Registration Dossier

Toxicological information

Repeated dose toxicity: oral

Currently viewing:

Administrative data

Endpoint:
repeated dose toxicity: oral, other
Remarks:
combined repeated dose and reproduction / developmental screening
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2003
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: GLP study according to guideline. Although only summary available it has been peer internationally reviewed within OECD and assigned reliability 1.
Cross-reference
Reason / purpose for cross-reference:
reference to same study

Data source

Reference
Reference Type:
publication
Title:
Unnamed
Year:
2003

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Deviations:
no
GLP compliance:
yes
Limit test:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
Tosyl chloride
EC Number:
202-684-8
EC Name:
Tosyl chloride
Cas Number:
98-59-9
Molecular formula:
C7H7ClO2S
IUPAC Name:
4-methylbenzene-1-sulfonyl chloride
Details on test material:
4-Methylbenzenesulfonyl chloride, purity = 99.7 %,
Fluka, Lot No. - 422308/1

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
- male/female
- Age of animals at study: 9 weeks old for males and females
- Weight at study repeated dose toxicity: 325.8 - 363.1 g for males and 198.5 - 229.3 g for females
- Number of test animals: 60 animals for each sex

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
not specified
Details on oral exposure:
According to the preliminary test (Report No. P104), dose level was set to 0, 150, 350 and 750 mg/kg/day.
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
Exposure period : 34, 36 - 45, and 51 days for male, copulated female and not copulated female animals, respectively.
Frequency of treatment:
Daily
Doses / concentrations
Remarks:
Doses / Concentrations:
0, 150, 350 and 750 mg/kg/day
Basis:

No. of animals per sex per dose:
12 animals/sex/dose, plus 6 in 14-day recovery group/sex for control and high dose
Control animals:
yes, concurrent vehicle
Details on study design:
Including 14 day recovery groups for control and high dose

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily; mortality: twice/day
- Cage side observations checked in table [No.?] were included.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Weekly

BODY WEIGHT: Yes
- Time schedule for examinations: once a week and just before the necropsy, but in case of pregnant females, it was measured on the day 0, 7, 14, 20 of the gestation period, date of delivery, and 4 days of the lactation day.

FOOD CONSUMPTION:
- Food consumption: Yes: once a week except mating period

WATER CONSUMPTION: No

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes / No / No data
- Time schedule for collection of blood:
- Anaesthetic used for blood collection: Yes (identity) / No / No data
- Animals fasted: Yes / No / No data
- How many animals:
- Parameters checked in table [No.?] were examined.

HAEMATOLOGY and CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at moment of necropsy
- Animals fasted: Yes
- How many animals: randomly selected 5 male and female rats from each test group
- Parameters examined:
Haematocrit, hemoglobin concentration, erythrocyte count, total and different leucocyte count, platelet count, prothrombin time, and active partial thromboplastin time.
Biochemical test: sodium, potassium, chloride, glucose, total cholesterol, blood urea nitrogen, creatinine, total protein, albumin, alanine aminotransferase, aspartate aminotransferase, and total bilirubin.

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: before necropsy and during lactation for males and females, respectively
- Dose groups that were examined: 5 animals were randomly selected from each test group
- Battery of functions tested: Behaviour, grip strength, prayer reflex test and corneal reflex
Sacrifice and pathology:
GROSS PATHOLOGY: Yes

ORGAN WEIGHT: testes, epididymider (all males) liver, kidney, adrenals, thymus, spleen, brain and heart (5 male and female rats from each test group).

HISTOPATHOLOGY: Yes
22 tissues were fixed to perform histopathological evaluations including: testes, epididymides, ovaries, accessory sex organs for all animals, brain (including cerebrum, cerebellum and pons), spinal cord, stomach, small and large intestines (including Peyer’s patches), liver, kidneys, adrenals, spleen, heart, thymus, thyroid, trachea, lungs, uterus, urinary bladder, lymph nodes (cervical mesenteric), peripheral nerve (sciatic or tibial), and bone marrow.
Statistics:
Statistical decision tree, but in case of recovery group, either two-side Student’s t-test or two-side Aspin-Welch t-test was used. In case of categorical data, two-sided Fisher’s exact test was used.

