5-sulphosalicylic acid

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

18 FEB 2021- 19 OCT 2021 (experimental phase: 09 MAY 2021 - 26 JUN 2021)

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Cross-reference

Reason / purpose for cross-reference:

reference to other study

Reference

Endpoint:

repeated dose toxicity: oral, other

Remarks:

Dose range finding stduy

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

30 APR 2020 - 11 NOV 2020

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Qualifier:

equivalent or similar to guideline

Guideline:

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

Version / remarks:

adopted 29 July 2016.

Deviations:

yes

Principles of method if other than guideline:

Mating period
In the Study Plan: 14 days
Deviation: 16-17 days
Reason for the deviation: The mating period was prolonged for two pairs of animals to ensure the possibility for a successful mating as the female animals were in pseudo-pregnancy state.
Presumed effect on the study: None

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Species:

rat

Strain:

Wistar

Remarks:

WIST of Wistar origin

Details on species / strain selection:

The rat is regarded as suitable species for reproduction studies and the test guideline is designed to use the rat. The Wistar rat was selected due to large experience with this strain of rat in reproduction toxicity studies and known fertility and availability of historical control data at Test Facility.

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Toxi-Coop Zrt. 1103 Budapest, Cserkesz u. 90. Hungary
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: Male animals: 83 – 90 days, Female animals: 83 – 90 days
- Weight at study initiation: 370 – 413 g for male animals, 213 – 242 g for female animals
- Housing: Before mating: 2 animals of the same sex/cage
Mating: 1 male and 1 female / cage
Mated females: individually
Males after mating: 2 animals / cage
Cage type: Type III polypropylene/polycarbonate;
Size: 22 x 32 x 19 cm (width x length x height)
Bedding: Certified laboratory wood bedding (SAFE 3/4-S-FASERN) produced by J. Rettenmaier & Söhne GmbH+Co.KG; D-73494 Rosenberg Holzmühle 1 Germany. The bedding is suitable as nesting material. Details of quality of bedding material were reported. The cages and bedding were changed once or twice a week.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 20 days

DETAILS OF FOOD AND WATER QUALITY:
Animals received ssniff® SSNIFF Rat/Souris Elevage E, 10 mm autoclavable complete feed and ssniff® SM R/M-Z+H complete diet for rats and mice produced by ssniff Spezialdiäten GmbH, D-59494 Soest Germany and tap water, as for human consumption.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 30 - 70 %
- Air changes (per hr): Above 10 air-exchanges/ hour by a central air-condition system.
- Photoperiod (hrs dark / hrs light): Artificial light, from 6 a.m. to 6 p.m.

Route of administration:

oral: gavage

Details on route of administration:

The test item was administered orally via gavage. The route of application was selected in compliance with international guidelines. The oral route is the anticipated route of human exposure to the test item.

Vehicle:

water

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:

VEHICLE
- Concentration in vehicle: 0, 100, 250, 750 mg/kg bw/day
- Amount of vehicle (if gavage): 5 mL/kg bw
- Lot/batch no. (if required): 2003-5525

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Test item content of the dosing formulations was measured on two occasions in the course of the study. Dosing formulations were prepared in Aqua purificata. Samples were diluted with ultrapure water and analysed with a HPLC-UV method.
Test item was quantified in dosing formulations by high performance liquid chromatography coupled with UV detection in the concentration range of 0.05 – 0.25 mg/mL. The dosing formulations were diluted 200×, 500× or 1000× for the analysis.
Linear range: 0.05 – 0.25 mg/mL
inter-day precision of calibrators: ≤ 0.707 %
inter-day accuracy of calibrators: ≤ ± 1.50 %

Duration of treatment / exposure:

All animals of the parent (P) generation were dosed prior to mating (14 days) and throughout mating.
Males: for 34 days. Females: for 35-50 days. Non-pregnant female animals: for 34-38 days.

Frequency of treatment:

Daily
The test item was administered in a single dose by oral gavage on a 7 days/week basis, everyday at a similar time (±2 hours).

