4H-Pyrazino[2,1-a]isoquinolin-4-one, 2-acetyl-1,2,3,6,7,11b-hexahydro-

Endpoint summary

Description of key information

Acute oral toxicity in the rat according to OECD TG 423 revealed that 300 < LD50 < 2000 mg/kg bw (reference 7.2.1 -1).

Acute dermal toxicity in the rat according to OECD TG 402 revealed an LD50 > 2000 mg/kg bw (reference 7.2.3 -1).

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:

2004-11-09 to 2005-02-11

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: F. Winkelmann, 33178 Borchen
- Females nulliparous and non-pregnant: not specified
- Age at study initiation: young adult
- Weight at study initiation: 151 - 172 g
- Fasting period before study: yes
- Housing: individually
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22- 23 °C
- Humidity: 43 - 59 %
- Photoperiod: 12 / 12 hrs dark / hrs light

Route of administration:

oral: gavage

Vehicle:

methylcellulose

Remarks:

aqueous Methocel K4M Premium solution

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 30 and 200 g/L
- Amount of vehicle: 10 mL/kg
- Justification for choice of vehicle: meets the formulation standards set by the USP, JP and Ph. Eur.

MAXIMUM DOSE VOLUME APPLIED: 2000 mg/kg bw

Doses:

300, 2000 mg/kg body weight

No. of animals per sex per dose:

3 per sex per dose @ 300 mg/kg bw
3 females @ 2000 mg/kg bw

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: on days 2, 4, 6, 8, 11, 13, and 15
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, gross pathological examination

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 300 - < 2 000 mg/kg bw

Based on:

test mat.

Mortality:

Male: 300 mg/kg bw; Number of animals: 3; Number of deaths: 0
Female: 300 mg/kg bw; Number of animals: 3; Number of deaths: 0
Female: 2000 mg/kg bw; Number of animals: 3; Number of deaths: 3

Clinical signs:

other: Signs of toxicity related to dose levels: After oral treatment with 300 mg/kg (3 males, 3 females), signs of toxicity were seen in the male and female. They started 1 to 15 minutes after administration and lasted up to 24 hours. They consisted of locomo

Gross pathology:

Effects on organs: For animals treated with 300 mg/kg body weight, no organ alterations were seen at necropsy The female rat (2000 mg/kg bw), which died, showed changes in the gastrointestinal tract. The two other rats, which were sacrified in a moribund state showed no macroscopic abnormalities.

Table 1 Summary of mortalities

Dose (mg/kg bw)	Dead / treated animals
	males	Females
300	0/3	0/3
2000	-	3/3

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

According to the results of this study, the LD50 value of the test material is expected to be between 300 - 2000 mg/kg bw.

Executive summary:

Study design

The acute toxicity of the test material was determined in rats after oral administration of 300 and 2000 mg/kg body weight. Directly before administration the test material was prepared with aqueous Methocel® K4M Premium solution as the vehicle. This study was performed according to the "Acute toxic class method" (ATC) as described in the OECD Guideline 423.

Results

After oral treatment with 300 mg/kg bw (3 males, 3 females), signs of toxicity were seen in the male and female rats. They started 1 to 2 hours after dosing and lasted up to 24 hours. They consisted of locomotor disturbance and dyspnea. All these animals survived the observation period. All rats, which were sacrificed at the end of the study, showed no macroscopic abnormalities.
After oral treatment with 2000 mg/kg bw (3 females), signs of toxicity were seen in the female rats. They started 1 to 15 minutes after administration and lasted up to 24 hours. They consisted of incomplete eyelid closure, abdominal position, respiratory sounds, locomotor disturbance, and dyspnea. One animal died 6 hours after administration. The two other animals were sacrificed in a moribund state 8 hours after administration.
The female rat (2000 mg/kg bw), which died, showed changes in the gastrointestinal tract (liquid contents of the stomach and thickened contents of the cecum). The other two rats, which were sacrificed in moribund state showed no macroscopic abnormalities.

Conclusions

According to the results of this study, the LD50 value of the test material is expected to be between 300 - 2000 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Quality of whole database:

GLP and guideline study.

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:

2004-11-12 to 2005-02-03

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

GLP compliance:

yes

Test type:

fixed dose procedure

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: 8-9 weeks
- Weight at study initiation: 229 g (range from 209 to 249 g).
- Fasting period before study: no
- Housing: separately in type III Makrolon cages
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22 to 23 °C
- Humidity: 44 to 57 %
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

semiocclusive

Vehicle:

water

Remarks:

Aqua pro injection, Merck KGaA, Darmstadt; Batch 189.

