Registration Dossier

Administrative data

Endpoint:
methaemoglobinaemia
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Start of experiment: 12 March 1981 - End of experiment: 13 March 1981
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Pre GLP study, conducted according to sound scientific principles

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1981
Report date:
1981

Materials and methods

Test guideline
Qualifier:
no guideline available
GLP compliance:
no
Type of method:
in vivo
Endpoint addressed:
other: To investigate the Methaemoglobin-inducing capacity of the substance

Test material

Constituent 1
Chemical structure
Reference substance name:
Hexasodium 2,2'-[vinylenebis[(3-sulphonato-4,1-phenylene)imino[6-morpholino-1,3,5-triazine-4,2-diyl]imino]]bis(benzene-1,4-disulphonate)
EC Number:
257-827-7
EC Name:
Hexasodium 2,2'-[vinylenebis[(3-sulphonato-4,1-phenylene)imino[6-morpholino-1,3,5-triazine-4,2-diyl]imino]]bis(benzene-1,4-disulphonate)
Cas Number:
52301-70-9
Molecular formula:
C40H40N12O20S6.6Na
IUPAC Name:
hexasodium 2-{[4-({4-[(E)-2-[4-({4-[(2,5-disulfophenyl)amino]-6-(morpholin-4-yl)-1,3,5-triazin-2-yl}amino)-2-sulfophenyl]ethenyl]-3-sulfophenyl}amino)-6-(morpholin-4-yl)-1,3,5-triazin-2-yl]amino}benzene-1,4-disulfonate

Test animals

Species:
rat
Strain:
other: Tif: RAIf (SPF) strain
Sex:
male/female
Details on test animals or test system and environmental conditions:
Young adult rats of both sexes (Tif: RAIf (SPF) strain) weighing 200-250 g, bred and raised on the premises were used in the experiment. The rats were identified by individual tail and cage marking. They were maintained in groups of two in Macrolon Cages, Type 3 (Ehret, 783 Emmendingen / Germany) with wire mesh tops and saw dust bedding (granular form).
The animal room had a temperature of 22 ± 2 °C with 55 ± 10 % relative humidity and a 10 hour/day light cycle. Rat chow No. 890 (NAFAG, Gossau SG) and water were available ad libitum. The adaptation period was at least 7 days.

Test Material and Formulation:
The substance was dissolved in tap water in order to achieve a dosage level of 20 ml/kg body weight. Prior to the administration the formulation was homogeneously dispersed with an Ultra-Turrax and subsequently placed on a magnetic stirrer.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
Prior to the treatment the animals were fasted overnight. The formulated test compound was administered orally by gavage at a dosage of 5000 mg/kg. Symptoms and mortality were assessed 1, 2, 4, 6 and 24 hours after the administration.
Analytical verification of doses or concentrations:
no
Duration of treatment / exposure:
24 hours
Frequency of treatment:
single application
Doses / concentrations
Remarks:
Doses / Concentrations:
5000 mg/kg
Basis:

No. of animals per sex per dose:
16 animals in the control group
Control animals:
yes, concurrent vehicle

Examinations

Examinations:
Haematological investigations:
Blood sampling: Blood was collected by puncture of the retroorbital plexus using heparlnized micro-haematocrit tubes at the following times:
prior to the administration (all animals);
- 15, 30, 60, 90 minutes, 2, 4, 6 and 24 hours (each time hal f group).
The blood samples were immediately cooled and the haematological investigation was performed without delay.

Methods used in Haematology:
Haemoglobin and Methaemogl obin was determined with a IL 282 C0-0XYMETER and expressed as percentage of Haemoglobin.
Positive control:
The identical test as described above was performed with a negative control group of 16 rats, which received 20 ml/kg of tap water and a positive control group which received 40 mg/kg sodium nitrite solved in tap water, administered at a rate of 20 ml/kg.

Results and discussion

Details on results:
One animal died immediately after the administration of FAT 65037/D. Clinical signs of including sedation, dyspnoea, ruffled fur, diarrhoea and curved body position from moderate to severe were noted in all animals.

Applicant's summary and conclusion

Conclusions:
The substance has been found to be devoid of Methaemoglobin-inducing capacity after single oral administration of 5000 mg/kg in young adult rats.
Executive summary:

The purpose of the present study was to Methaemoglobin-inducing capacity of the substance comparison with a negative control (tap control group (sodium nitrite). The substance has been found to be devoid of Methaemoglobin-inducing capacity after single oral administration of 5000 mg/kg in young adult rats.