Asphalt, sulfonated, sodium salt

Toxicity to reproduction

Administrative data

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2005-08-08 to 2007-02-13

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Cross-reference 2

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2007

Report date:

2007

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)

Deviations:

no

Principles of method if other than guideline:

N/A

GLP compliance:

yes (incl. QA statement)

Remarks:

Certificate from "The Department of Health of the Government of the United Kingdom".

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Asphalt, sulfonated, sodium salt

EC Number:

269-212-0

EC Name:

Asphalt, sulfonated, sodium salt

Cas Number:

68201-32-1

Molecular formula:

UVCB

IUPAC Name:

Asphalt, sulfonated, sodium salt

Details on test material:

- Name of test material (as cited in study report): Sodium Sulfonated Asphalt
- Molecular formula (if other than submission substance): N/A
- Molecular weight (if other than submission substance): N/A
- Smiles notation (if other than submission substance): N/A
- InChl (if other than submission substance): N/A
- Structural formula attached as image file (if other than submission substance): N/A
- Substance type: active
- Physical state: black solid

Test animals

Species:

rat

Strain:

other: Sprague-Dawley Crl:CD (SD) IGS BR

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River (UK) Limited, Margate, Kent
- Age at study initiation: (P) 8 weeks
- Weight at study initiation: (P) Males: 278-333 g; Females: 169-228 g
- Fasting period before study: N/A
- Housing: Initially, all animals were housed in groups of five in polypropylene cages with stainless steel grid floors and tops, suspended over polypropylene trays lined with absorbent paper. During the mating phase, animals were transferred to similar cages on a one male: one female basis. Following evidence of successful mating, the males were returned to their original cages. Mated females were housed individually during gestation and lactation, in polypropylene cages with solid floors and stainless steel lids, furnished with softwood flakes.
- Use of restrainers for preventing ingestion (if dermal): yes/no
- Diet (e.g. ad libitum): ad libitum (Rodent PMI 5002 (Certified) diet, BCM IPS limited, London, UK)
- Water (e.g. ad libitum): ad libitum (drinking water supplied from polycarbonate bottles attached to the cage)
- Acclimation period: The animals were acclimatized for 10 days, during which time their health status was assessed.
- Other: On receipt, the animals were examined for signs of ill-health or injury.


ENVIRONMENTAL CONDITIONS
- Temperature (deg. C): Temperature controls were set to achieve target values of 21+/- 2 deg C. Deviations from this target were considered not to affect the purpose or integrity of the study.
- Humidity (%): Relative humidity controls were set to achieve target values of 55+/-15 %. Deviations from this target were considered not to affect the purpose or integrity of the study.
- Air changes (per hr): at least 15
- Photoperiod (hrs dark / hrs light): 12/12
-Other: Environmental conditions were continuously monitored by a computerized system, and print-outs of hourly mean temperatures and humidities were included in the study records.


IN-LIFE DATES: From: N/A To: N/A

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: distilled water

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: Formulations were prepared weekly and stored at approximately +4 deg C in the dark.

DIET PREPARATION
- Rate of preparation of diet (frequency): N/A
- Mixing appropriate amounts with (Type of food): N/A
- Storage temperature of food: N/A


VEHICLE
- Justification for use and choice of vehicle (if other than water): N/A
- Concentration in vehicle: 0, 50, 100 and 200 mg/mL
- Amount of vehicle (if gavage): 5 mL/kg (The volume administered to each animal was based on the most recent body weight and was adjusted weekly for males, recovery animals and females during maturation and on Days 0, 7, 14 and 20 of gestation, and on Days 1 and 4 of lactation).
- Lot/batch no. (if required): N/A
- Purity: N/A

Details on mating procedure:

- M/F ratio per cage: On Day 15, all animals (excluding Recovery animals) were paired on a 1 male:1 female basis within each dose group.
- Length of cohabitation: until evidence of mating (maximum of 14 days)
- Proof of pregnancy: Cage tray-liners were checked each morning for the presence of ejected copulation pugs, and each female was examined for the presence of a copulation plug in the vagina. A vaginal smear was prepared for each female and the stage of the oestrous cycle or the presence of sperm was recorded. The presence of sperm within the vaginal smear and/or vaginal plug in situ was taken as positive evidence of mating. Sperm in smear referred to as day 0 of gestation.
- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility: N/A
- Further matings after two unsuccessful attempts: N/A
- After successful mating each pregnant female was caged (how): Mated females were housed individually during gestation and lactation, in polypropylene cages with solid floors and stainless steel lids, furnished with softwood flakes.
- Any other deviations from standard protocol: N/A

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The concentration of the test substance in the formulations was determined spectrophotometrically. The test substance formulations were sampled and analyzed within three days of preparation.
The test substance formulations were diluted with DMSO to give a final, theoretical test substance concentration of approximately 0.025 mg/mL.
Standard solutions of test substance were prepared in DMSO at a nominal concentration of 0.025 mg/mL.
The standard and sample solutions were analyzed spectrophotometrically using the following conditions:
Spectrophotometer: Perkin-Elmer Lambda 20 and Camspec M330
Wavelength: ~268 nm
Cell path length: 10 mm
Reference medium: DMSO
For homogeneity determination, test substance formulations were mixed thoroughly and samples were taken from the top, middle and bottom of the container, shaking between sampling. Sampling was performed in triplicate.
For stability determinations, the test substance formulations were sampled and analyzed initially and then after storage at approximately +4degrees C in the dark for eighteen days.
Results showed the formulations to be stable for at least fourteen days, and the formulations were within acceptable limits of the nominal concentration.

Duration of treatment / exposure:

up to 54 consecutive days (males were killed on Day 43 and females were killed on Day 5 post partum)

Frequency of treatment:

The test substance (or vehicle alone for control animals) was administered daily (except for females during littering/parturition where applicable) by gavage using a stainless steel cannula attached to a disposable plastic syringe.

Details on study schedule:

- F1 parental animals not mated until [...] weeks after selected from the F1 litters: N/A
- Selection of parents from F1 generation when pups were [...] days of age: N/A
- Age at mating of the mated animals in the study: [...] weeks: N/A
-Other: Each pregnant female was observed at approximately 0830, 1230, and 1630 hours and around the period of expected parturition. Observations were carried out at approximately 0830 and 1230 hours on weekends and public holidays. The following was recorded for each female:
1. date of mating
2. date and time of observed start of parturition
3. date and time of observed completion of parturition
4. duration of gestation

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

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

Doses / concentrations 2

Dose / conc.:

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

Doses / concentrations 3

Dose / conc.:

1 000 mg/kg bw/day (actual dose received)

No. of animals per sex per dose:

10 animals/sex/dose

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: The dose levels were chosen based on the results of a preliminary range-finder.
- Rationale for animal assignment (if not random): The animals were allocated to dose groups using a randomization procedure based on stratified body weights, and the group mean body weights were then determined to ensure similarity between the dose groups.
- Other: N/A

Positive control:

No

Examinations

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: see below
- Cage side observations included: All animals were examined for overt signs of toxicity, ill-health and behavioral change immediately before and after dosing, and one and five hours after dosing, during the working week. Animals were observed immediately before and after dosing, and one hour after dosing on weekends (except for females during parturition where applicable). During the treatment-free period, animals were observed twice daily, morning and afternoon (once daily at weekends). All observations were recorded.


DETAILED CLINICAL OBSERVATIONS: Neurobehavioral tests were performed. See below for details.
- Time schedule: N/A


BODY WEIGHT: Yes
- Time schedule for examinations: Individual body weights were recorded on day 1 (prior to dosing) and then weekly for males until termination. Females were weighed weekly until mating was evident. Body weights were then recorded on Days 0, 7, 14 and 20 post coitum, and on Days 1 and 4 post partum. Recovery animals were weighed on Day 1, and then weekly until termination.


FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption calculated: Yes; During the maturation period, weekly food consumption was recorded for each cage of adults. This was continued for males after the mating phase. For females showing evidence of mating, food consumption was recorded for the periods covering Days 0-7, 7-14 and 14-24. For females with live litters, food consumption was recorded on Days 1 and 4 post partum. Food consumption for recovery animals was recorded weekly until termination. Food consumption was reported as g/rat/day. Food efficiency was also determined by dividing the group mean body weight gain (g/rat) by the group mean food consumption (g/rat/day) times number of days.
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No


WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
- Time schedule for examinations: Water intake was observed daily by visual inspection of water bottles for any overt change.


OTHER:
1. Neurobehavioral: Prior to the start of treatment and at weekly intervals thereafter, all non-recovery animals were observed for signs of functional/behavioral toxicity. Functional performance tests were also performed on five selected males and females per dose level, prior to termination, together with assessment of sensory reactivity to various stimuli.
Detailed individual clinical observations were performed for each non-recovery animal using a purpose-built arena. The following behavioral assessment parameters were observed: gait, tremors, twitches, convulsions, bizarre/abnormal/stereotypic behavior, salivation, pilo-erection, exophthalmia, lachrymation, hyper/hypothermia, skin color, respiration, palpebral closure, urination, defecation, transfer arousal, and tail elevation.
Purpose-built 44 infra-red beam automated activity monitors were used to assess motor activity. Animals were randomly allocated to the activity monitors. The tests were performed at approximately the same time each day, under similar laboratory condition. The evaluation period was thirty minutes for each animal. The percentage of time each animal was active and mobile was recorded for the overall thirty minute period and also during the final 20% of the period (considered to be the asymptotic period).
For forelimb/hindlimb grip strength, an automated grip strength meter was used. Each animal was allowed to grip the proximal metal bar of the meter with its forepaws. The animal was pulled by the base of the tail until its grip was broken. The animal was drawn along the trough of the meter by the tail until its hind paws gripped the distal metal bar. The animal was pulled by the base of the tail until its grip was broken. A record of the force required to break the grip for each animal was made. Three consecutive trials were performed for each animal.
Each animal was individually assessed for sensory reactivity to auditory, visual and proprioceptive stimuli. The following parameters were observed: grasp response, vocalization, toe pinch, tail pinch, finger approach, touch escape, pupil reflex, startle reflex and blink reflex.

2. Haematological and blood chemical investigations were performed on five non-recovery males and five non-recovery females randomly selected from each test and control group on Day 14 (day prior to pairing). Blood chemical and haematological assessments were performed on five non-recovery animals at termination and on Recovery animals on Day 57. Blood samples were obtained from the lateral tail vein. Animals were not fasted prior to sampling. For haematology, The following parameters were measured on blood collected into tubes containing potassium EDTA anti-coagulant: haemoglobin, erythrocyte count, haematocrit, erythrocyte indices (mean corpuscular haemoglobin, mean corpuscular volume and mean corpuscular haemoglobin concentration), total leucocyte count, differential leucocyte count (neutrophils, lymphocytes, monocytes, eosinophils, basophils), platelet count, reticulocyte count (cresyl blue stained slides were prepared, but reticulocytes were not assessed), prothrombin time (assessed by Thrombomax HS with Calcium) and activated partial thromboplastin time (assessed by Actin FS using samples collected into sodium citrate solution). For Blood chemistry, the following parameters were measured on plasma from blood collected into tubes containing lithium heparin anti-coagulant: urea, glucose, total protein, albumin, albumin/globulin ratio, sodium, potassium chloride, calcium, inorganic phosphorus, aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, creatinine, total cholesterol, and total bilirubin.


