Registration Dossier

Administrative data

Description of key information

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Data is from National Cancer Institute Technical report
Qualifier:
according to
Guideline:
other: as below
Principles of method if other than guideline:
Subchronic toxicity tests were conducted with rats
GLP compliance:
not specified
Species:
rat
Strain:
Fischer 344
Sex:
male/female
Details on test animals and environmental conditions:
- Source: A. R. Schmidt, Madison, Wisconsin, and Laboratory Supply Company, Inc., Indianapolis, Indiana.- Age at study initiation: 4 weeks old- Weight at study initiation: No data- Fasting period before study: Housing: polycarbonate cages suspended from aluminum racks. Ab-sorb-dri® hardwood chip bedding- Diet (e.g. ad libitum): Wayne Lab-Blox meal, ad libitum - Water (e.g. ad libitum): Acidulated water (pH 2.5) was supplied to animals in water bottles filled by an automated metering device, ad libitum- Acclimation period: quarantined for 2 weeks prior to initiationENVIRONMENTAL CONDITIONS- Temperature (°C): 22° to 26°C- Humidity (%): 45 and 55 percent- Air changes (per hr): Incoming air was filtered through HEPA filters, at a rate of 12 to 15 complete changes of room air per hour.- Photoperiod (hrs dark / hrs light): Fluorescent lighting was provided 8 hours per day (9:00 a.m. to 5:00 p.m.).IN-LIFE DATES: From: To:
Route of administration:
oral: feed
Vehicle:
other: basal laboratory diet
Details on oral exposure:
DIET PREPARATIONThe chemical was removed from its container and a proper amount was blended with an aliquot of the ground feed using a mortar and pestle. Once visual homogeneity was attained, the mixture was placed in a 6 kg capacity Patterson-Kelley standard model twin-shell stainless steel V-blender along with the remainder of the feed to be prepared. After 20 minutes of blending, the mixtures were placed in double plastic bags and stored in the dark at 4°C. The mixture was prepared once weekly.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Doses were analysed spectrophotometrically
Duration of treatment / exposure:
4 weeks followed by 2 weeks observation
Frequency of treatment:
Daily
Remarks:
Doses / Concentrations:31.5, 68.0, 146.5,315.5 and 680.0 mg/kgbwBasis:no data
No. of animals per sex per dose:
five males and five females in six dose groups
Control animals:
yes
Details on study design:
No data
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes / No / No data: No dataTime schedule: No data- Cage side observations checked in table [No.?] were included. No dataDETAILED CLINICAL OBSERVATIONS: Yes / No / No data: No data Time schedule: No dataBODY WEIGHT: Yes / No / No data: YesTime schedule for examinations: No dataFOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): No dataFood consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes / No / No data: No dataCompound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes / No / No data: No dataFOOD EFFICIENCY:Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: Yes / No / No data: No dataWATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes / No / No data: No dataTime schedule for examinations: No dataOPHTHALMOSCOPIC EXAMINATION: Yes / No / No data: No dataTime schedule for examinations: No dataDose groups that were examined:No dataHAEMATOLOGY: Yes / No / No data: No dataTime schedule for collection of blood: No dataAnaesthetic used for blood collection: Yes (identity) / No / No data: No dataAnimals fasted: Yes / No / No data: No dataHow many animals:- Parameters checked in table [No.?] were examined.: No dataCLINICAL CHEMISTRY: Yes / No / No dataTime schedule for collection of blood: No dataAnimals fasted: Yes / No / No data: No dataHow many animals: No dataParameters checked in table [No.?] were examined.: No dataURINALYSIS: Yes / No / No dataTime schedule for collection of urine: No dataMetabolism cages used for collection of urine: Yes / No / No data: No dataAnimals fasted: Yes / No / No data: No dataParameters checked in table [No.?] were examined.: No dataNEUROBEHAVIOURAL EXAMINATION: Yes / No / No data: No data
Sacrifice and pathology:
No data
Other examinations:
No data
Statistics:
No data
Clinical signs:
no effects observed
Description (incidence and severity):
No abnormal clinical signs were recorded for any rat group.
Mortality:
no mortality observed
Description (incidence):
No abnormal clinical signs were recorded for any rat group.
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:
not specified
Histopathological findings: non-neoplastic:
not specified
Histopathological findings: neoplastic:
not specified
Dose descriptor:
NOAEL
Effect level:
680 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No effects observed
Critical effects observed:
not specified
Conclusions:
Sub-chronic toxicity tests were conducted with both rats and mice. Rats were distributed among six groups, each consisting of five males and five females. 2-Nitro-p-phenylenediamine was incorporated into the basal laboratory diet and supplied ad libitum to five of the six rat groups in concentrations of 31.5, 68.0, 146.5,315.5 and 680.0 mg/kgbw. The remaining rat group served as a control group, receiving only the basal laboratory diet. No abnormal clinical signs were recorded for any rat group.The NOAEL for 2 nitro p phenylenediamine is 680.0mg/kgbw
Executive summary:

