Tosyl chloride

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Link to relevant study records

Reference

Reference 1

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

experimental study

Adequacy of study:

supporting 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.

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes

Limit test:

no

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

Route of administration:

oral: gavage

Vehicle:

not specified

Details on mating procedure:

Each male and female rat was selected from the same test group in order to copulate. It spent 14 days in terms of mating. The day after the copulating, mating would be determined through the observation of sperm in a vaginal rinse.

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

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 no treatment

Details on study design:

According to the preliminary test (Report No. P104), dose level was set to 0, 150, 350 and 750 mg/kg/day.

Parental animals: Observations and examinations:

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

Oestrous cyclicity (parental animals):

Not determined.

Sperm parameters (parental animals):

Not determined.

Litter observations:

PARAMETERS EXAMINED
The following parameters were examined in F1 offspring:
- The number of survivors and deaths during delivery
- Body weight and Survival rate: measured on the day of delivery and on the day 4 of lactation.
- Sex ratio: It is determined by anogenital distance, for more than 2 mm and less than 1 mm were male and female, respectively.
- Examination of the external surface of pups: It was observed on the day of delivery and on the day 4 of lactation to determine abnormalities.

Postmortem examinations (parental animals):

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.

Postmortem examinations (offspring):

No infoirmation

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.

Reproductive indices:

- Copulation index
- number of implantations
- number of corpus luteum
- Number of females pregnant

Offspring viability indices:

- Number of life pups
- post natal loss
- pre and post-implantation loss

Clinical signs:

effects observed, treatment-related

Body weight and weight changes:

effects observed, treatment-related

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Reproductive function: oestrous cycle:

not examined

Reproductive function: sperm measures:

not examined

Reproductive performance:

no effects observed

CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
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 FOOD CONSUMPTION (PARENTAL ANIMALS)
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.
Only during the first week food consumption some somewhat lower in the 705 mg group.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
- Pregnancy and delivery: No effects were seen between the groups in duration of pregnancy, number of corpus luteum, number of implantation, pre-implantation loss, post-implantation loss
- Index of copulation, fertility and gestation: There was no significant difference between the control and treatment group in terms of copulation, fertility and gestation index. The sex mis-confirmation for new born pups in this test did not have relationship with test substance since its frequency was low and no dose-correlation. (See Table)

ORGAN WEIGHTS (PARENTAL ANIMALS)
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 (PARENTAL ANIMALS)
No information provided

HISTOPATHOLOGY (PARENTAL ANIMALS)
See table.

Key result

Dose descriptor:

NOAEL

Effect level:

750 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: NOAEL parental: Above the highest tested dose (750 mg/kg/day)

Clinical signs:

no effects observed

Mortality / viability:

no mortality observed

Body weight and weight changes:

no effects observed

Sexual maturation:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

no effects observed

Description (incidence and severity):

Examination of the external surface of pups

Histopathological findings:

not specified

Number of pups born, viability and sex ratio: No significant difference was observed between the treatment groups and the control group at the time of the delivery and on the day 4 of the lactation.

Reproductive effects observed:

not specified

Mating, fertility, gestation and viability data:

DOSE: (mg/kg)	0	150	350	750
No. of mated males	12	12	12	11
Copulation index (%)	100	100	100	90.9
Fertility index (%)	91.7	100	100	100
No. of mated females	12	12	12	11
Copulation index (%)	100	100	100	90.9
Fertility index (%)	91.7	100	100	100
Gestation index (%)	100	91.7	83.3	90
No. of corpora lutea	14.9	14.5	15.1	14.1
Mean±S.D	1.1	2.5	1.7	0.9
No. of implantations	14.4	12.7	12.7	13.3
Mean±S.D	1	4.2	2.8	1
Mean % preimplantation loss	3.5	15.5	14.3	5.4
No. of embryo/fetal death	2	1.7	2	2.3
No. of live pups born	12.4	12	10.7	11
Mean±S.D	0.9	2.5	3.3	3.2
Mean pregnancy period (day)	21.7	21.8	21.8	21.9
Viability at birth pp	95.4	92.1	93.9	92.3
Viability at LD 4	99.2	97.8	88.3	99
Body weights of pups (g)
Male (at birth)	6.25	5.99	6.37	5.98
Day 4	8.57	8.56	9.71	8.7
Female (at birth)	5.75	5.62	5.73	5.59
Day 4	8.03	7.98	8.59	8.21

