2-isopropoxyethyl salicylate

Endpoint summary

Description of key information

Two studies are available that address the acute toxicity of Sakura Salicylate (2-Isopropoxyethyl salicylate), an acute oral and an acute dermal toxicity study. Both studies were conducted according to OECD guidelines and GLP principles (Klimisch 1 studies).

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

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

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

Experimental Animals
Eleven (11) females Sprague-Dawley strain SPF rats [Crl:CD(SD), Atsugi Breeding Center, Charles River Laboratories Japan, Inc.], were purchased at 7 weeks of age, and quarantined / acclimated to the test environment for over a week at the test facility. Healthy animals were selected after the quarantine/ acclimation period on the basis of the clinical signs and body weight and used in the study at 8 weeks of age. Individual body weight at the time of administration was in the range of 187 to 199 g. Group allocation was conducted 1 or 2 days before administration of each dose step. All animals to be used for group allocation were weighed, and animals were selected in such a way that the body weight of animals for the dose step group and group mean body weight between each dose step was comparable as far as possible. The animals were individually identified by marking with ear tags for small animals at the time of receipt. Labels, color-coded according to administration date, indicating the study number, administration route, dose level, sex, animal number, ear tag number and date of administration were attached to the cages. Animals remaining were excluded from the study on
day 5 after administration of the third dose step and were transferred to the animal care manager for effective utilization.
Note: The number of animals ordered according to the Protocol was 10 females, but the actual number of animals received was 11 females.

Animal Husbandry
The animals were housed in an animal room (Room Number: 701) which was controlled to maintain the temperature at 23 +/- 3°C, the relative humidity at 50 +/- 20%, the air ventilation at 10 to 15 times per hour and 12-hour lighting per day (07:00 to 19:00). The animals were housed
individually in bracket-type stainless-steel wire mesh cages (W 254 x D 350 x H 170 mm:Lead Engineering Co., Ltd.). Animals were allowed free access to pelleted diet CR-LPF (Miation-sterilized Oriental Yeast Co., Ltd., Lot Nos.: 130411, 130517) and tap water (Gotemba City Water, using water bottles). During the period of animal experiment, the actual temperature of the animal room was between 22 and 24°C, and the relative humidity was
between 39 and 75% . Analysis of contaminants in the feed was carried out for the lot by Eurofins Scientific Analytics and analysis of water contaminants was carried out on a quarterly basis according to the Waterworks Law by Shibaura Semtek Co., Ltd. Copies of all data obtained from the above facilities were filed after verifling that there were no effects on the study results.
Note: Relative humidity was over 70% between 15:50 and 16:30 on August 23, 2013, which deviated from the acceptable range specified by protocol (50 +/- 20%), due to blackout from lightning damage. However, it was a slight change and there were no abnormalities and no effects on the result of study.

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

The formulation received was used as the test substance without modification. Therefore, there was no preparation process.

Oral administration was selected in accordance with the toxicity study guidelines. The test substance was administered using a stomach tube once orally to rats which had been fasted overnight (approximately 18 hours) fiom the previous day. Individual dose volume was calculated on the basis of body weight on the day of administration (day 0 of administration).
Animals were allowed free access to feed again after the end of clinical observation at 4 hours after dosing.
The animals were observed for 14 days after administration.

Doses:

Since the acute oral toxicity of the test substance was expected to be low, the starting dose level was set at 2000 mg/kg . For the subsequent dose, the dose level for the second and third dose steps was set at 300 mg/kg in accordance with the test procedure in the toxicity study guideline "Acute Toxic Class Method".

No. of animals per sex per dose:

3

Control animals:

no

Details on study design:

Method of Observation, Measurement and Examination
The day of administration was counted as day 0 of administration and the following observation and measurement were conducted.

Observation of Clinical Condition
The animals were observed frequently for the first 6 hours after administration (from immediately after administration to 5 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours and 6 hours after administration), and once daily (between 8:43 and 10:28) thereafter for 14 days, for clinical signs such as abnormalities of external appearance, nutritional condition, posture, behavior and excretions.

Measurement of Body Weight
Body weight was measured on the day of administration (immediately before dosing) and on days 1,3,7 and 14 after administration (between 8:50 and 10:27).

Pathological Examination
Soon after discovery of the animals that died, and after survivors were sacrificed by exsanguinations (via the abdominal aorta) under isoflurane anesthesia at the end of the 14-day observation period, external appearance and organs/tissues in the cranial, thoracic and abdominal cavities were examined macroscopically. Organs/ tissues were not preserved since necropsy revealed no abnormalities.

