Registration Dossier

Diss Factsheets

Administrative data

Description of key information

Oral, inhalation and dermal acute toxicity are all considered.

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:
supporting study
Study period:
August 7 1978 to September 28th 1978
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Non GLP, not carried out according to recognised guideline, although results documented.
Reason / purpose for cross-reference:
reference to same study
Qualifier:
no guideline followed
Principles of method if other than guideline:
Acute oral toxicity in rats was determined according to the procedure suggested by Hagan. (Hagan E.C., (1959) Acute Toxicity; Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics, pp. 17 - 25). The rats were dosed individually by gavage, at graded dose levels (following range finding) after which they were returned to quarters where food and water were available ad libitum. Animals were observed for signs of pharmacologic activity and drug toxicity at 1, 3, 6 and 24 hours postdosage. Observations were made daily thereafter to a total of fourteen days. Animals sacrificed at the end of the 14-day observation period were subjected to complete gross necropsy.

LD50 was calculated, (including the 95% confidence limits where possible) using the method of Litchfield and Wilcoxin except when the highest maximum dose mechanically possible (40 ml/kg) is used.
GLP compliance:
no
Test type:
standard acute method
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
Wistar-derived albino rats, as indicated in the summary were assigned to groups, and maintained under standard laboratory conditions for a minimum of seven days, and fasted overnight prior to administration of the test material. The rats were dosed individually by gavage, at graded dose levels (following range finding) after which they were returned to quarters where food and water were available ad libitum. Animals are received from Summit View Farm, Belvidere, New Jersey and are conditioned prior to use.
Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
Albino rats in groups of ten (5M:5F), 150 - 300 g, single dosed orally, (40 ml/kg top dose mechanically feasible) observed fourteen days. Material
used as received. (Specific Gravity: 1.20)
Doses:
10.00, 20.00, 22.40, 25.20, 31.75, 40.00 g/kg body weight
No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
Animals were observed for signs of pharmacologic activity and drug toxicity at 1, 3, 6 and 24 hours postdosage. Observations were made daily thereafter to a total of fourteen days. Animals sacrificed at the end of the 14-day observation period were subjected to complete gross necropsy.
Statistics:
LD50 was calculated, (including the 95% confidence limits) where possible, using the method of Litchfield and Wilcoxin except when the highest maximum dose mechanically possible (40 ml/kg) is used. (Litchfield, J.T. and Wilcoxin, F. (1949) J. Pharmacol. Exptl. Therap. pp. 96, 99.)
Preliminary study:
Not applicable.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
15 750 mg/kg bw
Mortality:
See Table 1 below
Clinical signs:
Toxic effects noted were reddening of the pyloric and intestinal mucosa and matted, unkempt hair.
Body weight:
Not specified.
Gross pathology:
Intestines were very slightly reddened.

Table 1

Dose level

(g/kg)

Sex

Dead/dosed

%

10.00

5M:5F

1/5:1/5

20

20.00

5M:5F

5/5:1/5

60

22.40

5M:5F

5/5:5/5

100

25.20

5M:5F

4/5:5/5

90

31.75

5M:5F

5/5:5/5

100

40.00

5M:5F

5/5:4/5

90

Interpretation of results:
practically nontoxic
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
LD50 was found to be 15.75 g/kg (11.46 - 21.66 g/kg)
Executive summary:

A 1978 production batch of Kronitex TCP was screened for acute oral toxicity and LD50 was found to be 15.75 g/kg (11.46 - 21.66 g/kg). No classification is applicable.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
15 750 mg/kg bw
Quality of whole database:
K2

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:
16 May 2017 to 05 Jul 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1300 (Acute inhalation toxicity)
Version / remarks:
US EPA OPPTS Guideline 870.1300, Acute Inhalation Toxicity, August, 1998
Deviations:
yes
Remarks:
See "Any other informatino on material and methods inc. tables" below
Qualifier:
according to guideline
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Version / remarks:
OECD Guidelines for the Testing of Chemicals, Section 403, Acute Inhalation Toxicity, September, 2009.
Deviations:
no
GLP compliance:
yes
Test type:
traditional method
Limit test:
no
Specific details on test material used for the study:
A full explanation and analytical report is detailed within Section 13.2 - Material tested for the CoRAP studies
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
Animal Receipt and Acclimation
The Sprague Dawley (Crl:CD[SD]) rats used for this study were received in good health from Charles River Raleigh on 09 May 2017 and 13 Jun 2017. Each animal was inspected by a qualified technician upon receipt. The rats were uniquely identified by ear tags displaying the animal numbers and were acclimated to the laboratory for a minimum of 6 days. During the acclimation period, the rats were observed twice daily for mortality and moribundity.
The animals were subjected to restraint in the nose-only exposure holding tubes for 1 hour on a week day prior to test substance exposure. Following the restraint period, each animal was observed and there were no clinical signs of injury or stress. Animals were held in restraint tubes for at least 15 minutes prior to initiation of exposure after they were delivered to the exposure room.

Animal Housing
Upon arrival, all animals were housed individually in suspended wire-mesh cages. Caging was cleaned at appropriate intervals. The animals were maintained in accordance with Charles River SOPs and the Guide for the Care and Use of Laboratory Animals.1 On the day of exposure, the animals were placed in nose-only exposure holding tubes in the animal room, transported to the exposure room, exposed for the requisite duration, and then returned to their home cages.

