Blue MGi 1037-Na (notification presscake)

Endpoint summary

Description of key information

Oral:

An acute oral toxicity study according OECD TG 423 was performed in rats with the test item (< 0.1% NaF). The oral LD50 in rats was determined to be > 2000 mg/kg bw.

In a further study according to OECD TG 423 the test item containing approx. 4.4% of the impurity NaF was administered to female rats. Here, the oral LD50 was determined to be > 300 to < 2000 mg/kg bw.

Dermal:

In a study performed with the read across substance Blue MGi 1037 in rats the LD50 was >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:

From 2016-11-02 to 2016-12-12

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

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Specific details on test material used for the study:

Purity: ca 88.0 %

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: TOXI COOP ZRT., 1103 Budapest, Hungary
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 8 weeks
- Weight at study initiation: 192 - 194 g (first step), 203 - 206 g (second step)
- Fasting period before study: overnight (diet)
- Housing: Group caging (3 animals/cage)
- Diet: ssniff® SM R/M-Z+H complete diet ad libitum, ssniff Spezialdiäten GmbH, D-59494 Soest, Germany
- Water: tap water ad libitum
- Acclimation period: 6 days in first step and 7 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: 12 hrs dark / 12 hrs light

IN-LIFE DATES: From: November 09, 2016 To: November 25, 2016

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 200 mg/mL
- Batch Number: 1608-5511

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

DOSAGE PREPARATION: Formulations were prepared just before the administration and stirred continuously during the treatment. The correction factor of 1.14 was taken into consideration for the preparation of the test item formulation.

CLASS METHOD
- Rationale for the selection of the starting dose: 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, so treatment with 2000 mg/kg bw was repeated on further three female rats. No animal died in the second step and thus the test was finished as the stopping criteria of Annex 2d of OECD Guideline No. 423 were met.

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

6 females

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: General state, external appearance, behavior and clinical symptoms after dosing at least once during the first 30 minutes, then 1 h, 2 h, 3 h, 4 h after the treatment and once each day (twice each day for mortality) for 14 days thereafter; body weights on day 0 (just before the treatment), on day 7 and on day 15
- Necropsy of survivors performed: Yes
- Other examinations performed: Macroscopic examination

Statistics:

Not applicable

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

act. ingr.

Mortality:

No deaths occurred after administration of 2000 mg/kg bw.

Clinical signs:

other: In group 1 clinical signs of reaction comprised of blue coloured faeces (5 cases of 57 observations) and diarrhoea (3/57). Blue coloured faeces (score +3) was detected in all animals on the treatment day between 2 and 4 hours after the treatment. Diarrho

Gross pathology:

Severe hydrometra was found in two females of the group 1 and in one female 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.

Other findings:

None

Interpretation of results:

GHS criteria not met

Conclusions:

The oral LD50 in rats was determined to be > 2000 mg/kg bw.

Executive summary:

An acute toxic class method according to OECD TG 423 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. No animal died in the second step, too, so the test was finished, the stopping criteria of Annex 2d of OECD Guideline No. 423 (presented in Appendix VII) was first met. Animals were weighed, observed for lethality and toxic symptoms for 14 days after the treatment. Gross pathological examination was carried out 15th day after the treatment.

No lethality was noted at single oral dose of 2000 mg/kg bw.

In the first step, disturbance of the autonomic functions (diarrhoea) was observed in one animal related with test item effect on the treatment day between 2 and 4 hours after the treatment. However, blue coloured faeces was detected in all animals on the treatment day between 2 and 4 hours after the treatment. This symptom was connected with the physical property of the test item.

In the second step, disturbance of the autonomic functions (diarrhoea) was observed in one animal related with test item effect on the treatment day between 3 and 4 hours after the treatment. However, blue coloured faeces was detected in all animals between the treatment day and Day 1. This symptom was connected with the physical property of the test item.

The body weight development was undisturbed in all animals.

All organs of the animals treated with 2000 mg/kg bw proved to be free of treatment related gross pathological changes.

The oral LD50 in rats was determined to be > 2000 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

OECD TG 423, GLP

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

the study does not need to be conducted because exposure of humans via inhalation is not likely taking into account the vapour pressure of the substance and/or the possibility of exposure to aerosols, particles or droplets of an inhalable size

Justification for type of information:

JUSTIFICATION FOR DATA WAIVING
The substance is of low volatility (vapour pressure 3.8 x 10-17 Pa).

