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Administrative data

Description of key information

A sample of D-lactide was examined for acute oral toxicity in an experiment according to OECD guideline 423 with female rats (limit testing). A dose level of 2000 mg/kg body weight was examined. No mortality or distinct clinical signs were observed after treatment of 6 females with the 2000 mg/kg dose level.
In an acute dermal toxicity study (limit test), a group of young adult Wistar rats (5 males and 5 females) was dermally exposed to D-lactide (purity 99%) in polyethylene glycol 400 for 24 hours to approximately 10% of body surface area at 2000 mg/kg bw. Animals were observed for 14 days. No mortality occurred. There were no treatment related clinical signs, necropsy findings or changes in body weight.
Lactic acid is used as an read-across partner for D-lactide and in an acute inhalation toxicity study in rats with lactid acid a LC50 of > 7.94 mg/L air was determined.

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
Study period:
2010-05-28 to 2010-09-27
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP study
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
other: JMAFF Notification No 8147 (2000), including the most recent partial revisions
Deviations:
no
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany.
- Age at study initiation: approximately 8 weeks old.
- Weight at study initiation:Body weight variation did not exceed ± 20 % of the sex mean (females: 159 g).
- Fasting period before study: Animal were deprived of food overnight prior to dosing and until 3-4 hours after administration of the test substances. Water was available.
- Housing: Group housing of 3 animals per cage inlabeled Macrolon cages (MIV type; height 18 cm) containing sterilized sawdust as bedding material (Litalabo, S.P.P.S., Argenteuil, France) and paper as cage-enrichment (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom).
- Diet (e.g. ad libitum): Free access to pelleted rodent diet (SM R/M-Z from SSNIFF@Spezialdiäten GmbH, Soest, Germany).
- Water (e.g. ad libitum): Free access to tap water.
- Acclimation period: At least 5 days before start of treatment under laboratory conditions.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.0 ± 3.0 °C (actual range: 19.8-21.5 °C)
- Humidity (%): Relative humidity of 40-70 % (actual range: 38-75 %)
- Air changes (per h): Approximately 15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescent light and 12 hours darkness per day

IN-LIFE DATES: From 04 June 2010 to: 23 June 2010
Route of administration:
oral: gavage
Vehicle:
polyethylene glycol
Details on oral exposure:
VEHICLE
- Dose level (volume): 2000 mg/kg (10 ml/kg) body weight.
- Justification for choice of vehicle: The vehicle was selected based on trial formulations performed at NOTOX and on test substance data supplied by the sponsor.

MAXIMUM DOSE VOLUME APPLIED: 2000 mg/kg (10 ml/kg) body weight

DOSAGE PREPARATION (if unusual): The vehicle was dehydrated before the formulations were prepared. The formulations (w/w) were prepared within 4 hours prior to dosing. Homogeneity was accomplished to a visually acceptable level. Adjustment was made for specific gravity of the vehicle. In order to obtain homogeneity, the test substance formulations were heated in a water bath with a maximum temperature of 50 °C for a maximum of 31 minutes. The test substance formulations were allowed to cool down below 40 °C prior to dosing.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: Not given in the report.
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
3 females per dose (same dose tested twice, so 6 females in total)
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Mortality/viability: Twice daily. Body weights: Days 1 (pre-administration), 8 and 15.
- Necropsy of survivors performed: yes
- Other examinations performed: Clinical signs: At periodic intervals on the day of dosing (day 1) and once daily thereafter, until day 15. The time of onset, degree and duration were recorded and the symptomgraded according to fixed scales.
Statistics:
N.A.
Sex:
female
Dose descriptor:
LD0
Effect level:
>= 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred.
Clinical signs:
Hunched posture was noted for all animals on day 1. In addition, lethargy and piloerection were noted among the majority of animals on day 1.
Body weight:
The body weight gain shown by the animals over the study period was considered to be similar to that expected of normal untreated animals of the same age and strain.
Gross pathology:
No abnormalities were found at macroscopic post mortem examination of the animals.
Other findings:
N.A.
Interpretation of results:
practically nontoxic
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Since no mortality occurred during the 14-day observation period, the LD50 of D-lactide exceeds 2000 mg/kg bw in female rats. Therefore, D-lactide is considered as not harmful upon ingestion.
Executive summary:

A sample of D-lactide (99 %) was examined for acute oral toxicity according to OECD guideline 423 with to two subsequent groups of three female Wistar rats (limit testing). A single dose level of 2000 mg/kg body weight was examined.