Subchronic toxicity tests were conducted with both rats and mice. Rats were distributed among six groups, each consisting of five males and five females. 2-Nitro-p-phenylenediamine was incorporated into the basal laboratory diet and supplied ad libitum to five of the six rat groups in concentrations of 31.5, 68.0, 146.5,315.5 and 680.0 mg/kg bw. The remaining rat group served as a control group, receiving only the basal laboratory diet.

The dosed dietary preparations were administered for a period of 4 weeks, followed by a 2-week observation period during which all animals were fed the basal laboratory diet. Individual body weights and food consumption data were recorded twice weekly throughout the study. Upon termination of the study all survivors were sacrificed and necropsied.

 

No abnormal clinical signs were recorded for any rat group. The NOAEL for 2 nitro p phenylenediamine is 680 mg/ kg bw.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed

Repeated dose toxicity: dermal - systemic effects

Link to relevant study records
Reference
Endpoint:
chronic toxicity: dermal
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Qualifier:
according to
Guideline:
other: as mentioned below
Principles of method if other than guideline:
To determine the repeated dose toxicity of the test chemical when applied dermally to rabbits
GLP compliance:
no
Species:
rabbit
Strain:
New Zealand White
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALSSource: No dataAge at study initiation: No dataWeight at study initiation: No data Fasting period before study: No dataHousing: metal cagesDiet (e.g. ad libitum):No dataWater (e.g. ad libitum):No dataAcclimation period: No dataENVIRONMENTAL CONDITIONSTemperature (°C): No dataHumidity (%): No dataAir changes (per hr):No dataPhotoperiod (hrs dark / hrs light):No dataIN-LIFE DATES: From: To:
Type of coverage:
other: on the dorsolateral aspects of the thoracic-lumbar area (one on each side of the midline)
Vehicle:
other: Yes
Details on exposure:
REMOVAL OF TEST SUBSTANCEWashing (if done):YesTime after start of exposure: 1 hourTEST MATERIALAmount(s) applied (volume or weight with unit): A 1 ml/kg dose of 1 :1 dye: hydrogen peroxidewas applied to alternating siteConcentration (if solution): 1 ml/kg Constant volume or concentration used: yes/no: Yes- For solids, paste formed: yes/no: No dataUSE OF RESTRAINERS FOR PREVENTING INGESTION: yes/no: Yes
Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
No data
Duration of treatment / exposure:
Applied topically two times a week for 13 weeks
Frequency of treatment:
Twice a week
Remarks:
Doses / Concentrations:1ml/kgBasis:
No. of animals per sex per dose:
6 rabbits/sex
Control animals:
other: Yes , three independent control groups of 12 rabbits
Details on study design:
No data
Positive control:
No data
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes / No / No data: No dataDETAILED CLINICAL OBSERVATIONS: Yes / No / No data: No dataDERMAL IRRITATION (if dermal study): Yes / No / No data: YesBODY WEIGHT: Yes / No / No data: YesTime schedule for examinations:WeeklyFOOD CONSUMPTION: No dataFOOD EFFICIENCY: No dataWATER CONSUMPTION: Yes / No / No data: No dataOPHTHALMOSCOPIC EXAMINATION: Yes / No / No data : No dataHAEMATOLOGY: Yes / No / No data: YesTime schedule for collection of blood: 0, 3, 7, and 13 wk.Anaesthetic used for blood collection: Yes (identity) / No / No data: No dataAnimals fasted: Yes / No / No data: No dataHow many animals: No dataParameters checked in table [No.?] were examined : blood count, methemoglobin, fasting blood sugar,CLINICAL CHEMISTRY: Yes / No / No data: YesTime schedule for collection of blood: 0, 3, 7, and 13 wk.Animals fasted: Yes / No / No data: No dataHow many animals: No data- Parameters checked in table [No.?] were examined: blood urea nitrogen, alkaline phosphatase, and serum glutamic oxaloacetic transaminaseURINALYSIS: Yes / No / No dataYesTime schedule for collection of urine: 0, 3, 7, and 13 wk. Metabolism cages used for collection of urine: Yes / No / No data: No data Animals fasted: Yes / No / No data: No dataParameters checked in table [No.?] were examined: Urinewas examined for color, pH, albumin, glucose, occult blood, and microscopic elements.NEUROBEHAVIOURAL EXAMINATION: Yes / No / No data: No data
