Formaldehyde, oligomeric reaction products with phenol

Endpoint summary

Description of key information

Skin Irritation:

The dermal irritation potential of test article was determined according to the OECD 439 test guideline for this study. The MatTek EpiDerm™ model was used to assess the potential dermal irritation of the test article by determining the viability of the tissues following exposure to the test article via MTT. Tissues were exposed to the test article and controls for ~one hour, followed by a 42 hour post-exposure recovery period. The viability of each tissue was determined by MTT assay. The MTT data show the assay quality controls were met and passed the acceptance of criteria. The Mean % tissue viability compared to negative control (n=3) of the test substance Phenol-formaldehyde resin (CAS No. 9003 -35 -4) was determined to be 103.9 %. Hence, under the current experimental test conditions it was concluded that test substance Phenol-formaldehyde resin (CAS No. 9003 -35 -4) was considered to be not irritating to human skin and thus “Not Classified'' as per CLP Regulation.

Eye Irritation:

The treated animals produced slight ocular reactions with the average score was 10.6 out of maximum score of 110. Hence, chemical was considered to be Slightly irritating to the albino rabbits.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Data is from experimental study report

Qualifier:

according to guideline

Guideline:

OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)

Principles of method if other than guideline:

The purpose of this study was to assess potential for the test article to be dermal irritants. The dermal irritation potential of test article may be predicted by measurement of their cytotoxic effect, as reflected in the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, in the MatTek EpiDerm™ model.

GLP compliance:

no

Specific details on test material used for the study:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature or Fridge storage

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: Prior to the main test, the test articles are tested for their ability to reduce/interact with MTT and their ability to stain the tissues itself. All tests are performed according to the by MatTek provided test protocol.

FORM AS APPLIED IN THE TEST: Solid


- Name of test material (as cited in study report): Phenol-formaldehyde resin
- Molecular formula: C6H6O.CH2O)x-
- Molecular weight: 34.133 g/mol
- Substance type: organic
- Physical state: solid
- Smiles notation: c1(c(c(ccc1)[CH])O)O[CH]
- InChl: 1S/C8H6O2/c1-6-4-3-5-7(10-2)8(6)9/h1-5,9H

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: as provided by MatTek In Vitro Life Science Laboratories, Bratislava, Slovakia

Source strain:

other: Not applicable

Details on animal used as source of test system:

- Description of the cell system used: The normal human-derived keratinocytes were cultured at the air-liquid interface in a chemically defined medium on a permeable polycarbonate insert (surface 0.5 cm2). They were cultured in chemically defined serum free medium to form a multi-layered epithelium similar to that found in native epidermis. Each lot of tissues was Quality Assured by MatTek according to specific QC standards including: histology, tissue viability (MTT mean optical density), reproducibility (SD) and tissue thickness.Test System IdentificationAll of the EpiDerm™ 3-dimensional human tissues used in this study were identified by the date of arrival and the lot number. Certificate of Analysis for the tissues are included in this report. Tissue plates were appropriately labeled with study information.

Justification for test system used:

The 3-Dimensional Human Dermal Epithelial Model (EpiDerm™, MatTek, In Vitro Life Science Laboratories, Bratislava, Slovakia) is made up of normal human keratinocytes in serum free medium. The cells form an epithelial tissue that consists of organized basal, spinous, granular, and cornified layers analogous to those found in vivo. The EpiDerm™ model also contains epidermis-specific differentiation markers such as pro-filaggrin, the K1/K10 cytokeratin pair, involucrin, and type I epidermal transglutaminase, as well as keratohyalin granules, tonofilament bundles, desmosomes, and a multi-layered stratum corneum containing intercellular lamellar lipid layers arranged in patterns characteristic of in vivo epidermis. Each lot of tissues was Quality Assured by MatTek, Inc. according to specific QC standards including: histology (cell layers), tissue viability (MTT mean optical density) and reproducibility (SD). Tissue plates were appropriately labeled with study information. Bias was not a factor in this test system.

