5-sulphosalicylic acid

Endpoint summary

Description of key information

rat, oral, OECD 422, NOAEL = 1000 mg/kg bw/day

Key value for chemical safety assessment

Toxic effect type:

dose-dependent

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

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

Reason / purpose for cross-reference:

reference to other study

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.

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

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.

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

Dose / conc.:

0 mg/kg bw/day (nominal)

Dose / conc.:

100 mg/kg bw/day (nominal)

Dose / conc.:

300 mg/kg bw/day (nominal)

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

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

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

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

Key result

Critical effects observed:

no

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

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

high quality

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

OECD 422

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 groupin concentrations of 10, 30 and 100 mg/mLcorresponding 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

Justification for classification or non-classification

The substance is not classified according to Regulation (EC) No 1272/2008.