Oligomerisation and alkylation reaction products of 2-phenylpropene and phenol

Endpoint summary

Description of key information

The substance oligomerisation and alkylation reaction products of 2-phenylpropene and phenol (OAPP) (previous name phenol, methylstyrenated) shows low oral, inhalation, and dermal acute toxicity. Discriminating doses (oral studies) were determined to be 2000 mg/kg bw (no mortality observed). Application of the substance as aerosol at a concentration of 4.92 mg/L did not result in any mortality (LC50 > 5000 mg/m³). The LD50 obtained in an acute dermal toxicity study was > 2000 mg/kg bw. Combined results indicate that OAPP is of low acute toxicity independent of the application route.

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:

22 Jul. - 14 Aug. 2007

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

Limit test:

yes

Specific details on test material used for the study:

- Name of test material (as cited in study report): Novares LA 300 (phenol, methylstyrenated)
- Lot/batch No.: 24087
- Composition of test material: composition is specified in IUCLID Sect. 13 - Assessment reports under Certificate of Analysis_Novares LA 300_phenol, methylstyrenated
- Stability under test conditions: no measured data; based on chemical structure assumed to be stable
- Storage condition of test material: room temperature, exclusion of light

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Winkelmann GmbH, Borchen/Germany
- Age at study initiation: young adult
- Weight at study initiation: 161 - 205 g
- Fasting period before study: overnight
- Housing: cages
- Diet: ad libitum, except 3 - 4 h after treatment
- Water: ad libitum
- Acclimation period: no data


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +-3
- Humidity (%): 55 +-10
- Air changes (per hr): 10x
- Photoperiod (hrs dark / hrs light): 12 / 12

Route of administration:

oral: gavage

Vehicle:

cotton seed oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 2 g/10 mL
- Amount of vehicle (if gavage): ~8 mL/kg bw
- Justification for choice of vehicle: miscible with the TS
- Lot/batch no. (if required): 066K0057 (Sigma Chemicals Co.)

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

6 (female)

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: 0, 7, and 14 d
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight

Statistics:

not applicable

Key result

Sex:

female

Dose descriptor:

discriminating dose

Effect level:

2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: Novares LA 300

Mortality:

none

Clinical signs:

other: Diarrhoea in 1/6 animals after 1 - 2 days post-application

Gross pathology:

no particular findings

Other findings:

none

Interpretation of results:

Category 5 based on GHS criteria

Remarks:

Migrated information

Conclusions:

no classification required

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

2 000 mg/kg bw

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)

Qualifier:

according to guideline

Guideline:

EU Method B.2 (Acute Toxicity (Inhalation))

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

yes

Specific details on test material used for the study:

- Name of test material (as cited in study report): Novares LA 300 (phenol, methylstyrenated)
- Lot/batch No.: 28166
- Composition of test material: composition is specified in IUCLID Sect. 13 - Assessment reports under Certificate of Analysis_Novares LA 300_phenol, methylstyrenated
- Stability under test conditions: no measured data; based on chemical structure assumed to be stable
- Storage condition of test material: room temperature, exclusion of light

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Laboratories Ltd, Oxon, UK.
- Age at study initiation: 8 - 12 wks
- Weight at study initiation: 270 - 284 g (m); 199 - 220 (f) [see Appendix 6]
- Fasting period before study: no
- Housing: solid-floor polypropylene cages, groups of 5 by sex
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: >= 5 d


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 25
- Humidity (%): 30 - 70
- Air changes (per hr): >= 15x/h
- Photoperiod (hrs dark / hrs light): 12 / 12

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

nose only

Vehicle:

other: diethyl ether

Mass median aerodynamic diameter (MMAD):

2.22 µm

Geometric standard deviation (GSD):

2.83

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: Cylindrical exposure chamber
- Exposure chamber volume: approx. 30 litres (dimensions: 28 cm diameter x 50 cm high)
- Method of holding animals in test chamber: individually held in a tapered, polycarbonate restraining tube

- Method of conditioning air: Compressed air supplied by means of an oil free compressor and passed
through a water trap and respiratory quality filters before it was introduced to the nebuliser.
- Air flow through chamber: 60 L/min

- System of generating particulates/aerosols: Glass concentric jet nebuliser located at the top of the exposure chamber /
Nebuliser connected to a glass syringe attached to an infusion pump
providing the material formulation

- Method of particle size determination: 3x during the exposure, using a Marple Personal Cascade Impactor
with six impactor stages (9.0, 6.3, 4.0, 1.7, 0.81 and 0.30 µm cut points).
The collection substrates and backup filter were weighed before and after sampling
and the weight of test material, collected at each stage, calculated by difference
(Results in Report Fig. 3 and 4).

