Dipotassium dihydrogen 4-[5-[3-carboxylato-5-hydroxy-1-(4-sulphonatophenyl)-1H-pyrazol-4-yl]penta-2,4-dienylidene]-4,5-dihydro-5-oxo-1-(4-sulphonatophenyl)-1H-pyrazole-3-carboxylate

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

13 Feb - 13 Mar 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

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 2001

Deviations:

yes

Remarks:

/ no confirmatory step was performed with starting dose of 300 mg/kg bw; instead steps 2 and 3 were performed at 2000 mg/kg/day

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

adopted in 2008

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Version / remarks:

adopted in 2002

GLP compliance:

yes (incl. QA statement)

Remarks:

GLP-Landesleitstelle Bayern, Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, Schwabach, Germany

Test type:

acute toxic class method

Test material

Test material information

Constituent 1

Reference substance name:

Dipotassium dihydrogen 4-[5-[3-carboxylato-5-hydroxy-1-(4-sulphonatophenyl)-1H-pyrazol-4-yl]penta-2,4-dienylidene]-4,5-dihydro-5-oxo-1-(4-sulphonatophenyl)-1H-pyrazole-3-carboxylate

EC Number:

257-479-6

EC Name:

Dipotassium dihydrogen 4-[5-[3-carboxylato-5-hydroxy-1-(4-sulphonatophenyl)-1H-pyrazol-4-yl]penta-2,4-dienylidene]-4,5-dihydro-5-oxo-1-(4-sulphonatophenyl)-1H-pyrazole-3-carboxylate

Cas Number:

51858-17-4

Molecular formula:

C25H16N4O12S2.2K

IUPAC Name:

dipotassium dihydrogen 4-{5-[3-carboxylato-5-oxo-1-(4-sulfonatophenyl)-1,5-dihydro-4H-pyrazol-4-ylidene]penta-1,3-dien-1-yl}-5-hydroxy-1-(4-sulfonatophenyl)-1H-pyrazole-3-carboxylate

Test material form:

solid: crystalline

Test animals

Species:

rat

Strain:

Wistar

Remarks:

Crl: WI(Han)

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River, Sulzfeld, Germany
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 9 - 12 weeks
- Weight at study initiation: step 1: 168 - 187 g; step 2: 180 - 200 g; step 3: 191 - 196 g
- Fasting period before study: yes (food was withheld for 16 to 19 hours; access to water was permitted)
- Housing: group housing in in IVC cages, type III H, polysulphone cages on Altromin saw fibre bedding
- Diet: Altromin 1324 maintenance diet for rats and mice; ad libitum
- Water: tap water, sulphur-acidified to a pH value of approximately 2.8; ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 55 ± 10
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 14 Feb 2018 To: 13 Mar 2018

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

(aqua ad injectionem)

Details on oral exposure:

VEHICLE
- Concentration in vehicle: step 1: 0.3 g in 10 mL, steps 2 and 3: 2.0 g in 10 mL
- Amount of vehicle: 10 mL/kg bw

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

Doses:

300 and 2000 mg/kg bw

No. of animals per sex per dose:

300 mg/kg bw: 3 animals; 2000 mg/kg bw: 2 x 3 animals

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical examinations were made several times on the day of dosing (at least once during the first 30 minutes and with special attention given during the first 4 hours post-dose); thereafter, the animals were observed for clinical signs once daily until the end of the observation period. Body weights were recorded on day 1 (prior to the administration) and on days 8 and 15.
- Necropsy of survivors performed: yes

Statistics:

not applicable

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortalities were noted.

Clinical signs:

other: None of the animals treated at 300 mg/kg showed any signs of toxicity. At 2000 mg/kg bw all animals showed slight piloerection on the day of dosing. Further findings were blue discolored ears, snout, tail and eyes and extremities; additionally faeces and

Gross pathology:

No specific gross pathological changes were recorded for any animal.

Any other information on results incl. tables

Table 1. Absolute Body Weights in g and Body Weight Change in %

Step	Female No.	Dose	Body weight (g)			Body weight change in comparison to day 1 (%)
1	1	300	187	219	235	+26
	2		175	205	218	+25
	3		168	190	234	+39
2	4	2000	180	194	202	+12
	5		200	229	230	+15
	6		186	210	210	+13
3	7	2000	191	212	217	+14
	8		194	218	217	+12
	9		196	225	228	+16

Applicant's summary and conclusion

Interpretation of results:

other: CLP/GHS criteria not met; no classification required according to Regulation (EC) No. 1272/2008

Conclusions:

CLP: not classified
In this study in female rats an LD50 of > 2000 mg/kg bw was derived.