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Administrative data

Description of key information

The test material did not cause any adverse effects after singel oral or dermal administration to rats at a dose level of 2000 mg/kg bw.

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:
2012-07-16 to 2012-11-15
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline 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
GLP compliance:
yes (incl. certificate)
Remarks:
(Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, München, Germany)
Test type:
acute toxic class method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
Housing and Feeding Conditions
- Full barrier in an air-conditioned room
- Temperature: 22 +/- 3 °C
- Relative humidity: 55 +/- 10%
- Artificial light, sequence being 12 hours light, 12 hours dark
- Air change: 10 x / hour
- Free access to Altromin 1324 maintenance diet for rats and mice
- Free access to tap water, sulphur acidified to a pH value of approximately 2.8 (drinking water, municipal residue control, microbiological controls at regular intervals)
- The animals were kept in groups / individually in IVC cages, type III H, polysulphone cages on Altromin saw fibre bedding
- Certificates of food, water and bedding are filed at BSL BIOSERVICE
- Adequate acclimatisation period (at least five days) under laboratory conditions
Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
The animals were marked for individual identification by tail painting.
Prior to the administration a detailed clinical observation was made of all animals.
Prior to the administration food was withheld from the test animals for 16 to 19 hours (access to water was permitted). Following the period of fasting the animals were weighed and the test item was administered. Food was provided again approximately 4 hours post dosing.
The test item was administered at a single dose by gavage using a feeding tube.
The test item was administered at a dose volume of 10 mL/kg body weight.
Doses:
Dose Level:
The starting dose was selected to be 2000 mg/kg body weight. No compound-related mortality was recorded for any animal of step 1 or 2. Based on these results and according to the acute toxic class method regime no further testing was required.
No. of animals per sex per dose:
6
Control animals:
no
Details on study design:

All animals were observed for 14 days after dosing for general clinical signs, morbidity and mortality.
The animals were weighed on day 1 (prior to the administration) and on days 8 and 15. A careful clinical examination was made several times on the day of dosing (at least once during the first 30 minutes and with special attention given during the first 4 hours post-dose). As soon as symptoms were noticed they were recorded. Thereafter, the animals were observed for clinical signs once daily until the end of the observation period. All abnormalities were recorded. Cageside observations included changes in the skin and fur, eyes and mucous membranes. Also respiratory, circulatory, autonomic and central nervous systems and somatomotor activity and behaviour pattern were examined. Particular attention was directed to observations of tremor, convulsions, salivation, diarrhoea, lethargy, sleep and coma. At the end of the observation period the surviving animals were sacrificed with an overdosage of pentobarbital injected intraperitoneally (Narcoren®, Merial; lot no.: 221022; expiry date: 02/2015) at a dosage of approximately 8 mL/kg bw. All animals were subjected to gross necropsy. All gross pathological changes were recorded.
Statistics:
According to OECD guidelines, the biological relevance of the results is the criterion for the interpretation of results, a statistical evaluation of the results is not regarded as necessary.
Sex:
female
Dose descriptor:
approximate LD50
Effect level:
> 2 000 mg/kg bw
Mortality:
All animals survived until the end of the study without showing any signs of toxicity.
Clinical signs:
None.
Body weight:
Throughout the 14-day observation period, the body weight gain of the test animals was within the normal range of variation for this strain.
Gross pathology:
At necropsy, no macroscopic findings were observed in any animal of any step.

Table: Clinical Signs - Individual Data

Animal
No. / Sex

Time of
Observation
Post-Dose

Observations

Step 1 (2000 mg/kg Body Weight)

1 / female

during the whole observation period

no signs of toxicity

2 / female

during the whole observation period

no signs of toxicity

3 / female

during the whole observation period

no signs of toxicity

Step 2 (2000 mg/kg Body Weight)

4 / female

during the whole observation period

no signs of toxicity

5 / female

during the whole observation period

no signs of toxicity

6 / female

during the whole observation period

no signs of toxicity

Table: Body Weight Development - Absolute Body Weights in g and Body Weight Gain in %

Animal No. /
Sex

g
Day 1

g
Day 8

g
Day 15

%
Day 1-15

Step 1 (2000 mg/kg Body Weight)

1 / female

166

191

197

19

2 / female

183

204

226

23

3 / female

170

195

206

21

Step 2 (2000 mg/kg Body Weight)

