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EC number: 216-223-3 | CAS number: 1530-32-1
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
Endpoint summary
Administrative data
Description of key information
Acute oral toxicity:
An acute oral toxicity dose (LD50) of the test chemical was considered based on experimental study conducted on rats, the LD50 value was considered in between 50-300 mg/kg bw in rats. Thus, comparing this range with the criteria of CLP regulation, the given test chemical can be classified in “Category 3” for acute oral toxicity.
Acute Inhalation toxicity:
An acute inhalation toxicity dose (LC50) of the test chemical was considered based on experimental study conducted on rats, the LC50 value was considered to be >5.0 mg/L (>5000 mg/m3) in rats. Thus, comparing this value with the criteria of CLP regulation, the given test chemical cannot be classified for acute inhalation toxicity.
Acute Dermal Toxicity:
An acute Dermal toxicity dose (LD50) of the test chemical was considered based on experimental study conducted on rats, the value was considered to be >2000 mg/kg bw in rats. Thus, comparing this value with the criteria of CLP regulation, the given test chemical cannot be classified for acute dermal toxicity.
Key value for chemical safety assessment
Acute toxicity: via oral route
Link to relevant study records
- Endpoint:
- acute toxicity: oral
- 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 423 (Acute Oral toxicity - Acute Toxic Class Method)
- Principles of method if other than guideline:
- According to OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
- GLP compliance:
- yes
- Test type:
- acute toxic class method
- Limit test:
- no
- Specific details on test material used for the study:
- - Name of test material: ETHYL TRI-PHENYL PHOSPHONIUM BROMIDE
- Appearance: White to off-white crystalline powder
- Substance type: Organic
- Physical state: solid
- AI Content (purity): 99.7%
- Storage conditions : Room temperature (20 - 30 °C)
- Batch number: ETPB1/B/14016
- Manufactured date: February, 2014
- Expiry Date : January, 2015 (Retest date)
- Other: Handling and Disposal
- Safety precautions:Aprons, masks, caps, gloves and goggles were used to ensure the health and safety of the Personnel. - Species:
- rat
- Strain:
- Wistar
- Sex:
- female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: In-House Bred
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 8- 11 weeks at the time of dosing. Healthy young adult animals were used for the study.
- Weight at study initiation: Minimum: 136 g Maximum: 155 g (Individual body weights were within ± 4% prior to treatment after overnight fasting)
- Fasting period before study: The animals were fasted for minimum 16-18 hours prior to dosing and for 4 hours post dosing, with food withheld but drinking water provided ad libitum.
- Housing:The animals were housed individually in polycarbonate cages.
- Bedding:All cages were provided with corn cobs.
- Room Sanitation:The experimental room floor and work tops were swept and mopped with disinfectant solution every day.
- Cages and water bottle:All the cages and water bottles were changed at least twice every week.
- Diet (e.g. ad libitum): All animals were provided conventional laboratory rodent diet, ad libitum
- Water (e.g. ad libitum):Aqua guard filtered tap water was provided ad libitum via drinking bottles.
- Acclimation period:Animal nos. 1-3 were acclimatized for 7 days and 4-6 for 9 days, 7-9 for 10 days and 10-12 for 6 days prior to administration of the test item.
- Identification:The animals were marked temporarily on tail, permanently on toe pad micro tattooing and cage cards. Individual cage cards were labelled with study no., study type, test system, group, dose, sex, animal number, experimental start and completion date.
ENVIRONMENTAL CONDITIONS
- Temperature (°C): Minimum: 19.60°C Maximum: 23.30°C
- Humidity (%): Minimum: 51.60 % Maximum: 64.60 %
- Air changes (per hr): More than 12 changes per hour
- Photoperiod (hrs dark / hrs light): 12:12
IN-LIFE DATES: From: April 26, 2014 To: May 26, 2014 - Route of administration:
- oral: unspecified
- Vehicle:
- corn oil
- Details on oral exposure:
- VEHICLE
- Concentration in vehicle: 2000 mg/kg, 300 mg/kg and 50 mg/kg
- Amount of vehicle (if gavage):10 ml/kg body weight.
- Justification for choice of vehicle: Corn oil was selected as a vehicle because test item was not soluble in the distilled water during solubility testing. - Doses:
- 2000 mg/kg bw, 300 mg/kg bw and 50 mg/kg bw
- No. of animals per sex per dose:
- 2000 mg/kg bw-3 female rats
300 mg/kg bw-3 female rats
50 mg/kg bw-6 female rats - Control animals:
- not specified
- Details on study design:
- - Duration of observation period following administration: 14 days
- Frequency of observations and weighing: daily
- Necropsy of survivors performed: yes, at the end of 14 day observation period, all the surviving rats were euthanised by overdose of CO2. All the animals were subjected for external and internal observations.
- Other examinations performed: Clinical Observation - After test item administration, individual animals were frequently observed at 30 minutes, 1, 2, 3 and 4 hours post dosing on day 0 (day of dosing). Subsequently, all the surviving animals were observed once a day during the 14 day observation period.
Mortality - All surviving animals were observed twice daily (morning and evening) for morbidity and mortality, throughout the acclimatization and study period.
Body weight - All rats were weighed on days 0 (prior to dosing), 7 and 14. Animals were weighed immediately after found dead. - Statistics:
- not specified
- Preliminary study:
- not specified
- Sex:
- female
- Dose descriptor:
- LD50
- Effect level:
- > 50 - < 300 mg/kg bw
- Based on:
- test mat.
- Mortality:
- At 2000 and 300 mg/ kg, all the animals were found dead on day 0 post dosing. No mortality was observed in the animals treated with 50 mg/kg dose throughtout the 14 days observation period.
- Clinical signs:
- other: At 2000 mg/kg, animal no. 1 was observed with salivation, mild tremors, mild abdominal breathing, sternal recumbency and found dead at 30 minutes post dosing. Animal no. 2 was observed with salivation, mild tremors, mild abdominal breathing, lateral recum
- Gross pathology:
- At 2000 mg/kg, all three animals were observed with wet area around mouth during external gross pathological examination.
At 300 mg/kg, animal nos. 4 and 5 were observed with no abnormality and animal no. 6 with red area around nose.
At 50 mg/kg, all the six animals were observed with no abnormality.
