Registration Dossier

Toxicological information

Toxicity to reproduction

Currently viewing:

Administrative data

Endpoint:
one-generation reproductive toxicity
Remarks:
based on generations indicated in Effect levels (migrated information)
Type of information:
migrated information: read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Study period:
2003 - 2008 (date final report signed by Study Director)
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP compliant (except for test substance identity, stability and lot number), guideline study , available as unpublished report, no restrictions, fully adequate for assessment

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2008
Report Date:
2008

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Deviations:
no
Qualifier:
according to
Guideline:
other: EPA OPPTS 870.3650 (Combined repeated exposure toxicity study with the Reproduction/Developmental toxicity screening test)
GLP compliance:
yes
Remarks:
The identity and stability was not in compliance with GLP regulations. The lot number was not available.
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): butane- Physical state: colourless gas- Analytical purity: 99.5% per supplier- Analytical purity: 99.23% before study, 99.98% after study- Lot/batch No.: not available- Expiration date of the lot/batch: not available- Stability under test conditions: confirmed- Storage condition of test material: Ambient - Other: An archival sample was stored.

Test animals

Species:
rat
Strain:
other: Sprague-Dawley CD
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS- Source: Charles River Laboratories, Raleigh, North Carolina 27610- Age at study initiation: Approximately 8 weeks- Weight at study initiation: males: 241-280 g (mean: 261 g); females 174-229 g (mean 200 g)- Fasting period before study: none- Housing: Individually in stainless steel suspended cages with wire mesh floors and fronts except during the mating period. From day 18 of gestation and during lactation, the each female was housed with her litter in plastic cages with bedding- Diet: Certified Rodent Diet # 5002 (PMI Nutrition Int., St. Louis, Missouri) ad libitum- Water: ad libitum- Acclimation period: Approximately 2 weeksENVIRONMENTAL CONDITIONS- Temperature 20.7-22.4°C- Humidity: 24.28-77.36%- Air changes (per hr): no data- Photoperiod: 12 hrs dark / 12 hrs lightIN-LIFE DATES: From: 29 September 2003 To: 26 April 2006

Administration / exposure

Route of administration:
inhalation
Type of inhalation exposure (if applicable):
whole body
Vehicle:
other: air
Details on exposure:
Not detailed in main report, require access to Appendix II, Volume 3GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION- Exposure apparatus: Whole-body inhalation chamber- Method of holding animals in test chamber: Individually in stainless steel suspended cages with wire mesh floors and fronts- Source and rate of air:- Method of conditioning air:- System of generating particulates/aerosols:- Temperature, humidity, pressure in air chamber:- Air flow rate:- Air change rate:- Method of particle size determination: yes- Treatment of exhaust air:TEST ATMOSPHERE- Brief description of analytical method used: Infrared spectrophotometer (IR) 4 times per chamber per day- Samples taken from breathing zone: yes
Details on mating procedure:
- M/F ratio per cage: 1:1 - Length of cohabitation: until evidence of mating was seen, or for two consecutive weeks - Proof of pregnancy: vaginal plug / sperm in vaginal smear referred to as day 0 of pregnancy- After successful mating each pregnant female was caged (how): individually in plastic "shoebox" cages with bedding- After the mating period was over, females without evidence of copulation were removed from the mating cages, housed individually and monitored for visible signs of pregnancy with corresponding bodyweight gain. - Any other deviations from standard protocol: no
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Exposure levels were determined using an infrared spectrophotometer 4 times/chamber/day. The test substance was evenly distributed within each chamber. The mean (± SD) analytical concentrations were 0 ± 0, 930.6 ± 28.1, 3022 ± 58 and 9157 ± 269 ppm.
Duration of treatment / exposure:
Males for 2 weeks prior to mating and for an additional 28 days (minimum) after mating.Females for 2 weeks prior to mating and gestation days 0-19.
Frequency of treatment:
6 hours/day, 7 days/week
Details on study schedule:
Females without evidence of mating that appeared to be pregnant were killed on an estimated gestation day 19.Females that littered and their offspring were killed on post partum day 4.
Doses / concentrationsopen allclose all
Remarks:
Doses / Concentrations:0, 900, 3000 and 9000 ppm Basis:other: target concentration
Remarks:
Doses / Concentrations:0.0 ± 0.0, 930.6 ± 28.1, 3022 ± 58, 9157 ± 269 ppm Basis:analytical conc.
No. of animals per sex per dose:
12/sex/group
Control animals:
yes, sham-exposed

