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Toxicological information

Toxicity to reproduction

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Administrative data

Endpoint:
screening for reproductive / developmental toxicity
Remarks:
based on test type (migrated information)
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: This study is classified as reliable without restrictions because it was carried out in a method equivalent/similar to OECD TG 421.

Data source

Reference
Reference Type:
publication
Title:
Toxicity evaluation of petroleum blending streams: reproductive and developmental effects of hydrodesulfurized kerosine
Author:
Schreiner, C., Bui, Q., Breglia, R., Burnett, D., Koschier, F., Podhasky, P., Lapadula, L., White, R., Feuston, M., Krueger, A., Rodriguez, S.
Year:
1997
Bibliographic source:
Journal of Toxicology and Environmental Health 52:211-229

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
GLP compliance:
not specified

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Hydrodesulfurised kerosine, CAS No. 64742-81-0
- Other:
The chemical composition of the sample of hydrodesulfurised
kerosine was determined by ASTM method D 1319-1 and the
results are tabulated below.

Component Weight %
Nonaromatics 80.27
Saturates 78.61
Olefins 1.66
Aromatics 19.72
<3-ring PAC >19.72
3-7 -ring PAC <0.01

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Breeding Laboratories, Kingston, New York
- Age at study initiation: Males and females were approximately 8 weeks old
- Weight at study initiation: (P) Males: 275 to 285 grams; Females: 183 to 187 grams
- Use of restrainers for preventing ingestion (if dermal): yes
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 12 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 to 22
- Humidity (%): 40 to 60%
- Photoperiod (hrs dark / hrs light): 12 hours dark/12 hours light


Administration / exposure

Route of administration:
dermal
Vehicle:
other: Squibb mineral oil (340 SUS)
Details on exposure:
TEST SITE
- Area of exposure: Entire dorsal back
- Type of wrap if used: None
- Time intervals for shavings or clippings: Once a week


REMOVAL OF TEST SUBSTANCE
- Washing (if done): Washing was only stated to have been done during mating and consisted of wiping the area clean with a piece of gauze.
- Time after start of exposure: A minimum of 6 hours


TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 1 ml/kg body weight/day
- Concentration (if solution): 0, 20%, 40%, or 60%
- Constant volume or concentration used: yes


VEHICLE
- Justification for use and choice of vehicle (if other than water): Vehicle was used to reduce skin irritation without compromising dermal absorption.
- Amount(s) applied (volume or weight with unit): 1 ml/kg body weight/day

USE OF RESTRAINERS FOR PREVENTING INGESTION: yes, Elizabethan collars were used.
Details on mating procedure:
- M/F ratio per cage: 1 to 1
- Length of cohabitation: Overnight
- Proof of pregnancy: Vaginal plug or sperm in vaginal lavage sample referred to as day 0 of pregnancy
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Details only specify that all test solutions were within +/- 5.42% of the original calculated concentration or of the nominal concentration.
Duration of treatment / exposure:
Exposure period: 14 days premating to day 20 of gestation with males treated for an additional week
Premating exposure period (males): 14 days
Premating exposure period (females): 14 days
Frequency of treatment:
Daily, 7 days/week
Doses / concentrations
Remarks:
Doses / Concentrations:
165 (20%), 330 (40%) & 494 (60%) mg/kg/day
Basis:
other: Different concentrations in solution and amount applied
No. of animals per sex per dose:
Ten
Control animals:
yes
Details on study design:
- Dose selection rationale: Doses were selected based on a two-week range finding study.

Examinations

Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: No


DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Twice a day, but once a day on weekends and holidays


BODY WEIGHT: Yes
- Time schedule for examinations: Males were weighed on the first day of dosing, weekly thereafter, and at study termination. Females were weighed on the first day of dosing, weekly thereafter until mating was confirmed, then on gestational days 0, 3, 6, 10, 13, 16, and 20, on postpartum day 0 and 4, and at terminal sacrifice.


FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No


WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No

Sperm parameters (parental animals):
Parameters examined in P male parental generations: testis weight
Litter observations:
STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: No


PARAMETERS EXAMINED
The following parameters were examined in F1 offspring: number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, weight gain


GROSS EXAMINATION OF DEAD PUPS: Yes, for external abnormalities; possible cause of death was not determined for pups born or found dead.
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: All surviving animals were sacrificed after all the females had been sacrificed.
- Maternal animals: Surviving animals that did not deliver were sacrificed on gestation day 25. Dams that delivered were sacrificed on postpartum day 4 to postpartum day 6.


GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations with emphasis on the reproductive organs.