At 2000 mg/kg, all the animals were observed with moderate red discolouration of all lobes of the lungs, test item in stomach and mild dark colored liver during internal gross pathological examination.
At 300 mg/kg, Animal no. 4 was observed with mild red discolouration of all lobes of the lungs, test item in stomach and mild congestion of Intestine. Animal no. 5 was observed with mild red discolouration of all lobes of the lungs and test item in stomach. Animal no. 6 was observed severe hemorrhages and congestion of lungs, test item in stomach and mild congestion of Intestine.
At 50 mg/kg, all the six animals were observed with no abnormality. - Other findings:
- not specified
- Interpretation of results:
- other: Category '3' as per CLP criteria of classification and labeling.
- Conclusions:
- The acute oral LD50 value of test chemical was considered in between 50-300 mg/kg body weight. Thus by considering the CLP criteria for acute toxicity rating for the chemicals, it infers that the given test chemical exhibits acute oral toxicity in “Category 3” LD50 >50 to ≤300 mg/kg body weight.
- Executive summary:
Acute oral toxicity study of the given test chemical was conducted as per OECD No. 423 in rats. Twelve female Wistar rats were selected for acute oral toxicity study. The animals were fasted for minimum 16-18 hours prior to dosing and for 4 hours post dosing, with food withheld but drinking water provided ad libitum. The time intervals between dosing were determined by the onset, duration and severity of toxic signs. Three rats of group G1 were dosed with starting dose of 2000 mg/kg body weight and all the animals died post dosing on day 0. At 2000 mg/kg, animal no. 1 was observed with salivation, mild tremors, mild abdominal breathing, sternal recumbency and found dead at 30 minutes post dosing. Animal no. 2 was observed with salivation, mild tremors, mild abdominal breathing, lateral recumbency and found dead at 30 minutes post dosing. Animal no. 3 was observed with salivation, moderate tremors, mild abdominal breathing, lateral recumbency and found dead at 30 minutes post dosing. So, three rats of group G2 were dosed with 300 mg/kg weight and all animals died on day 0 post dosing. At 300 mg/kg, animal nos. 4 and 5 were observed with moderate lethargy at 30 minutes, mild tremors, mild abdominal breathing, lateral recumbency and found dead at 1 hour post dosing. Animal no. 6 was observed with normal sign at 30 minutes, mild lethargy at 1 and 2 hours, epistaxis at 1, 2 and 3 hours, lateral recumbency and mild abdominal breathing at 3 hours and found dead at 4 hours post dosing. So, three rats of G3 were dosed with 50 mg/kg body weight and no mortality was observed. So, another three animals of the same group G3 were dosed with 50 mg/kg body weight and no mortality was observed. Hence, further dosing was stopped. Body weights of surviving animals were recorded on day 0 (prior to dosing) 7 and 14. Mean body weight of animals treated with 50 mg/kg body weight was observed with gain on day 7 and 14, as compared to day 0. At 50 mg/kg, animal no. 7 was observed with mild diarrhoea at 3 and 4 hours and mild lethargy at 4 hours, animal no. 8 with mild epistaxis and diarrhoea at 3 and 4 hours, animal nos. 9, 10 and 11 with mild diarrhoea at 3 and 4 hours and normal rest of the observation period. Animal no. 12 was observed normal throughout the experiment period. At 2000 mg/kg, all three animals were observed with wet area around mouth during external gross pathological examination. At 300 mg/kg, animal nos. 4 and 5 were observed with no abnormality and animal no. 6 with red area around nose. At 50 mg/kg, all the six animals were observed with no abnormality. At 2000 mg/kg, all the animals were observed with moderate red discolouration of all lobes of the lungs, test item in stomach and mild dark colored liver during internal gross pathological examination. At 300 mg/kg, Animal no. 4 was observed with mild red discolouration of all lobes of the lungs, test item in stomach and mild congestion of Intestine. Animal no. 5 was observed with mild red discolouration of all lobes of the lungs and test item in stomach. Animal no. 6 was observed severe haemorrhages and congestion of lungs, test item in stomach and mild congestion of Intestine. At 50 mg/kg, all the six animals were observed with no abnormality. Under the conditions of this; the acute oral LD50 value of test chemical was considered in between 50-300 mg/kg body weight. Thus by considering the CLP criteria for acute toxicity rating for the chemicals, it infers that the given test chemical exhibits acute oral toxicity in “Category 3” LD50 >50 to ≤300 mg/kg body weight.
Reference
Table 1: Individual Animal Body Weight (g) andBody Weight Changes(%)
Sex:Female
Animal No. |
Group/ Dose (mg/kg) |
Body Weight (gram) |
Body Weight Change (%) |
||||
Day 0 |
Day 7 |
Day 14 |
Found dead |
Day 0-7 |
Day 0-14 |
||
1 |
G1/ 2000 |
146 |
- |
- |
147 |
- |
- |
2 |
143 |
- |
- |
147 |
- |
- |
|
3 |
142 |
- |
- |
143 |
- |
- |
|
4 |
G2/ 300 |
155 |
- |
- |
153 |
- |
- |
5 |
153 |
- |
- |
154 |
- |
- |
|
6 |
150 |
- |
- |
146 |
- |
- |
|
7 |
G3/ 50 |
152 |
189 |
202 |
- |
24.34 |
26.46 |
8 |
146 |
185 |
197 |
- |
26.71 |
27.57 |
|
9 |
136 |
153 |
157 |
- |
12.50 |
13.73 |
|
10 |
148 |
182 |
201 |
- |
22.97 |
29.12 |
|
11 |
143 |
181 |
196 |
- |
26.57 |
29.28 |
|
12 |
146 |
179 |
195 |
- |
22.60 |
27.37 |
Key:- = Not applicable
Table 2: Summary of Animal Body Weight (g) and Body Weight Changes (%)
Sex:Female
Group/ Dose (mg/kg) |
Rats Body Weight (g) |
Body Weight Changes (%) |
||||
Day 0 |
Day 7 |
Day 14 |
