Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Link to relevant study records
Reference
Endpoint:
screening for reproductive / developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was performed according to OECD guideline study with GLP compliance.
Qualifier:
according to
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
Eight-week-old Cr j: CD (SD) IGS rat (Nihon Challis • Riva Co., Ltd.) was purchased for 55 males and females and quarantined acclimatization for 13 days. During this period, observation of the general condition and measurement of body weight were carried out, and necropsy was performed for each of 3 males and females, after confirming that there was no abnormality, selecting animals that showed good growth and at 10 weeks of age It was used for the test. Body weight at the start of administration was 347.7~432 .2 g for male and 220.3~255.2 g for female.
Animals were exposed to a temperature of 24 °C +/- 2 °C (tolerance 21~27 °C), humidity 55 ± 10% (tolerance 35~75%), to 12 hours illumination (7:00 am at 7:00 am) and ventilation times 13~15 times / In the breeding room (No. 92) in the system C area, 23 stainless steel kegs (W 260 x H 200 x D 380 mm), 2~3 per sex during the quarantine conditioning period, 1 per sex during the administration period (One for each sex during the mating period), a polycarbonate keg with a bedding (Whiteflex, manufactured by Nihon Chara Riba Co., Ltd.) during the pregnancy and repatriation period (W 6 5 × H 1 85 × D 4 25 mm), one for each including fine-grown children during the question) housed and raised. Incidentally, the actual measured value of the temperature is a maximum of 26 ° C., the lowest 21 °C, the measured humidity was the maximum 57%, the lowest 50%. Well water (about 2 ppm) to which solid feed (MF, manufactured by Oriental Yeast Industry Co., Ltd.) was added as drinking water and sodium hypochlorite was added freely by an automatic water supply device or a water supply bottle, respectively. For feed and bedding, we analyzed at the Japan Food Analysis Center, Japan Foundation Analysis Center and drinking water was analyzed at Tsuruga Seinan Knu Research Center Co., Ltd., and confirmed that they conformed to acceptance criteria. Breeding equipment and feeds must be sterilized by high pressure steaming, the keg mounts and polycarbonate top cover are once every 4 weeks, the stainless steel key paper once a week, made of polycarbonate Jewel
The water bottle is exchanged at least once a week, the stainless steel keeper dish is exchanged three times a week (however, during the mating period every day), the breeding room is cleaned every day, the disinfectant is soaked in a mop I wiped it.

Route of administration:
oral: gavage
Vehicle:
CMC (carboxymethyl cellulose)
Details on exposure:
Routes of administration are specified in the OECD Test Method Guidelines and bioassociated with oral administration which is one of the anticipated route of exposure to human beings is 14 days before mating and after 35 days including the mating period. For a total of 49 days, females were administered for 14 days before mating, for a mating period (maximum 14 days), pregnancy period and up to 3 days of consultation, once daily, using gavage. The dose volume was 2.5 ml / kg, and the dosing volume for each animal was calculated based on the latest weights for male and females before mating and mating period, and for females after mating, the body weight at 0 day of pregnancy, respectively. A 0.5 W / V% CMC-Na solution was similarly administered to the control group.
Details on mating procedure:
Mating is done at around 4 pm on 15th day of administration, by a method of making them live together for one night in both sexes (12 weeks old). In the next morning, the mating was confirmed by the presence of sperm or plaster in land plants, and that day was designated as O pregnancy day. In addition, crossbreeding was conducted within the same group, and the mating period was a maximum of 2 weeks.
Analytical verification of doses or concentrations:
no
Duration of treatment / exposure:
male; 49 days: female; for 14 days before mating to day 3 of lactation
Frequency of treatment:
one administration/day
Details on study schedule:
Premating exposure period :
Male : 14 day
Female : 14 day
Duration of test: male: 49 day
female: 39-53 day
Remarks:
Doses / Concentrations:
0, 4, 20, 100 mg/kg bw/day
Basis:

No. of animals per sex per dose:
12
Control animals:
yes, concurrent vehicle
Parental animals: Observations and examinations:
General condition observation and body weight and food consumption measurement
For both males and females, observation of general condition and confirmation of viability and death for all cases were conducted before and after daily administration I went twice.
For body weight, in male, measurement was made twice a week throughout the administration period. In females, twice a week during the administration period and the mating period before mating, 0, 4, 7, 10, 14, 17 and 21 days during pregnancy during pregnancy, during the clearing period, it was measured at a clearance 0 (day of delivery) and on the 4th.
Regarding food intake, in males, measurements were made twice a week during the administration period excluding the mating period. In females, twice a week during the administration period before mating, 1, 4, 7, 10, 14, 17 and 21 days pregnancy during pregnancy, and on day 1 and day 4 of clearance during daily feeding .
