N,N'-(2,5-dichloro-1,4-phenylene)bis[4-[[2-chloro-5-(trifluoromethyl)phenyl]azo]-3-hydroxynaphthalene-2-carboxamide]

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

Analogue substance: Pigment Red 220 (not specified form):

 

A GLP-compliant investigation of the toxicological effects resulting from repeated oral-gavage administration to rats was performed following OECD guideline 422 without deviations (BASF 2012b). Pigment Red 220 (CAS 68259-05-2)was administered in water as vehicle at dosages of 100, 300, and 1000 mg/kg body weight/day, and controls received the vehicle only. Pigment Red 220 was administered to male rats for 29 days and to female rats for 14 days prior to pairing, through the pairing and gestation periods until the F1 generation reached day 4 post partum. Treatment with the test item up and including 1000 mg/kg bw/day did not reveal any clinical signs or histological findings and did not affect reproduction and development.

All dose-treated males and females had dose-related reddish discolored feces during the treatment period. This finding is considered to be a typical effect resulting from oral administration of a red dyestuff and not adverse.

Based on these results a general NOAEL (No Observed Adverse Effect Level) was considered to be 1000 mg/kg body weight/day.

The NOEL (No Observed Effect Level) for reproduction/developmental toxicity was considered to be 1000 mg/kg body weight/day.

Link to relevant study records

Reference

Reference 1

Endpoint:

screening for reproductive / developmental toxicity

Data waiving:

other justification

Justification for data waiving:

other:

Reference 2

Endpoint:

screening for reproductive / developmental toxicity

Remarks:

based on test type (migrated information)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2011-2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP and OECD 422 compliant study with well-characterized test material.

Qualifier:

according to guideline

Guideline:

OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Species:

rat

Strain:

other: RccHanTM: WIST(SPF)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Laboratories, B.V., 5961 NM Horst / Netherlands
- Age at study initiation: 11 weeks
- Weight at study initiation: Males: 315 to 364 g, Females: 182 to 212 g
- Fasting period before study: none
- Housing: individually
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 30 - 70%
- Air changes (per hr): 10 - 15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 2011-12-15 To: 2012-02-06

Route of administration:

oral: gavage

Vehicle:

water

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: The dose formulations were prepared fresh daily using the test item as supplied by the Sponsor.


VEHICLE
- Concentration in vehicle: as adjusted to dose
- Amount of vehicle (if gavage): 10 mL/kg body weight

Details on mating procedure:

- M/F ratio per cage: (1:1)
- Length of cohabitation: 14 days
- Proof of pregnancy: vaginal plug or sperm in vaginal smear referred to as day 0 of pregnancy
- After 14 days of unsuccessful pairing no further mating was performed.
- After successful mating each pregnant female was caged (how): groups of three or four

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

On the first treatment day samples from the control group as well as three samples (top, middle
and bottom) of about 2 g of each concentration were taken prior to dosing for analysis of
concentration and homogeneity. During the last week of the treatment, samples were taken from
the middle to confirm concentration.

Duration of treatment / exposure:

Males: Minimum 4 weeks
Females: Approximately 6 weeks

Frequency of treatment:

daily

Dose / conc.:

1 000 mg/kg bw/day (actual dose received)

Dose / conc.:

300 mg/kg bw/day (actual dose received)

Dose / conc.:

100 mg/kg bw/day (actual dose received)

Dose / conc.:

0 mg/kg bw/day (actual dose received)

No. of animals per sex per dose:

10

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Based on information on a substance of similar structure.

Positive control:

Not required.

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily (twic for mortality)

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Once prior to the first administration of the test item and weekly thereafter (in the gestation period on day 0, 6, 13 and 20 post coitum), detailed clinical observations were performed outside the home cage in a standard arena.
- Cage side observations : changes in skin, fur, eyes, mucous membranes, occurrence of secretions and excretions, and autonomic activity (e.g.
lacrimation, piloerection, pupil size, and unusual respiratory pattern). Changes in gait, posture and response to handling as well as the presence of clonic or tonic movements, stereotypies or bizarre behavior were also reported.


BODY WEIGHT: Yes
- Time schedule for examinations: daily

Oestrous cyclicity (parental animals):

not examined

Sperm parameters (parental animals):

During histopathology, special emphasis was made on the stages of spermatogenesis and histopathology of interstitial cell structure.
Testis weight and epididymis weight were determined.

Litter observations:

STANDARDISATION OF LITTERS
not required

PARAMETERS EXAMINED
The following parameters were examined: number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, weight gain, physical or behavioural abnormalities

GROSS EXAMINATION OF DEAD PUPS: yes, for external and internal abnormalities; possible cause of death was not determined for pups born or found dead.

Postmortem examinations (parental animals):

SACRIFICE
- Male animals: All surviving animals [describe when, e.g. as soon as possible after the last litters in each generation were produced.]
- Maternal animals: All surviving animals [describe when, e.g. after the last litter of each generation was weaned.]

GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations including the cervical, thoracic, and abdominal viscera.

HISTOPATHOLOGY / ORGAN WEIGHTS
As required for a full subacute oral toxicity study.

The number of implantation sites and corpora lutea was recorded for all dams with litters. The uteri of apparently non-pregnant females were placed in a solution of ammonium sulfide to visualize possible hemorrhagic areas of implantation sites

Postmortem examinations (offspring):

SACRIFICE
- The F1 offspring was sacrificed at 4 days of age.

GROSS NECROPSY
- Gross necropsy consisted of external examinations.

HISTOPATHOLOGY / ORGAN WEIGTHS
not examined.

Statistics:

Means and standard deviations of various data were calculated.
• The Dunnett-test (many to one t-test) based on a pooled variance estimate was applied if the variables could be assumed to follow a normal distribution for the comparison of the treated groups and the control groups for each sex.
• The Steel-test (many-one rank test) was applied instead of the Dunnett-test when the data could not be assumed to follow a normal distribution.
• Fisher's exact-test was applied if the variables could be dichotomized without loss of information.

Reproductive indices:

Percentage mating = ( Females mated / Females paired) * 100
Fertility index = ( Females achieving a pregnancy / Females paired) * 100
Conception rate = ( Females achieving a pregnancy / Females mated) * 100
Gestation index = ( Number of females with living pups / Females pregnant) * 100
Birth index = (pups born alive / number of implantations) * 100
post-implantation loss

Offspring viability indices:

Viability index (%) = number of live pups on day 4 after birth/number of live pups on the day of birth x 100

Clinical signs:

no effects observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Reproductive function: oestrous cycle:

not examined

Reproductive function: sperm measures:

no effects observed

Reproductive performance:

no effects observed

All animals mated within the first pairing period. The median and mean precoital times were unaffected by treatment with the test item. Mean
precoital times were 4.3, 2.5, 2.8 and 2.8 days in groups 1, 2, 3 and 4, respectively. The median precoital time was 3 days in all groups.
Two females in the control group and one female in groups 2, 3 and 4, respectively, were not pregnant. This finding was considered to be incidental. As a result the fertility index in the control group was 81.8% and 90.9% in groups 2 - 4.

Mean number of corpora lutea per dam (determined at necropsy) was similar in all groups (15.8, 14.4, 15.3 and 15.6 in order of ascending dose level) and gave no indication of a test item-related effect.

The mean duration of gestation was unaffected by exposure to the test item. Mean duration of gestation was 21.3, 21.6, 21.4 and 21.4 days, in order of ascending dose level.

No effects on implantation rate or post-implantation loss were observed at any dose level.
The mean number of implantations per dam was similar in all groups (13.8, 12.2, 13.9 and 13.1 in order of ascending dose levels). The mean incidence of post-implantation loss as a percentage of total implantations was 4.0, 8.2, 5.4 and 7.6%, in order of ascending dose level.

Dose descriptor:

NOAEL

Effect level:

>= 1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

clinical signs

body weight and weight gain

organ weights and organ / body weight ratios

histopathology: non-neoplastic

reproductive function (sperm measures)

Clinical signs:

no effects observed

Mortality / viability:

no mortality observed

Body weight and weight changes:

no effects observed

Sexual maturation:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

no effects observed

Histopathological findings:

not examined

No effects on litter size were observed at any dose level.
Mean litter size at first litter check was 13.2, 11.6, 13.1 and 12.1 pups in order of ascending dose levels. No dead pups at first litter check were recorded in any group.
No effects on postnatal loss were observed at any dose level.
Incidentally, 2, 2 and 1 pups were lost in groups 1, 2 and 4, respectively, between day 0 - 4 post partum.

No test item-related abnormal findings were noted at first litter check or during the first 4 days post partum.
Mean pup weights on day 0 and day 1 post partum were unaffected by treatment with the test
item. On day 1 post partum mean pup weights were 5.7, 6.0, 5.8 and 6.0 g in order of ascending
dose level. Also mean body weight gain was similar in all groups and on day 4 post partum no
effects on mean body weights were recorded.
No findings were noted at macroscopic examination of F1 pups.

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

>= 1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

viability

mortality

body weight and weight gain

Reproductive effects observed:

not specified

Conclusions:

No indication of toxicity to reproduction in rats were observed in the screening study at doses of up to 1000 mg/kg bw .

Reference 3

Endpoint:

screening for reproductive / developmental toxicity

Remarks:

based on test type (migrated information)

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH
[Please provide information for all of the points below. Indicate if further information is included as attachment to the same record, or elsewhere in the dataset (insert links in 'Cross-reference' table)]

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
Please refer to attached read across justification document (Chapter 13).

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
Please refer to attached read across document (Chapter 13).