 

 

Organ weights:

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

 

Histopathology:

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
Nongrandular stomach: epithelial proliferation and vacuolation
Minimal	0	2	0	0
Moderate	0	3	0	0
Marked	0	0	5	5
Nongrandular stomach: hyperkeratosis
Minimal	0	3	0	0
Slight	0	2	5	5
Nongrandular stomach: submucosal edema
Minimal	0	0	0	1
Slight	0	0	4	4
Moderate	0	3	1	0
Nongrandular 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
Nongrandular stomach: epithelial proliferation and vacuolation
Minimal	0	1	1	1
Slight	0	2	3	3
Marked	0	0	1	1
Nongrandular stomach: hyperkeratosis
Minimal	0	1	3	1
Slight	0	0	2	3
Moderate	0	0	0	1
Nongrandular stomach: submucosal edema
Slight	0	1	1	1
Nongrandular 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

 

Conclusions:

There were no reproductive effects seen at 750 mg/kg, the highest dose tested

Executive summary:

Tosyl chloride has been evaluated in a Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test according to OECD 422 and in compliance to GLP.

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.

Results:

Parental toxicity: No mortality males; in females 350 mg/kg: 2/12 and at 750 mg/kg: 4/18.

Some clinical signs were observed at the dose level of 150 mg/kg/day in both male and females such as intermittent (blood-like) salivation and staining around mouth. Effects were more severe at 350 mg/kg and at 750 mg/kg reported clinical signs were 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. A decreased BW gain was observed in the 750 mg group resulting to a lower BW that was maximal 8% lower in males and 5.5% in females compared to controls. Food consumption was a bit lower during the first week in the 750 mg group. Histopathology showed dose related effects of epithelial proliferation and inflammation of the forestomach already visible in animals treated at 150 mg.

There were no further major changes observed in other parameters (haematology, biochemistry, neurobehavioral functions, organ weights)

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.

 

Reproductive parameters were not affected.No effects were seen between the groups in duration of pregnancy, number of corpus luteum, number of implantation, pre-implantation loss, post-implantation loss. There was no significant difference between the control and treatment group in terms of copulation, fertility and gestation index.

Among the clinical signs was remarked for the 150 mg/kg treated group only that thefemale animals additionally showed difficulty in delivery, poor nursing, and irregular respiration. However, as this is not remarked for the higher dose groups, and no effects are seen in any of the reproductive parameters, these likely represents an accidental observation.

 

Consequently the NOAEL for fertility was established at 750 mg/kg bw/day, the highest dose tested.

Reference 2

Endpoint:

extended one-generation reproductive toxicity - basic test design (Cohorts 1A, and 1B without extension)

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

the extended one-generation reproductive toxicity study does not need to be conducted because there are no results from available repeated dose toxicity studies that indicate adverse effects on reproductive organs or tissues, or reveal other concerns in relation with reproductive toxicity

other:

Justification for type of information:

Further testing of Tosyl chloride for reproductive toxicity is not indicated on the basis of low exposures, no effects seen on reproductive parameters seen in an OECD 422 screening at highest dose levels tested leading to mortality.

Reproductive effects observed:

not specified

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

750 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

Study of high validity

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

Tosyl chloride has been evaluated in a Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test according to OECD 422 and in compliance to GLP.

 

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.

Results: Parental toxicity: No mortality males; in females 350 mg/kg: 2/12 and at 750 mg/kg: 4/18.

Some clinical signs were observed at the dose level of 150 mg/kg/day in both male and females such as intermittent (blood-like) salivation and staining around mouth. Effects were more severe at 350 mg/kg and at 750 mg/kg reported clinical signs were 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. A decreased BW gain was observed in the 750 mg group resulting to a lower BW that was maximal 8% lower in males and 5.5% in females compared to controls. Food consumption was a bit lower during the first week in the 750 mg group. Histopathology showed dose related effects of epithelial proliferation and inflammation of the forestomach already visible in animals treated at 150 mg.

There were no further major changes observed in other parameters (haematology, biochemistry, neurobehavioral functions, organ weights)

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.