Results and discussion

Results of examinations

Clinical signs:
effects observed, treatment-related
Mortality:
mortality observed, treatment-related
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:
not examined
Haematological findings:
effects observed, treatment-related
Clinical biochemistry findings:
effects observed, treatment-related
Urinalysis findings:
not examined
Behaviour (functional findings):
no effects observed
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Gross pathological findings:
not specified
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Details on results:
CLINICAL SIGNS AND MORTALITY
No mortality males; in females 350 mg/kg: 2/12 and at 750 mg/kg: 4/18
In the control groups, there were no specific clinical symptoms during test period.
150 mg: Intermittent (blood-like) salivation and staining around mouth (3/18 males, 6/18 females), and in females at delivery: difficult delivery, poor nursing, irregular respiration, uterus introsusception and piloerection.
350 mg: Intermittent (blood-like) salivation and staining around mouth were observed after day 15 and (blood-like) staining around nose (7/12 males, 4/12 females). Iintermittent soft stool and staining around anorectal region were observed for the most male animals and in 2 females. One female showed difficult delivery, lacrimation, and irregular respiration from day 4.
750 mg: soft stool and staining around anorectal region for most males and all females; and 5 cases of loss of hair around tail region were observed. Intermittent (blood-like) salivation and 9 cases of staining around mouth and 9 cases of (blood-like) staining around nose. Soft stool and staining around anorectal region for all animals; some cases of intermittent diarrhea were observed. Some animals with found dead and in dying condition had symptoms such as irregular respiration, crawing position, hypoactivity, and abdominal swelling.
In the 750 mg/kg/day recovery group, salivation, staining around mouth, soft stool and staining around anorectal region were not observed during the recovery period.

BODY WEIGHT AND WEIGHT GAIN
From start of dosing BW gain at 750 mg/kg group a bit lower resulting to a lower BW of 8% in males and 5.5% in females compared to controls during first week of recovery which partly recovered during the second week of the recovery period.

FOOD CONSUMPTION:
Only during the first week food consumption some somewhat lower in the 750 mg group.

HAEMATOLOGY
No major changes were seen compared to control, although the reported that dose related decrease in RBC and HCT and an increase in platelet counts were observed in males (see table)

CLINICAL CHEMISTRY
In males and females BUN is decreased compared to control at 750 mg/kg but not in the recovery groups. Total cholesterol was decreased in the recovery groups. In females chloride is decreased at 750 mg/kg (See table)

NEUROBEHAVIOUR
No significant effects were found.

ORGAN WEIGHTS
No effects were seen in sex-organ weights. Weight of the spleen was increased in treatment groups compared to controls for both males and females, although in females not relative weight. No differences in spleen weights were seen in recovery groups. In males the absolute, but not the relative, weight of the heart was decreased in the 750 mg/kg group. In the female recovery group the weights for adrenal and the brain were increased. (See table)

GROSS PATHOLOGY
No information provided

HISTOPATHOLOGY: NON-NEOPLASTIC
See table.

Effect levels

Key result
Dose descriptor:
LOAEL
Effect level:
150 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Based on salivation and effects of irritation in nongranular stomach in males and females, and difficulty delivery and poor nursing in females.

Target system / organ toxicity

Critical effects observed:
not specified

Any other information on results incl. tables

Hematology of males (group mean) 

 Group 

 WBC (103/mm3) 

 RBC (106/mm3) 

 HGB (g/dl) 

 HCT (%) 

 PLT (103/mm3) 

 C 

11.2

8

15.4

43.2

910.6

 T1 

13.6

7.7

15.3

41.7

999.2

 T2 

13.7

7.3

14.7

40.3

1,092

 T3 

13.1

7.3

14.7

41

1,100

Recovery (group mean) 

 C 

13.2

7.8

15.1

41.4

917

 T3 

13.7

7.8

15.4

42.9

915.2

 

 

Clinical Biochemistry of males (group mean)

 Treatment 

 TP 

 ALB 

 AST 

 ALT 

 BUN 

 CREA 

 TCHO 

 GLU 

 Na 

 K 

 Cl 

 (mg/kg/day) 

 g/dL 

 g/dL 

 IU/L 

 IU/L 

 mg/dL 

 mg/dL 

 mg/dL 

 mg/dL 

 mmol/L 

 mmol/L 

 mmol/L 

 0 

6

2.3

148.3

46.5

11.3

0.6

98.8

69

147.2

4.6

104

 150 

5.8

2.2

132.5

28.6

10.7

0.5

114.2

64.4

143.8

4.6

102

 350 

5.8

2.3

122.8

31.3

9.6

0.5

102.8

66

145.4

4.9

102.4

 750 

5.6

2.2

127.2

41.3

7.8

0.5

102.8

58.6

145.6

4.4

102.6

Recovery

 0 

5.8

2.3

103.9

32.7

11.7

0.6

116

67.2

143.2

4.7

104.8

 750 

5

1.9

93

31.4

12.8

0.5

105.2

46.4

128.8

4.1

94.2

Clinical Biochemistry of females (group mean)

 0 

5

2

102.5

59.9

14.1

0.7

105.6

85

140.4

4.1

100.4

 150 

5.1

2.1

101.6

54.1

13.6

0.7

112.2

70.6

144

4.5

102.6

 350 

5.4

2.2

94.3

61.1

13.3

0.7

102.8

73

139.4

4.6

100.8

 750 

5.2

2.1

107.2

63.4

11.4

0.6

121.4

79.6

142.2

4.9

99.6

Recovery

 0 

6.7

3.1

125.4

38.8

12.1

0.6

140.6

96.8

145

4.3

107.2

 750 

6.6

2.9

103.9

30.6

15.3

0.6

128

86.6

144.4

4.2

103.6

 