Diet, Drinking Water, and Maintenance
The basal diet used in this study, PMI Nutrition International, LLC, Certified Rodent LabDiet® 5002 (block), is a certified feed with appropriate analyses performed by the manufacturer and provided to Charles River Ashland. Reverse osmosis-treated water supplying the facility is analyzed for contaminants according to Charles River SOPs. Filters servicing the automatic watering system are changed regularly according to Charles River SOPs. The results of the diet and water analyses are retained with the Charles River facility data. No contaminants were present in animal feed or water at concentrations sufficient to interfere with the objectives of this study. The basal diet and reverse osmosis-treated water, delivered by an automatic watering system, were provided ad libitum, except during acclimation to the nose-only restraint and during the exposure periods.

Environmental Conditions
The animal room was maintained with controlled temperature, humidity, and fluorescent lighting (12 hours light/12 hours dark). The light status (on or off) was recorded once every 15 minutes. The room temperature and humidity controls were set to maintain conditions of 73°F ± 5°F (approximately 23°C ± 3°C) and 50% ± 20% relative humidity. Room temperature and relative humidity were monitored continuously using the Metasys DDC Electronic Environmental control system and were scheduled for data collection on an hourly basis. Actual mean daily temperature ranged from 71.4°F to 73.5°F (21.9°C to 23.1°C) and mean daily relative humidity ranged from 37.7% to 49.4% during the study. Air handling units were set to provide a minimum of 10 fresh air changes per hour, 100% fresh air.

Enrichment
Each animal was provided devices for environmental enrichment and to aid in maintaining the animals’ oral health (starting during acclimation). Enrichment devices were not available during the restraint acclimation and the exposure periods and were sanitized approximately weekly. Additional enrichment was provided following acclimation to nose-only exposure restraint tubes.

Assignment of Animals to Treatment Groups
Animals used in the study were selected from available stock and assigned to groups using an arbitrary assignment. The animals were selected based on age and body weight requirements and on the appearance of general good health. The selected animals were approximately 8 weeks old at initiation of exposure; body weight values ranged from 296 g to 325 g for males and from 196 g to 236 g for females. Individual body weights at assignment were within ± 20% of the mean for each sex.
Route of administration:
inhalation: aerosol
Type of inhalation exposure:
nose only
Vehicle:
air
Mass median aerodynamic diameter (MMAD):
>= 2.3 - <= 3 µm
Geometric standard deviation (GSD):
>= 1.93 - <= 1.94
Remark on MMAD/GSD:
Two aerosol particle size measurements were conducted during each exposure using a 7-stage cascade impactor (Model No. 02-100-2L-A, In-Tox Products; Moriarty, NM). Pre-weighed, 22-mm stainless steel discs were used for the collection substrates for stages 1 through 7.
A 25-mm glass-fiber filter (Type A/E, PALL Corporation) was used as the collection substrate for the final stage. The sample flow rate was measured using a Mini-Buck Calibrator (Model M-5). Samples were collected at approximately 2.0 LPM for 0.5 and 0.25 minutes for the 1.3 and 5.2 mg/L groups, respectively. The substrates and filter were re-weighed and the particle size was calculated based on the impactor stage cut-offs. The aerosol size was expressed as the mass median aerodynamic diameter (MMAD) and the geometric standard deviation (GSD).
Details on inhalation exposure:
Exposure Methods
Exposures were conducted using a 7.9-L stainless steel, conventional nose-only exposure system with grommets in exposure ports to engage the animal holding tubes (designed and fabricated by Charles River). Animals were restrained in nose-only exposure holding tubes during exposure.
Airflow rates through the system were determined from the airflow required for aerosol generation and the dilution airflow, and provided sufficient volumes for the number of animals to be exposed and for exposure atmosphere sampling. Airflow to the exposure system was provided using a dry, breathing quality, in-house, compressed air source and an in-house supply air source utilizing HEPA filtration and activated charcoal to pre-treat room air.
Exposure atmosphere environmental conditions were recorded at approximately 60-minute intervals during the exposure. A temperature and relative humidity transmitter probe (Model No. HX94C, Omega Engineering, Inc.; Stamford, CT) was used with a display unit to monitor temperature and percent relative humidity.
Oxygen content was measured during the pre-exposure method development phase using a Dräger PAC III equipped with a calibrated oxygen sensor (Draeger Safety, Inc.; Pittsburgh, PA).
The oxygen content was 20.9% for both target concentrations.
The time required to attain 99% of the equilibrium concentration (or clearance time for the concentration to decrease from the equilibrium concentration) was calculated.
The calculated t99 equilibrium time for this study was approximately 1 minute. The start of the 4-hour animal exposure began when animals were placed on the system and the generation was initiated. The exposure period was terminated after 4 hours when the test substance generation was stopped.

