Reaction mass of lauric acid, compound with morpholine (1:1) and 2-ethylhexyl dihydrogen phosphate, compound with morpholine (1:2) and bis(2-ethylhexyl) hydrogen phosphate, compound with morpholine (1:1)

Endpoint summary

Description of key information

Based on the data available the test item is considered to not be classified as acute toxic by dermal and oral route.

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:

2016-03-01 to 2016-04-19

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)

Version / remarks:

2001

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Version / remarks:

2008

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Version / remarks:

2002

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Toxi-Coop Zrt. 1103 Budapest, Cserkesz u. 90
- Females nulliparous and non-pregnant: yes
- Age at study initiation: Young adult rats, 8 weeks old in group 1 and group 2
- Weight at study initiation: 147-149 g (group 1), 147-156 g (group 2)
- Fasting period before study:
- Housing: 3 animals/sex/cage, cage type: Type II polypropylene/polycarbonate; rat type cages with a solid floor, stainless steel wire covers and self-feeding baskets.
- Diet: The animals received ssniff® SM R/M-Z+H complete diet produced by ssniff Spezialdiäten GmbH, D-59494 Soest Germany, ad libitum. The food is periodically analysed and is considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study. Copies of the relevant Certificates of Analysis are maintained in Toxi-Coop Zrt.’s archive.
- Water: The drinking water is periodically analysed and is considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study. Copies of the relevant Certificates of Analysis are maintained in Toxi-Coop Zrt.’s archive.
- Acclimation period: 5 days in first step and 6 days in second step.

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 3 °C
- Humidity: 30 - 70 %
- Air changes: above 10 air exchanges/hour by central air-condition system.
- Photoperiod: Artificial light, from 6 am. to 6 pm.

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 200 mg/mL
- Lot/batch no.: 1511-5526
- Purity: Aqua purificata
- Expiry date: 26 May 2016
- Manufactured: Parma Produkt Kft.

MAXIMUM DOSE VOLUME APPLIED: The dose was formulated in the vehicle. Concentration of formulations were adjusted to maintain a treatment volume of 10 mL/kg bw.

CLASS METHOD
- Rationale for the selection of the starting dose: The starting dose was selected on the basis of the available information about the test item.

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

2 x 3

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Inspection for signs of morbidity and mortality were made twice daily at the beginning and end of the working day. Animals were observed individually after dosing once during the first 30 minutes, then 1 h, 2 h, 3 h, 4 h, after the treatment and once per day for 14 days thereafter. Individual observations were performed on the skin and fur, eyes and mucous membranes and also respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma. The body weight were recorded on day 0 (shortly before the treatment), on day 7 and on day 15 on all animals with a precision of 1 g, respectively.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, gross pathology

Key result

Sex:

female

Dose descriptor:

discriminating dose

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

The test item Madurit TM 3750 did not induce mortality following a single oral administration to female rats at a dose of 2000 mg/kg bw. All female rats survived the performed treatment until the end of the 14-day observation period.

Clinical signs:

other: In group 1 treated with 2000 mg/kg bw no systemic toxic symptoms were observed throughout the 14-day post-treatment period. In group 2 treated with 2000 mg/kg bw clinical signs of toxicity comprised of decreased activity (8 cases out of 57 observations),

Gross pathology:

All animals treated with 2000 mg/kg bw dose of test item survived until the scheduled necropsy on Day 15. Severe hydrometra was found in female No.: 1086 of the group 1 and in two females (No.: 1098, 1097) of group 2. Hydrometra is physiological finding and connected to the cycle of the animal. No pathological changes were found related to the effect of the test item during the macroscopic examination of animals.

Interpretation of results:

GHS criteria not met

Conclusions:

The method used is not intended to allow for the calculation of a precise LD50 value. However for this acute oral toxicity study with the test item in rats the determined LD50 is above 2000 mg/kg bw.

Executive summary:

An acute oral toxicity study was carried out using the class method according to OECD guideline 423. The starting dose was selected on the basis of the available information about the test item. The acute toxic class method was carried out involving a stepwise procedure with the use of 2000 mg/kg bw as the starting dose in three female rats. No animal died in the first step at 2000 mg/kg bw dose level. Therefore, treatment with 2000 mg/kg bw was repeated on further three female rats. Again, no animal died in the second step, thus, no further testing was required. The stopping criteria of Annex 2d of OECD Guideline No. 423 were met. Animals were weighed, observed for lethality and toxic symptoms for 14 days after the treatment. Gross pathological examination was carried out on 15th day after the treatment. No lethality was noted following oral administration of a single dose of 2000 mg/kg bw. In the first step at a dose level of 2000 mg/kg bw no clinical symptoms were observed on the day of the treatment and during the 14-day observation period, the general state and behaviour of experimental animals were normal. In the second step at a dose level of 2000 mg/kg bw, CNS - and emotion symptom (decreased activity) and disturbances of the autonomic functions (salivation, piloerection) were observed in animals on the treatment day between 30 minutes and 3 hours after the treatment. The body weight development was normal in all animals. All organs of the animals treated with 2000 mg/kg bw dose proved to be free of treatment related gross pathological changes.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

