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Acute Toxicity: inhalation

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Administrative data

Endpoint:
acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1989
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
REPORTING FORMAT FOR THE ANALOGUE APPROACH

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
Target and source differ only in the salt form

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
Source: Mixture of Basic Violet 16 Oxalate / EC 281-588-8 / CAS 83969-11-3 and Basic Violet 16 Sulfate / EC 281-474-8 / CAS 83950-22-5
Target: Basic Violet 16 Dihydrogenphosphate / EC 278-248-6 / CAS 75535-16-9

3. ANALOGUE APPROACH JUSTIFICATION
see attached justification document in section 13
Cross-reference
Reason / purpose:
read-across source
Reference
Endpoint:
acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1989
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Qualifier:
according to
Guideline:
EU Method B.2 (Acute Toxicity (Inhalation))
GLP compliance:
yes
Test type:
standard acute method
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Winkelmann, Borchen, Germany
- Age at study initiation: 2 to 3 months
- Weight at study initiation: 160 g to 200 g
- Fasting period before study: -
- Housing: groups of 5
- Bedding: wood glanulate type S 8/15 Ssniff, Germany
- Diet (ad libitum): ltrominc)1324 - Haltungsdiat fur Ratten und Mause, Altromin GmbH, Lage, Germany
- Water (ad libitum): tap
- Acclimation period: at least 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24
- Humidity (%): ca. 50
- Air changes (per hr): ca. 10-fold
- Photoperiod (hrs dark / hrs light): 12/12


IN-LIFE DATES: From: 8. Sep to 28. Oct 1988
Route of administration:
inhalation: dust
Type of inhalation exposure:
nose/head only
Vehicle:
air
Mass median aerodynamic diameter (MMAD):
4 µm
Geometric standard deviation (GSD):
1.94
Remark on MMAD/GSD:
8.0 mg/m³ air: MMAD: 3.98 µm; GSD: 1.94
53 mg/m³ air: MMAD: 3.04 µm; GSD: 2.21
240 mg/m³ air: MMAD: 5.58 µm; GSD: 1.54
2386 mg/m³ air: MMAD: 32.1 µm; GSD: 2.11
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: exposure tubes were adapted to the growth of the rats (Fa Rhema Labortechnik, Hofheim, Germany) - Head/Nose exposition
- Exposure chamber volume: ca. 20 liter
- Source and rate of air: 2 parallel connected Boge compressors (SB 270/15/350D) - ca. 800 to 1000 kPa air pressure; 84-times air change rate/h, monitored by a calibrated Rotameter
- Method of conditioning air: conditioned by a downstream "VIA-Drucklufttrockner" (Typ A 110)
- System of generating particulates/aerosols: High concentrations: "Bayer-Staubgeneratoren"; low concentrations: "Wright-Dust-Feeder"; nebulized in cylindric inhalation chamber under dynamic conditions
- Method of particle size determination: gravimetric with Andersen cascade impactor
- Treatment of exhaust air: 70% of the fed air is exhaust air, purified by a cotton filter
- Temperature & humidity in air chamber (Leybold-Heraeus Meßsystem):
- without rats
- temperature: 24°C
- rel. humidity: 14%
- with rats
- temperature: 25°C
- rel. humidity: 34%
- during recent study
- temperature: 20 - 24°C
- rel. humidity: < 10%
- Pressure in air chamber:
- max. dispersion pressure: ca. 200 kPa
- supply air: ca. 28 L/min
- exhaust air: ca. 20 L/min


TEST ATMOSPHERE
- Brief description of analytical method used: filter analysis, gravimetrical determination
- Samples taken from breathing zone: yes

VEHICLE
- Composition of vehicle (if applicable): no vehicle other than air used
- Concentration of test material in vehicle (if applicable): 8, 53, 240, 2386 mg/m³ air