Exposure Atmosphere Generation Methods
The system operated as follows. A 3-jet and 6-jet Collison nebulizer (BGI, Inc.; Waltham, MA) was used for the 1.3 and 5.2 mg/L groups, respectively. The test substance was metered at a known flow rate via 1/8-inch Teflon tubing using a Razel syringe pump (Model No. A-99.EJM, Razel Scientific Instruments, Inc.; Stamford, CT) and a 20-mL and 60-mL BD plastic syringe (Becton Dickinson and Co.; Franklin Lakes, NJ) for the 1.3 and 5.2 mg/L groups, respectively. Using a regulator (Model No. 8802K, Coilhose Pneumatics Inc.; East Brunswick, NJ), dry, breathing quality, in-house, compressed air at a controlled pressure was supplied to the air port of the nebulizer to effect atomization of the test substance.
The aerosol was delivered to the nose-only exposure system through 22-mm respiratory tubing.
For the 5.2 mg/L group, the test substance aerosol was directed to a 4.5-L glass chromatography jar where larger particles were removed prior to entering nose-only exposure system. Using a rotameter-type flowmeter (Model No. 127mm, Barnant Co./Gilmont Instruments; Barrington, IL), humidified supply air was added at a tee fitting placed in-line prior to the exposure system. Exhaust atmosphere was filtered using a Solberg filter (Solberg Manufacturing, Inc.; Itasca, IL) prior to entering the facility exhaust system, which consists of redundant exhaust blowers preceded by activated-charcoal and HEPA-filtration units. Negative pressure within the exposure system was checked at an open T-fitting in-line prior to the Solberg filter.
Analytical verification of test atmosphere concentrations:
yes
Remarks:
Actual exposure concentrations were determined approximately every 17 to 45 minutes using standard gravimetric methods.
Duration of exposure:
4 h
Remarks on duration:
As per the test guideline.
Concentrations:
Two range-finding exposures were conducted at mean concentrations of 1.3 and 4.8 mg/L.
For the definitive study, the test substance was administered tat concentrations of 1.3 and 5.2 mg/L.

Nominal Exposure Concentrations
A nominal exposure concentration was calculated from the total amount of test substance used during the generation period and the total volume of air that passed through the exposure system.
The amount of test substance removed using the chromatography jar was not accounted for in the nominal calculation for the 5.2 mg/L group.

Actual Exposure Concentrations
Actual exposure concentrations were determined approximately every 17 to 45 minutes using standard gravimetric methods. Samples were collected on pre-weighed, 25-mm glass-fiber filters (Type A/E, PALL Corporation; Ann Arbor, MI) held in a closed-faced filter holder positioned in the animal exposure port (i.e. in the breathing zone of the animal) of the nose-only exposure system. A measured volume of the exposure atmosphere was pulled through the filter to quantitatively collect aerosol particles. The sample flow rate was controlled using a sapphire critical orifice (Model No. 24, O’Keefe Controls Co.; Trumbull, CT) and measured using a Mini-Buck Calibrator (Model No. M-5, A.P. Buck Inc.; Orlando, FL) prior to each sample.
Samples were collected at approximately 430 mL/minute for 2 and 1 minute for the 1.3 and 5.2 mg/L groups, respectively. Following sample collection, the filters were re-weighed and the concentration calculated as the filter weight difference divided by the sample volume.
No. of animals per sex per dose:
Two range-finding exposures were conducted using 2 rats/sex at mean concentrations of 1.3 and 4.8 mg/L.
For the definitive study, the test substance was administered to 2 groups of 5 male and 5 female Sprague Dawley rats via nose-only inhalation exposure as a liquid droplet aerosol at concentrations of 1.3 and 5.2 mg/L.
Control animals:
no
Details on study design:
The selected route of administration was nose-only exposure methods as a means of reducing the potential for dermal exposure or oral exposure resulting from grooming. The animal model, the albino rat, is generally recognized as appropriate for acute inhalation toxicity studies.

Mortality
All animals were observed for mortality, abnormalities, and signs of pain and distress twice daily for 14 days (once in the morning and once in the afternoon, except on the day of scheduled necropsy).

Clinical Observations
All animals were observed once during the exposure, where only clinical signs visible in nose-only exposure restraint tubes were recorded. All animals were observed for clinical signs of toxicity after the 4-hour exposure when removed from exposure system, once 1 to 2 hours post-exposure, and once daily for 14 days following exposure. Observations were recorded and included, but were not limited to: changes in the skin and fur, eyes and mucous membranes and also changes to the respiratory, circulatory, autonomic and central nervous systems, somatomotor activity, and behavior pattern.

Body Weights
Body weights were obtained prior to exposure on Study Day 0 and on Post-Exposure Days 1, 3, 7, and 14.

Necropsy
Animals at the scheduled necropsy were euthanized by isoflurane anesthesia followed by exsanguination and subject to necropsy. The major organ systems of the cranial, thoracic, and abdominal cavities were examined for all animals. No tissue or organs were retained and the carcasses were discarded after necropsy.
Statistics:
Not specified
Preliminary study:
None of the animals in the range-finding groups died during the study.
Key result
Sex:
male/female
Dose descriptor:
LC0
Effect level:
> 5.2 mg/L air
Based on:
test mat.
Exp. duration:
4 h
Mortality:
None of the animals died during exposure or during the 14-day post-exposure observation period.
Clinical signs:
other: There were no clinical observations noted during the exposure. Following exposure, increased respiration was observed for 1 male in the 5.2 mg/L group and vocalization upon handling was observed for 1 female in the 1.3 mg/L group. At the 1 to 2 hour post-
Body weight:
There were no remarkable changes in body weight in the 1.3 and 5.2 mg/L groups. From Study Day 0 to Study Day 1, 1 male and 2 females in the 1.3 mg/L group lost 2 to 4 grams and 5 males and 4 females in the 5.2 mg/L group lost 3 to 12 grams. All animals surpassed their initial (Study Day 0) body weight by Study Day 14 and were considered normal.
Gross pathology:
Macroscopic findings noted for animals at the scheduled necropsy were enlarged mandibular lymph node for 1 male in the 1.3 mg/L group and 1 male in the 5.2 mg/L group, dark red discoloration of the mandibular lymph node for 1 female in the 5.2 mg/L group and clear fluid contents in the uterus for 2 females in the 5.2 mg/L group. There were no other macroscopic findings for animals at the scheduled necropsy.

