Pyrogallol

Developmental toxicity / teratogenicity

Administrative data

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Data source

Reference

Reference Type:

publication

Title:

TERATOLOGY AND PERCUTANEOUS TOXICITY STUDIES ON HAIR DYES

Author:

Burnett C, Goldenthal EI, Harris SB, Wazeter FX, Strausburg J, Kapp R, Voelker R.

Year:

1976

Bibliographic source:

J Toxicol Environ Health. 1976. 1(6): 1027-1040.

Materials and methods

Test guideline

Qualifier:

no guideline available

Principles of method if other than guideline:

- Principle of test: The chronic toxicological potential following skin painting in mice was evaluated for oxidative and nonoxidative hair dyes formulation together with a control groups which were shaved only and received no applications
- Short description of test conditions: dermal exposure was assessed; groups of male and female Swiss mice were treated one time weekly for at least 20 mo with one dose level of each dye. The oxidative dyes were mixed 1:1 with 6% hydrogen peroxide before treatment and the three semipermanent formulations were applied without dilution
- Parameters analysed / observed: Toxicological and carcinogenic effects were analysed

GLP compliance:

not specified

Test material

Test material information

Constituent 1

Reference substance name:

Pyrogallol

EC Number:

201-762-9

EC Name:

Pyrogallol

Cas Number:

87-66-1

Molecular formula:

C6H6O3

IUPAC Name:

benzene-1,2,3-triol

impurity 1

Reference substance name:

unknown impurities

IUPAC Name:

unknown impurities

Test material form:

not specified

Details on test material:

not specified

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material:
The Clairol Research Laboratories (Stamford, Conn.) provided seven oxidative hair dye formulations, which were identified as 7401, 7402, 7403, 7404, 7405, 7406, and P-21. They also provided three semipermanent
hair dye formulations, coded P-22, P-23, and P-24. The remaining two oxidative hair dye formulations, P-25 and P-26, were provided by L'Oreal Inc. (Clark, N.J.).
- Expiration date of the lot/batch:
not specified
- Purity test date:
not specified

RADIOLABELLING INFORMATION (if applicable)
- Radiochemical purity:
not applicable
- Specific activity:
- Locations of the label:
- Expiration date of radiochemical substance:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material:
not specified
- Stability under test conditions:
not specified
- Solubility and stability of the test substance in the solvent/vehicle:
not specified
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium:

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing:
not specified
- Preliminary purification step (if any):
not specified
- Final dilution of a dissolved solid, stock liquid or gel:
not specified
- Final preparation of a solid:
not specified

FORM AS APPLIED IN THE TEST (if different from that of starting material): solutions

TYPE OF BIOCIDE/PESTICIDE FORMULATION (if applicable): not applicable

Test animals

Species:

rat

Strain:

other: Charles River CD

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: not specified
- Age at study initiation: not specified
- Weight at study initiation: not specified
- Fasting period before study: not specified
- Housing: The mated females were housed individually in temperature- and humidity-controlled rooms during the study and were weighed on the days the dyes were administered.
- Diet (e.g. ad libitum): Ralston Purina Laboratory Chow available ad libitum.
- Water (e.g. ad libitum): available ad libitum.
- Acclimation period: not specified

ENVIRONMENTAL CONDITIONS
- Temperature (°C): not specified
- Humidity (%): not specified
- Air changes (per hr): not specified
- Photoperiod (hrs dark / hrs light): not specified

IN-LIFE DATES: From: To: not specified

Administration / exposure

Route of administration:

dermal

Remarks on MMAD:

not relevant: the substance was applied topically

Details on exposure:

TEST SITE
- Area of exposure: The hair at the site of application on the dorso-scapular area was shaved closely the day before the solutions were applied
- % coverage: not specified
- Type of wrap if used: not specified
- Time intervals for shavings or clipplings: not specified

REMOVAL OF TEST SUBSTANCE
- Washing (if done):
- Time after start of exposure:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): not specified
- Concentration (if solution): not specified
- Constant volume or concentration used: yes/no not specified
- For solids, paste formed: yes/no not specified

VEHICLE
- Justification for use and choice of vehicle (if other than water):
- Amount(s) applied (volume or weight with unit):
- Concentration (if solution):
- Lot/batch no. (if required):
- Purity:

USE OF RESTRAINERS FOR PREVENTING INGESTION: yes/no

Analytical verification of doses or concentrations:

not specified

Details on mating procedure:

not specified

Duration of treatment / exposure:

not specified

Frequency of treatment:

not specified

Duration of test:

not specified

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

2 other: ml/kg

Remarks:

dose of formulation applied (pyrogallol was a constituent of the formulation named P-21: 0.4%)

Doses / concentrations 2

Dose / conc.:

8 mg/kg bw/day

Remarks:

calculated pyrogallol dose

No. of animals per sex per dose:

20 (only female)

Control animals:

yes

Details on study design:

- Dose selection rationale: previous pitol study
- Rationale for animal assignment (if not random): random
- Other: -

Examinations

Maternal examinations:

not relevant

Ovaries and uterine content:

The uteri were examined, corpora lutea of pregnancy counted, and the number, distribution, and location of live, dead, and resorbed fetuses
recorded.