0-7 |
0-14 |
||
G1/ 2000 |
Mean |
143.67 |
- |
- |
- |
- |
SD |
2.08 |
- |
- |
- |
- |
|
N |
3 |
- |
- |
- |
- |
|
G2/ 300 |
Mean |
152.67 |
- |
- |
- |
- |
SD |
2.52 |
- |
- |
- |
- |
|
N |
3 |
- |
- |
- |
- |
|
G3/ 50 |
Mean |
145.17 |
178.17 |
191.33 |
22.62 |
25.59 |
SD |
5.38 |
12.81 |
17.05 |
5.25 |
5.91 |
|
N |
6 |
6 |
6 |
6 |
6 |
Keys:- = Not applicable,SD = Standard Deviation, n = Number of Animals
Table 3: Individual Animal Clinical Signs and Symptoms
Sex:Female
Animal No. |
Group/ Dose (mg/kg) |
Hours (Day 0) |
||||
1/2 |
1 |
2 |
3 |
4 |
||
1 |
G1/ 2000 |
145 166+ 4+ 155 2 |
- |
- |
- |
- |
2 |
145 166+ 98 4+ 2 |
- |
- |
- |
- |
|
3 |
145 166++ 4+ 98 2 |
- |
- |
- |
- |
|
4 |
G2/ 300 |
99++ |
98 4+ 166+ 2 |
- |
- |
- |
5 |
99++ |
166+ 4+ 98 2 |
- |
- |
- |
|
6 |
1 |
99+ 64 |
99+ 64 |
98 4+ 64 |
2 |
|
7 |
G3/ 50 |
1 |
1 |
1 |
49+ |
49+ 99+ |
8 |
1 |
1 |
1 |
64+ 49+ |
64+ 49+ |
|
9 |
1 |
1 |
1 |
49+ |
49+ |
|
10 |
1 |
1 |
1 |
49+ |
49+ |
|
11 |
1 |
1 |
1 |
49+ |
49+ |
|
12 |
1 |
1 |
1 |
1 |
1 |
Keys:- = Not applicable, 1 = Normal, 2 = Dead, 4 = Abdominal breathing, 49 = Diarrhoea, 64 = Epistaxis, 98 = Lateral recumbency, 99 = Lethargy, 145 = Salivation, 155 = Sternal recumbency, 166 = Tremors,+= Mild, ++ = Moderate.
Table 3: Individual Animal Clinical Signs and Symptoms(Contd...)
Animal No. |
Group/ Dose (mg/kg) |
Days post dosing |
|||||||||||||||||||||||||||||
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
9 |
10 |
11 |
12 |
13 |
14 |
||||||||||||||||||
1 |
G1/ 2000 |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
||||||||||||||||
2 |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
|||||||||||||||||
3 |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
|||||||||||||||||
4 |
G2/ 300 |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
||||||||||||||||
5 |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
|||||||||||||||||
6 |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
|||||||||||||||||
7 |
G3/ 50 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
||||||||||||||||
8 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
|||||||||||||||||
9 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
|||||||||||||||||
10 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
|||||||||||||||||
11 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
|||||||||||||||||
12 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
Keys:- = Not applicable, 1 = Normal
Table 4: Individual Animal Mortality Record
Sex:Female
Animal No. |
Group/ Dose (mg/kg) |
Day of Observation (Day 0 to 14) |
|
Morning Observations |
Evening Observations |
||
1 |
G1/ 2000 |
Found dead on day 0 post dosing |
Not applicable |
2 |
Found dead on day 0 post dosing |
Not applicable |
|
3 |
Found dead on day 0 post dosing |
Not applicable |
|
4 |
G2/ 300 |
Found dead on day 0 post dosing |
Not applicable |
5 |
Found dead on day 0 post dosing |
Not applicable |
|
6 |
Found dead on day 0 post dosing |
Not applicable |
|
7 |
G3/ 50 |
No mortality and morbidity |
No mortality and morbidity |
8 |
No mortality and morbidity |
No mortality and morbidity |
|
9 |
No mortality and morbidity |
No mortality and morbidity |
|
10 |
No mortality and morbidity |
No mortality and morbidity |
|
11 |
No mortality and morbidity |
No mortality and morbidity |
|
12 |
No mortality and morbidity |
No mortality and morbidity |
Table 5: Gross Necropsy Observation
Sex:Female
Animal No.
|
Group/ Dose (mg/kg) |
Mode of Death |
Gross Observation |
|
External |
Internal |
|||
1 |
G1/ 2000 |
Found dead |
Wet area around mouth |
Lungs: Red discolouration, all lobes (moderate) Stomach: Test item observed Liver: Dark colored (mild) |
2 |
Found dead |
Wet area around mouth |
Lungs: Red discolouration, all lobes (moderate) Stomach: Test item observed Liver: Dark colored (mild) |
|
3 |
Found dead |
Wet area around mouth |
Lungs: Red discolouration, all lobes (moderate) Stomach: Test item observed Liver: Dark colored (mild) |
|
4 |
G2/ 300 |
Found dead |
No abnormality detected |
Lungs: Red discolouration, all lobes (mild) Stomach: Test item observed Intestine: Congestion (mild) |
5 |
Found dead |
No abnormality detected |
Lungs: Red discolouration, all lobes (mild) Stomach: Test item observed |
|
6 |
Found dead |
Red area around nose |
Lungs: Hemorrhages and congestion (severe) Stomach: Test item observed Intestine: Congestion (mild) |
|
7 |
G3/ 50 |
Terminal Sacrifice |
No abnormality detected |
No abnormality detected |
8 |
Terminal Sacrifice |
No abnormality detected |
No abnormality detected |
|
9 |
Terminal Sacrifice |
No abnormality detected |
No abnormality detected |
|
10 |
Terminal Sacrifice |
No abnormality detected |
No abnormality detected |
|
11 |
Terminal Sacrifice |
No abnormality detected |
No abnormality detected |
|
12 |
Terminal Sacrifice |
No abnormality detected |
No abnormality detected |
Endpoint conclusion
- Endpoint conclusion:
- adverse effect observed
- Dose descriptor:
- LD50
- Value:
- 150 mg/kg bw
- Quality of whole database:
- Data is Klimisch 1 and from experimental study report.
Acute toxicity: via inhalation route
Link to relevant study records
- Endpoint:
- acute toxicity: inhalation
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study without detailed documentation
- Justification for type of information:
- Data is from experimental study report.
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 403 (Acute Inhalation Toxicity)
- Principles of method if other than guideline:
- The aim of this study was to assess the toxicity potential of given test chemical after inhalation exposure in rats and an observation period of 14 days.