Mean Exposure System Environmental Conditions

Group (mg/L):

1.3

5.2

Temperature (°C)

23

21

Standard Deviation:

0.5

1.4

 

 

 

Relative Humidity (%):

40

25

Standard Deviation:

2.5

2.6

 

 

 

Airflow Rate (LPM):

44.7

41.8

Standard Deviation:

1.59

1.73

 

 

 

N:

4

4

 

Nominal Exposure Concentrations

Group (mg/L):

1.3

5.2

Test Substance Used (g):

16.7

68.4

Nominal Concentration (mg/L):

1.6

6.8

 

Mean Actual Exposure Concentrations

Group (mg/L):

1.3

5.2

Target Concentration (mg/L):

1.2

5.0

Mean Concentration (mg/L):

1.3

5.2

Standard Deviation:

0.10

0.34

N:

6

5

 

Mean Particle Size

Group (mg/L):

1.3

5.2

Mean MMAD (microns):

3.0

2.3

Mean GSD:

1.93

1.94

N:

2

2

 

INH.TOX. STUDY OF DISFLAMOLL TKP-P AND KRONITEX TCP IN RATS

SUMMARY OF CLINICAL FINDINGS: TOTAL OCCURRENCE/NO. OF ANIMALS

---- MALE ----

 

DAY 0 POST-EXPOSURE

GROUP:

1

2

BODY / INTEGUMENT

-WET CLEAR MATERIAL DORSAL HEAD

-WET CLEAR MATERIAL FACIAL AREA

-WET CLEAR MATERIAL DORSAL NECK

-WET CLEAR MATERIAL VENTRAL NECK

-WET CLEAR MATERIAL FORELIMB(S)

-WET CLEAR MATERIAL LEFT LATERAL TRUNK

-WET CLEAR MATERIAL RIGHT LATERAL TRUNK

-WET CLEAR MATERIAL VENTRAL TRUNK

-WET CLEAR MATERIAL UROGENITAL AREA

 

1/ 1

3/ 3

0/ 0

0/ 0

1/ 1

2/ 2

1/ 1

5/ 5

0/ 0

 

0/ 0

1/ 1

3/ 3

5/ 5

0/ 0

3/ 3

3/ 3

3/ 3

5/ 5

CARDIO-PULMONARY

-INCREASED RESPIRATION

 

0/ 0

 

1/ 1

EYES/EARS/NOSE

-WET RED MATERIAL AROUND NOSE

-DRIED RED MATERIAL AROUND LEFT EYE

-DRIED RED MATERIAL AROUND RIGHT EYE

 

0/ 0

0/ 0

0/ 0

 

1/ 1

1/ 1

1/ 1

ORAL/DENTAL

-WET CLEAR MATERIAL AROUND MOUTH

-WET RED MATERIAL AROUND MOUTH

 

0/ 0

0/ 0

 

3/ 3

1/ 1

ACUTES

-WET RED MATERIAL VENTRAL NECK

 

1/ 1

 

0/ 0

---- FEMALE ----

 

DAY 0 POST-EXPOSURE

GROUP:

1

2

BEHAVIOR/CNS

-VOCALIZATION UPON HANDLING

 

1/ 1

 

0/ 0

BODY/INTEGUMENT

-WET CLEAR MATERIAL DORSAL HEAD

-WET CLEAR MATERIAL FACIAL AREA

-WET CLEAR MATERIAL DORSAL NECK

-WET CLEAR MATERIAL VENTRAL NECK

-WET CLEAR MATERIAL FORELIMB(S)

-WET CLEAR MATERIAL LEFT LATERAL TRUNK

-WET CLEAR MATERIAL RIGHT LATERAL TRUNK

-WET CLEAR MATERIAL VENTRAL TRUNK

-WET CLEAR MATERIAL DORSAL RUMP

WET-CLEAR MATERIAL UROGENITAL AREA

-WET CLEAR MATERIAL HINDLIMB(S)

 

3/ 3

3/ 3

4/ 4

1/ 1

5/ 5

1/ 1

0/ 0

5/ 5

1/ 1

4/ 4

0/ 0

 

1/ 1

3/ 3

5/ 5

5/ 5

5/ 5

3/ 3

2/ 2

5/ 5

3/ 3

4/ 4

3/ 3

ORAL/DENTAL

-WET CLEAR MATERIAL AROUND MOUTH

 

0/ 0

 

3/ 3

ACUTES

-WET RED MATERIAL FACIAL AREA

 

1/ 1

 

0/ 0

---- MALE ----

TABLE RANGE:

DAY 1 TO DAY 14

GROUP:

1

2

NORMAL

-NO SIGNIFICANT CLINICAL OBSERVATIONS

 

68/ 5

 

68/ 5

DISPOSITION

-SCHEDULED NECROPSY

 

5/ 5

 

5/ 5

DERMAL OBSERVATIONS

-HAIR LOSS FORELIMB(S)

 

2/ 1

 

2/ 1

---- FEMALE ----

TABLE RANGE:

DAY 1 TO DAY 14

GROUP:

1

2

NORMAL

-NO SIGNIFICANT CLINICAL OBSERVATIONS

 

61/ 5

 

64/ 5

DISPOSITION

-SCHEDULES NECROPSY

 

5/ 5

 

5/ 5

BODY INTEGUMENT

-DRIED CLEAR MATERIAL DORSAL HEAD

-DRIED CLEAR MATERIAL DORSAL NECK

-DRIED CLEAR MATERIAL VENTRAL TRUNK

-DRIED CLEAR MATERIAL DORSAL RUMP

-DRIED CLEAR MATERIAL UROGENITAL AREA

 

1/ 1

8/ 3

0/ 0

0/ 0

0/ 0

 

1/ 1

4/ 4

1/ 1

3/ 3

1/ 1

DERMAL OBSERVATIONS

-HAIR LOSS FORELIMB(S)

 

1/ 1

 

0/ 0

1 – 1.3 MG/L    2 – 5.2 MG/L

 

(1-2 HOUR POST-EXPOSURE)

INH.TOX. STUDY OF DISFLAMOLL TKP-P AND KRONITEX TCP IN RATS

SUMMARY OF CLINICAL FINDINGS: TOTAL OCCURRENCE/NO. OF ANIMALS

---- MALE ----

TABLE RANGE:

DAY 0 TO DAY 14

GROUP:

1

2

NORMAL

-NO SIGNIFICANT CLINICAL OBSERVATIONS

 

1/ 1

 

1/ 1

BEHAVIOR/CNS

-VOCALIZATION UPON HANDLING

 

0/ 0

 

2/ 2

BODY / INTEGUMENT

-DRIED CLEAR MATERIAL FACIAL AREA

-DRIED CLEAR MATERIAL DORSAL NECK

-DRIED CLEAR MATERIAL FORELIMB(S)

-DRIED CLEAR MATERIAL LEFT LATERAL TRUNK

-DRIED CLEAR MATERIAL RIGHT LATERAL TRUNK

-DRIED CLEAR MATERIAL VENTRAL TRUNK

-DRIED CLEAR MATERIAL UROGENITAL AREA

 

2/ 2

0/ 0

0/ 0

1/ 1

0/ 0

1/ 1

1/ 1

 

1/ 1

1/ 1

1/ 1

3/ 3

2/ 2

1/ 1

1/ 1

CARDIO-PULMONARY

-RAPID RESPIRATION

 

0/ 0

 

2/ 2

EYES/EARS/NOSE

-DRIED RED MATERIAL AROUND LEFT EYE

-DRIED RED MATERIAL AROUND RIGHT EYE

 

1/ 1

1/ 1

 

0/ 0

0/ 0

---- FEMALE ----

TABLE RANGE:

DAY 0 TO DAT 14

GROUP:

1

2

BEHAVIOR/CNS

-VOCALIZATION UPON HANDLING

 

0/ 0

 

2/ 2

BODY/INTEGUMENT

-DRIED CLEAR MATERIAL DORSAL HEAD

-DRIED CLEAR MATERIAL FACIAL AREA

-DRIED CLEAR MATERIAL DORSAL NECK

-DRIED CLEAR MATERIAL VENTRAL NECK

-DRIED CLEAR MATERIAL FORELIMB(S)

-DRIED CLEAR MATERIAL LEFT LATERAL TRUNK

-DRIED CLEAR MATERIAL RIGHT LATERAL TRUNK

-DRIED CLEAR MATERIAL VENTRAL TRUNK

-DRIED CLEAR MATERIAL DORSAL RUMP

-DRIED CLEAR MATERIAL UROGENITAL AREA

 

2/ 2

2/ 2

4/ 4

1/ 1

2/ 2

0/ 0

0/ 0

4/ 4

0/ 0

3/ 3

 

0/ 0

2/ 2

3/ 3

2/ 2

1/ 1

1/ 1

2/ 2

5/ 5

3/ 3

2/ 2

CARDIO-PULMONARY

-RAPID RESPIRATION

 

0/ 0

 

1/ 1

1 – 1.3 MG/L    2 – 5.2 MG/L

 

INH.TOX. STUDY OF DISFLAMOLL TKP-P AND KRONITEX TCP IN RATS

SUMMARY OF BODY WEIGHTS [G]

MALES

GROUP:

1.3 MG/L

5.2 MG/L

DAY

0

 

MEAN

S.D.

N

 

302.

7.2

5

 

314.

6.0

5

1

 

MEAN

S.D.

N

 

304.

6.3

5

 

307.

6.9

5

3

 

MEAN

S.D.

N

 

319.

4.1

5

 

326.

10.3

5

7

 

MEAN

S.D.

N

 

342.

5.7

5

 

359.

10.0

5

14

 

MEAN

S.D.

N

 

389.

11.9

5

 

409.

10.9

5

FEMALES

GROUP:

1.3 MG/L

5.2 MG/L

DAY

0

 

MEAN

S.D.

N

 

203.

6.6

5

 

227.

6.2

5

1

 

MEAN

S.D.

N

 

205.

5.4

5

 

223.

9.3

5

3

 

MEAN

S.D.

N

 

212.

7.5

5

 

230.

10.4

5

7

 

MEAN

S.D.

N

 

220.

9.6

5

 

236.

10.2

5

14

 

MEAN

S.D.

N

 

235.

14.1

5

 

256.

