Registration Dossier
Registration Dossier
Data platform availability banner - registered substances factsheets
Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.
The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.
Diss Factsheets
Use of this information is subject to copyright laws and may require the permission of the owner of the information, as described in the ECHA Legal Notice.
EC number: 231-639-5 | CAS number: 7664-93-9
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
![](https://echa.europa.eu./o/diss-blank-theme/images/factsheets/A-REACH/factsheet/print_toxicological-information.png)
Endpoint summary
Administrative data
Key value for chemical safety assessment
Effects on fertility
Description of key information
Reproduction/Developmental Toxicity Screening Test according to OECD Test guideline 421: NOEL 1000 mg/kg bw/day (Ceccatelli, R. 2010); key study, reliability 1.
One generation reproductive study in the mouse: NOAEL 5000 ppm (eq. 3044.15 mg/kg bw/day) for reproductive and general toxicity (Andres, C.J. and Cline, T.R, 1989); WoE, reliability 3.
One generation reproductive study in the pig:: NOAEL 3000 ppm for reproductive and general toxicity (Paterson, D.W. et al. 1979); WoE, reliability 3.
Link to relevant study records
- Endpoint:
- screening for reproductive / developmental toxicity
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- Experimental start and completion dates: 22 April 2010 to 30 August 2010
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
- Version / remarks:
- OECD guideline 421 (OECD guideline for testing of chemicals 421, “Reproduction/Developmental Toxicity Screening Test”, adapted by the Council on 27th July 1995.
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Limit test:
- no
- Justification for study design:
- To generate preliminary information using the OECD 421 "Reproduction/Developmental Toxicity Screening Test" concerning the effects of Sodium Sulphate on male and female reproductive performance such as gonadal function, mating behaviour, conception and parturition. Recognised by international guidelines as a recommended test system.
- Specific details on test material used for the study:
- SOURCE OF TEST MATERIAL
- Source (i.e. manufacturer or supplier) and lot/batch number of test material: Supplied by the Sponsor. Treuhandgemeinschaft Deutscher Chemiefasererzeuger GmbH (TDC)
- Purity: 99.5%
- Expiry Date (Retest Date): 30 Sep 2010
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room Temperature
- Stability and homogeneity of the test material in the vehicle/solvent under test conditions (e.g. in the exposure medium) and during storage: Application formulations were found to be homogeneously prepared with sufficient formulation stability under study storage conditions.
- Solubility and stability of the test material in the solvent/vehicle and the exposure medium: Soluble in highly purified water at formulated concentrations of 100, 300 & 1000 mg/kg bw/day. Stability of test formulations confirmed analytically.
- Reactivity of the test material with the incubation material used (e.g. plastic ware): None
TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing (e.g. warming, grinding): Formulated to concentration in purified water
OTHER SPECIFICS
- Other relevant information needed for characterising the tested material, e.g. if radiolabelled, adjustment of pH, osmolality and precipitate in the culture medium to which the test chemical is added: None - Species:
- rat
- Strain:
- Wistar
- Remarks:
- Rat, RccHan: WIST(SPF)
- Details on species / strain selection:
- Rat, RccHan: WIST(SPF)
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Harlan Laboratories, B.V. Kreuzelweg 53, 5961 NM Horst, Netherlands.
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 11 weeks
- Weight at study initiation: Males: 292 to 329 g, Females: 182 to 223 g
- Fasting period before study: No
- Housing: Individually in Makrolon type-3 cages with wire mesh tops and sterilized standard softwood bedding (‘Lignocel’ J.Rettenmaier & Söhne GmbH & CoKG, 73494 Rosenberg / Germany, imported by Provimi Kliba SA, 4303 Kaiseraugst, Switzerland). During the pre-pairing period, cages with males were interspersed amongst those holding females to promote the development of regular estrus cycles.
- Use of restrainers for preventing ingestion (if dermal): yes/no
- Diet (ad libitum): Pelleted standard Kliba Nafag 3433 rodent maintenance diet (Provimi Kliba SA, 4303 Kaiseraugst, Switzerland (Batch No's. 83/09 and 22/10.
- Water (ad libitum): Community tap-water from Füllinsdorf was available in water bottles.
- Acclimation period: 7 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3°C
- Humidity (%): 30 - 73.5%
- Air changes (per hr): Air-conditioned with 10 - 15 air changes/hr
- Photoperiod (hrs dark / hrs light): 12-hour fluorescent light / 12-hour dark cycle
IN-LIFE DATES: 29-Apr-2010 (test item administration) to 30-Aug-2010 (experimental completion) - Route of administration:
- oral: gavage
- Vehicle:
- water
- Remarks:
- Purified water
- Details on exposure:
- PREPARATION OF DOSING SOLUTIONS:
Dose formulations were prepared weekly. Sodium Sulphate was weighed into a glass beaker on a tared precision balance and approximately 80% of the vehicle was added (w/v). Using a magnetic stirrer, a homogeneous suspension was prepared. On obtaining an homogeneous mixture, the remaining vehicle was added. Separate formulations were prepared for each concentration. Dose formulations were stored at room temperature in brown glass beakers.
VEHICLE
- Purified water
- Concentration in vehicle: 10, 20 and 100 mg/kg dosed at a dose volume of 10 mL/kg bw.
- Amount of vehicle (if gavage): Dose volume of 10 mL/kg with a daily adjustment to actual body weight. - Details on mating procedure:
- - M/F ratio per cage: 1:1
- Length of cohabitation: During the pairing period, females were housed with sexually mature males (1:1) until evidence of copulation was observed.
- Proof of pregnancy: If the daily vaginal smear was sperm positive, or a copulation plug was observed. The day of mating was designated day 0 post coitum.
- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility.
- Further matings after two unsuccessful attempts: no
- After successful mating each pregnant female was caged (how): Individually
- Any other deviations from standard protocol: No - Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- Analysis of Dose Formulations:
On treatment day 1, samples from the control group as well as three samples (top, middle
and bottom) of about 2 g of each concentration were taken prior to dosing for analysis of
concentration and homogeneity. Samples of about 2 g of each concentration were taken from the
middle only to confirm stability (at 4 hrs and 7 days). During the second last week of the treatment, samples were taken from the middle to confirm concentration. The aliquots for analysis of dose formulations were frozen (-20 ± 5 °C) and delivered on dry ice to Dr. K. Morgenthal (Harlan Laboratories Ltd., Itingen, Switzerland) and stored there at -20 ± 5 °C until analysis. The samples were analyzed by HPLC coupled to a conductivity detector following an analytical procedure developed at Harlan Laboratories. The test item was used as the analytical standard. Duplicates were taken of all samples and were stored at Harlan Laboratories Ltd., Füllinsdorf, Switzerland.
The linearity of the analytical systems used for sample analyses was demonstrated with a good
relationship between peak areas measured and working standard concentrations. All calibration
points used met the acceptance limit of ±20% variation from the calibration curve derived by
linear regression analysis. The regression coefficients calculated were found to be better than
0.99.
The Sodium Sulphate peak was assigned in sample chromatograms by comparison to that of
working standards. In blank sample chromatograms no peak appeared at the retention time of
Sodium Sulphate and, therefore, the absence of the test item in the control samples was
confirmed.
The test item recoveries were found to be in the range of 86.7% to 97.4% with reference to the
nominal concentration. The maximum variation from the mean of homogeneity samples which were taken from the top, middle and bottom of the samples, ranged from 0.5% to 4.7%.
The stability of the application formulations was tested in samples taken four hours and seven
days after preparation and stored at room temperature. The variation values ranged from 0.2% to
3.2% from the time-zero value.
Results indicate the accurate use of the test item Sodium Sulphate and purified
water as vehicle during this study. Application formulations were found to be homogeneously
prepared and sufficient formulation stability under storage conditions was approved. - Duration of treatment / exposure:
- Duration of Treatment Period:
Males: 4 weeks
Females: ca. 7 weeks - Frequency of treatment:
- Once daily with treatment ending on day 3 post partum for females and on the day before sacrifice for males.
- Details on study schedule:
- Study Sequence Males Females
Acclimatization 7 days 7 days
First Test Item Administration Day 1 of pre-pairing Day 1 of pre-pairing
Pre-Pairing 14 days 14 days
Pairing 11 days maximum 11 days
Gestation Approximately 21 day -
Treatment Ends On day 3 post partum On day before sacrifice
Necropsy On day 4 post partum After a min. of 28 days treatment - Dose / conc.:
- 0 mg/kg bw/day (nominal)
- Remarks:
- Control group
- Dose / conc.:
- 100 mg/kg bw/day (nominal)
- Remarks:
- Low dose
- Dose / conc.:
- 300 mg/kg bw/day (nominal)
- Remarks:
- Middle dose
- Dose / conc.:
- 1 000 mg/kg bw/day (nominal)
- Remarks:
- High dose
- No. of animals per sex per dose:
- 40 males: 10 per group
40 females: 10 per group - Control animals:
- yes, concurrent vehicle
- Details on study design:
- - Dose selection rationale: The dose levels were selected based on a previous non-GLP dose range-finding toxicity study in Han Wistar rats, Harlan Laboratories Study C79092, using dose levels of 100, 300 and 1000 mg/kg/day, resulting in a NOEL of 300 mg/kg/day.
- Dose selection rationale:
- Rationale for animal assignment: By computer-generated random algorithm. - Positive control:
- Not applicable
- Parental animals: Observations and examinations:
- CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Viability / Mortality: Twice daily
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Clinical signs: Once daily, during acclimatization and up to day of necropsy.
BODY WEIGHT: Yes
- Time schedule for examinations: Recorded daily from treatment start to day of necropsy.
FOOD CONSUMPTION (oral gavage study, not a feeding (diet) study:
Males: Weekly during pre-pairing and after pairing periods.
Females: Pre-pairing period days 1 - 8 and 8 - 14, gestation period days 0 - 7, 7 - 14 and 14 - 21 post coitum and lactation period days 1 - 4 post partum.
No food consumption was recorded during the pairing period.
- Oestrous cyclicity (parental animals):
- During the pre-pairing period, cages with males were interspersed amongst those holding females to promote the development of regular estrus cycles.
During the pairing period, females were housed with sexually mature males (1:1) until evidence
of copulation was observed. The females were removed and housed individually if:
- the daily vaginal smear was sperm positive, or
- a copulation plug was observed.
The day of mating was designated day 0 post coitum. - Sperm parameters (parental animals):
- Parameters examined in [all/P/F1/F2] male parental generations:
[testis weight, epididymis weight, daily sperm production, sperm count in testes, sperm count in epididymides, enumeration of cauda epididymal sperm reserve, sperm motility, sperm morphology, other:] - Litter observations:
- STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: yes
All dams were allowed to give birth and rear their litters (F1 pups) up to day 4 post partum. Day 0 was designated as the day on which a female had delivered all her pups.
PARAMETERS EXAMINED
The following parameters were examined in F1 offspring: Litter size, live births, sex ratio, still births and any gross anomalies. Pups were weighed individually (without identification) on days 1 and 4 post partum.
GROSS EXAMINATION OF DEAD PUPS: No.
ASSESSMENT OF DEVELOPMENTAL NEUROTOXICITY: No.
ASSESSMENT OF DEVELOPMENTAL IMMUNOTOXICITY: No. - Postmortem examinations (parental animals):
- Slides of all organs and tissues collected at terminal sacrifice from the animals of the control and high-dose groups were examined by the study pathologist. The same applied to all occurring gross lesions. Special emphasis was made on the stages of spermatogenesis and histopathology of interstitial cell structure.
Histological examination of ovaries was carried out on any females (nos. 48, 49, 56 and 59) that did not give birth. Microscopic examination of the reproductive organs of all infertile males (nos.
8, 9, 16 and 19) was made. - Postmortem examinations (offspring):
- SACRIFICE
At the scheduled sacrifice, all animals were killed by an injection of sodium pentobarbital. Parental generation animals were exsanguinated.
GROSS NECROPSY
Pups were sacrificed on day 4 post partum. All pups were examined macroscopically for any structural changes at the scheduled necropsy.
HISTOPATHOLOGY / ORGAN WEIGTHS
Not applicable for the litter (only parental animals). - Statistics:
- The following statistical methods were used to analyze food consumption, body weights, reproduction data, macroscopical findings and organ weight:
• Means and standard deviations of various data were calculated.
• The Dunnett-test [see References (2)] (many to one t-test) based on a pooled variance estimate was applied if the variables could be assumed to follow a normal distribution for the comparison of the treated groups and the control groups for each sex.
• The Steel-test [see References (3)] (many-one rank test) was applied instead of the Dunnett-test when the data could not be assumed to follow a normal distribution.
• Fisher's exact-test [see References (4)] were applied if the variables could be dichotomized without loss of information. - Reproductive indices:
- Mating Performance and Fertility:
The median and mean precoital times were unaffected by test item treatment. Mean precoital times were 3.3, 4.3, 3.3 and 3.1 days in order of ascending dose level. The median precoital time was 3, 4, 3 and 3 days in order of ascending dose level.
