Cuprate(4-), [C-(aminosulfonyl)-C-[[[2-[[4-chloro-6-[(2,5-disulfophenyl)amino]-1,3,5-triazin-2-yl]amino]ethyl]amino]sulfonyl]-29H,31H-phthalocyanine-C,C-disulfonato(6-)-N29,N30,N31,N32]-, sodium salts

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

27 April 1993 to 8 June 1993.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1993

Report date:

1993

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Version / remarks:

OECD Guidelines for Testing of Chemicals (1987) No. 401 "Acute Oral Toxicity"

Deviations:

not specified

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.1 (Acute Toxicity (Oral))

Version / remarks:

Method B1 in Commission Directive 92/69/EEC (which constitutes Annex V of Council Directive 67/548/EEC).

Deviations:

not specified

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Cuprate(4-), [C-(aminosulfonyl)-C-[[[2-[[4-chloro-6-[(2,5-disulfophenyl)amino]-1,3,5-triazin-2-yl]amino]ethyl]amino]sulfonyl]-29H,31H-phthalocyanine-C,C-disulfonato(6-)-N29,N30,N31,N32]-, sodium salts

EC Number:

689-188-9

Cas Number:

149343-84-0

Molecular formula:

C32H24N12O8S4Cu to C76H56N28O32S12Cl4Na8Cu

IUPAC Name:

Cuprate(4-), [C-(aminosulfonyl)-C-[[[2-[[4-chloro-6-[(2,5-disulfophenyl)amino]-1,3,5-triazin-2-yl]amino]ethyl]amino]sulfonyl]-29H,31H-phthalocyanine-C,C-disulfonato(6-)-N29,N30,N31,N32]-, sodium salts

Test material form:

solid: particulate/powder

Details on test material:

Identification: JPR Blue 100
Description: Blue powder
Chemical Name: Cuprate(2-),[2-[[4-chloro-6-[[2-[(29H,31H-phthalocyaninylsulphonyl)amino]ethyl]amino]-1,3,5-triazin-2-yl]amino]-1,4-benzenedisulphonato(4-)-N29,N30,N31,N32]-mono(or bis)aminosulphonyl mono (or di)sulpho derivs., sodium salts.
Lot Number: 303001
Purity: 95.2%
Major Impurities: H2o, NaCl, Na2SO4
Date Received: 30 March 1993
Container: Opaque plastic jar x 7
Storage conditions: Room temperature

Specific details on test material used for the study:

No further details specified in the study report.

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

Specification
Male and female Sprague-Dawley strain rats were supplied by Charles River (UK) Ltd., Manston, Kent, U.K. At the start of the main study the males weighed 138 – 165 g, and the females 122 – 134 g, and were approximately five to eight weeks old. After a minimum acclimatization period of at least five days the animals were selected at random and given a unique number within the study by indelible ink marking on the tail and a number written on a cage card.

Husbandry
The animals were housed in groups of five by sex in solid-floor polypropylene cages with sawdust bedding. With the exception of an overnight fast immediately before dosing and for approximately two hours after dosing, free access to mains drinking water and food (Rat and Mouse Expanded Diet No. 1, Special Diet Services Limited, Witham, Essex, U.K.) was allowed throughout the study.
The animal room was maintained at a temperature of 19 – 22 °C and relative humidity of 48 - 54%. The rate of air exchange was approximately 15 changes per hour and the lighting was controlled by a time switch to give 12 hours continuous light and 12 hours darkness.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

For the purpose of the study the test material was freshly prepared, as required, as a suspension at the appropriate concentration in distilled water.

Doses:

Single dose: 2000 mg/kg

No. of animals per sex per dose:

Range-finding study: 2 (1 male/1 female)
Main Study: 10 (5 males/5 females)

Control animals:

no

Details on study design:

Range-finding Study
A range-finding study was performed to establish a dosing regime.
The animals were observed for deaths or overt signs of toxicity ½, 1, 2 and 4 hours after dosing and subsequently once daily for 5 days.
Individual bodyweights were recorded on the day of dosing to allow calculation of individual treatment volumes. No necropsies were performed.

