Tosylchloramide sodium

Endpoint summary

Description of key information

There are several studies available for evaluation of which the 90-day in dogs, rats and mice are the longest in duration.

Most relevant:

Toxicty to reproduction study oral in rats 2 -generation study in rats (OECDTG 416): NOAEL 65 mg/kgbw/d (based on 1000 ppm in diet) due to transient effects on body weights at 3000 ppm (read-across from p-TSA).

Other available data:

Repeated dose toxicity oral (OECDTG 408) with p-TSA: NOAEL 90-d rat = 3000 ppm (214 mg/kg/day for males; 248 mg/kg/day for females).

Repeated dose toxicity oral (OECDTG 409) with p-TSA: NOAEL 90-d dog = 8000 ppm (260 mg/kg/day for males; 255 mg/kg/day for males).

Toxicity to reproduction screening study (OECD 422, oral-rat with p-TSA): NOAEL of < 120 mg/kg bw/d.

Repeated dose toxicity dermal in rabbits with Halamid (21 -day, comparable to guideline study with acceptable restrictions) :

NOEL for systemic effects is 225 mg/kg bw/day

NOEL for effects on the skin is 25 mg/kg bw/day.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Reference 1

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2006 October 25 - 2007 January 30

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Study performed according to EC and OECD guidelines and according to GLP principles.

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)

Deviations:

yes

Remarks:

See below.

Qualifier:

according to guideline

Guideline:

EU Method B.26 (Sub-Chronic Oral Toxicity Test: Repeated Dose 90-Day Oral Toxicity Study in Rodents)

Deviations:

yes

Remarks:

See below.

Principles of method if other than guideline:

Deviations:
1. One adrenal from animal no. 67 and from animal no. 4 was not available for histopathology. Reasons for this included that these tissues were not discernable at necropsy or trimming, or were erroneously not collected at necropsy. Tissues are listed in raw data and pathology report. Evaluation: Sufficient tissues were available for evaluation.
2. No terminal body weight was determined from animal no. 55, and no adrenal weight was recorded for animal no. 4. Evaluation: Sufficient data were available for evaluation.

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Species:

rat

Strain:

other: Wistar Han, Crl:WI(Han)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: Approximately 6 weeks
- Housing: 5 animals/sex in Macrolon cages with sterilised saw dust as bedding material and paper as cage-enrichment.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 5 days.


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.0-23.2
- Humidity (%): 35-87
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

DIET PREPARATION
- Rate of preparation of diet (frequency): diets were prepared for weeks 1-4, 5-8 and 9-13.
- Mixing appropriate amounts with (Type of food): Standard powder rodent diet

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The concentrations analysed in the diets were between 83 and 113% of target. In the control diet, no test substance was detected.

Duration of treatment / exposure:

At least 90 days.

Frequency of treatment:

Ad libitum

Remarks:

Doses / Concentrations:
1000 ppm
Basis:
nominal in diet

Remarks:

Doses / Concentrations:
3000 ppm
Basis:
nominal in diet

Remarks:

Doses / Concentrations:
10000 ppm
Basis:
nominal in diet

No. of animals per sex per dose:

10

Control animals:

yes, plain diet

Details on study design:

- Dose selection rationale: Based on results of a 28-day range finding study

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes, mortality
- Time schedule: at least twice daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: at least once daily

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No
- Time schedule for examinations:

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: pre-test and week 13
- Dose groups that were examined: all animals, animals from groups 1 and 4, respectively

HAEMATOLOGY: Yes
- Time schedule for collection of blood: week 13
- Anaesthetic used for blood collection: Yes, iso-flurane
- Animals fasted: Yes
- How many animals: all animals
- Parameters examined: Erythrocytes count, haemoglobin, Haematocrit, Mean corpuscular volume, Mean corpuscular haemoglobin, Mean corpuscular haemoglobin concentration, Platelet count, Red cell distribution width, Total leucocytes count, Differential leucocyte count, Prothrombin time, Partial thromboplastin time.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: week 13
- Animals fasted: Yes
- How many animals: all animals
- Parameters examined: Alanine aminotransferase, Alkaline phosphatase, Aspartate aminotransferase, Bilirubin, total, Chloride, Cholesterol, total, Creatinine, Glucose, Phosphorus, Protein, total, Protein, albumin, Urea, Calcium, Potassium, Sodium.

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: week 12-13
- Dose groups that were examined: all
- Battery of functions tested: grip strength / motor activity / other: hearing ability, pupillary reflex, static righting reflex.

Sacrifice and pathology:

Organ weights: Adrenal glands, Ovaries, Brain, Spleen, Epididymides, Testes, Heart, Thymus, Kidneys, Uterus, Liver.
GROSS PATHOLOGY: Yes (see table)
HISTOPATHOLOGY: Yes (see table)

Other examinations:

None.

Statistics:

None.

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, treatment-related

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

no effects observed

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

not examined

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:

effects observed, treatment-related

Histopathological findings: neoplastic:

no effects observed

Details on results:

BODY WEIGHT AND WEIGHT GAIN
Body weights and body weight gain were reduced in males and females at 10.000 ppm, achieving a level of statistical significance on most occasions. Total weight gain deficit over the treatment period compared to controls was 21% and 11% for males and females, respectively.
Body weights and body weight gain of animals at 1000 or 3000 ppm remained in the same range as controls over the study period.

HISTOPATHOLOGY: NON-NEOPLASTIC
In 2/10 males at 10.000 ppm, a minimal degree of hyperplasia of the urothelium of the urinary bladder was recorded.

Key result

Dose descriptor:

NOAEL

Effect level:

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

Based on:

test mat.

Sex:

male

Basis for effect level:

other: Reduced body weight gain.

Key result

Dose descriptor:

NOAEL

Effect level:

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

Based on:

test mat.

Sex:

female

Basis for effect level:

other: Reduced body weight gain.

Key result

Dose descriptor:

LOAEL

Effect level:

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

Based on:

test mat.

Sex:

male

Basis for effect level:

other: Reduced body weight gain, histopathology (minimal urethelial hyperplasia in two animals)

Key result

Dose descriptor:

LOAEL

Effect level:

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

Based on:

test mat.

Sex:

female

Basis for effect level:

other: Reduced body weight gain.

Dose descriptor:

NOAEL

Effect level:

3 000 ppm

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Reduced body weight gain, histopathology (minimal urethelial hyperplasia in two animals)

Dose descriptor:

LOAEL

Effect level:

10 000 ppm

Based on:

test mat.

Sex:

male/female

Basis for effect level:

body weight and weight gain

other: Reduced body weight gain.

Critical effects observed:

not specified

Conclusions:

LOAEL: 10.000 ppm (738 mg/kg/day for males, or 795 mg/kg/day for females).
NOAEL: 3000 ppm (214 mg/kg/day for males, or 248 mg/kg/day for females).

Executive summary:

The study was based on the following guidelines.

- EC Directive 67/548/EEC, B Repeated Dose (90 days) Toxicity (oral), 2001.

- OECD 408, Repeated Dose 90-day Oral Toxicity Study in Rodents, 1998.

- EPA 712-C-98-199, 90-Day Oral Toxicity, 1998.

Based on a 28-day dietary range finding study (NOTOX Project 474053) and in consultation with the sponsor, the dose levels for this 90-day dietary study were selected to be 0, 1000, 3000 and 10.000 ppm. SPF-bred Wistar Han rats received the test substance by dietary intake for at least 90 days. One control group and three treated groups were tested, each consisting of 10 males and 10 females.

Chemical analyses of diet preparations were conducted to assess accuracy and homogeneity of diet preparations.

