Hydrogen [4-[4-(diethylamino)-2',4'-disulphonatobenzhydrylidene]cyclohexa-2,5-dien-1-ylidene]diethylammonium, sodium salt

Carcinogenicity

Description of key information

The Carcinogenic NOAEL value for the test substance hydrogen [4-[4-(diethylamino)-2', 4'-disulphonatobenzhydrylidene]cyclohexa-2,5-dien-1-ylidene]diethylammonium, sodium salt  is found to be 784.6 mg/kg bw. in rat. Thus hydrogen [4-[4-(diethylamino)-2', 4'-disulphonatobenzhydrylidene]cyclohexa-2,5-dien-1-ylidene]diethylammonium, sodium salt  is found to be non carcinogenc in nature.

Key value for chemical safety assessment

Carcinogenicity: via oral route

Link to relevant study records

Reference

Endpoint:

carcinogenicity

Remarks:

subcutaneous

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Data is from peer reviewed journal

Qualifier:

according to guideline

Guideline:

other: as below

Principles of method if other than guideline:

The sequential histological changes and neoplastic response occurring in the subcutaneous tissue of rats after injection of Patent Blue V was performed to determine its carcinogenic potential.

GLP compliance:

not specified

Species:

rat

Strain:

other: Ash/CFE

Sex:

male/female

Details on test animals or test system and environmental conditions:

Details on test animal and environmental conditions: TEST ANIMALS- Source: No data available- Age at study initiation: No data available- Weight at study initiation: average 100g- Fasting period before study: No data available- Housing: No data available- Diet (e.g. ad libitum): Spillers small laboratory animal diet, ad libitum- Water (e.g. ad libitum): Water, ad libitum- Acclimation period: No data availableENVIRONMENTAL CONDITIONS- Temperature (°C): 22 ± 1 °C - Humidity (%): 50%- Air changes (per hr): No data available- Photoperiod (hrs dark / hrs light): No data availableIN-LIFE DATES: From: To: No data available

Route of administration:

subcutaneous

Type of inhalation exposure (if applicable):

not specified

Vehicle:

other: Co2 free distilled water

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: Solutions were prepared using CO2 free distilled water. Colouring (Blue VRS) were filtered and buffered to pH 7.0 beforeand prepared freshly before injection in a volum of 0.5 mlDIET PREPARATION- Rate of preparation of diet (frequency): No data available- Mixing appropriate amounts with (Type of food): No data available- Storage temperature of food: No data availableVEHICLE- Justification for use and choice of vehicle (if other than water): CO2 free distilled water - Concentration in vehicle: 2%- Amount of vehicle (if gavage): No data available- Lot/batch no. (if required): No data available- Purity: No data available

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

60 weeks

Frequency of treatment:

Twice weekly

Post exposure period:

No data available

Remarks:

Doses / Concentrations:784.6 mg/Kg (2%)Basis:no data

No. of animals per sex per dose:

132 in total

Control animals:

yes, concurrent vehicle

Details on study design:

No data available

Positive control:

N-methyl-N-nitrosourea (MNU) and N-nitroquinoline-N-oxide (NQO) were used as positive control.

Observations and examinations performed and frequency:

Observation and examinations performed and frequency:CAGE SIDE OBSERVATIONS:Yes- Time schedule:- Cage side observations checked in table [No.?] were included.: Survival were observed. DETAILED CLINICAL OBSERVATIONS: Yes - Time schedule: throughout the study experimentDERMAL IRRITATION (if dermal study): No data available - Time schedule for examinations:BODY WEIGHT: No data available- Time schedule for examinations:FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): No data availableFOOD EFFICIENCY: No data availableOPHTHALMOSCOPIC EXAMINATION: No data available HAEMATOLOGY: No data availableCLINICAL CHEMISTRY: No data availableURINALYSIS: No data available OTHER: The subcutaneous injection was given in the right flank of each rat and the injection site was shaved with electric clippersTissue reactions to Blue VRS were studied in two ways—A) Rats injected throughout the experimental periodB) Rats injected for only part of the experimental period

Sacrifice and pathology:

GROSS PATHOLOGY: No data available HISTOPATHOLOGY: Yes

Other examinations:

No data available

Statistics:

No data available

Clinical signs:

not specified

Mortality:

not specified

Body weight and weight changes:

not specified

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

not specified

Histopathological findings: neoplastic:

no effects observed

Details on results:

Gross Pathology: No macroscopical abnormalities at the injection site were observed in treated male and female rats as compared to controls. Histopathology: neoplastic:No resolution was observed in any rat treated with 784.6 mg/kg that had received more than 25 injections, although areas of fatty tissue could be seen between the collagen strands in animals receiving less than 40 injections.In rats that had received more than 40 injections thick scar tissue containing wide collagen bands filled the subcutaneous area, and foci of fibroblastic activity were observed in several animals. Other animals had developed sarcomata locally.

