Phosphonic acid, calcium salt (2:1)

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Remarks:

combined repeated dose and reproduction / developmental screening

Type of information:

experimental study

Adequacy of study:

key study

Study period:

January-May 2013

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

Reference

Endpoint:

one-generation reproductive toxicity

Remarks:

based on generations indicated in Effect levels (migrated information)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2013, January-May

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

Animal supply and acclimatisationA total of 90 Sprague Dawley rats (45 males and 45 virgin females), 6 to 7 weeks old and weighing 176 to 200 g for males and 151 to 175 g for females, have been ordered from Charles River Italia S.p.A., Calco (Lecco), Italy.After arrival the weight range for each sex have been determined and the animals have been temporarily identified within the cage by means of a coloured mark on the tail. A health check has then been performed by a veterinarian.An acclimatisation period of approximately 2 weeks has been allowed before the start of treatment, during which time the health status of the animals has been assessed by thorough observations. Rats considered unsatisfactory have been killed and were appropriate subjected to pathological examination.Animal husbandryThe animals have been housed in a limited access rodent facility. Animal room controls have been set to maintain temperature and relative humidity at 22°C +/- 2°C and 55%+/- 15% respectively; actual conditions have been monitored, recorded and the records retained. There will be approximately 15 to 20 air changes per hour and the rooms will be lit by artificial light for 12 hours each day.From arrival to pairing, animals have been housed up to 5 of one sex to a cage, in polisulphone solid bottomed cages measuring 59.5x38x20 cm (Techniplast Gazzada S.a.r.l., Buguggiate, Varese). Nesting material has been provided inside suitable bedding bags and changed at least twice a week.During mating, animals have been housed one male to one female in clear polycarbonate cages measuring approximately 43x27x18cm with a stainless steel mesh lid and floor (Techniplast – Gazzada S.a.r.l.). Each cage tray holded absorbent material which has been inspected and changed daily.After mating, the males have been recaged as they were before mating, the females have been transferred to individual solid bottomed cages (Techniplast Gazzada S.a.r.l.) for the gestation period, birth and lactation. Suitable nesting material has been provided and has been changed as necessary.Drinking water has been supplied ad libitum to each cage via water bottles, except in the case of urinalysis investigations.A commercially available laboratory rodent diet (4 RF 21, Mucedola S.r.l., Via G. Galilei, 4, 20019, Settimo Milanese (MI), Italy) will be offered ad libitum throughout the study.There is no information available to indicate that any non-nutrient substance likely to influence the effect of the test item is present in the drinking water or the diet. Records of analyses of water and diet are kept on file at RTC.Dated and signed records of activities relating to the day to day running and maintenance of the study in the animal house will be recorded.

Route of administration:

oral: gavage

Vehicle:

water

Analytical verification of doses or concentrations:

yes

Duration of treatment / exposure:

MalesAnimals have been dosed once a day, 7 days a week, for a minimum of 2 consecutive weeks prior to pairing and thereafter through the day before necropsy.Dose volumes will be adjusted once per week for each animal according to the last recorded body weight.FemalesAnimals will be dosed once a day, 7 days a week, for a minimum of 2 consecutive weeks prior to pairing and thereafter during pairing , post coitum and post partum periods until Day 3 post partum or the day before sacrifice. Dose volumes will be adjusted once per week for each animal according to the last recorded body weight.During the gestation period, dose volumes will be calculated according to individual body weight on Days 0, 7, 14 and 20 post coitum and on Day 1 post partum. Thereafter individual dose volumes will remain constant.

