Petroleum kerosene fraction, co-processed with renewable hydrocarbons of plant and/or animal origin

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Effect on fertility: via oral route

Endpoint conclusion:

no study available

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

494 mg/kg bw/day

Additional information

Data on related substances have been used to 'read-across' and predict the hazard properties. A 'read-across' justification document can be found in section 13.

In a read-across reproductive/developmental toxicity screening study (Klimisch score=1; Schreiner et al., 1997), 10 Sprague Dawley rats/sex/group were treated dermally with hydrodesulfurised kerosine at concentrations of 0 (sham-treated and vehicle control groups), 165, 330 or 494 mg/kg/day (0, 20, 40 or 60% (v/v) respectively) in mineral oil in a dosing volume of 1 mL/kg for a minimum of 6 hours, 7 days/week beginning 14 days premating, during the 14-day mating period and through 20 days of gestation. Rats were mated overnight on a 1:1 ratio and were separated the following morning. 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 occurred in both males (all doses) and females (high dose only).

At terminal sacrifice, no findings were reported except for those on the skin. 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. No effects were observed on reproductive function or outcome.

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.

 

Short description of key information:

Kerosine does not cause fertility effects (OECD 421).

Effects on developmental toxicity

Description of key information

Two relevant studies were identified. In the key dermal study with hydrodesulphurised kerosine, no developmental effects were observed at levels upto 494 mg/kg/day. In a supporting study exposure of rats to kerosine vapour at levels upto 364 ppm did not cause any developmental effects in progeny.

Link to relevant study records

Reference

Reference 1

Endpoint:

developmental toxicity

Type of information:

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

Adequacy of study:

supporting 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 410.

Qualifier:

according to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Deviations:

yes

GLP compliance:

not specified

Species:

rat

Strain:

Sprague-Dawley

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Breeding Laboratories, Inc., Wilmington, Massachusetts
- Age at study initiation: 12 weeks old
- Weight at study initiation: Not reported
- Housing: Individually in wire cages
- Diet (e.g. ad libitum): ad libitum except during exposure period
- Water (e.g. ad libitum): Not reported
- Acclimation period:13 days

Route of administration:

inhalation

Vehicle:

air

Details on exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: Dynamic 0.25 cubic metre stainless steel and plexiglass chamber operated under negative pressure
- Method of holding animals in test chamber: Stainless steel and plexiglass chamber
- System of generating vapour: Metering it into a warmed flask and passing compressed air through the flask , then diluted with room air as it entered the chamber
- Temperature, humidity, pressure in air chamber: 21.1 to 23.7 degrees Celsius, humidity not reported, chamber air flow rate was 28.3 litres/minute
- Air flow rate: Not reported
- Air change rate: Not reported


TEST ATMOSPHERE
- Brief description of analytical method used: Hourly samples were analyzed using a Scott Model 216 Hydrocarbon Analyzer previously calibrated with known concentrations of kerosine with methane used as an internal standard
- Samples taken from breathing zone: no

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Intended exposure concentrations were 0, 100, and 400 ppm, 66 to 73 hourly samples were obtained over the course of the exposure period, the average concentrations were 0, 106.4 +/- 10.23, and 364.0 +/- 37.53 ppm, respectively.

Details on mating procedure:

- Impregnation procedure: cohoused
- If co-housed:
- M/F ratio per cage: 1 male to 1 female
- Length of cohabitation: Not reported
- Proof of pregnancy: vaginal plug referred to as day 0 of pregnancy

Duration of treatment / exposure:

Six hours each day

Frequency of treatment:

Daily

Duration of test:

10 days from gestational day 6 through gestational day 15

Remarks:

Doses / Concentrations:
106 or 364 ppm
Basis:
analytical conc.

No. of animals per sex per dose:

20 pregnant females

Control animals:

yes

Details on study design:

- Dose selection rationale: Only stated that doses were selected by the American Petroleum Institute

Maternal examinations:

CAGE SIDE OBSERVATIONS: No data


DETAILED CLINICAL OBSERVATIONS: No data


BODY WEIGHT: Yes
- Time schedule for examinations: Gestational days 0, 6, 15, and 20.


