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Description of key information

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1999
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was performed in accordance with OECD guideline 401 and GLP standard.
Reason / purpose:
reference to same study
Qualifier:
according to
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Deviations:
not specified
Principles of method if other than guideline:
NA
GLP compliance:
yes
Test type:
standard acute method
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: no data
- Age at study initiation: 5 weeks
- Weight at study initiation: 157-197g for male, 132-151g for female
- Fasting period before study: Rats were fasted for approximately 16 hours prior to dosing the test substance, but drinking water was freely ingested in the cage.
- Housing: no data
- Diet (e.g. ad libitum): no data
- Water (e.g. ad libitum): ad libitum
- Acclimation period: no data

ENVIRONMENTAL CONDITIONS
- Temperature (°C): no data
- Humidity (%): no data
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): no data

IN-LIFE DATES: no data
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle:
- Amount of vehicle (if gavage):
- Justification for choice of vehicle:
- Lot/batch no. (if required):
- Purity:

MAXIMUM DOSE VOLUME APPLIED:

DOSAGE PREPARATION (if unusual):

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose:
Doses:
0, 1000, 2000 and 3000 mg/kg
No. of animals per sex per dose:
5 males and 5 females per dose group
Control animals:
yes
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: The animals were observed for clinical signs of toxicity just before the administration at 1, 2, 3, 4, 5, and 6 hrs after dosing on day 1 and once a day until day 14. Individual body weights of animals were measured just before the administration, on day 3, 7 and the day of necropsy.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,necropsy
Statistics:
The body weight changes were analyzed using the Student’s t-test.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
>= 1 000 - <= 2 000 mg/kg bw
Mortality:
During the observation period, all males and four females died at both 2,000 and 3,000 mg/kg bw.
Clinical signs:
In case of dead rats, the paralysis of forelimb and hindlimb, cloudy eyeball, and lacrimation on day 2 after the administration were observed in those groups. The slight paralysis of hindlimb was observed in 2 males and 2 females at 1,000 mg/kg bw and 1 female rat at 2,000 mg/kg bw on day 2. In female rats, however, these signs were recovered on day 9 and 14, respectively. The other animals were not observed any sign related with the substance treatment.
Body weight:
At 2,000 and 3,000 mg/kg bw dose group, the body weights for male and female rats were significantly lower than those of the control groups on day 3.
Gross pathology:
Dead animals of the 2,000 and 3,000 mg/kg groups showed dark-red discoloration of the brain and lung, and the atrophic thymus and spleen. In scheduled necropsy, however, no macroscopic abnormalities were seen in the surviving animals.
Other findings:
None

Table1. Mortality and clinical signsof the acute oral toxicity

Sex

Group

No. of animals

Findings

Days after treatment

1

2

3

4

5

6

7

8

9

10

11

12

13

14

Male

C

5

N

5

5

5

5

5

5

5

5

5

5

5

5

5

5

T1

5

N

5

5

5

4

3

4

4

3

3

3

3

3

3

3

P*

 

 

 

1

2

1

1

2

2

2

2

2

2

2

T2

5

N

5

 

 

 

 

 

 

 

 

 

 

 

 

 

P

 

5

5

5

5

 

 

 

 

 

 

 

 

 

C

 

5

4

5

5

 

 

 

 

 

 

 

 

 

L

 

 

 

5

5

 

 

 

 

 

 

 

 

 

D

 

 

 

 

 

5

 

 

 

 

 

 

 

 

T3

5

N

5

 

1

1

1

 

 

 

 

 

 

 

 

 

P

 

5

4

1

1

1

 

 

 

 

 

 

 

 

C

 

 

2

1

1

1

1

 

 

 

 

 

 

 

H

 

 

 

 

 

1

1

 

 

 

 

 

 

 

L

 

 

1

1

1

1

 

 

 

 

 

 

 

 

D

 

 

 

3

 

1

 

1

 

 

 

 

 

 

Female

C

5

N

5

5

5

5

5

5

5

5

5

5

5

5

5

5

T1

5

N

5

4

4

3

3

3

3

4

5

5

5

5

5

5

P*

 

1

1

2

2

2

2

1

 

 

 

 

 

 

T2

5

N

5

 

 

 

 

 

 

 

 

 

 

 

 

1

P

 

5

4

2

1

 

 

 

 

 

 

 

 

 

C

 

 

1

 

 

 

 

 

 

 

 

 

 

 

P*

 

 

 

1

2

1

1

1

1

1

1

1

1

 

D

 

 

1

1

 

2

 

 

 

 

 

 

 

 

T3

5

N

5

2

2

1

1

1

1

1

1

1

1

1

1

1

P

 

3

3

4

3

1

 

 

 

 

 

 

 

 

D

 

 

 

 

1

2

1

 

 

 

 

 

 

 

Table 2. Body weights of male and female rats

Group

 

Mean body weight (g) of males

Mean body weight (g) of females

Day 0

Day 3

Day 7

Day 14

Day 0

Day 3

Day 7

Day 14

C

Mean

153.3

194.7

232.1

296.6

125.1

153.2

172.8

195.7

 

S.D.