4 / female

171

192

200

17

5 / female

159

177

185

16

6 / female

162

185

197

22

Table: Findings of Necropsy - Individual Data

Animal No./
Sex

Organ

Macroscopic Findings

Step 1 (2000 mg/kg Body Weight)

1 / female

-

nsf

2 / female

-

nsf

3 / female

-

nsf

Step 2 (2000 mg/kg Body Weight)

4 / female

-

nsf

5 / female

-

nsf

6 /female

-

nsf

nsf = no specific findings

Table: LD50 Cut-Off

Dose
(unit)

Number of
Animals
Investigated

Number of Intercurrent Deaths

LD50 Cut-Off

2000 mg/kg bw

6

0

unclassified

bw = body weight

Conclusions:
Under the conditions of the present study, a single oral application of the test item Fatty acids, C16-18-, reaction products with diethanolamine to rats at a dose of 2000 mg/kg body weight was associated with no signs of toxicity or mortality.
The median lethal dose of Fatty acids, C16-18-, reaction products with diethanolamine after a single oral administration to female rats, observed over a period of 14 days is:
LD50 cut-off (rat): unclassified
Executive summary:

1.1.           Summary Results

Two groups, each of three female WISTAR Crl: WI(Han) rats, were treated with the test item by oral gavage administration at a dosage of 2000 mg/kg body weight. The test item was suspended in the vehicle aqua ad injectionem (sterile water) at a concentration of 0.2 g/mL and administered at a dose volume of 10 mL/kg.

All animals used in the study after their entrance at BSL were allowed to acclimatise to the laboratory conditions for at least 5 days. The animals were observed on delivery, on inclusion in the study and before administration for mortality/morbidity and other clinical signs. All animals were examined for clinical signs several times on the day of dosing and once daily until the end of the observation period. Their body weights were recorded on day 1 (prior to the administration) and on days 8 and 15.All animals were necropsied and examined macroscopically.

Table: Results per Step

Step

Sex/No.

Dose (mg/kg)

Number of Animals

Number of Intercurrent Deaths

1

female/1-3

2000

3

0

2

female/4-6

2000

3

0

All animals survived until the end of the study without showing any signs of toxicity.

Throughout the 14-day observation period, the body weight gain of the test animals was within the normal range of variation for this strain.

At necropsy, no macroscopic findings were observed in any animal of any step.

LD50cut-off:                                                     unclassified

Species/strain:                                                   WISTAR Crl: WI(Han) rats

Number of animals:                                           3 per step / 2 steps performed

Vehicle:                                                            aqua ad injectionem

Method:                                                             OECD 423
          EC 440/2008, Method B.1 tris
          OPPTS 870.1000
          OPPTS 870.1100


 

Conclusion

Under the conditions of the present study, a single oral application of the test item Fatty acids, C16-18-, reaction products with diethanolamine to rats at a dose of 2000 mg/kg body weight was associated with no signs of toxicity or mortality.

The median lethal dose of Fatty acids, C16-18-, reaction products with diethanolamine after a single oral administration to female rats, observed over a period of 14 days is:

LD50cut-off (rat): unclassified

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
2 000 mg/kg bw
Quality of whole database:
Klimisch 1; recent study according to current guideline

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:
2012-08-23 to 2012-11-16
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline 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
GLP compliance:
yes (incl. certificate)
Remarks:
(Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, München, Germany)
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
- Full barrier in an air-conditioned room
- Temperature: 22 +/- 3 °C
- Relative humidity: 55 +/- 10%
- Artificial light, sequence being 12 hours light, 12 hours dark
- Air change: 10 x / hour
- Free access to Altromin 1324 maintenance diet for rats and mice (lot no. XX)
- Free access to tap water, sulphur acidified to a pH value of approximately 2.8 (drinking water, municipal residue control, microbiological controls at regular intervals)
- The animals were kept individually in IVC cages, type III H, polysulphone cages on Altromin saw fibre bedding (lot no. XX)
- Certificates of food, water and bedding are filed at BSL BIOSERVICE
- Adequate acclimatisation period (at least five days) under laboratory conditions
Vehicle:
water
Details on dermal exposure:
Preparation of the Animals:

The animals were marked for individual identification by tail painting.
Approximately 24 hours before the test, the fur was removed from the dorsal area of the trunk using an electric clipper. Care was taken to avoid abrading the skin, and only animals with healthy intact skin were used.
No less than 10% of the body surface was cleared for the application.
Prior to the application a detailed clinical observation was made of all animals.