Sacrifice and pathology:
GROSS PATHOLOGY: Yes (see table) / No / No data: No dataHISTOPATHOLOGY: Yes (see table) / No / No data: Yes
Other examinations:
No data
Statistics:
Statistical analysis of the data on body weight gains, hematology, clinical chemistries, and absolute and relative organ weights was performed using the analysis of variance F test and Student's t test.When variances differed significantly, Student's t test was modified (t1) and Cochran's approximation was utilized
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
No signs of toxicity were observed. Five control and five test animals died during the study due to complications resulting from cardiac puncture while collecting blood.
Dermal irritation:
not specified
Mortality:
mortality observed, treatment-related
Description (incidence):
No signs of toxicity were observed. Five control and five test animals died during the study due to complications resulting from cardiac puncture while collecting blood.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Body weight gains were equal to those of the controls.
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:
effects observed, treatment-related
Description (incidence and severity):
No significant differences were observed in hematological parameters
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
No significant differences were observed in clinical chemistry parameters
Urinalysis findings:
effects observed, treatment-related
Description (incidence and severity):
Urinalysis findings were “unremarkable,” and the urine was not discolored.
Behaviour (functional findings):
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Histopathological findings: neoplastic:
not specified
Details on results:
Gross pathology:No gross abnormalities were seen at necropsy, and no microscopic lesions were seen that were judged to be due to the administration of the hair dye formulations. The incidence and severity of disease processes common to laboratory rabbits was not affected by the experimental treatments.Histopathology:In no instance were any of the differences accompanied by histomorphologic evidence of toxicity. No gross abnormalities were seen at necropsy, and no microscopic lesions were seen that were judged to be due to the administration of the hair dye formulations. The incidence and severity of disease processes common to laboratory rabbits was not affected by the experimental treatments
Dose descriptor:
NOAEL
Effect level:
1 other: ml/kg
Based on:
test mat.
Sex:
male/female
Critical effects observed:
not specified
Conclusions:
A permanent hair dye formulation, containing 1.1 percent 2NPPD was mixed with an equal volume of 6 percent hydrogen peroxide was applied topically two times a week for 13 weeks to groups of 6 female and 6 male rabbits. A 1 ml/kg dose of 1:1 dye: hydrogen peroxide was applied to alternating sites. No signs of toxicity were observed.The NOAEL for repeated exposure to 2NPPD dermally in rabbits is 1ml/kg
Executive summary:

A permanent hair dye formulation, containing1.1percent 2NPPD was mixed with an equal volume of 6 percent hydrogen peroxide was applied topically two times a week for 13 weeks to groups of 6 female and 6 male rabbits. A 1 ml/kg dose of 1:1 dye: hydrogen peroxide was applied to alternating sites. The application sites of half of the animals were abraded once each week. The rabbits were restrained for an hour after application and were shampooed, rinsed, and dried.

 No signs of toxicity were observed. Body weight gains were equal to those of the controls. No significant differences were observed in hematological and clinical chemistry parameters.