 

Reproductive parameters were not affected. No effects were seen between the groups in duration of pregnancy, number of corpus luteum, number of implantation, pre-implantation loss, post-implantation loss. There was no significant difference between the control and treatment group in terms of copulation, fertility and gestation index.

Also the BW of the pups and external examination of pups (group) on post-natal day 1 and 4 show no effects.

Consequently the NOAEL for developmental effects was stated to be above the highest tested dose (750 mg/kg/day)

 

 

Tosyl chloride rapidly hydrolyses intoToluene-4-sulphonic acid and HCl, both strong acids. Systemic exposures will therefore actually be on Toluene-4-sulphonic acid. Consequently, cross-reading from Toluene-4-sulphonic acid for longer term repeated dose studies such as fertility and developmental toxicity can be applied. For Toluene-4-sulphonic acid, studies from the chemically related hydrotropes category are being recommended as read across for this endpoint. The following information is available for these substances:

No fertility studies are reported for the aromatic sulphonic acids. These substances are very corrosive as was demonstrated in skin and eye irritation studies, in the acute oral studies, and in the single repeated dose oral study. For animal welfare, long-term exposures to corrosive substances are not recommended. There are however studies for the chemically related hydrotrope substances that looked at reproductive organs and development of offspring. Hydrotropes are the salt form of the sulphonic acids and therefore are used as read-across for this endpoint. The 90-day oral rat and oral mouse studies and the 2-year chronic dermal rat and mouse studies with the closely related compound sodium xylene sulfonate (CAS No. 1300-72-7) included examination of sex organs of both sexes. No treatment related effects on reproductive organs were reported at doses roughly equivalent to those in the developmental toxicity study. The 1994 study with calcium xylene sulphonate (CAS No. 28088-63-3) did not follow a specific guideline but was fully documented and conducted in accordance with GLP requirements. No adverse effects were reported. The NOAEL for both maternal and foetal toxicity was the highest dose tested - 3000 mg/kg bw /day which is equivalent to 936 mg active ingredient per kilogram body weight per day. The conclusion of the study was no indications of developmental toxicity including teratogenesis.

 

The available data indicate a low concern for reproduction toxicity for Tosyl chloride. An OECD 422 screening study indicated no effects on any reproductive parameter up to 750 mg/kg, the highest dose tested. It should be remarked that this levels is 5 times the LOAEL level of 150 mg/kg based on local gastro-intestinal irritation with specific marked effects in the nonglandular stomach seen at this dose level. Also cross-reading from comparable structures do not indicate a concern for reproductive toxicity. When further considering the low likelihood of exposures followingthe use as a chemical intermediate in industrial or professional settings and as result of its physical-chemical properties, there is no need for further testing.

Short description of key information:

There were no reproductive effects seen in a Reproduction/Developmental Toxicity Screening Test according to OECD 422 at 750 mg/kg, the highest dose tested.

Justification for selection of Effect on fertility via oral route:

Only available data for reproduction on Tosyl chloride.

Justification for selection of Effect on fertility via inhalation route:

Exposures by inhalation are unlikely in view of the low vp of 0.13 Pa at 20°C, its low dustiness (the respirable fraction (≤ 4 µm) of the hygroscopic, crystalline solid is below 0,04%) and low likelihood of exposures by aerosols with uses limited to chemical intermediate in industrial settings. Furthermore Tosyl chloride readily hydrolyses to toluene sulfonic acid and HCL which are both corrosive acids. Inhalation of dust particles will therefore lead to respiratory irritation/corrosion rather than expected to result in systemic toxicity. Additionally it would technically be difficult to perform inhalation studies with Tosyl chloride dust in view of its hygroscopic properties.

Justification for selection of Effect on fertility via dermal route:

In view of irritation properties to skin and no effects observed via oral route which is considered a more efficient route of administration, there is no need to perform an additional study via dermal route to evaluate reproduction toxicity.

Effects on developmental toxicity

Description of key information

There were no developmental effects seen in a Reproduction/Developmental Toxicity Screening Test according to OECD 422 at 750 mg/kg, the highest dose tested. A developmental toxicity study on a structurally related substance also showed no developmental toxicity at the highest dose tested of 936 mg/kg.