Remarks:

Doses / Concentrations:10 mg/Kg bwBasis:actual ingested

Remarks:

Doses / Concentrations:100 mg/Kg bwBasis:actual ingested

Remarks:

Doses / Concentrations:1000 mg/Kg bwBasis:actual ingested

No. of animals per sex per dose:

10 animal per sex per dose

Control animals:

yes, concurrent vehicle

Details on study design:

The test item have been administered orally by gavage at a dose volume of 10 mL/kg body weight. Control animals received the vehicle alone at the same dose volume.The dose has been administered to each animal on the basis of the most recently recorded body weight and the volume administered has been recorded for each animal

Parental animals: Observations and examinations:

MortalityEarly in each working day in the morning and in the afternoon. At weekends and Public Holidays a similar procedure will be followed except that the final check has been carried out at approximately mid-day.Clinical signsOnce before commencement of treatment and at least once daily during the study,Clinical observations (Functional Observation Battery Tests)Once before commencement of treatment and at least once a week thereafterGrip strength and sensory reactivity to stimuliOnce during the study, towards the end of treatment, on 5 males and 5 females randomly selected from each groupMotor activity assessment (MA)Once during the study, towards the end of treatment, on 5 males and 5 females randomly selected from each groupFood consumptionRecorded weekly (whenever possible) during the pre-mating period starting from allocation. Individual food consumption for the females have been measured on gestation Days 7, 14 and 20 starting from Day 0 post coitum and on Day 4 post partum starting from Day 1 post partum.Body weightMales weekly from allocation to termination.Females weekly from allocation to positive identification of mating and on gestation Days 0, 7, 14 and 20.Dams Days 1 and 4 post partum.

Oestrous cyclicity (parental animals):

Vaginal smearsVaginal smears has been taken daily in the morning starting two weeks before pairing until a positive identification of copulation is made.

Litter observations:

Pups identification, weight and observationAs soon as possible, after parturition is considered complete (Day 0 or 1 post partum), .Once daily for all litters

Clinical signs:

no effects observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Other effects:

no effects observed

Reproductive function: oestrous cycle:

no effects observed

Reproductive function: sperm measures:

no effects observed

Reproductive performance:

no effects observed

Dose descriptor:

NOEL

Effect level:

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

Based on:

test mat.

Sex:

male/female

Basis for effect level:

clinical signs

body weight and weight gain

food consumption and compound intake

organ weights and organ / body weight ratios

gross pathology

histopathology: non-neoplastic

reproductive function (oestrous cycle)

reproductive function (sperm measures)

reproductive performance

Clinical signs:

no effects observed

Mortality / viability:

no mortality observed

Body weight and weight changes:

no effects observed

Sexual maturation:

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings:

no effects observed

Dose descriptor:

NOEL

Generation:

F1

Effect level:

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

Based on:

test mat.

Sex:

male/female

Basis for effect level:

sexual maturation

clinical signs

mortality

body weight and weight gain

organ weights and organ / body weight ratios

gross pathology

histopathology: non-neoplastic

Reproductive effects observed:

not specified

Conclusions:

NOEL oral rat ≥ 1000 mg/kg bw/day

Cross-reference 2

Reason / purpose for cross-reference:

reference to same study

Reference

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2013, January-May

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

other: OECD 422

GLP compliance:

yes (incl. QA statement)

Species:

rat

Strain:

Sprague-Dawley

Details on test animals or test system and environmental conditions:

Animal supply and acclimatisationA total of 90 Sprague Dawley rats (45 males and 45 virgin females), 6 to 7 weeks old and weighing 176 to 200 g for males and 151 to 175 g for females, have been ordered from Charles River Italia S.p.A., Calco (Lecco), Italy.After arrival the weight range for each sex have been determined and the animals have been temporarily identified within the cage by means of a coloured mark on the tail. A health check has then been performed by a veterinarian.An acclimatisation period of approximately 2 weeks has been allowed before the start of treatment, during which time the health status of the animals has been assessed by thorough observations. Rats considered unsatisfactory have been killed and were appropriate subjected to pathological examination.Animal husbandryThe animals have been housed in a limited access rodent facility. Animal room controls have been set to maintain temperature and relative humidity at 22°C +/- 2°C and 55%+/- 15% respectively; actual conditions have been monitored, recorded and the records retained. There will be approximately 15 to 20 air changes per hour and the rooms will be lit by artificial light for 12 hours each day.From arrival to pairing, animals have been housed up to 5 of one sex to a cage, in polisulphone solid bottomed cages measuring 59.5x38x20 cm (Techniplast Gazzada S.a.r.l., Buguggiate, Varese). Nesting material has been provided inside suitable bedding bags and changed at least twice a week.During mating, animals have been housed one male to one female in clear polycarbonate cages measuring approximately 43x27x18cm with a stainless steel mesh lid and floor (Techniplast – Gazzada S.a.r.l.). Each cage tray holded absorbent material which has been inspected and changed daily.After mating, the males have been recaged as they were before mating, the females have been transferred to individual solid bottomed cages (Techniplast Gazzada S.a.r.l.) for the gestation period, birth and lactation. Suitable nesting material has been provided and has been changed as necessary.Drinking water has been supplied ad libitum to each cage via water bottles, except in the case of urinalysis investigations.A commercially available laboratory rodent diet (4 RF 21, Mucedola S.r.l., Via G. Galilei, 4, 20019, Settimo Milanese (MI), Italy) will be offered ad libitum throughout the study.There is no information available to indicate that any non-nutrient substance likely to influence the effect of the test item is present in the drinking water or the diet. Records of analyses of water and diet are kept on file at RTC.Dated and signed records of activities relating to the day to day running and maintenance of the study in the animal house will be recorded.

Route of administration:

oral: gavage

Vehicle:

water

Analytical verification of doses or concentrations:

yes

Duration of treatment / exposure:

MalesAnimals have been dosed once a day, 7 days a week, for a minimum of 2 consecutive weeks prior to pairing and thereafter through the day before necropsy.Dose volumes will be adjusted once per week for each animal according to the last recorded body weight.FemalesAnimals will be dosed once a day, 7 days a week, for a minimum of 2 consecutive weeks prior to pairing and thereafter during pairing , post coitum and post partum periods until Day 3 post partum or the day before sacrifice. Dose volumes will be adjusted once per week for each animal according to the last recorded body weight.During the gestation period, dose volumes will be calculated according to individual body weight on Days 0, 7, 14 and 20 post coitum and on Day 1 post partum. Thereafter individual dose volumes will remain constant.

Remarks:

Doses / Concentrations:10 mg/Kg bw Basis:actual ingested

Remarks:

Doses / Concentrations:100 mg/Kg bw Basis:actual ingested

Remarks:

Doses / Concentrations:1000 mg/Kg bw Basis:actual ingested

No. of animals per sex per dose:

10 per sex per dose

Control animals:

yes, concurrent vehicle

Details on maternal toxic effects:

Maternal toxic effects:no effects

Dose descriptor:

NOEL

Effect level:

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

Based on:

test mat.

Basis for effect level:

other: developmental toxicity

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Dose descriptor:

NOEL

Effect level:

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

Based on:

test mat.

Basis for effect level:

other: teratogenicity

Abnormalities:

not specified

Developmental effects observed:

not specified

Conclusions:

NOEL oral rat ≥ 1000 mg/kg bw/day

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

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)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Potassium Phosphonate KH2PO3/K2HPO3

IUPAC Name:

Potassium Phosphonate KH2PO3/K2HPO3

Details on test material:

Batch number : 20111216-014Date of expiry : 13 December 2013Appearance : White clear fluid solution Storage conditions : Room temperature