The NOAEL for repeated exposure to 2NPPD dermally in rabbits is 1ml/kg
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed

Additional information

Various studies were reviewed to determine the toxic nature of the test compound 2 -nitro, p-phenylenediamine (CAS no 5307 -14 -2) upon repeated exposure orally/dermally. The summary is as mentioned below:

Repeated dose toxicity: Oral

Subchronic toxicity tests were conducted with both rats and mice (National Cancer Institute, 1979). Rats were distributed among six groups, each consisting of five males and five females. 2-Nitro-p-phenylenediamine (CAS no 5307-14-2) was incorporated into the basal laboratory diet and supplied ad libitum to five of the six rat groups in concentrations of 31.5, 68.0, 146.5, 315.5 and 680.0 mg/kg bw. The remaining rat group served as a control group, receiving only the basal laboratory diet. The dosed dietary preparations were administered for a period of 4 weeks, followed by a 2-week observation period during which all animals were fed the basal laboratory diet. Individual body weights and food consumption data were recorded twice weekly throughout the study. Upon termination of the study all survivors were sacrificed and necropsied. No abnormal clinical signs were recorded for any rat group. The NOAEL for 2 nitro p phenylenediamine is 680 mg/ kg bw.

In similar study on mice, the animals were distributed among ten groups, each consisting of five males and five females. 2-Nitro-p phenylene diamine (CAS no 5307-14-2) was incorporated into the basal laboratory diet and supplied ad libitum to eight of the ten mouse groups in concentrations of 162, 236 348, 510, 750, 1616, 2366 mg/kg bw. The two remaining mouse groups served as control groups, receiving only the basal laboratory diet. The dosed dietary preparations were administered for a period of 4 weeks, followed by a 2-week observation period during which all animals were fed the basal laboratory diet. Individual body weights and food consumption data were recorded twice weekly throughout the study. Upon termination of the study all survivors were sacrificed and necropsied. No abnormal clinical signs were recorded for any mouse group. The NOEL for 2 nitro p phenylenediamine is 2366 mg/kg bw.

Chronic toxicity tests were conducted with both rats and mice (National Cancer Institute, 1979). 2-Nitro-p-phenylenediamine (CAS no 5307-14-2) was incorporated into the basal laboratory diet and supplied ad libitum to dosed male rats. The highest dose concentration was estimated from a -sub chronic feeding study on groups of 5 male rats. Dose levels for chronic tests were decided as 55 mg/kg bw and 27.5 mg/kg bw. Dosed rat-s were supplied with feed containing 2-nitro-p-phenylenediamine for 78 weeks followed by a 27-week observation period. Animals were weighed immediately prior to initiation of the experiment and body weights were recorded once a week for the first 6 weeks, every 2 weeks for the next 12 weeks, and at monthly intervals thereafter. All animals were inspected twice daily for mortality. Food consumption data were collected at monthly intervals from 20% of the animals in each group. The animals were euthanized by carbon dioxide asphyxiation, and were immediately necropsied. The histopathologic examination consisted of gross and microscopic examination of all major tissues, organs, and gross lesions taken from sacrificed animals and, whenever possible, from animals found dead. No abnormal clinical signs were recorded. The NOAEL for 2 nitro p phenylenediamine in Fischer 344 male rats is 55 mg/kg bw.

In a similar chronic toxicity study, 2-Nitro-p-phenylenediamine (CAS no 5307-14-2) was incorporated into the basal laboratory diet and supplied ad libitum to dosed male rats. The highest dose concentration was estimated from a sub chronic feeding study on groups of 5 male rats. Dose levels for chronic tests were decided as 55 mg/kg bw and 110 mg/kg bw. Dosed rats were supplied with feed containing 2-nitro-p-phenylenediamine for 78 weeks followed by a 27-week observation period. Animals were weighed immediately prior to initiation of the experiment and body weights were recorded once a week for the first 6 weeks, every 2 weeks for the next 12 weeks, and at monthly intervals thereafter. All animals were inspected twice daily for mortality. Food consumption data were collected at monthly intervals from 20% of the animals in each group. The animals were euthanized by carbon dioxide asphyxiation, and were immediately necropsied. The histopathologic examination consisted of gross and microscopic examination of all major tissues, organs, and gross lesions taken from sacrificed animals and, whenever possible, from animals found dead. No abnormal clinical signs were recorded. The NOAEL for 2 nitro p phenylenediaminein Fischer 344 female rats is 110mg/kg bw.