Link to relevant study records

Reference

Reference 1

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

February 7-24, 1994

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Not a guideline study per se. GLP compliance. Full documentation

Qualifier:

no guideline followed

Principles of method if other than guideline:

Females were mated with males of the same strain and source. A block design was used to assign 30 mated females to controls and each of three dosing levels. Doses were selected based on a range finding study where the highest dose was 3000 mg/kg/day and no maternal or developmental toxicity was observed. The test material was prepared weekly. Analysis of dosing preparations stored for 10 days at room temperature verified the stability of the dosing preparation for at least a week. Dosing was done by intragastric intubation through needles at a volume of 10 m?/kg. Dosing was done on gestation days 6 through 15. A vehicle control was included in the study design. The concentration of the dosing preparation was confirmed analytically. Animals were observed twice daily for mortality and signs of overt toxicity. Clinical observations were made from gestation days - 20. Individual body weights were recorded on gestation days 0, 6, 9, 12, 16 and 20. Individual food consumption was recorded concurrent with the body weighing days. Food consumption was calculated. On gestation day 20 all surviving animals were euthanized and immediately examined by cesarean section. The uterus and overies were exposed and examined. The uterus was weighed. The location of viable and nonviable fetuses, early and late resorptions, and the number of total implantations and corpora lutea were recorded. The abdominal and thoracic cavities and organs were examined for grossly evident morphological changes. Uteri from nongravid females were placed in 10% ammonium sulfide solution for detection of implantations. Individual fetuses were weighed, sexed, and examined for external malformations and variations. Approximately one-half of the fetuses were placed in Bouin's solution for subsequent soft-tissue examination using hte Wilson razor-blade sectioning technique. The remainder of the fetuses were prepared for skeletal examination. All gross, visceral and skeletal alterations observed were classified as malformations or developmental variations. Treatment and control groups were subjected to appropriate statistical comparison.

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Species:

rat

Strain:

other: Charles River Crl:CD VAF/Plus

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Michigan
- Age at study initiation: 12.5 weeks
- Weight at study initiation: 243-312g
- Fasting period before study: no data
- Housing: individually in suspended stainless steel wire mesh cages
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 10 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 71-71 degrees F
- Humidity (%): 48-59%
- Air changes (per hr): controlled
- Photoperiod (hrs dark / hrs light): 12/12


IN-LIFE DATES: From: February 7 To: February 24

Route of administration:

oral: gavage

Vehicle:

not specified

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:

DIET PREPARATION
- Rate of preparation of diet (frequency): weekly
- Mixing appropriate amounts with (Type of food): Purina Certified Rodent Chow #5002
- Storage temperature of food: room temperature


VEHICLE
- Justification for use and choice of vehicle (if other than water): vehicle used but no data

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

no data

Details on mating procedure:

- Impregnation procedure: cohoused
- If cohoused:
- M/F ratio per cage: 1/1
- Length of cohabitation: until evidence of copulatory plug (7 days maximum)
- Verification of same strain and source of both sexes: yes
- Proof of pregnancy: vaginal plug
- Any other deviations from standard protocol: no data

Duration of treatment / exposure:

On gestation days 6-15

Frequency of treatment:

Daily

Duration of test:

Until gestation day 20

Remarks:

Doses / Concentrations:
150, 1500 and 3000 mg/kg/day
Basis:
analytical conc.

No. of animals per sex per dose:

30 females

Control animals:

yes, concurrent no treatment

Details on study design:

- Dose selection rationale: prior range finding study (study 715-001)
- Rationale for animal assignment (if not random): random block
- Other:

Maternal examinations:

Animals were observed twice daily for mortality and signs of overt toxicity. Clinical observations were made from gestation days - 20. Individual body weights were recorded on gestation days 0, 6, 9, 12, 16 and 20. Individual food consumption was recorded concurrent with the body weighing days. Food consumption was calculated. On gestation day 20 all surviving animals were euthanized and immediately examined by cesarean section.

Ovaries and uterine content:

The uterus and overies were exposed and examined. The uterus was weighed. The location of viable and nonviable fetuses, early and late resorptions, and the number of total implantations and corpora lutea were recorded. The abdominal and thoracic cavities and organs were examined for grossly evident morphological changes. Uteri from nongravid females were placed in 10% ammonium sulfide solution for detection of implantations.

