Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was conducted according to OECD TG 425, EPA TG OPPTS 870.1100 and in accordance with the Principles of Good Laboratory Practice (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:
2013
Report date:
2013

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 425 (Acute Oral Toxicity: Up-and-Down Procedure)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Deviations:
no
Principles of method if other than guideline:
not applicable
GLP compliance:
yes
Test type:
up-and-down procedure
Limit test:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
2-butoxyethyl benzoate
EC Number:
226-685-8
EC Name:
2-butoxyethyl benzoate
Cas Number:
5451-76-3
Molecular formula:
C13H18O3
IUPAC Name:
2-butoxyethyl benzoate
Test material form:
other: variable coloured liquid
Details on test material:
- Name of test material (as cited in study report): Butyl Cellosolve™ Benzoate
- Physical state: variable coloured liquid
- Analytical purity: 99.86%
- Lot/batch No.: 02112012-JLT
- Expiration date of the lot/batch: 11 February 2013
- Storage condition of test material: Ambient (+18 to +36 ºC)

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: inhouse bred (outbred)
- Age at study initiation: 8-2 weeks
- Weight at study initiation: 197.4 - 208.5 g
- Fasting period before study: overnight fasting
- Housing: Rats were housed individually in standard polysulfone cages (Size: approximately L 425 x B 266 x H 175 mm), with stainless steel top grills having facilities for pelletted food and drinking water.
- Diet (ad libitum): Ssniff® rats/mice pellet food-maintenance meal manufactured by Ssniff Spezialdiäten GmbH., Ferdinand-Gabriel-Weg 16, D-59494 SÖest, Germany, was provided to the animals.
- Water (ad libitum): Deep bore-well water passed through activated charcoal filter and exposed to UV rays in Aquaguard on-line water filter-cum-purifier manufactured by Eureka Forbes Ltd., Mumbai 400 001, India, was provided to animals in polycarbonate bottles with stainless steel sipper tubes. The water bottles were replenished once daily and the water bottles were changed once a week.
- Acclimation period: five to twenty six days under standard laboratory conditions

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-22°C
- Humidity (%): 56 – 66%
- 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:
Undiluted test substance was administered as such.
Doses:
Single animals were dosed in sequence. Each animal was dosed by oral gavage using a stainless steel ball-tipped gavage needle attached to a syringe.
Steps 1-8: The test substance was administered at the dose of 2000 mg/kg (1.97 mL/kg), 175 mg/kg (0.17 mL/kg), 550 mg/kg (0.54 mL/kg), 2000 mg/kg
(1.97 mL/kg), 550 mg/kg (0.54 mL/kg), 2000 mg/kg (1.97 mL/kg), 550 mg/kg (0.54 mL/kg) and 2000 mg/kg (1.97 mL/kg) body weight for Steps 1- 8, respectively, as a single oral gavage to the rats fasted for 16 to 17 hours (access to water was not interrupted). The animals were fed immediately after dosing.
No. of animals per sex per dose:
single
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, 1, 2, 3 and 4 hours after dose administration and once daily during days 2 – 15. Individual body weights of animals were recorded on test day 1 (pre-administration), day 8 (7 days post administration) and day 15 (14 days post administration) or at death.
- Necropsy of survivors performed: yes
Statistics:
Acute Oral Toxicity Software Program; AOT425StatPgm; AOT 425StatPgm Program User’s Manual; and Simulation Results for the AOT425StatPgm Program