Chronic toxicity test was conducted with mice (National Cancer Institute, 1979). 2-Nitro-p-phenylenediamine (CAS no 5307-14-2) was incorporated into the basal laboratory diet and supplied ad libitum to dosed mice. The highest dose concentration was estimated from a sub chronic feeding study on groups of 5 male and female mice. Dose levels for chronic tests were decided as 314.285 mg/kg bw and 628.571 mg/kg bw. Dosed mice were supplied with feed containing 2-nitro-p-phenylenediamine for 78 weeks followed by a 12-week observation period. Animals were weighed immediately prior to initiation of the experiment and body weights were recorded once a week for the first 6 weeks, every 2 weeks for the next 12 weeks, and at monthly intervals thereafter. All animals were inspected twice daily for mortality. Food consumption data were collected at monthly intervals from 20% of the animals in each group. The animals were euthanized by carbon dioxide asphyxiation, and were immediately necropsied. The histopathologic examination consisted of gross and microscopic examination of all major tissues, organs, and gross lesions taken from sacrificed animals and, whenever possible from animals found dead. No abnormal clinical signs were recorded. Distinct and consistent dose-related mean body weight depression was apparent in both male and female mice throughout the bioassay. A variety of non neoplastic lesions was present in both control and dosed animals. Hepatocellular adenoma and hepatocellular carcinoma occurred in a dose-related distribution in female mice. The NOAEL for 2 nitro p phenylenediamine in B6C3F1male mice is 628.571 mg/kg bw and 314.285 mg/kg bw in female mice.

Chronic toxicity tests were conducted with rats (Cosmetic Ingredients review, 1985). 10 male and 10 female rats were given a diet containing 500 mg/kg of 2 nitro p phenylenediamine (CAS no 5307-14-2) for 13 weeks. Changes in body weight, blood parameters, urine parameters and gross histopathology were observed. No changes were reported in all the parameters when compared with controls. The NOAEL can be reported as 500 mg/kg.

Composite of semi permanent hair coloring ingredients (0.24 percent 2NPPD) was administered in the diet to dogs for 2 yr to assess chronic toxicity (Wbrnick et al, 1975). Eighteen male and eighteen female purebred beagle dogs, 6-8 months of age, were acclimatized in air-conditioned quarters for 3 wk prior to initiation of dosing. They were housed individually and given food and water adlibitum. During this period, clinical parameters to be measured during the study were established twice for each dog and their eyes were examined once by a veterinary ophthamologist. The animals were divided into three groups of six males and six females each. The composite was incorporated into the diet at concentrations to give dosages of 0.0, 19.5, and 97.5 mg/kg/day. Diets were prepared fresh 7 days/wk and were usually consumed within 1 hr after feeding each day. Each animal was observed daily 7 days/wk for signs of toxic or pharmacologic effects. Individual records of body weight and food consumption were maintained on a weekly and daily basis, respectively. Physical examinations including funduscopic, EKG,blood pressure, pulse rate, and body temperature determinations were conducted initially and at 3, 6, 12, 18, and 24 mo. Hematological, blood chemical, and urinalysis parameters were determined on all dogs in the high dose and control groups and on three male and three female dogs in the low dose group at the same times No deaths. No adverse effects on weight gain or clinical, hematological, blood chemical, and urinalysis parameters. The NOAEL for 2 nitro p phenylene diamine in beagle dogs can be reported as 97.5mg/kg/day.Repeated dose toxicity: Dermal

A permanent hair dye formulation, containing1.1percent 2NPPD (CAS no 5307-14-2) was mixed with an equal volume of 6 percent hydrogen peroxide was applied topically two times a week for 13 weeks to groups of 6 female and 6 male rabbits (Burnett et al, 1976). A 1 ml/kg dose of 1:1 dye: hydrogen peroxide was applied to alternating sites. The application sites of half of the animals were abraded once each week. The rabbits were restrained for an hour after application and were shampooed, rinsed, and dried.No signs of toxicity were observed. Body weight gains were equal to those of the controls. No significant differences were observed in hematological and clinical chemistry parameters.The NOAEL for repeated exposure to 2NPPD dermally in rabbits is 1ml/kg.