Dose (mg/kg) 

0

150

350

750

 satellite 

0

750

Absolute (sex) organ weight of males

Testes(g) 

3.04

3.285

3.332

3.203

3.337

3.208

Epididymis(g) 

1.258

1.29

1.37

1.3

1.388

1.335

Liver (g) 

10.698

11.142

10.698

10.01

11.139

10.736

Thymus(g) 

0.355

0.307

0.356

0.32

0.31

0.297

Kidneys(g) 

2.364

2.272

2.502

2.385

2.397

2.474

Adrenals(g) 

0.061

0.064

0.062

0.068

0.053

0.055

Spleen(g) 

0.632

0.725

0.783

0.794

0.844

0.716

Brain(g) 

1.985

1.947

2.022

1.91

2.064

2.018

Heart(g) 

1.325

1.244

1.282

1.182

1.411

1.382

Absolute organ weight of females

Liver (g) 

6.947

6.886

7.298

7.431

6.855

6.621

Kidneys(g) 

1.46

1.451

1.433

1.5

1.523

1.509

Adrenals(g) 

0.071

0.073

0.073

0.072

0.057

0.066

Thymus(g) 

0.106

0.14

0.161

0.096

0.298

0.291

Brain(g) 

1.884

1.885

1.883

1.855

1.742

1.87

Spleen(g) 

0.397

0.419

0.494

0.476

0.477

0.472

Heart(g) 

0.818

0.809

0.853

0.767

0.876

0.829

 

Histopathological findings of males (Group)

 Dose level (mg/kg/day) 

0

150

350

700

 No. of animals examined 

5

5

5

5

 Observation(s) 

 No. of animals observed 

No significant findings 

2

4

4

3

Liver: focal necrosis

 

 

 

 

Minimal 

0

0

0

1

Nonglandular stomach: epithelial proliferation and vacuolation

 

 

 

 

Minimal

0

2

0

0

Moderate

0

3

0

0

Marked 

0

0

5

5

Nonglandular stomach: hyperkeratosis

 

 

 

 

Minimal 

0

3

0

0

Slight 

0

2

5

5

Nonglandular stomach: submucosal edema

 

 

 

 

Minimal 

0

0

0

1

Slight 

0

0

4

4

Moderate

0

3

1

0

Nonglandular stomach: inflammation

 

 

 

 

Slight 

0

3

5

5

No. of animals examined (reproductive organs)

12

12

12

12

Observation(s) 

 No. of animals observed 

No significant findings

10

12

12

10

Testes: atrophy and degeneration of seminiferous tubules

 

 

 

 

Minimal

0

0

0

1

Moderate

2

0

0

0

Marked 

0

0

0

1

Epididymides: oligospermia

 

 

 

 

Minimal

1

0

0

0

Marked 

1

0

0

1

 

Histopathological findings of females (Group)

 Dose level (mg/kg/day) 

0

150

350

700

 No. of animals examined 

5

0

0

0

 Observation(s) 

 No. of animals observed 

No significant findings 

5

1

0

0

Liver: focal necrosis

 

 

 

 

Minimal 

0

0

0

0

Heart: inflammatorycell foci

 

 

 

 

Minimal 

1

0

0

0

Nonglandular stomach: epithelial proliferation and vacuolation

 

 

 

 

Minimal

0

1

1

1

Slight 

0

2

3

3

Marked 

0

0

1

1

Nonglandular stomach: hyperkeratosis

 

 

 

 

Minimal 

0

1

3

1

Slight 

0

0

2

3

Moderate

0

0

0

1

Nonglandular stomach: submucosal edema

 

 

 

 

Slight 

0

1

1

1

Nonglandular stomach: inflammation

 

 

 

 

Minimal 

0

1

2

3

No. of animals examined (reproductive organs)

12

12

10

9

Observation(s) 

 No. of animals observed 

No significant findings

12

12

10

9

Applicant's summary and conclusion

Conclusions:
LOAEL = 150 mg/kg bw based on salivation and effects of irritation in nongranular stomach in males and females at this dose level.
Executive summary:

Tosyl chloride was dose by gavage to groups of 12 Sprague-Dawley rats/sex/dose level at levels of 0, 150, 350 and 750 mg/kg/day for 35 days and 36 - 51 days for male and female rats, respectively. A separate group of 6 animals/sex were added for a 14-day recovery group to the control and the high dose groups. Some clinical signs were observed at the dose level of 150 mg/kg/day for males animals such as intermittent (blood-like) salivation and staining around mouth; and for female animals such as intermittent (blood-like) salivation, staining around mouth, difficulty delivery, poor nursing, and irregular respiration. Moreover, at the dose level of 150 mg/kg/day, the digestive system and nonglanular stomach were also affected.

No effects were observed in the animals of the recovery groups.

Consequently the reporting concludes of a NOAEL < 150 mg/kg bw/day.