TEST ATMOSPHERE (if not tabulated)
- MMAD (Mass median aerodynamic diameter) / GSD (Geometric st. dev.):
8.0 mg/m³ air: MMAD: 3.98 µm; GSD: 1.94
53 mg/m³ air: MMAD: 3.04 µm; GSD: 2.21
240 mg/m³ air: MMAD: 5.58 µm; GSD: 1.54
2386 mg/m³ air: MMAD: 32.1 µm; GSD: 2.11
Analytical verification of test atmosphere concentrations:
yes
Duration of exposure:
4 h
Concentrations:
target concentration: 8.0 mg/m³ air (all concentrations are given in mg dye/m³ air)
computed concentration: 13.1 mg/m³ air
MMAD: 3.98 µm; GSD: 1.94; NMAD: 1.06 µm; SMAD: 2.56 µm
respirability (% <= 5 µm): mass related: 64% (measured)
number related: 99% (extrapolated)

target concentration: 53 mg/m³ air
computed concentration: 47.5 mg/m³ air
MMAD: 3.04 µm; GSD: 2.21; NMAD: 0.458 µm; SMAD: 1.62 µm
respirability (% <= 5 µm): mass related: 74% (measured)
number related: 100% (extrapolated)

target concentration: 240 mg/m³ air
computed concentration: 209.2 mg/m³ air
MMAD: 5.58 µm; GSD: 1.54; NMAD: 3.15 µm; SMAD: 4.61 µm
respirability (% <= 5 µm): mass related: 41% (measured)
number related: 86% (extrapolated)

target concentration: 2386 mg/m³ air
computed concentration: 6240.6 mg/m³ air
MMAD: 32.1 µm; GSD: 2.11; NMAD: 5.97 µm; SMAD: 18.3 µm
respirability (% <= 5 µm): mass related: 1% (measured)
number related: 41% (extrapolated)
No. of animals per sex per dose:
5
10 controls
Control animals:
yes
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
- clinical signs: Day 1: multiple times
Days 2 to 14: twice daily
- body weight: pre-treatment, Day 3, Day 7, weekly thereafter
- Necropsy of survivors performed: yes
- other: Irwin screen, rectal temperature
Statistics:
means , standard deviations, statistical significance (p<=0.05 and 0.01)

Necropsy: Respiratory tract findings: RxC Chi-Square test, pairwise Fisher's test
Body weight: means , standard deviations; analysis of variances; Box-test for homogeneity; Games and Howell modification of Tukey-Kramer tesst
Particle analysis: MMAD, GSD (probit-transformed mass-related cumulative frequency distribution, logarithmized effective cut-off diameter
Concentration analytics: means, standard deviations
LC50: Maximum-Likelihood method
Key result
Sex:
male/female
Dose descriptor:
LC50
Effect level:
53 mg/L air (analytical)
Based on:
act. ingr.
Exp. duration:
4 h
Sex:
male/female
Dose descriptor:
LC50
Effect level:
0.053 mg/L air (analytical)
Based on:
act. ingr.
Exp. duration:
4 h
Mortality:
Conc. [mg/m³] deaths (M/F)
8 0/0
53 2/4
240 5/5
2386 5/5
Clinical signs:
dose dependent clinical signs
all dose-groups: hypoactivity, piloerection, unkempt appearance, labored respiration, bradypnoea

in addition at
53 mg/m³: agonal tonic convultions - males
240 mg/m³: agonal ataxia - all
agonal: apnoea, prone position, coma - males
2386 mg/m³: all animals died during eyposure
Body weight:
transient decrease in surviving animals
Gross pathology:
intermittent deaths: hepatization of lung; lung edema, hydrothorax; pale liver with accentuated lobular pattern; pale mottled kidneys with reddened pelvic area; dark spleen; reddened glandular stomach with ulcerous foci; small intestines reddened, reddish content; gastro-intestinal tract with yellow mucoid content; test-item related discolouration of inner organs.