Two females in groups 1 and two females in group 2 were not pregnant. Thus the fertility indices were 80.0%, (Group 1) 80.0% (Group 2), 100.0% (Group 3) and 100.0% (Group 4).
Fertility index = (Females achieving a pregnancy / Females paired) x 100 - Offspring viability indices:
- Postnatal Loss Days 0 - 4 Post Partum:
The total number of pup loss during the first four days was only one pup in the control group on
day 3. Therefore the viability indices were 98.9% in the control group and 100.0%, in groups 2,
3 and 4.
Viability index = (number of alive pups on day 4 p.p. / number of pups born alive) x 100. - Clinical signs:
- no effects observed
- Mortality:
- no mortality observed
- Body weight and weight changes:
- no effects observed
- Food consumption and compound intake (if feeding study):
- no effects observed
- Food efficiency:
- not examined
- Water consumption and compound intake (if drinking water study):
- not examined
- Ophthalmological findings:
- not examined
- Haematological findings:
- not examined
- Clinical biochemistry findings:
- not examined
- Endocrine findings:
- not examined
- Urinalysis findings:
- not examined
- Behaviour (functional findings):
- not examined
- Immunological findings:
- not examined
- Organ weight findings including organ / body weight ratios:
- no effects observed
- Histopathological findings: non-neoplastic:
- effects observed, non-treatment-related
- Description (incidence and severity):
- In group 4, three males were noted to have the pelvis of right kidney dilated.
In group 3, no abnormal findings were noted.
In group 2, one male was noted to have both testes and epididymides reduced in size.
In control group, one male was noted to have a pelvic dilation and enlargement of left kidney
and another male to have both testes and epididymides reduced in size.
Type and frequencies of these findings did not give an indication of a test item-related effect.
In testes of animal no. 9 in group 1 and no. 19 in group 2, marked to severe tubular atrophy was
found bilaterally and this finding was considered to be associated with their infertility. In the
above 2 animals, aspermia and cellular debris in epididymides which were considered to be
associated with the tubular atrophy of testes were also observed. No microscopic findings were
found in other reproductive organs of these animals, reproductive organs of other infertile males
(nos. 8 and 16) and ovaries of females (nos. 48, 49, 56 and 59) that did not give birth. - Histopathological findings: neoplastic:
- no effects observed
- Other effects:
- not specified
- Reproductive function: oestrous cycle:
- no effects observed
- Reproductive function: sperm measures:
- effects observed, non-treatment-related
- Description (incidence and severity):
- In the testes of animal no. 9 (Group 1) and no. 19 (Group 2), marked to severe tubular atrophy was found bilaterally and this finding was considered to be associated with their infertility. In both males, aspermia and cellular debris in epididymides which were considered to be associated with the tubular atrophy of testes were also observed. No microscopic findings were found in other reproductive organs of these animals, reproductive organs of other infertile males (nos. 8 and 16).
Treatment with the test item did not reaveal effects on the completeness of stages or cell
populations. There was no indication for maturation arrest or any other degenerative type. - Reproductive performance:
- no effects observed
- Description (incidence and severity):
- Mating performance, fertility index and conception rate were not affected by treatment with the
test item.
The mean number of corpora lutea, the mean number of implantations per dam, and the post implantation losses were unaffected by treatment with the test item. The mean duration of
gestation was also unaffected by treatment with the test item. - Key result
- Dose descriptor:
- NOEL
- Effect level:
- 1 000 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: An absence of any adverse effect, therefore the NOEL was established at 1000 mg/kg/day.
- Key result
- Critical effects observed:
- no
- Clinical signs:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Clinical signs at first litter check and during lactation were wound on ear or on nose or on mouth in one pup in group 1, in four pups in group 3 and one pup in group 4 and hind foot injured and toes missing in one pup in group 2. None of these findings gave an indication of a test item-related effect.
- Dermal irritation (if dermal study):
- not examined
- Mortality / viability:
- mortality observed, non-treatment-related
- Description (incidence and severity):
- The number of live pups at first litter check was unaffected by treatment with the test item. The number of pups found dead at first litter check was two pups each in the control group and in group 2.
The total number of pup loss during the first four days was only one pup in the control group on
day 3. Therefore the viability indices were 98.9% in the control group and 100.0%, in groups 2,
3 and 4. - Body weight and weight changes:
- no effects observed
- Food consumption and compound intake (if feeding study):
- no effects observed
- Food efficiency:
- not examined
- Water consumption and compound intake (if drinking water study):
- not examined
- Ophthalmological findings:
- not examined
- Haematological findings:
- not examined
- Clinical biochemistry findings:
- not examined
- Sexual maturation:
- not examined
- Anogenital distance (AGD):
- not examined
- Nipple retention in male pups:
- not examined
- Organ weight findings including organ / body weight ratios:
- no effects observed
- Gross pathological findings:
- no effects observed
- Histopathological findings:
- no effects observed
- Other effects:
- not specified
- Behaviour (functional findings):
- not examined
- Developmental immunotoxicity:
- not examined
- Key result
- Dose descriptor:
- NOEL
- Generation:
- F1
- Effect level:
- 1 000 mg/kg bw/day
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: No findings gave any indication of a test item-related effect.
- Remarks on result:
- not determinable due to absence of adverse toxic effects
- Key result
- Critical effects observed:
- no
- Key result
- Reproductive effects observed:
- no
- Treatment related:
- no
- Conclusions:
- As no effects were observed, the NOEL (No Observed Effect Level) for reproduction/developmental toxicity was considered to be 1000 mg/kg/day.
- Executive summary:
In this study, an OECD 421 "“Reproduction /Developmental Toxicity Screening Test” performed to GLP was conducted to generate preliminary information concerning the effects of the test item, Sodium Sulphate, on male and female reproductive performance such as gonadal function, mating behavior, conception and parturition.
Four groups (10 males & 10 females) were treated (by gavage) with Sodium Sulphate once daily. Group 1 was the purified water control, group 2, 3 and 4 were dosed at 100, 300 and 1000 mg/kg bw/day, respectively. All were dosed at a dose volume of 10 mL/kg bw. Males were treated over a 14-day pre-pairing period and during the pairing period up to one day before necropsy. Females were treated throughout pre-pairing, pairing, gestation and lactation period up to day 4 post partum.
In parental animals (P0), all animals survived until the scheduled necropsy and no clinical signs were observed. Mean food consumption was not affected by treatment in either sex. No test item-related effects on mean body weight and mean body weight gain were observeed in either sex. Mating performance, fertility index and conception rate were not affected by treatment. The mean number of corpora lutea, the mean number of implantations per dam, and the post-implantation losses were unaffected by test item treatment. The mean duration of gestation was unaffected by treatment. Mean weight of testes and epididymides were not affected by the treatment. All organs and tissues examined did not reveal any macroscopic or microscopic changes related to treatment.In F1 pups, the number of live pups at first litter check and the mean litter size was unaffected by test item treatment. Sex ratios at first litter check and on day 4 post partum were unaffected by treatment. Mean pup weights and weight gains were also not considered to be affected.
As no effects were observed, the NOEL (No Observed Effect Level) for reproduction/developmental toxicity was considered to be 1000 mg/kg/day.
- Endpoint:
- screening for reproductive / developmental toxicity
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Justification for type of information:
- No studies are proposed for scientific reasons or reasons of animal welfare given the corrosive nature of sulphuric acid. When added to water, sulphuric acid rapidly dissociates to the hydrogen and sulphate ions (pKa = 1.92), with the hydrogen ion responsible for localised irritation and corrosivity toxicity. As structurally sulphuric acid and sodium sulphate differ only by the cation this approach is considered acceptable. The data therefore adequately covers the reproductive toxicity endpoint following exposure to sodium sulphate.
- Reason / purpose for cross-reference:
- read-across source
- Sex:
- male/female
- Vehicle:
- water
- Remarks:
- Purified water
- Analytical verification of doses or concentrations:
- yes
- Parental animals: Observations and examinations:
- CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Viability / Mortality: Twice daily
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Clinical signs: Once daily, during acclimatization and up to day of necropsy.
BODY WEIGHT: Yes
- Time schedule for examinations: Recorded daily from treatment start to day of necropsy.
FOOD CONSUMPTION (oral gavage study, not a feeding (diet) study:
Males: Weekly during pre-pairing and after pairing periods.
Females: Pre-pairing period days 1 - 8 and 8 - 14, gestation period days 0 - 7, 7 - 14 and 14 - 21 post coitum and lactation period days 1 - 4 post partum.
No food consumption was recorded during the pairing period.
- Oestrous cyclicity (parental animals):
- During the pre-pairing period, cages with males were interspersed amongst those holding females to promote the development of regular estrus cycles.
During the pairing period, females were housed with sexually mature males (1:1) until evidence
of copulation was observed. The females were removed and housed individually if:
- the daily vaginal smear was sperm positive, or
- a copulation plug was observed.
The day of mating was designated day 0 post coitum. - Sperm parameters (parental animals):
- Parameters examined in [all/P/F1/F2] male parental generations:
[testis weight, epididymis weight, daily sperm production, sperm count in testes, sperm count in epididymides, enumeration of cauda epididymal sperm reserve, sperm motility, sperm morphology, other:] - Litter observations:
- STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: yes
All dams were allowed to give birth and rear their litters (F1 pups) up to day 4 post partum. Day 0 was designated as the day on which a female had delivered all her pups.
PARAMETERS EXAMINED
The following parameters were examined in F1 offspring: Litter size, live births, sex ratio, still births and any gross anomalies. Pups were weighed individually (without identification) on days 1 and 4 post partum.
GROSS EXAMINATION OF DEAD PUPS: No.
ASSESSMENT OF DEVELOPMENTAL NEUROTOXICITY: No.
ASSESSMENT OF DEVELOPMENTAL IMMUNOTOXICITY: No. - Postmortem examinations (parental animals):
- Slides of all organs and tissues collected at terminal sacrifice from the animals of the control and high-dose groups were examined by the study pathologist. The same applied to all occurring gross lesions. Special emphasis was made on the stages of spermatogenesis and histopathology of interstitial cell structure.
Histological examination of ovaries was carried out on any females (nos. 48, 49, 56 and 59) that did not give birth. Microscopic examination of the reproductive organs of all infertile males (nos.
8, 9, 16 and 19) was made. - Postmortem examinations (offspring):
- SACRIFICE
At the scheduled sacrifice, all animals were killed by an injection of sodium pentobarbital. Parental generation animals were exsanguinated.
GROSS NECROPSY
Pups were sacrificed on day 4 post partum. All pups were examined macroscopically for any structural changes at the scheduled necropsy.
HISTOPATHOLOGY / ORGAN WEIGTHS
Not applicable for the litter (only parental animals). - Statistics:
- The following statistical methods were used to analyze food consumption, body weights, reproduction data, macroscopical findings and organ weight:
• Means and standard deviations of various data were calculated.
• The Dunnett-test [see References (2)] (many to one t-test) based on a pooled variance estimate was applied if the variables could be assumed to follow a normal distribution for the comparison of the treated groups and the control groups for each sex.
• The Steel-test [see References (3)] (many-one rank test) was applied instead of the Dunnett-test when the data could not be assumed to follow a normal distribution.
• Fisher's exact-test [see References (4)] were applied if the variables could be dichotomized without loss of information. - Reproductive indices:
- Mating Performance and Fertility:
The median and mean precoital times were unaffected by test item treatment. Mean precoital times were 3.3, 4.3, 3.3 and 3.1 days in order of ascending dose level. The median precoital time was 3, 4, 3 and 3 days in order of ascending dose level.
Two females in groups 1 and two females in group 2 were not pregnant. Thus the fertility indices were 80.0%, (Group 1) 80.0% (Group 2), 100.0% (Group 3) and 100.0% (Group 4).
Fertility index = (Females achieving a pregnancy / Females paired) x 100 - Offspring viability indices:
- Postnatal Loss Days 0 - 4 Post Partum:
The total number of pup loss during the first four days was only one pup in the control group on
day 3. Therefore the viability indices were 98.9% in the control group and 100.0%, in groups 2,
3 and 4.
Viability index = (number of alive pups on day 4 p.p. / number of pups born alive) x 100. - Clinical signs:
- no effects observed
- Mortality:
- no mortality observed
- Body weight and weight changes:
- no effects observed
- Organ weight findings including organ / body weight ratios:
- no effects observed
- Reproductive function: sperm measures:
- effects observed, non-treatment-related
- Description (incidence and severity):
- In the testes of animal no. 9 (Group 1) and no. 19 (Group 2), marked to severe tubular atrophy was found bilaterally and this finding was considered to be associated with their infertility. In both males, aspermia and cellular debris in epididymides which were considered to be associated with the tubular atrophy of testes were also observed. No microscopic findings were found in other reproductive organs of these animals, reproductive organs of other infertile males (nos. 8 and 16).
