3,10-diamino-6,13-dichloro-2-((6-(((4-(1,1-dimethylethyl)phenyl)sulfonyl)amino)-2-naphthalenyl)sulfonyl)-4,11-triphenodioxazinedisulfonic acid, lithium potassium sodium salt

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

25 Sept 2001 - 18 Oct 2001

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2001

Report date:

2001

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

adopted March 22,1996

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

September 30, 1996

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

440-770-9

EC Name:

-

Cas Number:

371921-63-0

Molecular formula:

C38 H29 Cl2 N5 O12 S4 .x K .x Li .x Na

IUPAC Name:

3,10-diamino-2-{[6-(4-tert-butylbenzenesulfonamido)naphthalen-2-yl]sulfonyl}-6,13-dichloro-5,12-dioxa-7,14-diazapentacene-4,11-disulfonic acid lithium hydride potassium hydride sodium hydride

Test material form:

solid

Test animals

Species:

rat

Strain:

other: HanBrl: Wist (SPF)

Remarks:

Recognized by the international guidelines as a recommended test system.

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST SYSTEM
Source: RCC Ltd; Biotechnology and Animal Breeding Division; CH-4414 Füllinsdod / Switzerland
Number of animals per group: 3 males or 3 females
Total number of animals: 3 males and 3 females
Age when treated: Males: 8 weeks; Females: 10 weeks
ldentification: Unique cage card and corresponding color-coded spots on the tail.
Acclimatization: Under laboratory conditions, after health examination. Only animals without any visible signs of illness were used for the study.

HUSBANDRY
Room number: E16
Conditions: Standard Laboratory Conditions. Air-conditioned with 10-15 air changes per hour, and continuously monitored environment with target ranges for temperature 22 + 3"C and for relative humidity between 30-70%. 12 hours fluorescent light/12 hours
dark (light period between 6.00 and 18.00), music during the light period.

Accommodation: ln groups of three per sex in Makrolon type-4 cages with wire mesh tops and standardized softwood bedding ('Lignocel' Schill AG, CH-4132 Muttenz/Switzerland).

Diet: Pelleted standard Provimi Kliba 3433 raVmouse maintenance diet, batch no.73/01 (Provimi Kliba AG, CH-4303 Kaiseraugst Switzerland) ad libitum. Results of analyses for contaminants are archived at RCC Ltd, Itingen.

Water: Community tap-water, from ltingen ad libitum. Results of bacteriological, chemical and contaminant analyses are archived at RCC Ltd, ltingen

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

polyethylene glycol

Remarks:

PEG 300 from Fluka Chemie AG, CH-9471 Buchs (batch number: 42471811 42701)

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 0.2g test item / ml vehicle
- Amount of vehicle (if gavage): 10 ml vehicle/kg bw
- Justification for choice of vehicle: The vehicle was chosen after a non-GLP solubility trial performed before experimental starting date.
- Lot/batch no. (if required): Supplier: Fluka Chemie AG, CH-9471 Buchs (Batch: 42471811 42701)
- Purity: not specified

MAXIMUM DOSE VOLUME APPLIED: 2000 mg test item/kg bw

DOSAGE PREPARATION (if unusual): The preparations were made shortly before each dosing. The test item was weighed into a tared glass beaker on a suitable precision balance and the vehicle added (weight:volume). The mixtures were prepared using a magnetic stirrer. Homogeneity of the test item in the vehicle was maintained during administration using a magnetic stirrer.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: limit concentration of 2000 mg/kg bw

TREATMENT
The animals received a single oral dose of the test item by gavage at 2000 mg/kg body weight after being fasted for 16 to 20 hours (access to water was permitted). Food was provided again approximately 3 hours after dosing.

Doses:

0: 2000 mg/kg bw

No. of animals per sex per dose:

3

Control animals:

yes

Details on study design:

- Duration of observation period following administration: 15 days
- Frequency of observations and weighing:
Mortality / Viability Daily during acclimatization and twice daily during days 1-15.
Body weights On test days 1 (pre-administration), I and 15.
- Necropsy of survivors performed: yes
All surviving animals were killed at the end of the observation period by an intraperitoneal injection of NARCOREN (Rhône Mérieux GmbH, D-88471 Laupheim) at a dose of at least
2.0 ml/kg body weight (equivalent to at least 320 mg sodium pentobarbitone/kg body weight) and discarded after macroscopic examinations were pedormed. No organs or tissues were retained.
- Other examinations performed: clinical signs: Daily during acclimatization and at least four times on test day 1 after the test item administration. Once daily during days 2-15. All abnormalities were recorded.

Statistics:

No statistical analysis was used.

Results and discussion

Preliminary study:

Not performed

Effect levels

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No death occurred during the study.

Clinical signs:

No clinical signs were noted during the course of the study.

Body weight:

The body weight of the animals was within the range commonly recorded for this strain and age.

Gross pathology:

No macroscopic findings were observed at necropsy.

Any other information on results incl. tables

Body weight evolution

Dose [mg/kg]	Animal No.	Sex	Body weight [g]
			Day 1 (treatment)	Day 8	Day 15
2000	1	F	181 .6	200.7	209.2
2000	2	F	178.1	199.3	209.1
2000	3	F	178.7	197.4	207.7
2000	4	M	195.8	243.0	270.6
2000	5	M	208.0	261.7	284.9
2000	6	M	204.7	257.2	284.9

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The oral LDso (rat) of BLUE GS 5664.80 is > 2000 mg/kg bw in test performed according to OECD TG 423 and following GLP.

Executive summary:

The acute oral toxicity of BLUE GS 5664.80 has been determined in the current study performed following OECD TG 423 according to GLP.

One group of three male or three female HanBrl: WIST (SPF) rats was treated by oral gavage with BLUE GS 5664.80 at 2000 mg/kg body weight. The test item was suspended in vehicle (PEG 300) at a concentration of 0.2 g/ml and administered at a volume of 10 ml/kg.

The animals were examined for clinical signs daily during the acclimatization period, four times during test day 1 and once daily during test days 2-15. Mortality/viability was recorded daily during the acclimatization period and together with clinical signs at the same time intervals on test day 1 and twice daily on test days 2-15. Body weights were recorded on day 1 (priorto administration) and on days 1 and 15. All animals were necropsied and examined macroscopically.

All animals survived until the end of the study period.

No clinical signs were evident during the course of the study.

The body weight of the animals was within the range commonly recorded for this strain and age.

No macroscopic findings were observed at necropsy.

The median lethal dose of BLUE GS 5664.80 after single oral administration to rats of both sexes, observed over a period of 14 days is:

LDso (rat): > 2000 mg/kg bw