Registration Dossier

Toxicological information

Toxicity to reproduction

Currently viewing:

Administrative data

Endpoint:
one-generation reproductive toxicity
Remarks:
based on test type (migrated information)
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Conclusions drawn from a GLP experimental study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015
Report Date:
2015

Materials and methods

Principles of method if other than guideline:
Effects on male and female reproductive organs were studies in the 28 day repeated oral toxicity study
GLP compliance:
no
Remarks:
Deviation in test compared to the GLP guidelines for reproductive toxicity study
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
IUPAC name of the chemical: α,α-Bis[4-(dimethylamino)phenyl]-4 (phenylamino)naphthalene-1-methanol (C.I. Solvent Blue 4) [with < 0.1% of Michler's ketone
CAS Name: Solvent Blue 4 with less than 0.1% Michler's Ketone
CAS No: 6786-83-0
Nature of chemical: Organic

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
- Source: Central Animal Facility (CAF), NIPER, Sector-67, S.A.S. Nagar, 160 062, Punjab, India.
- Age at study initiation: (P) x wks; (F1) x wks:
( P) 7 to 8 weeks old
- Weight at study initiation: (P) Males: x-x g; Females: x-x g: Male 196.26 - 241.83 g,
Female 182.80-213.04 g
(F1) Males: x-x g; Females: x-x g: No data available
- Fasting period before study: A fast of 2h before chemical administered.
- Housing: Animals were housed four rats per sex per cage in sterilized solid bottom polypropylene cages with stainless steel grill tops with facilities for food, water bottles and bedding of clean paddy husk. The cages were suspended on stainless steel racks. Identified by assigned a unique identification (ID) number written on the tail, also specified on individual cage tag.
- Diet (e.g. ad libitum): Standard laboratory sterile extruded pelleted rodent feed, ad libitum.
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 30-70 %
- Air changes (per hr): 25 ± 5 air changes per hour.
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark

- Water (e.g. ad libitum): Potable tap water filtered through Reviva Reverse Osmosis
System (water filter cum purifier) of Eureka Forbes was provided in polypropylene bottles with stainless steel sipper tubes, ad libitum.
- Acclimation period: 5 days

Administration / exposure

Route of administration:
oral: gavage
Type of inhalation exposure (if applicable):
not specified
Vehicle:
corn oil
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: Dose of the test item was freshly prepared prior to dosing on each day. The test item Solvent blue 4 (SBL) was administered to each rat at the dose levels of 50, 150 or 450 mg/kg in the dose volume of 10 ml/kg body weight. The test item was weighed on a weighing balance. Then, it was transferred to calibrated falcon tube. Some quantity of the corn oil was added initially and vortexed. The sufficient quantity of vehicle was added to make up the required volume of dose.
Justification for use and choice of vehicle (if other than water): Corn oil was used as a vehicle for this study as the test item was not soluble in water and to deliver the desired dose levels because it is also recommended in the toxicological evaluation guidelines.
- Concentration in vehicle: 0, 50, 150 or 450 mg/kg body weight /day
Amount of vehicle (if gavage): 10 ml/kg body weight
- Lot/batch no. (if required): No data available
- Purity: No data available
Details on mating procedure:
No data available
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The concentration of the test item was calculated using the absorbance of the standard concentrations of the solvent blue.
Duration of treatment / exposure:
28 days
Frequency of treatment:
Daily
Doses / concentrations
Remarks:
Doses / Concentrations:
0, 50, 150 or 450 mg/kg/body weight/day
Basis:
nominal conc.
No. of animals per sex per dose:
Total : 56
0 mg/kg/body weight/day: 7 male, 7 female
50 mg/kg/body weight/day: 7 male, 7 female
150 mg/kg/body weight/day: 7 male, 7 female
450 mg/kg/body weight/day: 7 male, 7 female
Control animals:
yes, concurrent no treatment

Examinations

Parental animals: Observations and examinations:
Mortality and changes in body weight of the male and female reproductive organs like the testis, epidydimydes, uterus and ovary respectively were noted. In addition, food consumption, water consumption and locomotor activity effects were also recorded.
In male rats, relative organ weight of brain was significantly increased while no changes were observed in female organ weight when treated with 50 mg/kg/body weight/day as compare to control.
Estrous cyclicity (parental animals):
No changes detected as compared to control when treated with 50 mg/kg/day.
Sperm parameters (parental animals):
No changes detected as compared to control when treated with 50 mg/kg/day.
Litter observations:
No data available
Postmortem examinations (parental animals):
Testes, ovaries and uterus were pathologically examined after termination. Examinations also included weight of brain, epididymides, and ovaries (incl. paired ovaries and uterus, including cervix).
Postmortem examinations (offspring):
No data available
Statistics:
Statistical analysis was carried out by using Microsoft Excel and IBM SPSS statistics version- 20.0. All analyses and comparisons were evaluated at the 5 % level, statistically significant differences (p ≤ 0.05) indicated by appropriate notation. The focus was to examine the mean differences and their significance in control vs low-dose group only as there was total mortality observed in mid and high-dose groups in both the sexes. The statistical decision was taken by preparing the univariant GLM MODEL to check the significance between above mentioned groups. For multiple comparisons Turkey’s HSD test was applied.
Reproductive indices:
No data available
Offspring viability indices:
No data available