3. The following parameters were measured on collected urine: volume, specific gravity, pH, protein, glucose, ketones, bilirubin, urobilinogen, reducing substances, blood.

Oestrous cyclicity (parental animals):

During mating, a vaginal smear was prepared daily for each female, and the stage of the oestrous cycle was recorded.

Sperm parameters (parental animals):

Parameters examined in male parental generations: testis weight and epididymides weight

Litter observations:

STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: N/A
- If yes, maximum of [...] pups/litter ([...]/sex/litter as nearly as possible: N/A

PARAMETERS EXAMINED
The following parameters were examined in offspring: On completion of parturition, the number of live and dead offspring was recorded. For each litter the following was recorded: number of pups born, number and sex of pups alive recorded daily and reported on Days 1 and 4 post partum, clinical condition of pups from birth to Day 4 post partum, and individual pup and litter weights on Days 1 and 4 post partum. All offspring were observed for the detachment of pinna and assessed for reflexological response to stimuli by assessing surface righting reflex on Day 1 post partum.

GROSS EXAMINATION OF DEAD PUPS:
Yes; all animals (both those that died and ones that were sacrificed) were subjected to a full external and internal examination, and any macroscopic abnormalities were recorded.

Postmortem examinations (parental animals):

SACRIFICE
- Male animals: Males were killed by intravenous overdose of sodium pentobarbitone followed by exsanguinations on Day 43.
- Maternal animals: Females were killed by intravenous overdose of sodium pentobarbitone followed by exsanguinations on Day 5 post partum.


GROSS NECROPSY
- Gross necropsy consisted of a full external and internal examination and any macroscopic abnormalities were recorded. In addition, corpora lutea of all ovaries from pregnant females were counted at necropsy. Uterine implantation sites were counted also.

HISTOPATHOLOGY / ORGAN WEIGHTS
The following organs, removed from the five selected non-recovery and recovery adult animals that were killed at the end of the study, were dissected free from fat and weighed before fixation: adrenals; brain; epididymides*; heart; kidneys; liver; ovaries*; spleen; testes*; thymus (*=organs weighed from all animals).
Samples of the following tissues were preserved from five males and five females from each doses group: adrenals; aorta (thoracic); bone and bone marrow (femur including stifle joint; sternum); brain (including cerebrum, cerebellum and pons); caecum; coagulating gland*; colon; duodenum; epididymides*; eyes; gross lesions; heart; ileum; jejunum; kidneys; liver; lungs (with bronchi); lymph nodes(cervical and mesenteric); mammary gland; muscle (skeletal); ovaries*; pancreas; pituitary*; prostate*; oesophagus; rectum; salivary glands (submaxillary); sciatic nerve; seminal vesicles*; skin (hind limb); spinal cord (cervical); spleen; stomach; thyroid/parathyroid; trachea; testes*; thymus; urinary bladder; uterus/cervix*; vagina* (*=tissues also removed from the remaining animals).
The tissues from five selected control and high dose animals were prepared as paraffin blocks, sectioned at nominal thickness of 5 mm and stained with haematoxylin and eosin for subsequent microscopic examination. The tissues with (*) next to them from the remaining control and high dose animal were also processed.

Postmortem examinations (offspring):

SACRIFICE
- The offspring were sacrificed at 5 days of age.
- These animals were subjected to a full external and internal examination, and any macroscopic abnormalities were recorded.


GROSS NECROPSY
- Gross necropsy consisted of a full external and internal examination, and any macroscopic abnormalities were recorded.




