Decene, hydroformylation products, low boiling

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Link to relevant study records

Reference

Reference 1

Endpoint:

fertility, other

Remarks:

based on test type (migrated information)

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

1991

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Acceptable well-documented study report which meets basic scientific principles.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 415 [One-Generation Reproduction Toxicity Study (before 9 October 2017)]

Principles of method if other than guideline:

Female rats were dosed with neat JP-8 (0, 325, 750, 1500 mg/kg) daily by gavage for a total of 21 weeks (90-day plus mating with naive males, gestation and lactation) in an effort to assess general toxicity, fertility and reproductive endpoints.

GLP compliance:

not specified

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Breeding Labs, Kingston, NY
- Weight at study initiation: (P) Females: 180 to 200 g
- Diet (e.g. ad libitum): Formula 5008, Ralston Purina, St. Louis, MO, ad libitum
- Water (e.g. ad libitum):ad libitum
- Acclimation period: 2 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21-25
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on exposure:

JP-8 was administered by gavage without a vehicle (neat). Control animals were dosed with 1.0 mL distilled water under the same conditions as test groups. Volumes to be administered each day were calculated from the individual daily body weights and the density of the test material (0.81 g/mL).

Details on mating procedure:

The rats were given 0 (control), 325, 750 or 1500 mg/kg JP-8 daily by gavage for 21 weeks (90-days followed by cohabitation, gestation, delivery and lactation). The male rats, not exposed to JP-8, were housed 1:1 with treated female rats. Dams were euthanized one day after weaning (Day 22 of lactation); male rats were euthanized after pregnancy was confirmed. Litters were standardized to four male pups and four female pups on postnatal day (PND) 5. All rats were euthanized by carbon dioxide overdose.

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

90 days followed by cohabitation, gestation, delivery and lactation

Frequency of treatment:

daily; 7 days/ week

Details on study schedule:

Young female Sprague-Dawley rats weighing 180-200 g were randomly assigned to 4 exposure groups. Groups contained a minimum of 35 female rats. The rats were given 0 (control), 325, 750 or 1500 mg/kg JP-8 daily by gavage for 21 weeks (90-days followed by cohabitation, gestation, delivery and lactation). The male rats, not exposed to JP-8, were housed 1:1 with treated female rats. Dams were euthanized one day after weaning (Day 22 of lactation); male rats were euthanized after pregnancy was confirmed. Litters were standardized to four male pups and four female pups on postnatal day (PND) 5. All rats were euthanized by carbon dioxide overdose.

Remarks:

Doses / Concentrations:
0 (control), 325, 750 or 1500 mg/kg
Basis:
actual ingested

No. of animals per sex per dose:

35 females

Control animals:

yes, sham-exposed

Positive control:

none

Parental animals: Observations and examinations:

A subset of dams from each treatment group (maximum n of 10) was selected for hematology, clinical chemistry and urine analyses. The same subsets were also used for organ weights and histopathology. Whole blood was collected from the dam's inferior vena cava at necropsy. The following hematology parameters were measured: red blood cell count, hemoglobin, mean corpuscular volume, mean corpuscular hemoglobin, red cell distribution width, mean corpuscular hemoglobin concentration, hematocrit, platelet count and differential leukocyte count. Determinations were made using an automated counter (H-1 System, Technicon Instruments, Corporation, Tarrytown, NY).

The following clinical chemistry parameters were measured in serum from dams: sodium,
glucose, magnesium, carbon dioxide, potassium, albumin, chloride, total protein, calcium, blood urea nitrogen, total bilirubin, uric acid, inorganic phosphate, creatinine, triglycerides, cholesterol, AST, ALT, alkaline phosphatase, lactate dehydrogenase, creatine kinase and gamma-glutamyl transferase. Assays were performed with the Ektachem 700XR chemistry analyzer (Eastman Kodak, Rochester, NY).

The dams were transferred to metabolism cages upon weaning and urine was collected for 24- hours prior to sacrifice. The total volume of urine was noted and the urine was assayed for pH, specific gravity, total protein and creatinine. Specific gravity was measured with a refractometer (American Optical Corp., Southbridge, MA) and pH was measured with an electronic meter (Model 601A, Orion Research Inc., Cambridge, MA). Urine protein assays were performed on an automated chemistry analyzer (Model ACA IV, DuPont, Wilmington, DE). Urine creatinine determinations were made using the Ektachem 700XR analyzer.