terminal sacrifices: no effects
Interpretation of results:
Category 2 based on GHS criteria
Conclusions:
Basic Violet 16 showed inhalation toxicity in the rat. This effect was mainly triggered by strong irritatng effects to the lung. There were signs of a concentration-dependent severe irritation of the respiratory tract as well as unspecific clinical signs and a transient decrease in body weight in all surviving animals.
The approximative LC50 for rats is 53 mg/m³ air

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1989
Report Date:
1989

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Qualifier:
according to
Guideline:
EU Method B.2 (Acute Toxicity (Inhalation))
GLP compliance:
yes
Test type:
standard acute method
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Winkelmann, Borchen, Germany
- Age at study initiation: 2 to 3 months
- Weight at study initiation: 160 g to 200 g
- Fasting period before study: -
- Housing: groups of 5
- Bedding: wood glanulate type S 8/15 Ssniff, Germany
- Diet (ad libitum): ltrominc)1324 - Haltungsdiat fur Ratten und Mause, Altromin GmbH, Lage, Germany
- Water (ad libitum): tap
- Acclimation period: at least 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24
- Humidity (%): ca. 50
- Air changes (per hr): ca. 10-fold
- Photoperiod (hrs dark / hrs light): 12/12


IN-LIFE DATES: From: 8. Sep to 28. Oct 1988

Administration / exposure

Route of administration:
inhalation: dust
Type of inhalation exposure:
nose/head only
Vehicle:
air
Mass median aerodynamic diameter (MMAD):
4 µm
Geometric standard deviation (GSD):
1.94
Remark on MMAD/GSD:
8.0 mg/m³ air: MMAD: 3.98 µm; GSD: 1.94
53 mg/m³ air: MMAD: 3.04 µm; GSD: 2.21
240 mg/m³ air: MMAD: 5.58 µm; GSD: 1.54
2386 mg/m³ air: MMAD: 32.1 µm; GSD: 2.11
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: exposure tubes were adapted to the growth of the rats (Fa Rhema Labortechnik, Hofheim, Germany) - Head/Nose exposition
- Exposure chamber volume: ca. 20 liter
- Source and rate of air: 2 parallel connected Boge compressors (SB 270/15/350D) - ca. 800 to 1000 kPa air pressure; 84-times air change rate/h, monitored by a calibrated Rotameter
- Method of conditioning air: conditioned by a downstream "VIA-Drucklufttrockner" (Typ A 110)
- System of generating particulates/aerosols: High concentrations: "Bayer-Staubgeneratoren"; low concentrations: "Wright-Dust-Feeder"; nebulized in cylindric inhalation chamber under dynamic conditions
- Method of particle size determination: gravimetric with Andersen cascade impactor
- Treatment of exhaust air: 70% of the fed air is exhaust air, purified by a cotton filter
- Temperature & humidity in air chamber (Leybold-Heraeus Meßsystem):
- without rats
- temperature: 24°C
- rel. humidity: 14%
- with rats
- temperature: 25°C
- rel. humidity: 34%
- during recent study
- temperature: 20 - 24°C
- rel. humidity: < 10%
- Pressure in air chamber:
- max. dispersion pressure: ca. 200 kPa
- supply air: ca. 28 L/min
- exhaust air: ca. 20 L/min


TEST ATMOSPHERE
- Brief description of analytical method used: filter analysis, gravimetrical determination
- Samples taken from breathing zone: yes

VEHICLE
- Composition of vehicle (if applicable): no vehicle other than air used
- Concentration of test material in vehicle (if applicable): 8, 53, 240, 2386 mg/m³ air