Treatment with the test item did not reaveal effects on the completeness of stages or cell
populations. There was no indication for maturation arrest or any other degenerative type. - Reproductive performance:
- no effects observed
- Description (incidence and severity):
- Mating performance, fertility index and conception rate were not affected by treatment with the
test item.
The mean number of corpora lutea, the mean number of implantations per dam, and the post implantation losses were unaffected by treatment with the test item. The mean duration of
gestation was also unaffected by treatment with the test item. - Key result
- Dose descriptor:
- NOEL
- Effect level:
- 1 000 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: An absence of any adverse effect, therefore the NOEL was established at 1000 mg/kg/day.
- Clinical signs:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Clinical signs at first litter check and during lactation were wound on ear or on nose or on mouth in one pup in group 1, in four pups in group 3 and one pup in group 4 and hind foot injured and toes missing in one pup in group 2. None of these findings gave an indication of a test item-related effect.
- Mortality / viability:
- mortality observed, non-treatment-related
- Description (incidence and severity):
- The number of live pups at first litter check was unaffected by treatment with the test item. The number of pups found dead at first litter check was two pups each in the control group and in group 2.
The total number of pup loss during the first four days was only one pup in the control group on
day 3. Therefore the viability indices were 98.9% in the control group and 100.0%, in groups 2,
3 and 4. - Key result
- Dose descriptor:
- other:
- Generation:
- F1
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: No findings gave any indication of a test item-related effect.
- Remarks on result:
- not determinable due to absence of adverse toxic effects
- Reproductive effects observed:
- no
- Treatment related:
- no
- Conclusions:
- As no effects were observed, the NOEL (No Observed Effect Level) for reproduction/developmental toxicity was considered to be 1000 mg/kg/day.
- Executive summary:
In this study, an OECD 421 "“Reproduction /Developmental Toxicity Screening Test” performed to GLP was conducted to generate preliminary information concerning the effects of the test item, Sodium Sulphate, on male and female reproductive performance such as gonadal function, mating behavior, conception and parturition.
Four groups (10 males & 10 females) were treated (by gavage) with Sodium Sulphate once daily. Group 1 was the purified water control, group 2, 3 and 4 were dosed at 100, 300 and 1000 mg/kg bw/day, respectively. All were dosed at a dose volume of 10 mL/kg bw. Males were treated over a 14-day pre-pairing period and during the pairing period up to one day before necropsy. Females were treated throughout pre-pairing, pairing, gestation and lactation period up to day 4 post partum.
In parental animals (P0), all animals survived until the scheduled necropsy and no clinical signs were observed. Mean food consumption was not affected by treatment in either sex. No test item-related effects on mean body weight and mean body weight gain were observeed in either sex. Mating performance, fertility index and conception rate were not affected by treatment. The mean number of corpora lutea, the mean number of implantations per dam, and the post-implantation losses were unaffected by test item treatment. The mean duration of gestation was unaffected by treatment. Mean weight of testes and epididymides were not affected by the treatment. All organs and tissues examined did not reveal any macroscopic or microscopic changes related to treatment.In F1 pups, the number of live pups at first litter check and the mean litter size was unaffected by test item treatment. Sex ratios at first litter check and on day 4 post partum were unaffected by treatment. Mean pup weights and weight gains were also not considered to be affected.
As no effects were observed, the NOEL (No Observed Effect Level) for reproduction/developmental toxicity was considered to be 1000 mg/kg/day.
- Endpoint:
- one-generation reproductive toxicity
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- weight of evidence
- Study period:
- Published 1979
- Reliability:
- 3 (not reliable)
- Rationale for reliability incl. deficiencies:
- significant methodological deficiencies
- Justification for type of information:
- No studies are proposed for scientific reasons or reasons of animal welfare given the corrosive nature of sulphuric acid. When added to water, sulphuric acid rapidly dissociates to the hydrogen and sulphate ions (pKa = 1.92), with the hydrogen ion responsible for localised irritation and corrosivity toxicity. As structurally sulphuric acid and sodium sulphate differ only by the cation this approach is considered acceptable. The data therefore adequately covers the reproductive toxicity endpoint following high exposure to sulphate in drinking water.
- Reason / purpose for cross-reference:
- read-across source
- Clinical signs:
- no effects observed
- Mortality:
- mortality observed, non-treatment-related
- Body weight and weight changes:
- no effects observed
- Water consumption and compound intake (if drinking water study):
- effects observed, treatment-related
- Description (incidence and severity):
- Water consumption did not differ during gestation but increased during lactation as salt level increased
- Reproductive function: oestrous cycle:
- not examined
- Reproductive performance:
- no effects observed
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- 3 320 ppm
- Based on:
- test mat.
- Sex:
- female
- Remarks on result:
- not determinable because of methodological limitations
- Clinical signs:
- no effects observed
- Mortality / viability:
- mortality observed, non-treatment-related
- Body weight and weight changes:
- no effects observed
- Food consumption and compound intake (if feeding study):
- no effects observed
- Food efficiency:
- not examined
- Ophthalmological findings:
- not examined
- Haematological findings:
- no effects observed
- Clinical biochemistry findings:
- no effects observed
- Urinalysis findings:
- not examined
- Sexual maturation:
- not examined
- Anogenital distance (AGD):
- not examined
- Organ weight findings including organ / body weight ratios:
- not examined
- Gross pathological findings:
- not examined
- Histopathological findings:
- not examined
- Behaviour (functional findings):
- not examined
- Key result
- Dose descriptor:
- NOAEL
- Generation:
- F1
- Effect level:
- 3 000 ppm
- Based on:
- test mat.
- Sex:
- male/female
- Remarks on result:
- not determinable because of methodological limitations
- Key result
- Critical effects observed:
- not specified
- Key result
- Reproductive effects observed:
- not specified
- Conclusions:
- Study results suggest that sulfates up to and including 3320 ppm in water have no significant
effect on reproduction either in the gilt or sow. Furthermore, no significant treatment differences occurred in average daily gain or feed and gain ratio in offspring after weaning administered high sulfate levels of 3000 ppm with sulfate (from sodium sulfate) or 3000 ppm sulphate (equally from sodium and magnesium sulfate). - Executive summary:
In this study, thirty-one sows and 27 gilts were each assigned to three treatments groups to study the effect of water quality during gestation and lactation. Sodium sulfate was added to the water to give sulfate at 320 ppm, 1820 ppm and 3320 ppm. Water was offered ad libitum from about 30 days postbreeding through 28 days lactation. It was concluded that sulfate content of water consumed during
gestation had no significant effect on gestation gain, number of pigs per litter at birth (total and live) or average pig and litter birth weights. Sulfate in water during lactation also had no sigificant effect on lactation gain, number of pigs at 28 days and average pig and litter weights at 28 days. The ingestion of sulfates in water upto 3320 ppm had no effect of reproductive performance. When water quality was evaluated using the offspring after weaning, no significant differences occurred at 28 days in average daily gain or feed to gain ratio among weaned pigs that consumed 3000 ppm sulfate (from sodium sulfate) or 3000 ppm sulphate (equally from sodium and magnesium sulfate).- Endpoint:
- one-generation reproductive toxicity
- Type of information:
- other: Journal publication
- Adequacy of study:
- weight of evidence
- Study period:
- Published 1979
- Reliability:
- 3 (not reliable)
- Rationale for reliability incl. deficiencies:
- significant methodological deficiencies
- Qualifier:
- no guideline followed
- Principles of method if other than guideline:
- Thirty-one sows and 27 gilts were each assigned to three treatments to study the effect of sulfate water quality during gestation and lactation. Sodium sulfate was added to local water to give sulfate at: 320 ppm, 1820 ppm and 3320 ppm. Water was offered ad libitum from about 30 days post breeding through to day 28 of lactation. Fifty-four weaned pigs (representing the three treatment groups) were given water with 0 ppm, 3000 ppm (sulfate from sodium sulfate) or 3000 ppm (1:1 sulfate from magnesium and sodium sulfate) for a 28-day period.
Parameters evaluated were gestation or lactation gains, number or weight of pigs at birth or at weaning, water consumption, fecal consistency, daily gain
or feed to gain ratio. - GLP compliance:
- no
- Limit test:
- no
- Justification for study design:
- - Premating exposure duration for parental (P0) animals: Water was offered (ad libitum) from approximately 30 days post breeding through to lactation day 28.
- Basis for dose level selection: Three sulfate concentrations, 320 ppm, 1820 ppm and 3320 ppm
- Inclusion/exclusion of extension of Cohort 1B. No, non-standard study (1989) which does not meet current guideline requirements.
- Termination time: Not stated
- Route of administration: Ad libitum via drinking water
- Other considerations:
Low number of pigs per group which does not reflect current guidance requirements.
Limited reproductive parameters recorded which does not reflect current guidance requirements.
The pig is considered an applicable experimental model.
In addition, an evaluation of the effect of water quality on offspring after weaning was examined. 54 four week-old pigs averaging 7.5 to 8.0 kg were assigned into nine groups. Each group consisted of two pigs from each of the three sow treatments. These groups were randomly allotted to three replications of three treatments:
1: Control water
2: 3000 ppm of added sulfate from sodium sulfate
3: 3000 ppm of added sulfate supplied equally from magnesium and sodium sulfate.
Each 2.4 x 3 m pen contained six pigs. All pigs were offered water and an 18% protein, fortified corn-soybean meal diet ad libitum for the 28-day trial. - Specific details on test material used for the study:
- Sulfate in the form of analytical grade sodium sulfate was added as appropriate to achieve the desired concentrations.
- Species:
- pig
- Strain:
- not specified
- Details on species / strain selection:
- 31 sows and 27 gilts of Hampshire, Yorkshire and Duroc breeds.
- Sex:
- female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- The reproductive trial involved 31 sows and 27 gilts, grouped separately on the basis of ancestry and weight.
- Source: Not stated
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: Not stated
- Weight at study initiation: Not stated
- Fasting period before study: Not stated
- Housing: Housed in unsulfated , wooden, colony type houses located in dry lots.
- Diet (ad libitum): No, each animal was restricted to 1.8 kg of feed/per day via individual feeding stalls. Fortified corn-soyabean meal diets with 10% alfalfa meals (gestation) and 10% beet pulp (lactation) included 0.5% trace mineralized salt. Crude protein content was 12.65% and 15.70% respectively.
- Water (ad libitum): yes, available from 227 litre circular tank waterers.
Self-feeders containing the lactation diet and the 227 waterers were located distant to their housing. Animals were allowed access for 2 hours in the morning and 1.5 hours in the evening.
- Acclimation period: Not stated
ENVIRONMENTAL CONDITIONS
- Temperature (°C), Humidity (%), Photoperiod (hrs dark / hrs light): Not stated
IN-LIFE DATES: From: To: Not stated - Route of administration:
- oral: drinking water
- Vehicle:
- water
- Remarks:
- The local water supply was used both for the control and for making the experimental waters.
- Details on exposure:
- In sows and gilts water (containing sulfates at 320, 1820 and 3320 ppm) was offered (ad libitum) from approximately 30 days post breeding through to lactation day 28.
To examine the effects of high sulfate levels on offspring after weaning, 54 weaned pigs representing the above three sow treatments were given water with 0 sulfates (Control), or 3,000 ppm sulfate (from sodium sulfate ) or 3,000 ppm sulfate (from magnesium and sodium sulfate, 1:1 ratio) for a 28-day period. - Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- Sulfate content was determined weekly by turbidimetric method. Averages with their standard deviations for the entire experimental period were: Control, 320 (±24) ppm; low sulfate: 1790 (±35) ppm and high sulfate, 3298 (±139) ppm.
- Duration of treatment / exposure:
- For 30 days post-breeding sows and gilts were randomly assigned to three groups, a control group (320 ppm sulfate), a low sulfate group (1820 ppm sulfate) and high sulfate group (3320 ppm sulfate). Water was available ad libitum from 30-days post-breeding through to 28-days post-lactation. During gestation, feed was restricted to a daily amount of 1.8 kg/per head with sows allowed access for 2 hrs (am) and 1.5 hrs (pm). After parturition, offspring at 4-weeks were placed into three treatment groups to evaluate sulfate administration to offspring. The groups were, 1: (Control water), 2: (3000 ppm sulfate from sodium sulfate) and 3: (3000 ppm sulfate, 1:1 magnesium and sodium sulfate) for an exposure period of 28-days.
- Frequency of treatment:
- Exposure period was daily (ad libitum in water) from 30-days post breeding to 28-days post-lactation.
- Details on study schedule:
- The trial involved 31 sows and 27 gilts (grouped separately) which were randomly assigned to three treatment groups ca. 30 days post breeding. Local water was used as the control and for treatment formulations. The three three treatment groups were 320 ppm (control), 1820 ppm (low sulphate) and 3320 ppm (high sulfate). Analysis used a turbidimetric method. During gestation, feed was restricted to 1.8 kg/animal daily with water available ad libitum. Sows were allowed access to feed and water each morning and evening for 2 hrs and 1.5 hrs respectively. Saline water was available to pigs from a creep area after 10 days of age.