Oestrous cyclicity (parental animals):

not examined

Sperm parameters (parental animals):

not examined

Litter observations:

not examined

Postmortem examinations (parental animals):

A gross pathologic examination was performed on a subset of dams from each treatment group following euthanasia. A maximum of 10 rats per group was designated for clinical pathology and histopathology. Tissues were collected and prepared for histopathologic examination and included: gross lesions, thymus, brain, kidneys, lungs, adrenals, trachea, pancreas, heart, ovaries, uterus, liver, nasal turbinates, spleen, esophagus, duodenum, stomach, jejunum, colon, ileum, rectum, urinary bladder, sternum, mandibular lymph nodes, sciatic nerve, mesenteric lymph nodes, skeletal muscle. Brain, kidneys, liver, spleen and ovaries were weighed during necropsy.

Postmortem examinations (offspring):

not examined

Statistics:

Statistical Analyses for General Toxicity Data
Adult body weights, organ weights and urine metabolites were analyzed by an ANOVA with multiple comparisons. Clinical chemistry results, hematology values, urinalysis data and severity of pathological changes were compared using an ANOVA. The level of significance was accepted at p<0.05 unless stated otherwise.

Statistical Analyses for Reproductive Measures
A one-factor (dose) or two-factor (dose and pup sex) analysis of variance (ANOVA) was performed for continuous variables. Error terms used were either dam(dose) or pup sex and dam(dose). One-way ANOVA was used with gestation lengths, sperm parameters and litter sizes while two-way was used for pup weights. Post-hoc paired comparisons of dose used two-tailed t-tests with pooled error.

For categorical variables, a Chi-square test of proportions was used to determine differences among the doses. Chi-square tests were used for pregnancy rates and percent viability. Posthoc paired comparison for the viability parameter was performed with Fisher's Exact test. The level of significance was accepted at p<0.05 unless stated otherwise.

Reproductive indices:

Gestation day 0 was determined by presence of copulatory plug or sperm in a vaginal contents smear. Pregnancy rate (%) and gestation duration (days) were recorded for all dams. Size of the entire litter and the number born dead were noted on PND 1; the resulting numbers were compared between treatment groups.

Offspring viability indices:

Size of the entire litter and the number born dead were noted on PND 1; the resulting numbers were compared between treatment groups. Pups were weighed on PND 1, 4, 7, 14, 21 and 90; male and female pup weights were compared between treatment groups and between sexes.

Clinical signs:

no effects observed

Body weight and weight changes:

effects observed, treatment-related

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Other effects:

not examined

Reproductive function: oestrous cycle:

not examined

Reproductive function: sperm measures:

not examined

Reproductive performance:

no effects observed

Results of the 21-week oral gavage exposure to 0, 325, 750 and 1500 mg/kg/day JP-8 revealed a decrease in body weights of the female rats. Body weights for the 1500 mg/kg/day rats were significantly lower than control rats (p<0.01) starting at week 8 and continuing throughout gestation and most of lactation (weeks 13-20). Terminal body weights at week 21 were not significantly different from control rat weights. Mortality in each treatment group was not related to dose.

Pregnancy rates and litter sizes for treated animals were not significantly different from controls. Gestation length was calculated from dams that became pregnant within one estrous cycle. Of the 87 dams that became pregnant, 77 took 1 to 4 days of cohabitation to become pregnant. The remaining 13 dams had reported times to impregnation ranging from 5 to 11 days. Most of these dams had gestation lengths as short as 14 days due to misidentification of the first day of impregnation. Therefore, time to impregnation plus gestation length was determined for each group. Since there were no significant differences between control and exposure groups for the combined impregnation/gestation length, dams with long impregnation times and short gestation lengths were excluded from the gestation length calculation. There were still no significant differences seen in gestation length for the remaining dams. The percentage of live pups on Day 1 for each dose group was not different from the control percent. The number of dams per treatment with at least one dead pup was not different between the dose groups and control.

No significant changes were found in urine parameters (total volume, specific gravity and creatinine concentration). There were no statistically significant changes in most hematology counts: neutrophil, eosinophil, basophil, lymphocyte and platelet. The leukocyte count was found to be significantly decreased in the 325 mg/kg/day dose group alone (p<0.05). This change was not dose dependent and has questionable biological significance. Clinical chemistry values (sodium, chloride, glucose, triglycerides, creatinine, alkaline phosphatase, AST, ALT) for treatment groups were not significantly different from control values. Data are not shown for urine, hematology and clinical chemistry parameters.