Fetal examinations:

All fetuses were examined for gross anomalies, sexed, and weighed. Approximately one-third the fetuses from each litter were fixed in Bouin's solution and subsequently examined for visceral anomalies by razor blade sectioning (Wilson and Warkany, 1965). The remaining fetuses
in each litter were fixed in 95% ethyl alcohol, eviscerated, cleared, stained with KOH-alizarin red S (Staples and Schnell, 1964), and examined for skeletal anomalies. Fetal examinations were performed in a random order with treatment group identity unknown to the examiner

Statistics:

All statistical analyses compared the treatment groups with the control groups. The number of females exhibiting resorption sites, number of females exhibiting two or more resorptions, number of dead or resorbed fetuses, and the number of fetuses with soft-tissue or skeletal anomalies and accessory ribs was compared using chi-square test criterion with Yates' correction on 2 X 2 contingency tables as described by Steel and Torrie (1960) or Fisher's exact probability test (Siegel, 1956) as appropriate to judge the significance of difference.
The mean number of corpora lutea, implantation sites, live fetuses, and resorption sites was compared by analysis of variance (one-way classification)
as described by Steel and Torrie (1960) using Dunnett's (1964) multiple comparison tables to judge the significance of differences.
The live fetal weights were compared by analysis of variance (hierarchal classification) as described by Steel and Torrie (1960) using Dunnett's (1964) multiple comparison tables to judge the significance of differences.
Statistically significant differences between groups were judged valid only when there were significant differences between any one of the dye treated groups and each of the three untreated control groups.

Indices:

not specified

Historical control data:

not specified

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:

not examined

Dermal irritation (if dermal study):

no effects observed

Mortality:

not examined

Body weight and weight changes:

not examined

Food consumption and compound intake (if feeding study):

not examined

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

not examined

Haematological findings:

not examined

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

not examined

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

not examined

Histopathological findings: neoplastic:

not examined

Other effects:

not examined

Maternal developmental toxicity

Number of abortions:

not examined

Pre- and post-implantation loss:

not examined

Total litter losses by resorption:

not examined

Early or late resorptions:

not examined

Dead fetuses:

not examined

Changes in pregnancy duration:

not examined

Changes in number of pregnant:

not examined

Other effects:

no effects observed

Description (incidence and severity):

Changes in female body weights were similar for rats in the untreated controls and all dye-treated groups
Mean food consumption for all groups throughout gestation was similar.

Details on maternal toxic effects:

Maternal toxicity was not observed

Results (fetuses)

Fetal body weight changes:

no effects observed

Reduction in number of live offspring:

no effects observed

Changes in sex ratio:

no effects observed

Changes in litter size and weights:

no effects observed

Changes in postnatal survival:

no effects observed

External malformations:

no effects observed

Skeletal malformations:

effects observed, non-treatment-related

Description (incidence and severity):

Effects observed for a formulation no-pyrogallol-containing.

Visceral malformations:

not examined

Other effects:

no effects observed

Description (incidence and severity):

fetal soft tissue anomalies: no significant differences.

Details on embryotoxic / teratogenic effects:

not reported

Effect levels (fetuses)

Dose descriptor:

NOAEC

Effect level:

ca. 8 mg/kg bw/day

Based on:

act. ingr.

Sex:

female

Basis for effect level:

changes in sex ratio

skeletal malformations

Remarks on result:

other: no embryotoxic or teratogenic effects observed at the dose tested

Fetal abnormalities

Abnormalities:

no effects observed

Description (incidence and severity):

no embryotoxic or teratogenic effects.

Overall developmental toxicity

Developmental effects observed:

no

Lowest effective dose / conc.:

8 mg/kg bw/day

Treatment related:

no

Applicant's summary and conclusion

Conclusions:

No significant differences in the number of corpora lutea, implantation sites, live fetuses, resorptions, or fetal soft tissue or skeletal anomalies were induced.
The teratogenicity of a hair dye formulation containing 0.4% Pyrogallol was evaluated using 20 Charles River CD female rats. The hair dye (2 ml/kg) was applied to the dorsoscapular area (shaved skin) of each animal on days 1,4, 7, 10, 13, 16, and 19 of gestation. Three groups of untreated rats (unshaved) served as controls. Animals in the positive control group were given acetylsalicylic acid (250 mg/kg) via gavage on days 6 to 16 of gestation. The dams were killed on day 20 of gestation via chloroform anesthesia, and fetuses were removed via cesarean section. One third of the fetuses from each litter were examined for visceral anomalies. The remaining fetuses were examined for skeletal anomalies. Toxic effects were not observed in experimental or control dams throughout the study. The mean numbers of corpora lutea, implantation sites, and live fetuses in experimental groups were not significantly different from those in control groups. There were also no significant differences in the nurnber of females with resorption sites and the mean number of resorptions per pregnancy. The incidence of fetal soft tissue and skeletal anomalies in experimental groups was not significantly different from that of negative control groups. A significant increase in the number of fetuses with skeletal and soft tissue anomalies and in the number of dead or resorbed fetuses was observed in the positive control group

Executive summary:

Twelve hairdye formulations were tested by topical application twice weekly for 13 wk for teratologic effects following applications to groups of 20 pregnant Charles River CD rats on days 1, 4, 7, 10, 13, 16, and 19 of gestation. The three semipermanent formulations were applied as is, and the nine oxidationdyes were mixed 1:1 with 6% hydrogen peroxide just prior to application, as in normal use. The formulations induced a broad spectrum of dye sand dye intermediates used or considered useful in oxidative and semipermanent haircolor products. In theteratology study no biologically significant soft tissue or skeletal changes were noted. Similarly, the mean numbers of corpora lutea, implantation sites, live fetuses, and resorptions per pregnancy, as well as numbers of litters with resporptions, were not significantly affected by the dye treatment.