11.1

12.0

15.1

20.7

4.2

6.9

8.6

13.3

 

N

   5

 5

   5

   5

   5

   5

  5

  5

T1

Mean

160.9

172.2

201.8

274.6

127.7

142.0

165.1

196.5

 

S.D.

7.1

23.9

41.8

39.2

5.0

10.4

7.1

9.8

 

N

  5

 5

   5

   5

   5

   5

  5

   5

T2

Mean

163.7

141.1**

0.0

      0.0

123.2

109.6**

120.4

162.4

 

S.D.

8.0

10.9

0.0

      0.0

5.7

3.8

      0.0

      0.0

 

N

5

5

  0

  0

5

5

1

1

T3

Mean

160.2

134.4**

108.6

0.0

128.0

123.8*

186.0

215.0

 

S.D.

9.3

7.1

0.0

0.0

5.9

19.4

0.0

0.0

 

N

5

5

1

   0

5

5

1

1

Conclusions:
The acute oral toxicity of OBSH was assessed in rats following a single administration of OBSH. The study was performed in compliance with OECD guideline 401. The acute oral median lethal dose (LD50) of OBSH was estimated to be between 1000 and 2000 mg/kg bw under the test conditions.
According to CLP criteria OBSH should be classified as Acute tox 4; H302.
Executive summary:

This study was conducted to assess the acute toxicity of OBSH following a single oral administration of 0, 1000, 2000 and 3000 mg/kg bw. This study is performed in accordance with the testing guideline OECD (401).

Groups of five males and five female fasted rats were given OBSH as a single dose on Day 1 by oral gavage at a dose level of 0, 1000, 2000 and 3000 mg/kg. OBSH was administrated in corn oil.

Following a single administration of OBSH to rats, the acute median lethal oral dose for male and female rats was estimated to be between 1000 and 2000 mg/kg bw under the test conditions.

According to CLP criteria OBSH should be classified as Acute tox 4; H302.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
1 500 mg/kg bw
Quality of whole database:
The study was performed in accordance with OECD guideline 401 and GLP standard - Klimisch score 1.

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
March 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 436 (Acute Inhalation Toxicity: Acute Toxic Class Method)
Deviations:
no
Principles of method if other than guideline:
NA
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
no
Specific details on test material used for the study:
Identity: 4,4’-Oxybis(benzenesulfonyl hydrazide) (OBSH)
CAS No. : 80-51-3
EC No.: 201-286-1
Test facility Code No.: K-5450
Batch/Lot No.: 201512001
Appearance White fine powder
Purity: 99.5 - 100%
Storage conditions: Room temperature, Protected from light, Protected from moisture
Expiry date: 2016.12
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
Species Rat (Specific Pathogen Free)
Strain/Substrain Sprague-Dawley/Crl:CD (SD)
Number at receipt 20 (Male 10, Female 10)
Number on study start 12 (Male 6, Female 6)
Number on study end 12 (Male 6, Female 6)
Age range at receipt Approximately 8 weeks
Age range on study start Approximately 9 weeks (1st step) Approximately 10 weeks (2nd step) '
Body weight range Male: 312.4-337.2 g at the start of exposure (1st step) Male: 341.6-359.4 g at the start of exposure (2nd step) Female: 196.0-213.5 g at the start of exposure (1st step) Female: 204.1-217.6 g at the start of exposure (2nd step)

Supplier Orient Bio Inc. 322, Galmachi-ro, Jungwon-gu, Seongnam-si, Gyeonggi-do 13201, Republic of Korea Method of identification Color marking, Tail tattoo, Cage card
Acclimation period 5 days Pre-treatment period 2 days (1st step) 6 days (2nd step)
Replacement There was no replacement of the animals.

During pre-treatment period, 1 time of holder adaptation training was performed according to KIT SOP in order to decrease a stress that could be resulted from nose-only inhalation exposure

Housing and Animal Care:

Animal and exposure room No.
Animal room No.: F103, Exposure room No.: F102 (Inhalation toxicology building)

Housing
Animals were housed in groups of up to 3 animals in suspended, stainless-steel cage (255W×465L×200H mm) for the acclimation period, pre-treatment and exposure period.