Dose / conc.:

0 mg/kg bw/day (nominal)

Dose / conc.:

100 mg/kg bw/day (nominal)

Dose / conc.:

250 mg/kg bw/day (nominal)

Dose / conc.:

750 mg/kg bw/day (nominal)

No. of animals per sex per dose:

6 male, 6 female

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: The dose levels were chosen in agreement with the Sponsor taking into consideration the acute oral LD50 value of 5-Sulfosalicylic acid from literary data (rat oral LD50: 1850 mg/kg bw; National Library of Medicine; PubChem).
The high dose was chosen with the aim of inducing toxic effects but no mortality or severe suffering of animals or representing the limit dose according to the respective OECD guideline. The low dose was chosen to induce no toxic effect. The mid dose is interpolated geometrically.
- Rationale for animal assignment (if not random): Animals were selected for this study on the basis of adequate body weight, a body weight within ± 20% of the mean within a sex and free from clinical signs of disease or injury. Selected rats were distributed by randomization according to stratification by body weight so that there was no statistically significant difference among group body weight means within a sex.

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: once a day

DETAILED CLINICAL OBSERVATIONS: Yes

BODY WEIGHT: Yes
- Time schedule for examinations: Parental females were weighed on the first day of dosing (Day 0) then weekly, on gestation days 0, 7, 14 and 19. Body weight of the female animals was additionally weighed on gestational day 10 in order to give accurate treatment volumes

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
The food consumption was determined weekly by reweighing the given and non-consumed diet with a precision of 1 g during the treatment period except mating phase (pre-mating days 0, 7, 13, and post-mating days 20, 27 and 33 for male animals; pre-mating days 0, 7, 13, gestation days 0, 7, 14 and 19 for female animals).

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Not specified

HAEMATOLOGY: Yes
- Time schedule for collection of blood: one day after the last treatment
- Anaesthetic used for blood collection: Yes (identity)
- Animals fasted: Yes, Animals were food deprived for approximately 16 hours (overnight) prior to blood collection.:
- Parameters examined.: WBC, RBC, HGB, HCT, MCV, MCH, MCHC, PLT, RET, Different withe blood cell count

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: one day after the last treatment
- Animals fasted: Yes, Animals were food deprived for approximately 16 hours (overnight) prior to blood collection.
- Parameters examined.: ALT, AST, TBIL, CREA, UREA, GLUC, CHOL, Na+

PLASMA/SERUM HORMONES/LIPIDS: Not specified

URINALYSIS: Not specified

NEUROBEHAVIOURAL EXAMINATION: Not specified

IMMUNOLOGY: Not specified

Sacrifice and pathology:

GROSS PATHOLOGY: Yes

HISTOPATHOLOGY: Yes

Clinical signs:

no effects observed

Description (incidence and severity):

The behavior and physical condition of animals were considered to be normal in all male and female animals in the control, 100, 250 and 750 mg/kg bw/day groups during the daily clinical observations.

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

Statistical significance with respect to the control was detected at the slightly higher mean daily food consumption of male animals at 750 mg/kg bw/day between Days 27 and 33. This minor difference with respect to the control has no toxicological relevance.

Food efficiency:

not specified

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

not specified

Ophthalmological findings:

not specified

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Description (incidence and severity):

Statistical significance noted for the slightly lower mean potassium concentration (K+) in the male animals at 250 and 750 mg/kg bw/day was considered to be not related to the treatment or test item. The range of values at 250 and 750 mg/kg bw/day were comparable with the actual control although were slightly below the historical control.

Endocrine findings:

not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

no effects observed

Description (incidence and severity):

The behavior and physical condition of animals were considered to be normal in all male and female animals in the control, 100, 250 and 750 mg/kg bw/day groups during the daily clinical observations.

