Registration Dossier

Administrative data

Description of key information

Skin Irritation:

Basic Violet 3 failed to produce any dermal reactions in the rabbits.

Hence, Basic Violet 3 was considered to be not irritating to rabbit skin.

Eye Irritation:

Basic Violet 3 produced severe and persistent blepharitis with hyperaemia, oedema and necrosis of the conjunctivae and nictitating membrane. At 3 weeks there was gross opacification, deformity and vascularization of the cornea.

Since the effects were irreversible even after 21 days, Basic Violet 3 can be considered highly irritating to rabbit eyes.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Justification for type of information:
data is from experimental reports following standard procedures
Qualifier:
according to
Guideline:
other: as mentioned below
Principles of method if other than guideline:
To assess the dermal irritation potential of Basic Violet 3 in rabbits
GLP compliance:
not specified
Specific details on test material used for the study:
- Name of test material (as cited in study report): Crystal violet
- IUPAC name: N-(4-{bis[4-(dimethylamino)phenyl]methylene}cyclohexa-2,5-dien-1-ylidene)-N-methylmethanaminium chloride
- Molecular formula: C25H30N3.Cl
Molecular weight: 407.986 g/mol
- Smiles notation: C(\c1ccc(N(C)C)cc1)(c1ccc(N(C)C)cc1)=C1\C=C\C(=[N+](/C)C)C=C1.[ClH-]
- InChl:1S/C25H30N3.ClH/c1-26(2)22-13-7-19(8-14-22)25(20-9-15-23(16-10-20)27(3)4)21-11-17-24(18-12-21)28(5)6;/h7-18H,1-6H3;1H/q+1;/p-1
- Substance type: Organic
- Physical state: Solid
Species:
rabbit
Strain:
other: albino rabbits
Details on test animals and environmental conditions:
sex: male
Type of coverage:
semiocclusive
Preparation of test site:
clipped
Vehicle:
unchanged (no vehicle)
Controls:
not specified
Amount / concentration applied:
no data available
Duration of treatment / exposure:
24 hours
Observation period:
observations were made just after patch removal and also at 48 hours
Number of animals:
6 male albino rabbits
Details on study design:
TEST SITE
- Area of exposure: trunk and lateral areas
- % coverage: one inch sqaures of test material were applied to intact skin under 1inch*1inch gauze squares (double thickness)
- Type of wrap if used: rubber sheeting was loosely wrapped around the trunk and secured with an adhesive tape

REMOVAL OF TEST SUBSTANCE
- Washing (if done): no


OBSERVATION TIME POINTS
(indicate if minutes, hours or days) : observations were made just after patch removal and also at 48 hours

SCORING SYSTEM:
- Method of calculation: the observations were graded according to the system of the regulations of the Federal Hazardous Substances Act.
Other effects / acceptance of results:
no data available
Irritation parameter:
overall irritation score
Basis:
mean
Time point:
48 h
Reversibility:
not specified
Remarks on result:
no indication of irritation
Irritant / corrosive response data:
the test material failed to produce any dermal reactions in the rabbits
Interpretation of results:
other: not irritating
Conclusions:
Basic Violet 3 failed to produce any dermal reactions in the rabbits.
Hence, Basic Violet 3 was considered to be not irritating to rabbit skin.
Executive summary:

The dermal irritation potential of Basic Violet 3 was assessed in rabbits. 6 male albino rabbits were clipped free of hair on the trunk and lateral areas and placed in a FDA- type stocks. One inch sqaures of test material were applied to intact skin under 1inch*1inch gauze squares (double thickness). Rubber sheeting was loosely wrapped around the trunk and secured with an adhesive tape. After 24 hours, the rabbits were removed from the stocks and reactions were graded. Observations were also made at 48 hours. The observations were graded according to the system of the regulations of the Federal Hazardous Substances Act.

Basic Violet 3 failed to produce any dermal reactions in the rabbits.