HISTOPATHOLOGY / ORGAN WEIGHTS
The liver, kidneys, adrenals, thymus, spleen, brain and heart of all parental animals were weighed. In addition the testes and epididymides of parental males were weighed. Skin from treated sites, ovaries and testes and epididymides were prepared for histological examination. Pathological evaluation was performed on reproductive organs from all males and pregnant females in both control groups and the high dose group and on treated skin from all groups.
Postmortem examinations (offspring):
No tissues from offspring were retained.
Statistics:
Quantitative data (body weight and food consumption) were analyzed by parametric methods: analysis of variance (ANOVA) and associated F-test, followed by Dunnet's test for multiple comparisons, provided there was statistical significance in the ANOVA. Maternal reproductive data were evaluated by ANOVA followed by group comparisons using Fisher's exact test. Differences between control and treatment groups were considered statistically significant only if the probability of the differences being due to chance was less than 5% (P< 0.05).
Reproductive indices:
Fertility index, corpora lutea, implantation sites, litters with live born pups
Offspring viability indices:
Number of pups delivered, live birth index, pup mortality, pups surviving to postnatal day 4 (viability index), pup weight

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

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
Other effects:
not specified

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
not specified
Reproductive function: sperm measures:
not specified
Reproductive performance:
no effects observed

Details on results (P0)

CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS): There were no treatment-related effects on clinical signs or mortality.


BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS): There were no significant effects on body weight, but high-dose males had slightly (<10%) lower body weight gain compared to the controls.


REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS): There were no treatment-related effects on reproductive performance.


ORGAN WEIGHTS (PARENTAL ANIMALS): There was a significant increase in the relative kidney weight in high-dose males compared to the sham control that was likely related to the slightly lower terminal body weight in this group. There were no treatment-related changes in reproductive organ weights.


GROSS PATHOLOGY (PARENTAL ANIMALS): Skin irritation was the only effect observed.


HISTOPATHOLOGY (PARENTAL ANIMALS): There were no treatment-related changes in histopathology.


OTHER FINDINGS (PARENTAL ANIMALS): Mild to moderate skin irritation was observed in both males and females. There was a dose-dependent increase in skin irritation in the males, but skin irritation was only observed in high-dose females. The vehicle (mineral oil) caused a slight increase in irritation in both males and females. Histopathology confirmed the dermal irritation.

Effect levels (P0)

Dose descriptor:
NOAEL
Remarks:
reproductive toxicity
Effect level:
>= 494 mg/kg bw/day
Sex:
male/female

Results: F1 generation

General toxicity (F1)

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

Effect levels (F1)

Dose descriptor:
NOAEL
Remarks:
developmental (offspring) toxicity
Generation:
F1
Effect level:
>= 494 mg/kg bw/day
Sex:
male/female

Overall reproductive toxicity

Reproductive effects observed:
not specified

Any other information on results incl. tables

One pregnant mid-dose female died before delivery. No other animals died or were prematurely sacrificed and no clinical signs of toxicity were observed.
Skin irritation among males varied from slight to moderate with increasing dose and was most severe in the high dose group.
 Mild to moderate skin irritation was observed in females at the highest concentration.
At terminal sacrifice, no findings were reported except for those on the skin. Microscopic changes were found in the skin of males in the vehicle control and all kerosine-treated groups. In females changes were only observed in the high dose group animals. The skin findings (macroscopic and microscopic) are shown in the following table.

 

 

 Kerosine (mg/kg)

 

Parameter 

Control

Mineral oil

165

330

494

Males

 

 

 

 

 

No animals

10

10

  10

 10

 10

Max. skin irritation score, sum of means

 

 

 

 

 

Week of max severity

-

 2

 5

 5

Mean (SD)

 0

1.3(1.2)

2.4(0.7)

2.5(2.0)

3.3(2.1)

Min/max score

0

0/3

 1/3

 0/7

  1/7

Gross necropsy observations

 

 

 

 

 

Crust/scab

1

0

  0

 0

   1

Scaly/dry/flaky

0

  0

 1

   2

 3

Histopathological observations

 

 

 

 

 

Acanthosis/hyperkeratosis

 2

 5

 8

 7

 8

Hyperplasia, sebaceous glands

 3

 5

 5

 3

 5

Inflammation, dermal

 1

 6

 6

 7

Necrosis, epidermal, focal

 1

 0

 1

 1

 5

Females

 

 

 

 

 

No animals

10

6

  10

 10

 10

Max. skin irritation score, sum of means

 

 

 

 

 

Week of max severity

 6

 7

 3

 4

 4

Mean (SD)   

 0.2 (0.6)

0.7 (1.0) 

 0.4 (0.8)

 1.1 (0.9)

 2.3 (1.8)

Min/max score

0/2

0/2

 0/2

 0/2

  1/7

Gross necropsy observations

 

 

 

 

 

Crust/scab

0

1

  1

 0

   3

Scaly/dry/flaky

0

  0

 0

   0

 0

Histopathological observations

 

 

 

 

 

Acanthosis/hyperkeratosis

 3

 2

 5

 5

 6

Hyperplasia, sebaceous glands

 1

 0

 0

 0

 1

Inflammation, dermal

 0

 1

 1

 1

 4

Necrosis, epidermal, focal

 0

 0

 0

 0

 0


Body weights were unaffected by treatment. However over the course of the 8 weeks, high dose males gained less weight than the controls (201 g compared to 237g for the sham controls). Food consumption was unaffected by treatment. High dose males had a higher mean relative kidney weight than sham controls (0.76 vs 0.66).
 This was attributed to the lower mean final body weights of the high dose group. No other organ or organ/body weight changes were recorded.

