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Acute Toxicity: oral

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Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
May 30, 2008 - February 20, 2009
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was conducted according to OECD TG425 and in accordance with the principles of GLP.
Cross-referenceopen allclose all
Reason / purpose for cross-reference:
reference to same study
Reason / purpose for cross-reference:
reference to other study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2009
Report date:
2009

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 425 (Acute Oral Toxicity: Up-and-Down Procedure)
Version / remarks:
U.S. EPA Health Effects Test Guidelines, OPPTS 870.1100 (2002), JMAFF 12-Nousan-8147, November 2000.
Deviations:
no
Principles of method if other than guideline:
not applicable
GLP compliance:
yes
Test type:
up-and-down procedure
Limit test:
yes

Test material

Constituent 1
Chemical structure
Reference substance name:
2-(2-phenoxyethoxy)ethanol
EC Number:
203-227-5
EC Name:
2-(2-phenoxyethoxy)ethanol
Cas Number:
104-68-7
Molecular formula:
C10H14O3
IUPAC Name:
2-(2-phenoxyethoxy)ethanol
Details on test material:
- Name of test material (as cited in study report): Diethylene glycol mono phenyl ether
- Physical state: Liquid
- Analytical purity: No Certificate of Analysis; Gas Chromatogram indicates 99.5% by area DiEPh
- Lot/batch No.: 200602920-11
- Expiration date of the lot/batch: 19-Feb-2009
- Stability under test conditions: Test substance was expected to be stable for the duration of testing.
- Storage condition of test material: Ambient (+18 to +36°C)
- Solubility in water/Miscibility : 3% wt.
- pH : 3.5 – 7.5 (3% aq. Soln)

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Toxicology Department of Safety Assessment, Advinus Therapeutics Private Limited, Bangalore 560 058, India
- Age at study initiation: 9-11 weeks
- Weight at study initiation: 174 - 187 g
- Fasting period before study: Fasted overnightprior to dosing
- Housing: Rats were housed individually in standard polysulfone cages (Size: approximately L 425 x B 266 x H 175 mm), with stainless steel top grill having facilities for pelletted food and drinking water.
- Diet (e.g. ad libitum): Ssniff rats/mice pellet food - maintenance meal - low in germs manufactured by Ssniff Spezialdiäten GmbH., Ferdinand- Gabriel-Weg 16, D-59494 SÖest, Germany, was provided to the animals.
- Water (e.g. ad libitum): Deep bore-well water passed through activated charcoal filter and exposed to UV rays in Aquaguard on-line water filter-cumpurifier manufactured by Eureka Forbes Ltd., Mumbai - 400 001, India, was provided to animals in polycarbonate bottles with stainless steel sipper tubes.
- Acclimation period: Seven, ten, thirteen, seventeen, twenty and twenty four days under laboratory conditions after physical examination for treatment steps.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 23°C
- Humidity (%): 30 - 70%
- Air changes (per hr): 12 - 15 air changes/hour
- Photoperiod (hrs dark / hrs light):12 hours light and 12 hours dark cycle

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
not applicable
Doses:
Animal 1: 5000 mg/kg
Animal 2: 2800 mg/kg
Animal 3: 1750 mg/kg
Animal 4: 2800 mg/kg
Animal 5: 5000 mg/kg
Animal 6: 5000 mg/kg
No. of animals per sex per dose:
not applicable
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: The animals were observed five times on test day 1 (day of administration) i.e. at 30 minutes and four times at hourly intervals (post administration) and once daily during days 2 – 15.
- Necropsy of survivors performed: At the decedents were subjected to gross necropsy as soon as possible after death. All surviving and moribund animals were necropsied at the end of the observation period. Surviving rats were euthanised with isoflurane anaesthesia. External surface of the body, all orifices, tissues and organs of the thoracic and abdominal cavities of all animals were examined. All gross findings were recorded.
Statistics:
Dose progression and stopping criteria was calculated using a dedicated software program (Acute Oral Toxicity (Guideline 425) Statistical Program) provided by the EPA. A Sigma of 0.2 was used to establish the starting dose and dose progression.

Results and discussion

Preliminary study:
not applicable
Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
3 526 mg/kg bw
Based on:
test mat.
Mortality:
Animal 6: 5000 mg/kg body weight (4.5 ml/kg B.wt ) - Coma on day 2 and died on day 2.
Clinical signs:
See below
Body weight:
Dead rats had lost body weight and all surviving rats had gained body weight through the observation period. in all treatment steps.
Gross pathology:
Animal 1, 3, 4 and 5: No abnormality detected at necropsy
Animal 2 and 6: At necropsy the findings were petechial haemorrhages in the gastric mucosa and intestine and the urinary bladder was filled with blood tinged urine.
Other findings:
None

Any other information on results incl. tables

CLINICAL SIGNS AND PRE-TERMINAL DEATHS

Step 1 : 5000 mg/kg body weight (4.5 ml/kg B.wt).

• Clinical signs : Recumbency and coma. No response to touch, sound and toe pressing. Onset of clinical signs at 30 minutes post treatment and persisted till 7 hours.