The following parameters were evaluated: Clinical signs (daily), functional observations (week 13), body weight and food consumption (weekly), and ophthalmoscopy (pretest and week 13). At termination: clinical pathology, macroscopy, organ weights and histopathology on a selection of tissues.

RESULTS

Homogeneity and accuracy of diet preparations were demonstrated by analyses.

Dietary inclusion levels of 0, 1000, 3000 and 10.000 ppm were equivalent to an average test article intake of 0, 70, 214 and 738 mg/kg body weight/day for males, and 0, 80, 248 and 795 mg/kg body weight/day for females.

At 10.000 ppm, a significant and consistent reduction of body weight gain was recorded (up to 21% compared to control animals).

Histopathology revealed a minimal degree of hyperplasia of the urothelium of the urinary bladder in two males at 10.000 ppm.

No other treatment-related toxicologically significant changes were noted in any of the remaining parameters examined/determined in this study (i.e. clinical appearance, functional observations, food consumption, clinical laboratory investigations, macroscopic examination and organ weights).

The observed minimal urethelial hyperplasia in two animals seems to be caused by local irritation by the chronic availability of p-TSA metabolite (p-sulphamonylbenzoic acid) in the urine, following the rapid and complete excretion of p-TSA. Such effect is local and fully reversible when exposure ends. As the hyperplasia is observed in only two animals, is minimal and likely a fully reversible temporal effect upon exposure, is not an adverse effect persé, the reduction of the body weight gain of 21% in males is considered serious enough to assign the 10000 ppm a LOAEL. Especially as a relation to palatability is not completely clear.

LOAEL: 10.000 ppm (738 mg/kg/day for males, or 795 mg/kg/day for females).

NOAEL: 3000 ppm (214 mg/kg/day for males, or 248 mg/kg/day for females).

These results were used for read-across to Tosylchloramide Sodium.

Reference 2

Endpoint:

sub-chronic toxicity: oral

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Justification for type of information:

The full read across justification report is attached under "Attached justification".

Summary

Within REACH, the obligation to conduct tests with vertebrate animals should be considered as a last resort, only after exhausting all potential sources of information on the physical and (eco)toxicological properties of chemicals [1]. Article 13 of REACH requires that use must be made whenever possible by alternatives to vertebrate animal tests, through the use of alternative methods as in vitro methods, (Q)SARs or from structurally related substances via grouping or read-across.
Annex XI of REACH offers the option to evaluate specific endpoints by read-across. According to Chapter R.6 (QSARs and grouping of chemicals) of REACH technical guidance documents (TGD), read-across can be applied for “substances whose physico-chemical, toxicological and ecotoxicological properties are likely to be similar or follow a regular pattern as a result of structural similarity”. Application of the read-across concept requires that physico-chemical properties, human health effects and environmental effects or environmental fate may be predicted from data of a reference substance to make a prediction for the endpoint of a target chemical. This avoids the need to test every substance for every endpoint. This report provides the justification to use a cross-reading approach between Toluenesulfonamides (p-Toluenesulfonamide (p-TSA, CAS 70-55-3), o-Toluenesulfonamide (o-TSA, CAS 88-19-7) and o/p-Toluenesulfonamide (o/p-TSA, CAS 1333 07 9)) and is presented according to the requirements stipulated in Chapter R.6 (QSARs and grouping of chemicals) of the REACH TGD for N-chloro-4-methylbenzenesulfonamide sodium (Chloramine-T, CAS 127-65-1).

Reason / purpose for cross-reference:

read-across source

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, treatment-related

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

no effects observed

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

not examined

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:

effects observed, treatment-related

Histopathological findings: neoplastic:

no effects observed

Details on results:

BODY WEIGHT AND WEIGHT GAIN
Body weights and body weight gain were reduced in males and females at 10.000 ppm, achieving a level of statistical significance on most occasions. Total weight gain deficit over the treatment period compared to controls was 21% and 11% for males and females, respectively.
Body weights and body weight gain of animals at 1000 or 3000 ppm remained in the same range as controls over the study period.

HISTOPATHOLOGY: NON-NEOPLASTIC
In 2/10 males at 10.000 ppm, a minimal degree of hyperplasia of the urothelium of the urinary bladder was recorded.

Key result

Dose descriptor:

NOAEL

Effect level:

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

Based on:

test mat.

Sex:

male

Basis for effect level:

other: Reduced body weight gain.

Key result

Dose descriptor:

NOAEL

Effect level:

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

Based on:

test mat.

Sex:

female

Basis for effect level:

other: Reduced body weight gain.

Key result

Dose descriptor:

LOAEL

Effect level:

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

Based on:

test mat.

Sex:

male

Basis for effect level:

other: Reduced body weight gain, histopathology (minimal urethelial hyperplasia in two animals)

Key result

Dose descriptor:

LOAEL

Effect level:

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

Based on:

test mat.

Sex:

female

Basis for effect level:

other: Reduced body weight gain.

Dose descriptor:

NOAEL

Effect level:

3 000 ppm

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Reduced body weight gain, histopathology (minimal urethelial hyperplasia in two animals)

Dose descriptor:

LOAEL

Effect level:

10 000 ppm

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Reduced body weight gain.

Critical effects observed:

not specified

Conclusions:

LOAEL: 10.000 ppm (738 mg/kg/day for males, or 795 mg/kg/day for females).
NOAEL: 3000 ppm (214 mg/kg/day for males, or 248 mg/kg/day for females).

Executive summary:

The study was based on the following guidelines.

- EC Directive 67/548/EEC, B Repeated Dose (90 days) Toxicity (oral), 2001.

- OECD 408, Repeated Dose 90-day Oral Toxicity Study in Rodents, 1998.

- EPA 712-C-98-199, 90-Day Oral Toxicity, 1998.

Based on a 28-day dietary range finding study (NOTOX Project 474053) and in consultation with the sponsor, the dose levels for this 90-day dietary study were selected to be 0, 1000, 3000 and 10.000 ppm. SPF-bred Wistar Han rats received the test substance by dietary intake for at least 90 days. One control group and three treated groups were tested, each consisting of 10 males and 10 females.

Chemical analyses of diet preparations were conducted to assess accuracy and homogeneity of diet preparations.

The following parameters were evaluated: Clinical signs (daily), functional observations (week 13), body weight and food consumption (weekly), and ophthalmoscopy (pretest and week 13). At termination: clinical pathology, macroscopy, organ weights and histopathology on a selection of tissues.

RESULTS

Homogeneity and accuracy of diet preparations were demonstrated by analyses.

Dietary inclusion levels of 0, 1000, 3000 and 10.000 ppm were equivalent to an average test article intake of 0, 70, 214 and 738 mg/kg body weight/day for males, and 0, 80, 248 and 795 mg/kg body weight/day for females.

At 10.000 ppm, a significant and consistent reduction of body weight gain was recorded (up to 21% compared to control animals).

Histopathology revealed a minimal degree of hyperplasia of the urothelium of the urinary bladder in two males at 10.000 ppm.

No other treatment-related toxicologically significant changes were noted in any of the remaining parameters examined/determined in this study (i.e. clinical appearance, functional observations, food consumption, clinical laboratory investigations, macroscopic examination and organ weights).

The observed minimal urethelial hyperplasia in two animals seems to be caused by local irritation by the chronic availability of p-TSA metabolite (p-sulphamonylbenzoic acid) in the urine, following the rapid and complete excretion of p-TSA. Such effect is local and fully reversible when exposure ends. As the hyperplasia is observed in only two animals, is minimal and likely a fully reversible temporal effect upon exposure, is not an adverse effect persé, the reduction of the body weight gain of 21% in males is considered serious enough to assign the 10000 ppm a LOAEL. Especially as a relation to palatability is not completely clear.