Vehicle:

unchanged (no vehicle)

Details on test system:

The tissues were exposed to the test article neat (undiluted) on April 25, 2018 (Run 1 of 1). EpiDerm™ tissues were purchased from MatTek. Quality control of the tissues was performed by MatTek and the Certificate of Analysis (CoA) for the tissues is provided and is kept in the study binder. Tissues were exposed for approximately 1 hour, with 35 minutes in an approximately 37°C, 5% CO2 humidified incubator and the remaining 25 minutes at room temperature. Following the exposure time, the tissues were rinsed and placed in fresh media for approximately 24 hours. The media was then changed again and the tissues were incubated in fresh media for another ~18 hours for a total of approximately 42 hour post-exposure recovery period. The tissue viability was then assessed by MTT assay. The tissue CoA was used instead of verification of barrier properties of the tissue.MTT and Color Pre-tests Pretesting has actually been conducted for all chemicals, although the first intitial 8 test chemicals a pretesting was not conducted (for skin). MTT AssayFollowing the rinsing period, the MTT assay was performed by transferring the tissues to 24-well plates containing 300 µL MTT medium (1.0 mg/mL). After 3 hours MTT incubation at approximately 37°C, approximately 5% CO2 in a humidified incubator, the blue formazan salt was extracted by submerging tissues in 2 mL isopropanol in a 24-well plate. The extraction time was approximately 3 hours with gentle shaking. The optical density of the extracted formazan (200 µL/well of a 96-well plate) was determined using a Thermo Scientific Multiskan FC Microplate Photometer at 570 nm. Relative cell viability is calculated for each tissue as % of the mean negative control tissues.Evaluation of Test Article in the Cell Models:

1. Cell system: Upon receipt, the MatTek EpiDerm™ tissue cultures were placed in 0.9 mL of fresh Maintenance medium (in a 6-well plate). The culture inserts are incubated for ~one hour. The tissues were then transferred to 6-well plates containing 0.9 mL fresh Maintenance medium and they were incubated overnight (18 ± 3 hrs) at ~37°C, 5% CO2 in a humidified incubator.

2. Control and Test Article Exposures: On the day of dosing, the tissues are then removed from the incubator and the controls and the test article are applied topically to tissues by pipette(liquid) Tissues were exposed to controls and the test article for one hour, with ~35 minutes in a 37°C, 5% CO2 humidified incubator and the remaining 25 minutes at room temperature.a) Controls 30 µL of negative control DPBS and 30 μL of the positive control 5% SDS was applied topically to the tissue and gently spread by placing a nylon mesh on the apical surface of each tissue, if necessary.b) Test Articles 25 mg of the test article was applied topically to the tissue

3. Post-exposure treatmentAfter the 1 hour exposure, the tissues were rinsed 15 times with sterile DPBS. After the 15th rinse from washing bottle, each insert wasw completely submerge 3 times in 150 ml DPBS. The apical surface was gently blotted with a cotton swab. The tissues were placed in 0.9 mL of fresh Maintenance medium (6-well plate) for 24 ± 2 hours. After this initial ~24 hour incubation, the tissues were placed in 6-well plates containing 0.9 mL fresh Maintenance medium and incubated for another 18 ± 3 hours, for a total of an approximately 42 hour post-exposure incubation.

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: The EpiDerm™ 3 dimensional human tissue model
- Tissue Lot number(s): 26459
- Date of initiation of testing: 6/08/2017

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37°C
- Temperature of post-treatment incubation (if applicable): 37°C

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: The test substance was rinsed from the tissues with sterile DPBS by filling and emptying the tissue insert 15 times to remove any residual test material. This was followed by completely submerge the insert 3 times in 150 ml DPBS.