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

Animal supply and acclimatisationA total of 90 Sprague Dawley rats (45 males and 45 virgin females), 6 to 7 weeks old and weighing 176 to 200 g for males and 151 to 175 g for females, have been ordered from Charles River Italia S.p.A., Calco (Lecco), Italy.After arrival the weight range for each sex have been determined and the animals have been temporarily identified within the cage by means of a coloured mark on the tail. A health check has then been performed by a veterinarian.An acclimatisation period of approximately 2 weeks has been allowed before the start of treatment, during which time the health status of the animals has been assessed by thorough observations. Rats considered unsatisfactory have been killed and were appropriate subjected to pathological examination. Animal husbandry The animals have been housed in a limited access rodent facility. Animal room controls have been set to maintain temperature and relative humidity at 22°C +/- 2°C and 55%+/- 15% respectively; actual conditions have been monitored, recorded and the records retained. There will be approximately 15 to 20 air changes per hour and the rooms will be lit by artificial light for 12 hours each day. From arrival to pairing, animals have been housed up to 5 of one sex to a cage, in polisulphone solid bottomed cages measuring 59.5x38x20 cm (Techniplast Gazzada S.a.r.l., Buguggiate, Varese). Nesting material has been provided inside suitable bedding bags and changed at least twice a week. During mating, animals have been housed one male to one female in clear polycarbonate cages measuring approximately 43x27x18cm with a stainless steel mesh lid and floor (Techniplast – Gazzada S.a.r.l.). Each cage tray holded absorbent material which has been inspected and changed daily.After mating, the males have been recaged as they were before mating, the females have been transferred to individual solid bottomed cages (Techniplast Gazzada S.a.r.l.) for the gestation period, birth and lactation. Suitable nesting material has been provided and has been changed as necessary.Drinking water has been supplied ad libitum to each cage via water bottles, except in the case of urinalysis investigations.A commercially available laboratory rodent diet (4 RF 21, Mucedola S.r.l., Via G. Galilei, 4, 20019, Settimo Milanese (MI), Italy) will be offered ad libitum throughout the study.There is no information available to indicate that any non-nutrient substance likely to influence the effect of the test item is present in the drinking water or the diet. Records of analyses of water and diet are kept on file at RTC. Dated and signed records of activities relating to the day to day running and maintenance of the study in the animal house have been recorded.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Analytical verification of doses or concentrations:

yes

Duration of treatment / exposure:

Males Animals have been dosed once a day, 7 days a week, for a minimum of 2 consecutive weeks prior to pairing and thereafter through the day before necropsy. Dose volumes will be adjusted once per week for each animal according to the last recorded body weight.FemalesAnimals will be dosed once a day, 7 days a week, for a minimum of 2 consecutive weeks prior to pairing and thereafter during pairing , post coitum and post partum periods until Day 3 post partum or the day before sacrifice. Dose volumes will be adjusted once per week for each animal according to the last recorded body weight.During the gestation period, dose volumes will be calculated according to individual body weight on Days 0, 7, 14 and 20 post coitum and on Day 1 post partum. Thereafter individual dose volumes will remain constant.

Frequency of treatment:

Once a day

Doses / concentrations

Doses / concentrations 1

Remarks:

Doses / Concentrations:10 mg/Kg bwBasis:actual ingested

Doses / concentrations 2

Remarks:

Doses / Concentrations:100 mg/Kg bwBasis:actual ingested

Doses / concentrations 3

Remarks:

Doses / Concentrations:1000 mg/Kg bwBasis:actual ingested

No. of animals per sex per dose:

10 animals per sex per dose

Control animals:

yes, concurrent vehicle

Details on study design:

The test item have been administered orally by gavage at a dose volume of 10 mL/kg body weight. Control animals received the vehicle alone at the same dose volume.The dose has been administered to each animal on the basis of the most recently recorded body weight and the volume administered has been recorded for each animal

Positive control:

No

Examinations

Observations and examinations performed and frequency:

Mortality Early in each working day in the morning and in the afternoon. At weekends and Public Holidays a similar procedure will be followed except that the final check has been carried out at approximately mid-day. Clinical signsOnce before commencement of treatment and at least once daily during the study, Clinical observations (Functional Observation Battery Tests)Once before commencement of treatment and at least once a week thereafterGrip strength and sensory reactivity to stimuliOnce during the study, towards the end of treatment, on 5 males and 5 females randomly selected from each groupMotor activity assessment (MA)Once during the study, towards the end of treatment, on 5 males and 5 females randomly selected from each groupFood consumptionRecorded weekly (whenever possible) during the pre-mating period starting from allocation. Individual food consumption for the females have been measured on gestation Days 7, 14 and 20 starting from Day 0 post coitum and on Day 4 post partum starting from Day 1 post partum.Body weightMales weekly from allocation to termination.Females weekly from allocation to positive identification of mating and on gestation Days 0, 7, 14 and 20. Dams Days 1 and 4 post partum. Vaginal smears Vaginal smears has been taken daily in the morning starting two weeks before pairing until a positive identification of copulation is made.Mating Parturition and gestation lengthA parturition check has been performed from Day 20 to Day 25 post coitum. FPups identification, weight and observationAs soon as possible, after parturition is considered complete (Day 0 or 1 post partum), .Once daily for all litters

Sacrifice and pathology:

Euthanasia Parental animals in extremis or killed for humane reasons and those that have completed the scheduled test period will be killed by exsanguination under isofluorane anaesthesia.Pups in extremis or killed for humane reasons or those that have completed the scheduled test period (Day 4 post partum) will be euthanised by intraperitoneal injection of Thiopenthal. Parental males: The males will be killed after the mating of all females or after at least 28 days of treatment period.Parental females:The females with live pups will be killed on Day 4 post partum.The females with total litter loss will be killed on the day of the occurrence of total litter loss or shortly after.The females showing no evidence of copulation will be killed after 25 days of the last day of the mating session (see section 4.2.9). The females which do not give birth 25 days after positive identification of mating will be killed shortly after. 4.4.2Necropsy The clinical history of the males and females of the parental generation will be studied and a detailed post mortem examination will be conducted (including examination of the external surface and orifices). Changes will be noted, the requisite organs weighed (excluding animals sacrificed for humane reasons or found dead) and the required tissue samples preserved in fixative and processed for histopathological examination.Females:All females will be examined also for the following:b) number of visible implantation sites (pregnant animals);c) number of corpora lutea (pregnant animals). Uteri of females with no visible implantations will be immersed in a 10-20% solution of ammonium sulphide to reveal evidence of implantation.PupsAll pups found dead in the cage or sacrificed for humane reasons will be examined for external and internal abnormalities.All live pups sacrificed at termination will be killed and examined for external abnormalities and sex confirmation by gonadal inspection.4.4.3Organ weightsFrom all animals completing the scheduled test period, the organs indicated in Annex 1 will be dissected free of fat and weighed.The ratios of organ weight to body weight will be calculated for each animal. At the discretion of the pathologist, organs may be weighed from animals dying or killed prior to terminal kill. 4.4.4Tissues fixed and preserved Samples of all the tissues listed in Annex 1 will be fixed and preserved in 10% neutral buffered formalin (except eyes, optic nerves and Harderian glands, testes and epididymides which will be fixed in modified Davidson's fluid and preserved in 70% ethyl alcohol).4.4.5Histopathological examination The tissues required for histopathological examination are listed in Annex 1. After dehydration and embedding in paraffin wax, sections of the tissues will be cut at 5 micrometer thickness and stained with haematoxylin and eosin. In addition, the testes and epididymides will be cut at 2-3 micrometer thickness and stained with Periodic Acid Schiff (PAS). The morphological evaluation of the seminiferous epithelium (staging of spermatogenic cycle) will be performed. If considered necessary, histological processing may be subcontracted to a GLP certified test site. In such cases, a protocol amendment will be issued, the Sponsor will be informed of the location of the test site and the complete address and name of the Principal Investigator will be presented in the Final Report.In the first instance the examination will be restricted as detailed below: a)Tissues specified in Annex 1 from 5 males and 5 females randomly selected (animals evaluated for clinical pathology) in the control and high dose group killed at term.b)Tissues specified in Annex 1 from all animals killed or dying during the treatment period. c)All abnormalities in all groups. The examination could then be extended to include the remaining 5 males and 5 females (animals not evaluated for clinical pathology) of the control and the high dose group (additional cost). The examination could then be extended to animals of the other dose groups, if treatment-related changes are observed in the high dose group.All histopathological activities which cannot be foreseen before the start of the study (i.e. processing of all abnormalities, tissues of unscheduled deaths and target tissues in the low, medium dose groups) will incur an additional cost.4.4.6PhotomicrographsRepresentative photomicrographs may be taken of any treatment-related lesions. Other photomicrographs may be taken as required by the Sponsor (additional cost).