- Method of particle collection: by weighed glass fibre filter placed in a filter holder and temporarily sealed
in a vacant port of the exposure chamber in the animals’ breathing zone.

- Volatile / non-volatile fraction: The mean non-volatile component of the batch used during the study was found to be 98.6 % (n=10).

Procedure: Prior to the start of the study, the non-volatile component of the test material was determined
by adding a small, known amount of test material to glass fibre filters and recording their weights.
The filters were then kept in a desiccator between 19 and 20°C for approx. 24 h and then weighed again.
The difference in the two weights was taken as the volatile content of the test material and the non-volatile component
was calculated as a percentage.

- Treatment of exhaust air: bottom outlet through a "scrubber" trap, connected with a high efficiency filter to a metered exhaust system
- Temperature, humidity, pressure in air chamber: 19 - 20 °C, 39 - 47% (rel.hum.), slight low-pressure [Report, Appendix 9]


TEST ATMOSPHERE
- Brief description of analytical method used: particle/aerosol gravimetric determination
- Samples taken from breathing zone: yes


VEHICLE
- Composition of vehicle (if applicable): diethyl ether
- Concentration of test material in vehicle (if applicable): 50 % (w/w)
- Justification of choice of vehicle: high viscosity of the TS, to improve aerolisation of the TS


TEST ATMOSPHERE (if not tabulated)
- Particle size distribution: ca. 72% (w/w) with aerodynamic diameter < 4 µm
- MMAD (Mass median aerodynamic diameter) / GSD (Geometric st. dev.): 2.22 / 2.38 µm

Analytical verification of test atmosphere concentrations:

yes

Remarks:

gravimetrically (aerosol)

Duration of exposure:

4 h

Concentrations:

Mean: 4.92 +-0.46 mg/L (n = 17) / nominal: 50.3 mg/L

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: day 0, 7, 14
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other:

Statistics:

For particle size: arithmetic mean + standard deviation / MMAD derived from probits of stage amounts plotted against Log10 cut-point size + geometric standard deviation / LD50: not relevant

Key result

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 4.92 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Remarks on result:

other: Novares LA 300; standard deviation of analytical determination: 0.46 mg/L

Sex:

male/female

Dose descriptor:

LC0

Effect level:

4.92 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Remarks on result:

other: Novares LA 300; standard deviation of analytical determination: 0.46 mg/L

Mortality:

none

Clinical signs:

other: Hunched posture, pilo-erection, and increased respiratory rate commonly seen for short periods following 4h inhalation; isolated instance of lethargy / All female animals exhibited staining of the head and one also showed noisy respiration. 4 - 6 d after

Body weight:

One male with bw transiently reduced during week 1.
Normal bodyweight development was noted for all other animals during the course of the study.
[see Report Appendix 6]

Gross pathology:

No macroscopic abnormalities

The particle size analysis of the atmosphere drawn from the animals’ breathing zone, was as follows

[see Report Appendices 1 and 2]:

Mean Achieved Atmosphere Concentration +-SD [mg/L] (n = 17)	Mean Mass Median Aerodynamic Diameter [µm] (n = 3)	Inhalable Fraction [wt% <4 µm]	Geometric Standard Deviation [µm]
4.92 +-0.46	2.22	71.5	2.83

Interpretation of results:

Category 5 based on GHS criteria

Conclusions:

No classification required

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LC50

Value:

5 000 mg/m³ air

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:

19. 10. – 05. 11. 2009

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

yes

Specific details on test material used for the study:

- Name of test material (as cited in study report): Novares LA 300 (phenol, methylstyrenated)
- Lot/batch No.: 28166
- Composition of test material: composition is specified in IUCLID Sect. 13 - Assessment reports under Certificate of Analysis_Novares LA 300_phenol, methylstyrenated- Stability under test conditions: no measured data; based on chemical structure assumed to be stable
- Storage condition of test material: room temperature, exclusion of light

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Breeding farm VELAZ s.r.o., Koleč u Kladna, Czech Republic
- Age at study initiation: no data, adult
- Weight at study initiation: 278 - 317 g (m); 203 - 223 g (f);
- Fasting period before study: no
- Housing: 1 animal/plastic cage
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: at least 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +-3
- Humidity (%): 30 - 70
- Photoperiod (hrs dark / hrs light): 12 / 12