Statistics:

Based on the death occurrence during the 14-day period after administration, approximate LD50 value was estimated.

For body weight, mean with standard deviation was calculated for each measurement day for each administration stage. In addition, body weight gain during the observation period from day 0 of administration to day 14 after administration was determined and mean with standard
deviation calculated in the same manner.

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 300 - < 2 000 mg/kg bw

Based on:

test mat.

Mortality:

All animals died in the first dose step at 2000 mg/kg: however, no deaths occurred in the second and third dose steps at 300 mg/kg. Thus, it was estimated that the LD50 was higher than 300 mg/kg but lower than 2000 mg/kg.

Clinical signs:

other: In the dose step at 2000 mg/kg , 1 animal died at 6 hours after administration and 2 animals died on day 1 after administration. The 2 animals that died on day 1 after administration showed decreased spontaneous movement and reddish urine at 6 hours after

Gross pathology:

In the animals that died in the dose step at 2000 mg/kg and in the surviving animals in both dose steps at 300 mg/kg, there were no abnormalities in the external appearance or organs/tissues in the cervical, thoracic or abdominal cavities in any animal.

Interpretation of results:

Category 4 based on GHS criteria

Remarks:

Migrated information

Conclusions:

On the basis of the results described above, it was estimated that the LD50 value of Isopropoxyethyl salicylate was higher than 300 mg/kg bw but lower than 2000 mg/kg bw when administered orally to rats once.

Executive summary:

The acute oral toxicity of Isopropoxyethyl salicylate was examined according to OECD guideline 423 and GLP principles in Sprague-Dawley rats (3 females per dose step). All animals died at 2000 mg/kg bw, however, no deaths occurred at 300 mg/kg bw. All animals exposed to 2000 mg/kg died 6 hours after administration or on day 1 after administration. The animals that died on day 1 after administration showed decreased spontaneous movement and reddish urine at 6 hours after administration. All animals in the 300 mg/kg bw showed reddish urine starting 6 hours after administration, but this effect disappeared by day 2 after administration and there were no abnormalities thereafter. At 300 mg/kg bw, body weight increase was slightly low on day 1 after administration and suppressed body weight gain was observed, but body weight increase was normal thereafter. There were no abnormalities in the results of necropsy of the animals that died or that survived. On the basis of the results described above, it was estimated that the oral LD50 value of Isopropoxyethyl salicylate was higher than 300 mg/kg but lower than 2000 mg/kg bw.

Based on these data, the LD50 for Isopropoxyethyl salicylate was determined to be between 300 and 2000 mg/kg bw, the substance is classified for oral toxicity in category 4 according to Regulation (EC) 1272/2008.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

300 mg/kg bw

Quality of whole database:

One reliable study available, no other data are present.

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

21 April 2015 - 22 December 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Version / remarks:

adopted on 24 February 1987

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: EPA, OPPTS Methods 870.1000 “Acute Toxicity Testing - Background”

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
Species and strain: Rat, Hsd: Sprague Dawley SD
Sex: Males and females (nulliparous and non-pregnant)
Age: 6 to 8 weeks old
Weight at order: 176 to 200 grams
Supplier: Charles River Italia S.p.A., Calco, Lecco, Italy
Breeder: Charles River Italia S.p.A., Calco, Lecco, Italy
Date of arrival: 09 April 2015
Weight range at arrival: 189 to 203 grams
Acclimatisation period: At least 5 days
Veterinary health check: During acclimatisation period

ENVIRONMENTAL CONDITIONS
Animals per cage: Up to 5 of one sex during acclimatisation; individually caged during the study
Housing: Clear polysulphone H-Temp solid bottomed cages (Tecniplast Gazzada S.a.r.l., Buguggiate, VA, Italy) measuring 59.5×38×20 cm during
acclimatisation period and 42.5×26.6×18.5 cm during the study with nesting material provided into suitable bedding bags.
Cage control: Daily inspected and changed as necessary (at least 3 times/week)
Water Drinking: water supplied to each cage via a water bottle
Water supply: Ad libitum
Diet: 4 RF 18 (Mucedola S.r.l., Via G. Galilei, 4, 20019, Settimo Milanese (MI) Italy)
Diet supply: Ad libitum throughout the study
Room lighting: Artificial (fluorescent tubes), daily light/dark cycle of 12/12 hours
Air changes: Approximately 15 to 20 air changes per hour
Temperature range: 22 °C±2 °C
Relative humidity range: 55%±15%

Type of coverage:

semiocclusive

Details on dermal exposure:

TEST SITE
- Area of exposure: Dorsal surface of the trunk
- % coverage: Approximately 10% of body surface
- Type of wrap if used: elastic adhesive bandage and gauze patch

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Yes, with cotton wool soaked with lukewarm water.
- Time after start of exposure: 24 hours

Duration of exposure:

The animals remained in contact with the test item for 24 hours.