• The rat was considered as moribund and euthanized. There was no abnormality detected at necropsy.

Step 2 : 2800 mg/kg body weight (2.52 ml/kg B.wt)

• Clinical signs : Recumbency and coma. No response to touch, sound and toe pressing. Onset of clinical signs at 30 minutes post treatment and persisted till 4 hours. At 6 hours post treatment the animal showed signs of recovery and had regained consciousness, but it was dull and lethargic and continued to be dull, lethargic associated with piloerection till 24 hours post dosing. The rat showed symptoms of recumbency and urine dribbling with blood tinged urine and the rat died on day 3 post dosing. At necropsy, the findings were petechial haemorrhages in the gastric mucosa and intestine and the urinary bladder was filled with blood tinged urine.

Step 3 : 1750 mg/ kg body weight (1.58 ml/kg B.wt )

• Clinical signs : There were no clinical signs till day 15 (termination) and the rat was euthanized. There was no abnormality detected at necropsy.

Step 4 : 2800 mg/kg body weight (2.52 ml/kg B.wt)

• Clinical signs: There were no clinical signs till day 15 (termination) and the rat was euthanised. There was no abnormality detected at necropsy.

Step 5: 5000 mg/kg body weight (4.5 ml/kg B.wt)

• Clinical signs : Recumbency at 30 minutes post treatment, ataxia of limbs was observed from 1 hour to 4th hour post treatment. There were no clinical signs from day 2 till day 15 termination. The rat was euthanised. There was no abnormality detected at necropsy.

Step 6: 5000 mg/kg body weight (4.5 ml/kg B.wt )

• Clinical signs : Recumbency at 30 minutes to 4th hour post treatment on day one . Coma on day 2 and died on day 2. At necropsy the findings were petechial haemorrhages in the gastric mucosa and intestine and the urinary bladder filled with blood tinged urine.

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Based on the present study results, the estimated acute oral LD50 of the test item Diethylene glycol mono phenyl ether is 3526 mg/kg body weight and with a 95% PL.
Executive summary:

The test item Diethylene glycol mono phenyl ether was tested for acute oral toxicity in female Wistar rats. The test item was administered as received (undiluted) as a single oral dose to fasted (16 - 18 hours) rats. The treatment was initiated as follows, and the dose for the subsequent steps was arrived at by using the AOT425 StatPgm by using a sigma of 0.2.

Step 1: 5000 mg/kg body weight (4.5 ml/kg B.wt) was administered to one female rat. Note: The first animal was inadvertently administered a dose of 5000 mg/kg B.wt. This deviation had no impact on the study.

• Clinical signs: Recumbency and coma. No response to touch, sound and toe pressing. Onset of clinical signs at 30 minutes post treatment and persisted till 7 hours.

• The rat was considered as moribund and euthanised, no abnormality was detected at necropsy.

Step 2: 2800 mg/kg body weight (2.52 ml/kg B.wt) was administered to the second female rat.

• Clinical signs: Recumbency and coma. No response to touch, sound and toe pressing. Onset of clinical signs at 30 minutes post treatment and persisted till 4 hours. At 6 hours post treatment the animal showed signs of recovery and had regained consciousness, but it was dull and lethargic and continued to be dull, lethargic associated with piloerection till 24 hours post dosing. The rat showed symptoms of recumbency and urine dribbling with blood tinged urine and the rat died on day 3 post dosing. At necropsy the findings were petechial haemorrhages in the gastric mucosa and intestine and the urinary bladder was filled with blood tinged urine.

Step 3: 1750 mg/kg body weight (1.58 ml/kg body weight) was administered to the third female rat.

• Clinical signs: There were no clinical signs till day 15 (termination) and the rat was euthanised. There was no abnormality detected at necropsy.

Step 4: 2800 mg/kg body weight (2.52 ml/kg body weight) was administered to fourth female rat.

• Clinical signs: There were no clinical signs till day 15 (termination) and the rat was euthanised. There was no abnormality detected at necropsy.

Step 5: 5000 mg/kg body weight (4.5 ml/kg body weight) was administered to the fifth female rat.

• Clinical signs: Recumbency at 30 minutes post treatment, ataxia of limbs was observed from the 1st hour to 4th hour post treatment. There were no clinical signs from day 2 till day 15 termination. The rat was euthanised. There was no abnormality detected at necropsy.

Step 6: 5000 mg/kg body weight (4.5 ml/kg body weight) was administered to the sixth female rat.

• Clinical signs: Recumbency at 30 minutes to 4th hour post treatment on day one. Coma on day 2 and died on day 2. At necropsy the findings were petechial haemorrhages in the gastric mucosa and intestine and the urinary bladder filled with blood tinged urine.

As per the AOT425StatPgm the stopping criteria was met and the surviving rats were observed for 14 days post treatment and the LD50 arrived at after the 14 day observation period.

Based on the present study results, the estimated acute oral LD50 of the test item Diethylene glycol mono phenyl ether is 3526 mg/kg body weight and with a 95% PL. Confidence interval is 0 to greater than 20000, as per the agency developed Software Program AOT425 StatPgm.