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE

- MTT concentration: 300 µL MTT medium (1.0 mg/mL)
- Incubation time: After 3 hours
- Spectrophotometer: Synergy H4 spectrophotometer
- Wavelength: 570 nm
- Filter: No data
- Filter bandwidth: No data
- Linear OD range of spectrophotometer: No data

NUMBER OF REPLICATE TISSUES: 3
CALCULATIONS and STATISTICAL METHODS
All data were background subtracted before analysis. MTT data are presented as % viable compared to negative control. Data were generated as follows: MTT AssayBlanks:·        The optical density (OD) mean from all replicates for each plate (ODblank).
Negative Controls (NC):·        The blank corrected value was calculated: ODNC= ODNCraw– ODblank. ·        The OD mean per NC tissue was calculated. ·        The mean OD for all tissues corresponds to 100% viability. ·        The mean, standard deviation (SD), standard error of the mean (SEM) and the percent coefficient of variation (% CV) was calculated. Positive Control (PC):·        Calculate the blank corrected value: ODPC= ODPCraw– ODblank. ·        The OD mean per PC tissue was calculated. ·        The viability per tissue was calculated: %PC = [ODPC/ mean ODNC] x 100. ·        The mean viability for all tissues was calculated: Mean PC = Σ %PC / number of tissues. ·        The standard deviation (SD), standard error of the mean (SEM) and the percent coefficient of variation (% CV) was calculated. Tested compound :·        Calculate the blank corrected value ODTT= ODTTraw– ODblank. ·        The OD mean per tissue was calculated. ·        The viability per tissue was calculated: %TT = [ODTT/ mean ODNC] x 100. ·        The mean viability for all tissues was calculated: Mean TT = Σ %TT / number of tissues. ·        The standard deviation (SD), standard error of the mean (SEM) and the percent coefficient of variation (% CV) was calculated. Data Correction Procedure for MTT Interfering Compounds (if applicable)True viability = Viability of treated tissue – Interference from test article = ODtvt– ODktwhere ODkt= (mean ODtkt– mean ODukt).ODtvt= optical density of treated viable tissueODkt= optical density of killed tissuesODtkt= optical density of treated killed tissueODukt= optical density of untreated killed tissue (NC treated tissue) Data Correction Procedure for Colored Compounds (if applicable)True viability = Viability of treated tissue incubated in MTT media – Viability of treated tissue incubated in media without MTT = ODtvt– ODvt.ODtvt= optical density of treated viable tissue incubated in MTT mediaODvt= optical density of viable tissues incubated in media alone - Evaluation of data The results of the assay was evaluated and compared to negative control. Table: Criteria for in vitro Interpretation: In VitroResults In VivoPredictionMean tissue viability ≤50% Irritant (I), R38Mean tissue viability >50% Non-irritant (NI)- Assay quality controls- Negative Controls (NC)The Dulbecco’s phosphate buffered saline (DPBS) was used as a NC. The assay passed all acceptance criteria if the ODs of the negative control exposed tissues were between ≥0.8 and ≤2.8.  - Positive Controls (PC)5% solution of sodium dodecyl sulfate was used as a PC. The assay is meeting the acceptance criteria if the viability of the PC is ≤20% of the negative control.   - Standard Deviation (SD)The standard deviation (SD) calculated from individual percent tissue viabilities of the test article exposed replicates was ≤18.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 25 mg
- Concentration (if solution): neat

VEHICLE (Not used)
- Amount(s) applied (volume or weight with unit): none
- Concentration (if solution): none- Lot/batch no. (if required): none
- Purity: none

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 30 µL sterile DPBS
- Concentration (if solution): neat

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 30 µL
- Concentration (if solution): 5% solution of sodium dodecyl sulfate

Duration of treatment / exposure:

The exposure times were approximately 1 hour, with ~35 minutes exposure in the incubator and ~25 minutes at room temperature.

Duration of post-treatment incubation (if applicable):

For a total of an approximately 42 hour post-exposure incubation.

Number of replicates:

3 tissues were used for test compound and control.

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

Run 1

Value:

103.9

Vehicle controls validity:

not specified

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: Mean of OD is 2.065; Not irritating

Other effects / acceptance of results:

The MTT data show the assay quality controls were met.

Interpretation of results:

other: Not irritating

Conclusions:

The dermal irritation potential of test article was determined according to the OECD 439 test guideline followed for this study. The Mean % tissue viability compared to negative control (n=3) of the test substance Phenol-formaldehyde resin was determined to be 103.9 %. Thus, substance Phenol-formaldehyde resin was considered to be not irritating to the human skin.