Doses:

2000 mg/kg

No. of animals per sex per dose:

5 males and 5 females at 2000 mg/kg

Control animals:

not required

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Animals were observed for clinical signs the day of dosing: on dosing, approximately 1 hour after dosing, 2 hours after dosing and 5 hours after dosing, daily thereafter. All animals were weighed at allocation to the study (Day -1), on the day of dosing (Day 1) and on Days 8 and 15.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

no indication of skin irritation up to the relevant limit dose level

Mortality:

No mortality occurred.

Clinical signs:

other: No clinical signs were observed in male or female animals after treatment during the observation period.

Gross pathology:

No abnormalities were found at necropsy examination performed on all animals at termination of the study.

Interpretation of results:

GHS criteria not met

Conclusions:

The results of an acute dermal toxicity study performed according to OECD.EC guidelines and GLP principles indicate that SAKURA SALICYLATE has no toxic effect on the rat following dermal exposure over a 24 hour period at 2000 mg/kg bw. It is therefore concluded that the dermal LD50 is greater than 2000 mg/kg bw.

Executive summary:

The acute toxicity of SAKURA SALICYLATE was investigated following dermal administration of a single dose to the rat. A single dose of 2000 mg/kg bw was administered to a group of 5 male and 5 female animals for 24 hours. After a 14-day observation period, all animals were killed and subjected to necropsy examination. No mortality occurred and no signs of toxicity were observed in male or female animals during the observation period. The body weight changes observed during the study were within the expected range for this species and age of animals. No internal abnormalities were found at necropsy in the animals at termination of the study. No abnormalities were observed at the treated site. These results indicate that the test item, SAKURA SALICYLATE, has no toxic effect on the rat following dermal exposure over a 24 hour period at a level of 2000 mg/kg bw. Based on these data, the dermal LD50 for Isopropoxyethyl salicylate was determined to exceed 2000 mg/kg bw, the substance is not classified for dermal toxicity in category 4 according to Regulation (EC) 1272/2008.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

One reliable study available, no other data are present.

Additional information

Acute oral data:

The acute oral toxicity of Isopropoxyethyl salicylate was examined according to OECD guideline 423 and GLP principles in Sprague-Dawley rats (3 females per dose step). All animals died at 2000 mg/kg bw, however, no deaths occurred at 300 mg/kg bw. All animals exposed to 2000 mg/kg died 6 hours after administration or on day 1 after administration. The animals that died on day 1 after administration showed decreased spontaneous movement and reddish urine at 6 hours after administration. All animals in the 300 mg/kg bw showed reddish urine starting 6 hours after administration, but this effect disappeared by day 2 after administration and there were no abnormalities thereafter. At 300 mg/kg bw, body weight increase was slightly low on day 1 after administration and suppressed body weight gain was observed, but body weight increase was normal thereafter. There were no abnormalities in the results of necropsy of the animals that died or that survived. On the basis of the results described above, it was estimated that the oral LD50 value of Isopropoxyethyl salicylate was higher than 300 mg/kg but lower than 2000 mg/kg bw.

Acute dermal data:

The acute toxicity of Sakura salicylate was investigated following dermal administration of a single dose to the rat. A single dose of 2000 mg/kg bw was administered to a group of 5 male and 5 female animals for 24 hours. After a 14-day observation period, all animals were killed and subjected to necropsy examination. No mortality occurred and no signs of toxicity were observed in male or female animals during the observation period. The body weight changes observed during the study were within the expected range for this species and age of animals. No internal abnormalities were found at necropsy in the animals at termination of the study. No abnormalities were observed at the treated site. These results indicate that the test item, Sakura salicylate, has no toxic effect on the rat following dermal exposure over a 24 hour period at a level of 2000 mg/kg bw.

Justification for classification or non-classification

Based on the available data, the LD50 for Sakura Salicylate (Isopropoxyethyl salicylate) was determined to be between 300 and 2000 mg/kg bw, the substance is therefore classified for oral toxicity in category 4 according to Regulation (EC) 1272/2008. Since no adverse effects were seen after dermal exposure to 2000 mg/kg bw/day, Isopropoxyethyl salicylate is not classified for acute dermal toxicity according to Regulation (EC) 1272/2008.