- GLP compliance:
- no
- Test type:
- other: Acute Inhalation Toxicity
- Limit test:
- yes
- Specific details on test material used for the study:
- - Name of test material (as cited in study report):ethyltriphenylfosfonium bromide
- Molecular formula :C20H20BrP
- Molecular weight :371.25656
- Substance type:Organic
- Physical state:powder - Species:
- rat
- Strain:
- Wistar
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Age at study initiation:7 to 9 weeks
- Weight at study initiation:200 ±20g
- Identification-By cage tag and corresponding colour body marking
- Housing:Groups of five animals of similar sex in polypropylene cages with stainless steel grill top, facilities for food and water bottle, and bedding of clean paddy husk.
- Diet (e.g. ad libitum):Pelleted feed, ad libitum
- Water (e.g. ad libitum): Community tap water passed through ‘Aqua Guard on line water filter’, was kept in glass bottles, ad-libitum
- Nutritional conditions:For feeding conventional Laboratory diets may be used with an unlimited supply of drinking water.
- Acclimation period:Twenty healthy albino rats were selected and acclimatized for standard laboratory condition for period of one week in experimental room under veterinary examination.
- Randomization:After acclimatization and veterinary examination all the selected rats randomly divided into two groups of five females and five males.
ENVIRONMENTAL CONDITIONS
- Temperature (°C):19-25 deg. C
- Humidity (%):30-60%
- Air changes (per hr):Air conditioned rooms with 10-15 air changes per hour
- Photoperiod (hrs dark / hrs light):illumination cycle set to 12 hours artificial fluorescent light and 12 hours dark. - Route of administration:
- inhalation: aerosol
- Type of inhalation exposure:
- nose only
- Vehicle:
- other: Distilled water
- Details on inhalation exposure:
- GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus:cylindrical chamber built from stainless steel and glass.
- Exposure chamber volume: 8 liters
- Method of holding animals in test chamber:For the inhalation purpose the rats were placed in polycarbonate holder tubes positioned radically around exposure chamber, so that only the snouts and nostrils of the animals were exposed to the aerosol.
- Source and rate of air: The chamber had a inner and outer chamber to minimize the fluctuation in concentration and temperature.
- Treatment of exhaust air: The exhaust air was decontaminated by subsequent passage through 1% NaOH solution, silica gel and activated charcoal filters.
- Temperature, humidity, pressure in air chamber: The chamber was maintained at a slightly negative pressure to prevent leakage of the test atmosphere from the system, as well as its dilution with outside air. - Analytical verification of test atmosphere concentrations:
- not specified
- Duration of exposure:
- 4 h
- Concentrations:
- Group I – Limit test (5 mg/L)
Group II – Confirmatory test (5 mg/L) - No. of animals per sex per dose:
- 10 (5 males and 5 females)
- Control animals:
- not specified
- Details on study design:
- - Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical Signs: The treated animals were observed for signs of intoxication, at various intervals for first six hours after dosing and thereafter twice a day for 14 days. Any clinical signs in the treated group, if observed, recorded and mentioned in the report.
Mortality: All the animals were observed for mortality at various intervals for first six hours on the day of dosing and thereafter twice a day for 14 days.
Body weight: The body weight of all the animals was observed weekly on day 0 (Before treatment), 7th and 14th (post treatment).
- Necropsy of survivors performed: yes, necropsy was carried out on all the animals that died during the study or surviving animals were sacrificed at the end of the study to observe any gross pathological changes. - Statistics:
- not specified
- Preliminary study:
- LIMIT TEST: Ten healthy Wistar albino rats of both sexes (5 male and 5 female) of body weight 200±20 gm were selected for study after acclimatization. The test group of animals was exposed to aerosol at the concentration of 5 mg/L for period of 4 hrs. After exposure all the animals were closely observed for clinical signs of toxicity at various intervals such as 1 hr, 2 hrs, 4 hrs, and 6 hrs on the day of test compound aerosol exposure and later on twice a day throughout the experimentation period of 14 days. The necropsy was performed on all the animals at termination of experiment. All the albino rats exposed to aerosol of the test compound at the concentration of 5 mg/L did not show any clinical signs of intoxication. Furthermore, no mortality was observed throughout the period of observation.Necropsy findings: The necropsy was performed on all the animals at the termination of study did not show any gross pathological changes.
- Sex:
- male/female
- Dose descriptor:
- LC50
- Effect level:
- > 5 mg/L air
- Based on:
- test mat.
- Exp. duration:
- 4 h
- Remarks on result:
- other: no mortality was observed
- Mortality:
- The test compound did not elicit any mortality at the tested dose level of 5.0 mg/L throughout the period of observation after exposure.
- Clinical signs:
- other: The test compound did not elicit any clinical signs of intoxication at the tested aerosol concentration of 5 mg/L observed for the period of 14 days.
- Body weight:
- The body weight of all the animals recorded individually on day 0th and thereafter 7th and 14th (post treatment) showed normal increase as compared to day 0th.
- Gross pathology:
- NECROPSY FINDING
EXTERNAL
i.Skin- Skin and hair coat was observed wet.
ii.All external orifices- Normal
B. INTERNAL
i. Subcutaneous- No changes were observed.
ii. Superficial and deep lymph nodes- No change in mesenteric lymph node.
ABDOMINAL CAVITY
i.Opening and general examination- In the abdominal cavity all the organs were present in normal position.
ii.Spleen- No changes were recorded.
iii.Digestive system- No gross changes were observed in stomach and intestine.
iv.Liver and biliary ducts- No gross pathological changes were observed
v.Excretory system- No gross pathological changes were observed.
vi.Adrenal- Observed normal.
vii.Male/female genital organs – Showed normal colour, consistency and no inflammatory changes.
2. THORACIC CAVITY
i.Opening and general examination- Thoracic cavity was found to be normal without any fluid, mucous or blood etc.
ii.Lungs- No changes were recorded.
iii.Heart- No changes were observed in color and consistency. Heart found normal.
iv.Thyroid- Normal in shape, size and surface.
3. CRANIAL CAVITY
Brain- Normal in shape and size. - Other findings:
- not specified
- Interpretation of results:
- other: Not classified
- Conclusions:
- Based on the results obtained from above investigation, the acute inhalation toxicity dose (LC50) was considered to be >5.0 mg/l. Hence, it can be concluded that the test compound is non toxic to male and female Wistar rats by inhalation route via aerosol.