 

A semi permanent hair colorant shampoo containing unspecified concentrations of 2NPPD (CAS no 5307-14-2) and 4NOPD was diluted 1:4:5 with water and acetone (Searle et al, 1977). A 0.4 ml dose of the mixture was usually applied twice a week to the clipped backs of groups of 26 male and female mice of strain A for 80 weeks. The statistical significances of the observed differences between control and treated groups were determined using the "logrank test". A survival time in weeks was determined for each mouse and in the context of the analysis, an "event" was recorded only if death was accompanied by the diagnosis of a tumor. Deaths from other causes were, however, accounted for in the figure for "extent of exposure The treatment was well tolerated by the A mice.The NOAEL for repeated exposure to 2NPPD dermally in mice is 0.4ml.

 

A hair dye composite containing 0.55 percent 2NPPD (CAS no 5307-14-2) was diluted 1:1 with a hydrogen peroxide activator and applied in doses of 1000, 2000, and 4000 mg/kg/day for 20 days to the shaved backs of groups of male and female rabbits (Cosmetic Ingredients Review, 1985). The application site was approximately 10 percent of the body surface.There was also a group of untreated control animals. The skin of 2 animals from each group was abraded prior to composite application. The rabbits were observed for a further 14 days after the test period. Two rabbits died during the test period in the 1000mg/kg group, 2 died in the 2000mg/kg group, 1 died in the 4000mg/kg dye-treated group, and 1 died in the control group. These deaths were attributed to naturally occurring disease; the incidence and severity of disease may have been increased due to the stress of the severe local skin reactions and the dosing procedure.No clinical signs of toxicity were observed. There were adverse effects on body weight in the treated rabbits during the test period, but body weights were comparable to the controls during the observation period. There were no significant adverse findings in the hematological and clinical chemistry parameters or in the urinalyses. No significant gross or microscopic alterations were observed in the tissues and organs of the rabbits killed at the end of the study or in any treated animals that died during the study, except for the skin.The NOAEL for repeated exposure to 2NPPD dermally in rabbits is 4000mg/kg day.

A permanent hair dye formulation containing 1.1percent 2NPPD (CAS no 5307-14-2) was mixed with an equal volume of 6 percent hydrogen peroxide (Cosmetic Ingredients Review, 1985). 2ml/kg of the hair dye was applied topically every third day of gestation for 19 days to a group of 20 pregnant rats.No signs of toxicity were observed. Maternal weight gain and feed consumption were similar to those of the controls. There was a change in the color of the hair and skin at the site of application, but no irritation was observed.The NOAEL for repeated exposure to 2NPPD dermally in pregnant rats is 2ml/kg.

 

Groups of 60 male and 60 female Swiss Webster mice, eight weeks of age, received topical applications of a semi-permanent hair dye formulation (7611) (WHO, 1993) 0.85% 2-nitro-para phenylenediamine (CAS no 5307-14-2).The formulation was applied at 0.05 ml/mouse three times a week for 20 months to a 1 cm2area of clipped shaved skin. Control animals were shaved only and received no treatment. All mice survived until termination of the experiment.The NOAEL for repeated exposure to 2NPPD dermally in mice is 0.05ml.

 

In a similar study in rats, groups of 60 male and female Sprague - Dawley rats, six to eight weeks of age, received topical applications of an oxidative hair dye formulation (7401) containing 1.1 % 1,4-diamino-2-nitrobenzene (2-nitro-p phenylenediamine, CAS no 5307-14-2). The formulation was diluted in an equal volume of 6% hydrogen peroxide before application, and 0.5 ml were applied to a shaved area of the back (approximately 2.5 cm in diameter) twice a week up until week 117.All rats survived until termination of the experiment.The NOAEL for repeated exposure to 2NPPD dermally in mice is 0.5ml

Justification for classification or non-classification