At parturition, the number of live and stillborn, litter weight and average pig weight were recorded. Litter weight at 14 days, number of pigs at 28 days, litter weight and average pig weight at 28 days were recorded.
To determine the effect of water quality on offspring after weaning, 54 four-week-old pigs were allotted to three treatment groups: a control group (water), a sulfate group of 3000 ppm of added sulfate from sodium sulfate and a high sulfate group of 3000 ppm of added sulfate 1:1 from magnesium and sodium sulfate. All pigs were offered water and diet ad libitum for the 28-day trial. - Dose / conc.:
- 320 ppm (nominal)
- Remarks:
- Control
- Dose / conc.:
- 1 820 ppm (nominal)
- Remarks:
- Sulfate (Low) treatment group
- Dose / conc.:
- 3 320 ppm (nominal)
- Remarks:
- Sulfate (High) treatment group
- Control animals:
- yes, concurrent vehicle
- Details on study design:
- - Dose selection rationale: Three sulphate groups,
1: Control: Local water - Sulfate 320ppm
2: Sulfate (Low) group - 1820 ppm
3: Sulfate (High) group - 3320 ppm
- Rationale for animal assignment (if not random): Random
- Fasting period before blood sampling for clinical biochemistry: Not stated - Parental animals: Observations and examinations:
- CAGE SIDE OBSERVATIONS: No
DETAILED CLINICAL OBSERVATIONS: No
BODY WEIGHT: Yes
WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes - Litter observations:
- At parturition, the number of live and stillborn pigs as well as litter weight and average pig weight were obtained.
Litter weight at 14 days, number of pigs at 28 days, litter weight and average pig weight at 28 days were recorded. - Statistics:
- Data were analyzed by least squares analysis of variance.
- Reproductive indices:
- Sulfates up to and including 3320 ppm in water have no significant effect on reproduction performance in the gilt or sow.
- Offspring viability indices:
- Sulfate content of water consumed during gestation had no significant effect on gestation gain, number of pigs per litter at birth (total and live) or average pig and litter birth weights.
- Clinical signs:
- no effects observed
- Mortality:
- mortality observed, non-treatment-related
- Body weight and weight changes:
- no effects observed
- Water consumption and compound intake (if drinking water study):
- effects observed, treatment-related
- Description (incidence and severity):
- Water consumption did not differ during gestation but increased during lactation as salt level increased
- Reproductive function: oestrous cycle:
- not examined
- Reproductive performance:
- no effects observed
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- 3 320 ppm
- Based on:
- test mat.
- Sex:
- female
- Remarks on result:
- not determinable because of methodological limitations
- Clinical signs:
- no effects observed
- Mortality / viability:
- mortality observed, non-treatment-related
- Body weight and weight changes:
- no effects observed
- Food consumption and compound intake (if feeding study):
- no effects observed
- Food efficiency:
- not examined
- Ophthalmological findings:
- not examined
- Haematological findings:
- no effects observed
- Clinical biochemistry findings:
- no effects observed
- Urinalysis findings:
- not examined
- Sexual maturation:
- not examined
- Anogenital distance (AGD):
- not examined
- Organ weight findings including organ / body weight ratios:
- not examined
- Gross pathological findings:
- not examined
- Histopathological findings:
- not examined
- Behaviour (functional findings):
- not examined
- Key result
- Dose descriptor:
- NOAEL
- Generation:
- F1
- Effect level:
- 3 000 ppm
- Based on:
- test mat.
- Sex:
- male/female
- Remarks on result:
- not determinable because of methodological limitations
- Key result
- Critical effects observed:
- not specified
- Key result
- Reproductive effects observed:
- not specified
- Conclusions:
- Study results suggest that sulfates up to and including 3320 ppm in water have no significant
effect on reproduction either in the gilt or sow. Furthermore, no significant treatment differences occurred in average daily gain or feed and gain ratio in offspring after weaning administered high sulfate levels of 3000 ppm with sulfate (from sodium sulfate) or 3000 ppm sulphate (equally from sodium and magnesium sulfate). - Executive summary:
In this study, thirty-one sows and 27 gilts were each assigned to three treatments groups to study the effect of water quality during gestation and lactation. Sodium sulfate was added to the water to give sulfate at 320 ppm, 1820 ppm and 3320 ppm. Water was offered ad libitum from about 30 days postbreeding through 28 days lactation. It was concluded that sulfate content of water consumed during
gestation had no significant effect on gestation gain, number of pigs per litter at birth (total and live) or average pig and litter birth weights. Sulfate in water during lactation also had no sigificant effect on lactation gain, number of pigs at 28 days and average pig and litter weights at 28 days. The ingestion of sulfates in water upto 3320 ppm had no effect of reproductive performance. When water quality was evaluated using the offspring after weaning, no significant differences occurred at 28 days in average daily gain or feed to gain ratio among weaned pigs that consumed 3000 ppm sulfate (from sodium sulfate) or 3000 ppm sulphate (equally from sodium and magnesium sulfate).- Endpoint:
- one-generation reproductive toxicity
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- weight of evidence
- Justification for type of information:
- No studies are proposed for scientific reasons or reasons of animal welfare given the corrosive nature of sulphuric acid. When added to water, sulphuric acid rapidly dissociates to the hydrogen and sulphate ions (pKa = 1.92), with the hydrogen ion responsible for localised irritation and corrosivity toxicity. As structurally sulphuric acid and sodium sulphate differ only by the cation this approach is considered acceptable. The data therefore adequately covers the reproductive toxicity endpoint following high exposure to sulphate in drinking water.
- Reason / purpose for cross-reference:
- read-across source
- Specific details on test material used for the study:
- Available study data for sodium sulphate is being used for read-across to the target substance, sulphuric acid.
- Clinical signs:
- no effects observed
- Mortality:
- no mortality observed
- Body weight and weight changes:
- effects observed, non-treatment-related
- Reproductive function: oestrous cycle:
- no effects observed
- Reproductive function: sperm measures:
- not examined
- Reproductive performance:
- effects observed, non-treatment-related
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- 5 000 ppm
- Based on:
- test mat.
- Sex:
- female
- Remarks on result:
- not determinable because of methodological limitations
- Key result
- Critical effects observed:
- not specified
- Clinical signs:
- no effects observed
- Mortality / viability:
- no mortality observed
- Body weight and weight changes:
- effects observed, non-treatment-related
- Food consumption and compound intake (if feeding study):
- not examined
- Food efficiency:
- not examined
- Water consumption and compound intake (if drinking water study):
- effects observed, non-treatment-related
- Ophthalmological findings:
- not examined
- Haematological findings:
- not examined
- Clinical biochemistry findings:
- no effects observed
- Urinalysis findings:
- not examined
- Sexual maturation:
- not examined
- Anogenital distance (AGD):
- not examined
- Nipple retention in male pups:
- not examined
- Organ weight findings including organ / body weight ratios:
- not examined
- Gross pathological findings:
- not examined
- Histopathological findings:
- not examined
- Other effects:
- not specified
- Behaviour (functional findings):
- not examined
- Developmental immunotoxicity:
- not examined
- Key result
- Dose descriptor:
- NOAEL
- Generation:
- F1
- Effect level:
- 5 000 ppm
- Based on:
- test mat.
- Sex:
- male/female
- Remarks on result:
- not determinable because of methodological limitations
- Key result
- Critical effects observed:
- not specified
- Key result
- Reproductive effects observed:
- not specified
- Conclusions:
- No effect on reproductive performance was observed in female mice following the ad libitum ingestion of sulfate (from sodium sulfate) in drinking water at up to 5000 ppm during gestation or lactation over two parities.
- Executive summary:
In this study, six treatment groups of 10 female mice were administered water (ad libitum) as either: (0)Control (deionized water), (0)Sodium control (in deionised water) or 625 ppm, 1250 ppm, 2500 ppm or 5000 ppm sulfate. Concentrations were adapted by the addition of sodium bicarbonate content. Premating exposure of parental females to treated water was for 1 week prior to the addition of males used for mating, on confirmation of mating males were removed. All parental females were dosed throughout until study termination. Each offspring was treated from day 21 post-partum until study termination. No effect on reproductive performance were observed in female mice during gestation or lactation over two parities. Sulfate (from sodium sulfate) administered at concentrations up to 5000 ppm (ad libitum in drinking water) caused no effect on litter size, litter weaning weight, or gestational or lactational weight gain of dams over two parities. Gestational and lactational gain of dams
was not affected by level of sulfate consumed. Minor decreases in water consumption with increasing treatment (compared to the sodium control group) were noted during both parities. Also, weight gain in dams was greater during the first parity and a quadratic response to sulfates in litter weaning weights was observed in the second parity. There were no adverse effects on reproductive performance.- Endpoint:
- one-generation reproductive toxicity
- Type of information:
- other: Journal publication
- Adequacy of study:
- weight of evidence
- Study period:
- Published 1989
- Reliability:
- 3 (not reliable)
- Rationale for reliability incl. deficiencies:
- significant methodological deficiencies
- Remarks:
- Parental male were not treated and the exposure duration for parental female was not clear. Further more the study did not follow any specific guideline was not conducted to GLP standard. Postmortem or histopathological examinations were not performed.
- Qualifier:
- no guideline followed
- Principles of method if other than guideline:
- To determine if sulfate in drinking water affects the reproductive performance of female mice during gestation and lactation over two parities. Sixty random-bred albino mice were assigned to six sulfate treatments. Sodium sulfate was added to deionized distilled water to give sulfate levels of: 0 (control), 0 (Na control); 625 ppm; 1250 ppm, 2500 ppm and 5,000. Treatments from 625 - 5000 ppm sulfate contained the same Na content (2,392 ppm) by varying the amount of Na bicarbonate content.
Parameters evaluated were water consumption, litter size, litter weaning weight, gestational and lactational weight gain of dams. - GLP compliance:
- no
- Limit test:
- no
- Justification for study design:
- - Premating exposure duration for parental (P0) animals: Treated water was available ad libitum beginning 1 week prior to breeding and was continued throughout the experiment.
- Basis for dose level selection: Selected dose levels of sulfate up to 5000 ppm.
- Inclusion/exclusion of extension of Cohorts. No, non-standard study (1989) which does not meet current guideline requirements.
- Termination time for F2: Not stated
- Route of administration: Ad libitum via drinking water
- Other considerations:
Low number of mice per group which does not reflect current guidance requirements.
Limited reproductive parameters recorded which does not reflect current guidance requirements.
The mouse is considered an applicable experimental model for the sow. - Specific details on test material used for the study:
- Sulfate in the form of reagent grade sodium sulfate. Sodium concentration was determined by varying the amount of sodium bicarbonate added.
- Species:
- mouse
- Strain:
- ICR
- Remarks:
- 60 random bred ICR virgin female mice were used.
- Details on species / strain selection:
- Outbred strain of albino mouse purchased from Harlan Sprague Dawley, Inc. Indianapolis, IN , USA.
- Sex:
- female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Harlan Sprague Dawley, Inc. Indianapolis (IN), USA.
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: Not stated
- Weight at study initiation: Not stated
- Fasting period before study: Not stated
- Housing: Housed individually in stainless steel cages with bedding
- Diet (ad libitum): yes, Commercially prepared breeders block (Wayne Mouse Breeder Blox, Wayne Pet Food Division, Continental Grain Co. Chicago ( IL), USA)
- Water (ad libitum): yes
- Acclimation period: 1 week
ENVIRONMENTAL CONDITIONS
- Temperature (°C): Environmentally controlled, ambient temperature 23.6°C
- Humidity (%): Environmentally controlled, ambient humidity 55%
- Air changes (per hr): Not stated
- Photoperiod (hrs dark / hrs light): 12-Hrs light/12-Hrs dark
IN-LIFE DATES: From: To: Not stated - Route of administration:
- oral: drinking water
- Vehicle:
- water
- Remarks:
- Deionized, distilled water
- Details on exposure:
- Exposure period: 1 week prior to breeding until study termination
- Details on mating procedure:
- After 1 weeks acclimation to the treatments, a male mouse that had received tap water was paired randomly with each female mouse. The females were checked every 24 h in the morning for the presence of a vaginal plug. If one was observed, the male mouse was removed and the female weighed.
- Analytical verification of doses or concentrations:
- no
- Duration of treatment / exposure:
- 1). Premating exposure of parental females to treated water was for 1 week prior to breeding.
2). There was no premating exposure period for males used for mating, on confirmation of mating the respective male removed from the cage.
3). All parental females dosed throughout until study termination
4): Each offspring was treated from day 21 post-partum until study termination - Frequency of treatment:
- Daily, drinking water containing sulfate was available ad libitum
- Details on study schedule:
- After 1 week of treatment acclimation to parental females a male mouse having received tap water was paired randomly with each female mouse. The females were checked every 24h (morning) for the presence of a vaginal plug. If observed, the male mouse was removed and the female weighed. Water consumption was measured daily, during the 2nd and 3rd week of gestation and the 1st and 2nd week of lactation.