Details on dermal exposure:

Preparation: Before application, the test material was mixed with some drops of Aqua pro injectione and ground in a mortar using pestle. These test material were spread on the shaven skin.

Duration of exposure:

24 h

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5m / 5f

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: for at least 6 hours after administration and then checked daily
- Body weight: on days 2, 4, 6, 8, 11, 13, and 15
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other: gross pathology

Statistics:

The body weight data were recorded with the PC-program “AKUDAT“ The statistical evaluations of the body weight were carried out with the PC-program “TOX 511 A“, developed by the Institute of Toxicology of Merck KGaA, Darmstadt. The body weight development of each rat and group was determined. The group mean value was calculated for each measurement and printed on tables.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

Male: 2000 mg/kg bw; Number of animals: 5; Number of deaths: 0
Female: 2000 mg/kg bw; Number of animals: 5; Number of deaths: 0

Clinical signs:

other: Signs of toxicity related to dose levels: No signs of toxicity were detected @2000 mg/kg bw.

Gross pathology:

Effects on organs:
At necrospy, no organ alterations were seen.

Table 1 Summary of mortalities

Sex	LD50 on days 1 + 8+ 15
M + F	>2000 mg/kg bw

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the result of this study, the test item can be considered to have no acute toxic potential and to have a LD50 value higher than 2000 mg/kg bw after dermal application to rats.

Executive summary:

The study was performed accorting OECD TG 402. The test was performed as a limit test using a dose of 2000 mg/kg body weight. Directly before administration the test material was moistened and spread on the shaven skin in an area of 6 x 6 cm, and covered with a gauze patch. This was kept in place by a self-adhesive fabric (Fixomull® stretch, Beiersdorf). The time of exposure was 24 hours. Then the gauze and adhesive fabric were removed and any remaining test material was wiped off carefully. No signs of toxicity were detected in the rats (5 males and 5 females) after treatment with 2000 mg/kg bw. There were no deaths during the course of the study. The gross pathological examination revealed no organ alterations.

Based on the result of this study, the test material can be considered to have no acute toxic potential and to have a LD50 value higher than 2000 mg/kg bw after dermal application to rats.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

2 000 mg/kg bw

Quality of whole database:

GLP and guideline study.

Additional information

Acute oral toxicity

The acute toxicity of the test material was determined in rats after oral administration of 300 and 2000 mg/kg body weight. Directly before administration the test material was prepared with aqueous Methocel® K4M Premium solution as the vehicle. This study was performed according to the "Acute toxic class method" (ATC) as described in the OECD Guideline 423. After oral treatment with 300 mg/kg (3 males, 3 females), signs of toxicity were seen in the male and female rats. They started 1 to 2 hours after dosing and lasted up to 24 hours. They consisted of locomotor disturbance and dyspnea. All these animals survived the observation period. All rats, which were sacrificed at the end of the study, showed no macroscopic abnormalities.
After oral treatment with 2000 mg/kg (3 females), signs of toxicity were seen in the female rats. They started 1 to 15 minutes after administration and lasted up to 24 hours. They consisted of incomplete eyelid closure, abdominal position, respiratory sounds, locomotor disturbance, and dyspnea. One animal died 6 hours after administration. The two other animals were sacrificed in a moribund state 8 hours after administration. The female rat (2000 mg/kg), which died, showed changes in the gastrointestinal tract (liquid contents of the stomach and thickened contents of the cecum). The other two rats, which were sacrificed in moribund state showed no macroscopic abnormalities. According to the results of this study, the LD50 value of the test material is expected to be between 300 - 2000 mg/kg bw.

Acute dermal toxicity

The study was performed accorting OECD TG 402. The test was performed as a limit test using a dose of 2000 mg/kg body weight. Directly before administration the test material was moistened and spread on the shaven skin in an area of 6 x 6 cm, and covered with a gauze patch. This was kept in place by a self-adhesive fabric (Fixomull® stretch, Beiersdorf). The time of exposure was 24 hours. Then the gauze and adhesive fabric were removed and any remaining test material was wiped off carefully. No signs of toxicity were detected in the rats (5 males and 5 females) after treatment with 2000 mg/kg bw. There were no deaths during the course of the study. The gross pathological examination revealed no organ alterations. Based on the result of this study, the test material can be considered to have no acute toxic potential and to have a LD50 value higher than 2000 mg/kg bw after dermal application to rats.

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 classified and labelled as acutely toxic via the oral route Cat. 4 (H302 "harmful if swallowed") according to Regulation (EC) No 1272/2008 (CLP), as amended for the twelfth time in Regulation (EU) 2019/521.

Classification for dermal acute toxicity is not warranted.