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

Statistically significant differences with respect to the control in male animals– lower mean liver weights (absolute and relative to body and brain weights) at 250 mg/kg bw/day and higher mean kidney weight relative to the body weight at 100 mg/kg bw/day– were incidental and not treatment related changes. Similarly, in the female animals a 250 mg/kg bw/day, a slightly higher mean brain weight relative to corrected body weight and a lower mean corrected body weight relative to brain weight were judged to be not related to the treatment as similar finding was not detected at the higher dose group.

Gross pathological findings:

no effects observed

Description (incidence and severity):

There were no signs of inflammation or other pathological changes, therefore pyelectasia and hydrometra were considered to be toxicologically not relevant. Hernia diaphragmatica is also frequently observed macroscopic findings in experimental rats without toxicological significance. Hemorrhages in the stomach was considered to be an individual alteration in single animal at 250 mg/kg bw/day regarding the low incidence and because similar finding was not observed in the higher dose administered animals.

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

Dilatation of uterine horns (in connection with the “hydrometra”) in one non-pregnant female animal (1/2 at 100 mg/kg bw/day) without degenerative or inflammatory lesions, is considered as a slight neuro-hormonal phenomenon in connection with estrus cycle.

Histopathological findings: neoplastic:

no effects observed

Description (incidence and severity):

In the organs of animals subjected to histopathological examination, no lesions appearing only in the treated animals in function of applied doses, thus unanimously attributable to the test item were detected.

Other effects:

no effects observed

Description (incidence and severity):

Estrous cycle: Test item influence on the estrous cycle was not detected at any dose level (100, 250 and 750 mg/kg bw/day).
Pregnancy data of dams: There were no test item related differences in the evaluated pregnancy parameters of dams between the control and test item treated groups (100, 250 or 750 mg/kg bw/day).
Reproductive performance: The examined parameters of reproductive performance were not affected by the treatment with the test item in male or female animals at 100, 250 or 750 mg/kg bw/day.

Dose descriptor:

NOAEL

Effect level:

>= 750 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no adverse effects in highest dose group

Critical effects observed:

no

Conclusions:

Based on these observations the No Observed Adverse Effect Levels (NOAEL) were determined as follows:
NOAEL for systemic toxicity of male/female rats: 750 mg/kg bw/day
NOAEL for reproductive performance of male/female rats: 750 mg/kg bw/day
NOAEL for F1 offspring development: 750 mg/kg bw/day
The suggested doses for an OECD 422 study are as follows:
Control (0 mg/kg bw/day)
100 mg/kg bw/day
300 mg/kg bw/day
1000 mg/kg bw/day

Executive summary:

Under the conditions of the present study, test item did not cause signs of systemic toxicity in parental male and female Han:Wistar rats at 100, 250 or 750 mg/kg bw/day doses administered by oral gavage and the test item did not adversely influence the reproductive performance (gonad function, mating behavior, conception) in parental male or female rats at the doses of 100, 250 or 750 mg/kg bw/day.

The development of the fetuses was not impaired at any dose level from conception to gestation day 19 after repeated oral administration of dams.

Based on these observations the No Observed Adverse Effect Levels (NOAEL) were determined as follows:

NOAEL for systemic toxicity of male/female rats: 750 mg/kg bw/day

NOAEL for reproductive performance of male/female rats: 750 mg/kg bw/day

NOAEL for F1 offspring development: 750 mg/kg bw/day

The suggested doses for an OECD 422 study are as follows:

Control (0 mg/kg bw/day)

100 mg/kg bw/day

300 mg/kg bw/day

1000 mg/kg bw/day

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2021

Report date:

2021

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

adopted 29 July 2016.

Deviations:

yes

Remarks:

Mating period was prolonged by a day to ensure a successful mating for two pairs at 100 mg/kg bw/day.

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: EPA Health Effects Test Guidelines: OPPTS 870.3650 Combined Repeated Dose Toxicity with the Reproduction/Developmental Toxicity Screening Test

Version / remarks:

July 2000

Deviations:

yes

Remarks:

Mating period was prolonged by a day to ensure a successful mating for two pairs at 100 mg/kg bw/day.