LOAEL: 10.000 ppm (738 mg/kg/day for males, or 795 mg/kg/day for females).

NOAEL: 3000 ppm (214 mg/kg/day for males, or 248 mg/kg/day for females).

Reference 3

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

2007 August 3 - 2007 November 30

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Information derived from an analogue

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 409 (Repeated Dose 90-Day Oral Toxicity Study in Non-Rodents)

Deviations:

yes

Qualifier:

according to guideline

Guideline:

EU Method B.27 (Sub-Chronic Oral Toxicity Test: Repeated Dose 90-Day Oral Toxicity Study in Non-Rodents)

Deviations:

yes

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.3150 (90-Day Oral Toxicity in Non-rodents)

Deviations:

yes

Principles of method if other than guideline:

Protocol deviations:
1 Temporary deviations from the maximum level of relative humidity occurred. Evaluation: Laboratory historical data do not indicate an effect of the deviations.
2 Diets prepared for use in weeks 1-2 and 3-4 were dried for approximately 2 and 3 days respectively, instead of approximately 24 hours. Evaluation: Based on food intake and analytical results of diets prepared for use in week 3-4, this was considered to have no adverse effect on the quality of the diets.
3 Inadvertently, no food consumption was determined for animal no. 7 between days 16-17. Evaluation: deviation was of an incidental nature. Sufficient food consumption data were available for evaluation.
4 On the following days, the following dogs were inadvertently separated on the day following their group housing: day 16 (nos. 30 and 31), day 43 (nos. 29-30), day 60 (nos. 27-28) and day 70 (nos. 23 and 24). Evaluation: Food intake data on these days were excluded, since these were considered not to be representative. Sufficient food intake data were available for evaluation.
5 V3286 (treatment) or V3278 (pretest) was used as batch for SSNIFF® Hd Ereich, Extrudat. Evalua tion: Both batches were valid at the time of use.
6 Lacrimal glands from animal no. 32 were not available for histopathology. Evaluation: Sufficient data was available for evaluation. The study integrity was not adversely affected by the deviations.

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Species:

dog

Strain:

Beagle

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Marshall BioResources, Green Hill 2001, Via San Zeno, 25018 Montichiari (BS), Italy.
- Age at study initiation: Approximately 5-6 months
- Weight at study initiation: males 7.1 - 9.4 kg; females 6.1 - 8.2 kg
- Fasting period before study: not applicable
- Housing: Animals were housed in stainless steel cages (dimensions 143 x 88 x 106 cm) with a
resting shelf (dimensions 61 x 46 cm) and Tenderfoot floors. The animals were group-housed per dos
e group/sex for at least 3 hours per day, on weekdays only.
- Diet (e.g. ad libitum): Animals were offered the test diet once daily at 0.300 kg/animal/day.
- Water (e.g. ad libitum): Ad libitum
- Acclimation period: Approximately 3 weeks.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18.0 - 22.6°C
- Humidity (%): 33 - 97%. Temporary deviations from the maximum level of relative humidity occurred. Laboratory historical data do not indicate an effect of the deviations.
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS: The test substance was dissolved in acetone (± 18-20 ml acetone per kg diet) and then mixed with some powder feed (premix). Subsequently, this premix was mixed with the bulk of the diet. Elix water (approximately 25% in total) was added to aid pelleting. The pellets were dried for approximately 24 hours at 35°C before storage.

DIET PREPARATION
- Rate of preparation of diet (frequency): Diets were prepared every two weeks, or sooner.
- Mixing appropriate amounts with (Type of food): Powder diet for dogs (SSNIFF® Hd Ereich, V3286 (treatment) or V3278 (pretest), Extrudat; SSNIFF® Spezialdiäten GmbH, Soest, Germany).

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Analysis of the accuracy diet preparations revealed values within the range of 80-99% of nominal,
which was considered to represent an acceptable level of accuracy for diets. Diet preparations were
prepared homogenously.

Duration of treatment / exposure:

At least 90 days.

Frequency of treatment:

Once daily at 300 g/animal/day

Dose / conc.:

1 000 ppm

Remarks:

Basis: nominal in diet

Dose / conc.:

3 500 ppm

Remarks:

Basis: nominal in diet

Dose / conc.:

8 000 ppm

Remarks:

Basis: nominal in diet

No. of animals per sex per dose:

4

Control animals:

yes, plain diet

Details on study design:

- Dose selection rationale: Dose levels were based on the results of a 28-day dietary toxicity study with Beagle dogs and on a palatability study with Beagle dogs. In the palatability study, one male and one female received the test substance at 5000 or 10.000 ppm in the diet for one week per concentration. At 10.000 ppm, food intake was notably reduced (with approximately 60%). At 5000 ppm, food intake levels recovered to normal levels after an initial reduction. No changes in body weight or clinical signs were noted that were considered to be an effect of treatment with the test substance. In the 28-day study, two Beagle dogs per sex received the test substance at 1000, 3500 or 8000 ppm in diet. The highest dose of 8000 ppm was selected based on the results of the palatability study. At 8 000 ppm, one female showed a reduced food intake during most of the treatment period, but variations in food intake were also noted during the pretest phase. This same female showed a reddish discolouration of the duodenum, which would be in line with the irritating properties of the test substance. Body weights appeared unaffected by treatment and no clinical signs of toxicity were observed. Haematology and clinical biochemistry revealed no toxicologically significant changes. Based on the significantly reduced food intake observed at 10.000 ppm in the palatability study, it was decided in consultation with the sponsor to select 8000 ppm as the highest dose level for the 90-day main study.

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes, mortality
- Time schedule: At least twice daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: At least once daily

BODY WEIGHT: Yes
- Time schedule for examinations: Weekly during pretest and treatment

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Daily during pretest and treatment.

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No
- Time schedule for examinations:

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: during pretest and week 13
- Dose groups that were examined: All animals

HAEMATOLOGY: Yes
- Time schedule for collection of blood: during pretest and week 13
- Anaesthetic used for blood collection: No
- Animals fasted: Yes
- How many animals: all animals
- Parameters examined: White blood cells, Red blood cells, Haemoglobin, Haematocrit, Mean corpuscular volume, Mean corpuscular haemoglobin, Mean corpuscular haemoglobin concentration, Platelets, Red blood cell distribution width, Differential leucocyte count (Neutrophils, Lymphocytes, Monocytes, Eosinophils, Basophils), Reticulocytes, Prothrombin time, Activated Partial thromboplastin time.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: during pretest and week 13
- Animals fasted: Yes
- How many animals: all animals
- Parameters examined: Aspartate aminotransferase, Alanine aminotransferase, Alkaline phosphatase, Gamma glutamyl transferase, Lactate dehydrogenase, Glutamate dehydrogenase, Total Bilirubin, Glucose, Creatinine, Urea, Total Protein, albumin, Total globulin, Albumin Globulin ratio, Cholesterol, Triglycerides, Phospholipids, Sodium, Potassium, Chloride, Calcium, Inorganic Phosphate.

