Octadecyl stearate

Endpoint summary

Description of key information

Repeated dose toxicity: Oral

The No Observed Adverse Effect Level (NOAEL) of the test chemical in the rat via oral route of exposure is considered to be 1000 mg/kg body weight in male and female animals.

 

Repeated dose toxicity: Inhalation

Test chemical has very low vapor pressure of 2.16E-09 Pa. (1.62013e-11 mm Hg), so the potential for the generation of inhalable vapours is very low, also the normal conditions of use of this substance will not result in aerosols, particles or droplets of an inhalable size, so exposure to humans via the inhalatory route will be highly unlikely and therefore this end point for repeated dose toxicity by inhalation route of exposure was considered for waiver.

 

Repeated dose toxicity: Dermal

The acute dermal toxicity value for test chemical (as provided in section 7.2.3) is >2000 mg/kg body weight. Given the use of the chemical; repeated exposure by the dermal route is unlikely since the use of gloves is common practice in industries. Thus, it is expected that test chemical shall not exhibit 28 day repeated dose toxicity by the dermal route. In addition, there is no data available that suggests that test chemical shall exhibit repeated dose toxicity by the dermal route. Hence this end point was considered for waiver.

Key value for chemical safety assessment

Toxic effect type:

dose-dependent

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Endpoint:

repeated dose toxicity: oral, other

Type of information:

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

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

experimental data of read across substances

Justification for type of information:

Data for the target chemical is summarized based on the structurally similar read across chemicals.

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

other: as mentioned below

Principles of method if other than guideline:

WoE for the target CAS is summarized based on data from various test chemicals.

GLP compliance:

not specified

Limit test:

no

Species:

rat

Strain:

other: 2. Crj: CD(SD) 3. Crj: CD(SD) 4. No data

Details on species / strain selection:

No data available

Sex:

male/female

Details on test animals or test system and environmental conditions:

2.TEST ANIMALS
- Source: Charles River Japan
- Age at study initiation: 8 week old
- Weight at study initiation:
Male: 312.1 to 363.7 g
Female: 205.3 to 230.8 g
- Fasting period before study: No data
- Housing: Each animal was housed individually in a conditioned room in Japan cage with wire mesh floor cage. Pregnant female rats were bred in cages with pulp and papermade chips as bedding
- Diet (e.g. ad libitum): Solid feed (CE2, Japan Clea) ad libitum
- Water (e.g. ad libitum): Tap water ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 24 ± 1 ° C
- Humidity (%): 50 to 65%
- Air changes (per hr): 15 times / hour
- Photoperiod (hrs dark / hrs light): 12 hours (7 am to 7 pm)

IN-LIFE DATES: From: To: No data

3.No data available

Route of administration:

other: 2.oral: gavage 3.oral: gavage 4. oral: feed

Details on route of administration:

No data available

Vehicle:

corn oil

Details on oral exposure:

2.PREPARATION OF DOSING SOLUTIONS:Test chemical was prepared by suspension in corn oil at dose levels of 0, 100, 300 and 1000 mg/kg. Dosing specimens were prepared at least once weekly

DIET PREPARATION
- Rate of preparation of diet (frequency): No data
- Mixing appropriate amounts with (Type of food): No data
- Storage temperature of food: No data

VEHICLE
- Justification for use and choice of vehicle (if other than water): Corn oil
- Concentration in vehicle: 0, 100, 300 and 1000 mg/kg
- Amount of vehicle (if gavage): 5 mL/Kg bw
- Lot/batch no. (if required): Lot No. V6H2050
- Purity: No data

3. PREPARATION OF DOSING SOLUTIONS: The test chemical was dissolved in corn oil at dose levels of 0, 100, 300 or 1000 mg/kg/day

DIET PREPARATION
- Rate of preparation of diet (frequency): No data available
- Mixing appropriate amounts with (Type of food): No data available
- Storage temperature of food: No data available

VEHICLE
- Justification for use and choice of vehicle (if other than water): Corn oil
- Concentration in vehicle: 0, 100, 300 or 1000 mg/kg/day
- Amount of vehicle (if gavage): No data available
- Lot/batch no. (if required): No data available
- Purity: No data available

4. PREPARATION OF DOSING SOLUTIONS: The test chemical was mixed with feed at dose level of 0, 0.5, 1.0, or 3.0 % (0, 250, 500 or 1500 mg/Kg/day)

DIET PREPARATION
- Rate of preparation of diet (frequency): No data
- Mixing appropriate amounts with (Type of food): No data
- Storage temperature of food: No data

VEHICLE
- Justification for use and choice of vehicle (if other than water): Feed
- Concentration in vehicle: 0, 0.5, 1.0, or 3.0 % (0, 250, 500 or 1500 mg/Kg/day)
- Amount of vehicle (if gavage): No data
- Lot/batch no. (if required): No data
- Purity: No data

Analytical verification of doses or concentrations:

not specified

Details on analytical verification of doses or concentrations:

No data available

Duration of treatment / exposure:

2.Males: 43 days; Females: approx 63 days
3. Males: 42 days Females: From 14 days before mating to day 4 of lactationSatellite group (females): 42 days
4. 2 years

Frequency of treatment:

2.Daily
3. Daily for 42 days for males and daily from 14 days before mating to day 4 of lactation for females
4. Daily

Remarks:

2. Dose:0,100,300,1000 mg/kg bw/day

Remarks:

3. Doses/Concentrations: 0, 100, 300 or 1000 mg/kg/day

Remarks:

4. Dose: 0, 0.5, 1.0, or 3.0 % (0, 250, 500 or 1500 mg/Kg/day)

No. of animals per sex per dose:

2.Total: 104
0 mg/kg/day: 13 males and 13 females
100 mg/kg/day: 13 males and 13 females
300 mg/kg/day: 13 males and 13 females
1000 mg/kg/day: 13 males and 13 females
1. Total: 106 rats

Test group:
0 mg/kg/day: 12 males.12 females
100 mg/kg/day: 12 males, 12 females
300 mg/kg/day: 12 males, 12 females
1000 mg/kg/day: 12 males, 12 females

Satellite group for recovery of females:
0 mg/kg/day: 5
1000 mg/kg/day: 5

3.Total: 106 rats

Test group:

0 mg/kg/day: 12 males.12 females
100 mg/kg/day: 12 males, 12 females
300 mg/kg/day: 12 males, 12 females
1000 mg/kg/day: 12 males, 12 females

Satellite group for recovery of females:
0 mg/kg/day: 5
1000 mg/kg/day: 5


3. Total: 80 males
0 mg/Kg/day: 20 male rats
250 mg/Kg/day: 20 male rats
500 mg/Kg/day: 20 male rats
1500 mg/Kg/day: 20 male rats

Control animals:

yes, concurrent vehicle

Details on study design:

2.- Dose selection rationale: 14 days preliminary repeated dose study was performed. No signs of toxicity were observed even in the group administered with the critical dose of 1000 mg/kg. Hence in the main study, 1000 mg/kg was set at the highest dose, and divided by the tolerance of about 3 to make the medium dose 300 mg/kg and the low dose 100 mg/kg. Corn oil used as a medium for test chemical was administered to the rats of the control group under the same conditions as the test chemical administration group.

- Rationale for animal assignment (if not random): Both males and females were grouped by random weighted stratified random extraction method based on the body weight of the administration start date (8 weeks old), and 13 male and 13 female each were placed in each group.

- Rationale for selecting satellite groups: No data
- Post-exposure recovery period in satellite groups: No data
- Section schedule rationale (if not random): No data

3.1) Parent animals

(1) General state observation
　Throughout the administration period and subsequent 14-day recovery period, animals were observed daily for life and death, appearance, and behavior.