- Executive summary:
The acute inhalation study of the given test chemical was conducted in albino rat according to OECD-Guideline -403 for testing of chemicals. In limit test, ten healthy Wistar albino rats of both sexes (5 male and 5 female) of body weight 200±20 gm were selected for study after acclimatization. The test group of animals was exposed to aerosol at the concentration of 5 mg/L for period of 4 hrs. After exposure all the animals were closely observed for clinical signs of toxicity at various intervals such as 1 hr, 2 hrs, 4 hrs, and 6 hrs on the day of test compound aerosol exposure and later on twice a day throughout the experimentation period of 14 days. The necropsy was performed on all the animals at termination of experiment. All the albino rats exposed to aerosol of the test compound at the concentration of 5 mg/L did not show any clinical signs of intoxication. Furthermore, no mortality was observed throughout the period of observation. The necropsy was performed on all the animals at the termination of study did not show any gross pathological changes. After 72 hrs, the result obtained from limit test was confirmed in another 10 animal of both sex at similar concentration following same guideline. Ten healthy Wistar albino rats of both sexes (body weight 200±20 gm) were selected for study after acclimatization. The animals of test group were exposed to aerosol of the test compound at the concentration of 5 mg/L for period of 4 hrs. After exposure all the animals were closely observed for any clinical signs of toxicity at various intervals such as 1 hr, 2 hrs, 4 hrs, and 6 hrs on the day of test compound aerosol exposure and later on twice a day throughout the experimentation period of 14 days. The necropsy was performed on all the animals at termination of experiment. All the albino rats exposed to aerosol at the concentration of 5 mg/L did not show any clinical signs of intoxication. Again there was no mortality recorded during the entire observation period. The body weight of animals exposed to test compound, observed on day 0th (pre treatment) and day 7th (post treatment) did not differ significantly as compared to day 0th. Whereas, body weight of animals observed on day 14th showed normal increase as compared to day 0th. The necropsy was performed on all the animals at the termination of study did not show any gross pathological changes. Based on the results obtained from above investigation, the acute inhalation toxicity dose (LC50) was considered to be >5.0 mg/l. Hence, it can be concluded that the test compound is non toxic to male and female Wistar rats by inhalation route via aerosol.
Reference
TABLE – 3
CLINICAL SIGNS AND MORTALITY
Group: I Dose: 5.0 mg/L
WISTAR ALBNINO RATS
Parameters |
Incidence of Clinical Signs Observed after Dosing on |
Mortality |
|||||||||||||||||||
Day 0 |
DAY |
||||||||||||||||||||
Min |
Hour |
||||||||||||||||||||
30 |
1 |
2 |
4 |
6 |
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
9 |
10 |
11 |
12 |
13 |
14 |
Total |
% |
|
Mortality (total) |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0/10 |
0 |
Clinical Signs |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
TABLE – 3 Contd…………….
CLINICAL SIGNS AND MORTALITY
Group: II Dose: 5.0 mg/L
WISTAR ALBINO RATS
Parameters |
Incidence of Clinical Signs Observed after Dosing on |
Mortality |
|||||||||||||||||||
Day 0 |
DAY |
||||||||||||||||||||
Min |
Hour |
||||||||||||||||||||
30 |
1 |
2 |
4 |
6 |
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
9 |
10 |
11 |
12 |
13 |
14 |
Total |
% |
|
Mortality (total) |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0/10 |
0 |
Clinical Signs |
0 |
0 |
0 |
0 |
6 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 = No clinical sign (Normal)
+ = Mild
++ = Moderate
+++ = High
++++ = Severe
TABLE – 4
SUMMARY OF NECROPSY FINDING
S. No. |
Fate
|
Wistar albino rats |
|
Dose (mg/l) |
|||
5.0 (limit test) |
5.0 (confirmatory test) |
||
1 |
Terminal sacrifice |
10/10 |
10/10 |
2 |
Found Dead |
0/10 |
0/10 |
3 |
Abnormalities detected |
0/10 |
0/10 |
TABLE – 5
INDIVIDUAL ANIMAL FATE & NECROPSY FINDINGS
Group: I 5.0 mg/L
WISTAR ALBINO RATS
Animal ID |
Fate |
Time |
Gross Findings |
20173-1 |
TS |
Day 14 |
NAD |
20173-2 |
TS |
Day 14 |
NAD |
20173-3 |
TS |
Day 14 |
NAD |
20173-4 |
TS |
Day 14 |
NAD |
20173-5 |
TS |
Day 14 |
NAD |
20173-6 |
TS |
Day 14 |
NAD |
20173-7 |
TS |
Day 14 |
NAD |
20173-8 |
TS |
Day 14 |
NAD |
20173-9 |
TS |
Day 14 |
NAD |
20173-10 |
TS |
Day 14 |
NAD |
Day 0 is the day of exposure
TS=Terminal Sacrifice
NAD=No Abnormality Detected
FD=Found Dead
TABLE- 5 Contd………………
INDIVIDUAL ANIMAL FATE & NECROPSY FINDINGS
Group: II 5.0 mg/L
WISTAR ALBINO RATS
Animal ID |
Fate |
Time |
Gross Findings |
20173-11 |
TS |
Day 14 |
NAD |
20173-12 |
TS |
Day 14 |
NAD |
20173-13 |
TS |
Day 14 |
NAD |
20173-14 |
TS |
Day 14 |
NAD |
20173-15 |
TS |
Day 14 |
NAD |
20173-16 |
TS |
Day 14 |
NAD |
20173-17 |
TS |
Day 14 |
NAD |
20173-18 |
TS |
Day 14 |
NAD |
20173-19 |
TS |
Day 14 |
NAD |
20173-20 |
TS |
Day 14 |
NAD |
Day 0 is the day of exposure
TS=Terminal Sacrifice
NAD=No Abnormality Detected
FD=Found Dead
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed
- Dose descriptor:
- LC50
- Value:
- 5 000 mg/m³ air
- Quality of whole database:
- Data is Klimisch 2 and from experimental study report.