Environment
The animal room environment was automatically controlled according to SOPs (target range: temperature 22 ± 3°C, relative humidity 30-70%, approximately 12 hours light cycle with 150-300 Lux, and ventilation 10-20 times/hour). Temperature and relative humidity were monitored continuously. Animal room and cage cleaning was performed according to KIT SOPs.

Identification for cage and animal room and exposure room
Cage card were attached to the cages. The animal room use record was displayed in the animal room and the exposure room use record use record was also displayed in the exposure room.

Food and Water
A standard rat and mouse pellet diet (Lab Diet® #5053, PMI Nutrition International, USA; irradiated by gamma-ray) was provided to the animals ad libitum. Microbial monitoring for diet was performed at test facility and a certificate of analysis for the diet was provided by the supplier. The animals have had ad libitum access to filtered, ultraviolet light-irradiated municipal tap water at all times. The drinking water was analyzed every 6 months for specified contaminants in the Gwangju Health & Environment Research Institute (149, Hwajeong-ro, Seo-gu, Gwangju, 61986, Republic of Korea). There were no known contaminants in the food or water at levels that would be expected to interfere with the results of the study and the data were maintained in the raw data.
Route of administration:
inhalation: dust
Type of inhalation exposure:
nose only
Vehicle:
not specified
Remark on MMAD/GSD:
Mass median aerodynamic diameters (MMADs) were 2.22 and 2.26 μm and geometric standard deviations (GSDs) were 2.02 and 1.92 for 1st step exposure, respectively.

Mass median aerodynamic diameters (MMADs) were 2.47 and 2.48 μm and geometric standard deviations (GSDs) were 2.00 and 2.03 for 2nd step exposure, respectively.
Details on inhalation exposure:
Exposure method of test item:
Test item was generated using dust generator [DF-3 Special, Sibata] and then supplied to the nose-only exposure inhalation experiment device [SIS-30BN, Sibata].

Chamber and conditions inside chamber:
Nose-only exposure inhalation experiment device [SIS-30BN, Sibata] Temperature was targeted to be maintained at 22 ± 3°C and relative humidity was targeted to be maintained in the range of 50 ± 20%. As well, oxygen concentration was targeted to be maintained above 19% and carbon dioxide concentration was targeted to be maintained below 1%.

Measurement and Monitoring of Exposure:
Nominal concentration: Total amount of test items used for generation of test item / Total volume of air supplied into chamber
Actual concentration: Exposure concentration of test item was measured 3 times by gravimetric analysis using microglass fiber filter during exposure, and then actual concentration (mass concentration) of exposed test item was assessed.

Environmental conditions of inhalation chamber:
Temperature, relative humidity, chamber airflow, chamber pressure, oxygen concentration and carbon dioxide concentration were monitored and recorded at least hourly during exposure.
Oxygen concentration was measured by multi-gas monitor [M40, ISC] and carbon dioxide concentration was measured by carbon dioxide meter [PGM-6208, RAE Systems].
Particle size distribution:
Particle size of the generated and exposed test item was measured 2 times during exposure by cascade impactor [Mini MOUDI 135-6S, MSP Corporation].
Mass median aerodynamic diameter (MMAD) with geometric standard deviation (GSD) was assessed based on measured values using Microsoft Excel® software (Microsoft Corporation, USA).

Method Verification (Inhalation Exposure Validation):
The chamber homogeneity on generated and exposed test item was performed during acclimation period.
Analytical verification of test atmosphere concentrations:
yes
Duration of exposure:
4 h
Concentrations:
The rats were assigned to two groups (3 animals/sex/group) and were exposed nose-only to target concentrations of 1.00 mg/L (1st step exposure) or 5.00 mg/L (2nd step exposure) of 4,4’-Oxybis(benzenesulfonyl hydrazide) (OBSH) for 4 hours.