(2) Detailed clinical observation
　The day before the start of administration and once a week thereafter, when the animal is removed from the cage and when it is removed from the cage in an aluminum open field (370 W x 560 D x 40 H mm) outside the cage Easiness, body tension (relaxation to tonicity), skin (color), fur, napping, eye secretions, closed eyelids, eyeball protrusion, lacrimation, nasal discharge (dirt), salivation, lower abdominal coat urine Contamination due to stool, stool around the anus, vocalization, breathing, posture, convulsions, tremor, exploratory behavior (wakefulness), vigilance, spontaneous movement (activity), walking (staggered), abnormal behavior (self-biting, backward facing We observed 29 items such as walking), regularity (excessive hair repair, repetitive turning, etc.), consciousness disorder (confused, catalepsy, coma), limb muscle tone, urination and defecation.

(3) Sensory reflex function test
　Males, males once in the nursing period, males and females in the satellite group, auditory response, visual response, tactile response, pinna reflex, pain response, pupillary reflex, Ipsilateral flexor reflexes, eyelid reflexes, and forward reflexes were examined.

(4) Landing leg width, grip strength and spontaneous momentum measurement
　Males are treated for 41 days, females are treated once during the nursing period, and males and females in the satellite group are on the last treatment day and recovery on the 13th day. The width of the landing leg is measured (measures the width of the footprint when dropped from a height of 30 cm) , Grip strength of forelimbs and hind limbs (rat / mouse grip strength measurement device, MK-380R / FR, Muromachi machine) and males for 30 and 60 minutes, females for 30 minutes (spontaneous momentum measurement device, SUPERMEX, Muromachi machine) , Measured the number of movements in 60 minutes between compartments in the measuring device). Spontaneous momentum of females in the satellite group was measured for 60 minutes.

(5) Body weight and food consumption measurement
　Body weight was measured once a week and on the day of sacrifice. The amount of food consumption was measured once a week for one day (24 hours).

(6) Sexual cycle inspection
　For females, vaginal plaque smears were prepared by Giemsa staining until mating was confirmed following the habituation and quarantine period, and the sexual cycle stage was determined by microscopic examination. The mean sexual cycle was defined as the average number of days in which the late estrus I, in which only keratinocytes were scattered or agglomerated, returned.

(7) Mating and observation of labor conditions
　On the afternoon of the 15th day of administration, females in the same group were placed in male cages (one-to-one) and allowed to live together for up to 14 days until mating was confirmed. Mating was confirmed every morning at a certain time, and the day when sperm was confirmed in the vaginal plug formation or vaginal plaque was defined as day 0 of pregnancy. The state of parturition was also observed at the same time, and the day when the end of parturition was confirmed for each abdomen was defined as day 0 of nursing. From the observation results of mating and delivery, the mating rate (%) [(number of mating animals / number of live animals) × 100], conception rate (%) [(number of conceived females / number of mating females) × 100] and The birth rate (%) [(number of live births / number of pregnant females) x 100] and the duration of pregnancy were calculated for the cases in which delivery was confirmed (days from the 0th day of pregnancy to the day on which delivery was confirmed). The female satellite group did not mate.

(8) Urinalysis
　For males, on the 40th day of administration and on the 8th day of recovery in the satellite group, the animals are placed in metabolic cages for about 3 hours. The resulting urine is used to observe the appearance, test paper method [Multistics, Bayer Sankyo ] Qualitative examination of pH, occult blood, protein, sugar, ketone body, bilirubin and urobilinogen, and examination of sediment (URI-CELL solution, stained with Cambridge Chemical Products Co., Ltd.). Furthermore, urine volume, specific gravity (refractometer, Elmer Optics) and sodium and potassium (electrolyte automatic analyzer, NAKL-132, Toa Denki Kogyo) were measured for urine obtained after 18-hour storage.

(9) Hematology test
　Blood samples were collected from the abdominal aorta under ether anesthesia the day after the administration and recovery period. The animals were weeded from 5 pm the day before blood collection and were given water only. The collected blood is divided into 3 parts, and a part of it is treated with EDTA-2K to prevent coagulation, and with a multi-item automatic blood cell counter (E-4000, Sysmex), the red blood cell count (electric resistance detection method), hemoglobin amount (lauryl) Sodium sulfate-hemoglobin method), hematocrit value (pulse detection method), mean red blood cell volume (MCV), mean red blood cell pigment content (MCH), mean red blood cell pigment concentration (MCHC, calculated above), white blood cell count and platelet count (above, Electrical resistance detection method), smear specimens were prepared, and the reticulocyte count (microscopically stained with Brilliant cresyl blue) and white blood cell percentage (microscopically stained with May-Giemsa) were measured. In some cases, plasma is separated by clotting inhibition treatment with 3.8% sodium citrate solution, and the prothrombin time (PT, Quick one-step method) and activation part are measured using an automatic blood coagulation analyzer (KC-10A, Amerung). Thromboplastin time (APTT, ellagic acid activation method) was measured.

(10) Blood biochemistry test
　Serum is separated from a portion of the collected blood and analyzed using a biochemical automatic analyzer [JCA-BM8, JEOL] with total protein (Biuret method), albumin (BCG method), A / G ratio (calculated value), glucose , Triglyceride, total cholesterol (above, enzymatic method), total bilirubin (diazo method), urea nitrogen (Urease-UV method), creatinine (Jaff), AST, ALT, g-GTP, ALP (above, JSCC method) , LDH (SFBC method), cholinesterase (BTC-DTNB method), calcium (OCPC method) and inorganic phosphorus (enzymatic method), and sodium, potassium and chlorine using an automatic electrolyte analyzer [NAKL-132, Toa Denpa Kogyo Co., Ltd.] (Ion electrode method) was measured.

(11) Pathological examination
　For male slaughtered animals, the day after the 42nd day of administration, in the case of females who were delivered and nurturing well, 5 days were nurtured, and those who were not mated were 24 days after the mating period (the day after the 52nd day). In cases where mating was established but no delivery was observed until 4 days after the scheduled delivery, and in the satellite group on the 14th day after recovery, the blood was sacrificed under ether anesthesia, and the body surface, opening The mucosa and internal organs were observed macroscopically. In addition, weigh (absolute weight) the liver, kidney, adrenal gland, thymus, spleen, brain, heart, pituitary gland, thyroid gland and seminal vesicle and all male testes and epididymis of each male and female. Based on the body weight, the ratio to body weight (relative weight) was calculated. For females, the number of ovaries of the ovaries and the number of uterus implantations were examined, and the implantation rate (%) [(implantation number / luteal number) × 100] was calculated.

　For all cases, brain, pituitary gland, thyroid, thymus, trachea / lung (fixed after infusion), stomach, intestine, heart, liver, spleen, kidney, adrenal gland, bladder, testis, epididymis, prostate, seminal vesicle , Ovary, uterus, spinal cord (neck, chest, lumbar region), sciatic nerve, bone marrow (femur), lymph node (cervical lymph node, mesenteric lymph node), mammary gland, and other grossly abnormal sites, The cells were fixed and preserved with 10% neutral phosphate buffered formalin solution (testis and epididymis were pre-fixed with Bouin's solution). Histopathological examinations were performed on these preserved organs in both sexes in the control and 250 mg / kg groups. All cases were examined for gross abnormalities in each group. In addition, for males and females who were not mated and females who were mated but were not delivered until 4 days after delivery, the testes, epididymis, prostate, and seminal vesicles were used in males, and the ovaries, uterus, and pituitary in females. Were inspected. In the examination, paraffin sections were prepared according to a conventional method, followed by HE staining and microscopic examination. The testis of the control group and the 1000 mg / kg group were subjected to PAS staining and spermatogenesis cycle tests (stages II, III, V, VII and XII).