Acute toxicity: via dermal route
Link to relevant study records
- Endpoint:
- acute toxicity: dermal
- 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 402 (Acute Dermal Toxicity)
- Principles of method if other than guideline:
- According to OECD Guideline 402 (Acute Dermal Toxicity)
- GLP compliance:
- yes
- Test type:
- fixed dose procedure
- Limit test:
- yes
- Specific details on test material used for the study:
- Identification : ETHYL TRI-PHENYL PHOSPHONIUM BROMIDE
Appearance : White to off-white crystalline powder
Batch number : ETPB1/B/14016
AI Content (purity) : 99.79%
Storage conditions : Room temperature (20 - 30 °C)
Manufactured date : February, 2014
Expiry Date : January, 2016 (Retest date)
Handling and Disposal
Safety precautions :Aprons, masks, caps, gloves and goggles were used to ensure the health and safety of the Personnel. - Species:
- rat
- Strain:
- Wistar
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: In-House Bred
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: Healthy young adult animals were used for the study
- Weight at study initiation: Male:Minimum: 264 g and Maximum: 298 g
(Prior to Treatment)Female:Minimum: 242 g and Maximum: 281 g
- Housing:The animals were housed individually in polycarbonate cages.
- Bedding:All cages were provided with corn cobs
- Room Sanitation:The experimental room floor and work tops were swept and mopped with disinfectant solution every day.
- Cages and water bottle:All the cages and water bottles were changed at least twice every week.
- Diet (e.g. ad libitum): All animals were provided conventional laboratory rodent diet, ad libitum.
- Water (e.g. ad libitum):Aqua guard filtered tap water was provided ad libitum via drinking bottles.
- Acclimation period:All animals were acclimatized to the test conditions for 7 days prior to administration of the test item.
- Identification:During Acclimatization, animals were temporarily marked by permanent marker, on their tails. After acclimatization, the animals were marked by toe pad micro tattooing and cage cards. Individual cage cards were labelled with study no., study type, test system, sex, animal number, experimental start and completion date.
- Randomization:Animals were selected manually. No computer generated randomization program was used.
ENVIRONMENTAL CONDITIONS
- Temperature (°C): Minimum: 19.70 °C Maximum: 23.30 °C
- Humidity (%): Minimum: 51.60% Maximum: 64.60%
- Air changes (per hr): More than 12 changes per hour
- Photoperiod (hrs dark / hrs light): 12:12
IN-LIFE DATES: From:April 26, 2014 To: May 17, 2014 - Type of coverage:
- occlusive
- Vehicle:
- other: distilled water
- Details on dermal exposure:
- TEST SITE
- Area of exposure: dorsal area of rat skin
- % coverage: approximately 10%
- Type of wrap if used: Test item was held in contact with the skin with a porous gauze dressing and non-irritating tape
REMOVAL OF TEST SUBSTANCE
- Washing (if done): At the end of the exposure period, residual test item was removed by using distilled water.
- Time after start of exposure: 24-hour
VEHICLE
- Amount(s) applied (volume or weight with unit): 0.2 ml - Duration of exposure:
- 24 hours
- Doses:
- 2000 mg/kg bw
- No. of animals per sex per dose:
- 10 (Five per sex)
- Control animals:
- not specified
- Details on study design:
- - Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical Observation - After test item administration, individual animals were frequently observed at 1, 2, 3 and 4 hours post dosing on day 0 (day of dosing). Subsequently, all animals were observed once a day during the 14 day observation period.
Mortality - Animals were observed twice daily for any mortality during the experimental period.
Body weight - All rats were weighed on days 0 (prior to dosing), 7 and 14.
Local Signs/Skin Reactions - All animals were observed once daily during days 1-14 (in common with clinical signs).
- Necropsy of survivors performed: yes, at the end of 14 day observation period, all the surviving rats were euthanised by overdose of CO2 and subjected to gross pathology examination, for external and internal observations. - Statistics:
- No statistical analysis was performed since the study was terminated with limit test.
- Preliminary study:
- Limit Test - Five male and five female wistar rats were treated with test item by a single dermal application at the dose level of 2000 mg/kg body weight. Since no test item related mortality was observed, the study was terminated with limit test only.
- Sex:
- male/female
- Dose descriptor:
- LD50
- Effect level:
- > 2 000 mg/kg bw
- Based on:
- test mat.
- Remarks on result:
- other: No mortality was observed
- Mortality:
- No mortality was observed at limit dose of 2000 mg/kg body weight of test item during the 14 day observation period.
- Clinical signs:
- other: All the animals were observed with normal clinical signs throughout the experimental period.
- Gross pathology:
- The external and internal gross pathological observation of all terminally sacrificed animals did not show any pathological abnormality.
- Other findings:
- not specified
- Interpretation of results:
- other: Not classified
- Conclusions:
- Under the conditions of this; acute dermal toxicity dose (LD50) value for given test chemical was considered to be >2000 mg/kg body weight. Thus by considering the CLP criteria for acute toxicity rating for the chemicals, it infers that the given test chemical does not exhibit acute dermal toxicity i.e it is acutely non toxic to animals.
- Executive summary:
Acute dermal toxicity study of the given test chemical was conducted as per OECD No.402 in Wistar rats. Five male and five female healthy young adult rats were randomly selected and used for conducting acute dermal toxicity study. Rats free from injury and irritation of skin were selected for the study. Twenty four hours prior to dermal application of test item, approximately 10% of body surface area of each rat was clipped. A limit dose of 2000 mg/ kg body weight of test item moistened with 0.2 ml distilled water was applied by single dermal application and observed for 14 days after treatment. On test day 0, an amount of test item (pulverized) moistened with 0.2 ml distilled water was applied directly on the intact skin of clipped area of rats; the porous gauze dressing was put on to the intact skin of clipped area. This porous gauze dressing was covered with a non-irritating tape. After the 24-hour application period, the dressings were removed and the skin was gently wiped with distilled water. The skin reactions were assessed. The animals were observed daily for mortality and clinical signs, during the acclimatization period. All animals were observed for clinical signs at approximately 1, 2, 3 and 4 hours after treatment on day 0 and once daily during test days 1‑14. Mortality was recorded after application on test day 0 and twice daily during days 1-14 (at least once on the day of sacrifice). Local signs / Skin reactions were observed daily from test days 1-14 (in common with clinical signs). Body weights were recorded on day 0 (prior to application) and on day 7 and 14. All animals were necropsied and examined macroscopically. No mortality was observed in any animal till the end of the experimental period. All the animals were observed with normal clinical signs throughout the experimental period. In males, mean body weight was observed with increase on day 7 and 14, as compared to day 0. In females, mean body weight was observed with decrease on day 7 and increase on day 14, as compared to day 0. The external and internal gross pathological observation of all terminally sacrificed animals did not show any pathological abnormality. Under the conditions of this; acute dermal toxicity dose (LD50) value for given test chemical was considered to be >2000 mg/kg body weight. Thus by considering the CLP criteria for acute toxicity rating for the chemicals, it infers that the given test chemical does not exhibit acute dermal toxicity i.e it is acutely non toxic to animals.