Mean exposure concentration (actual concentration) of test item of 1st step and 2nd step were 1.06 ± 0.07 mg/L and 4.78 ± 0.30 mg/L for 4 hours, respectively
No. of animals per sex per dose:
3 animals per sex per dose
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: At receipt, at randomization, on day 1, 2, 5, 8 and 14 and at necropsy.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight and macrosopic findings
Key result
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 5 mg/L air (analytical)
Based on:
test mat.
Exp. duration:
4 h
Remarks on result:
not determinable due to absence of adverse toxic effects
Mortality:
There were no unscheduled deaths in both sexes of 1st or 2nd step exposure group during the study period.
Clinical signs:
other: There were no test item-related clinical signs in both sexes of 1st or 2nd step exposure group during the study period
Body weight:
After acute inhalation exposure, body weights in both sexes of 1st and 2nd step exposure groups were transiently decreased on Day 2. Body weight gains in both sexes of 1st and 2nd step exposure groups were also transiently delayed on Day 2. But afterward, these were recovered.
Gross pathology:
There were no test item-related gross changes in both sexes of 1st or 2nd step exposure group
Other findings:
NA
Interpretation of results:
GHS criteria not met
Conclusions:
The acute inhalation toxicity of 4,4’Oxybis(benzenesulfonyl hydrazide) (OBSH) was evaluated in accordance to OECD 436 in both sexes of Sprague-Dawley rats. The rats were assigned to two groups (3 animals/sex/group) and were exposed nose-only to target concentrations of 1.00 mg/L (1st step exposure) or 5.00 mg/L (2nd step exposure). No effects were seen, thus the LC50 was > 5.00 mg/L (actual concentration 4.78 mg/L) and GHS criteria for classification was not meet.
Executive summary:

The acute inhalation toxicity of 4,4’Oxybis(benzenesulfonyl hydrazide) (OBSH) was evaluated in accordance to OECD 436 in both sexes of Sprague-Dawley rats. The rats were assigned to two groups (3 animals/sex/group) and were exposed nose-only to target concentrations of 1.00 mg/L (1st step exposure) or 5.00 mg/L (2nd step exposure) for 4 hours.

The mean exposure concentration (actual concentration) of OBSH for the 1st step and 2nd step were 1.06 ± 0.07 mg/L and 4.78 ± 0.30 mg/L for 4 hours, respectively. Mass median aerodynamic diameters (MMADs) were 2.22 and 2.26 μm and geometric standard deviations (GSDs) were 2.02 and 1.92 for 1st step exposure, respectively. Mass median aerodynamic diameters (MMADs) were 2.47 and 2.48 μm and geometric standard deviations (GSDs) were 2.00 and 2.03 for 2nd step exposure, respectively.  

No unscheduled death were seen as well as no clinical signs or gross changes. Transient body weight decreases were observed in males and females of 1st and 2nd exposure groups at Day 2 but it was recovered afterward.

In conclusion, no effects were seen, thus the LC50 was > 5.00 mg/L (actual concentration 4.78 mg/L) and GHS criteria for classification was not meet in this study.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LC50
Quality of whole database:
A recently performed OECD 436 study performed in accordance to GLP standards - Klimisch score 1.

Acute toxicity: via dermal route

Endpoint conclusion
Endpoint conclusion:
no study available
Quality of whole database:
No reliable dermal acute toxicity study is available. A dermal toxicity study (reliability score 4) is available using 200 mg/kg of OBSH. No effeccts were seen after 24 hours. The original reference is not available. This study was not conducted according to GLP standard.

Additional information

The acute oral toxicity of OBSH was assessed in rats following a single administration of OBSH. Two studies were performed in compliance with OECD guideline 401.

In one study, groups of five males and five female fasted rats were given OBSH as a single dose by oral gavage at a dose level of 0, 1000, 1500 and 2000 mg/kg. OBSH was administered in corn oil. LD50 was just above 2000 mg/g bw. In another study, groups of five males and five female fasted rats were given OBSH as a single dose on Day 1 by oral gavage at a dose level of 0, 1000, 2000 and 3000 mg/kg. OBSH was administered in corn oil. Based on the results from these studies, the acute oral median lethal dose (LD50) of OBSH was estimated to be between 1000 and 2000 mg/kg bw under the test conditions.

The acute inhalation toxicity of OBSH was evaluated in accordance to OECD 436 in both sexes of Sprague-Dawley rats. The rats were assigned to two groups (3 animals/sex/group) and were exposed nose-only to target concentrations of 1.00 mg/L (1st step exposure) or 5.00 mg/L (2nd step exposure). No effects were seen, thus the LC50 was > 5.00 mg/L (actual concentration 4.78 mg/L) and GHS criteria for classification was not meet.

No reliable dermal acute toxicity study is available.


Justification for selection of acute toxicity – oral endpoint
The study is a key study

Justification for selection of acute toxicity – inhalation endpoint

The study is a key study


Justification for selection of acute toxicity – dermal endpoint
A dermal toxicity study (reliability score 4) is available using 200 mg/kg of OBSH. No effeccts were seen after 24 hours. The original reference is not available. This study was not conducted according to GLP standard.

Justification for classification or non-classification

According to CLP criteria OBSH should due to the oral LD50 value in the range of 1000-2000 mg/kg bw be classified as Acute tox 4; H302.