Reaction mass of m-cresol and 2-chloro-m-cresol and 6-chloro-m-cresol

Endpoint summary

Description of key information

In a reliable, GLP compliant OECD guideline 423  acute toxicity study with rats the oral LD50 of Preventol KMX was found to be greater than 300 mg/kg bw but below 2000 mg/kg bw.
In a reliable, GLP compliant OECD guideline 402  acute toxicity study with minor deviations the dermal LD50 of Preventol KMX was found to be greater than 2000 mg/kg bw for male rats and greater than 1000 mg/kg bw for female rats.
Therefore, Preventol KMX has to be classified according to Dangerous substance directive 67/548/EEC as harmful in contact with skin and if swallowed (Xn, R21/22) and according to CLP (1272/2008/EC) as harmful if swallowed (H302; acute oral Category 4) and harmful in contact with skin (H312; acute dermal Category 4). 
The test substance was not tested for acute toxicity via the inhalation route according to Regulation EC No 1907/2006 (REACH) Annex VIII, Section 8.5, Column 2.

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:

other: GLP compliant OECD 423 guideline study.

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

(2001)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

2009-02-09, North Rhine-westphalia

Test type:

acute toxic class method

Limit test:

no

Species:

rat

Strain:

other: Wistar HsdCpb:Wu

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan GmbH, 5960 AD Horts, Netherlands
- Age at study initiation: approx. 8 - 12 weeks (assumed by the body weight)
- Weight at study initiation: 156 g - 174 g
- Housing: grouped
- Diet: Provimi Kliba 3883 PM S15 Maus/Ratte Haltung, Kaiseraugust Switzerland; ad libitum, except for the day before administration of the test substance. Food was withheld from the animals for 16- 24 h before and 2-4 h after administration of the test substance.
- Water: tap water; ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2°C
- Humidity (%): 55 ± 5%
- Air changes (per hr): approx. 10
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

polyethylene glycol

Remarks:

PEG400

Details on oral exposure:

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

Doses:

300 and 2000 mg/kg bw

No. of animals per sex per dose:

3 females at 2000 mg/kg bw
2x3 females at 300 mg/kg bw

Control animals:

no

Details on study design:

- Starting dose: 2000 mg/kg bw
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical signs and mortality were determined several times at the day of application (defined as day 1) and at least once daily thereafter. The weight gain was checked weekly.
Animals which died or were sacrificed in a moribund state were weighed and subjected to gross pathology
- Necropsy of survivors performed: yes

Statistics:

A validated LAN-linked computer system was used for data collection, processing and evaluation.

Sex:

female

Dose descriptor:

LD50

Effect level:

> 300 - < 2 000 mg/kg bw

Based on:

test mat.

Mortality:

2/3 animals of the high dose group died during the observation period.
No mortalities occured at 300 mg/kg bw.

Clinical signs:

Clinical signs at 2000 mg/kg bw comprised of: decreased motility, lateral position, abdominal position, tremor, narrowed palpebral fissure, piloerection and labored breathing.
No clinical signs were observed in animals of the 300 mg/kg bw dose group.

Body weight:

No significant effects on body weight or body weight gain were noted.

Gross pathology:

Gross pathology of the two animals which died during the observation period revealed dilated intestine/stomach (gas filled). The animal of the high dose group sacrificed at study termination showed a spotted lung.
No gross pathological changes were noticed in any animal treated with 300 mg/kg bw.

Table 1: Table for acute oral toxicity.

Dose [mg/kg bw]	Toxicological results*	Duration of clinical signs	Time of death	Mortality (%)
Females
2000	2/3/3	10 min – 7 h	3 h – 6 h	67
300 (1^st )	0/0/3	--	--	0
300 (2^nd )	0/0/3	--	--	0
LD50 = >300 – 2000 mg/kg

* first number = number of dead animals                                 

 second number = number of animals with clinical signs         

 third number = number of animals used                               

Interpretation of results:

harmful

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

CLP: Acute oral Cat 4, H302
DSD: Xn, R22

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

300 mg/kg bw

Quality of whole database:

The LD50 was found to be >300 but below 2000 mg/kg bw. The database for this endpoint is deemed to be acceptable and sufficient for classification.

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Quality of whole database:

The test substance was not tested for acute toxicity via the inhalation route according to Regulation EC No 1907/2006 (REACH) Annex VIII, Section 8.5, Column 2.

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:

other: GLP compliant study which was stated to be according to OECD guideline 402. However, only males were tested at the limit dose of 2000 mg/kg bw while females were tested at 1000 mg/kg bw.

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Version / remarks:

(1987)

Deviations:

yes

Remarks:

In males the limit dose of 2000 mg/kg bw was tested. Females were tested with 1000 mg/kg bw. Either females should have been tested at the limit dose as well or a complete study design should have been performed to be in compliance with OECD 402.