Executive summary:

The dermal irritation potential of test article was determined according to the OECD 439 test guideline for this study. The MatTek EpiDerm™ model was used to assess the potential dermal irritation of the test article by determining the viability of the tissues following exposure to the test article via MTT. Tissues were exposed to the test article and controls for ~one hour, followed by a 42 hour post-exposure recovery period. The viability of each tissue was determined by MTT assay. 

The MTT data show the assay quality controls were met and passed the acceptance of criteria.

The Mean % tissue viability compared to negative control (n=3) of the test substance Phenol-formaldehyde resin (CAS No. 9003 -35 -4) was determined to be 103.9 %.

Hence, under the current experimental test conditions it was concluded that test substance Phenol-formaldehyde resin (CAS No. 9003 -35 -4) was considered to be not irritating to human skin and thus “Not Classified'' as per CLP Regulation.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

secondary literature

Justification for type of information:

Data is from saftey asessment report

Qualifier:

according to guideline

Guideline:

other: as mentioned below

Principles of method if other than guideline:

An eye irritation test was carried out in albino rabbits for test chemical to assess its irritation potential.

GLP compliance:

not specified

Species:

rabbit

Strain:

other: Albino

Details on test animals or tissues and environmental conditions:

No data

Vehicle:

unchanged (no vehicle)

Controls:

yes

Amount / concentration applied:

Amount: 100 mL
concentration applied: 100%

Duration of treatment / exposure:

24 hours

Observation period (in vivo):

7 days

Number of animals or in vitro replicates:

3 (Male/female)

Details on study design:

TEST SITE
- Area of exposure: conjunctival sac of right eye
REMOVAL OF TEST SUBSTANCE
- Washing (if done): The eyes were rinsed with warm isotonic saline solution after 24 hours reading.


-SCORING SYSTEM: The data scored according to the method of Draize et, al.

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

7 d

Score:

10.6

Max. score:

110

Reversibility:

not specified

Remarks on result:

positive indication of irritation

Irritant / corrosive response data:

Slightly irritating effects were observed.

Animal no	Sex	1 hour	24 hour	48 hour	72 hour	120 hour	168 hour
1	Male	8	12	2	0	0	0
2	Female	6	10	0	0	0	0
3	Male	8	10	2	0	0	0
Average		7.3	10.3	1.6	0.0	0.0	0.0

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Conclusions:

The treated animals produced slight ocular reactions with the average score was 10.6 out of maximum score of 110. Hencechemical was considered to be Slightly irritating to the albino rabbits

Executive summary:

An eye irritation test was carried out in 3 albino rabbits for test chemical to assess its irritation potential.

 

100ml undiluted test chemical was installed into theconjunctival sac of right eye of each rabbit.The left eye served as control.The eyes were rinsed with warm isotonic saline solution after 24 hours reading and reactions were scored up to 7 daysaccording to the method of Draize for inflammation.

 

The treated animals produced slight ocular reactions with the average score was 10.6 out of maximum score of 110. Hencechemical was considered to be Slightly irritating to the albino rabbits

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin Irritation

In various studies, the test chemical has been investigated for potential to cause dermal irritation to a greater or lesser extent. The studies are based on in vivo and in vitro experiments in humans, rabbits for the test chemical. The results are summarized as follows:

The dermal irritation potential of test article was determined according to the OECD 439 test guideline for this study. The MatTek EpiDerm™ model was used to assess the potential dermal irritation of the test article by determining the viability of the tissues following exposure to the test article via MTT. Tissues were exposed to the test article and controls for ~one hour, followed by a 42 hour post-exposure recovery period. The viability of each tissue was determined by MTT assay. The MTT data show the assay quality controls were met and passed the acceptance of criteria. The Mean % tissue viability compared to negative control (n=3) of the test substance Phenol-formaldehyde resin (CAS No. 9003 -35 -4) was determined to be 103.9 %. Hence, under the current experimental test conditions it was concluded that test substance Phenol-formaldehyde resin (CAS No. 9003 -35 -4) was considered to be not irritating to human skin and thus “Not Classified'' as per CLP Regulation.