Hence, Basic Violet 3 was considered to be not irritating to rabbit skin.

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:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
data is from peer reviewed journals
Qualifier:
according to
Guideline:
other: Standard Eye irritation test
Principles of method if other than guideline:
The effects of Basic Violet 3 on the rabbit eye were investigated using a standard rabbit eye irritation test
GLP compliance:
not specified
Specific details on test material used for the study:
- Name of test material (as cited in study report): Basic Violet 3
- IUPAC name: N-(4-{bis[4-(dimethylamino)phenyl]methylene}cyclohexa-2,5-dien-1-ylidene)-N-methylmethanaminium chloride
- Molecular formula : C25H30N3.Cl
Molecular weight : 407.986 g/mol
- Smiles notation: C(\c1ccc(N(C)C)cc1)(c1ccc(N(C)C)cc1)=C1\C=C\C(=[N+](/C)C)C=C1.[ClH-]
- InChl:1S/C25H30N3.ClH/c1-26(2)22-13-7-19(8-14-22)25(20-9-15-23(16-10-20)27(3)4)21-11-17-24(18-12-21)28(5)6;/h7-18H,1-6H3;1H/q+1;/p-1
- Substance type: Organic
- Physical state: Solid
Species:
rabbit
Strain:
not specified
Details on test animals or tissues and environmental conditions:
no data available
Vehicle:
water
Controls:
not specified
Amount / concentration applied:
20mg/ml
Duration of treatment / exposure:
single exposure
Observation period (in vivo):
till 21 days
Duration of post- treatment incubation (in vitro):
no data available
Number of animals or in vitro replicates:
no data available
Details on study design:
no data available
Other effects / acceptance of results:
no data available
Irritation parameter:
overall irritation score
Basis:
mean
Time point:
21 d
Reversibility:
not fully reversible within: 21 days
Remarks on result:
probability of severe irritation
Irritant / corrosive response data:
It produced severe and persistent blepharitis with hyperaemia, oedema and necrosis of the conjunctivae and nictitating membrane.Mild keratitis was apparent within 24 h, and over the ensuing 21 days became progressively more severe. At 3 weeks there was gross opacification, deformity and vascularization of the cornea. Keratitis often obscured a severe iritis.
Interpretation of results:
Category 1 (irreversible effects on the eye) based on GHS criteria
Conclusions:
Basic Violet 3 produced severe and persistent blepharitis with hyperaemia, oedema and necrosis of the conjunctivae and nictitating membrane. At 3 weeks there was gross opacification, deformity and vascularization of the cornea.
Since the effects were irreversible even after 21 days, Basic Violet 3 can be considered highly irritating to rabbit eyes.
Executive summary:

The effects of Basic Violet 3 on the rabbit eye were investigated using a standard rabbit eye irritation test. 20 mg/ml of Basic Violet 3 in water was instilled into the eyes of rabbits and observed till 21 days. Basic Violet 3 produced severe and persistent blepharitis with hyperaemia, oedema and necrosis of the conjunctivae and nictitating membrane. Mild keratitis was apparent within 24 h, and over the ensuing 21 days became progressively more severe. At 3 weeks there was gross opacification, deformity and vascularization of the cornea. Keratitis often obscured a severe iritis.

Since the effects were irreversible even after 21 days, Basic Violet 3 can be considered highly irritating to rabbit eyes.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irreversible damage)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin Irritation:

Various studies for the target substance were summarized as follows:

The dermal irritation potential of Basic Violet 3 was assessed (Haskell Laboratory for Toxicology and Industrial medicine, Sustainability Support Services (Europe) AB has letter of access, 1974) in rabbits. 6 male albino rabbits were clipped free of hair on the trunk and lateral areas and placed in a FDA- type stocks. One inch squares of test material were applied to intact skin under 1inch*1inch gauze squares (double thickness). Rubber sheeting was loosely wrapped around the trunk and secured with an adhesive tape. After 24 hours, the rabbits were removed from the stocks and reactions were graded. Observations were also made at 48 hours. The observations were graded according to the system of the regulations of the Federal Hazardous Substances Act.