Measurements of water consumption during week 1 of gestation were omitted due to excessive drinking bottle leakage. Water consumptions during week 3 of lactation were omitted because the pups could begin drinking water and interfere with accurate measurement of female
water consumption. At parturition, dams were weighed and litter size recorded. The litters then were standardized to eight pups per litter. At day 21 postpartum, the pups were weaned and the litters and dams were weighed individually. The dams then were rebred at first estrus immediately following weaning. This procedure was carried out over two parities. Only animals that whelped during each parity were used in the analysis. - Dose / conc.:
- 0 ppm (nominal)
- Remarks:
- Control (deionized, distilled water)
- Dose / conc.:
- 0 ppm (nominal)
- Remarks:
- Sodium control (in deionised, distilled water)
- Dose / conc.:
- 625 ppm (nominal)
- Remarks:
- Sulphate (Low) treatment group
- Dose / conc.:
- 1 250 ppm (nominal)
- Remarks:
- Sulfate treatment group
- Dose / conc.:
- 2 500 ppm (nominal)
- Remarks:
- Sulfate treatment group
- Dose / conc.:
- 5 000 ppm (nominal)
- Remarks:
- Sulfate (High) treatment group
- No. of animals per sex per dose:
- First parity:
Control : 6 dams
Sodium control: 6 dams
625 ppm: 4 dams (Low dose)
1250 ppm: 5 dams
2500 ppm: 9 dams
5000 ppm: 7 dams (High dose)
Second parity:
Control : 4 dams
Sodium control: 4 dams
625 ppm: 4 dams (Low dose)
1250 ppm: 4 dams
2500 ppm: 4 dams
5000 ppm: 4 dams (High dose) - Control animals:
- yes, concurrent no treatment
- yes, concurrent vehicle
- Details on study design:
- - Dose selection rationale: Six sulphate groups (treatment groups 2-6 contained the same Na content (i.e. 2392 ppm, by varying the amount of added sodium bicarbonate)
1: Control: Deionised water - 0 ppm
2: Sodium control: Deionised water - 0 ppm
3: Sulfate - 625 ppm
4: Sulfate - 1250 ppm
5: Sulfate - 2500 ppm
6: Sulfate - 5000 ppm
- Rationale for animal assignment: Random
- Fasting period before blood sampling for clinical biochemistry: Not stated - Parental animals: Observations and examinations:
- CAGE SIDE OBSERVATIONS: No
DETAILED CLINICAL OBSERVATIONS: No
BODY WEIGHT: Yes
At parturition, dams were weighed and litter size recorded.
WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
Water consumption was measured daily (graduated cylinder), during the 2nd and 3rd week of gestation and the 1st and 2nd week of lactation. Measurements of water consumption during week 1 of gestation were omitted due to excessive water leakage. Water consumptions during week 3 of lactation were omitted because the pups began drinking water and thus interfere with accurate measurement of female water consumption. - Oestrous cyclicity (parental animals):
- Dams were rebred at first estrus immediately following weaning.
- Litter observations:
- At parturition, dams were weighed and litter size was recorded. The litters then were standardized to eight pups/litter. Dams that whelped fewer than eight pups received pups from dams with greater than eight pups within the same treatment. When this was not possible, pups were cross-fostered pups from dams that were on a lower sulfate treatment. At 21 d post-partum, the pups
were weaned and the litters and dams were weighed individually. The dams then were rebred at first estrus immediately following weaning. This procedure was carried out over two parities. Only animals that whelped during each parity were used in the analysis.
During both parities, the litter size of live pups equalled the total number born as no mortalities at birth were detected.
During both parities, there were no differences (P > .10) in the litter size among treatments. The 21-d litter weaning weight for both parities were not affected (P > .10) by the sulfate treatment. A
quadratic response (P < .005) in weaning weight was observed within sulfate treatments during the second parity. - Postmortem examinations (parental animals):
- SACRIFICED: Not stated
GROSS NECROPSY: Not performed
HISTOPATHOLOGY / ORGAN WEIGHTS: Not performed - Postmortem examinations (offspring):
- SACRIFICED: Not stated
GROSS NECROPSY: Not performed
HISTOPATHOLOGY / ORGAN WEIGHTS: Not performed - Statistics:
- The least squares mean analysis of variance technique (SAS, 1982) was used to analyze the
data. One contrast was used to compare the deionized, distilled water control treatment to
all sulfate treatments. Sulfate treatment (2 to 6) effects were partitioned into linear, quadratic
and cubic components using orthogonal contrasts. Student's t-test (Steel and Torrie, 1980)
were used to determine the difference in water consumption between weeks in both gestation
and lactation.
Cited statistical references:
1: SAS (1982). SAS User's Guide: Statistics. SAS Inst., Inc., Cary, NC.
2: Steel, R.G.D. and J. H. Torrie. (1980). Principles and Procedures of Statistics. McGraw Hill Book Co., St. Louis, MO. - Reproductive indices:
- No effect on reproductive performance when litter size and weaning weight are used as indicators of maximum reproductive performance. Sulfates in the water up to 5000 ppm (ad libitum) do not affect reproductive performance in the gestating mouse.
- Offspring viability indices:
- During both parities, the litter size of live pups equalled the total number born as no mortalities at birth were detected.
- Clinical signs:
- no effects observed
- Mortality:
- no mortality observed
- Body weight and weight changes:
- effects observed, non-treatment-related
- Reproductive function: oestrous cycle:
- no effects observed
- Reproductive function: sperm measures:
- not examined
- Reproductive performance:
- effects observed, non-treatment-related
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- 5 000 ppm
- Based on:
- test mat.
- Sex:
- female
- Remarks on result:
- not determinable because of methodological limitations
- Key result
- Critical effects observed:
- not specified
- Clinical signs:
- no effects observed
- Mortality / viability:
- no mortality observed
- Body weight and weight changes:
- effects observed, non-treatment-related
- Food consumption and compound intake (if feeding study):
- not examined
- Food efficiency:
- not examined
- Water consumption and compound intake (if drinking water study):
- effects observed, non-treatment-related
- Ophthalmological findings:
- not examined
- Haematological findings:
- not examined
- Clinical biochemistry findings:
- no effects observed
- Urinalysis findings:
- not examined
- Sexual maturation:
- not examined
- Anogenital distance (AGD):
- not examined
- Nipple retention in male pups:
- not examined
- Organ weight findings including organ / body weight ratios:
- not examined
- Gross pathological findings:
- not examined
- Histopathological findings:
- not examined
- Other effects:
- not specified
- Behaviour (functional findings):
- not examined
- Developmental immunotoxicity:
- not examined
- Key result
- Dose descriptor:
- NOAEL
- Generation:
- F1
- Effect level:
- 5 000 ppm
- Based on:
- test mat.
- Sex:
- male/female
- Remarks on result:
- not determinable because of methodological limitations
- Key result
- Critical effects observed:
- not specified
- Key result
- Reproductive effects observed:
- not specified
- Conclusions:
- No effect on reproductive performance was observed in female mice following the ad libitum ingestion of sulfate (from sodium sulfate) in drinking water at up to 5000 ppm during gestation or lactation over two parities.
- Executive summary:
In this study, six treatment groups of 10 female mice were administered water (ad libitum) as either: (0)Control (deionized water), (0)Sodium control (in deionised water) or 625 ppm, 1250 ppm, 2500 ppm or 5000 ppm sulfate. Concentrations were adapted by the addition of sodium bicarbonate content. Premating exposure of parental females to treated water was for 1 week prior to the addition of males used for mating, on confirmation of mating males were removed. All parental females were dosed throughout until study termination. Each offspring was treated from day 21 post-partum until study termination. No effect on reproductive performance were observed in female mice during gestation or lactation over two parities. Sulfate (from sodium sulfate) administered at concentrations up to 5000 ppm (ad libitum in drinking water) caused no effect on litter size, litter weaning weight, or gestational or lactational weight gain of dams over two parities. Gestational and lactational gain of dams
was not affected by level of sulfate consumed. Minor decreases in water consumption with increasing treatment (compared to the sodium control group) were noted during both parities. Also, weight gain in dams was greater during the first parity and a quadratic response to sulfates in litter weaning weights was observed in the second parity. There were no adverse effects on reproductive performance.
Referenceopen allclose all
entire duration of the study.
Mean food consumption was not affected by the treatment with the test item at any dose level.
In order of ascending dose levels, the overall differences in food consumption were: ±0.0%,
+2.4% and -0.8% during pre-pairing period and -1.7%, -7.4% and -2.9% during the after pairing
period (percentages refer to the respective values of the control group).
No statistically significant alterations of mean food consumption were observed at any dose level
when compared to the respective values in the control group.
In order of ascending dose levels, the overall differences in food consumption were: +6.0%,
±0.0% and -3.3% during the pre-pairing period, +1.6%, +2.0% and -3.6% during the gestation
period and -3.7%, +4.0% and -5.6% during the lactation period (percentages refer to the
respective values of the control group).
No significant changes were observed in mean body weight and mean body weight gain.
In the order of ascending dose levels, the overall mean body weight gains were: +11%, +9%,
+13% and +11% during the pre-pairing period, +6%, +5%, +5% and +4% during the pairing
period and +4%, +4%, +4% and +3% during the after pairing period (percentages refer to the
respective time intervals).
No significant alterations of mean body weights and mean body weight gain were observed at
any dose level when compared to the respective values in the control group.
In the order of ascending dose levels, the overall mean body weight gain was +10%, +11%,
+12% and +9% during the pre-pairing period and +57%, +59%, +57% and +57% during the
gestation period and +5%, +4%, +6% and +6% during the lactation (percentages refer to the
respective time intervals).
The median and mean precoital times were unaffected by treatment with the test item. Mean
precoital times were 3.3, 4.3, 3.3 and 3.1 days in order of ascending dose level. The median
precoital time was 3, 4, 3 and 3 days in order of ascending dose level.
Two females each in groups 1 and 2 were not pregnant. Thus the fertility indices were 80.0%,
80.0%, 100.0% and 100.0% in groups 1, 2, 3 and 4.
The mean duration of gestation was unaffected by treatment with the test item. Mean duration of
gestation was 21.4, 21.5, 21.7 and 21.5 days, in order of ascending dose level.
The mean number of corpora lutea per dam (determined at necropsy) was similar in all groups
(14.6, 14.1, 14.2 and 14.1 in order of ascending dose level) and gave no indication of a test item related effect.
The mean number of implantations per dam and the post-implantation losses were unaffected by
treatment with the test item.
The mean numbers of implantations per litter were 13.3, 13.5, 14.0 and 13.3 in order of
ascending dose level. The mean incidence of post-implantation loss as a percentage of total
implantations was 10.4, 5.6, 10.0 and 3.8% in order of ascending dose level.
The mean number of implantations per dam and the post-implantation losses were unaffected by
treatment with the test item.
The mean numbers of implantations per litter were 13.3, 13.5, 14.0 and 13.3 in order of
ascending dose level. The mean incidence of post-implantation loss as a percentage of total
implantations was 10.4, 5.6, 10.0 and 3.8% in order of ascending dose level.
The total number of pup loss during the first four days was only one pup in the control group on
day 3. Therefore the viability indices were 98.9% in the control group and 100.0%, in groups 2,
3 and 4.
In males, weights (absolute and relative to body weight) of testes and epididymides were not
affected by the treatment with the test item in any groups.
found bilaterally and this finding was considered to be associated with their infertility. In the
above 2 animals, aspermia and cellular debris in epididymides which were considered to be
associated with the tubular atrophy of testes were also observed. No microscopic findings were
found in other reproductive organs of these animals, reproductive organs of other infertile males
(nos. 8 and 16) and ovaries of females (nos. 48, 49, 56 and 59) that did not give birth.
Treatment with the test item did not reveal effects on the completeness of stages or cell
populations. There was no indication for maturation arrest or any other degenerative type.
on mouth in one pup in group 1, in four pups in group 3 and one pup in group 4 and hind foot
injured and toes missing in one pup in group 2. None of these findings gave an indication of a
test item-related effect.
Sex ratios at first litter check and on day 4 post partum were unaffected by exposure to the test
item. The proportion of males on day 4 post partum was 48%, 49%, 50% and 45% in order of
ascending dose level.
Mean pup weights were unaffected by treatment with the test item. On day 1 post partum mean
pup weights were 6.1, 6.1, 6.4 and 6.2 g for combined data of male and female pups in order of
ascending dose level. Mean pup weight development during lactation was unaffected by treatment with the test item. Mean pup weights on day 4 post partum were 9.6, 9.4, 9.7 and 9.3 g for combined data of male and female pups in order of ascending dose level.
No abnormal findings were noted at macroscopic examination of the pups.
Table 1. Summary of food consumption in P0 male.