FOOD CONSUMPTION: Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data


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



POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 20
- Organs examined: Visceral and thoracic organs

Ovaries and uterine content:

The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: No
- Number of corpora lutea: No
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
- Other: It was only stated that the number of resportion sites were reported. The placement of the implantation sites in the uterine horns were replicated. The number of live and dead fetuses were recorded.

Fetal examinations:

- External examinations: Yes: all per litter
- Soft tissue examinations: Yes: one-third of the fetuses
- Skeletal examinations: Yes: two thirds of the fetuses
- Head examinations: Yes: one-third of the fetuses (the same ones that had soft tissue examinations)

Statistics:

The litter was used as a basic sampling unit for statistical analysis. Dunnett's t-test was used to determine statistical significance (P<0.05) with regard to differences between means with near-normal distribution (body weights and food consumption of dams, mean pup weight based on litter averages). Ratios (nidation index and implantation/corpora lutea ratio) were analyzed with a 2 x 2 contingency table with Yates' correction. Wilcoxon Rank Sum was used for discontinuous parameters as measured by the number of abnormal foetuses within a litter.

Details on maternal toxic effects:

Maternal toxic effects:no effects

Dose descriptor:

NOAEC

Effect level:

>= 364 ppm

Basis for effect level:

other: maternal toxicity

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Dose descriptor:

NOAEC

Effect level:

>= 364 ppm

Basis for effect level:

other: teratogenicity

Abnormalities:

not specified

Developmental effects observed:

not specified

There were no compound-related deaths or clinical signs throughout the study. At necropsy two animals in the 100 ppm group had lung mottling, but this was considered to be an incidental finding. There were no significant differences in either body weight or food consumption data. The following is a summary of reproductive data base on observations of the uterine contents on day 20 of gestation

Dose (ppm)
Observation	Historical control*	0	100	400
Nidation index (females with implantations/Bred)
	55/61 (90.2%)	19/20 (95%)	18/20 (90%)	18/20 (90%)
Females dying prior to Caesarean section
	0	0	0	0
Live litters	54	19	18	18
Implantation sites (left horn/right horn)
	301/363	110/118	106/138	126/114
Resorptions
Total	43	18	21	12
Litters	24	11	10	7
Dead foetuses
Total	0	0	0	0
Litters	0	0	0	0
Mean live litter size (foetuses)
	11.3	11.1	12.4	12.7
Average foetal weight (g)
	4.1	4.3	4.1	3.9
*       Based on 54 litters

Examination of offspring at delivery did not reveal any treatment-related abnormalities.
Examination of Bouin's fixed specimens did not reveal any treatment-related abnormalities.
The sex ratio was also unaffected by treatment.

Skeletal examinations revealed the following:

Dose group	No. foetuses examined (number of litters)	No. Foetuses with normal	Foetuses
			common* changes	unusual skeletal
			variations
0	140 (19)	60	72 (16)	8 (4)
106.4	150 (18)	69	72 (15)	9 (4)
364	154 (18)	62	84 (18)	8 (5)

* Foetuses with commonly-encountered changes only. The authors comment that most of the changes, while not strictly normal, are frequently observed in 20-day-old rat foetuses of this strain and source in their laboratory. The changes were not malformations, but were mostly related to retarded bone ossification. Neither the frequency nor the character of the changes indicated an adverse effect on foetal growth and development or a teratogenic potential.

Conclusions:

The NOAEC for both maternal and developmental toxicity was 364 ppm kerosine.