Application:

The test item was applied at a single dose, uniformly over an area which was approximately 10% of the total body surface.
The test item was held in contact with the skin by a dressing throughout a 24-hour period. The dressing consisted of a gauze-dressing and non-irritating tape and was fixed with an additional dressing in a suitable manner.
Duration of exposure:
The test item was held in contact with the skin throughout a 24-hour period. At the end of the exposure period the residual test item was removed using aqua ad injectionem.
Doses:
The test item was applied at a single dose of 2000 mg/kg body weight to each animal.
No. of animals per sex per dose:
5 male and 5 female
Control animals:
not required
Details on study design:
Observation period:

All animals were observed for 14 days after dosing

Weight Assessment:

The animals were weighed on day 1 (prior to the application) and on days 8 and 15.

Clinical Examination:

careful clinical examination was made several times on the day of dosing (at least once during the first 30 minutes and with special attention given during the first 4 hours post-dose). As soon as symptoms were noticed they were recorded. Thereafter, the animals were observed for clinical signs once daily until the end of the observation period. All abnormalities were recorded.
Cageside observations included changes in the skin and fur, eyes and mucous membranes. Also respiratory, circulatory, autonomic and central nervous systems and somatomotor activity and behaviour pattern were examined. Attention was directed to observations of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.

Pathology:

At the end of the observation period the surviving animals were sacrificed with an overdosage of pentobarbital injected intraperitoneally (Narcoren®, Merial) at the dosage of approximately 8 mL/kg bw. All animals were subjected to gross necropsy. All gross pathological changes were recorded.

Evaluation of Results:

Individual reactions of each animal were recorded at each time of observation.
Toxic response data were recorded by sex and dose level.
Nature, severity and duration of clinical observations were described.
The body weight changes were summarised in a tabular form.
Necropsy findings were described.


Statistics:
According to OECD guidelines, the biological relevance of the results is the criterion for the interpretation of results, a statistical evaluation of the results is not regarded as necessary.
Dose descriptor:
approximate LD50
Effect level:
2 000 mg/kg bw
Based on:
test mat.
Mortality:
None
Clinical signs:
No treatment-related effects were observed.
Body weight:
The body weight development of all male and female animals was within the expected range.
Gross pathology:
No treatment-related effects were observed.
Other findings:
No erythema or oedema was observed. Scratches were observed in 1 of 5 male and female animals.
All findings were reversible within the observation period.
Interpretation of results:
practically nontoxic
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Under the conditions of the present study, single dermal application of the test item Fatty acids, C16-18-, reaction products with diethanolamine to rats at a dose of 2000 mg/kg body weight was associated with neither mortality nor signs of toxicity but slight signs of irritation.
The dermal LD50 was determined to be > 2000 mg Fatty acids, C16-18-, reaction products with diethanolamine / kg body weight.
Executive summary:

Summary Results:

LD50:                                                        > 2000 mg /kg bw

Species/strain:                                         WISTAR Crl: WI(Han) rats

Vehicle (moistening):                               aqua ad injectionem

Number of animals:                                  5 male and 5 female

Duration of exposure:                             24 hours

Method:                                                  OECD 402[3]
EC 440/2008, Method B.3[4]
OPPTS 870.1200[5]

Table:  Results per Step

Sex

Dose
(mg/kg bw)

Number
of Animals

Number
of Intercurrent Deaths

male

2000

5

0

female

2000

5

0

Signs of toxicity related to dose level used, time of onset and duration:

No treatment-related effects were observed.

Effect on organs (related to dose level):

No treatment-related effects were observed.

Signs of irritation:

No erythema or oedema was observed. Scratches were observed in 1 of 5 male and female animals.

All signs of irritation were reversible within the observation period.

Conclusion:

Under the conditions of the present study, single dermal application of the test item Fatty acids, C16-18-, reaction products with diethanolamine to rats at a dose of 2000 mg/kg body weight was associated with neither mortality nor signs of toxicity but slight signs of irritation.

The dermal LD50 was determined to be > 2000 mg Fatty acids, C16-18-, reaction products with diethanolamine / kg body weight.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
2 000 mg/kg bw
Quality of whole database:
Klimisch 1; recent study according to current guideline

Additional information

Acute exposure of rats via oral or dermal route did not result in any adverse systemic or local effects.

Justification for classification or non-classification

Not classified

Neither after oral nor after dermal exposure any mortalities were detected at 2000 mg/kg bw. Therefore criteria for classification are not fullfilled.