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

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:

other: EEC Directive 92/69, B.3

Version / remarks:

1992

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: RCC Ltd, Biotechnology & Animal Breeding Division, Switzerland
- Age at study initiation: 10 weeks (males), 12 weeks (females)
- Weight at study initiation: males: 254.3-266.9 g / females: 191.7 -211.2g
- Housing: individually in Makrolon type-3 cages
- Diet: ad libitum, pelleted standard Kliba 3433
- Water: ad libitum, tap water
- Acclimation period: one week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22-24
- Humidity (%): 35-57
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

semiocclusive

Vehicle:

water

Remarks:

Bidistilled water

Details on dermal exposure:

TEST SITE
- % coverage: 10
- Type of wrap: semi-occlusive dressing fixed with an elastic adhesive bandage

REMOVAL OF TEST SUBSTANCE
- Washing: warm water
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 4 mL/kg bw

Duration of exposure:

24 h

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

yes, concurrent no treatment

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: test day 1,8, and 15 (weighting) and 1,2,3,5 hours, day 1 and twice daily on days 2-15 (mortality/viability)
- Necropsy of survivors performed: yes
- Clinical signs including body weight : yes

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No deaths occured during the study.

Clinical signs:

other: No clinical signs of toxicity were observed during the study period.

Gross pathology:

Effects on organs:
No macroscopic findings were observed at necropsy.

Interpretation of results:

GHS criteria not met

Conclusions:

In a study performed with Blue MGi 1037 in rats the LD50 was >2000 mg/kg bw.

Executive summary:

A group of five male and five female rats was treated with the substance Blue MGi 1037 at 2000 mg/kg by dermal application. The test item was suspended in bi-distilled water at a concentration of 0.5 g/mL and administered at a volume of 4 mL/kg. The animals were examined for clinical signs four times during test day 1 and once daily during test days 2-15. Mortality/viability was recorded together with clinical signs at the same time intervals on test day 1 and then twice daily on test days 2-15. Body weights were recorded on day 1 prior to administration and on days 8 and 15. All animals were necropsied and examined macroscopically.

No deaths occurred during the study. No clinical signs were observed during the observation period. Slight focal erythema were observed in 3 male and 3 female animals on test day 2 after removal of the dressing and persisted in 2 male animals until test day 5 and in 2 female animals until test days 3 and 4, respectively. Two female showed a marginal loss of body weight (1.7 and 1.8 %) one week after treatment. The body weight of the other animals was within the range commonly recorded for animals of this strain and age. No macroscopic findings were observed at necropsy.

The median lethal dose of the substance after single dermal administration to rats of both sexes, observed over a period of 14 days (LD50) is greater than 2000 mg/kg body weight.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

EEC Directive 92/69, B.3, GLP, Read Across

Additional information

Oral:

An acute toxic class method according to OECD TG 423 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. No animal died in the second step, too, so the test was finished, the stopping criteria of Annex 2d of OECD Guideline No. 423 (presented in Appendix VII) was first met. Animals were weighed, observed for lethality and toxic symptoms for 14 days after the treatment. Gross pathological examination was carried out 15th day after the treatment.

No lethality was noted at single oral dose of 2000 mg/kg bw.

In the first step, disturbance of the autonomic functions (diarrhoea) was observed in one animal related with test item effect on the treatment day between 2 and 4 hours after the treatment. However, blue coloured faeces was detected in all animals on the treatment day between 2 and 4 hours after the treatment. This symptom was connected with the physical property of the test item.

In the second step, disturbance of the autonomic functions (diarrhoea) was observed in one animal related with test item effect on the treatment day between 3 and 4 hours after the treatment. However, blue coloured faeces was detected in all animals between the treatment day and Day 1. This symptom was connected with the physical property of the test item.

The body weight development was undisturbed in all animals.

All organs of the animals treated with 2000 mg/kg bw proved to be free of treatment related gross pathological changes.

The LD50 of the test substance (< 0.1% NaF) was determined to be > 2000 mg/kg bw.