Main Study
Based on the results of the range-finding study a further group of animals was treated.
All animals were dosed once only by gavage using a metal cannula attached to a graduated syringe. The volume administered to each animal was calculated according to its fasted bodyweight at the time of dosing.
The animals were observed for deaths or overt signs of toxicity ½, 1, 2 and 4 hours after dosing and subsequently once daily for 14 days.
Individual bodyweights were recorded prior to dosing on Day 0 and on Days 7 and 14.
At the end of the study the animals were killed by cervical dislocation and subjected to gross pathological examination. This consisted of an external examination and opening of the abdominal and thoracic cavities. The appearance of any macroscopic abnormalities was recorded. No tissues were retained.

Results and discussion

Preliminary study:

There were no deaths or clinical signs of toxicity.
Based on this information, a dose level of 2000 mg/kg bodyweight was selected for the main study.

Effect levels

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

There were no deaths.

Clinical signs:

Blue-coloured urine was noted in all animals. Blue-coloured staining of the fur was noted in all animals. All animals recovered two days after dosing .

Body weight:

All animals showed expected gain in bodyweight during the study.

Gross pathology:

All animals showed dark blue-coloured outer cortex of the kidneys at necropsy. The females also showed blue-coloured staining of the non-glandular epithelium of the stomach.

Any other information on results incl. tables

JPR BLUE 100: ACUTE ORAL TOXICITY (LIMIT TEST) IN THE RAT

INDIVIDUAL CLINICAL OBSERVATIONS AND MORTALITY DATA IN THE RANGE-FINDING STUDY

Dose Level mg/kg	Animal Number & Sex	Effects Noted After Dosing (Hours)				Effects Noted During Period After Dosing (Days)
		½	1	2	4	1	2	3	4	5
2000	1-0 Male	0	0	0	0	0	0	0	0	0
	2-0 Female	0	0	0	0	0	0	0	0	0

 

JPR BLUE 100: ACUTE ORAL TOXICITY (LIMIT TEST) IN THE RAT

INDIVIDUAL CLINICAL OBSERVATIONS AND MORTALITY DATA IN THE MAIN STUDY

Dose Level mg/kg	Animal Number & Sex	Effects Noted After Dosing (Hours)				Effects Noted During Period After Dosing (Days)
		½	1	2	4	1	2	3	4	5	6	7	8	9	10	11	12	13	14
2000	3-0 Male	0	0	0	0	Fs DuB	Fs0	0	0	0	0	0	0	0	0	0	0	0	0
	3-1 Male	0	0	0	0	Fs DuB	Fs0	0	0	0	0	0	0	0	0	0	0	0	0
	3-2 Male	0	0	Fs0	Fs0	Fs DuB	Fs0	0	0	0	0	0	0	0	0	0	0	0	0
	3-3 Male	0	0	0	0	Fs DuB	Fs0	0	0	0	0	0	0	0	0	0	0	0	0
	3-4 Male	0	0	0	0	Fs DuB	Fs0	0	0	0	0	0	0	0	0	0	0	0	0
	4-0 Female	0	0	Fs0	Fs0	Fs DuB	Fs0	0	0	0	0	0	0	0	0	0	0	0	0
	4-1 Female	0	0	0	Fs0	Fs DuB	Fs0	0	0	0	0	0	0	0	0	0	0	0	0
	4-2 Female	0	0	0	0	Fs DuB	Fs0	0	0	0	0	0	0	0	0	0	0	0	0
	4-3 Female	0	0	Fs0	Fs DuB	Fs DuB	Fs0	0	0	0	0	0	0	0	0	0	0	0	0
	4-4 Female	0	0	0	0	Fs DuB	Fs0	0	0	0	0	0	0	0	0	0	0	0	0

0 = no signs of systemic toxicity                                                      DuB = blue coloured urine