Controls

Kerosine (mg/kg)

Parameter

Sham

Oil

165

330

494

No animals

10

10

10

10

10

Fertility index

100%

90%

90%

80%

100%

Litter with liveborn pups

 10

 9

 7a

 10

Corpora lutea 

 

 

 

 

 

Number

      169

151

158

122

172

Mean

16.9

16.8

17.6

17.4

17.2

(SD)

       (1.9)

(2.4)

(2.0)

(0.8)

(2.9)

Implantation sites

 

 

 

 

 

Number

      163

149

155

118

167

Mean

      16.3

16.6

17.2

16.9

16.7

(SD)

      (1.9)

(2.4)

(1.8)

(1.3)

(2.8)

Pups delivered

 

 

 

 

 

Total

      152

131

147

109

150

Mean

      15.2

14.6

16.3

15.6

15.0

(SD)

      (2.0)

(2.7)

(2.3)

(2.9)

(2.9)

Liveborn

152

130

143

108

148

Livebirth index

100%

99%

97%

99%

99%

Pups dying

 

 

 

 

 

day 0

      3

0

1

1

1

days 1-4

2

4

1

1

9bc

Pups surviving

 

 

 

 

 

4 days

       147

126

141

106

138

Viability index

97

97

99

98

93c

Pup weight/litter (g)

 

 

 

 

 

day 1 mean

6.9

6.8

7.0

7.0

6.7

day 4 mean

9.9

9.6

10.1

9.9

9.8 

a       one dam died on gestation day 21; the cause of death was unrelated to treatment
b
       significantly different from control (P<0.05)
c
       One dam had a malfunctioning water bottle; when 4 dead pups from this litter are excluded from the analysis, no
significant difference from control was found.

No test-material-related microscopic changes were observed in the testes or epididymides of adult male rats or in the ovaries of adult female rats.

Applicant's summary and conclusion

Conclusions:
The test compound did not cause any reproductive or developmental toxicity.
Executive summary:

In a reproductive/developmental toxicity screening study, ten male Sprague Dawley rats (aged approximately eight week old, weighing 275-285g) and 10 females (same age and weighing 183-187g) were treated dermally with hydrodesulfurised kerosine at concentrations of 0 (sham-treated and vehicle control groups), 20, 40 or 60% (v/v) in mineral oil in a dosing volume of 1 mL/kg. Dosage equivalents were 0, 165, 330 and 494 mg/kg/day. Test material was applied daily to the shorn skin of the animals 7 days/week beginning 14 days premating, during the 14 day mating period and through 20 days of gestation. Males were treated for an additional week. Collars were fitted to the animals during the dosing period to prevent ingestion of applied materials, but the test site was not covered. After the final dose, the collars were removed and residual test material was wiped from the skin. There were two control groups: the vehicle control was given mineral oil only and in the sham-treated group the animals had been fitted with collars and were stroked with the tip of a syringe, but no material was applied.

During the mating period the test material remained on the animal's backs for at least 6 hours. Prior to pairing, the test material was removed by wiping. Rats were mated overnight on a 1:1 ratio and were separated the following morning. Collars were then reapplied prior to the next dose being applied. Females were monitored for evidence that mating had taken place. Pregnancy was determined by the presence of a vaginal plug or sperm in a vaginal lavage sample. If observed, the female was considered to be at day 0 of gestation. Any female that did not show evidence of mating was placed with the same male the following evening. Any female that did not show evidence of mating at the end of a 2 week mating period was presumed pregnant (gestation day 0 = last day of cohabitation).

Skin irritation among males varied from slight to moderate with increasing dose and was most severe in the high-dose group. Mild to moderate skin irritation was observed in females at the highest concentration. At terminal sacrifice, no findings were reported except for those on the skin. Microscopic changes were found in the skin of males in the vehicle control and all kerosine-treated groups. In females changes were only observed in the high-dose group.

Body weights were unaffected by treatment. However over the course of the 8 weeks, high-dose males gained less weight than the controls. Food consumption was unaffected by treatment. High-dose males had a higher mean relative kidney weight than controls (0.76 vs 0.66). This was attributed to the lower mean final body weights of the high-dose group. No other organ or organ/body weight changes were recorded.

No test-material-related microscopic changes were observed in the testes or epididymides of adult male rats or in the ovaries of adult female rats.

There is no parental systemic LOAEL, based on the lack of any significant treatment-related findings except dermal irritation. The parental systemic NOAEL is greater than or equal to 494 mg/kg/day.

 

There is no offspring LOAEL, based on the lack of any effects noted in the offspring. The offspring NOAEL is greater than or equal to 494 mg/kg/day.

 

This study received a Klimisch score of 1 and is classified as reliable without restrictions because it was carried out in a method equivalent/similar to OECD TG 421.