Mixture of reaction products of substituted copper phthalocyanine, hydrogenated resin acids, alkaline earth metal chlorides, and copper phthalocyanine

Developmental toxicity / teratogenicity

Administrative data

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

April-June 2009

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Performed according to OECD 421 and GLP principles.

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Cross-reference 2

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Report date:

2009

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

other: OECD 421

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: US EPA OPPTS 870.3550

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

B508

IUPAC Name:

B508

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material : B508
- Physical state: powder

Test animals

Species:

rat

Strain:

other: Crl:WI(Han)

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: 11 wks
- Weight at study initiation: 295-320 g for males; 182-208 g for females
- Housing: in Macrolon cages with sterilized sawdust as bedding material and paper as cage-enrichment; 5 animals/sex/cage pre-mating; 1 male and 1 female per cage during mating; at post-mating males were kept with a maximum of 5 animals/cage and females were individually housed.
- Diet: free access to pelleted rodent diet
- Water: free access to tap-water
- Acclimation period: at least 5 days prior to start of treatment

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.2-21.4
- Humidity (%): 33-76
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 29 April - 17 June 2009

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: 0.5 w/v% methylcellulose

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: prepared daily within 6 hours prior to dosing and homogenized to a visually acceptable level

VEHICLE
- Justification for use and choice of vehicle (if other than water): based on information provided by the sponsor and trial formulations at the test lab.
- Amount of vehicle (if gavage): 5 ml/kg bw

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The concentrations analysed at 50 and1000 mg/kg bw/d, based on copper and strontium analysis using ICP-MS, were in agreement with the target concentrations (i.e. mean accuracies between 85 and 115%). The concentrations analysed at 250 mg/kg bw/d, based on copper analysis, were slightly above target concentration (mean accuracy 116%) but the result was accepted since the corresponding result based on strontium analysis was well within the criterion range.

Details on mating procedure:

- M/F ratio per cage: 1/1 (one female was cohabitated with one male of the same treatment group, avoiding sibling mating)
- Length of cohabitation: maximum 14 days with one male
- Proof of pregnancy: vaginal plug or sperm in vaginal smear referred to as day 0 of pregnancy
- Further matings after an unsuccessful attempt: no

Duration of treatment / exposure:

Males were exposed for 28 days, i.e. 2 weeks prior to mating, during mating, and up to the day prior to necropsy. Females were exposed for 40 to 49 days, i.e. during 2 weeks prior to mating, during mating, during post-coitum, and during at least 3 days of lactation.

Frequency of treatment:

Once daily for 7 days per week. Animals were dosed up to the day prior to scheduled necropsy.

Duration of test:

Pups were killed on day 4 of lactation or shortly thereafter.

Doses / concentrations

Remarks:

Doses / Concentrations:
50, 250 and 1000 mg/kg bw/d
Basis:
actual ingested

No. of animals per sex per dose:

9 litters/dose

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: the selected dose levels are based on the outcome of the 28-day repeated dose study (CERI B11-0899; see 7.5.1). The NOAEL of this study was 1000 mg/kg bw/d based on the absence of effects.

Examinations

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: at least twice daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: at least once daily

BODY WEIGHT: Yes
- Time schedule for examinations: Males and females were weighed on the first day of exposure and weekly thereafter. Mated females were weighed on Days 0, 4, 7, 11, 14, 17 and 20 post-coitum, and during lactation on Days 1 and 4.

FOOD CONSUMPTION: Yes
Weekly, for males and females. Food consumption was not recorded during the breeding period. Food consumption of mated females was measured on day 0, 4, 7, 11, 14, 17 and 20 post-coitum and during lactation on day 1 and 4.

WATER CONSUMPTION : No
Subjective appraisal was maintained during the study, but no quantitative investigation was introduced as no effect was suspected.

Ovaries and uterine content:

not examined

Fetal examinations:

yes, foetal weight

Statistics:

The following statistical methods were used to analyze the data:
- If the variables could be assumed to follow a normal distribution, the Dunnett-test (Dunnett, 1955) (many-to-one t-test) based on a pooled variance estimate was applied for the comparison of the treated groups and the control groups for each sex.
- The Steel-test (Miller, 1981) (many-to-one rank test) was applied if the data could not be assumed to follow a normal distribution.
- The Fisher Exact-test (Fisher 1950) was applied to frequency data.
- The implantation index, number of live pups born, sex ratio and delivery index were subjected to the Kruskal-Wallis nonparametric ANOVA test (Kruskal, 1952) to determine intergroup difference. If the results of the ANOVA were significant (p<0.05), the Wilcoxon test (Wilcoxon, 1945) was applied to the data to compare the treated groups to the control group.

All tests were two-sided and in all cases p < 0.05 was accepted as the lowest level of significance.
Group means were calculated for continuous data and medians were calculated for discrete data (scores) in the summary tables. Test statistics were calculated on the basis of exact values for means and pooled variances. Individual values, means and standard deviations may be rounded off before printing. Therefore, two groups may display the same printed means for a given parameter, yet display different test statistics values.

No statistical analysis was performed on histopathology findings.

Results and discussion

Results: maternal animals

Maternal developmental toxicity

Details on maternal toxic effects:

Maternal toxic effects:no effects

Effect levels (maternal animals)

Dose descriptor:

NOAEL

Effect level:

>= 1 000 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

other: developmental toxicity

Results (fetuses)

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Fetal abnormalities

Abnormalities:

not specified

Overall developmental toxicity

Developmental effects observed:

not specified

Any other information on results incl. tables

There was no treatment-related change in postnatal loss, viability index, clinical symptoms, sex ratio, pup weight or external appearance at necropsy.

Applicant's summary and conclusion

Conclusions:

In a screening reproduction/developmental toxicity study in rats the NOAEL for developmental toxicity was established as being at least 1000 mg/kg bw/d.

Executive summary:

Rats were dosed with 0, 50, 250 or 1000 mg/kg bw/d B508 via gavage in a screening reproduction/developmental toxicity study performed according to OECD 421 and GLP principles.

There was no treatment-related change in postnatal loss, viability index, clinical symptoms, sex ratio, pup weight or external appearance at necropsy. Therefore, the NOAEL for developmental toxicity was established to be at least 1000 mg/kg bw/d.