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Administrative data

Description of key information

Oral toxicity: LD50 of 674 mg/kg (based on Malachite Green; salt form not specified)

Dermal toxicity: LD0 of 2000 mg/kg b.w (based on Malachite Green Oxalate)

Inhalatory exposure is unlikely

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
Endpoint:
acute toxicity: oral
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Well described, no GLP and no international guideline followed. Read across from a similar substance which has the same main component and with a different counter ion that doesn't influence the characteristics related to the specific end-point
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
not specified
GLP compliance:
no
Test type:
acute toxic class method
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Winkelmann, Borchen
- Age at study initiation: 98 days old
- Number of animals: 9 males, 14 females
- Weight at study initiation: Males 165 -184 g and females 160-182 g
- Diet: pellet Alrtomin R 1324 ad libitum
- Water: Tap water ad libitum
- Other: all animals were marked into skin by picric acid

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 1.5 °C
- Humidity (%): 60 ± 5%
- Photoperiod: 12 H radk/light cycle, artificial light

Route of administration:
oral: gavage
Vehicle:
not specified
Details on oral exposure:
VEHICLE: oral, gavage
Doses:
0.5, 0.8, 1.3, 1.6, 2.0 ml/kg/bw
No. of animals per sex per dose:
5 animal x each sex x dose
Control animals:
yes
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: twice a day
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,
Statistics:
The calculation of the LD-50 for the confidence interval p <0.05 was conducted in accordance with to Rosier et al.J. Tox. Environ. Health 3, 1977, 797th
Sex:
male/female
Dose descriptor:
LD50
Effect level:
ca. 1.2 mL/kg bw
Based on:
test mat.
95% CL:
> 1.1 - < 1.4
Remarks on result:
other: Estimated at 100 %: LD50 = ca 674 mg/Kg bw
Mortality:
All animals are dead by a dose of 5 ml/kg bw. The deaths were within the first 6 days after the application registered.
Clinical signs:
Dose levels between 0.8 to 2.0 ml/kg: in all the animals worsening health and bristling.
At doses ≥ 1.3 ml/kg:: manifest signs of relaxation and abdominal numbness.
At doses ≥ 0.8 ml / kg: growth retardation.
The symptoms occurred about 1 to 2 hours after application and kept in individual animals until the end of the experiment.

Results are expressed as ml/kg; correcting the value following the sample density indicated on the original report (1.123 g/mL) result is LD50 = 674 mg/kg bw

Group Dose [ml/kg bw] Result  Symptoms duration  Dead time Mortality
Males and females
1 0.5 0/0/10 -- -- 0
2 0.8 2/10/10 24h - 14d 3d - 6d 20
3 1.3 5/10/10 2h - 7d 4h - 5d 50
4 1.6 7/10/10 1h - 14d 2h - 3d  70
5 2.0  10/10/10 1h - 2d 2h - 3d 100
Interpretation of results:
other: CLP: Acute tox 4
Remarks:
Criteria used for interpretation of results: EU
Conclusions:
Estimated at 100 %: LD50 = ca 674 mg/Kg bw
Executive summary:

Acute oral toxicity test with MS Astrazon Gruen liquid 50% by administration to gavage on male and female wistar rats.

Estimated at 100 %: LD50 = ca 674 mg/Kg bw

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
674 mg/kg bw

Acute toxicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
no study available

Acute toxicity: via dermal route

Link to relevant study records
Reference
Endpoint:
acute toxicity: dermal
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Not GLP study, aged study Read across from a similar substance which has the same main component and with a different counter ion that doesn't influence the characteristics related to the specific end-point
Reason / purpose:
reference to same study
Principles of method if other than guideline:
For skin application a site of 35 cm^2 was clipped 3 h before administration and the site covered by four-ply gauze and an elastic bandage.
GLP compliance:
no
Test type:
standard acute method
Limit test:
yes
Species:
other: rat and guinea pig
Strain:
other: Wistar and Ssc
Sex:
not specified
Details on test animals and environmental conditions:
TEST ANIMALS
RATS
- Strain: Wistar rats, Mol: WIST (SPF)
- Weight at study initiation: 170-250 g
- Source: from Molllegard Breeding Centre (L1. Skensved, Denmark)