Results and discussion

Results: P0 (first parental animals)

General toxicity (P0)

Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
When treated with 450 mg/kg/body weight/day, all the male and female were dead on day 5 as compare to control. When treated with 150 mg/kg/body weight/day, all the male and female were dead on day 11 as compare to control.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
When treated with 50 mg/kg/body weight/day significant decrease in body weight was observed in treated male and female rat as compare to control.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
When treated with 50 mg/kg/body weight/day significant decrease in body weight was observed in treated male and female rat as compare to control.
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
In male rats, relative organ weight of brain was significantly increased while no changes were observed in female organ weight when treated with 50 mg/kg/body weight/day as compare to control
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
When treated with 50 mg/kg/body weight/day perineum stained bluish in color, alimentary canal including stomach and intestine stained bluish in color in male and female rat, Mild blue tinged patches and stomach blotted with gastric content were observed
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
No significant changes were observed in the reproductive organs of 50 mg/kg/body weight/day treated male and female rat as compared to control.
Other effects:
not specified

Reproductive function / performance (P0)

Reproductive function: estrous cycle:
effects observed, treatment-related
Description (incidence and severity):
No changes detected as compared to control when treated with 50 mg/kg/day.
Reproductive function: sperm measures:
effects observed, treatment-related
Description (incidence and severity):
No changes detected as compared to control when treated with 50 mg/kg/day.
Reproductive performance:
not specified

Details on results (P0)

Male and female rats dosed with 50 mg/kg bw of Solvent blue 4 did not show appreciable difference in the weight of testes & epidydimides and uterus and ovaries respectively, as compared to the control. No significance attributed to the reproductive organ body weight changes in comparison to control groups.

Effect levels (P0)

Dose descriptor:
NOAEL
Effect level:
50 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: based on observations of reproductive organs like testes, ovaries and uterus that were pathologically examined after termination

Results: F1 generation

General toxicity (F1)

Clinical signs:
not specified
Mortality / viability:
not specified
Body weight and weight changes:
not specified
Sexual maturation:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
not specified

Details on results (F1)

Details not available

Overall reproductive toxicity

Reproductive effects observed:
not specified

Any other information on results incl. tables

Summary of Reproductive Organ Weights (Male Rats)

Group/Dose

 

Testes

Epididymides

1/0 mg/kg bw

Mean

0.9930

0.4157

SD

0.1199

0.0675

SEM

0.0453

0.0255

2/50 mg/kg bw

Mean

1.0826

0.4780

SD

0.1111

0.0530

SEM

0.0420

0.0200

No significance attributed to the body weight changes in comparison to control groups

Summary of Reproductive Organ Weights (Female Rats)

Group/Dose

 

Uterus

Ovaries

5/0 mg/kg bw

Mean

0.2915

0.0441

SD

0.0519

0.0188

SEM

0.0196

0.0071

6/50 mg/kg bw

Mean

0.2346

0.0412

SD

0.0701

0.0155

SEM

0.0265

0.0059

No significance attributed to the body weight changes in comparison to control groups

Applicant's summary and conclusion

Conclusions:
Male and female rats dosed with 50 mg/kg bw of Solvent blue 4 did not show appreciable difference in the weight of testes & epidydimides and uterus and ovaries respectively, as compared to the control. No significance attributed to the reproductive organ body weight changes in comparison to control groups.
Hence the no observed adverse effect level (NOAEL) for the parents has been concluded to be 50 mg/kg bw
Executive summary:

In a 28 days repeated dose toxicity study, the effect on the reproductive organs of Solvent blue 4 (containing less than 0.1% Michler's Ketone) was evaluated in male and female Sprague Dawley rats. The test chemical was administered by oral gavage in the concentration of 0,50, 150 or 450 mg/kg body weight/day. The results showed that Solvent blue 4in treated rats had nasal discharge, red crust around nostrils and soft feces and vocalization on handling, and decreased body weight were also observed as compare to control. In male rat, there were increased levels of RBC, monocytes, potassium and total protein, while female rats showed decreased level of total bile acid as compare to control.

Histopathology showed no significant changes at treatment with 50 mg/kg/body weight/day as compare to control. In addition, male and female rats dosed with 50 mg/kg body weight of Solvent blue 4 did not show appreciable difference in the weight of testes and epidydimides or in uterus and ovaries as compared to the control. Therefore, NOAEL is considered to be 50 mg/kg/day when male and female Sprague Dawley rats were exposed daily toSolvent blue 4 by oral route for 28 days.