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

5-sulphosalicylic acid

EC Number:

202-555-6

EC Name:

5-sulphosalicylic acid

Cas Number:

97-05-2

Molecular formula:

C7H6O6S

IUPAC Name:

2-hydroxy-5-sulfobenzoic acid

Test material form:

solid: crystalline

Details on test material:

Batch number 1: 1907015
Batch number 2: 2005014
CAS number: 5965-83-3
EC number 202-555-6
Molecular formula: C7H6O6Sx2H2O
Appearance: White to off white crystals
Assay: 101.9 % – batch 1
102.2 % – batch 2
Manufacturing date. July 15, 2019 – batch 1
May 01, 2021 – batch 2
Expiry date: July 31, 2021 – batch 1
April 30, 2022 – batch 2
Storage conditions: At room temperature (15 – 25°C); protected from light
Safety precautions: According to safety data sheet

Test animals

Species:

rat

Strain:

Wistar

Remarks:

Han:WIST of Wistar origin

Details on species / strain selection:

The rat is regarded as suitable species for reproduction studies and the test guideline is designed to use the rat. The Wistar rat was selected due to large experience with this strain of rat in reproduction toxicity studies and known fertility.

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Toxi-Coop Zrt. 1103 Budapest, Cserkesz u. 90. Hungary
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 82 –89 days
- Weight at study initiation: 348 – 402 g for male animals, 211 – 261 g for female animals
- Housing: Before mating: 2 animals of the same sex/cage
Mating: 1 male and 1 female / cage
Mated females: individually
Males after mating: 2 animals / cage
- Diet (e.g. ad libitum): ad libitum. ssniff® SM R/M-Z+H complete diet for rats and mice. Food was changed at weekly intervals.
- Water (e.g. ad libitum): ad libitum. changed daily
- Acclimation period: 19 days

DETAILS OF FOOD AND WATER QUALITY:
The food was considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study. The supplier provided an analytical certificate of the standard diet for the batch used.
Water quality control analysis and microbiological assessment are performed once in every six months by Government Office of Capital Budapest Department of Public Health and Medical Officer Service (Váci út 172-174. Budapest, H-1138 Hungary).

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 30 - 70 %
- Air changes (per hr): Above 10 air-exchanges/ hour by a central air-condition system.
- Photoperiod (hrs dark / hrs light): Artificial light, from 6 a.m. to 6 p.m.

Administration / exposure

Route of administration:

oral: gavage

Details on route of administration:

The test item was administered orally via gavage. The route of application was selected in compliance with international guidelines. The oral route is the anticipated route of human exposure to the test item.

Vehicle:

water

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:

VEHICLE
- Concentration in vehicle: 10, 30 and 100 mg/mL
- Amount of vehicle (if gavage): 10 mL/kg bw

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Five aliquots of 5 mL of each formulation and five aliquots of control substance (vehicle) were taken two times and were analyzed.
Date of sampling: March 17 and April 22, 2021
Date of analysis: March 17 and April 23, 2021
Concentration of the test item in the dosing formulations varied between the range of 98.3 – 105 % in comparison to the nominal values.

5-Sulfosalicylic acid was quantified in dosing formulations by high performance liquid chromatography coupled with UV detection in the concentration range of 0.05 – 0.25 mg/mL.
The dosing formulations were diluted 100×, 200× or 1000× for the analysis.
Linear range: 0.05-0.25 mg/mL
Limit of Quantification 0.05 mg/mL
Recovery: 93.2 % (1 mg/mL); 95.3 % (150 mg/mL)
Selectivity: No interfering component was detected in the blank sample (ultrapure water) and in the blank formulation (Aqua purificata)

Duration of treatment / exposure:

Male: From Day 0 up to Day 49, (March 09 – April 27, 2021), daily up to the day before the necropsy (during the pre-mating, mating and post-mating periods).
Female: From Day 0 up to Day 63, (March 09 – May 11, 2021), daily up to the day before the necropsy (during the pre-mating, mating, gestation and lactation periods).