Results and discussion

Preliminary study:
not applicable
Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
939.8 mg/kg bw
Based on:
test mat.
95% CL:
550 - 2 000
Remarks on result:
other: Based on the present study results, the estimated acute oral LD50 of 2-butoxyethyl benzoate is 939.8 mg/kg body weight with a 95% confidence interval of 550 to 2000 mg/kg body weight in female Wistar rats.
Mortality:
Step 1: One female rat was treated with the test substance at a limit dose of 2000 mg/kg (1.97 mL/kg) body weight. Slight piloerection, lethargy and/or red
discoloration of urine were observed in the rat on days 1 to 5 and it died on day 6.
Step 2: The second female rat was treated with the test substance at the dose of 175 mg/kg (0.17 mL/kg) body weight. There were no clinical signs of toxicity or mortality.
Step 3: The third female rat was treated with the test substance at the dose of 550 mg/kg (0.54 mL/kg) body weight. There were no clinical signs of toxicity or
mortality.
Step 4: The fourth female rat was treated with the test substance at the dose of 2000 mg/kg (1.97 mL/kg) body weight. Moderate piloerection, lethargy and/or red discoloration of urine were observed in the rat on days 2 to 4 and it died on day 4.
Step 5: The fifth female rat was treated with the test substance at the dose of 550 mg/kg (0.54 mL/kg) body weight. There were no clinical signs of toxicity or
mortality.
Step 6: The sixth female rat was treated with the test substance at the dose of 2000 mg/kg (1.97 mL/kg) body weight. Slight piloerection, lethargy and/or red
discoloration of urine were observed in the rat on days 1 to 3 and it died on day 3.
Step 7: The seventh female rat was treated with the test substance at the dose of 550 mg/kg (0.54 mL/kg) body weight. There were no clinical signs of toxicity or mortality.
Step 8: The eighth female rat was treated with the test substance at the dose of 2000 mg/kg (1.97 mL/kg) body weight. Slight piloerection, lethargy and/or red
discoloration of urine were observed in the rat on days 1 to 3 and it died on day 4.
Clinical signs:
Step 1: One female rat was treated with the test substance at a limit dose of 2000 mg/kg (1.97 mL/kg) body weight. Slight piloerection, lethargy and/or red
discoloration of urine were observed in the rat on days 1 to 5 and it died on day 6.
Step 2: The second female rat was treated with the test substance at the dose of 175 mg/kg (0.17 mL/kg) body weight. There were no clinical signs of toxicity or mortality.
Step 3: The third female rat was treated with the test substance at the dose of 550 mg/kg (0.54 mL/kg) body weight. There were no clinical signs of toxicity or
mortality.
Step 4: The fourth female rat was treated with the test substance at the dose of 2000 mg/kg (1.97 mL/kg) body weight. Moderate piloerection, lethargy and/or red discoloration of urine were observed in the rat on days 2 to 4 and it died on day 4.
Step 5: The fifth female rat was treated with the test substance at the dose of 550 mg/kg (0.54 mL/kg) body weight. There were no clinical signs of toxicity or
mortality.
Step 6: The sixth female rat was treated with the test substance at the dose of 2000 mg/kg (1.97 mL/kg) body weight. Slight piloerection, lethargy and/or red
discoloration of urine were observed in the rat on days 1 to 3 and it died on day 3.
Step 7: The seventh female rat was treated with the test substance at the dose of 550 mg/kg (0.54 mL/kg) body weight. There were no clinical signs of toxicity or mortality.
Step 8: The eighth female rat was treated with the test substance at the dose of 2000 mg/kg (1.97 mL/kg) body weight. Slight piloerection, lethargy and/or red
discoloration of urine were observed in the rat on days 1 to 3 and it died on day 4.
Body weight:
Step 1 [2000 mg/kg (1.97 mL/kg) body weight] : The rat had lost weight at death when compared to its initial weighing.
Step 2 [175 mg/kg (0.17 mL/kg) body weight] : The rat gained weight throughout the observation period.
Step 3 [550 mg/kg (0.54 mL/kg) body weight] : The rat gained weight throughout the observation period.
Step 4 [2000 mg/kg (1.97 mL/kg) body weight] : The rat had lost weight at death when compared to its initial weighing.
Step 5 [550 mg/kg (0.54 mL/kg) body weight]: The rat gained weight throughout the observation period.
Step 6 [2000 mg/kg (1.97 mL/kg) body weight] : The rat had lost weight at death when compared to its initial weighing.
Step 7 [550 mg/kg (0.54 mL/kg) body weight] : The rat gained weight throughout the observation period.
Step 8 [2000 mg/kg (1.97 mL/kg) body weight] : The rat had lost weight at death when compared to its initial weighing
Gross pathology:
Step 1 [2000 mg/kg (1.97 mL/kg) body weight] : Kidney – discolored red, multifocal; urinary bladder – distended with red colour content; stomach glandular
mucosa erosion, discolored red multifocal were observed at necropsy.
Step 2 [175 mg/kg (0.17 mL/kg) body weight] : There were no abnormalities detected at necropsy.
Step 3 [550 mg/kg (0.54 mL/kg) body weight] : There were no abnormalities detected at necropsy.
Step 4 [2000 mg/kg (1.97 mL/kg) body weight] : Liver pale; kidneys - discoloration, red multifocal; urinary bladder – distended with red colour content;
stomach - gladular mucosa - erosions, discoloration red multifocal were observed at necropsy.
Step 5 [550 mg/kg (0.54 mL/kg) body weight]: There were no abnormalities detected at necropsy.
Step 6 [2000 mg/kg (1.97 mL/kg) body weight] : Liver pale, all lobes; kidneys - discoloration red, multifocal; urinary bladder – distended with red content; stomach glandular mucosa - erosions; stomach non glandular mucosa, discoloration red multifocal were observed at necropsy.
Step 7 [550 mg/kg (0.54 mL/kg) body weight] : There were no abnormalities detected at necropsy
Step 8 [2000 mg/kg (1.97 mL/kg) body weight] : Kidneys discoloration, red multifocal and urinary bladder distended with red colour content; stomach glandular mucosa erosion, discoloration red multifocal were observed at necropsy.
Other findings:
None