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:

2016-03-02 to 2016-04-11

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Version / remarks:

1987

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2008

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Version / remarks:

1998

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Toxi-Coop Zrt. 1103 Budapest, Cserkesz u. 90
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: Young adult rats
- Weight at study initiation: 200-219 g
- Fasting period before study: not applicable
- Housing: During acclimatisation: 3 animals/sex/cage During the study: animals were housed individually. Cage type: Type II polypropylene/polycarbonate; rat type cages with a solid floor, stainless steel wire covers and self-feeding baskets.
- Diet: The animals received ssniff® SM R/M-Z+H complete diet produced by ssniff Spezialdiäten GmbH, D-59494 Soest Germany, ad libitum. The food was periodically analysed and was considered free from any contaminants that could have affected the purpose or integrity of the study. Copies of the relevant Certificates of Analysis are maintained in Toxi-Coop Zrt.’s archive.
- Water: Animals received tap water from watering bottles ad libitum. The drinking water was periodically analysed and was considered free from any contaminates that could have affected the purpose or integrity of the study. Copies of the relevant Certificates of Analysis are maintained in Toxi-Coop Zrt.’s archive.
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 3 °C
- Humidity: 30 - 70 %
- Air changes: Above 10 air exchanges/hour by central air-condition system.
- Photoperiod: Artificial light, from 6 am. to 6 pm.

Type of coverage:

semiocclusive

Vehicle:

water

Details on dermal exposure:

TEST SITE
- Area of exposure: approx. 10 % of total body surface area, at the back
- % coverage: at least 10% area of the total body surface
- Type of wrap if used: Sterile gauze pads were placed on the skin of rats. These gauzes were kept in contact with the skin by a patch with adhesive hypoallergenic plaster. The entire trunk of the animal was wrapped with semi-occlusive plastic wrap for 24 hours.

REMOVAL OF TEST SUBSTANCE
- Washing: At the end of the exposure period, residual test item was removed, using body temperature water.
- Time after start of exposure: 24 h

TEST MATERIAL
- Amount applied: 2000, 300, 50, 5 mg/kg bw
- Concentrations: 400, 60, 10, 1 mg/ml
- Constant volume or concentration used: yes. 5 mL/kg bw of respective concentration
- For solids, paste formed: yes

VEHICLE
- Lot/batch no.: 1511-5526
- Purity: Aqua purificata
- Date of expiration: 26 May 2016
- Supplier: Parma Produkt Kft.

Duration of exposure:

24 h

Doses:

2000, 300, 50, 5 mg/kg bw

No. of animals per sex per dose:

2

Control animals:

not required

Details on study design:

- Duration of observation period following administration: 7 days
- Frequency of observations and weighing: Inspection for signs of morbidity and mortality were made twice daily at the beginning and end of the working day. Animals were observed individually 1 h and 5 h after dosing, and once each day for 7 days thereafter. Observations included the skin and fur, eyes and mucous membranes, and also respiratory, circulatory, autonomic and central nervous system, and somatomotor activity and behaviour pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma. The body weights were recorded on day 0 (shortly before the treatment) and on Day 7 (with a precision of 1 g).
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, gross pathology

Key result

Sex:

female

Dose descriptor:

discriminating dose

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred after the 24-hour dermal exposure to Madurit TM 3750 in HsdHan:WIST female rats during the preliminary study.

Clinical signs:

other: No systemic toxic signs were noted during the study in all doses. In group 1 treated with 2000 mg/kg bw dermal corrosive symptoms as erythema and other signs as necrosis, desquamation and wound were observed in both animals on the treatment site. Severe r

Gross pathology:

All animals survived until the scheduled necropsy on Day 7. In group 1 (2000 mg/kg bw) external macroscopic changes as necrosis, desquamation and wound were observed on the treatment site in both animals (No.: 1109, 1125). In group 2 (300 mg/kg bw) external macroscopic changes as dry and desquamated skin were observed on the treatment site in both animals (No.: 1122, 1123). Internal necropsy finding as pale liver was found in animal No.: 1123. Severe hydrometra was recorded in animal No.: 1122. In group 3 (50 mg/kg bw) internal macroscopic change as pale liver was observed in animal No.: 1121. In group 4 (5 mg/kg bw) severe hydrometra was observed in animal No.: 1124. No macroscopic alterations due to the systemic toxic effects of the test item were found.

Interpretation of results:

GHS criteria not met

Conclusions:

In this preliminary acute dermal toxicity study with the test item Madurit TM 3750, the obtained acute dermal LD50 value was greater than 2000 mg/kg bw in female HsdHan:WIST rats. According to Regulation (EC) No 1272/2008 and UN GHS, the test item has not been classified into any category. On the other hand, it is to be noted that the test item caused dermal corrosion/irritation response on the site of administration.