PRE-PAIRING PERIOD Days 1-8 MEAN ST.DEV. N Days 8-14 MEAN ST.DEV. N MEAN OF MEANS Over PRE-PAIRING PERIOD AFTER PAIRING PERIOD Days 1-6 MEAN ST.DEV. N |
Group 1 mg/kg 24.8 1.0 10 25.7 1.6 10 25.3 24.2 10 |
Group 2 mg/kg 1.7 10 1.6 10 25.3 2.2 10 |
Group 3 mg/kg 1.5 10 2.0 10 25.9 2.8 10 |
Group 4 mg/kg 2.1 10 1.7 10 25.1 1.4 10 |
Table 2. Summary of food consuption P0 female
PRE-PAIRING PERIOD Days 1-8 MEAN ST.DEV. N Days 8-14 MEAN ST.DEV. N MEAN OF MEANS Over PRE-PAIRING PERIOD GESTATION PERIOD Days 0-7 MEAN ST.DEV. N Days 7-14 MEAN ST.DEV. N Days 14-21 MEAN ST.DEV. N |
Group 1 0 mg/kg 17.2 1.2 10 19.3 1.4 10 18.2 22.9 1.8 8 24.9 2.1 8 26.5 2.1 8 |
Group 2 100 mg/kg 18.2 1.4 10 20.3 1.2 10 19.3 22.6 4.0 8 25.6 1.4 8 27.6 1.5 8 25.2 31.1 2.7 8 |
Group 3 300 mg/kg 17.2 1.6 10 19.1 1.8 10 18.2 22.9 1.5 10 25.1 1.8 10 27.7 2.6 10 25.3 33.6 4.4 10 |
Group 4 1000 mg/kg 16.9 1.0 10 18.4 1.3 10 17.6 21.6 1.4 10 23.5 1.6 10 26.6 1.9 10 23.9 30.5 4.3 10 |
MEAN OF MEANS 24.8 Over GESTATION PERIOD (Days 0-21) LACTATION PERIOD Days 1-4 MEAN 32.3 ST.DEV. 5.5 N 8 |
*/**/- : DUNNETT-Test based on pooled variance sig. at 5% (*), 1% (**) or not sig. (-)
Table 3. summary of differences in mean food consumption of P0 males
PRE-PAIRING PERIOD Days 1-8 MEAN ST.DEV. N DIFF (%)* Days 8-14 MEAN ST.DEV. N DIFF (%)* MEAN OF MEANS Over PRE-PAIRING PERIOD DIFF (%)* Over PRE-PAIRING PERIOD AFTER PAIRING PERIOD Days 1-6 MEAN ST.DEV. N DIFF (%)* |
Group 1 mg/kg 24.8 1.0 --- 25.7 1.6 --- --- 24.2 1.1 10 --- |
Group 2 mg/kg 25.1 1.7 10 +1.2 25.4 1.6 10 -1.2 ±0.0 23.8 2.2 -1.7 |
Group 3 mg/kg 25.7 1.5 10 +3.6 26.1 2.0 10 +1.6 +2.4 22.4 2.8 -7.4 |
Group 4 mg/kg 24.8 2.1 10 ±0.0 25.4 1.7 10 -1.2 -0.8 23.5 1.4 -2.9 |
*: Percentage relative to the control group
Table 4. Summary of diffrenecs in mean food consumption in P0 Female
PRE-PAIRING PERIOD Days 1-8 MEAN ST.DEV. N DIFF (%)* Days 8-14 MEAN ST.DEV. N DIFF (%)* MEAN OF MEANS Over PRE-PAIRING PERIOD DIFF (%)* Over PRE-PAIRING PERIOD GESTATION PERIOD Days 0-7 MEAN ST.DEV. N DIFF (%)* Days 7-14 MEAN ST.DEV. N DIFF (%)* Days 14-21 MEAN ST.DEV. N DIFF (%)* |
Group 1 mg/kg 17.2 1.2 --- 19.3 1.4 --- --- 22.9 1.8 --- 24.9 2.1 --- 26.5 2.1 8 --- |
Group 2 mg/kg 18.2 1.4 10 +5.8 20.3 1.2 10 +5.2 +6.0 22.6 4.0 8 -1.3 25.6 1.4 8 +2.8 27.6 1.5 8 +4.2 25.2 |
Group 3 mg/kg 17.2 1.6 10 ±0.0 19.1 1.8 10 -1.0 ±0.0 22.9 1.5 10 ±0.0 25.1 1.8 10 +0.8 27.7 2.6 10 +4.5 25.3 |
Group 4 mg/kg 16.9 1.0 10 -1.7 18.4 1.3 10 -4.7 -3.3 21.6 1.4 10 -5.7 23.5 1.6 10 -5.6 26.6 1.9 10 +0.4 23.9 |
MEAN OF MEANS 24.8 |
*: Percentage relative to the control group
Table 5. Summary of P0 male body weight gain.
|
Pre-pairing period |
Group 1 (0 mg/kg bw/day) |
Group 2 (100 mg/kg bw/day) |
Group 3 (300 mg/kg bw/day) |
Group 4 ( 1000 mg/kg bw/day) |
|||||
|
Day 1 MEAN ST.DEV. N Day 2 MEAN ST.DEV. N Day 3 MEAN ST.DEV. N Day 4 MEAN ST.DEV. N Day 5 MEAN ST.DEV. N Day 6 MEAN ST.DEV. N Day 7 MEAN ST.DEV. N Day 8 MEAN ST.DEV. N Day 9 MEAN ST.DEV. N Day 10 MEAN ST.DEV. N Day 11 MEAN ST.DEV. N |
308 9 10 314 9 10 317 9 10 321 10 10 318 10 10 326 11 10 322 10 10 326 10 10 330 10 10 335 11 10 339 10 10 |
307 - 6 10 313 - 6 10 315 - 7 10 319 - 8 10 317 - 7 10 324 - 8 10 320 - 8 10 323 - 9 10 327 - 9 10 329 - 10 10 332 - 10 10 |
310 - 8 10 318 - 9 10 319 - 9 10 325 - 9 10 324 - 9 10 331 - 10 10 329 - 11 10 334 - 11 10 336 - 14 10 340 - 13 10 344 - 12 10 |
307 - 9 10 317 - 10 10 317 - 9 10 320 - 11 10 319 - 9 10 326 - 10 10 323 - 10 10 328 - 11 10 332 - 12 10 335 - 12 10 338 - 12 10 |
|||||
|
PRE-PAIRING PERIOD Day 12 MEAN ST.DEV. N Day 13 MEAN ST.DEV. N Day 14 MEAN ST.DEV. N PAIRING PERIOD Day 1 MEAN ST.DEV. N Day 2 MEAN ST.DEV. N Day 3 MEAN ST.DEV. N Day 4 MEAN ST.DEV. N Day 5 MEAN ST.DEV. N Day 6 MEAN ST.DEV. N Day 7 MEAN ST.DEV. N Day 8 MEAN ST.DEV. N |
338 11 10 339 12 10 344 11 10 337 14 10 339 12 10 342 12 10 344 11 10 347 12 10 351 11 10 347 12 10 350 12 10 |
332 - 12 10 332 - 12 10 335 - 12 10 331 - 14 10 332 - 15 10 335 - 15 10 336 - 17 10 339 - 17 10 342 - 17 10 338 - 18 10 343 - 17 10 |
345 - 13 10 346 - 14 10 350 - 14 10 345 - 15 10 348 - 14 10 350 - 17 10 352 - 15 10 355 - 15 10 357 - 15 10 356 - 16 10 359 - 17 10 |
338 - 13 10 339 - 13 10 342 - 13 10 338 - 14 10 340 - 14 10 341 - 12 10 343 - 13 10 347 - 12 10 347 - 12 10 344 - 13 10 347 - 12 10 |
|||||
Day 9 MEAN ST.DEV. N Day 10 MEAN ST.DEV. N AFTER PAIRING PERIOD Day 1 MEAN ST.DEV. N Day 2 MEAN ST.DEV. N Day 3 MEAN ST.DEV. N Day 4 MEAN ST.DEV. N Day 5 MEAN ST.DEV. N Day 6 MEAN ST.DEV. N |
354 12 10 356 12 10 356 14 10 361 13 10 362 13 10 360 14 10 362 14 10 371 14 10 |
345 - 17 10 348 - 17 10 349 - 19 10 353 - 19 10 355 - 19 10 351 - 19 10 356 - 19 10 362 - 20 10 |
361 - 18 10 364 - 17 10 366 - 19 10 370 - 19 10 371 - 19 10 370 - 19 10 373 - 19 10 380 - 19 10 |
349 - 12 10 352 - 12 10 351 - 12 10 355 - 13 10 356 - 13 10 355 - 13 10 358 - 13 10 363 - 14 10 |
|
Table 6. Summary of Female body weight gain
Periods |
Group 1 (0 mg/kg bw/day) |
Group 2 (100 mg/kg bw/day) |
Group 3 (300 mg/kg bw/day) |
Group 4 (1000 mg/kg bw/day) |
||||||||
|
Pre-pairing period
Day 1 MEAN ST.DEV. N Day 2 MEAN ST.DEV. N Day 3 MEAN ST.DEV. N Day 4 MEAN ST.DEV. N Day 5 MEAN ST.DEV. N Day 6 MEAN ST.DEV. N Day 7 MEAN ST.DEV. N Day 8 MEAN ST.DEV. N Day 9 MEAN ST.DEV. N Day 10 MEAN ST.DEV. N Day 11 MEAN ST.DEV. N |
201 8 10 205 7 10 203 7 10 207 8 10 207 9 10 209 8 10 207 8 10 212 10 10 214 10 10 214 8 10 215 8 10 |
198 - 11 10 202 - 9 10 204 - 12 10 207 - 15 10 206 - 12 10 208 - 10 10 208 - 13 10 211 - 13 10 213 - 13 10 214 - 11 10 219 - 13 10 |
198 - 12 10 203 - 15 10 204 - 13 10 208 - 14 10 205 - 13 10 208 - 15 10 209 - 13 10 213 - 12 10 214 - 13 10 214 - 16 10 221 - 14 10 |
202 - 9 10 206 - 9 10 208 - 9 10 211 - 9 10 208 - 9 10 212 - 10 10 211 - 9 10 214 - 11 10 215 - 12 10 215 - 11 10 221 - 11 10 |
|||||||
|
Day 12 MEAN ST.DEV. N Day 13 MEAN ST.DEV. N Day 14 MEAN ST.DEV. N PAIRING PERIOD Day 1 MEAN ST.DEV. N Day 2 MEAN ST.DEV. N Day 3 MEAN ST.DEV. N Day 4 MEAN ST.DEV. N Day 5 MEAN ST.DEV. N Day 6 MEAN ST.DEV. N Day 7 MEAN ST.DEV. N Day 8 MEAN ST.DEV. N |
219 9 10 221 10 10 221 9 10 227 3 6 228 6 6 236 4 3 238 8 2 249 0 1 251 0 1 253 0 1 258 0 1 |
218 - 15 10 220 - 13 10 219 - 11 10 226 - 13 10 232 - 11 7 235 - 14 6 235 - 1 2 240 - 1 2 239 - 6 2 243 0 1 245 0 1 |
221 - 14 10 220 - 13 10 221 - 16 10 228 - 13 9 231 - 13 8 230 - 16 3 246 - 0 1 249 - 0 1 250 - 0 1 --- --- 0 --- --- 0 |
221 - 12 10 219 - 12 10 220 - 12 10 228 - 11 10 226 - 12 9 226 - 18 3 --- --- 0 --- --- 0 --- --- 0 --- --- 0 --- --- 0 |
|
||||||
Pairing period
Day 9 MEAN ST.DEV. N Day 10 MEAN ST.DEV. N GESTATION PERIOD Day 0 MEAN ST.DEV. N Day 1 MEAN ST.DEV. N Day 2 MEAN ST.DEV. N Day 3 MEAN ST.DEV. N Day 4 MEAN ST.DEV. N Day 5 MEAN ST.DEV. N Day 6 MEAN ST.DEV. N Day 7 MEAN ST.DEV. N Day 8 MEAN ST.DEV. N |
263 0 1 265 0 1 227 11 8 231 12 8 235 11 8 239 11 8 244 11 8 247 11 8 249 14 8 253 13 8 258 14 8 |
252 0 1 251 0 1 228 - 16 8 234 - 17 8 240 - 17 8 242 - 16 8 243 - 17 8 249 - 17 8 254 - 17 8 257 - 15 8 263 - 16 8 |
--- --- 0 --- --- 0 228 - 14 10 234 - 13 10 240 - 14 10 244 - 15 10 248 - 14 10 252 - 13 10 256 - 13 10 258 - 13 10 264 - 13 10 |
--- --- 0 --- --- 0 227 - 11 10 232 - 13 10 237 - 13 10 239 - 12 10 241 - 11 10 245 - 11 10 249 - 12 10 253 - 12 10 256 - 12 10 |
|
|||||||
|
Day 20 MEAN ST.DEV. N Day 21 MEAN ST.DEV. N LACTATION PERIOD Day 1 MEAN ST.DEV. N Day 2 MEAN ST.DEV. N Day 3 MEAN ST.DEV. N Day 4 MEAN ST.DEV. N |
352 19 8 362 23 8 273 12 8 275 13 8 280 14 8 286 13 8 |
359 - 22 8 373 - 26 8 278 - 18 8 279 - 18 8 281 - 15 8 290 - 19 8 |
357 - 30 10 369 - 31 10 278 - 15 10 288 - 19 10 285 - 14 10 295 - 18 10 |
349 - 16 10 364 - 18 10 267 - 11 10 272 - 13 10 277 - 14 10 284 - 11 10 |
|
*/**/- : DUNNETT-Test based on pooled variance sig. at 5% (*), 1% (**) or not sig. (-)
Table 7. Mating performance
Days of the pairing period. |
Group 1 mg/kg
|
Group 2 mg/kg
|
Group 3 mg/kg
|
Group 4 mg/kg
|
1 2 3 4 5 7 11 |
4 -
3 1 1 - 1 |
- 3
1 4 - 1 1 |
1 1 5 2 - 1 - |
- 1 7 2 - - - - |
Median precoital time |
3 |
4 |
3 |
3 |
Mean precoital time |
3.3 |
4.3 |
3.3 |
3.1 |
N |
10 |
10 |
10 |
10 |
Table 8. Fertility parameters
Days of Parameters |
Group 1 mg/kg
|
Group 2 mg/kg
|
Group 3 mg/kg
|
Group 4 mg/kg
|
Mating (%) |
100 |
100 |
100 |
100 |
Fertility index (%) |
80 |
80 |
100 |
100 |
Conception index (%) |
80 |
80 |
100 |
1000 |
Gestation index – Breeding (%) |
100 |
100 |
100 |
100 |
Percentage mating = (Females mated / Females paired) * 100
Fertility index = (Females achieving a pregnancy / Females paired) * 100
Conception rate = (Females achieving a pregnancy / Females mated) * 100
Gestation index = (Number of females with living pups / Number of
females pregnant) * 100
Table 9. Summary of mean breading data per group.