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: about 6 x 6 cm
- % coverage: aprox. 10% of the body surface
- Type of wrap if used: mull and plaster (strapping)


REMOVAL OF TEST SUBSTANCE
- Washing (if done): water
- Time after start of exposure: 24 h

Duration of exposure:

24 h

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Body weight: before application, 8th and 15th day of study
Mortality: daily
Clinical signs: daily
Pathological examination: 15th day of study

- Necropsy of survivors performed: yes

Statistics:

not applicable

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: Novares LA 300

Sex:

male/female

Dose descriptor:

LD0

Effect level:

2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: Novares LA 300

Mortality:

none

Clinical signs:

other: after 3 h: piloerection in 9/10 animals, single cases with red secretion around eyes and decreased response to stimuli; after 2 d: no particular findings, but 1 female with skin irritation which healed by day 11.

Gross pathology:

no particular findings, but 1 female with discoloration of the liver (light colour)

Table No. 1: Individual body weight of animals – 2000 mg/kg – males

Animal No.	Before application	8th day	15th day	Body weight gain (g)
				day 0-8 p.a.	day 8-15 p.a.
1 (pre-test)	298.75	317.19	340.74	18.44	23.55
2	278.31	298.94	313.65	20.63	14.71
3	304.74	317.66	345.98	12.92	28.32
4	316.89	332.24	354.09	15.35	21.85
5	303.66	311.04	330.31	7.38	19.27
Average	300.47	315.41	336.95	14.94	21.54

Table No. 2: Individual body weight of animals – 2000 mg/kg – females

Animal No.	Before application	8th day	15th day	Body weight gain (g)
				day 0-8 p.a.	day 8-15 p.a.
1 (pre-test)	223.08	219.18	231.28	-3.90	12.1
2	223.42	216.23	223.05	-7.19	6.82
3	217.08	226.69	232.16	9.61	5.47
4	203.13	200.73	214.09	-2.40	13.36
5	204.65	200.79	204.00	-3.86	3.21
Average	214.27	212.72	220.92	-1.55	8.19

The test substance applied on skin at a dose of 2000 mg/kg of animal weight did not cause death of animals.    

Clinical signs of intoxication (piloerection, decreased response to stimuli, red secretion around eyes) were observed in all males and four females. Irritation on the skin was observed after application of the test substance in one female. Symptoms of irritation faded away on 12^thday after application of the test substance. Decreased body weight in females was recorded in period day 0-8 of the study. Macroscopic changes were diagnosed during pathological examination in one female (liver – light colour).

Interpretation of results:

Category 5 based on GHS criteria

Conclusions:

no classification required

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

2 000 mg/kg bw

Additional information

Acute Oral

Four GLP compliant studies have been conducted on the acute oral toxicity endpoint using the substance oligomerisation and alkylation reaction products of 2-phenylpropene and phenol (previous name phenol, methylstyrenated) (specifically; three OECD 423 and one OECD 401). In one study, 2 animals died following treatment (though no specific cause of death was noted). No other deaths occurred in the three other studies. The discriminating doses/LD50 recorded for each study are 2000 and greater than 2000 mg/kg bw, respectively. The overwhelming weight of evidence would suggest that OAPP is not likely to be acutely toxic via the oral route.

Acute Dermal

Two GLP compliant studies (OECD 402) have been conducted on the acute dermal toxicity endpoint using the substance oligomerisation and alkylation reaction products of 2-phenylpropene and phenol. No deaths were recorded in either study. As such it can be concluded that OAPP has a low acute dermal toxic potential. The LD50 in both studies was greater than 2000 mg/kg bw, and no significant treatment related toxicities were seen.

Acute Inhalation

A GLP compliant study was conducted on the substance oligomerisation and alkylation reaction products of 2-phenylpropene and phenol (as an aerosol) via the inhalation route. No animal deaths were noted during the study. The maximum dose given was 4.92 mg/L per animal +/- 0.46 mg/L. The LD50 was estimated to be greater than 4.92 mg/L. For classification purposes, it has been assumed that the LD50 is greater than 5.00 mg/L. This is because there was no mortality and the margins of error quoted for the study exceed the threshold for non-classification.

Justification for classification or non-classification

For all three application routes, data from acute toxicity limit tests with concentrations set to the upper limit for classification are available. Mortality in all tests was absent or low. For oral application, the discriminating dose in three studies was 2000 mg/kg bw. For inhalation and dermal route, distinct LD(C)50 values could not be determined (LD(C)50 > limit concentration). Based on these results, classification according to Regulation (EC) 1272/2008 (CLP Regulation) is not required.