No mortality or distinct clinical signs, except hunched posture, piloerection and lethargy on day 1, were observed after treatment of 6 females with the 2000 mg/kg dose level. Macroscopic examination of the animals at the end of the observation period did not reveal any treatment-related gross changes. Since no mortality occurred during the 14-day observation period, the oral LD50 of D-lactide is considered to exceed 2000 mg/kg bw in female rats. D-lactide is considered as not harmful upon ingestion.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
1987-11-24 to 1988-01-08
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: GLP study according to OECD guideline 403. Lactic acid (termed SY-83 in the study) is used as read-across partner to DD-lactide.
Qualifier:
according to
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Version / remarks:
1981
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Fischer 344
Sex:
male/female
Details on test animals and environmental conditions:
Male and female Fischer 344 rats were obtained from Charles River Breeding Laboratories, Raleigh, North Carolina. Upon arrival, animals were quarantined for approximately 21 days. Stringent disease control procedures were followed during quarantine to assure the use of healthy animals. Rats were observed for signs of illness. The animals were judged to be healthy prior to utilization in this study and were 9-10 weeks old at initiation of exposure.
Animals were housed in an AAAIAC-accredited facility with a controlled environment. The temperature range was 71 ± 5 °F, while the humidity range was 50 ± 22 % with a brief excursion down to 18% during one day. The light cycle was maintained on a 12 hour light/dark cycle. Rats were individually housed in polycarbonate cages with filter tops and automatic watering devices. Corn-cob bedding was used and animals had free access to certified laboratory rodent chow which had been analyzed for environmental contaminants. Water and food were provided ad libitum, except during exposure.
Route of administration:
inhalation: aerosol
Type of inhalation exposure:
nose only
Vehicle:
other: unchanged (no vehicle)
Details on inhalation exposure:
Atmosphere generation:
A collison nebulizer (BGI Industries) was used to generate the aerosol. Compressed air was attached to the generator and the output from the aerosol was directed into a 4 liter dilution jar. Room air was allowed to dilute the aerosol before it was drawn into the exposure module under vacuum. A continuous, dynamic exposure to aerosolized test material was achieved with the nose-only exposure units. The air flow in the exposure module was adjusted to approximately 5 L/min using a calibrated rotameter and was periodically monitored during exposure. The exposure module consisted of a 360 ml cylindrical chamber mounted horizontally with an inlet, outlet and sampling port. Sampling of the test atmosphere was done from a sampling port situated on the side of the exposure module. Samples from the exposure module were taken for gravimetric and particle size determinations. Determinations of oxygen content could not be performed due to a malfunction of the oxygen sensor.
Conditions for animal exposures:
Rats were held in cylindrical tubes which aligned the heads of the animals with the conical shaped openings on the exposure chamber. The nose of each animal protruded into the chamber for nose-only inhalation. The animals were held in place by a soft sponge which served as a plunger in the cylinder and prevented the animal from turning or backing out. Five rats were restrained on each side of the exposure module. All rats of each concentration level were exposed at the same time.