Samples of all collected tissues for each rat in the subsets were not available for histopathological examination. Significant pathological changes were limited to squamous hyperplasia of the stomach and perianal dermatitis. The incidence and severity of these changes were found to be dose-dependent and statistically significant at 1500 mg/kg/day for perianal dermatitis and stomach hyperplasia (p<0.05, see Table 6). Only the incidence and severity of the squamous hyperplasia of the stomach were significantly increased after oral exposure to 750 mg/kg/day JP-8 (p<0.05).

One day after weaning, the dams were euthanized. A subset from each group was necropsied. Due to sacrifices falling on weekends and the deaths of three animals not related to JP-8 dose (two from the 325 and one from the 750 mg/kg/day groups), the number of female rats per subset was limited. Treatment subsets had 7 or 8 rats while the control subset had 10 rats available. Liver weights were significantly increased, as were liver to body weight and liver to brain weight ratios (p<0.01 in 1500 mg/kg/day group). Kidneys to brain ratios were also significantly increased (p<0.05).

Dose descriptor:

NOAEL

Remarks:

fertility

Effect level:

>= 1 500 mg/kg bw/day (actual dose received)

Sex:

female

Basis for effect level:

other: highest dose tested

Clinical signs:

no effects observed

Mortality / viability:

no mortality observed

Body weight and weight changes:

effects observed, treatment-related

Sexual maturation:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

not examined

Histopathological findings:

not examined

There was no difference in survival of pups between control and dose groups at PND 4, 14 and 21. Three pups, each from different litters in the 750 mg/kg/day dose group, did not survive between PND 21 and PND 90. Only one pup from the 1500 mg/kg/day group died during this time.

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

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

Sex:

male/female

Basis for effect level:

other: based on decreased pup weight

Reproductive effects observed:

not specified

Conclusions:

The NOAEL >=1500 mg/kg/day for female fertility.

Executive summary:

Female rats were dosed with neat JP-8 (0, 325, 750, 1500 mg/kg) daily by gavage for a total of 21 weeks (90-day plus mating with naive males, gestation and lactation) in an effort to assess general toxicity, fertility and reproductive endpoints. The NOAEL >=1500 mg/kg/day for female fertility. The NOAEL >=1500 mg/kg/day for female fertility. The NOAEL for the pup was 750 mg/kg/day based on a decrement in body weight which correlated with a decrease in maternal body weight at 1500 mg/kg/day.

Reference 2

Endpoint:

three-generation reproductive toxicity

Remarks:

based on test type (migrated information)

Type of information:

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

Adequacy of study:

key study

Study period:

1990

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Source of data is from peer reviewed literature. Acceptable well-documented study report which meets basic scientific principles. GLP.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 416 (Two-Generation Reproduction Toxicity Study)

Deviations:

yes

Remarks:

A three generational study examining reproductive toxicity.

Principles of method if other than guideline:

A three generational study examining reproductive toxicity.

GLP compliance:

yes

Limit test:

no

Species:

mouse

Strain:

CD-1

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Portage, MI
- Age at study initiation:(P) 6 x wks; (F1) 5-7 x wks; (F2) 22 days
- Housing: Inidividually housed in wire mesh cages
- Diet (e.g. ad libitum): Purina Certified Rodent Chow No. 5002 ad libitum except during exposure
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 2-3 weeks

ENVIRONMENTAL CONDITIONS
Animal husbandry followed standards by the US Department of Health, Education, and Welfare (1985)

Route of administration:

inhalation: vapour

Type of inhalation exposure (if applicable):

whole body

Vehicle:

unchanged (no vehicle)

Details on exposure:

- Exposure apparatus: 16 m glass and steel chambers.
- Method of holding animals in test chamber: cages
- Source and rate of air: Air was provided by a seperate HVAC system.
- Method of conditioning air: Air was filtered for particulates and temperature and humidity controlled.
- System of generating particulates/aerosols: Test atmosphere was generated by heating nitrogen to 200°C by passing it through a 1 l stainless steel cylinder with a 1500 W band heater. The nitrogen then passed through a glass column 7.6 cm diameter and 30 cm long packed with glass beads. Test material was delivered by a metering pump into Teflon tubing, to the bottom of the column. The liquid test substance vaporized as it went up the column with the nitrogen. The vapor then went into the test chambers where dilution with the chamber ventilation air produced the desired concentrations.
- Temperature, humidity, pressure in air chamber: Air flow rate, temperature and relative humidity were monitored every half-hour during exposure.