URINALYSIS: Yes
- Time schedule for collection of urine: during pretest and week 13
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes
- Parameters examined: Volume, Specific gravity, Clarity, Colour, pH, Blood, Leucocytes, Bilirubin, Urobilinogen, Protein, Ketones, Glucose, Nitrite Sediment (Leucocytes, Erythrocytes, Casts, Epithelial cells, Crystals, Bacteria, Other)

NEUROBEHAVIOURAL EXAMINATION: No

Sacrifice and pathology:

Organ weights: Adrenals, Brain, Epididymides, Heart, Kidneys, Liver, Ovaries, Pituitary gland, Prost
ate, Spleen, Testes, Thymus, Thyroid with parathyroids, Uterus

GROSS PATHOLOGY: Yes (see table)

HISTOPATHOLOGY: Yes (see table)

Statistics:

The following statistical methods were used to analyse the data:
- If the variables could be assumed to follow a normal distribution, the Dunnett-test (many-to-one ttest) based on a pooled variance estimate was applied for the comparison of the treated groups and the control groups for each sex.
- The Steel-test (many-to-one rank test) was applied when the data could not be assumed to follow a normal distribution.
- The exact Fisher-test was applied to frequency data.
All tests were two-sided and in all cases p < 0.05 was accepted as the lowest level of significance.
Group means were calculated for continuous data and medians were calculated for discrete data (scores) in the summary tables.

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, treatment-related

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

no effects observed

Haematological findings:

no effects observed

Clinical biochemistry findings:

effects observed, treatment-related

Urinalysis findings:

no effects observed

Behaviour (functional findings):

not examined

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

effects observed, treatment-related

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Details on results:

CLINICAL SIGNS AND MORTALITY
Faeces with red particles were recorded for one male at 8000 ppm (no. 13). Two males at 3500 ppm (nos. 9 and 11) showed a lean appearance from week 2 and 6 onwards, respectively.

BODY WEIGHT AND WEIGHT GAIN
A reduced body weight gain/weight loss was noted for 3 out of 4 males at 3500 ppm and all males at 8000 ppm, throughout most of the treatment period. Slight weight loss primarily occurred in the first five weeks of treatment. A slightly lower body weight gain/slight weight loss throughout treatment was also recorded for one female at 3500 ppm (no. 26) and one female at 8000 ppm (no. 30). Body weights of males at 3500 and 8000 ppm were already slightly lower than control levels during the pretest phase. Body weights and body weight gain of other treated groups remained in the same range as controls over the study period.

FOOD CONSUMPTION AND COMPOUND INTAKE
Food consumption of males at 8000 ppm was slightly reduced during most of the treatment period (achieving a level of statistical significance on most occasions during the first 6 weeks), but slightly recovered towards control levels as treatment progressed. Food intake among females at 8000 ppm appeared slightly reduced in the first week only, achieving a level of statistical significance. No further differences in food intake of these females could be discerned. A significant variation in individual food intake values was noted in primarily females. As these variations in food intake were also apparent among control animals, it was considered that this was due to palatability of the pelleted basal diet (derived from a ground, extruded diet).

CLINICAL CHEMISTRY
The following statistically significant changes in clinical biochemistry parameters distinguished treated animals from control animals:
- Decreased total protein levels in males at 8000 ppm,
- Decreased albumin levels in males at 3500 and 8000 ppm,
- Decreased potassium levels in males and females at 8000 ppm,
- Decreased calcium levels in males at 3500 and 8000 ppm,
- Increased sodium and chloride levels in females at 8000 ppm (not statistically significant for sodium levels),
- Increased inorganic phosphate levels in females at 8000 ppm.

GROSS PATHOLOGY
Necropsy findings distinguishing treated dogs from control dogs were essentially confined to the stomach and included:
- Irregular surface of the pylorus (2/4 females at 1000 ppm and 2/4 females at 8000 ppm), or cardia (1 /4 females at 8000 ppm),
- Reddish foci on the pylorus and/or cardia (1/4 females at 1000 ppm and 1/4 females at 8000 ppm). In addition, one male at 3500 ppm (no. 9) showed an emaciated appearance along with a reduced size of the prostate and thymus.

HISTOPATHOLOGY: NON-NEOPLASTIC
In the stomach, congestion was seen in one female at 1000 and 8000 ppm, correlating to the reddish foci at necropsy. In two females at 1000 and 8000 ppm, the lymphoid follicles in the stomach correlated to the irregular surface recorded at necropsy. These microscopic findings were however within background pathology, and hence were not clearly related to the necropsy findings. Minor grades of diffuse hypertrophy of the zona fasciculata of the adrenals were recorded in 3/4 males at 1000 ppm) and 3/4 males at 3500 ppm, and in 3/4 males and 3/4 females at 8000 ppm (minimal-slight). This finding at these grades is a not uncommon non-specific response to stress in dogs and was considered to be a secondary physiological adaptive change of no toxicological significance.

Dose descriptor:

NOAEL

Effect level:

8 000 ppm

Sex:

male/female

Basis for effect level:

other: No toxicologically significant effects observed

Key result

Dose descriptor:

NOAEL

Effect level:

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

Based on:

test mat.

Sex:

male

Basis for effect level:

other: No toxicologically significant effects observed

Key result

Dose descriptor:

NOAEL

Effect level:

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

Based on:

test mat.

Sex:

female

Basis for effect level:

other: No toxicologically significant effects observed

Critical effects observed:

not specified

Mean test article intake over the study period was as follows:

group	Nominal dietary inclusion level [ppm]	Average intake [mg test substance/ kg body weight/ day] (range indicated within brackets)
		males	females
2	1000	30 (27-35)	36 (30-43)
3	3500	133 (128-139)	114 (107-125)
4	8000	260 (229-275)	255 (229-278)

Conclusions:

NOAEL: 8000 ppm (260 mg/kg/day for males, or 255 mg/kg/day for females).

Executive summary:

90-Day oral toxicity study with p-TSA by dietary administration in male and female Beagle dogs.

Guidelines

The study was based on the following guidelines:

- OECD 409, “Repeated Dose 90-day Oral Toxicity Study in Non-Rodents”, 1998.

- EC Directive 87/302/EEC, B.27: “90-days repeated Oral Dose Study using Non-rodent species”, 1988.

- OPPTS 870.3150, 90-day oral toxicity in nonrodents. Office of Prevention, Pesticides and Toxic

Substances (7101), EPA 712-C-98-200, August 1998.

Rationale for dose levels

Dose levels were based on the results of a 28-day dietary toxicity study with Beagle dogs and on a palatability study with Beagle dogs. In the palatability study, one male and one female received the test substance at 5000 or 10.000 ppm in the diet for one week per concentration. At 10.000 ppm, food intake was notably reduced (with approximately 60%). At 5000 ppm, food intake levels recovered to normal levels after an initial reduction. No changes in body weight or clinical signs were noted that were considered to be an effect of treatment with the test substance. In the 28-day study, two Beagle dogs per sex received the test substance at 1000, 3500 or 8000 ppm in diet. The highest dose of 8000 ppm was selected based on the results of the palatability study. At 8000 ppm, one female showed a reduced food intake during most of the treatment period, but variations in food intake were also noted during the pretest phase. This same female showed a reddish discolouration of the duodenum, which would be in line with the irritating properties of the test substance. Body weights appeared unaffected by treatment and no clinical signs of toxicity were observed. Haematology and clinical biochemistry revealed no toxicologically significant changes. Based on the significantly reduced food intake observed at 10.000 ppm in the palatability study, it was decided in consultation with the sponsor to select 8000 ppm as the highest dose level for the 90-day main study.

Study outline

Beagle dogs received the test substance by dietary intake for at least 90 days. One control group and three treated groups were tested, each consisting of 4 males and 4 females. Evaluated parameters Chemical analysis of prepared diets was conducted to assess homogeneity and accuracy of preparations.