2) Newborn items
(1) Number of babies, sex ratio and outer surface observation

　After confirming the completion of labor, the number of babies in each abdomen (the total number of babies born and dead) was examined. Gender was determined and sex ratio for each group was calculated. Newborns were observed for abnormalities in the outer surface including the oral cavity.　

(2) General state observation

　Check the general condition and life and death every day. I asked.

(3) Body weight measurement

　For newborns, the total body weight of each abdomen was measured for both sexes on days 0 and 4 of breastfeeding, and the average body weight per animal was calculated.

(4) Pathological examination

　In each case of death, the surviving cases were exsanguinated under ether anesthesia on the fourth day of nursing and the major organs in the thoracoabdominal region were visually observed.

Positive control:

No data available

Observations and examinations performed and frequency:

2.CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Once daily
- Cage side observations :All cases were observed at least once daily during the breeding period.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: observed at least once a day.

BODY WEIGHT: Yes
- Time schedule for examinations: For males on day 1, 8 and 15 of administration and for copulated females on day 1, 8, 15, 22, 29, 36 and 42 days and post menopausal days. For non copulated females the body weight was also measured on the 22nd day of administration. Also, mating females at 0, 7, 14, and 20 days of gestation, females delivered at 0 and 4 days of nursing (autopsy days), females who did not deliver, weighed 25 days pregnant (day of autopsy).

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Time schedule for examinations: For males on day 1, 8 and 15 of administration and for copulated females on day 1, 8, 15, 22, 29, 36 and 42 days and post menopausal days. No food intake was measured during the 2 week mating period. For mating females, food intake was measured between 0-7, 7-14, 14-20 days of pregnancy, and 0-4 days of feeding in females delivered. In addition, feed consumption of mothers who died in the middle of nursing on 04 days of nursing were excluded from the evaluation.
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data

FOOD EFFICIENCY: No
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: Not examined

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Not examined

OPHTHALMOSCOPIC EXAMINATION: Not examined

HAEMATOLOGY: Yes
- Time schedule for collection of blood: At the end of administration period.
- Anaesthetic used for blood collection: Yes, pentobarbital sodium anesthesia
- Animals fasted: No data available
- How many animals: All male animals
- Parameters checked in table [No.?] were examined: No data available
Males: RBC, hemoglobin, hematocrit, MCV, MCH, MCHC platelet, WBC, band neurophil, segmented neurophil, eosinophile, basophile, monocyte and lymphocyte were examined.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: At the end of administration period.
- Animals fasted: Yes, pentobarbital sodium anesthesia
- How many animals: All male animals
- Parameters checked in table [No.?] were examined.:No data available
Males: Total protein, albumin, A/G, BUN, creatinine, glucose, total cholesterol, total bilirubin, triglyceride Na, K, Cl, Ca, inorganic phosphorus, ALP, GPT,GOT, γ-GTP and LDH were examined.

URINALYSIS: No data
- Time schedule for collection of urine:
- Metabolism cages used for collection of urine: No data
- Animals fasted: No data
- Parameters checked in table [No.?] were examined. No data

NEUROBEHAVIOURAL EXAMINATION: No data
- Time schedule for examinations: No data
- Dose groups that were examined: No data
- Battery of functions tested: sensory activity / grip strength / motor activity / other: No data

OTHER:
A. Calculation of the number of births: The number of births (surviving infants + dead children) was examined on day 0 of nursing and the rate of
delivery and the birth rate of babies was obtained. The presence and sex of external birth defects in babies, and calculated the sex ratio of the survivors was also determined.

B. Calculation of the number of deceased children: The number of deceased children was checked every day and the birth rate and neonatal survival rate was noted on 4th day of feeding

C. Weight measurement: Body weight (litter weight) was measured for each sex on a 0 day and a 4 day nursing, and the average body weight per animal was obtained for each abdomen.

D. Necropsy: The surviving infant was mortalized in all the cases by ether inhalation on 4th day of nursing, and necropsied.

3.GROSS PATHOLOGY: 1) Parent animals

(1) General state observation
　Throughout the administration period and subsequent 14-day recovery period, animals were observed daily for life and death, appearance, and behavior.

(2) Detailed clinical observation
　The day before the start of administration and once a week thereafter, when the animal is removed from the cage and when it is removed from the cage in an aluminum open field (370 W x 560 D x 40 H mm) outside the cage Easiness, body tension (relaxation to tonicity), skin (color), fur, napping, eye secretions, closed eyelids, eyeball protrusion, lacrimation, nasal discharge (dirt), salivation, lower abdominal coat urine Contamination due to stool, stool around the anus, vocalization, breathing, posture, convulsions, tremor, exploratory behavior (wakefulness), vigilance, spontaneous movement (activity), walking (staggered), abnormal behavior (self-biting, backward facing We observed 29 items such as walking), regularity (excessive hair repair, repetitive turning, etc.), consciousness disorder (confused, catalepsy, coma), limb muscle tone, urination and defecation.

(3) Sensory reflex function test
　Males, males once in the nursing period, males and females in the satellite group, auditory response, visual response, tactile response, pinna reflex, pain response, pupillary reflex, Ipsilateral flexor reflexes, eyelid reflexes, and forward reflexes were examined.

(4) Landing leg width, grip strength and spontaneous momentum measurement
　Males are treated for 41 days, females are treated once during the nursing period, and males and females in the satellite group are on the last treatment day and recovery on the 13th day. The width of the landing leg is measured (measures the width of the footprint when dropped from a height of 30 cm) , Grip strength of forelimbs and hind limbs (rat / mouse grip strength measurement device, MK-380R / FR, Muromachi machine) and males for 30 and 60 minutes, females for 30 minutes (spontaneous momentum measurement device, SUPERMEX, Muromachi machine) , Measured the number of movements in 60 minutes between compartments in the measuring device). Spontaneous momentum of females in the satellite group was measured for 60 minutes.

(5) Body weight and food consumption measurement
　Body weight was measured once a week and on the day of sacrifice. The amount of food consumption was measured once a week for one day (24 hours).

(6) Sexual cycle inspection
　For females, vaginal plaque smears were prepared by Giemsa staining until mating was confirmed following the habituation and quarantine period, and the sexual cycle stage was determined by microscopic examination. The mean sexual cycle was defined as the average number of days in which the late estrus I, in which only keratinocytes were scattered or agglomerated, returned.

(7) Mating and observation of labor conditions
　On the afternoon of the 15th day of administration, females in the same group were placed in male cages (one-to-one) and allowed to live together for up to 14 days until mating was confirmed. Mating was confirmed every morning at a certain time, and the day when sperm was confirmed in the vaginal plug formation or vaginal plaque was defined as day 0 of pregnancy. The state of parturition was also observed at the same time, and the day when the end of parturition was confirmed for each abdomen was defined as day 0 of nursing. From the observation results of mating and delivery, the mating rate (%) [(number of mating animals / number of live animals) × 100], conception rate (%) [(number of conceived females / number of mating females) × 100] and The birth rate (%) [(number of live births / number of pregnant females) x 100] and the duration of pregnancy were calculated for the cases in which delivery was confirmed (days from the 0th day of pregnancy to the day on which delivery was confirmed). The female satellite group did not mate.

(8) Urinalysis
　For males, on the 40th day of administration and on the 8th day of recovery in the satellite group, the animals are placed in metabolic cages for about 3 hours. The resulting urine is used to observe the appearance, test paper method [Multistics, Bayer Sankyo ] Qualitative examination of pH, occult blood, protein, sugar, ketone body, bilirubin and urobilinogen, and examination of sediment (URI-CELL solution, stained with Cambridge Chemical Products Co., Ltd.). Furthermore, urine volume, specific gravity (refractometer, Elmer Optics) and sodium and potassium (electrolyte automatic analyzer, NAKL-132, Toa Denki Kogyo) were measured for urine obtained after 18-hour storage.