TEST ATMOSPHERE
- Brief description of analytical method used: Measurements made hourly using gas-phase IR.
- Samples taken from breathing zone: yes

Details on mating procedure:

- M/F ratio per cage: 1/1
- Length of cohabitation: 2 weeks
- Proof of pregnancy: sperm-positive vaginal smear referred to as day 0 of pregnancy

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Concentration of the test material in the test chambers was determined by GC analysis.

Frequency of treatment:

Both males and females were exposed 6 hours per day 5 days/ week for 10 weeks before mating. After mating, females were exposed 6 hours/day, 7 days/week from Gestation Day (GD) 0 to GD 20. Dams were then removed to nesting boxes to deliver. Dams were again exposed from postnatal day (LD) 5 until weaning on LD 21.

Details on study schedule:

- F1 parental animals not mated until 10 weeks after selected from the F1 litters.
- Selection of parents from F1 generation one week after weaning.
- Age at mating of the mated animals in the study: 15-17 weeks

Remarks:

Doses / Concentrations:
0, 100, 500, 1500 ppm
Basis:
nominal conc.
0, 500, 2500, 7500 mg/m3

Remarks:

Doses / Concentrations:
0, 103, 495, 1480 ppm
Basis:
analytical conc.
0, 515, 2475, 7400 mg/m3

No. of animals per sex per dose:

30, except for F2 generation in which 40 animals/sex/dose were selected. In the 1500 ppm F2 group all surviving animals were mated.

Control animals:

yes, concurrent no treatment

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Twice daily for viability and overt toxicity.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: weekly

BODY WEIGHT: Yes
- Time schedule for examinations: weekly until confirmation of mating; females weighed on GD 0, 7, 14, 21, and LD 0, 7, 14, 21

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: Yes, weekly except during mating, gestation, and lactation
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes / No / No data

Litter observations:

STANDARDISATION OF LITTERS
Litters were culled by random selection to 8 pups on LD4.


PARAMETERS EXAMINED
The following parameters were examined in F1/F2/F3 offspring:
Number of pups, stillbirths, live births, presence of gross anomalies was examined as soon as possible after delivery. Pups weighed individually on LD 0, 4, 7, 14. On LD21 all pups were counted, sexed, and weighed.


GROSS EXAMINATION OF DEAD PUPS:
Yes, culled pups and any pups that died were necropsied for gross abnormalities.

Postmortem examinations (parental animals):

SACRIFICE
- Male animals: All surviving animals were necropsied at end of mating period.
- Maternal animals: All surviving animals were necropsied following weaning.

HISTOPATHOLOGY
In control and high dose group the epididymis, lung, ovary, pituitary, prostate, seminal vesicle, testis, uterus, vagina, lymph nodes were examined. Masses and gross lesions were examined from all groups.

ORGAN WEIGHTS
Epididymis, lung, ovary, testis, prostate/seminal vesicle, uterus/vagina

Postmortem examinations (offspring):

SACRIFICE
- The F1/F2 offspring not selected as parental animals were sacrificed at 8 days of age. F3 offspring were sacrificed at LD21.

GROSS NECROPSY
Culled pups and any pups that died were necropsied for gross abnormalities.

HISTOPATHOLOGY / ORGAN WEIGTHS
F1/F2 offspring were examined the same as parental animals.

Statistics:

Fertility indices and female/male ratio was analysed using Chi-square test criterion with Yate's correction. Proportions of litters with malformations were compared using Fisher's exact probability test. Proportions of resorbed fetuses, implantation losses, and pup survival were compared using Mann-Whitney U test. Mean number of liveborn pups/litter and pup weight were compared using analysis of variance and appropriate t-tests. Other parameters were compared using analysis of variance, appropriate t-test, and Dunnett's multiple comparison tables.

Reproductive indices:

female mating index: number pregnant females/number of females mated
female conception index: number females delivering live litter/number pregnant females
female gestational index: number of females delivering live litter/number females delivering a litter
male fertility index: number of fertile males/number of males mated
Cohabitation time: average number male/female cohabitation days litter size at birth

Offspring viability indices:

Gestational survival index: number pups live born/number of pups born
Postnatal survival index (4-day): number pups alive at LD4/number of liveborn pups
Postnatal survival index (21-day): number pups alive at LD21/number of liveborn pups

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:

no effects observed

Reproductive function: oestrous cycle:

no effects observed

Reproductive function: sperm measures:

no effects observed

Reproductive performance:

no effects observed

F0
CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
All males survived with minimal signs of toxicity. 7 females in the 1500 ppm group died.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
Body weight gain of P males and females was significantly reduced in the 500 ppm and 1500 ppm groups. Food consumption was reduced in the 1500 ppm group in the first week.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
There were no significant differences in number of mated females, number of females delivering a litter, number of females delivering a live litter, or male fertility. The mean number of live births per litter were not significantly reduced. Length of time for mating was not significantly different from controls.