Frequency of treatment:

administered orally (by gavage) once daily

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

0 mg/kg bw/day (nominal)

Doses / concentrations 2

Dose / conc.:

100 mg/kg bw/day (nominal)

Doses / concentrations 3

Dose / conc.:

300 mg/kg bw/day (nominal)

Doses / concentrations 4

Dose / conc.:

1 000 mg/kg bw/day (nominal)

No. of animals per sex per dose:

12 animals per sex per dose
12 animals per sex for control group

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: The dose levels were chosen on the basis of the results of a preliminary dose range finding study with 5-Sulfosalicylic Acid in rats (Study no. 968-400-5401).
- Rationale for animal assignment (if not random): Selected rats were distributed by randomization according to stratification by body weight so that there was no statistically significant difference among group body weight means within a sex.
- Fasting period before blood sampling for clinical biochemistry: Animals were food deprived for approximately 16 hours (overnight) prior to blood collection.

Positive control:

no

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Day 0, (March 09, 2021), then weekly and on the day before necropsy;

BODY WEIGHT: Yes
- Time schedule for examinations: Male: From Day 0 up to Day 48, (March 09 – April 26, 2021), weekly (during the pre-mating, mating and post-mating periods)
Female: From Day 0 (March 09), weekly prior to and during the mating period. On gestational days 0, 7, 10, 14 and 21. On post-partum days 0, 4 and 13
Body weight of parental animals selected for organ weighing were determined on the day of necropsy

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
Male: From Day 0 up to Day 48, (March 09 – April 26, 2021), weekly prior to and after the mating period.
Female: From Day 0 (March 09, 2021), weekly prior to and after the mating period. Also on gestational days 0, 7, 14, and 21 and on post-partum days 0, 4 and 13

The mean daily food consumption was comparable in the control and 100, 300 and 1000 mg/kg bw/day during the entire observation period (pre-mating and post-mating periods for male animals, pre-mating, gestation and lactation period for female animals).

OPHTHALMOSCOPIC EXAMINATION: Not specified

HAEMATOLOGY: Yes
- Time schedule for collection of blood:
Sires (male animals): Day 50, April 28, 2021 (day of necropsy)
Dams: Day 52, April 30, 2021 (day of necropsy)
- Anaesthetic used for blood collection: Yes (Isofluran CP)
- Animals fasted: Yes
- How many animals: 5 males and 5 females

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: on the day of necropsy
- Animals fasted: Yes
- How many animals: 5 males and 5 females

PLASMA/SERUM HORMONES/LIPIDS: Yes
- Time of blood sample collection: from 2-7 pups per litter on post-natal day 4 (in litters with at least 10 pups; samples were pooled by litter)
from all dams and from 3-7 pups per litter on post-partum/post-natal day 13;
from all parent male animals at termination on Day 49.
- Animals fasted: Yes
- How many animals: all parent animals, 2-7 pups per litter

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No

IMMUNOLOGY: No

Sacrifice and pathology:

GROSS PATHOLOGY: Yes

HISTOPATHOLOGY: Yes

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Description (incidence and severity):

Alopecia, scars and porphyrin around the eye – as species specific findings of this species and strain of rat with similar age – was noted with low incidence.

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Description (incidence and severity):

Statistical significance was only detected at the slightly higher mean body weight gain in male animals at 100 mg/kg bw/day between Days 27 and 34.
This minor change was were considered to be toxicologically not relevant.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

Statistical significance was noted for the slightly higher mean daily food consumption in male animals at 300 mg/kg bw/day comparing to their control between Days 27 and 34 and between Days 34 and 41.
This minor difference was considered to be indicative of biological variation and not related to test item administration.