HISTOPATHOLOGY / ORGAN WEIGHTS
The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively: N/A

Statistics:

Statistical Methods: Haematological, blood chemical, organ weight (absolute and relative to terminal bodyweight) weekly bodyweight gain, litter weights, offspring bodyweights and quantitative functional performance data were assessed for dose response relationships by linear regression analysis followed by one way analysis of variance (ANOVA) incorporating Leven's test for homogeneity of variance. Where variances were shown to be homogeneous, pairwise comparisons were conducted using Dunnett's test. Where Leven's test showed unequal variances, the data were analysed using non-parametric methods: Kruskal-Wallis ANOVA and Mann-Whitney 'U' test. The non-parametric methods were also used to analyse implantation loss, offspring sex ratio and landmark developmental markers.
The haematology variable basophils was not analysed since consistently greater than 30% of the data were recorded as the same value.

Reproductive indices:

see below

Offspring viability indices:

see below

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

no effects observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Other effects:

not specified

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

not specified

Reproductive function: sperm measures:

not specified

Reproductive performance:

no effects observed

Details on results (P0)

CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS): No unscheduled deaths during the study. No clinically observable signs of toxicity were detected in animals of either sex treated with 250, 500 or 1000 mg/kg bw/day. Episodes of respiratory pattern changes and generalised fur staining were evident throughout the treatment groups during the treatment period.


BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS): There were no adverse effects on bodyweights for males through the study or for females during the two week maturation period. Recovery males treated with 1000 mg/kg showed a statistically significant increase in body weight gain during week 6 and a statistically significant reduction in body weight gain during week 7. In the absence of similar effects seen in non-recovery males, the intergroup differences were considered of no toxicological significance. There were no adverse effects on bodyweight gains for females during the gestation or lactation phase of the study. No adverse effects on food consumption or food efficiency were detected for males throughout the treatment period or for females throughout the two week maturation period. There were no adverse effects on food consumption for females during the gestation or lactation phase of the study.

TEST SUBSTANCE INTAKE (PARENTAL ANIMALS): N/A


REPRODUCTIVE FUNCTION: ESTROUS CYCLE (PARENTAL ANIMALS): N/A


REPRODUCTIVE FUNCTION: SPERM MEASURES (PARENTAL ANIMALS): N/A


REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS): There were no treatment-related effects on mating performance or fertility. The distribution of pre-coital intervals for treated animals was comparable to controls with the majority of animals showing positive evidence of mating within four days of pairing. The distribution of animals with precoital intervals in excess of four days showed no treatment-related trends. Only one control female failed to achieve pregnancy. There were no significant intergroup differences in gestation length or parturition. The distribution for treated females was comparable to controls. In total there were 9, 10, 10, and 10 females at 0, 250, 500 and 1000 mg/kg, respectively, who gave birth to a live litter and successfully reared young to Day 5 of age. The mean numbers of corpora lutea observed for treated females were considered to be good and did not indicate any adverse effect of treatment.


ORGAN WEIGHTS (PARENTAL ANIMALS): No treatment-related effects were detected in the organ weights measured. Statistical analysis of the data revealed no significant intergroup differences.

GROSS PATHOLOGY (PARENTAL ANIMALS): No treatment-related macroscopic abnormalities were detected at terminal kill.


HISTOPATHOLOGY (PARENTAL ANIMALS): No treatment-related microscopic changes were detected.