GROSS PATHOLOGY (PARENTAL ANIMALS)
There were no significant pathological findings in the reproductive organs in any group of animals in any generation in any dose group.

F1
CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
In the 1500 ppm group ataxia and reduced motor activity was observed. 6 females in this group also died.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
Mean body weights of 1500 ppm group, and males in the 500 ppm group were significantly less than control groups. Food consumption was normal.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
Length of time for mating was increased, but not significantly. Male fertility was reduced, however, as this effect was not seen in the first and third generations, this is not considered to be exposure related.

F2
CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
Most of the animals (36/40 males and 34/40 females) in the high exposure group (1500 ppm) died within the first week.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
Body weight gain of high exposure males and females were severely below control (up to 40%). Body weights in the 500 ppm and 100 ppm exposure groups were mildly depressed (10%). Food consumption was similar for all groups.

Dose descriptor:

NOAEC

Effect level:

>= 1 500 ppm (nominal)

Sex:

male/female

Basis for effect level:

other: 7500 mg/m3

Dose descriptor:

NOAEC

Effect level:

>= 1 500 ppm

Sex:

male/female

Basis for effect level:

other: 7500 mg/m3; If maternal exposure ceased before gestation day 20, there were no negative effects on the offspring.

Remarks on result:

other: Generation: offspring of all generations (migrated information)

Dose descriptor:

LOAEC

Effect level:

1 500 ppm

Sex:

male/female

Basis for effect level:

other: 7500 mg/m3; Reduced body weight in offspring was noted if maternal exposure was continued until delivery.

Remarks on result:

other: Generation: offspring of all generations (migrated information)

Clinical signs:

no effects observed

Mortality / viability:

no mortality observed

Body weight and weight changes:

effects observed, treatment-related

Sexual maturation:

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings:

no effects observed

F1
VIABILITY (OFFSPRING)
No significant differences through the lactation period.

BODY WEIGHT (OFFSPRING)
No significant difference in body weights or birth weight, except for pups from the 1500 ppm group at LD7 through weaning.

F2
VIABILITY (OFFSPRING)
Mean birth weight in the 1500 ppm group was reduced, but not significantly. The mean number of live offspring/litter, and number live-born/number delivered was significantly reduced in this group. This was most likely due to litters from dams which had not been confirmed to have mated. These unconfirmed dams were exposed until delivery, whereas the other dams were not exposed after GD20. The birth weights from these litters were not significantly reduced.

BODY WEIGHT (OFFSPRING)
Mean body weights in the 1500 ppm group after LD7 were significantly reduced.

F3
VIABILITY (OFFSPRING)
There were no apparent effects on mating, fertility, mean number of live births, and survival through lactation period.

BODY WEIGHT (OFFSPRING)
Mean weight of the offspring in the high exposure group was significantly reduced at birth. This may have been the result of the small sizes of the dams. If maternal exposure was ceased, the body weight gain became normal, but if reinitiated, the body weights were again reduced.

Reproductive effects observed:

not specified

Significant Effects of Exposure to High Flash Aromatic Naptha (SD)

Male Fertility Index	F0	F1	F2
0	86.7 (30)	89.7 (30)	93.3 (30)
100	96.7 (30)	86.7 (30)	83.3 (30)
500	83.3 (30)	93.3 (30)	80.0 (30)
1500	84.6 (26)	64.3 (28)	100 (4)
Litter Size at Birth	F0	F1	F2
0	12.1 ± 3.4	12.4 ± 2.0	12.6 ± 2.7
100	12.9 ± 1.5	11.1 ± 2.9	11.8 ± 2.3
500	12.2 ± 3.1	11.7 ± 3.0	11.4 ± 2.1
1500	11.3 ± 3.0	8.7 ± 4.3	12.2 ± 1.3
Gestational Survival Index	F1	F2	F3
0	95.9 (366)	97.4 (383)	97.7 (361)
100	97.9 (382)	95.4 (280)	98.2 (335)
500	94.9 (333)	91.6 (371)	98.5 (325)
1500	92.8 (279)	85.1 (215)	100 (73)