(9) Hematology test
　Blood samples were collected from the abdominal aorta under ether anesthesia the day after the administration and recovery period. The animals were weeded from 5 pm the day before blood collection and were given water only. The collected blood is divided into 3 parts, and a part of it is treated with EDTA-2K to prevent coagulation, and with a multi-item automatic blood cell counter (E-4000, Sysmex), the red blood cell count (electric resistance detection method), hemoglobin amount (lauryl) Sodium sulfate-hemoglobin method), hematocrit value (pulse detection method), mean red blood cell volume (MCV), mean red blood cell pigment content (MCH), mean red blood cell pigment concentration (MCHC, calculated above), white blood cell count and platelet count (above, Electrical resistance detection method), smear specimens were prepared, and the reticulocyte count (microscopically stained with Brilliant cresyl blue) and white blood cell percentage (microscopically stained with May-Giemsa) were measured. In some cases, plasma is separated by clotting inhibition treatment with 3.8% sodium citrate solution, and the prothrombin time (PT, Quick one-step method) and activation part are measured using an automatic blood coagulation analyzer (KC-10A, Amerung). Thromboplastin time (APTT, ellagic acid activation method) was measured.

(10) Blood biochemistry test
　Serum is separated from a portion of the collected blood and analyzed using a biochemical automatic analyzer [JCA-BM8, JEOL] with total protein (Biuret method), albumin (BCG method), A / G ratio (calculated value), glucose , Triglyceride, total cholesterol (above, enzymatic method), total bilirubin (diazo method), urea nitrogen (Urease-UV method), creatinine (Jaff), AST, ALT, g-GTP, ALP (above, JSCC method) , LDH (SFBC method), cholinesterase (BTC-DTNB method), calcium (OCPC method) and inorganic phosphorus (enzymatic method), and sodium, potassium and chlorine using an automatic electrolyte analyzer [NAKL-132, Toa Denpa Kogyo Co., Ltd.] (Ion electrode method) was measured.

(11) Pathological examination
　For male slaughtered animals, the day after the 42nd day of administration, in the case of females who were delivered and nurturing well, 5 days were nurtured, and those who were not mated were 24 days after the mating period (the day after the 52nd day). In cases where mating was established but no delivery was observed until 4 days after the scheduled delivery, and in the satellite group on the 14th day after recovery, the blood was sacrificed under ether anesthesia, and the body surface, opening The mucosa and internal organs were observed macroscopically. In addition, weigh (absolute weight) the liver, kidney, adrenal gland, thymus, spleen, brain, heart, pituitary gland, thyroid gland and seminal vesicle and all male testes and epididymis of each male and female. Based on the body weight, the ratio to body weight (relative weight) was calculated. For females, the number of ovaries of the ovaries and the number of uterus implantations were examined, and the implantation rate (%) [(implantation number / luteal number) × 100] was calculated.

　For all cases, brain, pituitary gland, thyroid, thymus, trachea / lung (fixed after infusion), stomach, intestine, heart, liver, spleen, kidney, adrenal gland, bladder, testis, epididymis, prostate, seminal vesicle , Ovary, uterus, spinal cord (neck, chest, lumbar region), sciatic nerve, bone marrow (femur), lymph node (cervical lymph node, mesenteric lymph node), mammary gland, and other grossly abnormal sites, The cells were fixed and preserved with 10% neutral phosphate buffered formalin solution (testis and epididymis were pre-fixed with Bouin's solution). Histopathological examinations were performed on these preserved organs in both sexes in the control and 250 mg / kg groups. All cases were examined for gross abnormalities in each group. In addition, for males and females who were not mated and females who were mated but were not delivered until 4 days after delivery, the testes, epididymis, prostate, and seminal vesicles were used in males, and the ovaries, uterus, and pituitary in females. Were inspected. In the examination, paraffin sections were prepared according to a conventional method, followed by HE staining and microscopic examination. The testis of the control group and the 1000 mg / kg group were subjected to PAS staining and spermatogenesis cycle tests (stages II, III, V, VII and XII).

2) Newborn items
(1) Number of babies, sex ratio and outer surface observation

　After confirming the completion of labor, the number of babies in each abdomen (the total number of babies born and dead) was examined. Gender was determined and sex ratio for each group was calculated. Newborns were observed for abnormalities in the outer surface including the oral cavity.　

(2) General state observation

　Check the general condition and life and death every day. I asked.

(3) Body weight measurement

　For newborns, the total body weight of each abdomen was measured for both sexes on days 0 and 4 of breastfeeding, and the average body weight per animal was calculated.

(4) Pathological examination

　In each case of death, the surviving cases were exsanguinated under ether anesthesia on the fourth day of nursing and the major organs in the thoracoabdominal region were visually observed.

4. Observations and examinations performed & frequency
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Monthly
- Cage side observations checked in table [No.?] were included. General condition and mortality

DETAILED CLINICAL OBSERVATIONS: No data
- Time schedule: No data

BODY WEIGHT: Yes
- Time schedule for examinations: Monthly

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes, Monthly
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data

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

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

OPHTHALMOSCOPIC EXAMINATION: No data
- Time schedule for examinations: No data
- Dose groups that were examined: No data

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

CLINICAL CHEMISTRY: No data
- Time schedule for collection of blood: No data
- Animals fasted: No data
- How many animals: No data
- Parameters checked in table [No.?] were examined. No data

URINALYSIS: No data
- Time schedule for collection of urine: No data
- Metabolism cages used for collection of urine: No data
- Animals fasted: No data
- Parameters checked in table [No.?] were examined. No data

NEUROBEHAVIOURAL EXAMINATION: No data
- Time schedule for examinations: No data
- Dose groups that were examined:v
- Battery of functions tested: sensory activity / grip strength / motor activity / other: No data

OTHER: No data

Sacrifice and pathology:

2.GROSS PATHOLOGY: Yes,
Males: After fasting period after the end of the administration on the final administration day, necropsy was done by exsanguinating with lethality under deep anesthesia with pentobarbital sodium at the next day (day corresponding to 43 days of administration). At that time, weights of heart, liver, kidney, thymus, testis and epididymis were measured for all cases. Of these organs, the testis and epididymis were fixed and stored in Bouin's solution, and other organs and brain, spleen, adrenal gland and bladder were fixed and fixed in 10% formalin.

Females: Females delivered were sacrificed on the fourth day of nursing, females that did not deliver but did not deliver to the 25th day of pregnancy, females who did not copulate were exsanguinated / lethal under pentobarbital sodium anesthesia at the end of the mating period and autopsied. Observation of organs / tissues with the naked eyes. At the same time, the organ weights were measured for the heart, liver, kidney and thymus, and the specific body weight (relative weight) was also calculated. Ovary and uterus were excised in both cases of pregnancy and infertility, the ovaries counted the number of pregnant lutein under a stereoscopic microscope, the number of implantation was counted for the uterus, and the implantation rate [(implantation number/pregnancy corpus luteum number) × 100] was calculated.

HISTOPATHOLOGY: Yes
Males: Organ examined: Brain, heart, liver, slpeen, thymus, kidney, urinary bladder, adrenal gland, ovary, testis and epididymis were examined. The testis and epididymis were fixed and stored in Bouin's solution, and other organs and brain, spleen, adrenal gland and bladder were fixed and fixed in 10% formalin. These organs were subjected to paraffin sections according to a conventional method for 1000 mg/Kg group and 0 mg/Kg group and subjected to histopathological examination by hematoxylin-eosin staining.

Females: The brain, heart, liver, spleen, kidney, bladder, adrenal gland, thymus, uterus and lesion were fixed in 10% formalin solution and ovaries were stored in Bouin's solution (10% formalin solution for long term storage). For all of the groups and control groups, histopathological examination of the above mentioned organs (except ovaries, uterus and lesions) was performed. However, histopathological examination was also performed on ovaries for females that had been mated but were infertile and females that did not copulate..