Fetal examinations:

Individual fetuses were weighed, sexed, and examined for external malformations and variations. Approximately one-half of the fetuses were placed in Bouin's solution for subsequent soft-tissue examination using hte Wilson razor-blade sectioning technique. The remainder of the fetuses were prepared for skeletal examination. All gross, visceral and skeletal alterations observed were classified as malformations or developmental variations.

Statistics:

Treatment and control groups were subjected to appropriate statistical comparison. Dunnett t-test for body wight, food consumption, numbers of corpora lutea, total implantations, live fetuses and mean fetal body weights. Chi-square or Fishers test for male to female sex ratios and proportion of litters with malformations and developmental variations. Kruskal-Wallis test for the proportion of resorbed and dead fetuses and postimplantation losses

Indices:

see "Statistics" above

Details on maternal toxic effects:

Maternal toxic effects:no effects

Details on maternal toxic effects:
One death occurred at the 1500 mg/kg/day dose but it was considered a gavage injury. No clinical observations or necropsy findings. No effects on body weight or body weight gain. There was a significant increase in food consumption for the 3000 mg/kg/day during gestation interval 12-16 but this was considered normal biological variation and not a direct effect of the test substance.

Key result

Dose descriptor:

NOAEL

Effect level:

> 3 000 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

other: other:

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
No indications of developmental toxicity including teratogenesis. All indices were comaparable to the corresponding controls.

Key result

Dose descriptor:

NOAEL

Effect level:

> 3 000 mg/kg bw/day

Based on:

test mat.

Sex:

not specified

Basis for effect level:

other:

Remarks on result:

other: No indications of developmental toxicity including teratogenesis

Abnormalities:

not specified

Developmental effects observed:

not specified

Conclusions:

The test substance did not induce developmental or teratogenic effects at doses up to 3000 mg/kg/day.

Executive summary:

Thirty (30) female rats received 0, 150, 1500 or 3000 mg test substance per kilogram body weight by oral gavage on days 6 to 15 of gestation. Clinical symptoms were noted daily through day 20. Body weight and food consumption were recorded every three days through day 20. All females were macroscopically examined on day 20. The uteri were removed, weighed and examined for number of corpora lutea, implant sites, and number and location of fetuses and resorptions. Fetuses were inspected on total number, sex, weight and external, visceral (one-half) and skeletal (one-half) defects. One death occurred at the 1500 mg/kg/day dose but it was considered a gavage injury. There were no abnormal clinical observations or necropsy findings. There were no effects on body weight or body weight gain. There was a significant increase in food consumption for the 3000 mg/kg/day dose during gestation interval (day) 12-16 but this was considered normal biological variation and not a direct effect of the test substance. All indices were comparable to the corresponding controls. The NOAEL based on active ingredient of the test substance is 936 mg/kg bw/day.

Reference 2

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

supporting 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.

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

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

Route of administration:

oral: gavage

Vehicle:

not specified

Analytical verification of doses or concentrations:

not specified

Details on mating procedure:

Each male and female rat was selected from the same test group in order to copulate. It spent 14 days in terms of mating. The day after the copulating, mating would be determined through the observation of sperm in a vaginal rinse.

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

Duration of test:

34-51, covering the full duration of pregnancy

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 no treatment

Details on study design:

According to the preliminary test (Report No. P104), dose level was set to 0, 150, 350 and 750 mg/kg/day.

Maternal examinations:

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

Ovaries and uterine content:

The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: No
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes

Fetal examinations:

Body weight of pups
Examination of the external surface of pups: It was observed on the day of delivery and on the day 4 of lactation to determine abnormalities.

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.

Indices:

Reproductive indices:
- Copulation index
- number of implantations
- number of corpus luteum
- Number of females pregnant
Offspring viability indices:
- Number of life pups
- post natal loss
- pre and post-implantation loss

Details on maternal toxic effects:

Maternal toxic effects:yes. Remark: at 150 mg/kg (lowest dose)

Details on maternal toxic effects:
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.

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.

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.

GROSS PATHOLOGY
No information provided.

HISTOPATHOLOGY:
dose related effects of epithelial proliferation and inflammation of the forestomach already visible in animals treated at 150 mg.