The following parameters were evaluated: clinical signs (daily), body weight (weekly), food consumption (daily), ophthalmoscopic examination (at pretest and end of treatment), clinical pathology (pretest and end of treatment), macroscopy and organ weights at termination, and histopathology on selected tissues.

Results

Dietary analyses confirmed that diets were prepared accurately and homogenously. Treatment at 3500 and 8000 ppm resulted in a reduced weight gain (or slight weight loss on some occasions), primarily among males. Food intake at 8000 ppm was also reduced, again primarily among males. Blood analyses revealed a number of changes at 3500 and 8000 ppm including lower total protein, albumin, potassium and calcium levels, and increased sodium, chloride and inorganic phosphate levels. In the absence of any signs of specific target organ toxicity, these changes were considered to have occurred secondary to the lower body weight/food intake. Necropsy showed stomach effects in a few animals, consisting of an irregular surface of the pylorus (two females at 1000 and 8000 ppm each), or cardia (one female at 8000 ppm), or reddish foci on the pylorus and/or cardia (one female at 1000 and 8000 ppm each). No treatment-related supportive histopathological changes were observed. Also, since the incidence of these findings was not clearly related to the dose, it was considered that there was no adverse effect on the functional integrity of the stomach.

Faeces with red particles were recorded for one male at 8000 ppm. No correlating macro- or microscopic findings were noted to support this finding. Considering its very incidental nature (on a single day throughout treatment), this was considered not indicative of toxicity. No toxicologically significant changes were noted in any of the remaining parameters investigated in this study (i.e. ophthalmoscopy, haematological investigations, urinalysis, organ weights and histopathology).

Conclusion

In the absence of adverse effects on the gastro-intestinal tract or any other organ system, the lower food intake (and lower body weight gain/weight loss) was considered to be related to palatability of the test diet, rather than being indicative of primary systemic toxicity. Therefore, a No Observed Adverse Effect Level (NOAEL) for p-TSA of 8000 ppm was established (corresponding to 260 and 255 mg p-TSA/kg/day for males and females respectively).

These results were used for read-across to Tosylchloramide Sodium.

Reference 4

Endpoint:

sub-chronic toxicity: oral

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

supporting study

Justification for type of information:

The full read across justification report is attached under "Attached justification".

Summary

Within REACH, the obligation to conduct tests with vertebrate animals should be considered as a last resort, only after exhausting all potential sources of information on the physical and (eco)toxicological properties of chemicals [1]. Article 13 of REACH requires that use must be made whenever possible by alternatives to vertebrate animal tests, through the use of alternative methods as in vitro methods, (Q)SARs or from structurally related substances via grouping or read-across.
Annex XI of REACH offers the option to evaluate specific endpoints by read-across. According to Chapter R.6 (QSARs and grouping of chemicals) of REACH technical guidance documents (TGD), read-across can be applied for “substances whose physico-chemical, toxicological and ecotoxicological properties are likely to be similar or follow a regular pattern as a result of structural similarity”. Application of the read-across concept requires that physico-chemical properties, human health effects and environmental effects or environmental fate may be predicted from data of a reference substance to make a prediction for the endpoint of a target chemical. This avoids the need to test every substance for every endpoint. This report provides the justification to use a cross-reading approach between Toluenesulfonamides (p-Toluenesulfonamide (p-TSA, CAS 70-55-3), o-Toluenesulfonamide (o-TSA, CAS 88-19-7) and o/p-Toluenesulfonamide (o/p-TSA, CAS 1333 07 9)) and is presented according to the requirements stipulated in Chapter R.6 (QSARs and grouping of chemicals) of the REACH TGD for N-chloro-4-methylbenzenesulfonamide sodium (Chloramine-T, CAS 127-65-1).

Reason / purpose for cross-reference:

read-across source

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, treatment-related

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

no effects observed

Haematological findings:

no effects observed

Clinical biochemistry findings:

effects observed, treatment-related

Urinalysis findings:

no effects observed

Behaviour (functional findings):

not examined

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

effects observed, treatment-related

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Details on results:

CLINICAL SIGNS AND MORTALITY
Faeces with red particles were recorded for one male at 8000 ppm (no. 13). Two males at 3500 ppm (nos. 9 and 11) showed a lean appearance from week 2 and 6 onwards, respectively.

BODY WEIGHT AND WEIGHT GAIN
A reduced body weight gain/weight loss was noted for 3 out of 4 males at 3500 ppm and all males at 8000 ppm, throughout most of the treatment period. Slight weight loss primarily occurred in the first five weeks of treatment. A slightly lower body weight gain/slight weight loss throughout treatment was also recorded for one female at 3500 ppm (no. 26) and one female at 8000 ppm (no. 30). Body weights of males at 3500 and 8000 ppm were already slightly lower than control levels during the pretest phase. Body weights and body weight gain of other treated groups remained in the same range as controls over the study period.

FOOD CONSUMPTION AND COMPOUND INTAKE
Food consumption of males at 8000 ppm was slightly reduced during most of the treatment period (achieving a level of statistical significance on most occasions during the first 6 weeks), but slightly recovered towards control levels as treatment progressed. Food intake among females at 8000 ppm appeared slightly reduced in the first week only, achieving a level of statistical significance. No further differences in food intake of these females could be discerned. A significant variation in individual food intake values was noted in primarily females. As these variations in food intake were also apparent among control animals, it was considered that this was due to palatability of the pelleted basal diet (derived from a ground, extruded diet).

CLINICAL CHEMISTRY
The following statistically significant changes in clinical biochemistry parameters distinguished treated animals from control animals:
- Decreased total protein levels in males at 8000 ppm,
- Decreased albumin levels in males at 3500 and 8000 ppm,
- Decreased potassium levels in males and females at 8000 ppm,
- Decreased calcium levels in males at 3500 and 8000 ppm,
- Increased sodium and chloride levels in females at 8000 ppm (not statistically significant for sodium levels),
- Increased inorganic phosphate levels in females at 8000 ppm.

GROSS PATHOLOGY
Necropsy findings distinguishing treated dogs from control dogs were essentially confined to the stomach and included:
- Irregular surface of the pylorus (2/4 females at 1000 ppm and 2/4 females at 8000 ppm), or cardia (1 /4 females at 8000 ppm),
- Reddish foci on the pylorus and/or cardia (1/4 females at 1000 ppm and 1/4 females at 8000 ppm). In addition, one male at 3500 ppm (no. 9) showed an emaciated appearance along with a reduced size of the prostate and thymus.

HISTOPATHOLOGY: NON-NEOPLASTIC
In the stomach, congestion was seen in one female at 1000 and 8000 ppm, correlating to the reddish foci at necropsy. In two females at 1000 and 8000 ppm, the lymphoid follicles in the stomach correlated to the irregular surface recorded at necropsy. These microscopic findings were however within background pathology, and hence were not clearly related to the necropsy findings. Minor grades of diffuse hypertrophy of the zona fasciculata of the adrenals were recorded in 3/4 males at 1000 ppm) and 3/4 males at 3500 ppm, and in 3/4 males and 3/4 females at 8000 ppm (minimal-slight). This finding at these grades is a not uncommon non-specific response to stress in dogs and was considered to be a secondary physiological adaptive change of no toxicological significance.

Key result

Dose descriptor:

NOAEL

Effect level:

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

Based on:

test mat.

Sex:

male

Basis for effect level:

other: No toxicologically significant effects observed.

Key result

Dose descriptor:

NOAEL

Effect level:

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

Based on:

test mat.

Sex:

female

Basis for effect level:

other: No toxicologically significant effects observed.