Any other information on results incl. tables

None

Applicant's summary and conclusion

Interpretation of results:
Toxicity Category IV
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Based on the present study results, the estimated acute oral LD50 of 2-butoxyethyl benzoate was 939.8 mg/kg body weight with a 95% confidence interval of 550 to 2000 mg/kg body weight in female Wistar rats. According to Guidance to regulation (EC) No. 1272/2008 on classification, labelling and packaging (CLP) of substances and mixtures, 2-butoxyethyl benzoate will be classified under Acute toxicty, oral, Category 4.
Executive summary:

An acute oral toxicity test (Up-and-Down Procedure) was conducted with female Wistar rats to determine the potential of 2-butoxyethyl benzoate to produce toxicity from a single dose via the oral route.

The study was initiated with a limit test at 2000 mg/kg (step 1) with one female rat. The undiluted test substance was administered at 2000 mg/kg body weight as a single oral dose to fasted (16 - 17 hours) rats. The animal exhibited clinical signs of toxicity such as slight piloerection, lethargy and red discoloration of urine. The animal died on day 6. As per the AOT 425 statpgm, the main test was then conducted with the dose of 175 mg/kg (step 2). The rat survived with no clinical signs of toxicity, hence the next dose of 550

mg/kg (step 3) was administered to one female rat. There were no clinical signs of toxicity and the rat survived. The next dose of 2000 mg/kg (limit dose) (step 4) was then administered to one female rat. The animal exhibited clinical signs of toxicity such as moderate piloerection, lethargy and red discoloration of urine. The animal died on day 4. Hence the dosing was continued with 550 mg/kg (steps 5 and 7) and 2000 mg/kg (steps 6 and 8). There were no clinical signs of toxicity at 550 mg/kg (steps 5 and 7).

Slight piloerection, lethargy and red discoloration of urine were observed at 2000 mg/kg (steps 6 and 8) and the rats died on day 3 and day 4, respectively. The surviving rats were observed for 14 days post treatment. All the surviving rats gained body weight during the 14- day observation period. The preterminally dead rats lost weight when compared to their initial body weight.

All surviving animals were sacrificed on the scheduled Day 15 and subjected to a gross necropsy. There were no gross abnormalities detected in any of the rats at necropsy.

Liver pale; kidneys - discoloration, red multifocal; urinary bladder – distended with red colour content; stomach - glandular mucosa - erosions, discoloration red multifocal and/or stomach non glandular mucosa, discoloration red multifocal were observed in all pre-terminally dead rats at necropsy. The LD50 was determined after the 14-day observation period using the AOT 425 statpgm.

Based on the present study results, the estimated acute oral LD50 of 2-butoxyethyl benzoate was 939.8 mg/kg body weight with a 95% confidence interval of 550 to 2000 mg/kg body weight in female Wistar rats.According to Guidance to regulation (EC) No. 1272/2008 on classification, labelling and packaging (CLP) of substances and mixtures, 2-butoxyethyl benzoate will be classified under Acute toxicty, oral, Category 4.