3. GROSS PATHOLOGY: YesMales were killed on day 43 and females were killed on day 5 of lactation. Recovery group males were sacrificed on day 57 and satellite group of the recovery females were killed on day 57.

HISTOPATHOLOGY: No data available

4. GROSS PATHOLOGY: Yes, Gross pathology was performed on animals that died.

HISTOPATHOLOGY: No data

Other examinations:

2.Reproductive performance of F0 animals, development and body weight of F1 pups up to day 4 of lactation and morphological findings of F1 pups were examined.
3.No data available

Statistics:

2.For mating rate and conception rate, chi 2 test including Yates' correction was conducted. Regarding histopathological findings, one sided tests were performed on Mann-Whitney U test for grade separated data and Fisher direct probability for total positive grade values. For the other data, the value obtained for each individual or average value for each litter was set as one sample, and the variance uniformity of each group was first tested by the Bartlett method. As a result, when the variance is made uniform, a oneway type of variance analysis is performed, and if significance is found between the groups, if the number of animals in each group is the same, the Dunnett method using, if not identical Scheff were assayed for differences in mean values between the control group and docosanoic each treatment group using the method. If the variance was not uniform or there was a group where the variance was 0, Kruskal-Wallis rank test was performed, and when significance was found between the groups, the control group and docosanoic acid Dunnett method or Scheff for the difference between the administration groups was carried out method test. The level of significance was set at 5% and 1%.
3. No data available
4. Average body weights without standard deviation were recorded for each group at approximately weekly intervals. There was no statistical analysis of the data.

Clinical signs:

no effects observed

Description (incidence and severity):

2.Males: No change in general condition was noted in any of the treatment groups.
Females: At 100 mg/Kg, from 42nd day of dosing, piloerection, blood like discharge from the vaginal opening in one females followed by coat contamination on 43-44 day administration.

3.No changes attributable to the administration of the test substance were observed during the treatment period and during the recovery period.

Mortality:

no mortality observed

Description (incidence):

2.Males/Females: No abnormalities in death were noted in any of the treatment groups.

3.One female in the 300 mg / kg group had a small mass in the lower abdomen after 40 days of administration. In other animals, there was no change in general status or death throughout the treatment and recovery period.

4.mortality observed, treatment-related
At the end of the study, 3, 2, 7 and 2 of 20 rats died from the control, 0.5, 1.0 and 3.0% groups, respectively

Body weight and weight changes:

no effects observed

Description (incidence and severity):

2.Males: The body weight gain increased from 8 to 15 days in the group administered 100 mg / kg significantly (p<0.01) compared with the control group, and the cumulative increase from the administration start date was also significantly higher Values after 29 days increased significantly (p <0.05) than the control group. However, in the group administered with 300 mg / kg or more, there was no significant difference between the control group and body weight and weight gain at any time.

Females: No significant difference in body weight and body weight gain between control group and docosanic acid administration group before breeding, pregnancy period and nursing period was noted.


4.no effects observed
Only minor effects on the weight development in the rats were observed

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

2.Males: No significant difference between the control group and docosanic acid administration group at any time of feeding intake was observed.

Females: No significant difference between the control group and docosanic acid administration group for food consumption before mating and pregnancy was observed. In the nursing stage, the consumption of 0 to 4 days of nursing during the 100 mg / kg administration group decreased significantly (p <0.01) compared with the control group, but in the group administered with 300 mg / kg or more, There was no significant difference between the control group.

3.There were no changes due to administration of the test substance during the administration period or recovery period.

4. no effects observed
There were no serious effects on the acceptability of the feed

Food efficiency:

not specified

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

not specified

Ophthalmological findings:

not specified

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

2.Males: The average red blood cell hemoglobin concentration (MCHC) decreased significantly (p <0.01) in comparison with the control group in the group administered with 300 mg / kg or more. No significant differences were found between the control group and docosanic acid administration group for other test items.

3.No change was observed after administration of the test substance. In the test at the end of the administration period, the activated partial thromboplastin time (APTT) of males in each test substance administration group was generally slightly lower than the control group, and the 100 mg / kg and 300 mg / kg groups There was a significant difference in the dose, but there was no dose-related relationship with the change.
In the examination at the end of the recovery period, there was no significant change in each examination item.

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

2.Males: Alkaline phosphatase activity decreased significantly (p <0.05) in the docosanic acid administration group and in the glucose concentration 1000 mg / kg administration group, respectively, as compared with the control group. In addition, the concentration of total protein and calcium decreased significantly (p <0.05, p <0.01) compared with the control group and the chlorine concentration increased significantly (p <0.05) in the group administered with 300 mg / kg. However this change was not dose related.

3.Examination at the end of the treatment period showed significant high levels of inorganic phosphorus in females in the 300 mg / kg and 1000 mg / kg groups. Although these values ​​were slightly higher than those in the control group and 100 mg / kg group, the dose correlation was not clear.
At the end of the recovery period, female cholinesterase in the 1000 mg / kg group showed a significantly high value.

Urinalysis findings:

no effects observed

Description (incidence and severity):

3.Male urine test:There were no significant changes in the examination during the administration period and recovery period.

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

2.Males: In the 100 mg / kg administration group, the actual weight and the specific body weight value of the liver increased significantly (p <0.05) compared to the control group, but was not considered to be dose related change. Regarding other organs, no significant difference was observed between the control group and docosanic acid administration group.

Females: The actual weight of the kidney decreased significantly (p <0.05) in the 100 mg/Kg dose animals compared with the control group, but this effect was not the dose dependent change. No significant difference was observed between the control group and docosanic acid administration group in other organs.

3.effects observed, treatment-related
At the end of the treatment period, males in the 100 mg / kg group had a significantly lower absolute seminal vesicle weight and females at 100 mg / kg had a significantly lower relative spleen weight. However, it was not a dose-related change.

At the end of the recovery period, the pituitary gland in males in the 1000 mg / kg group and the thyroid gland in females in the 1000 mg / kg group had significantly lower relative weights.

Gross pathological findings:

effects observed, non-treatment-related

Description (incidence and severity):

2.Males/ Females: No treatment related effects due to docosanoic acid administration were noted
3.
There were no changes due to the administration of the test substance. There was no change in males and females even in pairs where mating was not established or mating was established but did not become pregnant. The findings observed independently of test substance administration were as follows: at autopsy at the end of the administration period, the red area of ​​the thymus was in one male in the control group, and in the general condition observation in the 300 mg / kg group female. A tumor palpable in one of the animals was confirmed as a 2.0 x 2.5 cm large tumor under the right inguinal region. At autopsy at the end of the recovery period, enlargement of the spleen with a thickened capsule was observed in one male in the control group and one female in the 1000 mg / kg group in the red area of ​​the thymus.
4.no effects observed
No characteristic gross pathology was evident from the autopsies performed on the respective experimental groups.

Neuropathological findings:

no effects observed

Description (incidence and severity):

3.Sensory reflex function test :There was no significant change in each test item at 6 weeks after administration and 2 weeks after recovery.

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

2.Males: No significant differences were found between the control group and docosanic acid administration group for any histopathological findings.

The findings include: Myocardial degeneration of heart was observed in the animals of the control group and 1000 mg / kg administration group. Periportal lipidation of the liver was observed in the control group and 1000 mg / kg administration group, but there was no difference in frequency and degree between both groups. In addition, focal necrosis was found in one of the control group. Brown pigmentation and extramedullary hematopoiesis of spleen were observed in the control group and 1000 mg / kg administration group, but there was no difference in frequency and degree. Cortical basophilic tubules and eosinophilic bodies in kidneys were found in the control group and 1000 mg / kg group, but there was no difference in frequency and degree between the two groups. In addition, fibrosis was observed in kidneys in one of the control group. Fibrosis of the adrenal gland was observed in one of the 1000 mg / kg administration group, and there was no abnormality. Atrophy of seminiferous tubules was found in 2 animals in the 1000 mg / kg administration group, one on both sides and the other on one side, but both were localized and extremely mildly changed. There was no other abnormality. Sperm granulomas were found in the epididymis in one of the control group, and there was no abnormality. No abnormality was noted in brain, thymus and bladder.