GLP compliance:

yes (incl. QA statement)

Remarks:

2009-02-09, North Rhine-westphalia

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

other: Wistar HsdCpb:Wu

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan GmbH, 5960 AD Horts, Netherlands
- Age at study initiation: approx. 9 - 13 weeks (assumed by the body weight)
- Weight at study initiation: 290 g - 300 g (males), 203 - 213 g (females)
- Housing: individually
- Diet: Provimi Kliba 3883 PM S15 Maus/Ratte Haltung, Kaiseraugust Switzerland; ad libitum
- Water: tap water; ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2°C
- Humidity (%): 55 ± 5%
- Air changes (per hr): approx. 10
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: 30 cm^2 on the hair free back (approx. 10% of body surface)
- % coverage: 100%, (semiocclusive)

REMOVAL OF TEST SUBSTANCE
- Washing: The dressings were removed and the test site was rinsed with tepid water using soap and gently patting the area dry.
- Time after start of exposure: After 24 hours

TEST MATERIAL
- Amounts applied: The amount of applied pure test substance was calculated on basis of the actual body weight.
- Constant volume: No

Duration of exposure:

24 hours

Doses:

1000 mg/kg bw (female)
2000 mg/kg bw (males)

No. of animals per sex per dose:

5 males at 2000 mg/kg bw (dose range 19.3 - 20.0 mg/cm^2)
5 females at 1000 mg/kg bw (dose range 6.8 - 7.1 mg/cm^2)

Control animals:

not required

Details on study design:

- Duration of observation period following administration: 22 days
- Frequency of observations and weighing: Clinical signs and mortality were determined several times at the day of application (defined as day 1) and at least once daily thereafter. The weight gain was checked weekly.
Animals which died or were sacrificed in a moribund state were weighed and subjected to gross pathology
- Necropsy of survivors performed: yes

Statistics:

A validated LAN-linked computer system was used for data collection, processing and evaluation.

Sex:

male

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Sex:

female

Dose descriptor:

LD50

Effect level:

> 1 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: top dose used in females

Mortality:

1/5 males died during the observation period.
No death occured in females.

Clinical signs:

Clinical signs at 1000 mg/kg bw (females) and 2000 mg/kg (males) bw comprised of: decreased motility, bloody urine, narrowed palpebral fissure and piloerection.
Local signs at site of application at 2000 mg/kg bw (males) were: dark discoloration, thickening, a partial induration, swelling, encrustation and formation of scale skin.
Local signs at site of application at 1000 mg/kg bw (females) were: dark discoloration, thickening, a partial induration, encrustation and formation of scale skin.
For details see table 1 in section "Any other information on results"

Body weight:

The body weight and the body weight gain were decreased on day 8 of the study in males and females. For details see Table 2 in section "Any other information on results"

Gross pathology:

Gross pathology of the males which died during the observation period revealed dilated intestine (gas filled) and brownish-black discolored kidneys.The males and females sacrificed at study termination showed a partial formation of scale on the site of application.

Table 1: Incidence and duration of clinical signs

Parameter	Incidence	Duration of parameter
Males
decreased motility	5/5	2 h – 2 d
bloody urine	5/5	2 h – 7 h
narrowed palpebral fissure	5/5	3 h – 2 d
piloerection	1/5	2 d
Local effects
partial induration	4/5	2 d – 7 d
swelling	4/5	2 d
dark discoloration	3/5	2 d – 8 d
thickening	4/5	3 d – 7 d
scale skin	4/5	7 d – 22 d
encrustation	4/5	8 d – 21 d
Females
decreased motility	5/5	40 min – 2 d
bloody urine	5/5	40 min – 5 h
narrowed palpebral fissure	5/5	2 h – 5 h
piloerection	5/5	2 h – 2 d
Local effects
partial induration	5/5	3 d – 10 d
dark discoloration	5/5	2 d – 10 d
thickening	5/5	2 d
scale skin	5/5	11 d – 22 d
encrustation	5/5	3 d – 20 d

Table 2: Body weight and body weight gain

Day	1	8	15	22
Males
Mean body weight (g±SD)	296± 3.9	287± 6.5	327± 7.7	351± 6.2
Mean body weight gain (g±SD)	-	- 8± 6.7	41± 6.1	24± 2.3
Females
Mean body weight (g±SD)	207± 3.7	203± 3.6	215± 5.4	235± 9.2
Mean body weight gain (g±SD)	-	- 4± 1.2	12± 5.2	20± 10.6

SD = Standard deviation

Table 3: Table for acute dermal toxicity

Dose [mg/kg bw]	Toxicological results*	Duration of clinical signs	Time of death	Mortality (%)
Males
2000	1/5/5	2 h – 22 d	2d	20
Females
1000	0/5/5	40 min – 22 d	---	0
LD50 males >2000 mg/kg bw LD50 females >1000 mg/kg bw

* first number = number of dead animals                                 

second number = number of animals with clinical signs         

 third number = number of animals used     

Interpretation of results:

harmful

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

CLP: acute dermal Cat 4, H312
DSD: Xn, R21

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

1 000 mg/kg bw

Quality of whole database:

The LD50 was found to be >1000 mg/kg bw for females and >2000 mg/kg bw for males. The database for this endpoint is deemed to be acceptable and sufficient for classification.