TEST ATMOSPHERE (if not tabulated)
- MMAD (Mass median aerodynamic diameter) / GSD (Geometric st. dev.):
8.0 mg/m³ air: MMAD: 3.98 µm; GSD: 1.94
53 mg/m³ air: MMAD: 3.04 µm; GSD: 2.21
240 mg/m³ air: MMAD: 5.58 µm; GSD: 1.54
2386 mg/m³ air: MMAD: 32.1 µm; GSD: 2.11
Analytical verification of test atmosphere concentrations:
yes
Duration of exposure:
4 h
Concentrations:
target concentration: 8.0 mg/m³ air (all concentrations are given in mg dye/m³ air)
computed concentration: 13.1 mg/m³ air
MMAD: 3.98 µm; GSD: 1.94; NMAD: 1.06 µm; SMAD: 2.56 µm
respirability (% <= 5 µm): mass related: 64% (measured)
number related: 99% (extrapolated)

target concentration: 53 mg/m³ air
computed concentration: 47.5 mg/m³ air
MMAD: 3.04 µm; GSD: 2.21; NMAD: 0.458 µm; SMAD: 1.62 µm
respirability (% <= 5 µm): mass related: 74% (measured)
number related: 100% (extrapolated)

target concentration: 240 mg/m³ air
computed concentration: 209.2 mg/m³ air
MMAD: 5.58 µm; GSD: 1.54; NMAD: 3.15 µm; SMAD: 4.61 µm
respirability (% <= 5 µm): mass related: 41% (measured)
number related: 86% (extrapolated)

target concentration: 2386 mg/m³ air
computed concentration: 6240.6 mg/m³ air
MMAD: 32.1 µm; GSD: 2.11; NMAD: 5.97 µm; SMAD: 18.3 µm
respirability (% <= 5 µm): mass related: 1% (measured)
number related: 41% (extrapolated)
No. of animals per sex per dose:
5
10 controls
Control animals:
yes
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
- clinical signs: Day 1: multiple times
Days 2 to 14: twice daily
- body weight: pre-treatment, Day 3, Day 7, weekly thereafter
- Necropsy of survivors performed: yes
- other: Irwin screen, rectal temperature
Statistics:
means , standard deviations, statistical significance (p<=0.05 and 0.01)

Necropsy: Respiratory tract findings: RxC Chi-Square test, pairwise Fisher's test
Body weight: means , standard deviations; analysis of variances; Box-test for homogeneity; Games and Howell modification of Tukey-Kramer tesst
Particle analysis: MMAD, GSD (probit-transformed mass-related cumulative frequency distribution, logarithmized effective cut-off diameter
Concentration analytics: means, standard deviations
LC50: Maximum-Likelihood method

Results and discussion

Effect levelsopen allclose all
Key result
Sex:
male/female
Dose descriptor:
LC50
Effect level:
53 mg/L air (analytical)
Based on:
act. ingr.
Exp. duration:
4 h
Sex:
male/female
Dose descriptor:
LC50
Effect level:
0.053 mg/L air (analytical)
Based on:
act. ingr.
Exp. duration:
4 h
Mortality:
Conc. [mg/m³] deaths (M/F)
8 0/0
53 2/4
240 5/5
2386 5/5
Clinical signs:
dose dependent clinical signs
all dose-groups: hypoactivity, piloerection, unkempt appearance, labored respiration, bradypnoea

in addition at
53 mg/m³: agonal tonic convultions - males
240 mg/m³: agonal ataxia - all
agonal: apnoea, prone position, coma - males
2386 mg/m³: all animals died during eyposure
Body weight:
transient decrease in surviving animals
Gross pathology:
intermittent deaths: hepatization of lung; lung edema, hydrothorax; pale liver with accentuated lobular pattern; pale mottled kidneys with reddened pelvic area; dark spleen; reddened glandular stomach with ulcerous foci; small intestines reddened, reddish content; gastro-intestinal tract with yellow mucoid content; test-item related discolouration of inner organs.

terminal sacrifices: no effects

Applicant's summary and conclusion

Interpretation of results:
Category 2 based on GHS criteria
Conclusions:
Basic Violet 16 showed inhalation toxicity in the rat. This effect was mainly triggered by strong irritatng effects to the lung. There were signs of a concentration-dependent severe irritation of the respiratory tract as well as unspecific clinical signs and a transient decrease in body weight in all surviving animals.
The approximative LC50 for rats is 53 mg/m³ air