Females: Deposition of minerals in the thalamus was noted in 1000 mg / kg administration group and no abnormality was noted. Focal necrosis was found in one of the control group and in all animals of 1000 mg/Kg group and also fibrosis was noted. Brown pigmentation and extramedullary hematopoiesis of the spleen were observed in the control group and 1000 mg / kg administration group, but there was no difference in frequency and degree between the two groups. Atrophy of the thymus and hemorrhage was observed in the control group and 1000 mg / kg administration group but no difference in control and treated animals was found. Cortical basophilic renal tubule and renal pelvic dilation of the kidneys were observed in the control group and 1000 mg/kg administration group. Necrosis was observed in one cortex in the control group, and there was no abnormality. No abnormality was noted in heart, bladder, ovaries of the unexpanded and infertile cases.

3.no effects observed
There were no changes due to the administration of the test substance. The testicular spermatogenesis cycle test showed no significant change.

Histopathological findings: neoplastic:

not specified

Description (incidence and severity):

3.effects observed, treatment-related
The findings that were considered to be spontaneous lesions unrelated to the administration of the test substance include cardiac myocardial degeneration / fibrosis, lung foam cell aggregation and arterial wall mineralization, hepatic steatosis of the liver and microgranulomas, Isolated kidney cyst, cortical lymphocyte infiltration, hyaline cast, mineralization and cortical fibrosis of the spinal cord boundary, and thymic hemorrhage are common to control group and 1000 mg / kg group, or only to control group A low incidence was observed. In addition, proximal tubular epithelial hyaline droplets in male kidneys and extramedullary hematopoiesis and brown pigmentation in male and female spleens were observed with high incidence, but the incidence and changes between 1000 mg / kg and control groups There was no difference in the degree of. In the 1000 mg / kg group only, mild prostatic stromal lymphocyte infiltration was observed in one male, and interstitial inflammation in the lung and focal necrosis in the liver were observed in one female. However, these changes were found spontaneously in rats and were judged to be independent of the administration of the test substance.

The subcutaneous mass observed at autopsy irrespective of the administration of the test substance was a fibroadenoma of the breast (benign). Hemorrhage was observed in the red area of ​​the thymus, and spleen with encapsulated fibrosis was observed in the enlarged spleen.

In one pair of pregnancy failure found in the control group, male testicular sexual epithelial degeneration, epididymal epididymal sperm reduction and prostate atrophy were observed in female pituitary, ovary and uterus. No change was observed. There was no change in male and female reproductive organs and female pituitary gland in a pair of mating failure in the 1000 mg / kg group.

Other effects:

not specified

Description (incidence and severity):

3.no effects observed
Landing leg width, grip strength and spontaneous momentum:
During the treatment period, no changes were observed.
Examination during the recovery period showed a significant increase in locomotor activity in the 60-minute start period of females in the 1000 mg / kg group. However, there was no significant difference in the amount of spontaneous exercise during the 30 minutes from the start of the measurement.

Details on results:

3. Through the administration and recovery periods, there were no changes attributable to administration of the test substance.

Dose descriptor:

NOAEL

Remarks:

2.

Effect level:

1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

body weight and weight gain

clinical biochemistry

clinical signs

food consumption and compound intake

gross pathology

haematology

histopathology: neoplastic

histopathology: non-neoplastic

mortality

neuropathology

organ weights and organ / body weight ratios

urinalysis

other: Sensory reflex function test;Landing leg width, grip strength and spontaneous momentum

Remarks on result:

other: No effect observed

Dose descriptor:

NOAEL

Remarks:

3.

Effect level:

1 000 other: mg/Kg/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No effects attributable to the test chemical were noted at the mentioned dose level

Dose descriptor:

NOAEL

Remarks:

4.

Effect level:

1 500 other: mg/Kg/day

Based on:

test mat.

Sex:

male

Basis for effect level:

other: No significant effects were noted at the mentioned dose level

Critical effects observed:

no

System:

other: not specified

2.

The conception rate also had no effect oftest chemical. There was no effect of test chemical administration on pregnancy period, maternal pregnancy rate, childbirth rate, labor condition and nursing condition, as well as survival, body weight, sex ratio and morphology of the infant.

Conclusions:

The No Observed Adverse Effect Level (NOAEL) for Octadecyl stearate is considered to be 1000 mg/Kg/day.

Executive summary:

Data available for the various test chemicals was reviewed to determine the toxic nature of the test chemical. The studies are as mentioned below:

 

A combined repeated dose toxicity and reproductive/developmental screening test (OECD TG 422) was performed to determine the toxicity likely to arise after repeated oral exposure to test chemical in male and female Sprague-Dawley rats.Groups of 13 rats/sex/dose were orally administered (gavage) 0, 100, 300 and 1000 mg of test chemical per kg of body weight. Rats in the control group were treated with corn oil (vehicle) under the same conditions as the test chemical administration group. In male rats, the administration period was two weeks prior to mating, 2 weeks of mating and 2 weeks after the completion of the mating period (42 days). In females, in addition to maximum four weeks pre-mating and mating period, they were exposed through pregnancy until day 3 of post-delivery (PDN 3). The repeated exposure to the test substance did not induce mortality or changes in general conditions of anyod thedosed animals. The mean body weight and food consumption of both males and females were comparable with controls. At day 43 males were necropsied, the heart, liver kidney and thymus were weighed then tissues were fixed, and histopathological examination were performed. From male animals, blood was also collected at necropsy for clinical biochemistry and haematology. The mean red blood cell haemoglobin concentration (MCHC) decreased significantly compared to the control group in the 300 and 1000 mg/kg dose groups, but no further significant difference was observed. Clinical biochemistry showed a significantly decreased alkaline phosphates activity in all test chemical-treated males and the glucose level dropped significantly in 1000 mg/kg group compared with controls.However, alterations of blood parameters were not accompanied by any changesfound in other examination including organ weights andhistopathological examination, therefore they were considered to be accidental changes. No treatment-related effects on the organ weights of heart, liver kidney and thymus were observed in treated males. Similarly, there were no obvious histopathological lesions in heart, liver kidney and thymus that could be attributed to administration of test chemical. Female animals (both mated and infertile) were also necropsied and organ weights were measured for heart, liver, kidney, and thymus. Organs were fixed and tissue samples were prepared for histopathological analysis. Autopsy, organ weight measurements and histopathological analysis did not show any alterations attributable to the administration of test chemical in any dose group of females.In summary,no treatment-related adverse effects were observed by haematology, organ weight measurements and histopathology in any dose group in both sexes, when male and female SD rats were administered 100, 300 and 1000 mg/kg bw/day test chemical for minimum 28 and maximum 42 days. Based on the observations of the present study,the No Observed Adverse Effect Level (NOAEL) for repeated dose toxicity by oral route was considered as 1000 mg/kg/day.