Reference
Table 1: Individual Animal Body Weight (g) andBody Weight Changes(%)
Dose:2000 mg/ kg bodyweight
Animal No. |
Sex |
Body Weight (gram) |
Body Weight Change (%) |
|||
Day 0 |
Day 7 |
Day 14 |
Day 0-7 |
Day 0-14 |
||
1 |
Male |
264 |
283 |
316 |
7.20 |
19.70 |
2 |
298 |
318 |
340 |
6.71 |
14.09 |
|
3 |
287 |
299 |
319 |
4.18 |
11.15 |
|
4 |
289 |
305 |
330 |
5.54 |
14.19 |
|
5 |
275 |
274 |
290 |
-0.36 |
5.45 |
|
6 |
Female |
249 |
244 |
247 |
-2.01 |
-0.80 |
7 |
274 |
272 |
282 |
-0.73 |
2.92 |
|
8 |
281 |
280 |
283 |
-0.36 |
0.71 |
|
9 |
242 |
242 |
253 |
0.00 |
4.55 |
|
10 |
256 |
255 |
255 |
-0.39 |
-0.39 |
Table 2: Individual Animal Clinical Signs and Symptoms
Dose:2000 mg/kg body weight
Animal No. |
Sex |
Hour(s) - Day 0 |
Day |
|||||||||
1 |
2 |
3 |
4 |
1 |
2 |
3 |
4 |
5 |
6 |
7 |
||
1 |
Male |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
2 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
|
3 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
|
4 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
|
5 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
|
6 |
Female |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
7 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
|
8 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
|
9 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
|
10 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
Animal No. |
Sex |
Day |
||||||
8 |
9 |
10 |
11 |
12 |
13 |
14 |
||
1 |
Male |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
2 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
|
3 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
|
4 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
|
5 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
|
6 |
Female |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
7 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
|
8 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
|
9 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
|
10 |
1 |
1 |
1 |
1 |
1 |
1 |
1 |
Keys: 1 = Normal
Table 4:Summaryof Animal Body Weight (g) and Body Weight Changes (%)
Dose:2000 mg/kg body weight
Sex |
Body Weight (gram) |
Body Weight Changes (%) |
||||
Day 0 |
Day 7 |
Day 14 |
Day 0-7 |
Day 0-14 |
||
Male |
Mean |
282.60 |
295.80 |
319.00 |
4.65 |
12.92 |
SD |
13.24 |
17.51 |
18.79 |
3.04 |
5.19 |
|
n |
5 |
5 |
5 |
5 |
5 |
|
Female |
Mean |
260.40 |
258.60 |
264.00 |
-0.70 |
1.40 |
SD |
16.56 |
16.88 |
17.15 |
0.78 |
2.28 |
|
n |
5 |
5 |
5 |
5 |
5 |
Keys:SD= Standard deviation, n = Number of animals
Table 5: GrossNecropsyObservation
Dose:2000 mg/kg body weight Mode of Death:Terminal Sacrifice
Animal No. |
Sex |
Gross Observation |
|
External |
Internal |
||
1 |
Male |
No abnormalities detected |
No abnormalities detected |
2 |
No abnormalities detected |
No abnormalities detected |
|
3 |
No abnormalities detected |
No abnormalities detected |
|
4 |
No abnormalities detected |
No abnormalities detected |
|
5 |
No abnormalities detected |
No abnormalities detected |
|
6 |
Female |
No abnormalities detected |
No abnormalities detected |
7 |
No abnormalities detected |
No abnormalities detected |
|
8 |
No abnormalities detected |
No abnormalities detected |
|
9 |
No abnormalities detected |
No abnormalities detected |
|
10 |
No abnormalities detected |
No abnormalities detected |
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed
- Dose descriptor:
- LD50
- Value:
- 2 000 mg/kg bw
- Quality of whole database:
- Data is Klimisch 1 and from experimental study report.
Additional information
Acute oral toxicity:
An acute oral toxicity study of the given test chemical was conducted as per OECD TG No. 423 in rats. Twelve female Wistar rats were selected for acute oral toxicity study. The animals were fasted for minimum 16-18 hours prior to dosing and for 4 hours post dosing, with food withheld but drinking water provided ad libitum. The time intervals between dosing were determined by the onset, duration and severity of toxic signs. Three rats of group G1 were dosed with starting dose of 2000 mg/kg body weight and all the animals died post dosing on day 0. At 2000 mg/kg, animal no. 1 was observed with salivation, mild tremors, mild abdominal breathing, sternal recumbency and found dead at 30 minutes post dosing. Animal no. 2 was observed with salivation, mild tremors, mild abdominal breathing, lateral recumbency and found dead at 30 minutes post dosing. Animal no. 3 was observed with salivation, moderate tremors, mild abdominal breathing, lateral recumbency and found dead at 30 minutes post dosing. So, three rats of group G2 were dosed with 300 mg/kg weight and all animals died on day 0 post dosing. At 300 mg/kg, animal nos. 4 and 5 were observed with moderate lethargy at 30 minutes, mild tremors, mild abdominal breathing, lateral recumbency and found dead at 1 hour post dosing. Animal no. 6 was observed with normal sign at 30 minutes, mild lethargy at 1 and 2 hours, epistaxis at 1, 2 and 3 hours, lateral recumbency and mild abdominal breathing at 3 hours and found dead at 4 hours post dosing. So, three rats of G3 were dosed with 50 mg/kg body weight and no mortality was observed. So, another three animals of the same group G3 were dosed with 50 mg/kg body weight and no mortality was observed. Hence, further dosing was stopped. Body weights of surviving animals were recorded on day 0 (prior to dosing) 7 and 14. Mean body weight of animals treated with 50 mg/kg body weight was observed with gain on day 7 and 14, as compared to day 0. At 50 mg/kg, animal no. 7 was observed with mild diarrhoea at 3 and 4 hours and mild lethargy at 4 hours, animal no. 8 with mild epistaxis and diarrhoea at 3 and 4 hours, animal nos. 9, 10 and 11 with mild diarrhoea at 3 and 4 hours and normal rest of the observation period. Animal no. 12 was observed normal throughout the experiment period. At 2000 mg/kg, all three animals were observed with wet area around mouth during external gross pathological examination. At 300 mg/kg, animal nos. 4 and 5 were observed with no abnormality and animal no. 6 with red area around nose. At 50 mg/kg, all the six animals were observed with no abnormality. At 2000 mg/kg, all the animals were observed with moderate red discolouration of all lobes of the lungs, test item in stomach and mild dark colored liver during internal gross pathological examination. At 300 mg/kg, Animal no. 4 was observed with mild red discolouration of all lobes of the lungs, test item in stomach and mild congestion of Intestine. Animal no. 5 was observed with mild red discolouration of all lobes of the lungs and test item in stomach. Animal no. 6 was observed severe haemorrhages and congestion of lungs, test item in stomach and mild congestion of Intestine. At 50 mg/kg, all the six animals were observed with no abnormality. Under the conditions of this, all rats were died at 300 mg/kg bw, hence the LD100 was considered to be 300 mg/kg bw. Considering this value, the LD50 value can be assumed to be 150 mg/kg bw. Therefore, comparing this value with the criteria of CLP regulation, the given test chemical can be classified in “Category 3 (>50 to ≤300)” for acute oral toxicity.