Executive summary:

An acute dermal toxicity study was performed with the test item in HsdHan:WIST female rats, in compliance with OECD Guideline No. 402 and OPPTS 870.1200. Only a preliminary study was carried out because of severe skin corrosive effect of the test item observed in the 2000 mg/kg bw dose group. Altogether four groups (n=2 animals) was exposed to the test item at 2000, 300, 50 and 5 mg/kg bw by dermal route. The test item was applied in diluted form and left in contact with the skin for 24 hours, followed by a 7-day observation period. The vehicle was distilled water. The results of the preliminary study were summarised as follows: No mortality occurred after the 24-hour dermal exposure to the test item in all groups during the study. No systemic toxic signs were noted during the study in all doses. The test item caused local dermal corrosion/irritation symptoms on the treatment site. Severe erythema, necrosis, desquamation and wound occurred in the 2000 mg/kg bw dose group between Day 1 and Day 7. Very slight to well defined redness, dry skin and desquamation were observed in the300 mg/kg bw dose group between Day 1 and Day 7. Well defined redness was observed in the 50 mg/kg bw dose group between Day 1 and Day 2. No local dermal symptoms were found in the 5 mg/kg bw dose. Mean body weight development was within the normal range for all animals of this strain and age. It is to be noted that a backwardness of body weight gain was observable in the 2000 mg/kg bw dose compared to other groups. It is explicable with the marked distress and pain due to corrosive effect of test item. No macroscopic alterations of organs and tissues referred to the systemic toxic effect of the test item were seen during the necropsy. In this preliminary acute dermal toxicity study with the test item Madurit TM 3750, the obtained acute dermal LD50 value was greater than 2000 mg/kg bw in female HsdHan:WIST rats. According to Regulation (EC) No 1272/2008 and UN GHS, the test item has not been classified into any category. On the other hand, it is to be noted that the test item caused dermal corrosion/irritation response on the site of administration.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

GLP and guideline study

Additional information

Oral route:

An acute oral toxicity study was carried out using the class method according to OECD guideline 423. The starting dose was selected on the basis of the available information about the test item. The acute toxic class method was carried out involving a stepwise procedure with the use of 2000 mg/kg bw as the starting dose in three female rats. No animal died in the first step at 2000 mg/kg bw dose level. Therefore, treatment with 2000 mg/kg bw was repeated on further three female rats. Again, no animal died in the second step, thus, no further testing was required. The stopping criteria of Annex 2d of OECD Guideline No. 423 were met. Animals were weighed, observed for lethality and toxic symptoms for 14 days after the treatment. Gross pathological examination was carried out on 15th day after the treatment. No lethality was noted following oral administration of a single dose of 2000 mg/kg bw. In the first step at a dose level of 2000 mg/kg bw no clinical symptoms were observed on the day of the treatment and during the 14-day observation period, the general state and behaviour of experimental animals were normal. In the second step at a dose level of 2000 mg/kg bw, CNS - and emotion symptom (decreased activity) and disturbances of the autonomic functions (salivation, piloerection) were observed in animals on the treatment day between 30 minutes and 3 hours after the treatment. The body weight development was normal in all animals. All organs of the animals treated with 2000 mg/kg bw dose proved to be free of treatment related gross pathological changes.

Dermal route:

An acute dermal toxicity study was performed with the test item in HsdHan:WIST female rats, in compliance with OECD Guideline No. 402 and OPPTS 870.1200. Only a preliminary study was carried out because of severe skin corrosive effect of the test item observed in the 2000 mg/kg bw dose group. Altogether four groups (n=2 animals) was exposed to the test item at 2000, 300, 50 and 5 mg/kg bw by dermal route. The test item was applied in diluted form and left in contact with the skin for 24 hours, followed by a 7-day observation period. The vehicle was distilled water. The results of the preliminary study were summarised as follows: No mortality occurred after the 24-hour dermal exposure to the test item in all groups during the study. No systemic toxic signs were noted during the study in all doses. The test item caused local dermal corrosion/irritation symptoms on the treatment site. Severe erythema, necrosis, desquamation and wound occurred in the 2000 mg/kg bw dose group between Day 1 and Day 7. Very slight to well defined redness, dry skin and desquamation were observed in the300 mg/kg bw dose group between Day 1 and Day 7. Well defined redness was observed in the 50 mg/kg bw dose group between Day 1 and Day 2. No local dermal symptoms were found in the 5 mg/kg bw dose. Mean body weight development was within the normal range for all animals of this strain and age. It is to be noted that a backwardness of body weight gain was observable in the 2000 mg/kg bw dose compared to other groups. It is explicable with the marked distress and pain due to corrosive effect of test item. No macroscopic alterations of organs and tissues referred to the systemic toxic effect of the test item were seen during the necropsy. In this preliminary acute dermal toxicity study with the test item, the obtained acute dermal LD50 value was greater than 2000 mg/kg bw in female HsdHan:WIST rats. According to Regulation (EC) No 1272/2008 and UN GHS, the test item has not been classified into any category. On the other hand, it is to be noted that the test item caused dermal corrosion/irritation response on the site of administration.

Justification for classification or non-classification

Classification, Labelling, and Packaging Regulation (EC) No 1272/2008
The available experimental test data are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. Based on available data on acute toxicity, the test item is not classified according to Regulation (EC) No 1272/2008 (CLP), as amended for the eighth time in Regulation (EU) No 2016/918.