Food efficiency:

not specified

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

not specified

Ophthalmological findings:

not specified

Haematological findings:

no effects observed

Description (incidence and severity):

Statistically significant difference with respect their control was detected at the higher mean platelet count (PLT) in male animals at 100 and 300 mg/kg bw/day independently from doses. In the female animals at 100 and 300 mg/kg bw/day, statistical significance with respect to the control was observed at the slightly lower mean percentage of monocytes (MONO), at the lower mean corpuscular hemoglobin content (MCH) and at the lower mean corpuscular volume (MCV), when compared to the control. At 1000 mg/kg bw/day, the mean corpuscular hemoglobin content, mean corpuscular volume and prothrombin time (PT) were lower than in the control group in the female animals. All these changes were considered to be variations and toxicologically not relevant due to the minor degree or the lack of dose dependency (PLT, MONO, MVH, MCHC and PT). Individual values were within the historical control ranges except for PLT of one male animal at 100 mg/kg bw/day. Therefore, these findings were judged to be accidental.

Clinical biochemistry findings:

no effects observed

Description (incidence and severity):

Statistical significance noted for the slightly lower mean glucose (GLUC) concentration in the male animals at 1000 mg/kg bw/day was considered to be not related to the treatment or test item. The individual values of glucose level were well within the historical control range. In the female animal, all examined clinical chemistry parameters were comparable with the control at 100, 300 and 1000 mg/kg bw/day.

Endocrine findings:

not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

no effects observed

Description (incidence and severity):

There were no changes in the physical condition, behavior or in reactions to different types of stimuli in the selected male or female animals of control and test item treated groups in the examined parameters during the course of the functional observations.

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

Some minor but statistically significant difference with respect to the control was detected in male animals at 1000 mg/kg bw/day at the lowered mean heart weight (absolute and relative to body weight) and at the lowered mean weight of seminal vesicle with coagulating gland and prostate as a whole (absolute). Both changes were minimal, the changes for absolute and relative weights were well within the historical control range and no histopathological alterations were detected. The weights of all examined organs (absolute and relative to body or brain weights) were similar in the female animals in control and 100, 300 and 1000 mg/kg bw/day groups.

Gross pathological findings:

no effects observed

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

Histopathological investigations did not reveal test item related lesions in the organs or tissues of animals selected for toxicity examinations at 1000 mg/kg bw/day (male and female).

Histopathological findings: neoplastic:

no effects observed

Other effects:

not specified

Effect levels

Key result

Dose descriptor:

NOAEL

Effect level:

>= 1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no adverse effects up to highest dose

Target system / organ toxicity

Key result

Critical effects observed:

no

Applicant's summary and conclusion

Conclusions:

Under the conditions of the present study, 5-Sulfosalicylic Acid administered at 100, 300 and 1000 mg/kg bw/day oral gavage did not show adverse systemic toxicity in parental male and female Han:WIST rats.Based on these observations the No Observed Adverse Effect Levels (NOAEL) were determined as follows:
NOAEL for systemic toxicity of male/ female parental rats: 1000 mg/kg bw/day
NOAEL for reproductive performance of male/ female parental rats: 1000 mg/kg bw/day

Executive summary:

A repeat dose toxicity test was performed according to OECD Guideline 422 and in compliance with GLP. 5-Sulfosalicylic Acid was administered orally (by gavage) once daily at 0 (vehicle only), 100, 300 and 1000 mg/kg bw/d doses to four groups of Han:WIST rats consisting of 12 animals per sex per group in concentrations of 10, 30 and 100 mg/mL corresponding to a 10 mL/kg bw dosing volume. A group of vehicle (distilled water) treated animals (n= 12/sex) served as a control. Under the conditions of the present study, 5-Sulfosalicylic Acid administered at 100, 300 and 1000 mg/kg bw/day oral gavage did not show adverse systemic toxicity in parental male and female Han:WIST rats.

Based on these observations the No Observed Adverse Effect Levels (NOAEL) were determined as follows:

NOAEL for systemic toxicity of male/ female parental rats: 1000 mg/kg bw/day

NOAEL for reproductive performance of male/ female parental rats: 1000 mg/kg bw/day