Determination of aerosol concentration and particle size distribution:
Gravimetric determinations were made by pre-weighing a 24 mm diameter Whatman glass-fiber filter (GF/A), placing the filter in a filter holder, and connecting the filter to the sampling port of the exposure module. Air samples were drawn through the filter using a vacuum supply regulated by a flow meter. Flow rates were set 0.5 L/min and a sampling time of 4-8 min was used. The concentration of aerosol present in the chamber was calculated by the difference in weight of the filter divided by the volume of air sampled. Seven separate determinations were made and a time-weighted average was calculated for the total aerosol concentration.
Analytical verification of test atmosphere concentrations:
yes
Remarks:
Gravimetric, using a filter interception
Duration of exposure:
4 h
Concentrations:
7.94 mg/L
No. of animals per sex per dose:
5
Control animals:
yes
Details on study design:
Five male and five female rats were exposed to a nominal concentration of 7.94 mg/L of aerosolized test material for 4 hours. Analysis of at least three filter samples was performed to determine the purity of test material in the exposure module. The exposure atmosphere was also sampled to determine particle size distribution. A sham control group of 5 male and 5 female rats was used and was exposed to air alone for 4 hours. Animals were weighed prior to treatment and at weekly intervals thereafter. The animals were observed for mortality and pharmacotoxic signs during exposure, at 1 and 3 hours following exposure and once daily thereafter for 14 days. Survival was recorded for each group. Complete necropsies were performed on all animals on day 15 of the study. At necropsy, all organs showing gross lesions, if any, were fixed in 10% buffered formalin. Histopathology was not performed.
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 7.94 mg/L air
Exp. duration:
4 h
Mortality:
1 female died.
Clinical signs:
Animals were observed during exposure for signs of toxicity. Rapid breathing and eye tearing was observed in the treated group, while in the sham control group, respiration was calm and steady during exposure. One and three hours after exposure, the treated and control groups had a hunched posture, red stained fur surrounding the eyes (tearing), ruffled fur, and appeared ungroomed with soiled fur (stained brown). Female rats exposed to SY-83 appeared lethargic at one (2/5) and three hours (5/5). The two female rats that were lethargic at one hour also had rapid, shallow breathing and appeared to be gasping at both one and three hours. By 24 hours, most animals appeared normal and no unusual behavior or appearance was observed for the remainder of the test period. However, of the treated female rats, 4/5 had ruffled, ungroomed fur at 24 hours, and 3/5 had ruffled, ungroomed fur 2, 3 and 4 days after treatment. One female from the treated group had hunched posture, rapid and shallow breathing, and slight tremors, but these signs were observed only on day 5 post-treatment. One female rat from the treated group died on day 8 post-treatment. This animal was hunched with labored breathing and gasping on day 7. All other animals survived to the end of the study.
Body weight:
Individual body weights for animals on test are given in Tables 4 and 5. At the beginning of the study, mean body weights for individual groups were within 20 % of the overall mean for each sex. All groups of male rats gained weight within the first week after exposure in comparison to pre-exposure weights (3 % for sham-exposed, 2 % for SY-83, respectively). Female rats in the sham group gained weight during the first week after exposure (less than 1 %). Female rats in the treated group lost weight during the first week after exposure (7 %). After 14 days, all surviving animals had gained weight in comparison to pre-exposure weights (14 % for males, 7 % for females). No significant differences were observed in body weight between treated and control groups.
Gross pathology:
All surviving animals were necropsied at the termination of the study. The animal that died during the study was necropsied immediately. No gross lesions were observed at necropsy.
Interpretation of results:
practically nontoxic
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Under the conditions of the test, the LC50 of SY-83 is greater than 7.94 mg/L
Executive summary:

SY-83 was tested for its acute inhalation toxicity. Male and female F344 rats were exposed to a concentration of approximately 7.94 mg/L for 4 hours. Rapid breathing and eye tearing were observed during exposure. At one and three hours after exposure, all animals (including the sham controls) had a hunched posture, ruffled and ungroomed fur, brown stained fur and red-stained fur surrounding the eyes (tearing). By 24 hours, female treated rats had ruffled and stained coats. All other animals appeared normal at 24 hours and for the remainder of the 14 day observation period. Several treated female rats continued to have ruffled fur up to 4 days after exposure. One female rat from the treated group died on day 9. All other animals survived until the end of the study. Based on these results, the LC50 of SY-83 is greater than 7.94 mg/L. Lactic acid (termed SY-83 in the study) is used as read-across partner to DD-lactide.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LC50
7 940 mg/m³

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:
2010-05-28 to 2010-09-27
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP study
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
other: JMAFF Guidelines (2000), including the most recent revisions
Deviations:
no
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany.
- Age at study initiation: approximately 11 weeks old.
- Weight at study initiation:Body weight variation did not exceed ± 20 % of the sex mean (males: 312 g, females: 217 g).
- Fasting period before study:not applicable.
- Housing:Individually housed in labeled Macrolon cages (MIII type; height 18 cm) containing sterilised sawdust as bedding material (Litalabo, S.P.P.S., Argenteuil, France) and paper as cage-enrichment (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom).
- Diet (e.g. ad libitum): Free access to pelleted rodent diet (SM R/M-Z from SSNIFF@Spezialdiäten GmbH, Soest, Germany).
- Water (e.g. ad libitum): Free access to tap water.
- Acclimation period: At least 5 days before start of treatment under laboratory conditions. During the acclimatization period the animals were group housed in Macrolon cages (MIV type).