Examinations

Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes- Time schedule: twice daily (mortality and clinical condition)DETAILED CLINICAL OBSERVATIONS: Yes- Time schedule: Male rats were examined prior to randomisation and once weekly throughout the study. Female rats were examined prior to randomisation and once weekly throughout the premating period and on gestation days 0, 7, 14, 20 and lactation days 0 (except if parturition was not completed on the same day), 1 and 4.BODY WEIGHT: Yes- Time schedule for examinations: Male rats were weighed at randomisation and then weekly throughout the study. Females were weighed at randomisation, on the first day of exposure and twice weekly until evidence of copulation was observed, on gestation days 0, 7, 14 and 20, and on lactation days 1 and 4. Females were not fasted prior to recording terminal bodyweights.FOOD CONSUMPTION: Yes- Time schedule for examinations: Once weekly throughout the premating period. For pregnant or confirmed mated females, food consumption was recorded on gestation days 0-7, 7-14, 14-20 and on lactation days 1-4.WATER CONSUMPTION: No
Estrous cyclicity (parental animals):
No
Sperm parameters (parental animals):
During the microscopic examination of the testes, special emphasis was placed on the stages of spermatogenesis and the histopathology of interstitial testicular cell structure.
Litter observations:
STANDARDISATION OF LITTERS- Performed on day 4 postpartum: noPARAMETERS EXAMINEDThe following parameters were examined in F1 offspring: Number of live and dead pups, pup abnormalities and sex of pups. Pups were observed twice daily (morning and afternoon).Each pup was examined on lactation day 0 and 4.Individual pup weights and sex were recorded on lactation days 1 and 4GROSS EXAMINATION OF DEAD PUPS:yes, for external and internal abnormalities; possible cause of death was not determined for pups born or found dead.
Postmortem examinations (parental animals):
SACRIFICE- Male animals: All surviving animals (after a minimum of 28 days post-mating).- Maternal animals: All surviving animals (28 days post-mating).GROSS EXAMINATION: Yes. Tissues examined: adrenal glands, bone (sternum/femur), bone marrow, brain (medulla/pons, cerebrum and cerebellum), epididymides, heart, kidneys, caecum, colon, rectum, larynx, liver, lungs (with mainstem bronchi), lymph nodes (mesenteric and mediastinal), mammary glands (with adjacent skin), nasopharynx, ovaries (with oviducts), prostate, seminal vesicles, duodenum, ileum, jejunum, spinal cord (cervical, thoracic and lumbar), spleen, stomach, testes, thymus, thyroid with parathyroids, tibial nerve, trachea, urinary bladder, uterus with vagina and all macroscopic lesions and tissue masses.ORGAN WEIGHTS: Yes. Adrenal glands, brain, epididymides, heart, kidneys, liver, lungs, ovaries, spleen, testes, thymus and uterus with vagina.HISTOPATHOLOGY: Yes. ORGAN WEIGHTS: Yes. Adrenal glands, brain, epididymides, heart, kidneys, liver, lungs, ovaries, spleen, testes, thymus and uterus with vagina.HISTOPATHOLOGY: Yes (male and female main study only). - tissues examined: adrenal glands, bone (sternum/femur), brain (cerebellum, cerebrum and cerebellum), epididymes, heart, kidneys, large intestine (caecum, colon and rectum), liver, lungs (with mainstream bronchi), lymph node (mesenteric), lymph node (mediastinal), mammary glands (with adjacent skin), ovaries (with oviducts), protate, seminal vesicles, smnall intestine (duodenum, ileum and jejunum), spinal cord (cervical, thoracic and lumbar), spleen, stomach, testes, thymus, thyroids with parathyroids, tibial nerve, trachea, urinary bladder, uterus with vagina, all macroscopic lesions and tissue masses.
Postmortem examinations (offspring):
Macroscopic postmortem examinations (external only) were performed on all surviving F1 pups on lactation day 4.
Statistics:
Group mean values of parameters for all the exposure groups were compared to the control group mean values at each time interval, using appropriate statistical methods.
Reproductive indices:
Male and female mating indices, pregnancy rates, male and female fertility indices, gestation indices and the incidence of dams with no viable pups, were analysed statistically.
Offspring viability indices:
Live birth index, litter survival and mean pup survival indices (days 0 and 4) were analysed statistically.