3. ANALOGUE APPROACH JUSTIFICATION
Please refer to attached read across justification document (Chapter 13).

4. DATA MATRIX
Please refer to attached read across justification document (Chapter 13).

Reason / purpose for cross-reference:

read-across source

Clinical signs:

no effects observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Reproductive function: oestrous cycle:

not examined

Reproductive function: sperm measures:

no effects observed

Reproductive performance:

no effects observed

All animals mated within the first pairing period. The median and mean precoital times were unaffected by treatment with the test item. Mean
precoital times were 4.3, 2.5, 2.8 and 2.8 days in groups 1, 2, 3 and 4, respectively. The median precoital time was 3 days in all groups.
Two females in the control group and one female in groups 2, 3 and 4, respectively, were not pregnant. This finding was considered to be incidental. As a result the fertility index in the control group was 81.8% and 90.9% in groups 2 - 4.

Mean number of corpora lutea per dam (determined at necropsy) was similar in all groups (15.8, 14.4, 15.3 and 15.6 in order of ascending dose level) and gave no indication of a test item-related effect.

The mean duration of gestation was unaffected by exposure to the test item. Mean duration of gestation was 21.3, 21.6, 21.4 and 21.4 days, in order of ascending dose level.

No effects on implantation rate or post-implantation loss were observed at any dose level.
The mean number of implantations per dam was similar in all groups (13.8, 12.2, 13.9 and 13.1 in order of ascending dose levels). The mean incidence of post-implantation loss as a percentage of total implantations was 4.0, 8.2, 5.4 and 7.6%, in order of ascending dose level.

Dose descriptor:

NOAEL

Effect level:

>= 1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

clinical signs

body weight and weight gain

organ weights and organ / body weight ratios

histopathology: non-neoplastic

reproductive function (sperm measures)

Clinical signs:

no effects observed

Mortality / viability:

no mortality observed

Body weight and weight changes:

no effects observed

Sexual maturation:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

no effects observed

Histopathological findings:

not examined

No effects on litter size were observed at any dose level.
Mean litter size at first litter check was 13.2, 11.6, 13.1 and 12.1 pups in order of ascending dose levels. No dead pups at first litter check were recorded in any group.
No effects on postnatal loss were observed at any dose level.
Incidentally, 2, 2 and 1 pups were lost in groups 1, 2 and 4, respectively, between day 0 - 4 post partum.

No test item-related abnormal findings were noted at first litter check or during the first 4 days post partum.
Mean pup weights on day 0 and day 1 post partum were unaffected by treatment with the test
item. On day 1 post partum mean pup weights were 5.7, 6.0, 5.8 and 6.0 g in order of ascending
dose level. Also mean body weight gain was similar in all groups and on day 4 post partum no
effects on mean body weights were recorded.
No findings were noted at macroscopic examination of F1 pups.

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

>= 1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

viability

mortality

body weight and weight gain

Reproductive effects observed:

not specified

Conclusions:

No indication of toxicity to reproduction in rats were observed in the screening study at doses of up to 1000 mg/kg bw .

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Study duration:

subacute

Species:

rat

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Effects on developmental toxicity

Description of key information

Disazocondensation red pigments are too large and insoluble for significant systemic uptake. Therefore, no hazard on developmental toxicity is possible. No adverse effects were observed in the screening study at the limit dose (OECD 422) for Pigment Red 220 (CAS 68259-05-2, BASF 2012b).

Link to relevant study records

Reference

Endpoint:

developmental toxicity

Data waiving:

other justification

Justification for data waiving:

other:

Species:

rat

Abnormalities:

not specified

Developmental effects observed:

not specified

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Study duration:

subacute

Species:

rat

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no study available

Additional information

Pigment Red 220 (CAS 68259-05-2, 926 g/mol)

 A GLP-compliant investigation of the toxicological effects resulting from repeated oral-gavage administration to rats was performed following OECD guideline 422 without deviations (Harlan 2012). Pigment Red 220 (CAS 68259-05-2)was administered in water as vehicle at dosages of 100, 300, and 1000 mg/kg body weight/day, and controls received the vehicle only. Pigment Red 220 was administered to male rats for 29 days and to female rats for 14 days prior to pairing, through the pairing and gestation periods until the F1 generation reached day 4 post partum. Treatment with the test item up and including 1000 mg/kg bw/day did not reveal any clinical signs or histological findings and did not affect reproduction and development.

All dose-treated males and females had dose-related reddish discolored feces during the treatment period. This finding is considered to be a typical effect resulting from oral administration of a red dyestuff and not adverse.

Based on these results a general NOAEL (No Observed Adverse Effect Level) was considered to be 1000 mg/kg body weight/day.

The NOEL (No Observed Effect Level) for reproduction/developmental toxicity was considered to be 1000 mg/kg body weight/day.

 

 

Justification for classification or non-classification

Classification, Labelling, and Packaging Regulation (EC) No. 1272/2008

The available screening study in combination with toxicokinetic considerations is reliable and suitable for classification purposes under Regulation 1272/2008. As a result the substance is not considered to be classified for fertility or developmental toxicity under Regulation (EC) No. 1272/2008, as amended for the third time in Directive EC 618/2012.