. It is notable that similar findings were not seen in the gavage study, which used comparable dose levels and in which the bolus dosing would be expected to produce more severe effects. The absence of effects in the rat following gavage or dietary administration for two years is equally notable, and specific investigations at the end of the 13-week dietary study did not reveal any effects on sperm morphology. No NOAEL values for reproductive toxicity were determined from these investigations.

Developmental toxicity

Benzyl acetate, was examined for its ability to induce teratogenic abnormalities when administered to female Wistar rats. The test substance was administered via oral gavage at dose concentrations of 0, 10, 100, 500 and 1000 mg/kg to groups of 20 -22 female rats on days 6 to 15 of gestation. No evidence of maternal toxicity was observed.

No evidence for developmental toxicity distinct from effects on dams although there

was a significant decrease observed in the weight of the males and females exposed to the top dose level.

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Benzyl acetate, was not considered to be teratogenic under the conditions of this study. While there are indications that administration of 1000 mg/kg can influence the growth of the foetus, the effects were not significant. Based on these results, the test substance dose not require classification according to Regulation EC No. 1272/2008 or Directive 67/548/EEC.

DNEL derivation

The critical endpoint is considered to be the NOAEL of 250 mg/kg bw/d derived in the 13 week repeated dose oral toxicity study in rats. The value for females was used as it was lower than the corresponding male NOAEL of 500 mg/kg bw/day. 

Systemic effects observed included mortality and adverse clinical signs including trembling, sluggishness and ataxia; reduced bodyweights or weight gains and macroscopic pathology in the stomach although this was not supported by histopathological effects.

Local effects, resulting from repeated exposure to phenylethyl acetate or a structural analogue, were not apparent and there were no indicatiins of specific effects likely to arise from acute or peak exposures.

This study provides the lowest systemic NOAEL obtained in the available studies. Therefore, the oral NOAEL of 250 mg/kg bw/d is used as a starting point for derivation of the appropriate DNELs, including route-to-route extrapolation for inhalation and dermal exposure.

The corrected starting point for inhalation was 242 mg/n3 and for dermal DNEL derivation was 682 mg/kg bw.

Oral absorption was set at a default value of 50% for phenyl ethyl acetate and dermal absorption was assumed to be at the same conservative level with a default assumption of 100% inhalation absorption also used in the DNEL calculations

according to REACH Guidance.

Correction for the relative extent of absorption is therefore not required when deriving systemic dermal DNEL values.

DNEL values were calculated using the oral NOAEL as the starting point with route-to-route extrapolation for other possible exposure scenarios. The long-term systemic DNELs derived using default assessment factors were also considered protective for short term exposure and against peak exposure exceedances.

Local effects did not require consideration.