Dose descriptor:

NOAEL

Effect level:

8 000 ppm

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No toxicologically significant effects observed

Critical effects observed:

not specified

Mean test article intake over the study period was as follows:

group	Nominal dietary inclusion level [ppm]	Average intake [mg test substance/ kg body weight/ day] (range indicated within brackets)
		males	females
2	1000	30 (27-35)	36 (30-43)
3	3500	133 (128-139)	114 (107-125)
4	8000	260 (229-275)	255 (229-278)

Conclusions:

NOAEL: 8000 ppm (260 mg/kg/day for males, or 255 mg/kg/day for females).

Executive summary:

90-Day oral toxicity study with p-TSA by dietary administration in male and female Beagle dogs.

Guidelines

The study was based on the following guidelines:

- OECD 409, “Repeated Dose 90-day Oral Toxicity Study in Non-Rodents”, 1998.

- EC Directive 87/302/EEC, B.27: “90-days repeated Oral Dose Study using Non-rodent species”, 1988.

- OPPTS 870.3150, 90-day oral toxicity in nonrodents. Office of Prevention, Pesticides and Toxic

Substances (7101), EPA 712-C-98-200, August 1998.

Rationale for dose levels

Dose levels were based on the results of a 28-day dietary toxicity study with Beagle dogs and on a palatability study with Beagle dogs. In the palatability study, one male and one female received the test substance at 5000 or 10.000 ppm in the diet for one week per concentration. At 10.000 ppm, food intake was notably reduced (with approximately 60%). At 5000 ppm, food intake levels recovered to normal levels after an initial reduction. No changes in body weight or clinical signs were noted that were considered to be an effect of treatment with the test substance. In the 28-day study, two Beagle dogs per sex received the test substance at 1000, 3500 or 8000 ppm in diet. The highest dose of 8000 ppm was selected based on the results of the palatability study. At 8000 ppm, one female showed a reduced food intake during most of the treatment period, but variations in food intake were also noted during the pretest phase. This same female showed a reddish discolouration of the duodenum, which would be in line with the irritating properties of the test substance. Body weights appeared unaffected by treatment and no clinical signs of toxicity were observed. Haematology and clinical biochemistry revealed no toxicologically significant changes. Based on the significantly reduced food intake observed at 10.000 ppm in the palatability study, it was decided in consultation with the sponsor to select 8000 ppm as the highest dose level for the 90-day main study.

Study outline

Beagle dogs received the test substance by dietary intake for at least 90 days. One control group and three treated groups were tested, each consisting of 4 males and 4 females. Evaluated parameters Chemical analysis of prepared diets was conducted to assess homogeneity and accuracy of preparations.

The following parameters were evaluated: clinical signs (daily), body weight (weekly), food consumption (daily), ophthalmoscopic examination (at pretest and end of treatment), clinical pathology (pretest and end of treatment), macroscopy and organ weights at termination, and histopathology on selected tissues.

Results

Dietary analyses confirmed that diets were prepared accurately and homogenously. Treatment at 3500 and 8000 ppm resulted in a reduced weight gain (or slight weight loss on some occasions), primarily among males. Food intake at 8000 ppm was also reduced, again primarily among males. Blood analyses revealed a number of changes at 3500 and 8000 ppm including lower total protein, albumin, potassium and calcium levels, and increased sodium, chloride and inorganic phosphate levels. In the absence of any signs of specific target organ toxicity, these changes were considered to have occurred secondary to the lower body weight/food intake. Necropsy showed stomach effects in a few animals, consisting of an irregular surface of the pylorus (two females at 1000 and 8000 ppm each), or cardia (one female at 8000 ppm), or reddish foci on the pylorus and/or cardia (one female at 1000 and 8000 ppm each). No treatment-related supportive histopathological changes were observed. Also, since the incidence of these findings was not clearly related to the dose, it was considered that there was no adverse effect on the functional integrity of the stomach.

Faeces with red particles were recorded for one male at 8000 ppm. No correlating macro- or microscopic findings were noted to support this finding. Considering its very incidental nature (on a single day throughout treatment), this was considered not indicative of toxicity. No toxicologically significant changes were noted in any of the remaining parameters investigated in this study (i.e. ophthalmoscopy, haematological investigations, urinalysis, organ weights and histopathology).

Conclusion

In the absence of adverse effects on the gastro-intestinal tract or any other organ system, the lower food intake (and lower body weight gain/weight loss) was considered to be related to palatability of the test diet, rather than being indicative of primary systemic toxicity. Therefore, a No Observed Adverse Effect Level (NOAEL) for p-TSA of 8000 ppm was established (corresponding to 260 and 255 mg p-TSA/kg/day for males and females respectively).

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

NOAEL

65 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Quality of whole database:

High.

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

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Principles of method if other than guideline:

Method: other: Standard CIVO procedure, TNO, Zeist, The Netherlands

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Sex:

male/female

Type of coverage:

open

Vehicle:

water

Details on exposure:

Route of Administration: dermal

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

21 days

Frequency of treatment:

Five times a week for 6-8 hours exposure

Remarks:

Doses / Concentrations:
25, 75 and 225 mg/kg bw/day
Basis:
nominal per unit body weight

No. of animals per sex per dose:

2

Control animals:

yes

Details on study design:

Post-exposure period: no

Key result

Dose descriptor:

NOEL

Effect level:

25 mg/kg bw/day

Based on:

not specified

Sex:

male/female

Remarks on result:

other: Effects on skin

Key result

Dose descriptor:

NOEL

Effect level:

225 mg/kg bw/day (nominal)

Based on:

not specified

Sex:

male

Basis for effect level:

clinical signs

Remarks on result:

other: No systemic effects observed up to 225 mg/kg bw/day.

Dose descriptor:

LOEL

Effect level:

75 mg/kg bw/day

Based on:

not specified

Sex:

male/female

Remarks on result:

other: Effects on skin

Critical effects observed:

not specified

NOAEL (NOEL), LOAEL (LOEL): = 25 mg/kg bw, 75 mg/kg bw

ACTUAL DOSE RECEIVED BY DOSE LEVEL BY SEX
- Time of death: no mortality or abnormalities observed.
- Number of deaths at each dose: -

TOXIC RESPONSE/EFFECTS BY DOSE LEVEL: 
- Mortality and time to death: after 21 days 
- Clinical signs: at 225 mg/kg bw distinct skin lesions, scaliness, coriaceousness, haemorrhages, fissuring and sloughing was found. A slight scaliness and very light coriaceousness was observed at 75 mg/kg bw. All experimental animals showed a yellowish discolouration of the fur. No systemic effects observed up to 225 mg/kg bw/day.

- Body weight gain: not significantly adversely affected
- Food/water consumption: not significantly adversely affected.
- Clinical chemistry: -
- Haematology: not significantly adversely affected.
- Urinalysis: not significantly adversely affected.
- Organ weights: not significantly adversely affected.
- Gross pathology: No relevant pathology was found. 
- Histopathology: No relevant abnormalities found. 
- Other: - 

STATISTICAL RESULTS: not described

Conclusions:

The No Observed Effect Level (NOEL) for systemic effects is 225 mg/kg bw/day for effects on the skin the NOEL is 25 mg/kg bw/day.