Other examinations:

As a part of the sacrificial procedure, samples of blood will be withdrawn under isofluorane anaesthesia from the abdominal vena cava from 5 males and 5 females (females with viable litters, if possible) randomly selected from each group, under condition of food deprivation.The blood samples collected will be divided into tubes as follows:EDTA anticoagulantfor haematological investigationsHeparin anticoagulantfor biochemical testsCitrate anticoagulantfor coagulation testsThe measurements to be performed on blood samples are listed below:4.3.1HaematologyHaematocritHaemoglobinRed blood cell countReticulocyte count Mean red blood cell volumeMean corpuscular haemoglobinMean corpuscular haemoglobin concentrationWhite blood cell count Differential leucocyte count- Neutrophils- Lymphocytes- Eosinophils- Basophils- Monocytes- Large unstained cellsPlatelets4.3.2Coagulation testsProthrombin time4.3.3Clinical chemistryAlkaline phosphataseAlanine aminotransferaseAspartate aminotransferaseGamma-glutamyltransferaseUreaCreatinineGlucoseTriglyceridesBile acidsPhosphorusTotal bilirubinTotal cholesterolTotal proteinAlbuminGlobulinA/G RatioSodiumPotassium CalciumChloride4.3.4Urinalysis (Only males – optional, to be discussed see section 3.2) At the same time interval (if required) of the clinical pathology investigations, individual overnight urine samples will also be collected from the same animals under the same conditions. Before starting urine collection water bottles will be removed from each cage and each animal will receive approximately 10 mL/kg of drinking water by gavage, in order to obtain urine samples suitable for analysis.AppearanceVolumeSpecific gravitypHProteinGlucoseTotal reducing substances (only if Glucose is higher than 100 mg/dL)KetonesBilirubinUrobilinogenBloodThe sediment, obtained from centrifugation at approximately 3000 rpm for 10 minutes, will be examined microscopically for:Epithelial cellsLeucocytesErythrocytesCrystalsSpermatozoa and precursorsOther abnormal components

Statistics:

Standard deviations will be calculated as appropriate. For continuous variables the significance of the differences amongst group means will be assessed by Dunnett’s test or a modified t test, depending on the homogeneity of data.Statistical analysis of histopathological findings will be carried out by means of the non-parametric Kolmogorov-Smirnov test if n is more than 5.The non-parametric Kruskal-Wallis analysis of variance will be used for the other parameters. Intergroup differences between the control and treated groups will be assessed by the non-parametric version of the Williams test. The criterion for statistical significance will be p<0.05.Further tests will be used as considered appropriate. Details of all tests used and the data to which they are applied will be included in the Final Report.

Results and discussion

Results of examinations

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:

no effects observed

Water consumption and compound intake (if drinking water study):

no effects observed

Ophthalmological findings:

no effects observed

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

no effects observed

Behaviour (functional findings):

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Effect levels

Dose descriptor:

NOEL

Effect level:

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

Based on:

test mat.

Sex:

male/female

Basis for effect level:

behaviour (functional findings)

body weight and weight gain

clinical biochemistry

clinical signs

food consumption and compound intake

food efficiency

gross pathology

haematology

histopathology: neoplastic

histopathology: non-neoplastic

mortality

ophthalmological examination

organ weights and organ / body weight ratios

urinalysis

water consumption and compound intake

Target system / organ toxicity

Critical effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

NOEL > 1000 mg/kg bw/day