Dose descriptor:

NOAEL

Effect level:

750 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

other: developmental toxicity

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
Number of pups born, viability and sex ratio: No significant difference was observed between the treatment groups and the control group at the time of the delivery and on the day 4 of the lactation.
External examination of pups (group) showed no developmental abnormalities.

Abnormalities:

not specified

Developmental effects observed:

not specified

Body weights of pups (group)

Group	Sex	Pup BW at birth		At lactation day 4
		Male	Female	Male	Female
0	Mean	6.25	5.75	8.57	8.03
	SD	0.6	0.66	1.26	1.18
	N	67	69	66	69
150	Mean	5.99	5.62	8.56	7.98
	SD	0.56	0.46	1.34	1.27
	N	62	70	63	66
350	Mean	6.37	5.73	9.71	8.59
	SD	0.9	0.8	2.17	1.57
	N	59	48	55	44
750	Mean	5.98	5.59	8.7	8.21
	SD	0.6	0.51	0.87	0.74
	N	48	51	46	49

 

External examination of pups (group)

Group	No. of animals	At birth			No. of animals	Lactation day 4
		No.of pups examined	No.of pups	Obser-vations		No.of pups examined	No.of pups	Obser-vations
0	11	143	140	NGF	11	136	134	NGF
			2	BT			2	BT
			1	Runt
150	11	143	144	NGF	11	129	129	NGF
350	10	116	116	NGF	9	100	100	NGF
750	9	105	105	NGF	8	95	95	NGF

NGF: No Gross findings

BT: Blunt-tipped Tail

Conclusions:

There were no reproductive or developmental effects seen at 750 mg/kg, the highest dose tested

Executive summary:

Tosyl chloride has been evaluated in a Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test according to OECD 422 and in compliance to GLP.

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.

Results:

Parental toxicity: No mortality males; in females 350 mg/kg: 2/12 and at 750 mg/kg: 4/18.

Some clinical signs were observed at the dose level of 150 mg/kg/day in both male and females such as intermittent (blood-like) salivation and staining around mouth. Effects were more severe at 350 mg/kg and at 750 mg/kg reported clinical signs were 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. A decreased BW gain was observed in the 750 mg group resulting to a lower BW that was maximal 8% lower in males and 5.5% in females compared to controls. Food consumption was a bit lower during the first week in the 750 mg group. Histopathology showed dose related effects of epithelial proliferation and inflammation of the forestomach already visible in animals treated at 150 mg.

There were no further major changes observed in other parameters (haematology, biochemistry, neurobehavioral functions, organ weights)

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.

 

Reproductive parameters were not affected.No effects were seen between the groups in duration of pregnancy, number of corpus luteum, number of implantation, pre-implantation loss, post-implantation loss. There was no significant difference between the control and treatment group in terms of copulation, fertility and gestation index.

Also the BW of the pups and external examination of pups (group) on post-natal day 1 and 4 show no effects.

Consequently the NOAEL for developmental effects was stated to be above the highest tested dose (750 mg/kg/day)

Reference 3

Endpoint:

developmental toxicity

Type of information:

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

Adequacy of study:

key study

Justification for type of information:

Also attached :

Category of Aromatic Sulphonic Acids (ASA)

The aromatic sulphonic acids category consists of the following five (5) substances:

• Benzene sulphonic acid (CAS No. 98-11-3)
• 4-hydroxybenzene sulphonic acid (CAS No. 98-67-9)
• Toluene-4-sulphonic acid (CAS No. 104-15-4)
• Xylene sulphonic acid (CAS No. 25321-41-9)
• p-cumene sulphonic acid (CAS No. 16066-35-6)


Sulphonic acids are a class of organic acids with the general formula R-S(=O)2-OH, where R is usually a hydrocarbon (aromatic) side chain. The sulphonic acids are strong acids and in watery environments they are almost completely ionized even at low pH.