 

Gestational Survival Index Among Rats in the F2 Generation

Concentration	Total % (SD)	Prolonged Exposure (%) SD	Exposure Stopped on GD20 (%) SD
0	97.4 (383)	91.9 (74)	98.7 (309)
100	95.4 (280)	91.7 (96)	97.2 (184)
500	91.6 (371)	30.8 (13)	93.8 (358)
1500	85.1 (215)	63.0 (54)	92.5 (161)

 

Body Weights of Pups

Concentration (ppm)	F1	F2	F3
	Day 0
0	6.1± 0.5	6.0 ±0.5	6.0 ±0.5
100	6.2 ±0.5	6.1 ±0.5	6.0 ±0.4
500	6.5 ±0.6	6.0± 0.5	6.1 ±0.6
1500	6.1± 1.0	5.7 ±0.7	5.7 ±0.2
	Day 4
0	9.7 ±0.9	9.5 ±1.4	9.7 ±1.1
100	9.8 ±0.6	10.0± 1.2	10.0 ±0.7
500	10.1 ±1.0	9.9 ±1.0	9.8 ±1.0
1500	9.2 ±1.3	9.3 ±1.0	9.2 ±0.6
	Day 7
0	13.7 ±1.3	13.3 ±1.8	14.0±2.0
100	13.2 ±1.1	13.3 ±1.6	14.1 ±1.2
500	14.0 ±1.7	13.5 ±1.4	13.4 ±1.5
1500	12.0 ±1.8	11.7 ±1.3	12.0 ±1.0
	Day 14
0	24.9± 2.7	24.3 ±2.5	26.2± 4.0
100	23.2 ±1.8	23.5 ±2.8	25.6 ±1.9
500	23.9 ±2.4	23.7 ±2.7	23.2 ±2.7
1500	19.6 ±2.7	19.3 ±1.8	20.8 ±1.3
	Day 21 Male Body Weights
0	39.5 ±5.1	40.9 ±5.5	42.9 ±7.6
100	37.2 ±5.9	39.3 ±5.5	42.7 ±3.8
500	40.0 ±4.9	39.7 ±5.6	38.7± 5.1
1500	29.9 ±3.6	30.4 ±4.2	32.8 ±3.0
	Day 21 Female Body Weights
0	38.0 ±5.0	39.6 ±5.1	41.4 ±6.2
100	35.7 ±5.7	37.9 ±4.8	41.2 ±3.6
500	38.0 ±5.0	38.6 ±5.5	37.2 ±4.8
1500	29.4 ±4.3	29.1 ±4.2	31.8 ±3.6

Conclusions:

Concentrations of 1500 ppm (7500 mg/m3) caused reduced body weights in offspring of rats if maternal exposure was continued until delivery. If maternal exposure was ended on gestation day 20, no adverse effects were seen. The NOAEL was determined to be 1500 ppm (7500 mg/m3)

Executive summary:

Concentrations of 1500 ppm (7500 mg/m3) caused reduced body weights in offspring of rats if maternal exposure was continued until delivery. If maternal exposure was ended on gestation day 20, no adverse effects were seen. The results from the C9 aromatics represents the aromatic constituency of the C9 -C14 Aliphatics (2 -25% Aromatics). The NOAEL was determined to be 1500 ppm (7500 mg/m3).

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 500 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Quality of whole database:

Sufficient to meet data requirements. The study is Klimisch 2.

Effect on fertility: via inhalation route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEC

7 500 mg/m³

Study duration:

subchronic

Species:

mouse

Quality of whole database:

Sufficient to meet data requirements. The study is Klimisch 2.

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

The reproductive toxicity assessment of Alchisor TAL 111 is based on read-across data from C9-C14 aliphatics (2-25% aromatics).

 

In an OECD 415 oral fertility study onC9-C16 Aliphatics, 25% aromatics, the NOAEL was 1500 mg/kg for females and 3000 mg/kg for males. In a 3 generation inhalation fertility study on C9-C14 aliphatics (2-25% aromatics) the NOAEC was 7500 mg/m3.

 

The available reproductive toxicity studies gave rise to no significant reproductive toxicity by either the oral or inhalation routes.

 

 

 

 

 

Short description of key information:
The available reproductive toxicity studies gave rise to no significant reproductive toxicity by either the oral or inhalation routes.

Justification for selection of Effect on fertility via oral route:
The selected study is the longest duration study (one generation).