Basic Violet 3 failed to produce any dermal reactions in the rabbits. Hence, Basic Violet 3 was considered to be not irritating to rabbit skin.

This result is supported by the experimental study summarized in Scientific Committee on Cosmetology (seventh series), 1988; for Basic Violet 3. 500mg of undiluted Basic Violet 3 was applied to rabbit skin and later observed for skin reactions (duration not specified).

Since, Basic Violet 3 failed to induce any skin reaction, it was considered to be not irritating to the rabbit’s skin.

These results are further supported by the experimental studies performed by Håkan Mobacken(The Department of Dermatology, University of Göteborg, Göteborg, Sweden, 1974) to assess the dermal irritation potential of Basic Violet 3 in guinea pigs and humans.

5 guinea pigs were stripped on the flank and immediately painted with 2% gentian violet (crystal violet). Skin biopsies were taken after 3 days in guinea pigs. The dye was applied to the intact skin and observed for signs of irritation. All guinea pigs developed necrotic reactions after 2 days of stripping, but only in the treated sites. No reaction was observed when the dye was applied to intact skin. These observations indicate that Crystal violet under certain conditions (stripping) can cause severe dermal irritation to guinea pig skin.

Similar observations were reported in humans. 2 human volunteers were stripped on the flexor surface of the forearm and immediately painted with 1% gentian violet (crystal violet) in water once a day for 2 days on half of the stripped area. The other half served as control. Skin biopsies were taken from the stripped areas after 1 and 5 days in man. The dye was applied to the intact skin and observed for signs of irritation. In man ulcerations were seen after 5 days in areas painted with crystal violet. No reaction was observed when the dye was applied to intact skin. These observations indicate that Crystal violet under certain conditions (stripping) can cause severe dermal irritation to human skin.

Hence, it can be considered that Crystal Violet is not irritating to intact skin but causes necrosis when applied to stripped or wounded skin.

Based on the available data for the target,Basic Violet 3 can be considered to be not irritating to skin.Comparing the above annotations with the criteria of CLP regulation, Basic Violet 3 can be classified under the category “Not Classified”

 

Eye Irritation:

Various studies for the target substance were summarized as follows:

The effects of Basic Violet 3 on the rabbit eye were investigated by Ballantyne B et.al (British Journal of Pharmacology. 1973;49(1):181P-182P) using a standard rabbit eye irritation test. 20 mg/ml of Basic Violet 3 in water was instilled into the eyes of rabbits and observed till 21 days. Basic Violet 3 produced severe and persistent blepharitis with hyperaemia, oedema and necrosis of the conjunctivae and nictitating membrane. Mild keratitis was apparent within 24 h, and over the ensuing 21 days became progressively more severe. At 3 weeks there was gross opacification, deformity and vascularization of the cornea. Keratitis often obscured a severe iritis.

Since the effects were irreversible even after 21 days, Basic Violet 3 can be considered highly irritating to rabbit eyes.

The eye irritation potential of Basic Violet 3 was assessed (Haskell Laboratory for Toxicology and Industrial medicine, Sustainability Support Services (Europe) AB has letter of access, 1972) in rabbits. 10 mg of undiluted powder [H7369, H7371, H7372] was instilled in the right conjunctival sac of 2 albino rabbits for each powder. Basic violet 3 was also tested by instilling 0.1ml as 10% aqueous solution[H7369,7371,7372] and  0.1ml as undiluted product [H7370] in the right conjunctival sac of 2 additional albino rabbits/ sample. After 20 seconds of contact, one treated eye of each pair was washed with tap water for one minute. The other exposed eye was not washed. The eyes were observed at 1,4 hours and at 1,2,3,4,7 and 14 days for signs of irritation. Observations of the iris, cornea and conjunctivae were made with a hand-slit lamp at 1,4 hours and at 1,2,3,4,7 and 14 days  if necessary. A biomicroscope was used at examination after the day of treatment. 5% aqueous fluorescein stain was also used at examinations after day of treatment. Since the undiluted Basic Violet 3 caused severe irritation which did not subside by 14 days, Basic Violet 3 can be considered to be severely irritating to rabbit eyes.