Parameters |
Group 1 mg/kg
|
Group 2 mg/kg
|
Group 3 mg/kg
|
Group 4 mg/kg
|
Total Litters Mean duration of gestation |
8
21.4 |
8
21.5 |
10
21.7 |
10
21.5 |
Mean implantation |
13.3 |
13.5 |
14.0 |
13.3 |
Post implantation loss |
1.4 |
0.8 |
1.4 |
0.5 |
Dead pups at first litter check |
0.3 |
0.3 |
0.0 |
0.0 |
Live pups at first check |
11.9 |
12.8 |
12.6 |
12.8 |
Postnatal loss days 0- 4 |
0.1 |
0.0 |
0.0 |
0.0 |
Live pups day 4 |
11.8 |
12.8 |
12.6 |
12.8 |
Birth index (#) |
89.6 |
94.4 |
90.0 |
96.2# |
Viability index (#) |
98.9 |
100 |
100 |
100 |
# / ## : Fisher's Exact Test significant at 5% (#) or 1% (##) level
Birth index = (number of pups born alive / number of implantations) * 100
Viability index = (number of alive pups on day 4 p.p. / number of pups born alive) * 100
Table 9. summary of organ/bosy weight ratio and percentage mean values.
Organs |
Group 1 (0 mg/kg bw/day
|
Group 2 (100 mg/kg bw/day
|
Group 3 (300 mg/kg b/day
|
Group 4 (1000 mg/kg bw/day)
|
Body weight
% |
366.7
14.4 |
357.3
20.6 |
373.8
18.6 |
360.8
13.5 |
Testis (L)
% |
1.78
0.49 |
1.72
0.48 |
2.00
0.54 |
1.92
0.53 |
Testis (R)
% |
1.75
0.48 |
1.67
0.47 |
1.92
0.52 |
1.92
0.53 |
Epididi (R)
% |
0.681
0.186 |
0.642
0.180 |
0.730
0.195
|
0.714
0.198 |
Epidid (L)
% |
0.675
0.185 |
0.644
0.180 |
0.743
0.199 |
0.738
0.205 |
or weight of pigs at birth or at weaning were observed. Fecal consistency was normal in all treatments. Water consumption did not differ during gestation but
increased during lactation as salt level increased. Results suggest that sulfates up to 3320 ppm in water have no significant effect on reproduction in the gilt or sow.
or feed to gain ratio were noted. Water consumption was greater among pigs that received water with added sulfates. No differences were observed in pigs that received water containing sodium sulfate or equal parts of sulfate from sodium and magnesium sulfate.
Effect Of Sulfate Content Of Water On Reproductive Performance
Parameter | 0(Control) | 3000 ppm | 3000 ppm |
No. of pigs a | 16 | 18 | 17 |
Avg initial wt. (kg) | 7.5 | 8.0 | 7.7 |
Avg final wt. (kg) | 13.4 | 15.0 | 13.8 |
Avg daily gain (kg) | 0.21 | 0.25 | 0.22 |
Feed to gain ratio | 2.25 | 2.05 | 2.18 |
Avg daily water consumption (l) | 1.25 b | 1.89 c | 1.63 d |
Avg fecal condition e | 1.7 f | 3.3 g | 3.6 g |
a: Three replications of six pigs per treatment. Three pigs died, data not included.
b, c, d: Means on the same line with different superscripts are significantly different (P<.05).
e: Based on a score of 1 to 5, with 1 being firm.
f, g: Means on same line with different superscripts are significantly different (P<.01).
or weight of pigs at birth or at weaning were observed. Fecal consistency was normal in all treatments. Water consumption did not differ during gestation but
increased during lactation as salt level increased. Results suggest that sulfates up to 3320 ppm in water have no significant effect on reproduction in the gilt or sow.
or feed to gain ratio were noted. Water consumption was greater among pigs that received water with added sulfates. No differences were observed in pigs that received water containing sodium sulfate or equal parts of sulfate from sodium and magnesium sulfate.
Effect Of Sulfate Content Of Water On Reproductive Performance
Parameter | 0(Control) | 3000 ppm | 3000 ppm |
No. of pigs a | 16 | 18 | 17 |
Avg initial wt. (kg) | 7.5 | 8.0 | 7.7 |
Avg final wt. (kg) | 13.4 | 15.0 | 13.8 |
Avg daily gain (kg) | 0.21 | 0.25 | 0.22 |
Feed to gain ratio | 2.25 | 2.05 | 2.18 |
Avg daily water consumption (l) | 1.25 b | 1.89 c | 1.63 d |
Avg fecal condition e | 1.7 f | 3.3 g | 3.6 g |
a: Three replications of six pigs per treatment. Three pigs died, data not included.
b, c, d: Means on the same line with different superscripts are significantly different (P<.05).
e: Based on a score of 1 to 5, with 1 being firm.
f, g: Means on same line with different superscripts are significantly different (P<.01).
There was no differences in litter size between treatments and the 21-d litter weaning weight were not affected.
Water consumption during both weeks of gestation and both weeks of lactation were observed with no effect on reproductive performance of female mice.
There was no differences in litter size between treatments and the 21-d litter weaning weight were not affected. A quadratic response in weaning weight was however observed within treatment groups during the second parity.
Water consumption during both weeks of gestation and both weeks of lactation were observed with no effect on reproductive performance of female mice.
Effect Of Sulfate Treatment On Litter Sizea At Weaning And Litter Weaning Weight (Both Parities)
Sulfate(ppm) | First parityb | Second parityb | ||
Pups/ | Weaning wt.(g)c | Pups/ | Weaning wt.(g)c | |
0 (Control) Deionized, distilled water | 8.0 | 123.3 | 11.3 | 129.7 |
Na control ("0" sulfate) | 8.3 | 128.1 | 13.0 | 130.6 |
625 | 8.8 | 114.3 | 13.3 | 126.9 |
1250 | 7.8 | 130.0 | 13.5 | 141.2 |
2500 | 8.0 | 132.7 | 13.0 | 144.2 |
5000 | 8.5 | 125.9 | 15.5 | 130.4 |
SE | 0.6 | 6.2 | 1.3 | 3.8d |
a: Total number born and number born alive were identical because no mortality was detected at parturition.
b: Sulfate treatment means within columns do not differ from the O sulfate, Na control treatment (P > .10).
c: Weaning weights of the standardized (eight pups) litters.
d: Quadratic response within sulfate treatment (P <0.005).
There was no differences in litter size between treatments and the 21-d litter weaning weight were not affected.
Water consumption during both weeks of gestation and both weeks of lactation were observed with no effect on reproductive performance of female mice.
There was no differences in litter size between treatments and the 21-d litter weaning weight were not affected. A quadratic response in weaning weight was however observed within treatment groups during the second parity.
Water consumption during both weeks of gestation and both weeks of lactation were observed with no effect on reproductive performance of female mice.
Effect Of Sulfate Treatment On Litter Sizea At Weaning And Litter Weaning Weight (Both Parities)
Sulfate(ppm) | First parityb | Second parityb | ||
Pups/ | Weaning wt.(g)c | Pups/ | Weaning wt.(g)c | |
0 (Control) Deionized, distilled water | 8.0 | 123.3 | 11.3 | 129.7 |
Na control ("0" sulfate) | 8.3 | 128.1 | 13.0 | 130.6 |
625 | 8.8 | 114.3 | 13.3 | 126.9 |
1250 | 7.8 | 130.0 | 13.5 | 141.2 |
2500 | 8.0 | 132.7 | 13.0 | 144.2 |
5000 | 8.5 | 125.9 | 15.5 | 130.4 |
SE | 0.6 | 6.2 | 1.3 | 3.8d |
a: Total number born and number born alive were identical because no mortality was detected at parturition.
b: Sulfate treatment means within columns do not differ from the O sulfate, Na control treatment (P > .10).
c: Weaning weights of the standardized (eight pups) litters.
d: Quadratic response within sulfate treatment (P <0.005).
Effect on fertility: via oral route
- Endpoint conclusion:
- no adverse effect observed
- Dose descriptor:
- NOAEL
- 1 000 mg/kg bw/day
- Species:
- rat
Effect on fertility: via inhalation route
- Endpoint conclusion:
- no study available
Effect on fertility: via dermal route
- Endpoint conclusion:
- no study available
Additional information
No relevant studies investigating the effects of sulphuric acid exposure in animals are available. The testing of sulphuric acid for effects on fertility in a conventional two-generation study in the rat is not proposed. The corrosive nature of the substance means that testing cannot be justified for reasons of animal welfare. The usual route of exposure in the two-generation reproductive toxicity study is not relevant to human exposure as effects will be dominated by local irritation of the gastrointestinal tract. Similarly testing of sulphuric acid using dermal exposure cannot be justified. In terms of worker exposure, the inhalation route is most relevant. However numerous inhalation toxicity studies performed in a number of different species and of various durations have not demonstrated any systemic effects of exposure at levels up to those causing mortality. The effects of inhalation exposure to sulphuric acid are limited to the respiratory tract (structure and function); effects on other organs and tissues have not been identified. Specifically, no gross effects on the reproductive organs were observed in two species (rat and guinea pig) following chronic exposure. Following a comprehensive literature review, the OECD SIDS for sulphuric acid (2001) concluded that sulphuric acid ‘...is not expected to be absorbed or distributed throughout the body. Therefore it is not likely that it will reach male and female reproductive organs following exposures by any route’. The ATSDR (1998) comes to a similar conclusion, stating that ‘Because sulfuric acid is a direct-acting toxicant and not likely to reach reproductive organs or the foetus, reproductive and developmental effects are unlikely to occur in humans exposed to sulfuric acid’. As previously discussed, sulphuric acid will immediately dissociate on contact with the body to form sulphate and hydrogen (hydronium) ions. The hydrogen ion is likely to be entirely responsible for the site of contact effects of sulphuric acid. Any hydrogen ion absorbed into the systemic circulation will be subject to the normal homeostatic mechanisms governing physiological pH, and will be excreted in the urine when in excess. Sulphate ions are also a normal product of metabolism and are present in significant amounts in the body. Studies have demonstrated the absorption of sulphate following inhalation exposure to sulphuric acid, however levels of sulphate are under homeostatic control and any ‘excess’ sulphate will be excreted in the urine. It is calculated that exposure to sulphuric acid at the TWA will result in negligible additional systemic sulphate exposure, which will not have any effect on fertility. It is therefore considered that the testing of sulphuric acid for effects on fertility cannot be justified, both on scientific grounds (due to the lack of systemic exposure) and for reasons of animal welfare (due to the corrosive nature of the substance.
A waiver is proposed for this endpoint based on information provided by Cecatelli (2010) “Sodium Sulphate: Reproduction/Developmental Toxicity Screening Test in the Han Wistar Rat” a reproduction/developmental toxicity screening test, Andres, C. J. and Cline, T. R. (1989) “Influence of sulfate in drinking water on mouse reproduction during two parities” evaluating toxicity to reproduction, and a study by Murray, F.J. et al (1979) “Embryotoxicity of inhaled sulfuric acid aerosol in mice and rabbits” evaluating developmental toxicity.