9.0

5

 

INH.TOX. STUDY OF DISFLAMOLL TKP-P AND KRONITEX TCP IN RATS

SUMMARY OF BODY WEIGHT CHANGES [G]

MALES

GROUP:

1.3 MG/L

5.2 MG/L

DAY

0 TO 1

 

 

 

MEAN

S.D.

N

1.

3.1

5

-8.

2.4

5

1 TO 3

 

 

 

MEAN

S.D.

N

15.

3.2

5

19.

3.5

5

3 TO 7

 

 

 

MEAN

S.D.

N

24.

1.8

5

33.

3.9

5

7 TO 14

 

 

 

MEAN

S.D.

N

46.

7.5

5

50.

6.4

5

FEMALES

GROUP:

1.3 MG/L

5.2 MG/L

DAY

0 TO 1

 

 

 

MEAN

S.D.

N

2.

5.7

5

-4.

5.0

5

1 TO 3

 

 

 

MEAN

S.D.

N

7.

5.9

5

7.

4.7

5

3 TO 7

 

 

 

MEAN

S.D.

N

8.

2.6

5

6.

3.8

5

7 TO 14

 

 

 

MEAN

S.D.

N

15.

8.1

5

20.

3.5

5

 

INH.TOX. STUDY OF DISFLAMOLL TKP-P AND KRONITEX TCP IN RATS

SUMMARY OF MACROSCOPIC FINDINGS

---- MALE ----

GROUP:

1

2

NUMBER OF ANIMALS IN DOSE GROUP

NUMBER OF ANIMALS TERMINALLY EUTHANIZED

5

5

5

5

MANDIBULAR, LN

-ENLARGED

 

1

 

1

NO SIGNIFICANT CHANGES OBSERVED – ALL EXAMINED TISSUES

4

4

---- FEMALE ----

GROUP:

1

2

NUMBER OF ANIMALS IN DOSE GROUP

NUMBER OF ANIMALS TERMINALLY EUTHANIZED

5

5

5

5

MANDIBULAR, LN

-DISCOLORATION, DARK RED

 

0

 

1

UTERUS

-CONTENTS, CLEAR FLUID

 

0

 

2

NO SIGNIFICANT CHANGES OBSERVED – ALL EXAMINED TISSUES

5

2

1 – 1.3 MG/L    2 - 5.2 MG/L

Deviations

A GLP deviation from 40 CFR Sections 160.130( e) and 792.130( e) occurred.

The individual inputting data in an automated data collection system was not identified at the time of that data input. On 18 May 2017, Operator No. 1122 collected data using the electronic log-in for Operator No. 247. The data was still recorded, but not by the person logged into the computer system. Therefore, there was no impact on the collected study data.

Interpretation of results:
GHS criteria not met
Conclusions:
Based on the results of this study, the LC50 of Disflamoll TKP-P and Kronitex TCP (1:1 Mixture) was > 5.2 mg/L when male and female Crl:CD(SD) rats were exposed to an aerosol of the test substance as a single, 4-hour, nose-only exposure.
Executive summary:

SUMMARY

Objective

The objective of this study was to determine the acute inhalation toxicity of Disflamoll TKP-P and Kronitex TCP (1:1 Mixture) when administered as a single, 4-hour, nose-only inhalation exposure to rats.

 

Test Guidelines

The protocol was designed in accordance with the US EPA OPPTS Guideline 870.1300, Acute Inhalation Toxicity, August, 1998 and the OECD Guidelines for the Testing of Chemicals, Section 403, Acute Inhalation Toxicity, September, 2009.

 

Study Design

The acute inhalation toxicity of Disflamoll TKP-P and Kronitex TCP (1:1 Mixture) was evaluated in this 4-hour, single-exposure study in rats. Two range-finding exposures were conducted using 2 rats/sex at mean concentrations of 1.3 and 4.8 mg/L. For the definitive study, the test substance was administered to 2 groups of 5 male and 5 female Sprague Dawley rats via nose-only inhalation exposure as a liquid droplet aerosol at concentrations of 1.3 and 5.2 mg/L. Mean mass median aerodynamic diameter ± the geometric standard deviation were 3.0 ± 1.93 and 2.3 ± 1.94 μm for the 1.3 and 5.2 mg/L groups, respectively.

Mortality, clinical observations, body weights, and body weight changes were evaluated over a 14-day post-exposure observation period. Necropsies were conducted on all animals.

 

Results

None of the animals in the range-finding nor the definitive groups died during the study.

Significant clinical observations following exposure included increased respiration for the 5.2 mg/L group and vocalization upon handling for the 1.3 mg/L group. Significant clinical observations at the 1 to 2 hour post-exposure period included rapid respiration and vocalization upon handling for the 5.2 mg/L group. There were no significant clinical observations during the 14-day post-exposure observation period. All animals were considered clinically normal by Study Day 1.

One male and 2 females in the 1.3 mg/L group and 5 males and 4 females in the 5.2 mg/L group lost weight from Study Day 0 to Study Day 1. All animals surpassed their initial (Study Day 0) body weight by Study Day 14.

Macroscopic findings noted for animals at the scheduled necropsy were enlarged mandibular lymph node for the 1.3 and 5.2 mg/L groups and dark red discoloration of the mandibular lymph node and clear fluid contents in the uterus for the 5.2 mg/L group. There were no other macroscopic findings for animals at the scheduled necropsy.