Results and discussion

Results: P0 (first parental animals)

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
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed

Reproductive function / performance (P0)

Reproductive performance:
no effects observed

Details on results (P0)

There were no effects on mating, fertility, gestational indices or reproductive performance in either male or female rats exposed to concentrations up to 9000 ppm butane for up to 6 weeks prior to, during, and after mating.

Effect levels (P0)

open allclose all
Dose descriptor:
NOAEC
Remarks:
Reproductive effects
Effect level:
9 000 ppm
Sex:
male/female
Dose descriptor:
NOAEC
Remarks:
Reproductive effects
Effect level:
21 394 mg/m³ air
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
Gross pathological findings:
no effects observed

Details on results (F1)

There were no effects on numbers of live or dead pups, pup abnormalities, sex or weight.

Effect levels (F1)

open allclose all
Dose descriptor:
NOAEC
Remarks:
Reproductive effects
Generation:
F1
Effect level:
9 000 ppm
Sex:
male/female
Dose descriptor:
NOAEC
Remarks:
Reproductive effects
Generation:
F1
Effect level:
21 394 mg/m³ air
Sex:
male/female

Overall reproductive toxicity

Reproductive effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
There were no effects on gestation duration, number of live and dead pups, pup abnormalities or pup sex and weights. The NOAEC for butane for reproductive toxicity was 9000 ppm. Equivalent to 21394 mg/m3
Executive summary:

The study assessed the potential toxicity, including neurotoxicity and reproductive performance in male and female rats following butane exposure at 900, 3000 and 9000 ppm. It also was designed to investigate effects inboth sexes on mating behavior and on gonadal function, as well as effects on conception, development, parturition and pup survival to lactation day 4.

Male and female rats were exposed for 6 hours/day, 7 days/week for 2 weeks prior to mating initiation. Main study females were evaluated for subchronic effects and were exposed once daily (6 hours/day), seven days/week for 4 weeks (28 days). A satellite group of females was evaluated for reproductive effects only - exposed once daily (6 hours/day), seven days/week for at least two weeks prior to mating initiation, then once daily during mating and gestation (days 0-19). For satellite female rats without evidence of mating that appeared to be pregnant, exposure was terminated on the estimated gestation day 19.Main study male rats were exposed during the mating and post-mating periods until euthanized for a minimum exposure of 28 days.

There was no effect on survival. There were no exposure-related clinical effects or effects on body weight, food consumption, FOB or motor activity parameters for either sex. Furthermore there were no exposure-related differences in haematology, clinical chemistry and no macroscopic or microscopic changes at post-mortem.

All mated female animals were found pregnant and delivered live pups. Mating indices for the butane male rats treated were comparable to control. Mating, fertility and gestation indices for the female rats were comparable to control. Most of the females in each group mated at the
first opportunity. There were also no treatment-related differences in the other reproductive parameters up to the time of parturition including the percent of females completing delivery and the duration of gestation. There were no treatment-related differences in all parturition parameters including the total number of pups delivered, the number of pups dying, the viability (4 day survival) index, the number of implantation sites and corpora lutea per dam, the pup sex ratio and the number of live pups/litter. The pups were unremarkable during the lactation period. There were no exposure-related differences in body weights or weight gains in the pups feeding from test substance exposed animals compared to the pups feeding from air control animals. Furthermore there were no macroscopic changes at post-mortem.

Exposure of male and female rats to target concentrations of 900, 3000 and 9000 ppm of butane resulted in no general systemic effects, no effects on fertility and reproductive performance and no effects on pup survival and development to postnatal Day4.               

A no observed adverse effect concentration (NOAEC) of 9000 ppm was determined for this study's systemic, reproductive, and neonatal endpoints. Equivalent to 21394 mg/m3 (MW 58.12g/mol).