Executive summary:

In a pre-natal developmental study, 20 female presumed-pregnant rats per group were exposed to 106 or 364 ppm kerosine vapour. Exposures were accomplished in chambers for 6 hours each day on days 6 through 15 of gestation. A control group of 20 presumed-pregnant rats of the same age served as controls and were placed in chambers and exposed to room air only. The rats were weighed on days 0, 6, 15 and 20 of gestation and food consumption was measured throughout the study. The animals were also observed daily for clinical signs. On day 20 of gestation, the animals were anesthetized with chloroform and the visceral and thoracic organs were examined. The uterus was removed and opened and the number of implantation sites and their placement in the uterine horns were recorded. Live and dead foetuses and resorption sites were also recorded and the foetuses were removed, examined externally for abnormalities and weighed. One third of the foetuses was fixed in Bouin's fluid and were examined subsequently for changes in the soft tissues of the head, thoracic and visceral organs. The remaining foetuses were stained with Alizarin red S and examined subsequently for skeletal abnormalities.

There were no effects noted on either the dams of the foetuses. The NOAEC is greater than or equal to 364 ppm, based on the lack of effects and no LOAEC can be determined.

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 410.

Reference 2

Endpoint:

developmental toxicity

Type of information:

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

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.

Qualifier:

equivalent or similar to guideline

Guideline:

EPA OTS 798.4900 (Prenatal Developmental Toxicity Study)

GLP compliance:

not specified

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Details on test animals or test system and environmental conditions:

TEST ANIMALS- Source: Charles River Breeding Laboratories, Kingston, New York-
Age at study initiation: Females were approximately 8 weeks old
Weight at study initiation: 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

Route of administration:

dermal

Vehicle:

other: 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.

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.

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

Duration of treatment / exposure:

Exposure period: 14 days premating to day 20 of gestation.

Frequency of treatment:

Daily

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.

Maternal 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: 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

Details on maternal toxic effects:

Maternal toxic effects:yes. Remark: skin irritation only

Details on maternal toxic effects:
The only signs of maternal toxicity were skin irritaaion in the highest dose level

Key result

Dose descriptor:

NOAEL

Effect level:

>= 494 mg/kg bw/day

Basis for effect level:

other: maternal toxicity

Remarks on result:

other: The only signs of maternal toxicity were skin irritaaion in the highest dose level

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Dose descriptor:

NOAEL

Effect level:

>= 494 mg/kg bw/day

Basis for effect level:

other: developmental toxicity

Remarks on result:

other: no developmental effects

Abnormalities:

not specified

Developmental effects observed:

not specified

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	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	2	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	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.

Conclusions:

Dermal application of hydrodesulphurised kerosine did not cause any developmental effects on progeny.

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 494mg/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.

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEC

364

Species:

rat

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

494 mg/kg bw/day

Additional information

In a key reproductive/developmental toxicity screening study (Klimisch score=1; Schreiner, et al., 1997), 10 Sprague Dawley rats/sex/group were treated dermally with hydrodesulfurised kerosine at concentrations of 0 (sham-treated and vehicle control groups), 165, 330 or 494 mg/kg/day (0, 20, 40 or 60% (v/v), respectively) in mineral oil in a dosing volume of 1 ml/kg for a minimum of 6 hours,7 days/week beginning 14 days premating, during the 14-day mating period and through 20 days of gestation. Dosage equivalents were 0, 165, 330 and 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 was the highest dose tested in the study.

  

In a supporting pre-natal developmental study (Klimisch score=1; API, 1979b), 20 female presumed-pregnant rats per group were exposed to 106 or 364 ppm kerosine vapour for 6 hours each day on days 6 through 15 of gestation. A control group of 20 presumed-pregnant rats of the same age served as controls and were placed in chambers and exposed to room air only. The animals were sacrificed on day 20 of gestation. There were no effects noted on either the dams or the foetuses. The NOAEC is greater than or equal to 364 ppm, based on the lack of effects and no LOAEC can be determined.

 

Justification for selection of Effect on developmental toxicity: via inhalation route:

well conducted inhalation study in rats on kerosine

Justification for classification or non-classification

Available data indicate that kerosine has no effect on the fertility of male and female rats and does not cause developmental effects. Therefore kerosines are not classified for reproductive or developmental toxicity according to EU CLP Regulation (EC No. 1272/2008).