The study R. Ceccatelli 2010, is a valid investigation of the toxicological effects resulting from repeated oral-gavage administration of the test item Sodium Sulphate to rats. Sodium Sulphate was administered in highly purified water as vehicle at dosages of 100, 300, and 1000 mg/kg body weight/day, and controls received the vehicle only. Sodium Sulphate was administered to male rats for at least 28 days and to female rats for 14 days prior to pairing, through the pairing and gestation periods until the F1 generation reached day 4 post partum. In absence of any effect the general NOEL (No Observed Effect Level) was established at 1000 mg/kg/day. As no effects were observed, the NOEL (No Observed Effect Level) for reproduction/ developmental toxicity was considered to be 1000 mg/kg/day.
In the study by Andres, C. J. and Cline, T. R. (1989), six treatment groups of 10 female mice were administered water (ad libitum) as either: (0)Control (deionized water), (0)Sodium control (in deionized water) or 625 ppm, 1250 ppm, 2500 ppm or 5000 ppm sulfate in drinking water. Concentrations were adapted by the addition of sodium bicarbonate content. Premating exposure of parental females to treated water was for 1 week prior to the addition of males used for mating, on confirmation of mating males were removed. All parental females were dosed throughout until study termination. Each offspring was treated from day 21 post-partum until study termination. No effect on reproductive performance were observed in female mice during gestation or lactation over two parities. Sulfate (from sodium sulfate) administered at concentrations up to 5000 ppm (ad libitum in drinking water) caused no effect on litter size, litter weaning weight, or gestational or lactational weight gain of dams over two parities. Gestational and lactational gain of dams was not affected by level of sulfate consumed. Minor decreases in water consumption with increasing treatment (compared to the sodium control group) were noted during both parities. Also, weight gain in dams was greater during the first parity and a quadratic response to sulfates in litter weaning weights was observed in the second parity. There were no adverse effects of reproductive performance.
In the study by Murray, F..J. et al (1979), mated female mice were exposed (whole body) to atmospheres containing aerosols of sulphuric acid at 0, 5.7 mg/m3 (MMAD 1.6 um) or 19.3 mg/m3 (MMAD 2.4 um) on Days 6 -15 of gestation (mice). Animals were sacrificed at Days 18 (mice). Evidence of mild toxicity was seen in maternal mice (reduced food consumption) at 20 mg/m3. No evidence of teratogenicity or developmental toxicity was seen. Inseminated female rabbits were exposed (whole body) to atmospheres containing aerosols of sulphuric acid at 0, 5.7 mg/m3 (MMAD 1.6 um) or 19.3 mg/m3 (MMAD 2.4 um) on Days 6 -18 of gestation (rabbits). Animals were sacrificed at Day 29 of gestation (rabbits). Evidence of mild toxicity was seen in maternal rabbits (reduced weight gain, respiratory tract effects) at 20 mg/m3.No evidence of teratogenicity or developmental toxicity was seen.
Effects on developmental toxicity
Description of key information
A study of developmental toxicity in the rabbit and mouse (using inhalation exposure) has been performed with sulphuric acid, to a protocol broadly comparable to OECD 414: NOAEC of 19.3 mg/m3 for developmental toxicity in both species and 5.7 mg/m3 for maternal toxicity in both species (Murray et al, 1979).
Link to relevant study records
- Endpoint:
- developmental toxicity
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Literature study, comparable to OECD 414
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 414 (Prenatal Developmental Toxicity Study)
- Deviations:
- yes
- Remarks:
- Number of exposure levels
- Principles of method if other than guideline:
- Sulphuric acid was tested by inhalation for embryotoxicity in rabbits according to a protocol similar to OECD guideline 414
- GLP compliance:
- no
- Remarks:
- Older, published study
- Limit test:
- no
- Species:
- rabbit
- Strain:
- other: New Zealand White, CF-1
- Route of administration:
- inhalation: aerosol
- Type of inhalation exposure (if applicable):
- whole body
- Vehicle:
- water
- Analytical verification of doses or concentrations:
- yes
- Details on mating procedure:
- Nulliparous female rabbits were artificially inseminated. The day on which rabbits were artificially inseminated was considered day zero of gestation.
- Duration of treatment / exposure:
- 7 hours/day: Day 6-18 of gestation
- Frequency of treatment:
- Daily
- Duration of test:
- Day 29 of gestation
- No. of animals per sex per dose:
- 20 (with controls)
- Control animals:
- yes
- Dose descriptor:
- LOAEC
- Effect level:
- 19.3 mg/m³ air (analytical)
- Basis for effect level:
- other: Maternal toxicity
- Dose descriptor:
- NOAEC
- Effect level:
- 5.7 mg/m³ air (analytical)
- Basis for effect level:
- other: Maternal toxicity
- Dose descriptor:
- NOAEC
- Effect level:
- 19.3 mg/m³ air (analytical)
- Basis for effect level:
- other: Developmental toxicity
- Dose descriptor:
- NOAEC
- Effect level:
- 19.3 mg/m³ air (analytical)
- Basis for effect level:
- other: Teratogenicity
- Developmental effects observed:
- not specified
- Conclusions:
- No evidence of teratogenicity, embryotoxicity, foetotoxicity or developmental toxicity was seen under the conditions of this study, at exposure levels sufficient to cause mild maternal toxicity.
- Executive summary:
Inseminated female rabbits were exposed (whole body) to atmospheres containing aerosols of sulphuric acid at 0, 5.7 mg/m3 (MMAD 1.6 um) or 19.3 mg/m3 (MMAD 2.4 um) on Days 6 -18 of gestation (rabbits). Animals were sacrificed at Day 29 of gestation (rabbits). Evidence of mild toxicity was seen in maternal rabbits (reduced weight gain, respiratory tract effects) at 20 mg/m3. No evidence of teratogenicity or developmental toxicity was seen.
- Endpoint:
- developmental toxicity
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Literature study, comparable to OECD 414
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 414 (Prenatal Developmental Toxicity Study)
- Deviations:
- yes
- Remarks:
- Number of exposure levels
- Principles of method if other than guideline:
- Sulphuric acid was tested by inhalation for embryotoxicity in mice according to a protocol similar to OECD guideline 414
- GLP compliance:
- no
- Remarks:
- Older, published study
- Limit test:
- no
- Species:
- mouse
- Route of administration:
- inhalation: aerosol
- Type of inhalation exposure (if applicable):
- whole body
- Vehicle:
- water
- Analytical verification of doses or concentrations:
- yes
- Details on mating procedure:
- Nulliparous female mice were pen-bred with fertile males of the same strain. The day on which a vaginal plug was observed in mice was considered day zero of gestation.
- Duration of treatment / exposure:
- 7 hours/day: Day 6-15 of gestation
- Frequency of treatment:
- Daily
- Duration of test:
- Day 18 of gestation
- No. of animals per sex per dose:
- 35 (with controls)
- Control animals:
- yes
- Dose descriptor:
- LOAEC
- Effect level:
- 19.3 mg/m³ air (analytical)
- Basis for effect level:
- other: maternal toxicity
- Dose descriptor:
- NOAEC
- Effect level:
- 5.7 mg/m³ air (analytical)
- Basis for effect level:
- other: maternal toxicity
- Dose descriptor:
- NOAEC
- Effect level:
- 19.3 mg/m³ air (analytical)
- Basis for effect level:
- other: developmental toxicity
- Dose descriptor:
- NOAEC
- Effect level:
- 19.3 mg/m³ air (analytical)
- Basis for effect level:
- other: teratogenicity
- Abnormalities:
- not specified
- Developmental effects observed:
- not specified
- Conclusions:
- No evidence of teratogenicity, embryotoxicity, foetotoxicity or developmental toxicity was seen under the conditions of this study, at exposure levels sufficient to cause mild maternal toxicity.
- Executive summary:
Mated female mice were exposed (whole body) to atmospheres containing aerosols of sulphuric acid at 0, 5.7 mg/m3 (MMAD 1.6 um) or 19.3 mg/m3 (MMAD 2.4 um) on Days 6 -15 of gestation (mice). Animals were sacrificed at Days 18 (mice). Evidence of mild toxicity was seen in maternal mice (reduced food consumption) at 20 mg/m3. No evidence of teratogenicity or developmental toxicity was seen.
Referenceopen allclose all
Inhalation of 5 or 20 mg sulphuric acid/m3 did cause any signs of toxicity. The incidence of pregnancy was not significantly altered among rabbits. In rabbits, a statistically significant decrease in maternal bodyweight gain was noted during the first few days of exposure to 20 mg/m3. The liver weight of dams was unaltered by exposure to sulphuric acid in rabbits. Evidence of toxicity that could be attributed to sulphuric acid was observed in the respiratory tract in rabbits, as a trend toward a dose-related increase in the incidence of subacute rhinitis and tracheitis was noted. The mean numbers of implants/dam, live foetuses/litter, or resorptions/litter were not significantly altered by exposures. No significant effect on foetal sex ratio was seen. Mean weights and lengths of the offspring of rabbits exposed to sulphuric acid were not significantly different from their respective control values. The offspring of rabbits exposed to sulphuric acid did not have a significantly higher incidence of malformations than controls. In rabbits, an increased incidence of small non-ossified areas in the skull bones (a minor variation) was the only statistically significant evidence of embryotoxicity observed among the offspring of rabbits exposed to 20 mg/m3.
Inhalation of 5 or 20 mg sulphuric acid/m3 did cause any signs of toxicity. The incidence of pregnancy was not significantly altered among the mice. Bodyweight gain of exposed mice was not significantly affected but a significant decrease in the amount of food consumed during the first few days of exposure was observed at 20 mg/m3. The liver weight of dams was significantly decreased in mice exposed to 20 mg/m3 compared to controls. No evidence of toxicity that could be attributed to sulphuric acid was observed in the respiratory tract of mice (nasal turbinates, trachea and lungs). The mean numbers of implants/dam, live foetuses/litter, or resorptions/litter were not significantly altered by exposures. No significant effect on foetal sex ratio was seen. Mean weights and lengths of the offspring of mice exposed to sulphuric acid were not significantly different from their respective control values. The offspring of mice exposed to sulphuric acid did not have a significantly higher incidence of malformations than controls. Inhalation of sulphuric acid did not increase the incidence of minor skeletal variants among the offspring of exposed mice.
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
- 19.3 mg/m³
- Study duration:
- subchronic
- Experimental exposure time per week (hours/week):
- 49
- Species:
- other: rabbit and mouse
- Quality of whole database:
- The published study, comparable to OECD 414, is considered reliable and adequate for characterising the developmental toxicity of the substance
Effect on developmental toxicity: via dermal route
- Endpoint conclusion:
- no study available
Additional information
In the study by Murray, F..J. et al (1979), mated female mice were exposed (whole body) to atmospheres containing aerosols of sulphuric acid at 0, 5.7 mg/m3 (MMAD 1.6 um) or 19.3 mg/m3 (MMAD 2.4 um) on Days 6 -15 of gestation (mice). Animals were sacrificed at Days 18 (mice). Evidence of mild toxicity was seen in maternal mice (reduced food consumption) at 20 mg/m3. No evidence of teratogenicity or developmental toxicity was seen. Inseminated female rabbits were exposed (whole body) to atmospheres containing aerosols of sulphuric acid at 0, 5.7 mg/m3 (MMAD 1.6 um) or 19.3 mg/m3 (MMAD 2.4 um) on Days 6 -18 of gestation (rabbits). Animals were sacrificed at Day 29 of gestation(rabbits). Evidence of mild toxicity was seen in maternal rabbits (reduced weight gain, respiratory tract effects) at 20 mg/m3. No evidence of teratogenicity or developmental toxicity was seen.
No evidence of teratogenicity or developmental toxicity was seen in the mouse and rabbit in an inhalation study comparable to OECD 414, and at exposure levels (19.3 mg/m3) sufficient to cause mild maternal toxicity. Given the results of this study and the dataset as a whole, which indicates the absence of systemic exposure, further testing (e.g. in the rat) is not proposed.
It is calculated that exposure to sulphuric acid at the TWA will result in negligible additional systemic sulphate exposure, which will not have any effect on foetal development.
Testing cannot be justified on scientific grounds (given the absence of
systemic exposure, detailed above) or (due to the corrosive nature of
the substance) for reasons of animal welfare. Both the OECD SIDS (2001)
and the US ATSDR (1998) concluded that the exiting information do not
show any evidence of developmental toxicity and that further testing is
not required based on a lack of systemic exposure.
Toxicity to reproduction: other studies
Additional information
No studies have been performed and none are proposed in the absence of any effects in the existing dataset and given the absence of systemic exposure. Further studies cannot be justified on scientific grounds and (given the corrosive nature of the substance) also cannot be justified for reasons of animal welfare.
Justification for classification or non-classification
No classification is proposed for reproductive or developmental toxicity. The existing data and the absence of systemic exposure do not indicate that classification is required.
Additional information
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.
![ECHA](/o/diss-blank-theme/images/factsheets/A-REACH/factsheet/echa_logo.png)