Executive summary:

A sub acute 21 -day dermal toxicity study was performed at TNO, The Netherlands with albino rabbits. Repeated dermal application of Tosylchloramide sodium, trihydrate at a dose of 225 mg/kg bw/day resulted in distinct skin lesions consiting of scaliness, coriaceousness, haemorrhages, fissuring and sloughing. A slight scaliness and very slight coriaceousness of the exposed skin was seen in the animals of the group treated with 75 mg/kg bw/day. No distinct skin lesions were observed in the lowest dose group 25 mg/kg bw/day. All experimental animals showed a yellowish discolouration of the fur as a result of the application of Tosylchloramide sodium, trihydrate. General appearmce and behaviour, growth, food and water consumption, blood and urine composition, and organ weights were not adversely affected by the repeated dermal application Tosylchloramide sodium, trihydrate. Histologically, distinct changes of the treated skin were found in the two highest dose groups.

The No Observed Effect Level (NOEL) for systemic effects is 225 mg/kg bw/day for effects on the skin the NOEL is 25 mg/kg bw/day.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

NOAEL

225 mg/kg bw/day

Study duration:

subacute

Species:

rabbit

Quality of whole database:

NOEL for effects on the skin is 225 mg/kg bw/day.

Repeated dose toxicity: dermal - local effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Principles of method if other than guideline:

Method: other: Standard CIVO procedure, TNO, Zeist, The Netherlands

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Sex:

male/female

Type of coverage:

open

Vehicle:

water

Details on exposure:

Route of Administration: dermal

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

21 days

Frequency of treatment:

Five times a week for 6-8 hours exposure

Remarks:

Doses / Concentrations:
25, 75 and 225 mg/kg bw/day
Basis:
nominal per unit body weight

No. of animals per sex per dose:

2

Control animals:

yes

Details on study design:

Post-exposure period: no

Key result

Dose descriptor:

NOEL

Effect level:

25 mg/kg bw/day

Based on:

not specified

Sex:

male/female

Remarks on result:

other: Effects on skin

Key result

Dose descriptor:

NOEL

Effect level:

225 mg/kg bw/day (nominal)

Based on:

not specified

Sex:

male

Basis for effect level:

clinical signs

Remarks on result:

other: No systemic effects observed up to 225 mg/kg bw/day.

Dose descriptor:

LOEL

Effect level:

75 mg/kg bw/day

Based on:

not specified

Sex:

male/female

Remarks on result:

other: Effects on skin

Critical effects observed:

not specified

NOAEL (NOEL), LOAEL (LOEL): = 25 mg/kg bw, 75 mg/kg bw

ACTUAL DOSE RECEIVED BY DOSE LEVEL BY SEX
- Time of death: no mortality or abnormalities observed.
- Number of deaths at each dose: -

TOXIC RESPONSE/EFFECTS BY DOSE LEVEL: 
- Mortality and time to death: after 21 days 
- Clinical signs: at 225 mg/kg bw distinct skin lesions, scaliness, coriaceousness, haemorrhages, fissuring and sloughing was found. A slight scaliness and very light coriaceousness was observed at 75 mg/kg bw. All experimental animals showed a yellowish discolouration of the fur. No systemic effects observed up to 225 mg/kg bw/day.

- Body weight gain: not significantly adversely affected
- Food/water consumption: not significantly adversely affected.
- Clinical chemistry: -
- Haematology: not significantly adversely affected.
- Urinalysis: not significantly adversely affected.
- Organ weights: not significantly adversely affected.
- Gross pathology: No relevant pathology was found. 
- Histopathology: No relevant abnormalities found. 
- Other: - 

STATISTICAL RESULTS: not described

Conclusions:

The No Observed Effect Level (NOEL) for systemic effects is 225 mg/kg bw/day for effects on the skin the NOEL is 25 mg/kg bw/day.

Executive summary:

A sub acute 21 -day dermal toxicity study was performed at TNO, The Netherlands with albino rabbits. Repeated dermal application of Tosylchloramide sodium, trihydrate at a dose of 225 mg/kg bw/day resulted in distinct skin lesions consiting of scaliness, coriaceousness, haemorrhages, fissuring and sloughing. A slight scaliness and very slight coriaceousness of the exposed skin was seen in the animals of the group treated with 75 mg/kg bw/day. No distinct skin lesions were observed in the lowest dose group 25 mg/kg bw/day. All experimental animals showed a yellowish discolouration of the fur as a result of the application of Tosylchloramide sodium, trihydrate. General appearmce and behaviour, growth, food and water consumption, blood and urine composition, and organ weights were not adversely affected by the repeated dermal application Tosylchloramide sodium, trihydrate. Histologically, distinct changes of the treated skin were found in the two highest dose groups.

The No Observed Effect Level (NOEL) for systemic effects is 225 mg/kg bw/day for effects on the skin the NOEL is 25 mg/kg bw/day.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Study duration:

subacute

Species:

rabbit

Quality of whole database:

NOEL for effects on the skin is 25 mg/kg bw/day.

Additional information

Repeated dose – by oral route:

Repeated dose toxicity oral-90 day rat with p-TSA (OECDTG 408, Van Otterdijk, 2007)

A repeated dose 90-Day oral toxicity study with by daily gavage of p-Toluenesulfonamide (p-TSA) in the rat was performed according to OECD TG 408. Based on a 14-day oral range finding study, the dose levels for this 90-day oral gavage study were selected to be 0, 1000, 3000 and 10 000 ppm (0, 70, 214 and 738 mg/kg body weight/day for males, and 0, 80, 248 and 795 mg/kg body weight/day for females). The test substance was administered daily for at least 90 days by oral gavage to SPF-bred Wistar Han rats. One control group and three treated groups were tested, each consisting of 10 males and 10 females.

Chemical analyses of diet preparations were conducted to assess accuracy and homogeneity of diet preparations.

The following parameters were evaluated: Clinical signs (daily), functional observations (week 13), body weight and food consumption (weekly), and ophthalmoscopy (pretest and week 13). At termination: clinical pathology, macroscopy, organ weights and histopathology on a selection of tissues.

Homogeneity and accuracy of diet preparations were demonstrated by analyses.

Dietary inclusion levels of 0, 1000, 3000 and 10.000 ppm were equivalent to an average test article intake of 0, 70, 214 and 738 mg/kg body weight/day for males, and 0, 80, 248 and 795 mg/kg body weight/day for females.

 

At 10.000 ppm, a significant and consistent reduction of body weight gain was recorded (up to 21% compared to control animals).

Histopathology revealed a minimal degree of hyperplasia of the urothelium of the urinary bladder in two males at 10.000 ppm.

 No other treatment-related toxicologically significant changes were noted in any of the remaining parameters examined/determined in this study (i.e. clinical appearance, functional observations, food consumption, clinical laboratory investigations, macroscopic examination and organ weights).

 

The observed minimal urethelial hyperplasia in two animals seems to be caused by local irritation by the chronic availability of p-TSA metabolite (p-sulphamonylbenzoic acid) in the urine, following the rapid and complete excretion of p-TSA. Such effect is local and fully reversible when exposure ends. As the hyperplasia is observed in only two animals, is minimal and likely a fully reversible temporal effect upon exposure, is not an adverse effect persé, the reduction of the body weight gain of 21% in males is considered serious enough to assign the 10000 ppm a LOAEL. Especially as a relation to palatability is not completely clear.

 

LOAEL: 10.000 ppm (738 mg/kg/day for males, or 795 mg/kg/day for females).

NOAEL: 3000 ppm (214 mg/kg/day for males, or 248 mg/kg/day for females).

Repeated dose toxicity oral-90d-dog with p-TSA (OECDTG 409, Van Otterdijk, 2008)

Additionally a 90-day oral repeated dose toxicity study was performed in a second species. p-TSA was dose by dietary administration in groups of four male and four female Beagle dogs at levels of 0, 1000, 3500 and 8000 ppm in the diet, equivalent to an average intake of 0, 30, 133 and 260 mg/kg body weight/day for males, and 0, 36, 114 and 255 mg/kg body weight/day for females.