GUINEA PIG
- Strain: Ssc, AL
- Source: Statens Serumininstitut, (Copenhagen, Denmark)
- Weight at study initiation: 300 g

All were given food and water freely except 16 h before gavage.
For rats
- Temperature: kept at 22 ± 1°C
- Humidity: 60 ± 10%
- Light: from 21.00 to 09.00 with air changes eight times/h.
Type of coverage:
occlusive
Vehicle:
water
Details on dermal exposure:
TEST MATERIAL
- Amount(s) applied: Aqueous solution
- Concentration: MGH at 20%
Duration of exposure:
14 days
No. of animals per sex per dose:
Groups of five animals of either sex
Control animals:
yes, concurrent no treatment
Details on study design:
For skin application a site of 35 cm^2 was clipped 3 h before administration and the site covered by four-ply gauze and an elastic bandage (Acrylastic, Beiersdorf AG, FRG)

Observation period: 1, 2, 3, and 5 h after dosing and each day for 14 days
Sex:
male/female
Dose descriptor:
LD0
Effect level:
ca. 2 000 mg/kg bw
Based on:
test mat.

No signs of systemic toxicity were observed after occlusive dermal application of 2000 mg/kg. As a 20% suspension of Malachite Green did not produce visible erythema or oedema on either rats or guinea pigs; authors hadn't deemed it meaningful doing a rabbit skin irritation study. Similarly, within the conditions of the guinea pig maximisation test no effects could be seen on controls or treated animals.

Interpretation of results:
other: CLP: not classified
Remarks:
Criteria used for interpretation of results: EU
Conclusions:
Acute Dermal Toxicity: no signs of systemic toxicity were observed after occlusive dermal application of 2000 mg/kg
Executive summary:

Rats ang guinea pig were tested for dermal acute toxicity. For skin application a site of 35 cm^2 was clipped 3 h before administration and the site covered by four-ply gauze and an elastic bandage.

No signs of systemic toxicity were observed after occlusive dermal application of 2000 mg/kg. As a 20% suspension of Malachite Green did not produce visible erythema or oedema on either rats or guinea pigs; authors hadn't deemed it meaningful doing a rabbit skin irritation study. Similarly, within the conditions of the guinea pig maximisation test no effects could be seen on controls or treated animals.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating dose
Value:
2 000 mg/kg bw

Additional information

All studies assess acute oral toxicity only with Malachite Green Oxalate.

LD50 of 674 mg/kg has been chosen as the most reliable data because the study was complete and good conducted, with all tables and graphics reported (Dystar, 1983).

Results of Dystar study is coherent with results reported in other studies.

The outcomes from Clemmensen et al (1984) report were previously accepted to classify Malachite Green (MG) for oral acute toxicity; this classification is today not confirmed by Dystar studies. Clemmensen study does not report any details and information on test conditions.

In the study of Clemmensen et al, (1984) it is reported that no systemic effects were seen after dermal application of 2000 mg/kg b.w.

Inhalator exposure is unlikely; nevertheless in a study on teratogenical effects on rabbits (Meyer and Jorgenson, 1983) the authors reported that if MG during gavage was coughed up and aspirate into the lungs, those animals died within minutes of acute pulmonary toxicity.

Justification for selection of acute toxicity – oral endpoint

Most conservative and reliable data.

Justification for classification or non-classification

The results form reliable study do not confirm the outcomes of Clemmensen, that were previously accepted to classify Malachite Green Acetate for oral acute toxicity.

Nevertheless, for the time being the harmonised classification will need to be implemented, therefore

According to CLP regulation (EC1272/2008) Malachite Green Acetate is classified as Acute toxicity 4, H302