Justification for selection of Effect on fertility via inhalation route:
The selected study is a three generation study.

Effects on developmental toxicity

Description of key information

The available developmental toxicity study gave rise to no adverse maternal or fetal effects.

Link to relevant study records

Reference

Endpoint:

developmental toxicity

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

11 Sept. 1978 - 6 Oct. 1978

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Comparable to guideline study.

Qualifier:

according to guideline

Guideline:

other: Guidelines for Reproduction Studies for Safety and Evaluation of Drugs for Human Use, Segment II (Teratology Study)

GLP compliance:

no

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Breeding Laboratories, Inc.
- Age at study initiation: 7 weeks
- Fasting period before study: Animals were not given food during exposure.
- Housing: Individually, except during mating, in stainless steel cages, animals identified by ear tags
- Diet (e.g. ad libitum): Purina Lab Chow, ad libitum
- Water (e.g. ad libitum): Elizabethtown Water Company, ad libitum
- Acclimation period: Aug. 17, 1978-Sept. 4, 1978


ENVIRONMENTAL CONDITIONS
- Temperature (°C): monitored twice daily, room temperature
- Humidity (%): dry air
- Photoperiod (hrs dark / hrs light): 12 hrs light/12 hrs dark


IN-LIFE DATES: days 6-15 of gestation From: 11-27 Sept. 1978 To: 20 Sept. - 6 Oct. 1978

Route of administration:

inhalation: vapour

Type of inhalation exposure (if applicable):

whole body

Vehicle:

unchanged (no vehicle)

Details on exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: one cubic meter exposure chamber
- Temperature, humidity, pressure in air chamber: room temperature, dry air

Analytical verification of doses or concentrations:

no

Details on mating procedure:

- Impregnation procedure: cohoused
- If cohoused:
- M/F ratio per cage: 1/1
- Length of cohabitation: Overnight and removed in morning to check for pregnancy, this was repeated until females were pregnant
- Proof of pregnancy: vaginal plug and sperm in vaginal smear referred to as day 0 of pregnancy

Duration of treatment / exposure:

6 hrs/day

Frequency of treatment:

days 6-15 of gestation

Duration of test:

days 6-15 of gestation

Remarks:

Doses / Concentrations:
100 ppm
Basis:
nominal conc.
525 mg/m3

Remarks:

Doses / Concentrations:
300 ppm
Basis:
nominal conc.
1575 mg/m3

No. of animals per sex per dose:

20

Control animals:

yes, concurrent no treatment

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily
- Cage side observations: mortality, toxicological signs, pharmacological effects

BODY WEIGHT: Yes
- Time schedule for examinations: days 0, 6-15, 21 of gestation

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day #: 21
- Organs examined: uterus, ovaries

Ovaries and uterine content:

The ovaries and uterine content were examined after termination: Yes
Examinations included:
- Gravid uterus weight: No data
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
- Other: number of live fetuses, number of dead fetuses

Fetal examinations:

- External examinations: Yes: all per litter examined for sex, crown-rump distance, weighed, and malformations
- Soft tissue examinations: Yes: 2/3 per litter examined for gross dissection and examination of viscera, internal sex determination, ureter, kidneys, and heart
- Skeletal examinations: Yes: 2/3 per litter examined for skeletal malformations, and ossification
- Head examinations: Yes: 1/3 per litter examined for neural defects

Statistics:

Analysis was done using the chi-square method, or the F-test and Student's t-test. When the variance differed significantly, the Student's t-test was modified suing Chochran's approximation. The mean number of live fetuses, resorptions, implantations, and corpora lutea were analyzed using the one-tailed t-test.

Indices:

implantation efficiency,

Details on maternal toxic effects:

Maternal toxic effects:no effects

Details on maternal toxic effects:
There was no mortality in either of the dosage groups. Pregnancy rates were comparable between the exposure groups and negative controls. Weight gain was significantly higher in the exposure groups post-dosing. There were no significant clinical observations in either exposure group. The mean number of corpora lutea was significantly decreased in the 300 ppm group. Since ovulation occurred prior to exposure, this was not considered to be treatment related. The mean number of implantations was comparable between exposure groups and negative controls. The implantation efficiency values were actually significantly higher in exposure groups as compared to negative controls. The mean number of live fetuses, resorption sites, and number of dams with more than one resorption site were comparable between exposures and negative controls. The gross postmortem examination of dams showed no treatment related effects.