OTHER FINDINGS (PARENTAL ANIMALS)
-Behavioural Assessments: All inter- and intra-group differences in urination, defecation and transfer arousal scores were considered to be a result of normal variation for rats of the strain and age used and were of no toxicological importance.
- Functional Performance Tests: No treatment-related effects were detected. Statistical analysis of the data revealed no significant intergroup differences.
- Sensory Reactivity Assessments: No treatment-related effects were detected. All inter- and intra-group differences in sensory reactivity scores were considered to be a result of normal variation for rats of the strain and ages used and were of no toxicological importance.
-Water Consumption: No intergroup differences were detected.
- Haematology: No treatment-related effects were detected. Males treated with 1000 mg/kg/day showed statistically significant reductions in clotting time at the terminal bleed. In the absence of any supporting data to suggest an effect of treatment, this intergroup difference was considered to be of no toxicological significance.
- Blood Chemistry: No treatment-related effects were detected. Males treated with 1000 mg/kg/day showed a statistically significant reduction in plasma calcium concentration on Day 14. The effect continued at Day 43 and extended to males from the remaining treatment groups. Males treated with 1000 and 250 mg/kg/day also showed a statistically significant reduction in plasma inorganic phosphorus at Day 14. In the absence of a dose related response, or in the absence of any supporting data to suggest an effect of treatment, these intergroup differences were considered to be of no toxicological significance. Males treated with 1000 mg/kg/day showed a statistically significant reduction in plasma urea. In the absence of any histopathological correlates, or a dose-related response, the intergroup difference was considered of no toxicological importance. Recovery males treated with 1000 mg/kg/day showed a statistically significant increase in plasma creatinine levels together with a reduction in plasma inorganic phosphorus. Recovery females treated with 1000 mg/kg/day showed a statistically significant reduction in plasma albumin and albumin/globulin ratio, together with an increase in plasma inorganic phosphorus. In the absence of similar treatment-related effects for non-recovery animals at the end of the dosing period, the intergroup differences were considered to be incidental and of no toxicological importance.
- Urinalysis: No treatment-related effects were detected.

Effect levels (P0)

Key result

Dose descriptor:

NOEL

Effect level:

>= 1 000 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No toxicologically significant changes in the parameters measured (general or reproductive).

Target system / organ toxicity (P0)

Critical effects observed:

no

Results: F1 generation

General toxicity (F1)

Clinical signs:

no effects observed

Mortality / viability:

no mortality observed

Body weight and weight changes:

no effects observed

Sexual maturation:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

no effects observed

Histopathological findings:

not specified

Details on results (F1)

VIABILITY (OFFSPRING): Prenatal losses and resultant litter size at Day 1 for treated animals were similar to controls. Postnatal survival was unaffected in all treated groups, with litter size at Day 4 again being similar to controls.


CLINICAL SIGNS (OFFSPRING): No toxicologically significant clinical findings were observed. The type and incidence of clinical observations recorded for offspring throughout the dose groups were consistent with what is normally expected of the age examined and were of no toxicological importance.


BODY WEIGHT (OFFSPRING): Mean litter weights, mean offspring bodyweights and mean bodyweight gain of the offspring were considered to have been unaffected by maternal exposure in females treated with 250, 500 or 1000 mg/kg/day.


SEXUAL MATURATION (OFFSPRING): N/A


ORGAN WEIGHTS (OFFSPRING): N/A


GROSS PATHOLOGY (OFFSPRING): The macroscopic findings observed for interim and terminal kill offspring throughout the dose groups were consistent with normally expected low incidence findings in pups of the age examined and were of no toxicological importance.



HISTOPATHOLOGY (OFFSPRING): N/A


OTHER FINDINGS (OFFSPRING): No treatment-related effects on offspring growth or development were detected. Intergroup differences in offspring maturation (as assessed by pinna unfolding) and reflexological assessment (percentage successful at surface righting) did not indicate any adverse effects of treatment at 250, 500 or 1000 mg/kg/day. Statistical analysis of the data revealed no significant intergroup differences.

Effect levels (F1)

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 effect of treatment was detected on offspring development.

Target system / organ toxicity (F1)

Critical effects observed:

no

Overall reproductive toxicity

Reproductive effects observed:

no

Any other information on results incl. tables

N/A

Applicant's summary and conclusion

Conclusions:

The oral administration of sodium sulfonated asphalt to rats by gavage at dose levels of 1000, 500 or 250 mg/kg/day produced no toxicologically significant changes in the parameters measured. The No Observed Effect Level (NOEL) for parental and offspring toxicity was therefore considered to be ≥1000 mg/kg/day.

No effect of treatment was detected on reproduction or offspring development and the No Observed Effect Level (NOEL) for reproductive and developmental toxicity was also considered to be 1000 mg/kg/day.

Executive summary:

N/A