Acute Inhalation toxicity:
The acute inhalation study of the given test chemical was conducted in albino rat according to OECD-Guideline -403 for testing of chemicals. In limit test, ten healthy Wistar albino rats of both sexes (5 male and 5 female) of body weight 200±20 gm were selected for study after acclimatization. The test group of animals was exposed to aerosol at the concentration of 5 mg/L for period of 4 hrs. After exposure all the animals were closely observed for clinical signs of toxicity at various intervals such as 1 hr, 2 hrs, 4 hrs, and 6 hrs on the day of test compound aerosol exposure and later on twice a day throughout the experimentation period of 14 days. The necropsy was performed on all the animals at termination of experiment. All the albino rats exposed to aerosol of the test compound at the concentration of 5 mg/L did not show any clinical signs of intoxication. Furthermore, no mortality was observed throughout the period of observation. The necropsy was performed on all the animals at the termination of study did not show any gross pathological changes. After 72 hrs, the result obtained from limit test was confirmed in another 10 animal of both sex at similar concentration following same guideline. Ten healthy Wistar albino rats of both sexes (body weight 200±20 gm) were selected for study after acclimatization. The animals of test group were exposed to aerosol of the test compound at the concentration of 5 mg/L for period of 4 hrs. After exposure all the animals were closely observed for any clinical signs of toxicity at various intervals such as 1 hr, 2 hrs, 4 hrs, and 6 hrs on the day of test compound aerosol exposure and later on twice a day throughout the experimentation period of 14 days. The necropsy was performed on all the animals at termination of experiment. All the albino rats exposed to aerosol at the concentration of 5 mg/L did not show any clinical signs of intoxication. Again there was no mortality recorded during the entire observation period. The body weight of animals exposed to test compound, observed on day 0th (pre treatment) and day 7th (post treatment) did not differ significantly as compared to day 0th. Whereas, body weight of animals observed on day 14th showed normal increase as compared to day 0th. The necropsy was performed on all the animals at the termination of study did not show any gross pathological changes. Based on the results obtained from above investigation, the acute inhalation toxicity dose (LC50) was considered to be >5.0 mg/l. Hence, it can be concluded that the test compound is non toxic to male and female Wistar rats by inhalation route via aerosol.
Acute Dermal toxicity:
An acute dermal toxicity study of the given test chemical was conducted as per OECD No.402 in Wistar rats. Five male and five female healthy young adult rats were randomly selected and used for conducting acute dermal toxicity study. Rats free from injury and irritation of skin were selected for the study. Twenty four hours prior to dermal application of test item, approximately 10% of body surface area of each rat was clipped. A limit dose of 2000 mg/ kg body weight of test item moistened with 0.2 ml distilled water was applied by single dermal application and observed for 14 days after treatment. On test day 0, an amount of test item (pulverized) moistened with 0.2 ml distilled water was applied directly on the intact skin of clipped area of rats; the porous gauze dressing was put on to the intact skin of clipped area. This porous gauze dressing was covered with a non-irritating tape. After the 24-hour application period, the dressings were removed and the skin was gently wiped with distilled water. The skin reactions were assessed. The animals were observed daily for mortality and clinical signs, during the acclimatization period. All animals were observed for clinical signs at approximately 1, 2, 3 and 4 hours after treatment on day 0 and once daily during test days 1‑14. Mortality was recorded after application on test day 0 and twice daily during days 1-14 (at least once on the day of sacrifice). Local signs / Skin reactions were observed daily from test days 1-14 (in common with clinical signs). Body weights were recorded on day 0 (prior to application) and on day 7 and 14. All animals were necropsied and examined macroscopically. No mortality was observed in any animal till the end of the experimental period. All the animals were observed with normal clinical signs throughout the experimental period. In males, mean body weight was observed with increase on day 7 and 14, as compared to day 0. In females, mean body weight was observed with decrease on day 7 and increase on day 14, as compared to day 0. The external and internal gross pathological observation of all terminally sacrificed animals did not show any pathological abnormality. Under the conditions of this; acute dermal toxicity dose (LD50) value for given test chemical was considered to be >2000 mg/kg body weight. Thus by considering the CLP criteria for acute toxicity rating for the chemicals, it infers that the given test chemical does not exhibit acute dermal toxicity i.e. it is acutely non toxic to animals.
Justification for classification or non-classification
Based on the above experimental studies on the test chemical, it can be concluded that LD50 value is between 50 to 300 mg/kg bw, for acute oral toxicity; LD50 value is >2000 mg/kg bw, for acute dermal toxicity; and LC50 value is >5 mg/L, for acute inhalation toxicity. Thus, comparing these values with the criteria of CLP regulation, the given test chemical can be classified in “Category 3” for acute oral toxicity and cannot be classified for acute dermal and acute inhalation toxicity.
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