Reaction product of 9-[2-(ethoxycarbonyl)phenyl]-3,6-bis(ethylamino)-2,7-dimethylxanthylium chloride and Naphthalenesulfonic acids, reaction products with formaldehyde, ammonium salts

Endpoint summary

Description of key information

The test item induced mortality following a single oral administration to female rats at a dose of 2000 and 300 mg/kg bw and did not induced mortality at a dose of 50 mg/kg bw. The LD50 was determined to be between 50 and 300 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:

2013-10-29 to 2013-11-20

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study according to OECD/EU guideline.

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

17 December 2001

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

30 May 2008

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Version / remarks:

December 2002

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class 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.
- Age at study initiation: Young adult rats, 11 weeks old in group 1, 2 and 3 and 12 weeks old in group 4 and 5
- Weight at study initiation: 190-220 g
- Fasting period before study: yes
- Housing: 3 animals/sex/cage
- Diet: ssniff® SM R/M-Z+H complete diet ad libitum
- Water: tap water ad libitum
- Acclimation period: 26 to 33 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 30 - 70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): Artificial light, from 6 a.m. to 6 p.m.

Route of administration:

oral: gavage

Vehicle:

other: 1 % Methylcellulose solution

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 200, 30 and 5 mg/mL
- Amount of vehicle: 10 mL/kg bw
- Lot/batch no.: N83746634

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 on three female rats.

Doses:

First step: 2000 mg/kg bw
Second step: 300 mg/kg bw
Third step: 50 mg/kg bw

No. of animals per sex per dose:

3 per treatment or 6 animals per dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations: 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 each day for 14 days thereafter.
- Frequency of weighing: The body weight were recorded on day 0 (shortly before the treatment), on day 7, and on day 15
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight

Statistics:

not applicable

Sex:

female

Dose descriptor:

LD50

Effect level:

> 50 - < 300 mg/kg bw

Based on:

test mat.

Remarks on result:

other: 0/6 animals died at 50 mg/kg bw.

Mortality:

Two rats dosed at 2000 mg/kg bw DOCU Red 116 died on Day 1. Two of six animals treated with 300 mg/kg bw dose of test item died during the study. One animal of group 3 died on Day 3 and one animal of group 3 died on Day 2. All deaths seemed to be consequences of systemic toxic effect of the test item.
No death occurred at 50 mg/kg single oral dose of the test item. All rats in step 4 and step 5 survived until the end of the 14-day observation period.

Clinical signs:

other: Please refer to "Other findings"

Gross pathology:

Two rats treated with 2000 mg/kg bw dose of the test item spontaneously died during the study. One animal treated with the same dose survived until the scheduled necropsy on Day 15. An internal necropsy finding as autolysis was observed in two animals. It is normal physiological process after death. Besides, internal necropsy finding as pale kidneys was observed in one animal of group 1. This alteration could not be related to the test item toxic effect, but was regarded an individual variation. Most likely the observation is a congenital anomaly. Moderate hydrometra was observed in one female of the group 1 is physiological finding and connected to the oestrus cycle of the animal.
Two rats treated with 300 mg/kg bw dose of the test item spontaneously died during the study and four animals treated with same dose survived until the scheduled necropsy on Day 15. An internal necropsy finding as autolysis was observed in two animals of group 3. It is normal physiological process after death. Besides, internal necropsy finding as pale kidneys was observed in one animal of group 3 and in one animal of group 2. This alteration could not be related to the test item toxic effect, but was regarded an individual variation. Most likely the observation is a congenital anomaly. Moderate hydrometra was observed in two females of the group 2 and in one animal of group 3 is physiological finding and connected to the oestrus cycle of the animal.
All animals treated with 50 mg/kg bw dose of test item survived until the scheduled necropsy on Day 15. No pathological changes related to the effect of the test item were found during the macroscopic examination of animals.