The five aromatic sulphonic acids in this category are chemically related and quite similar. Benzenesulphonic acid is an organic compound with the formula C6H5SO3H. It is the simplest aromatic sulphonic acid. Benzene with a hydroxyl group is hydroxybenzene (or phenol). Benzene with a methyl group is toluene. Benzene with two methyl groups is xylene. And benzene with an isopropyl group is cumene.
Acidity of the sulphonic acid group is influenced by two factors:

1. The substitution group exerts an electron donating effect and so makes the negative charge on the resulting sulphonate ion, after deprotonation, more or less stable.
2. The resonance effect stabilizes the negative charge on the sulphonate ion by dividing the charge on the oxygen atoms.

As a result, the acidity of the sulphonic acid group is not expected to change significantly among the five aromatic sulphonic acids. The measured pKa values confirm this expectation: 2.68 for benzene sulphonic acid, 2.50 for hydroxybenzene sulphonic acid, 2.96 for toluene-4-sulphonic acid, 2.88 for xylene sulphonic acid and 3.17 for cumene sulphonic acid. Thus the reactivity of the sulphonic acids are very similar and they can each be used as a surrogate for the others.
A full set of 2010 guideline physical-chemical studies demonstrates the similar chemical and physical properties and behavior of the 5 sulphonic acids in the category.
The sulphonic acid moiety is the primary driver for mammalian toxicity and any difference between the benzene, xylene, cumene, and toluene moieties would be insignificant given the relatively high level of corrosivity of all five substances in the category.
As noted, the aromatic sulphonic acids are almost completely ionized in watery environments.
These acids are precursors to hydrotropes (or “sulphonates”) which include ammonium, calcium, potassium and sodium salts. Because of the chemical similarities and because much of the production of the aromatic sulphonic acids goes to manufacturing the salts, the extensive database for the hydrotropes is also used as a source of read-across for some of the endpoints in this aromatic sulphonic acid dossier.

Read across from Hydrotropes to Aromatic Sulphonic Acids
The studies with the salts (hydrotropes) provide valid read-across for the acids. The specific cation is not expected to have an appreciable effect on fate, ecotoxicity or mammalian toxicity and therefore the dataset for the entire hydrotropes category can be applied broadly.
The aromatic sulphonic acids are almost completely ionized in watery environments even at low pH. The salts of these acids are the hydrotropes (or “sulphonates”) which include ammonium, calcium, potassium and sodium cations. In principle the salts get dissociated when in contact with water, so forming back to the acids. Because of their close chemical similarities and because much of the production of the aromatic sulphonic acids goes to manufacturing the salts, the extensive dataset for the hydrotropes can also be used as a source of read-across for endpoints in an aromatic sulphonic acid dossier. This is particularly relevant for studies that are conducted in water (e.g., ecotoxicity and biodegradation) as well as for mammalian toxicity studies where the relatively high acidity of the acid form has an immediate and harsh local effect, whereas the salt form provides an indication of potential systemic toxicity beyond the site of application or initial contact

Reason / purpose for cross-reference:

read-across source

Details on maternal toxic effects:

Maternal toxic effects:no effects

Details on maternal toxic effects:
One death occurred at the 1500 mg/kg/day dose but it was considered a gavage injury. No clinical observations or necropsy findings. No effects on body weight or body weight gain. There was a significant increase in food consumption for the 3000 mg/kg/day during gestation interval 12-16 but this was considered normal biological variation and not a direct effect of the test substance.

Key result

Dose descriptor:

NOAEL

Effect level:

> 3 000 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

other: other:

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
No indications of developmental toxicity including teratogenesis. All indices were comaparable to the corresponding controls.

Dose descriptor:

NOAEL

Effect level:

> 3 000 mg/kg bw/day

Based on:

test mat.

Sex:

not specified

Basis for effect level:

other:

Remarks on result:

other: No indications of developmental toxicity including teratogenesis

Abnormalities:

not specified

Developmental effects observed:

not specified

Conclusions:

The test substance Calcium xylene sulponate did not induce developmental or teratogenic effects at doses up to 3000 mg/kg/day. This conclusion is read acrossed to p-Toluene sulfonyl chloride.