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.0 ± 3.0 °C (actual range: 19.8-21.5 °C)
- Humidity (%): relative humidity of 40-70 % (actual range: 38-75 %)
- Air changes (per h): approximately 15 air changes per hour
- Photoperiod (hrs dark/hrs light): 12 hours artificial fluorescent light and 12 hours darkness per day

IN-LIFE DATES: From09 June 2010 to 23 June 2010
Type of coverage:
occlusive
Vehicle:
polyethylene glycol
Details on dermal exposure:
TEST SITE
- Area of exposure: Approximately 10 % of the total body surface, i.e. approximately 25 cm² for males and 18 cm² for females.
- % coverage: Approximately 10 % of the total body surface.
- Type of wrap if used:The test substance formulation was held in contact with the skin with a dressing, consisting of a surgical gauze patch (Surgy 1D, Laboratoires Stella s.a., Liege, Belgium), successively covered with aluminium foil and Coban elastic bandage (3M, St. Paul, Minnesota, U.S.A. (Coban & Micropore)).

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The skin was cleaned of residual test substance using tap water.
- Time after start of exposure: 24 hours.

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg (10 ml/kg) body weight.

VEHICLE
Polyethylene glycol 400 (Merck, Darmstadt, Germany) (specific gravity 1.125). The vehicle was dehydrated before the formulation was prepared. The formulation (w/w) was prepared within 4 hours prior to dosing. Homogeneity was accomplished to a visually acceptable level. Adjustment was made for specific gravity of the vehicle. In order to obtain homogeneity, the test substance formulation was heated in a water bath with a temperature of 49.3 °C for 31 minutes. The test substance formulation was allowed to cool down below 40 °C prior to dosing.
Duration of exposure:
24 hours
Doses:
Single dosage, on day 1: 2000 mg/kg (10 ml/kg) body weight.
No. of animals per sex per dose:
5
Control animals:
not required
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Mortality/viability: Twice daily. Body weights: Days 1 (pre-administration), 8 and 15.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs: At periodic intervals on the day of dosing (day 1) and once daily thereafter, until day 15. The time of onset, degree and duration were recorded and the symptomgraded according to fixed scales.
Statistics:
N.A.
Sex:
male/female
Dose descriptor:
LD0
Effect level:
>= 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred.
Clinical signs:
Chromodacryorrhoea (left eye) was noted for one male on day 2.
One male showed scabs on day 6 and scales on day 7 in the treated skin area.
Body weight:
The changes noted in body weight gain in males and females were within the range expected for rats used in this type of study and were therefore considered not indicative of toxicity.
Gross pathology:
No abnormalities were found at macroscopic post mortem examination of the animals.
Interpretation of results:
practically nontoxic
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The dermal LD50 value of D-lactide in Wistar rats was established to exceed 2000 mg/kg body weight.
Executive summary:

In an acute dermal toxicity study (limit test), a group of young adult Wistar rats (5 males and 5 females) was dermally exposed to D-lactide (purity 99 %) in polyethylene glycol 400 for 24 hours to approximately 10 % of body surface area at 2000 mg/kg bw. Animals were observed for 14 days. No mortality occurred. There were no treatment related clinical signs, necropsy findings or changes in body weight. The dermal LD50 value of D-lactide in Wistar rats was established to exceed 2000 mg/kg body weight.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed

Additional information

After oral application of rats with D-lactide in accordance to OECD 423 an acute oral LD50 of D-lactide of > 2000 mg/kg bw was determined. After dermal application in accordance to OECD guideline 402 the LD50 of D-lactide was determined to be > 2000 mg/kg bw.

Lactic acid is an acceptable read-across partner as lactide is rapidly converted by hydrolysis into lactic acid under aqueous conditions.

In an acute inhalation toxicity study in rats with lactic acid in accordance to OECD guideline 403 an LC50 of > 7.94 mg/L air was determined.

Justification for selection of acute toxicity – oral endpoint
GLP guideline study

Justification for selection of acute toxicity – inhalation endpoint
In a GLP guideline study according to OECD 403 treated rats showed signs of toxicity and one female rat from the treated group died on day 9.

Justification for selection of acute toxicity – dermal endpoint
GLP guideline study

Justification for classification or non-classification

Based on the available data D-lactide does not warrant classification for acute toxicity. LD50 values for all routes are above the limit values of the relevant OECD guidelines.