 

In another Combined Repeat Dose and Reproductive/Developmental Toxicity Screening Test was performed to determine the repeated dose oral toxic nature of the test chemical. 12 males and 12 females respectively were dosed with the test compound dissolved in corn oil at dose levels of 0, 100, 300 or 1000 mg/Kg/day. The duration of study was 42 days for males and from 14 days before mating to day 4 of lactation for females. Satellite group females and recovery group males were included in the study. Terminal killing was performed for males on day 43, females on day 5 of lactation, recovery group males on day 57 and satellite group for the recovery of females on day 57. No changes attributable to the administration of the test substance were observed during the treatment period and during the recovery period.One female in the 300 mg / kg group had a small mass in the lower abdomen after 40 days of administration. In other animals, there was no change in general status or death throughout the treatment and recovery period.During the recovery period, the weight gain of males in the 1000 mg / kg group showed a significantly high value, but this was rather due to the tendency of the weight gain of the control group to be low.There were no changes due to administration of the test substance during the administration period or recovery period.No change was observed after administration of the test substance. In the test at the end of the administration period, the activated partial thromboplastin time (APTT) of males in each test substance administration group was generally slightly lower than the control group, and the 100 mg / kg and 300 mg / kg groups There was a significant difference in the dose, but there was no dose-related relationship with the change. In the examination at the end of the recovery period, there was no significant change in each examination item.Examination at the end of the treatment period showed significant high levels of inorganic phosphorus in females in the 300 mg / kg and 1000 mg / kg groups. Although these values ​​were slightly higher than those in the control group and 100 mg / kg group, the dose correlation was not clear. At the end of the recovery period, female cholinesterase in the 1000 mg / kg group showed a significantly high value.At the end of the treatment period, males in the 100 mg / kg group had a significantly lower absolute seminal vesicle weight and females at 100 mg / kg had a significantly lower relative spleen weight. However, it was not a dose-related change.At the end of the recovery period, the pituitary gland in males in the 1000 mg / kg group and the thyroid gland in females in the 1000 mg / kg group had significantly lower relative weights.There were no changes due to the administration of the test substance. There was no change in males and females even in pairs where mating was not established or mating was established but did not become pregnant. The findings observed independently of test substance administration were as follows: at autopsy at the end of the administration period, the red area of ​​the thymus was in one male in the control group, and in the general condition observation in the 300 mg / kg group female. A tumor palpable in one of the animals was confirmed as a 2.0 x 2.5 cm large tumor under the right inguinal region. At autopsy at the end of the recovery period, enlargement of the spleen with a thickened capsule was observed in one male in the control group and one female in the 1000 mg / kg group in the red area of ​​the thymus.There was no significant change in each test item at 6 weeks after administration and 2 weeks after recovery.There were no changes due to the administration of the test substance. The testicular spermatogenesis cycle test showed no significant change.The findings that were considered to be spontaneous lesions unrelated to the administration of the test substance include cardiac myocardial degeneration / fibrosis, lung foam cell aggregation and arterial wall mineralization, hepatic steatosis of the liver and microgranulomas, Isolated kidney cyst, cortical lymphocyte infiltration, hyaline cast, mineralization and cortical fibrosis of the spinal cord boundary, and thymic hemorrhage are common to control group and 1000 mg / kg group, or only to control group A low incidence was observed. In addition, proximal tubular epithelial hyaline droplets in male kidneys and extramedullary hematopoiesis and brown pigmentation in male and female spleens were observed with high incidence, but the incidence and changes between 1000 mg / kg and control groups There was no difference in the degree of. In the 1000 mg / kg group only, mild prostatic stromal lymphocyte infiltration was observed in one male, and interstitial inflammation in the lung and focal necrosis in the liver were observed in one female. However, these changes were found spontaneously in rats and were judged to be independent of the administration of the test substance.The subcutaneous mass observed at autopsy irrespective of the administration of the test substance was a fibroadenoma of the breast (benign). Hemorrhage was observed in the red area of ​​the thymus, and spleen with encapsulated fibrosis was observed in the enlarged spleen.In one pair of pregnancy failure found in the control group, male testicular sexual epithelial degeneration, epididymal epididymal sperm reduction and prostate atrophy were observed in female pituitary, ovary and uterus. No change was observed. There was no change in male and female reproductive organs and female pituitary gland in a pair of mating failure in the 1000 mg / kg group.Examination during the recovery period showed a significant increase in locomotor activity in the 60-minute start period of females in the 1000 mg / kg group. However, there was no significant difference in the amount of spontaneous exercise during the 30 minutes from the start of the measurement. The No Observed Adverse Effect Level (NOAEL) for the test chemical is considered to be 1000 mg/Kg/day.

 

Furthermore, repeated dose oral toxicity study was performed to determine the toxic nature of the test chemical. The study was performed using male rats. The test chemical was mixed with feed and used at dose level of 0, 0.5, 1.0, or 3.0 % (0, 250, 500 or 1500 mg/Kg/day). During the study, the test animals were observed for body weights, feed consumption, mortalities and general appearance on a monthly basis. Gross pathology was performed on animals that died.Average body weights without standard deviation were recorded for each group at approximately weekly intervals. There was no statistical analysis of the data. There were no serious effects on the acceptability of the feed and had only minor effects on the weight development in the rats. At the end of the study, 3, 2, 7 and 2 of 20 rats died from the control, 0.5, 1.0 and 3.0% groups, respectively. No characteristic gross pathology was evident from the autopsies performed on the respective experimental groups. Based on the observations made, theNo observed adverse effect level (NOAEL) for the test chemical is considered to be 1500 mg/Kg/day when male rats were exposed with the test chemical for 2 years.

 

Based on the abave detailed studies available, the No Observed Adverse Effect Level (NOAEL) of the test chemical in the rat via oral route of exposure is considered to be 1000 mg/kg body weight in male and female animals.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Quality of whole database:

Data is Klimisch 2 and from authoritative database.

Repeated dose toxicity: inhalation - systemic effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: inhalation

Data waiving:

exposure considerations

Justification for data waiving:

a short-term toxicity study does not need to be conducted because exposure of humans via inhalation in production and/or use is not likely as based on the provided thorough and rigorous exposure assessment

Endpoint conclusion

Quality of whole database:

Waiver

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - systemic effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: dermal

Data waiving:

exposure considerations

Justification for data waiving:

a short-term toxicity study does not need to be conducted because exposure of humans via the dermal route in production and/or use is not likely as based on the provided thorough and rigorous exposure assessment

Endpoint conclusion

Quality of whole database:

Waiver

Repeated dose toxicity: dermal - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Repeated dose toxicity: Oral

Data available for the various test chemicals was reviewed to determine the toxic nature of the test chemical. The studies are as mentioned below:

 

A combined repeated dose toxicity and reproductive/developmental screening test (OECD TG 422) was performed to determine the toxicity likely to arise after repeated oral exposure to test chemical in male and female Sprague-Dawley rats.Groups of 13 rats/sex/dose were orally administered (gavage) 0, 100, 300 and 1000 mg of test chemical per kg of body weight. Rats in the control group were treated with corn oil (vehicle) under the same conditions as the test chemical administration group. In male rats, the administration period was two weeks prior to mating, 2 weeks of mating and 2 weeks after the completion of the mating period (42 days). In females, in addition to maximum four weeks pre-mating and mating period, they were exposed through pregnancy until day 3 of post-delivery (PDN 3). The repeated exposure to the test substance did not induce mortality or changes in general conditions of anyod thedosed animals. The mean body weight and food consumption of both males and females were comparable with controls. At day 43 males were necropsied, the heart, liver kidney and thymus were weighed then tissues were fixed, and histopathological examination were performed. From male animals, blood was also collected at necropsy for clinical biochemistry and haematology. The mean red blood cell haemoglobin concentration (MCHC) decreased significantly compared to the control group in the 300 and 1000 mg/kg dose groups, but no further significant difference was observed. Clinical biochemistry showed a significantly decreased alkaline phosphates activity in all test chemical-treated males and the glucose level dropped significantly in 1000 mg/kg group compared with controls.However, alterations of blood parameters were not accompanied by any changesfound in other examination including organ weights andhistopathological examination, therefore they were considered to be accidental changes. No treatment-related effects on the organ weights of heart, liver kidney and thymus were observed in treated males. Similarly, there were no obvious histopathological lesions in heart, liver kidney and thymus that could be attributed to administration of test chemical. Female animals (both mated and infertile) were also necropsied and organ weights were measured for heart, liver, kidney, and thymus. Organs were fixed and tissue samples were prepared for histopathological analysis. Autopsy, organ weight measurements and histopathological analysis did not show any alterations attributable to the administration of test chemical in any dose group of females.In summary,no treatment-related adverse effects were observed by haematology, organ weight measurements and histopathology in any dose group in both sexes, when male and female SD rats were administered 100, 300 and 1000 mg/kg bw/day test chemical for minimum 28 and maximum 42 days. Based on the observations of the present study,the No Observed Adverse Effect Level (NOAEL) for repeated dose toxicity by oral route was considered as 1000 mg/kg/day.