Fs = blue coloured staining of the fur

 

JPR BLUE 100: ACUTE ORAL TOXICITY (LIMIT TEST) IN THE RAT

INDIVIDUAL BODYWEIGHTS AND WEEKLY BODYWEIGHT GAIN IN THE MAIN STUDY

Dose Level mg/kg	Animal Number & Sex	Bodyweight (g) at Day			Bodyweight Gain (g) During Week
		0	7	14	1	2
2000	3-0 Male 3-1 Male 3-2 Male 3-3 Male 3-4 Male 4-0 Female 4-1 Female 4-2 Female 4-3 Female 4-4 Female	161 150 165 138 164 122 132 130 134 128	226 238 240 202 287 178 190 179 175 171	287 290 308 254 295 206 221 203 195 188	65 88 75 64 73 56 58 49 41 43	61 52 68 52 58 28 31 24 20 17

 

JPR BLUE 100: ACUTE ORAL TOXICITY (LIMIT TEST) IN THE RAT

INDIVIDUAL NECROPSY FINDINGS IN THE MAIN STUDY

Dose Level mg/kg	Animal Number & Sex	Time of Death	Macroscopic Observations
2000	3-0 Male	Killed day 14	Kidneys: dark blue coloured outer cortex region
	3-1 Male	Killed day 14	Kidneys: dark blue coloured outer cortex region
	3-2 Male	Killed day 14	Kidneys: dark blue coloured outer cortex region
	3-3 Male	Killed day 14	Kidneys: dark blue coloured outer cortex region
	3-4 Male	Killed day 14	Kidneys: dark blue coloured outer cortex region
	4-0 Female	Killed day 14	Kidneys: dark blue coloured outer cortex region Non-glandular epithelium of stomach: blue coloured staining
	4-1 Female	Killed day 14	Kidneys: dark blue coloured outer cortex region Non-glandular epithelium of stomach: blue coloured staining
	4-2 Female	Killed day 14	Kidneys: dark blue coloured outer cortex region Non-glandular epithelium of stomach: blue coloured staining
	4-3 Female	Killed day 14	Kidneys: dark blue coloured outer cortex region Non-glandular epithelium of stomach: blue coloured staining
	4-4 Female	Killed day 14	Kidneys: dark blue coloured outer cortex region Non-glandular epithelium of stomach: blue coloured staining

 

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The acute oral median lethal dose (LDso) of the test material, JPR BLUE 100, in the Sprague-Dawley strain rat was found to be greater than 2000 mg/kg bodyweight. No symbol and risk phrase are required according to EEC labelling regulations.

Executive summary:

A study was performed to assess the acute oral toxicity of the test material in the Sprague-Dawley strain rat. The method used followed that described in the OECD Guidelines for Testing of Chemicals (1987) No. 401 "Acute Oral Toxicity" referenced as Method B1 in Commission Directive 92/ 69/ EEC (which constitutes Annex V of Council Directive 67/ 548/ EEC).

 

The results may be used as a basis for classification and labelling under Annex VI of Council Directive 67/ 548/ EEC (as adapted to technical progress by Commission Directive 91/325/ EEC).

 

Following a range-finding study, a group of ten fasted animals (five males and five females) was given a single oral dose of test material, as a suspension in distilled water at a dose level of 2000 mg/kg bodyweight. The animals were observed for fourteen days after the day of dosing and were then killed for gross pathological examination.

 

There were no deaths. Blue-coloured urine was noted in all animals.

All animals recovered two days after dosing.

 

All animals showed expected gain in bodyweight during the study.

All animals showed dark blue-coloured outer cortex of the kidneys at necropsy. The females also showed blue-coloured staining of the non-glandular epithelium of the stomach.

 

The acute oral median lethal dose (LD50) of the test material in the Sprague -Dawley strain rat was found to be greater than 2000 mg/kg bodyweight. No symbol and risk phrase are required according to EEC labelling regulations.