Reactive Orange 16

Acute Toxicity: inhalation

Administrative data

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

since September 22,1994 to October 06,1994

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Not GLP compliance but good described.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1994

Report date:

1994

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Deviations:

no

GLP compliance:

no

Test type:

fixed concentration procedure

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of disodium 6-acetamido-4-hydroxy-3-[(4-{[2-(sulfonatooxy)ethyl]sulfonyl}phenyl)diazenyl]naphthalene-2-sulfonate and sodium 6-acetamido-4-hydroxy-3-{[4(vinylsulfonyl)phenyl]diazenyl}naphthalene-2-sulfonate

Molecular formula:

n.a.

IUPAC Name:

Reaction mass of disodium 6-acetamido-4-hydroxy-3-[(4-{[2-(sulfonatooxy)ethyl]sulfonyl}phenyl)diazenyl]naphthalene-2-sulfonate and sodium 6-acetamido-4-hydroxy-3-{[4(vinylsulfonyl)phenyl]diazenyl}naphthalene-2-sulfonate

Test material form:

solid: particulate/powder

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Velaz (Prague)
- Age at study initiation: 8 weeks
- Weight at study initiation:male 176.4 +/- 14 g and female 160.8 +/- 6.35
- Fasting period before study: NO
- Housing:Plastic cage T4 (550 x 320 x 180 mm Velaz Praga)
- Diet :Fed with a mixture granulated food ALTROMIN 1320 (Velaz Prague) in a dose of 12 grams per day
- Water : ad libitum
- Acclimation period:

ENVIRONMENTAL CONDITIONS
- Temperature (°C):22 ± 3 °c
- Humidity (%): 40 - 60 %
- Photoperiod : 12 h cycle dark/light

Administration / exposure

Route of administration:

inhalation: dust

Type of inhalation exposure:

head only

Vehicle:

air

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: wright Dust Feed Unit MK2, GA 4170, L. Adams LTD., London
- Method of holding animals in test chamber:The animals were fixed in glass tubes with a diameter of 60 mm so that the face of interfering with the glass cylinder , a vertical axis,- an average of 250 mm, where there has been a dynamic linear rate changing atmosphere of 10 L / min.
- Source and rate of air: 0.6 m^3 /h
- Method of particle size determination: mycroscope
- Source and rate of air: dynamic hair system
- System of generating particulates/aerosols:through the spray unit so that the atmosphere at 'inside the inhalation chamber was homogeneous.
- Temperature, humidity, pressure in air chamber:

TEST ATMOSPHERE
- Brief description of analytical method used: Compressed air was drawn from a distribution center and Synthesia air flow was measured continuously during exposure time. The unit load was maintained 10 mm water column (pressure). Temperature and humidity were measured continuosly
- Samples taken from breathing zone: yes

Analytical verification of test atmosphere concentrations:

no

Duration of exposure:

ca. 4 h

Concentrations:

measuerd concentration: 4.85 mg/L L Air

No. of animals per sex per dose:

5 x sex x dose

Control animals:

yes

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: 2 per day
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, water and food consumption

Results and discussion

Effect levels

Effect levels 1

Sex:

male/female

Dose descriptor:

LC0

Effect level:

>= 4.85 mg/L air (nominal)

Based on:

test mat.

Exp. duration:

4 h

Effect levels 2

Sex:

male/female

Dose descriptor:

LC0

Effect level:

ca. 3.93 mg/L air

Based on:

act. ingr.

Exp. duration:

4 h

Mortality:

no mortality observed

Clinical signs:

other: other:

Body weight:

no change observed

Other findings:

Immediately after the 14 days after the observation period the animals were killed by cervical spine, disruption spinal cord and blood vessels of the neck. After an autopsy was performed, with particular attention to the comprehensive examination of the body surface and orifices, and macroscopic examination of the chest, abdominal organs and trachea, lungs, heart , liver, spleen and kidneys.
In this examination is done for all males and two females.

Any other information on results incl. tables

Immediately after application:

the animals were removed from the tubes, placed in cage and observed.
In all cases, it was noted bristly hair on the head and front legs of the sample spots.The nasal cavities and eyes were visible orange tears . Breath was regular, the number of breaths per minute 100 -110. Physical and mental mental disorders were founded . the animals werewashedand giventhem food.
60 minutes after application:
Were observed in the same clinical signs of intoxication,as inprevious examinations.
120 minutes after application:
Were observed the same clinical symptoms as in the previous examination.
2 daysafter application:
In addition to hair contact with the substance tested, in different parts of the body, they had disturbances in health.
3 to 14 days after application:
At this time there are no clinical symptoms

Discussion:

Before the test was measured by the size of microscopic particles of the sample Ostazinová V3R orange. Most particles are ranged from 2 to 4 µm.The particle size is a limiting factor to save inhaled substances in different parts of the respiratory system.
In general, the biological activity of the deposition in the regions of the respiratory tract depends on the physical propertiesof the particles.The chemical nature of these particles is of secondary importance. Their size, weight, shape, texture and solubility are limiting factors. Larger and heavier particles easily adhere to the mucos of the upper respiratory tract (HCD). In the nasal cavity, the mucosal lining of the trachea and bronchiis greater. These trapped particles are expelled from the mucos of the mouth after swallowing, and can be absorbed in the digestive tract. The deepest parts of the lung (respiratory bronchioles, alve wolari bifurcations) are virtually marked. It is these particles, although only asmall part of them to penetrate these areas, they are responsible for the birth and development of lung diseases caused by particles (Varheitetal.,1990).

Applicant's summary and conclusion

Interpretation of results:

practically nontoxic

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The substance Reactive Orange 16 is practically non toxic for a short time exposure. It shows a LD0= 4.85 mg/L (3.93 mg/L active ingredient)

Executive summary:

The sample Reactive Orange 16 was tested for acute inhalation toxicity limit. The test was performed according to OECD method No. 403. Experimental animals (10 rats Vistar strain) were exposed to inhalation of powder spray test substance for 4 hours of inhalation chamber. The concentration of test substance was measured by the gravimetric method. Prior exposure was measured microscopically initial size of the particles of the sample. The size of most particles ranged from 2 to 4 microns. Particle size is one of the limiting factors in the deposition of inhaled substances in different areas of the respiratory system. During exposure or during the period of 14 days of observation, there was no mortality of test animals. Clinic health disturbance of the animals tested respondents can be described as the result of the effect mechanically irritating dust spray on exposed mucous membranes and upper respiratory tract (HCD). Pathological-anatomical examination found in the lungs of three females, indicative of lung deposit of particles. tested substances with the subsequent development of obstructive atelectasis.