Secondary skin irritation test was conducted in New Zealand Albino rabbits to determine irritating effects of the test chemical. The study was performed according to F.H.S.A Guidelines. 0.5 ml undiluted test chemical was applied to the intact and abraded skin of 6 rabbits for 24 hours. The rabbits were observed for erythema and edema and scored at 4, 24, 48, 72, 168 hours post exposure. The maximum attainable score was 8.

Erythema and edema were observed in all the treated animals at 4 hours which gradually disappeared by 7 days. The mean score after 4 hours was 2.0 and after 7 days was 0.

Since the observed effects and scores were completely reversible by 7 days, the test chemical was considered to be not irritating to skin.

This is supported by results of a Standard patch test conducted on 12 persons worked in factory for test chemical.

They were all exposed to the test chemical on their clothes and on large areas of the skin, particularly the forearms, hands, legs and neck. Eleven had erythema and dryness and some also papules on the forearms, neck, chest and legs. The symptoms decreased during weekends and disappeared spontaneously when they did not work for a week. The dermatitis relapsed within 3 to 4 days at work. Another woman polished the molded, material and her right hand was covered with powder. The skin on the dorsal side and folds between the fingers on the right hand were red and dry.

 

In this patch test, these 12 workers were exposed to 10% the test chemical in petrolatum and were observed for skin reactions.

None of the worker showed positive skin reaction in a standard patch test. Hence, the test chemical was considered to be not irritating in a standard patch test.

These results are supported by a skin irritation test carried out in New Zealand white rabbits for test chemical to assess its irritation potential.

0.5ml of the undiluted test chemical was applied onto the clipped and intact trunk of each rabbit in a one inch by one inch square patch. The patches were held in place with adhesive tape and the trunk of each animal was wrapped with plastic strips to retard evaporation and avoid contamination.

The test sites were exposed for 24 hours and skin reaction were scored up to 7 daysaccording to the method of Draize, Woodart and Calvery.

The treated animals produced slight skin irritation reactions with the average score was 1.3 out of maximum score of 8 after 24 hours of observation. The effects were completely reversible by 7 days with the average score of 0.0.

Based on the scores and observations, the test chemical can be considered to be not irritating to rabbit skin.

The above results are further supported by a study reports a case of accidental exposure to the test chemical over a large area of the skin.

38 year male accidentally spilled a large amount of the test chemical over large cutaneous areas of both his arms and right leg. The spilled chemical was not cleared. 3 days after the event, necrotic lesions appeared over the exposed areas followed by fever, cough, dyspnea and headache. After 4 days of treatment with amoxycillin he was referred to hospital because of continuing deterioration of his condition. Physical examination revealed tachypnea of 28/min, sweating, periorbital and perioral oedema with cyanosis and fever 38.8°C. Blood pressure was 210/120 mmHg, pulse 110/min and regular. Over both lungs diffuse rales and wheezing were noted. A 2/6 systolic murmur was heard over the left sternal border with IV diastolic sound. The skin over the right leg and both arms was oedematous and hard with necrotic and crustate lesions 0.5-2.5 cm in diameter. The rest of the physical examination was normal.  

Subsequently his general and clinical status improved progressively. Tachypnea disappeared and fever subsided on the 5th hospitalization day. Blood pressure returned to normal and skin lesions improved considerably. Chest X-rays returned to normal on the 7th hospitalization day and he was discharged on the 9th day in good general and clinical condition without any treatment.

All symptoms improved progressively and eventually disappeared after 9 days of hospitalization. Hence the test chemical was considered to be not irritating to skin. Also the study suggests that the workers handling the test chemical should be instructed to take appropriate precautions.

In another patch test carried out in 88 workers for test chemical with the primary purpose of conducting a survey of work related skin diseases as a basis for possible preventive measures in a Swedish factory that produces fiber-resin composite.

The investigation was initiated because several workers suffered skin problems on areas of exposed skin when a new technique was introduced at their factory.

Survey of occupational dermatoses, based on a questionnaire, clinical examination, and patch test, was carried out among the workers. The study was approved by the ethical committee at Lund University.

In this patch test, 88 workers were exposed to the test chemical for 2 days and reactions were examined after 1 or 2 additional days, according to International Contact Dermatitis Research Group criteria. A second reading was done 7 days after the initial application of patches.