Even though Basic Violet 3[H 7369, H7371, H7372] produced mild to transient irritation which was reversible in 3 days when instilled as a solid powder and as 10% aqueous solution, the undiluted Basic Violet 3[H7370] caused severe irritation which did not subside by 14 days. Hence, Basic Violet 3 can be considered to be severely irritating to rabbit eyes.

The above results are also supported by the experimental study performed by SP Dhir et.al(Indian J Ophthalmol, 1982;30:21-22) to assess the irritation potential of Basic Violet 3 in rabbits.

1% gentian violet[Basic Violet 3]was instilled in conjunctival sac of 2 rabbits thrice in a day. The histopathological appearance of normal conjunctiva obtained from control rabbit was compared with the treated eyes. Both the rabbits developed conjunctival congestion and discharge next day. After 3 days of instillation there was some necrosis of the conjunctiva. Conjunctival biopsies obtained from affected rabbits showed variable thinning of epithelial lining with total loss of goblet cells and subepithelial capillary congestion with neutrophilic infiltration.In experimental studies on rabbits, gentian violet [Basic Violet 3] caused gross destruction of goblet cells and thinning of the conjunctival epithelium. Hence, Basic Violet 3 can be considered to be a severe eye irritant.

SP Dhir et.al(Indian J Ophthalmol, 1982;30:21-22) reported an case of accidental exposure to Gentian Violet[Basic Violet 3] to human eyes and its effects. A 60- year-old male had 1% Gentian Violet accidentally instilled into both his eyes by a medical practitioner. The patient complained of irritation, pain, and diminution of vision. On examination, his visual acuity was reduced to counting fingers ½ meter OD (right eye) and 6/36 OS (left eye). Moderate lid edema and blepharospasm were present. The conjunctivae were congested and chemosed. Corneas were hazy and edematous in both eyes. Biomicroscopic examination revealed punctered epithelial lesions scattered all over the cornea. At the end of 5 weeks, the visual acuity had improved to counting fingers at 1 meter OD and 6/9 OS. The marginal tear strip was absent. Tear breakup time was 5 seconds in both eyes. After 4 months, the tear secretion had not shown any improvement. Basic Violet 3 led to the development of keratoconjunctivitis sicca which did not show any improvement during the follow up period. Since the effects were irreversible even after 4 months, Basic Violet 3 can be considered highly irritating to eyes.

These studies are further supported by the experimental study summarized in Scientific Committee on Cosmetology (seventh series), 1988; for Basic Violet 3. 0.25% suspension in saline and higher concentrations were instilled into the rabbit eyes and observed for effects (number of animals and duration not specified). Corneal turbidity and swelling of the iris and conjunctivae were observed. When exposed to higher concentrations, necrosis of the iris and severe swelling of the conjunctivae was observed.

Based on these observations, Basic Violet 3 can be considered to be highly irritating to rabbit eyes.

Since majority of the studies align towards the possibility of Basic Violet 3 being highly irritating to eyes causing irreversible damage, though some results suggest that Basic Violet 3 may not cause severe irritation to eyes. Hence, by applying the weight of evidence approach,Basic Violet 3 can be considered to be highly irritating to eyes causing irreversible damage.Comparing the above annotations with the criteria of CLP regulation, Basic Violet 3 can be classified under the category “Category 1”.

Justification for classification or non-classification

Available information for Basic Violet 3 indicates that it is not likely to cause any irritation to skin, but is highly irritating to eyes causing irreversible damage.

Hence, Basic Violet 3 can be classified under the category “Not Classified” for skin irritation and “Category 1” for eye irritation as per CLP regulation.