Results:

Treatment at 3500 and 8000 ppm resulted in a reduced weight gain (or slight weight loss on some occasions), primarily among males. Food intake at 8000 ppm was also reduced, again primarily

among males. Blood analyses revealed a number of changes at 3500 and 8000 ppm including lower total protein, albumin, potassium and calcium levels, and increased sodium, chloride and inorganic phosphate levels.

In the absence of any signs of specific target organ toxicity, these changes were considered to have occurred secondary to the lower body weight/food intake.

Necropsy and histopathological evaluations did not show effects indicative of toxicity. No toxicologically significant changes were noted in any of the remaining parameters investigated in this study (i.e.

ophthalmoscopy, haematological investigations, urinalysis, organ weights and histopathology).

In the absence of adverse effects on the gastro-intestinal tract or any other organ system, the lower food intake (and lower body weight gain/weight loss) was considered to be related to palatability of the test diet, rather than being indicative of primary systemic toxicity.

NOAEL: 8000 ppm (corresponding to 260 and 255 mg p-TSA/kg/day for males and females respectively).

Additional relevant data from reproduction studies reported in this dossier:

Toxicity to reproduction 2 -generation oral- rat with p-TSA (OECDTG 416, Beekhuizen, 2007)

Animals were dosed 0, 1000, 3000 and 10,000 ppm in the diet. F0-parents were dose for 10 weeks prior to mating and continuing during mating until euthanasia. The F1-generation was potentially exposed to the test substance in utero, through nursing during lactation and directly following weaning, and then treated for a minimum of 10 weeks prior to mating and continuing until euthanasia. Only observed effects at 3000 ppm consisted of decreased and body weight gain. In F0-parents, terminal body weights at necropsy were decreased (92% of control) which resulted in relative organ weight changes for the brain and kidneys (105% and 110% of control, respectively). In the F1-generation, terminal body weights at necropsy were also decreased (94% of control) which resulted in relative organ weight changes for the brain (107% of control).

Consequently, the NOAEL was set at 1000 ppm (corresponding to 52-78 mg/kg bw/day for males (average 65) and 75-161mg/kg bw/day for females) based on decreased body weights and body weight gain, and due to this organ weight changes in both F0 and F1 parental animals.

Combined Repeat Dose and Reproductive/Developmental Toxicity Screening Test OECD 422 oral-rat with p-TSA, 1994

Additional information is available from testing performed for OECD HPV evaluation by Japan, and reported in SIDS dossier on p-TSA.

OECD 422, Combined Repeat Dose and Reproductive/Developmental Toxicity Screening Test in rat by gavage under GLP.

Dose levels: 0 (vehicle, 5% Arabic gum), 120, 300, 750 mg/kg/day, with 13 animals, 3 per dose group per sex.

Dosing period: Females, from 14 days before mating to day 3 of lactation. Males 42 days.

750 mg group: decreased BW and food consumption. Also haematuria was observed in 4/13 animals within the first 3 days of dosing.

Decrease in the thymus weight in females (acceleration of involution), and Slight, but statistically significant, increases in relative kidney and testicular weights were found in the males, and in relative kidney and liver weights in females likely related to the decreased BW.

Dark-coloured liver in 6/13 males at gross pathological evaluation Increased WBC in males with increase in the proportion of neutrophils. Also in males increased BUN, ASAT, ALAT, chloride and potassium levels. Histopathology showed thickened urinary bladder epithelium in 11/13 males, and 7/13 females.

300 mg: Decreased BW and food consumption females during gestation and lactation Decrease in the thymus weight in females (acceleration of involution) Increased WBC in males with increase in the proportion of neutrophils, and increased BUN, ASAT and chloride.

Histopathology showed thickened urinary bladder epithelium in 11/13 males, and 12/13 females.

120 mg: Only reportable effect observed is thickened urinary bladder epithelium in 6/13 males and 1/13 females at histopathological examination. Although this effect does not seem to show a clear dose response relation, it was not observed in any of the control animals. Therefore the NOEL was set at < 120 mg/kg bw/day.

Information available on benzenesulfonamides supports the common mechanism of action for toxicity to urinary tract observed in repeated dose studies with these compounds. The effect is indirect mediated by the appearence of calcium phosphate calculi caused by an indueced alkaline urine.

Developmental toxicity study oral-rabbit with p-TSA (OECDTG 414 van Tuyl, 2008)

Developmental toxicity was evaluated in an OECD 414 study in in which rabbits were dosed by dietary exposure to 0, 1000, 3000 and 11000 ppm (equivalent to 0, 41, 113 and 367 mg/kg body weight/day respectively) from days 7 to 29 post-coitum. Based on the effects on body weight, food and water consumption, the maternal NOAEL for p-TSA was established as being 3000 ppm (114 mg/kg body weight/day), although some transient effects on body weight were noted at 3000 ppm.

Repeated dose – by inhalation:

No study available.

Repeated dose – by dermal route:

Repeated dose toxicity dermal-21day rabbit wih Halamid (pre OECD , Huismans and Van der Meulen, 1971)

A sub acute 21 -day dermal toxicity study was performed at TNO, The Netherlands with albino rabbits. Repeated dermal application of Halamid at a dose of 225 mg/kg bw/day resulted in distinct skin lesions consiting of scaliness, coriaceousness, haemorrhages, fissuring and sloughing. A slight scaliness and very slight coriaceousness of the exposed skin was seen in the animals of the group treated with 75 mg/kg bw/day. No distinct skin lesions were observed in the lowest dose group 25 mg/kg bw/day. All experimental animals showed a yellowish discolouration of the fur as a result of the application of Halamid. General appearmce and behaviour, growth, food and water consumption, blood and urine composition, and organ weights were not adversely affected by the repeated dermal application Halamid. Histologically, distinct changes of the treated skin were found in the two highest dose groups.

The No Observed Effect Level (NOEL) for systemic effects is 225 mg/kg bw/day for effects on the skin the NOEL is 25 mg/kg bw/day.

Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:

Of the two available highest quality reports from studies involving the longest duration (90-days) one in rat and one in dog, the rat studies resulted to the lowest NOAEL of 3000 ppm (concentration in diet involves automatic correction for allometric scaling) equivalent to 214 and 248 mg p-TSA/kg/day for males and females respectively. However, a 2-generation study of similar quality indicates some effects of reduced body weight especially in pregnant and lactating animals from 3000 ppm, resulting to a NOAEL of 1000 ppm, equivalent to 52 -78 (average 65) mg/kg bw/day for males and 75-161 mg/kg bw/day for females. Also a recent developmental study mentions transient effects at 3000 ppm, and no effects at 1000 ppm. Repeated dose toxicity: via oral route - systemic effects (target organ) urogenital: urinary bladder

These results were used for read-across to Tosylchloramide Sodium.

Justification for classification or non-classification

Available studies do not indicate a need for classification for STOT-RE. p-TSA is of limited toxicity at levels of 100 mg/kg bw/day and below.

Most relevant:

Toxicty to reproduction study oral in rats 2 -generation study in rats (OECDTG 416):

NOAEL 65 mg/kgbw/d (based on 1000 ppm in diet) due to transient effects on body weights at 3000 ppm (read-across from p-TSA).

These results were used for read-across to Tosylchloramide sodium.

Tosylchloramide sodium does not have to be classified for specific target organ toxicity following repeated exposure in accordance with the criteria outlined in Annex I of the CLP Regulation (1272/2008/EC).