In six workers, contact allergy to PFR was noted. All six reacted to PFR:2 in the standard series. Three workers reacted to one or more of the different PFRs used in the tests (PFR:X, PFR:Y, and PFR:Z). Occupational dermatitis was diagnosed in nine of 88 (10.2%) investigated persons. In this group, the dermatitis was localized to the lower arms, hands, and face. All nine had allergic contact dermatitis and were not diagnosed with irritant contact dermatitis. Non-work related dermatoses were diagnosed in 18 (20.5%) study subjects. Examples of non-work-related skin diagnoses were acne vulgaris, atopic eczema, psoriasis, and seborrhoic eczema.

Because of these skin problems, the plant abandoned the new technique and went back to the traditional application, and by doing that, most workers got well. It was observed that the phenol -formaldehyde resin used contained fairly high concentrations of sensitizing monomers and dimmers.

On the basis of all the observed effects it was considered that the test chemical was irritating to the human skin.

Even though results of an in vivo experimental data claim that the test chemical caused irritation to human skin. But the results of the remaining studies including in vitro studies give strong evidence that the test chemical may indeed be not irritating to skin.

Hence, the test chemical can be considered to be not irritating to skin. Comparing the above annotations with the criteria of CLP regulation, the test chemical was classified under the category “Not Classified”

Eye Irritation

In various studies, the test chemical has been investigated for potential to cause ocular irritation to a greater or lesser extent. The studies are based on in vivo experiments in rabbits for the test chemical. The results are summarized as follows:

An eye irritation test was carried out in 3 albino rabbits for test chemical to assess its irritation potential.

100ml undiluted test chemical was installed into the conjunctival sac of right eye of each rabbit. The left eye served as control. The eyes were rinsed with warm isotonic saline solution after 24 hours reading and reactions were scored up to 7 days according to the method of Draize for inflammation.

The treated animals produced slight ocular reactions with the average score was 10.6 out of maximum score of 110. Hence, chemical was considered to be Slightly irritating to the albino rabbits.

This is supported by the results of an Acute eye irritation test conducted for test chemical in 6 New Zealand Albino rabbits to determine the its adverse ocular effects.

0.1ml undiluted test chemical was placed into the eye of each rabbit for 24 hours and reactions were scored for 7 days.

Areas of barely perceptible to slight corneal cloudiness, iris showed little or no reaction to light, sever erythema very slight to slight edema, copious discharge containing whitish exudates was observed. The average mean eye irritation score of the test chemical was 51.4 (maximum score of 110).

Since the chemical was able to elicit ocular lesions, the test chemical was considered to be moderately irritating to the eyes of New Zealand White rabbits.

The above results are further supported by an ocular irritation test was conducted for test chemical in New Zealand Albino rabbits to assess its irritation potential.

About 100mg of the test chemical (free phenol: 26.6%) was installed into conjunctival sac of one of the eye of each rabbit and other eye served as control. The animals were observed for at least 7 and in some cases up to 21 days, or until all signs of irritant reactions had resolved. Ocular lesions were examined at1, 24, 48 and 72 hours and 7 days after installation using the numerical system developed by Draize.

Severe irritation and tissue damage with extensive corneal opacity was observed. Animals were killed to avoid unnecessary suffering. Also the chemical was observed to be corrosive to the eyes of each rabbit. Hence the test chemical was considered to be severely irritating to the eyes of three female New Zealand White rabbits.

The results from the available studies indicate a very strong possibility that the test chemical can be indeed irritating to rabbit eyes. Hence, it can be classified under the category “Category 2” as per CLP regulation.

Justification for classification or non-classification

Available results for the test chemical indicate a strong possibility that the test chemical lacks the potential to cause irritation to skin. Hence, the test chemical can be considered to be not irritating to skin and classified under the category “Not Classified” as per CLP regulation.

Available results for the test chemical indicate a strong possibility that the test chemical has the potential to cause irritation to eyes. Hence, the test chemical can be considered to be irritating to eyes and classified under the category “Category 2” as per CLP regulation.