Executive summary:

Thirty (30) female rats received 0, 150, 1500 or 3000 mg test substance per kilogram body weight by oral gavage on days 6 to 15 of gestation. Clinical symptoms were noted daily through day 20. Body weight and food consumption were recorded every three days through day 20. All females were macroscopically examined on day 20. The uteri were removed, weighed and examined for number of corpora lutea, implant sites, and number and location of fetuses and resorptions. Fetuses were inspected on total number, sex, weight and external, visceral (one-half) and skeletal (one-half) defects. One death occurred at the 1500 mg/kg/day dose but it was considered a gavage injury. There were no abnormal clinical observations or necropsy findings. There were no effects on body weight or body weight gain. There was a significant increase in food consumption for the 3000 mg/kg/day dose during gestation interval (day) 12-16 but this was considered normal biological variation and not a direct effect of the test substance. All indices were comparable to the corresponding controls. The NOAEL based on active ingredient of the test substance is 936 mg/kg bw/day.

Based on the results of this study, the following NOAEL of Calcium xylene sulphonate was established, and read acrossed to p-Toluene Sulfonyl Chloride:

Developmental NOAEL 936 mg/kg bw/day.

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

936 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

Most relevant GLP compliant study available, evaluating developmental toxicity, based on cross-reading.

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no study available

Additional information

Tosyl chloride has been evaluated in a Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test according to OECD 422 and in compliance to GLP. (See under reproduction toxicity)

Reproductive parameters were not affected. No effects were seen between the groups in duration of pregnancy, number of corpus luteum, number of implantation, pre-implantation loss, post-implantation loss. There was no significant difference between the control and treatment group in terms of copulation, fertility and gestation index.Also the BW of the pups and external examination of pups (group) on post-natal day 1 and 4 show no effects.

Consequently the NOAEL for developmental effects was stated to be above the highest tested dose (750 mg/kg/day)

 

Tosyl chloride rapidly hydrolyses intoToluene-4-sulphonic acid and HCl, both strong acids. Systemic exposures will therefore actually be on Toluene-4-sulphonic acid. Consequently, cross-reading from Toluene-4-sulphonic acid for longer term repeated dose studies such as fertility and developmental toxicity can be applied. For Toluene-4-sulphonic acid, studies from the chemically related hydrotropes category are being recommended as read across for this endpoint. The following information is available for these substances:

 

A developmental toxicity study comparable to OECD 414 was performed with on the structurally related substance Calcium xylenesulphonate.

In the developmental toxicity study, thirty (30) female rats received 0, 150, 1500 or 3000 mg test substance per kilogram body weight by oral gavage on days 6 to 15 of gestation. Clinical symptoms were noted daily through day 20. Body weight and food consumption were recorded every three days through day 20. All females were macroscopically examined on day 20. The uteri were removed, weighed and examined for number of corpora lutea, implant sites, and number and location of foetuses and resorptions. Foetuses were inspected on total number, sex, weight and external, visceral (one-half) and skeletal (one-half) defects. One death occurred at the 1500 mg/kg/day dose but it was considered a gavage injury. There were no abnormal clinical observations or necropsy findings. There were no effects on body weight or body weight gain. There was a significant increase in food consumption for the 3000 mg/kg/day dose during gestation interval (day) 12-16 but this was considered normal biological variation and not a direct effect of the test substance. All indices were comparable to the corresponding controls. The NOAEL based on active ingredient of the test substance is 936 mg/kg bw/day.(On molar basis equivalent to 870 mg Tosyl chloride/kg bw.

 

The available data indicate a low concern for developmental toxicity for Tosyl chloride. An OECD 422 screening study indicated no effects on any reproductive parameter up to 750 mg/kg, the highest dose tested. It should be remarked that this levels is 5 times the LOAEL level of 150 mg/kg based on local gastro-intestinal irritation with specific marked effects in the nonglandular stomach seen at this dose level. Also cross-reading to developmental toxicity study comparable to OECD 414 on a comparable structure does not indicate a concern for developmental toxicity. When further considering the low likelihood of exposures followingthe use as a chemical intermediate in industrial or professional settings and as result of its physical-chemical properties, there is no need for further testing.

Justification for selection of Effect on developmental toxicity: via oral route:

Most relevant available study evaluating developmental toxicity, based on cross-reading.

Justification for selection of Effect on developmental toxicity: via inhalation route:

Not preferred route for testing (See fertility)

Justification for selection of Effect on developmental toxicity: via dermal route:

Not preferred route for testing (See fertility)

Justification for classification or non-classification

Available data do not indicate a concern for reproduction or developmental toxicity.