Dose descriptor:

NOAEC

Effect level:

>= 300 ppm

Basis for effect level:

other: developmental toxicity

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
The body weights of fetal males in the 100 ppm group were significantly higher than negative controls. There were some statistically significant differences in mean crown-rump distance between both dosage levels and negative controls, these differences were slight and the effect inconsistant between dosages and sex. These differences were therefore not considered to be indicative of a treatment related effect. Mean numbers of male and female fetuses, and sex ratios were similar between exposure groups and negative controls. Ossification variations were similar in exposure groups and negative controls, as was the incidence of litters with fetuses containing ossification variations. No malformations externally or in the soft tissues were noted in the fetuses except in the positive controls. Though skeletal defects were noted in the exposure group, the types of malformations are common in rat fetus and not considered to be treatment related.

Abnormalities:

not specified

Developmental effects observed:

not specified

Results - Dams

Endpoint	Negative Control	400 mg/kg ASA	100 ppm	300 ppm
Pregnancy Rate (%)	100.0	95.0	100.0	90.0
Mortality Rate (%)	0.0	10.0	0.0	0.0
Mean Body Weight Gain  (g) Dams - Days 15-21	84	32	108	103
Mean Corpora Lutea	15.2	14.5	15.5	13.8
Mean No. Implantations	13.0	13.2	13.8	13.2
Implantation Efficiency (%)	85.8	91.1	88.7	95.6
Mean No. Live Fetuses	12.5	7.4	12.9	12.6
Mean No. Dead Fetuses	0.0	0.0	0.0	0.0
Mean No. Resorptions	0.6	5.8	0.9	0.7
Dams with more than one Resorption (%)	10.0	58.8	25.0	11.1

Results – Fetuses

Endpoint	Negative Control	400 mg/kg ASA	100 ppm	300 ppm
Male Mean Fetal Weight (g)	5.57	3.88	5.82	5.62
Female Mean Fetal Weight (g)	5.29	3.62	5.44	5.33
Male Mean Crown-Rump Distance (cm)	4.3	3.7	4.4	4.2
Female Mean Crown-Rump Distance (cm)	4.2	3.6	4.2	4.1
Sex Ratio (%)	91.5	98.4	88.3	105.5
Ossification Variations (%)	70.7	100.0	79.4	79.3
Litters with Ossification Variations (%)	100.0	100.0	95.0	100.0
Soft Tissue Malformations (%)	2.4	26.8	1.1	3.9
Litters with Soft Tissue Malformations (%)	10.0	54.5	5.0	16.7
Gross Evisceration Malformations (%)	5.4	3.6	1.8	4.0
Litters with Gross Evisceration Malformations	25.0	8.3	10.0	16.7
Skeletal Malformations (%)	0.0	21.4	2.9	1.3
Litters with Skeletal Malformations	0.0	66.7	15.0	11.1

Conclusions:

The NOAEC for developmental toxicity in rats is >=300 ppm (1575 mg/m3) via inhalation. The test substance is also non-teratogenic.

Executive summary:

This study determined the developmental toxicity of MRD-78 -25 in rats exposed via inhalation. Groups of 20 pregnant female rats were exposed 6 hrs/day during days 6 -15 of gestation. Test concentrations of 100 or 300 ppm test substance. In addition to a negative control group, there was also a positive control group that was exposed to acetylsalicylic acid on days 6 -15 of gestation. Dams were observed for toxicological signs and pharmacological effects. On day 21 of gestation, the animals were sacrificed, and examined for corpora lutea and uterine implantation parameters. Fetuses were examined for fetal size, sex ratio, and external, soft-tissue, and skeletal malformations. No adverse effects due to exposure to the test substance were seen in either dams or fetuses. No treatment related malformation effects were noted in the fetuses. The developmental NOAEC for rats by inhalation is >=300 ppm. The test substance is also not teratogenic.

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

1 575 mg/m³

Study duration:

subacute

Species:

rat

Quality of whole database:

Sufficient to meet data requirements. The study is Klimisch 1.

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no study available

Additional information

The repeated dose toxicity assessment of Alchisor TAL 111 is based on read-across data from C9-C14 aliphatics (2-25% aromatics).

 

In a developmental toxicity study by the inhalation route the NOAEC was 1575 mg/m3. No adverse maternal or fetal effects were noted at any dose concentration.

 

 

Justification for selection of Effect on developmental toxicity: via inhalation route:
Only one study available.

Justification for classification or non-classification

Based on the available read-across data, the substance does not meet the criteria for classification.