A further study according OECD TG 423 is available which was performed with the test substance that contained approx. 4.4% of the impurity NaF. Three groups, each of three female HanRcc:WIST (SPF) rats, were treated with Blue MGi 1037-Na (containing 4.4% NaF as impurity) by oral gavage administration one group at a dosage of 2000 mg/kg and two groups at 300 mg/kg body weight. The test item was diluted in vehicle (purified water) at a concentration of 0.2 or 0.03 g/mL, respectively and administered at a dosing volume of 10 mL/kg. The animals were examined daily during the acclimatization period and mortality, viability and clinical signs were recorded. All animals were examined for clinical signs at approximately 30 minutes, 1, 2, 3 and 5 hours after treatment on day 1 and once daily during test days 2-15. Mortality/viability was recorded at approximately 30 minutes, 1, 2, 3 and 5 hours after administration on test day 1 (with the clinical signs) and twice daily during days 2-15. Body weights were recorded on day 1 (prior to administration) and on days 8 and 15. All animals were necropsied and examined macroscopically. The following animals were treated and percentage of mortality was observed:

2 females treated at 2000 mg/kg       67 %

6 females treated at 300 mg/kg         0 %

Two animals died spontaneously at test day 2 in the 2000 mg/kg treatment group. All the other animals survived until the end of the study. The animals treated with 2000 mg/kg showed all a slight to moderate ruffled fur, a hunched posture and a slight to moderate sedation. These symptoms appeared all at the 30-minute reading and persisted in one animal until his death, in the other animal until the 3-hour reading or his death and in the surviving animal till the 3 -hour, 5 -hour or 3 -day reading. Dark blue watery faeces and a deep respiration were noted additionally in one animal between the 3-hour reading and his death. The surviving animal showed blue discolored faeces at test day 2 and 3.

All the animals treated with 300 mg/kg showed a slight ruffled fur from the 30-minute to the 3-hour reading. Blue faeces were present in all animals at test day 2 and persisted in 3 animals until the 3-day reading. In addition a hunched posture was noted in 3 animals between 30 minutes and 3 hours after treatment.

One animal showed a loss of body weight (approximately 9%) at the end of the observation period. The body weight of all the other animals was within the range commonly recorded for this strain and age.

The two animals which died spontaneously showed a blue discoloration of the stomach, duodenum, jejunum, ileum, caecum and colon.

The median lethal dose of Blue MGi 1037-Na (with appr. 4.4% NaF as impurity) after single oral administration to female rats is: 300 mg/kg body weight <LD50 (female rat): < 2000 mg/kg body weight.

Dermal:

A group of five male and five female rats was treated with the read across substance Blue MGi 1037 at 2000 mg/kg by dermal application. The test item was suspended in bi-distilled water at a concentration of 0.5 g/mL and administered at a volume of 4 mL/kg. The animals were examined for clinical signs four times during test day 1 and once daily during test days 2-15. Mortality/viability was recorded together with clinical signs at the same time intervals on test day 1 and then twice daily on test days 2-15. Body weights were recorded on day 1 prior to administration and on days 8 and 15. All animals were necropsied and examined macroscopically.

No deaths occurred during the study. No clinical signs were observed during the observation period. Slight focal erythema were observed in 3 male and 3 female animals on test day 2 after removal of the dressing and persisted in 2 male animals until test day 5 and in 2 female animals until test days 3 and 4, respectively. Two female showed a marginal loss of body weight (1.7 and 1.8 %) one week after treatment. The body weight of the other animals was within the range commonly recorded for animals of this strain and age. No macroscopic findings were observed at necropsy.

The median lethal dose of the substance after single dermal administration to rats of both sexes, observed over a period of 14 days (LD50) is greater than 2000 mg/kg body weight.

Justification for classification or non-classification

Classification, Labeling, and Packaging Regulation (EC) No 1272/2008

The available data for acute toxicity are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. Based on this data, the substance containing <0.1% NaF is not subject to classification for acute oral and dermal toxicity under Regulation (EC) No 1272/2008, as amended for the twelfth time in Regulation (EU) 2019/521. However, the substance which contains approx. 4.4% of the impurity NaF is classified for acute oral toxicity Cat.4 (H302).