Other findings:

In group 1, treated with 2000 mg/kg bw dose, clinical signs of reaction comprised of decreased activity (14 cases of 29 observations), abnormal gait (13/29), crouching (13/29), red coloured faces (15/29) and piloerection (15/29). Decreased activity, abnormal gait, crouching, red coloured faces and piloerection were observed in all animals. These symptoms were detected between treatment day and Day 6. All animals were free of symptoms on the treatment day 30 min. and 1 hour after the treatment, as well as the survivor animal was free of symptoms between Day 7 and Day 14.
In group 2, treated with 300 mg/kg bw dose, clinical signs of reaction comprised of blood around the nose (1 case of 57 observations), red coloured faces (24/57) and piloerection (5/57). Red coloured faces were observed in all animals. Blood around the nose and piloerection were recorded in one animal. These symptoms were detected between treatment day and Day 5. All animals were free of symptoms on the treatment day 30 min. and 1 hour after the treatment, as well as between Day 6 and Day 14.
In group 3, treated with 300 mg/kg bw dose, clinical signs of reaction comprised of decreased activity (14 cases of 32 observations), abnormal gait (11/32), crouching (12/32), red coloured faces (14/32) and piloerection (12/32). Decreased activity, abnormal gait, crouching, red coloured faces and piloerection were observed in all animals. These symptoms were detected between treatment day and Day 5. All animals were free of symptoms on the treatment day 30 min. and 1 hour after the treatment, as well as the survivor animal was free of symptoms on the treatment day 2 hours after the treatment and between Day 6 and Day 14.
In group 4, treated with 50 mg/kg bw dose, clinical signs of reaction comprised of red coloured faces (12 case of 57 observations) and piloerection (1/57). Red coloured faces were observed in all animals. Piloerection were recorded in one animal. These symptoms were detected between treatment day and Day 2. All animals were free of symptoms on the treatment day 30 min., 1 and 2 hours after the treatment, as well as between Day 3 and Day 14.
In group 5, treated with 50 mg/kg bw dose, clinical signs of reaction comprised of red coloured faces (12 cases of 57 observations). This symptom was observed in all animals on the treatment day 3 and 4 hours after the treatment and between Day 1 and Day 2.

Interpretation of results:

Toxicity Category III

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The test item induced mortality following a single oral administration to female rats at a dose of 2000 and 300 mg/kg bw and did not induced mortality at a dose of 50 mg/kg bw. The LD50 was determined to be between 50 and 300 mg/kg bw.

Executive summary:

The acute oral toxicity of the test item was assessed in an acute oral toxicity study (acute toxic class method) using Wistar rats according to OECD TG 423 (17 December 2001) and EU guideline B.1 tris (30 May 2008). Starting dose was selected on the basis of the available information about the test item.

Animals were weighed, observed for lethality and toxic symptoms for 14 days after the treatment. Gross pathological examination was carried out in animals died on Day 1, Day 2 and Day 3, as well as 15th day after the treatment in survivor animals.

Two of three animals treated with 2000 mg/kg bw DOCU Red 116 died on Day 1. Two of six animals treated with 300 mg/kg bw dose died on Day 2 and on Day 3, respectively. No lethality was noted at single oral dose of 50 mg/kg bw.

In the first step, CNS symptoms (decreased activity, crouching), disturbances of coordination (abnormal gait), disturbance of the autonomic functions (piloerection) and red coloured faces were observed in animals between the treatment day and Day 6 post-dose. In the second step, blood around the nose, disturbance of the autonomic functions (piloerection) and red coloured faces were observed in animals between the treatment day and Day 5 post-dose. In the third step, CNS symptoms (decreased activity, crouching), disturbances of coordination (abnormal gait), disturbance of the autonomic functions (piloerection) and red coloured faces were observed in animals between the treatment day and Day 5 post-dose. In the fourth step, disturbance of the autonomic functions (piloerection) and red coloured faces were observed in animals between the treatment day and Day 2 post-dose. In the fifth step, red coloured faces were observed in animals between the treatment day and Day 2 post-dose.

The body weight development was undisturbed in all survival animals.