 

Conclusions

Based on the results of this study, the LC50 of Disflamoll TKP-P and Kronitex TCP (1:1 Mixture) was > 5.2 mg/L when male and female Sprague Dawley rats were exposed to an aerosol of the test substance as a single, 4-hour, nose-only exposure.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LC50
Value:
5.2 mg/m³
Quality of whole database:
1

Acute toxicity: via dermal route

Link to relevant study records
Reference
Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
January 4 1979 to February 22 1979
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Non-GLP study, no specific methodology followed although fully documented report available.
Qualifier:
no guideline followed
Principles of method if other than guideline:
Acute dermal toxicity in rabbits was determined according to the procedures suggested by Hagan. New Zealand white rabbits were assigned to groups and maintained under standard laboratory conditions prior to administration of the test material.

Prior to determination of LD50 , single animals were dosed at various levels to determine a range within which the LD50 could be determined. The animals were prepared by clipping the skin of the mid-dorsal area of the trunk free of hair. Animals were further prepared by introducing longitudinal epidermal incisions over the clipped skin surface, thus enhancing the penetrability of the stratum corneum. Applications were made under gauze patches. The dosed area was then covered with an impermeable p lastic wrapping for 24 hours, after which it was removed and the skin gently cleansed.

Animals were returned to quarters where food and water were available ad libitum following the dose application, and were observed fourteen full days following application to determine mortality during this period.

Following the range finding, rabbits were divided into groups of six , sexes equally distributed, one-half of the animals abraded. Groups were dosed, as described above, at graded dose levels according to the results of the range finding previously performed.

Animals were observed for signs of pharmacologic activity and drug toxicity at 1, 3, 6, and 24 hours post-dosage. Observations were made daily thereafter for a total of fourteen days. Non-surviviors and animals sacrificed a t the end of the 14 day observation period were subjected to complete gross necropsy. LD50 together with 95% confidence limits was determined, where possible, by the method of Litchfield and Wilcoxin.

GLP compliance:
no
Test type:
standard acute method
Limit test:
no
Species:
rabbit
Strain:
New Zealand White
Sex:
male/female
Details on test animals or test system and environmental conditions:
All animals are fed and watered ad libitum; with Wayne animal feeds used exclusively. Animals are received from Summit View Farm, Belvidere, New
Jersey and are conditioned prior to use.
Rabbits were divided into groups of six , sexes equally distributed, one-half of the animals abraded.
Type of coverage:
occlusive
Vehicle:
not specified
Remarks:
Presumably neat.
Details on dermal exposure:
The animals were prepared by clipping the skin of the mid-dorsal area of the trunk free of hair. Animals were further prepared by introducing longitudinal epidermal incisions over the clipped skin surface, thus enhancing the penetrability of the stratum corneum. Applications were made under
gauze patches. The dosed area was then covered with an impermeable plastic wrapping for 24 hours, after which it was removed and the skin gently cleansed.
Duration of exposure:
24 h
Doses:
Single doses, at 3.14, 5, 6.3, 10, 12.6 g/kg
No. of animals per sex per dose:
3
Control animals:
no
Details on study design:
Acute dermal toxicity in rabbits was determined according to the procedures suggested by Hagan (1959). New Zealand white rabbits were assigned to groups and maintained under standard laboratory conditions prior to administration of the test material.
Prior to determination of LD50 single animals ere dosed at various levels to determine a range within which the LD could be determined.
The animals were prepared by clipping the skin of the mid-dorsal area of the trunk free of hair. Animals were further prepared by introducing longitudinal epidermal incisions over the clipped skin surface, thus enhancing the penetrability of the stratum corneum. Applications were made under
gauze patches. The dosed area was then covered with an impermeable plastic wrapping for 24 hours, after which it was removed and the skin gently cleansed. Animals were returned to quarters where food and water were available ad libitum following the dose application, and were observed fourteen full days following application to determine mortality during this period.
Following the range finding, rabbits were divided into groups of six , sexes equally distributed, one-half of the animals abraded. Groups were dosed, as described above, at graded dose levels according to the results of the range finding previously performed. Animals were observed for signs of pharmacologic activity and drug toxicity at 1, 3, 6, and 24 hours post-dosage. Observations were made daily thereafter for a total of fourteen days. Non-survivors and animals sacrificed at the end of the 14 day observation period were subjected to complete gross necropsy. LD50 together with 95% confidence limits was determined where possible, by the method of Litchfield and Wilcoxin (1949).
Statistics:
None specified.
Preliminary study:
Not applicable.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
3.7 other: g/kg
Based on:
test mat.
95% CL:
2.31 - 5.92
Mortality:
Range finding study: 2 animals at 10.0 and 15.0 g/kg dose levels were found dead on Day 2.

Full study:
Dose level: 3.14 g/kg: No deaths
Dose level: 5.00 g/kg: 2 deaths were noted on Day 3. 1 death occured on Day 5.
Dose level: 6.30 g/kg: 2 deaths were noted on Day 4. 1 death occured on Day 5. Animal 6 died on Day 10 and was replaced.
Dose level: 10.00 g/kg: Deaths were noted on Days 2,3,4,5 and Day 9 (unspecified)
Dose level: 12.60 g/kg: 1 death was noted on Day 2; 2 deaths were noted on Day 3. 2 deaths were noted on Day 4.
Clinical signs:
Dose level: 3.14 g/kg: Skin pliable and non - irritated. No gross changes observed. Animal 3, on days 4 - 6: Possible respiratory infection - slight difficulty breathing.