Dose descriptor:

NOAEL

Effect level:

784.6 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No effect on survival and histopathology

Remarks on result:

other:

Remarks:

Effect type: other: Tumorogenic (migrated information)

Incidence of local tissue response elicited by surfactants in rats after increasing number of injections

 

Rats injected throughout the experimental period and killed 24 hrs after the last injection
No. of injections	Total rats	Quiescent scar tissue	Foci of fibroblastic activity	Sarcomata
1-24	12	0	6	0
32-52	14	4	10	0
56-76	12	1	10	1
80-96	11	1	8	2
100-117	11	0	3	8

 

Rats injected for only part of the experimental period and killed at termination of experiment or when tumors developed
No. of injections	Total rats	Quiescent scar tissue	Foci of fibroblastic activity	Sarcomata
1-28	15	0	0	0
32-60	15	9	5	1
64-92	15	5	4	6
96-117	17	4	10	3

 

Conclusions:

NOAEL was considered to be 78436 mg/kg when Ash/CFE male and female rats were treated with Blue VRS subcutaneously for 60 weeks.

Executive summary:

In a Tumorogenecity study, Ash/CFE male and female rats were treated with Blue VRS in the concentration of 2 % (784.6 mg/kg) subcutaneously as a Solutions prepared using CO₂ free distilled water. Colouring (Blue VRS) were filtered and buffered to pH 7.0 before injection. No macroscopical abnormalities at the injection site were observed in treated male and female rats as compared to controls. In addition, No resolution were observed in any rat treated with 784.6 mg/kg that had received more than 25 injections, although areas of fatty tissue could be seen between the collagen strands in animals receiving less than 40 injections. In rats that had received more than 40 injections thick scar tissue containing wide collagen bands filled the subcutaneous area, and foci of fibroblastic activity were observed in several animals. Other animals had developed sarcomata locally. Therefore, NOAEL was considered to be 78436 mg/kg when Ash/CFE male and female rats were treated with Blue VRS subcutaneously for 60 weeks.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

784.6 mg/kg bw/day

Study duration:

chronic

Species:

rat

Quality of whole database:

Data from K2 publication.

Carcinogenicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Carcinogenicity: via dermal route

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Carcinogenicity:

Based on available data for target Hydrogen [4-[4-(diethylamino)-2',4'-disulphonatobenzhydrylidene]cyclohexa-2,5-dien-1-ylidene]diethylammonium, sodium salt (CAS no 129-17-9) for carcinogenicity is summaries below

In a study conducted by Hoosonet al(1973), Tumorogenecity was evaluated in Ash/CFE male and female rats by using Blue VRS in the concentration of 2 % (784.6 mg/kg) subcutaneously as a Solutions prepared using CO₂free distilled water. Colouring (Blue VRS) were filtered and buffered to pH 7.0 before injection. No macroscopical abnormalities at the injection site were observed in treated male and female rats as compared to controls. In addition, No resolution were observed in any rat treated with 784.6 mg/kg that had received more than 25 injections, although areas of fatty tissue could be seen between the collagen strands in animals receiving less than 40 injections. In rats that had received more than 40 injections thick scar tissue containing wide collagen bands filled the subcutaneous area, and foci of fibroblastic activity were observed in several animals. Other animals had developed sarcomata locally. Therefore, NOAEL was considered to be 78436 mg/kg when Ash/CFE male and female rats were treated with Blue VRS subcutaneously for 60 weeks. 

In a study given by EFSA Panel (2013), combined repeated dose & carcinogenicity was evaluated in male and female mice by using Patent Blue Vin the concentration of0, 0.1, 0.3 and 1 % (equivalent to 0, 150, 500 and 1500 mg/kg body weight/day) orally. Slightly increased in mortality,Significant decrease in body weight of male mice andSignificant reduction in haemoglobin, haematocrit, red blood cell counts and reticulocyte count, increase in the total number of white blood cells inmale and female rat were observed at 1 % dose group as compared to control. Increased in relative heart and caecal weights were observed in female at 1 % dose group. In addition,Non-neoplastic lesions of lungs, kidneys and liver were observed. Adrenocortical adenoma and carcinoma, adenoma of mammary gland were observed but the observed changes were mainly isolated findings, of commonly occurring tumours showing no dose-response relationship. Therefore, NOAEL was considered to be 1500 mg/kg body weight /day when male and female mice were treated with Patent Blue V orally for approx. 32 weeks.

Thus, based on above available data forfor target Hydrogen [4-[4-(diethylamino)-2',4'-disulphonatobenzhydrylidene]cyclohexa-2,5-dien-1-ylidene]diethylammonium, sodium salt (CAS no 129-17-9) is likely to be non hazardous as carcinogen.

Justification for selection of carcinogenicity via oral route endpoint:

There was some evidence of carcinogenic activity in male and female Ash/CFE rats based Tumorogenic effect of the test animals.

The carcinogenic Low Observed Adverse Effect Level (NOAEL) for the given test material is found to be 1000 mg/kg bw.