Altogether 4 animals died, and 11 animals were sacrificed as scheduled during the study.

All organs of the animals proved to be free of treatment related gross pathological changes.

The test item induced mortality following a single oral administration to female rats at a dose of 2000 and 300 mg/kg bw and did not induced mortality at a dose of 50 mg/kg bw.

The clinical symptoms observed were connected with systemic toxic effect of the test item, except red coloured faces, which was connected with the physical property of test item.

No test item related effects on body weights and body weight gains were observable during the study. Autopsy revealed no treatment related pathological changes.

The method used is not intended for the precise calculation of a precise LD50 value.

The test item was ranked into acute toxicity, oral, category 3 according to the current EU Regulation on classification, labelling and packaging (CLP) (EC) No 1272/2008:

 

Dose (mg/kg bw)	Mortality (dead/treated)	LD 50 (mg/kg bw)	CLP category
50	0/6	between 50 and 300	3

 

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

discriminating dose

Value:

50 mg/kg bw

Quality of whole database:

GLP study according to OECD/EU guideline.

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Oral

The acute oral toxicity of the test item was assessed in an acute oral toxicity study (acute toxic class method) using Wistar rats according to OECD TG 423 (17 December 2001) and EU guideline B.1 tris (30 May 2008). Starting dose was selected on the basis of the available information about the test item.

Animals were weighed, observed for lethality and toxic symptoms for 14 days after the treatment. Gross pathological examination was carried out in animals died on Day 1, Day 2 and Day 3, as well as 15th day after the treatment in survivor animals.

Two of three animals treated with 2000 mg/kg bw DOCU Red 116 died on Day 1. Two of six animals treated with 300 mg/kg bw dose died on Day 2 and on Day 3, respectively. No lethality was noted at single oral dose of 50 mg/kg bw.

In the first step, CNS symptoms (decreased activity, crouching), disturbances of coordination (abnormal gait), disturbance of the autonomic functions (piloerection) and red coloured faces were observed in animals between the treatment day and Day 6 post-dose. In the second step, blood around the nose, disturbance of the autonomic functions (piloerection) and red coloured faces were observed in animals between the treatment day and Day 5 post-dose. In the third step, CNS symptoms (decreased activity, crouching), disturbances of coordination (abnormal gait), disturbance of the autonomic functions (piloerection) and red coloured faces were observed in animals between the treatment day and Day 5 post-dose. In the fourth step, disturbance of the autonomic functions (piloerection) and red coloured faces were observed in animals between the treatment day and Day 2 post-dose. In the fifth step, red coloured faces were observed in animals between the treatment day and Day 2 post-dose.

The body weight development was undisturbed in all survival animals.

Altogether 4 animals died, and 11 animals were sacrificed as scheduled during the study.

All organs of the animals proved to be free of treatment related gross pathological changes.

The test item induced mortality following a single oral administration to female rats at a dose of 2000 and 300 mg/kg bw and did not induced mortality at a dose of 50 mg/kg bw.

The clinical symptoms observed were connected with systemic toxic effect of the test item, except red coloured faces, which was connected with the physical property of test item.

No test item related effects on body weights and body weight gains were observable during the study. Autopsy revealed no treatment related pathological changes.

The method used is not intended for the precise calculation of a precise LD50 value.

The test item was ranked into acute toxicity, oral, category 3 according to the current EU Regulation on classification, labelling and packaging (CLP) (EC) No 1272/2008:

 

Dose (mg/kg bw)	Mortality (dead/treated)	LD 50 (mg/kg bw)	CLP category
50	0/6	between 50 and 300	3

 

Dermal and by inhalation

No study available.

Justification for selection of acute toxicity – oral endpoint
Only one study available.

Justification for classification or non-classification

Based on the available data on acute oral toxicity the test item is classified for acute oral toxicity H310, Toxic if swallowed according to Regulation (EC) No 1272/2008 (CLP) and T, R25 according to Directive 67/548/EEC (DSD).