 

In another Combined Repeat Dose and Reproductive/Developmental Toxicity Screening Test was performed to determine the repeated dose oral toxic nature of the test chemical. 12 males and 12 females respectively were dosed with the test compound dissolved in corn oil at dose levels of 0, 100, 300 or 1000 mg/Kg/day. The duration of study was 42 days for males and from 14 days before mating to day 4 of lactation for females. Satellite group females and recovery group males were included in the study. Terminal killing was performed for males on day 43, females on day 5 of lactation, recovery group males on day 57 and satellite group for the recovery of females on day 57. No changes attributable to the administration of the test substance were observed during the treatment period and during the recovery period.One female in the 300 mg / kg group had a small mass in the lower abdomen after 40 days of administration. In other animals, there was no change in general status or death throughout the treatment and recovery period.During the recovery period, the weight gain of males in the 1000 mg / kg group showed a significantly high value, but this was rather due to the tendency of the weight gain of the control group to be low.There were no changes due to administration of the test substance during the administration period or recovery period.No change was observed after administration of the test substance. In the test at the end of the administration period, the activated partial thromboplastin time (APTT) of males in each test substance administration group was generally slightly lower than the control group, and the 100 mg / kg and 300 mg / kg groups There was a significant difference in the dose, but there was no dose-related relationship with the change. In the examination at the end of the recovery period, there was no significant change in each examination item.Examination at the end of the treatment period showed significant high levels of inorganic phosphorus in females in the 300 mg / kg and 1000 mg / kg groups. Although these values ​​were slightly higher than those in the control group and 100 mg / kg group, the dose correlation was not clear. At the end of the recovery period, female cholinesterase in the 1000 mg / kg group showed a significantly high value.At the end of the treatment period, males in the 100 mg / kg group had a significantly lower absolute seminal vesicle weight and females at 100 mg / kg had a significantly lower relative spleen weight. However, it was not a dose-related change.At the end of the recovery period, the pituitary gland in males in the 1000 mg / kg group and the thyroid gland in females in the 1000 mg / kg group had significantly lower relative weights.There were no changes due to the administration of the test substance. There was no change in males and females even in pairs where mating was not established or mating was established but did not become pregnant. The findings observed independently of test substance administration were as follows: at autopsy at the end of the administration period, the red area of ​​the thymus was in one male in the control group, and in the general condition observation in the 300 mg / kg group female. A tumor palpable in one of the animals was confirmed as a 2.0 x 2.5 cm large tumor under the right inguinal region. At autopsy at the end of the recovery period, enlargement of the spleen with a thickened capsule was observed in one male in the control group and one female in the 1000 mg / kg group in the red area of ​​the thymus.There was no significant change in each test item at 6 weeks after administration and 2 weeks after recovery.There were no changes due to the administration of the test substance. The testicular spermatogenesis cycle test showed no significant change.The findings that were considered to be spontaneous lesions unrelated to the administration of the test substance include cardiac myocardial degeneration / fibrosis, lung foam cell aggregation and arterial wall mineralization, hepatic steatosis of the liver and microgranulomas, Isolated kidney cyst, cortical lymphocyte infiltration, hyaline cast, mineralization and cortical fibrosis of the spinal cord boundary, and thymic hemorrhage are common to control group and 1000 mg / kg group, or only to control group A low incidence was observed. In addition, proximal tubular epithelial hyaline droplets in male kidneys and extramedullary hematopoiesis and brown pigmentation in male and female spleens were observed with high incidence, but the incidence and changes between 1000 mg / kg and control groups There was no difference in the degree of. In the 1000 mg / kg group only, mild prostatic stromal lymphocyte infiltration was observed in one male, and interstitial inflammation in the lung and focal necrosis in the liver were observed in one female. However, these changes were found spontaneously in rats and were judged to be independent of the administration of the test substance.The subcutaneous mass observed at autopsy irrespective of the administration of the test substance was a fibroadenoma of the breast (benign). Hemorrhage was observed in the red area of ​​the thymus, and spleen with encapsulated fibrosis was observed in the enlarged spleen.In one pair of pregnancy failure found in the control group, male testicular sexual epithelial degeneration, epididymal epididymal sperm reduction and prostate atrophy were observed in female pituitary, ovary and uterus. No change was observed. There was no change in male and female reproductive organs and female pituitary gland in a pair of mating failure in the 1000 mg / kg group.Examination during the recovery period showed a significant increase in locomotor activity in the 60-minute start period of females in the 1000 mg / kg group. However, there was no significant difference in the amount of spontaneous exercise during the 30 minutes from the start of the measurement. The No Observed Adverse Effect Level (NOAEL) for the test chemical is considered to be 1000 mg/Kg/day.

 

Furthermore, repeated dose oral toxicity study was performed to determine the toxic nature of the test chemical. The study was performed using male rats. The test chemical was mixed with feed and used at dose level of 0, 0.5, 1.0, or 3.0 % (0, 250, 500 or 1500 mg/Kg/day). During the study, the test animals were observed for body weights, feed consumption, mortalities and general appearance on a monthly basis. Gross pathology was performed on animals that died.Average body weights without standard deviation were recorded for each group at approximately weekly intervals. There was no statistical analysis of the data. There were no serious effects on the acceptability of the feed and had only minor effects on the weight development in the rats. At the end of the study, 3, 2, 7 and 2 of 20 rats died from the control, 0.5, 1.0 and 3.0% groups, respectively. No characteristic gross pathology was evident from the autopsies performed on the respective experimental groups. Based on the observations made, theNo observed adverse effect level (NOAEL) for the test chemical is considered to be 1500 mg/Kg/day when male rats were exposed with the test chemical for 2 years.

 

Based on the abave detailed studies available, the No Observed Adverse Effect Level (NOAEL) of the test chemical in the rat via oral route of exposure is considered to be 1000 mg/kg body weight in male and female animals.

Repeated dose toxicity: Inhalation

Test chemical has very low vapor pressure of 2.16E-09 Pa. (1.62013e-11 mm Hg), so the potential for the generation of inhalable vapours is very low, also the normal conditions of use of this substance will not result in aerosols, particles or droplets of an inhalable size, so exposure to humans via the inhalatory route will be highly unlikely and therefore this end point for repeated dose toxicity by inhalation route of exposure was considered for waiver.

 

Repeated dose toxicity: Dermal

The acute dermal toxicity value for test chemical (as provided in section 7.2.3) is >2000 mg/kg body weight. Given the use of the chemical; repeated exposure by the dermal route is unlikely since the use of gloves is common practice in industries. Thus, it is expected that test chemical shall not exhibit 28 day repeated dose toxicity by the dermal route. In addition, there is no data available that suggests that test chemical shall exhibit repeated dose toxicity by the dermal route. Hence this end point was considered for waiver.

Justification for classification or non-classification

Considering the above discussion and applying the weight of evidence approach with no toxic effects noted, the test chemical is not likely to be toxic as per the criteria mentioned in CLP regulation.