Dose level: 5.00 g/kg: Skin pliable and non - irritated ; faeces loose. No gross changes observed

Dose level: 6.30 g/kg: Skin pliable and non - irritated. Animal 1 Sacrificed on Day 4; moribund. Hair matted and unkempt; faeces loose

Dose level: 10.00 g/kg: Skin pliable and non - irritated ; faeces loose. Hair matted and unkempt. No gross internal changes observed.

Dose level: 12.60 g/kg: Skin moderately reddened. Hair matted. No gross internal changes observed. Animals 5 and 6 sacrificed on Day 4

Body weight:
With the exception of dose level 3.14 g/kg; all animals demonstrated bodyweight loss during the course of the study.
Gross pathology:
No pathological findings were recorded.

Table 1

 

Dose level g/kg

Sex

Dead/dosed

% Mortality

Range finding

1.00

1M

0/1

0

5.00

1M

0/1

0

10.00

1F

1/1

100

15.00

1F

1/1

100

Test dosage

3.14

3M:3F

0/3:0/3

0

5.00

3M:3F

2/3:2/3

67

6.30

3M:3F

2/3:2/3

67

10.00

3M:3F

3/3:2/3

83

 

LD50 = 3.7 (2.31 - 5.92) g/kg

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The acute dermal toxicity of Tricresyl Phosphate (LD50) is 3.7 (2.31 - 5.92) g/kg
Executive summary:

The study has shown the acute dermal toxicity of Tricresyl Phosphate (LD50) to be 3.7 (2.31 - 5.92) g/kg. No classification is applicable.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
3 700 mg/kg bw

Additional information

Testing on the above endpoints gave the following results:

 

Acute toxicity: Oral.

 

2 main studies are available for the derivation of LD50 acute oral toxicity as follows:

 

LD 50: 15750 mg/kg

 

LD50: >20000 mg/kg

 

Both studies conducted at different times show equivalent toxicological profiles and values; hence it is

considered that applicable of oral LD50 values results in no classification. Other studies offered as supporting information are not considered relevant given that definitive values have been obtained in these two studies.

 

Acute toxicity: Dermal.

 

Two main studies are presented for this endpoint as follows:

 

LD 50: 3700 mg/kg

 

LD50: >10000 mg/kg

 

Both studies conducted at different times show equivalent toxicological profiles and values; hence it is considered that applicable of oral LD50 values results in no classification. Other studies offered as supporting information are not considered relevant given that definitive values have been obtained in these two studies.

 

Acute toxicity: Inhalation.

 

Three studies are again presented for this endpoint as follows:

Key:

LC50 (4 hour) > > 5.2 mg/L (air) 

Supporting:

LC50 (1-hour): >11.1 mg/l (air)

LC 50 (1-hour) < 200 mg/l (air)

 

Key:

None of the animals in the range-finding nor the definitive groups died during the study.

Significant clinical observations following exposure included increased respiration for the 5.2 mg/L group and vocalization upon handling for the 1.3 mg/L group. Significant clinical observations at the 1 to 2 hour post-exposure period included rapid respiration and vocalization upon handling for the 5.2 mg/L group. There were no significant clinical observations during the 14-day post-exposure observation period. All animals were considered clinically normal by Study Day 1.

One male and 2 females in the 1.3 mg/L group and 5 males and 4 females in the 5.2 mg/L group lost weight from Study Day 0 to Study Day 1. All animals surpassed their initial (Study Day 0) body weight by Study Day 14.

Macroscopic findings noted for animals at the scheduled necropsy were enlarged mandibular lymph node for the 1.3 and 5.2 mg/L groups and dark red discoloration of the mandibular lymph node and clear fluid contents in the uterus for the 5.2 mg/L group. There were no other macroscopic findings for animals at the scheduled necropsy.

Based on the results of this study, the LC50 of Disflamoll TKP-P and Kronitex TCP (1:1 Mixture) was > 5.2 mg/L when male and female Sprague Dawley rats were exposed to an aerosol of the test substance as a single, 4-hour, nose-only exposure.

 

Supporting:

Neither study in isolation is suitable for hazard assessment for this endpoint; however given the low volatility of the substance, extensive exposure by inhalation is not anticipated. There is the potential for exposure by inhalation from some of the categories of use; however as these are as a result of the substance as a component of a product at low level, plus the fact that PPE during use of such products is recommended, it is considered that exposure by inhalation will not pose a hazard. In addition, the dermal route of exposure is considered more appropriate for exposure to the substance, given the nature and conditions of usage.

 

Justification for selection of acute toxicity – oral endpoint

Lower of the two values deemed reliable.

 

Justification for selection of acute toxicity – inhalation endpoint

> limit value achieved.

 

Justification for selection of acute toxicity – dermal endpoint

Lower of the two values deemed reliable.

Justification for classification or non-classification

The above studies have all been ranked reliability 1,2 or 3 according to the Klimish et al system. This ranking was deemed appropriate because the studies were not conducted to GLP or in compliance with agreed protocols. The reports do not detail a specific method; however it documents dose levels and responses in detail, so is deemed appropriate for use in the support of a formal registration. Sufficient dose ranges and numbers are detailed; hence it is appropriate for use based on reliability and animal welfare grounds.

Justification for classification or non classification

The above results triggered no classification under the CLP Regulation (EC No 1272/2008). No classification for acute effects is therefore required.

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