Additional information

The relevant data on acute toxicity for Preventol KMX and the read across substance 4-chloro-3-methylphenol are summarized in the table below. The read across justification and additional data for m-cresol are attached to the Chemical Safety Report in Annex I. In comparison to the chlorocresols, m-cresol is only a minor component of Preventol KMX. Therefore these data are not considered for the assessment of the acute toxicity of Preventol KMX.

Table 1: Comparison of relevant data on acute toxicity of Preventol KMX and 4-chloro-3-methylphenol

Endpoint	Preventol KMX	4-chloro-3-methylphenol
Acute toxicity by oral route	LD50 rat: >300-2000 mg/kg bw (limit test)	LD50 rat: < 2000 mg/kg bw (limit test)
Acute toxicity by inhalation route	Data waiving due to corrosion	LD50 rat > 2871 mg/m³ (= maximum practically attainable aerosol concentration)
Acute toxicity by dermal route	LD50 rat: > 2000 mg/kg bw (male), > 1000 mg/kg bw (female)	LD50 rat: > 5000 mg/kg bw (male), > 2000 mg/kg bw (female)

 

Acute oral toxicity:

A reliable, GLP compliant acute oral toxicity study with doses of 300 and 2000 mg/kg bw Preventol KMX was performed according to OECD Guideline 423 with female young adult Wistar rats (Gillissen, 2011). In a first step 3 females received 2000 mg/kg bw Preventol KMX by oral gavage. Surviving animals were observed for clinical signs for 14 days, sacrificed thereafter and subjected to gross pathology. In a second step 2 groups of 3 females received 300 mg/kg bw test substance via oral gavage, were observed for clinical signs for 14 days, sacrificed thereafter and subjected to gross pathology.

After oral gavage of 2000 mg/kg bw Preventol KMX 2/3 females died within 3 - 6 hours after application. Clinical signs at 2000 mg/kg bw comprised of decreased motility, lateral position, abdominal position, tremor, narrowed palpebral fissure, piloerection and labored breathing. Gross pathology of the two animals which died revealed dilated intestine/stomach (gas filled). The animal of the high dose group sacrificed at study termination showed a spotted lung.

Upon oral gavage of 300 mg/kg bw no mortality and no clinical signs were observed throughout the observation period of 14 days. Also no gross pathological changes were noticed in females of this dose group.

Thus, the LD50 for rats was found to be greater than 300 mg/kg bw but below 2000 mg/kg bw.

Acute dermal toxicity:

A reliable, GLP compliant acute dermal toxicity study with doses of 1000 mg/kg bw Preventol KMX for 5 young adult female Wistar rats and 2000 mg/kg bw Preventol KMX for 5 young adult male Wistar rats was performed according to OECD Guideline 402 (Gillissen, 2011) with minor deviations.

Animals of both dose groups were exposed to the pure test substance under semi occlusive conditions at the hair free back and the test substance was removed by washing with tepid water and soap after 24 hours. Animals were observed for mortality and clinical signs of toxicology for 22 days. Body weights were assessed once weekly. All animals were sacrificed after the observation period and were, as the animals found dead during the observation period, subjected to gross pathology.

1/5 males died within 2 hours after application while no female died within the study period. Clinical signs for males and females comprised of decreased motility, bloody urine, narrowed palpebral fissure and piloerection. Local signs at site of application were dark discoloration, thickening, a partial induration, swelling (only males), encrustation and formation of scale skin. The body weight and the body weight gain were decreased on day 8 of the study in males and females. Gross pathology of the male which died during the observation period revealed dilated intestine (gas filled) and brownish-black discolored kidneys. The males and females sacrificed at study termination showed a partial formation of scale on the site of application.

Thus, the LD50 for female rats was found to be greater than 1000 mg/kg bw and the LD50 for males is greater than 2000 mg/kg bw. As there is no possibility to assess whether or not the LD50 for females is, as for the males, above 2000 mg/kg bw Preventol KMX has to be classified as harmful to the skin.

Acute inhalation toxicity:

According to Regulation EC No 1907/2006 (REACH) Annex VIII, Section 8.5, Column 2 testing for acute toxicity does generally not need to be conducted if “the substance is classified as corrosive to the skin”.

Justification for selection of acute toxicity – oral endpoint
Reliable GLP compliant OECD Guideline 423 key study.

Justification for selection of acute toxicity – dermal endpoint
Reliable GLP compliant OECD Guideline 402 key study.

Justification for classification or non-classification

Preventol KMX has to be classified according to Dangerous substance directive 67/548/EEC as harmful in contact with skin and if swallowed (Xn, R21/22) and according to CLP (1272/2008/EC) as harmful if swallowed (H302; acute oral Category 4) and harmful in contact with skin (H312; acute dermal Category 4).