Oestrous cyclicity (parental animals):
For females, scatterers are collected at regular time in the morning every day for 15 days from the administration start date (1 day of administration) and conducted a sexual cycle examination. In the sexual cycle examination, the number of estrus times was calculated, and the number of days of hearing from the estrus period until the next estrus period was taken as the estrus cycle period days and the average estrus cycle was calculated.
Litter observations:
For childbirth, at the time of childbirth, the number of births, the number of newborn babies, the number of stillborn babies, the sex of newborn baby and the abnormal outer table were inspected. For newborn babies, weight is measured for each individual on the day of birth and on day 4 of fertility and the birth rate [(number of newborn infants / implantation number) × 100], the stillbirth rate [(the number of stillborn infants / the number of births) × 100] and the survival rate on 4th day of newborn baby [(number of births on 4th day of clearance / number of newborns) × 100] were calculated.
Postmortem examinations (parental animals):
Mortality in the course of the test was autopsied promptly, organ weight measurement and histopathological examination were performed. Organ weight was used as reference data. Body weight, food intake, hematology examination value, blood biochemical examination value, number of days required for mating, number of sexual cycle examination values (number of matings, period of estrus), organ weight, pregnancy period, number of corpus luteums, number of implantation traces, total birth For the number of babies, the number of newborn babies and the weight of newborn babies, the mean value and standard deviation were determined for each group, and the homogeneity of dispersion was first tested by the B artlett method between the control group and the test substance administration group. When the variance is uniform, comparison with the control group is carried out by using the multiple comparison test of Du nnt t t. If the dispersion is not uniform,
Comparison with the control group was done using multiple comparison test of S t ee l. With regard to mating rate, receiving (a) gestation rate, birth rate and neonatal sex ratio, by χ 2 test, the mortality rate, the birth rate and the 4th day of newborn baby
For the survival rate of WI l o o x o n for the pathological histology exam.
Postmortem examinations (offspring):
Eyebrows On the 4 th day all the newborn infants underwent esthetic anesthesia and macroscopic observation of the organs and tissues was carried out. For postnatal infants (including stillborn infants and deceased children), they were fixedly stored in pure ethanol on a unit basis after necropsy.
Statistics:
The significance level was set to 5% in both cases where comparison was made between the control group and each treatment group list by Mann-Whitney U test. The measured values for newborn babies were processed in units of stomach.
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
In males, in the group of 20 mg / kg or less, there was no death and no change was observed in the general condition.
In females, two subjects in the 20 mg / kg group and 7 subjects in the 100 mg / kg group were autopsyed on the way because all the children died by 3 rd day nursing.
Dermal irritation (if dermal study):
not examined
Mortality:
mortality observed, non-treatment-related
Description (incidence):
In males, one patient in the 100 mg / kg group had decreased spontaneous locomotor activity and slow respiration before administration on 18th administration, and died before administration on 21th administration.
In females, one patient in the 100 mg / kg group died before administration on the 22nd day of pregnancy. In addition, in another one of the 100 mg / kg group, subcutaneous carcinoma of the chest was observed from 9th pregnancy to the autopsy day (4th day of clearance). No deaths occurred in the group of 20 mg / kg or less, no change was observed in the general condition.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
In male, suppression of body weight increase or tendency was observed throughout the administration period in the group of 20 mg / kg or more, and the total body weight gain during the administration period showed a significant low value.
In females, increase inhibition was observed on days 4 and 8 during the administration period before the mating in the 100 mg / kg group. In addition, during pregnancy and rest period, suppression of body weight gain or tendency was observed in the group of 20 mg / kg or more.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
In males, there was a decrease in the early stage of administration in the group of 20 mg / kg or more and then recovered, but again decreased in the latter stage of administration after the end of the mating period.
In females, there was a decrease in the early stage of administration in the group of 20 mg / kg or more, and then recovered, but eyelid growth.
It decreased again on the day. Incidentally, in the 100 mg / kg group, an increase was observed at the administration day 8 15 days.
In addition, as an accidental change without dose correlation, increase in pregnancy 21 days was observed in the 4 mg / kg group.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
Red blood cell count and hematocrit decreased and MCH and MCHC increased in the 100 mg / kg group. In the 100 mg / kg group, the number of reticulocytes also increased. Increase in MCH was also observed in the 20 mg / kg group.
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
Increases in albumin, A / G ratio, GOT, GPT, total cholesterol, phospholipids and total bilirubin and decreases in triglyceride, sodium and potassium were observed in the 100 mg / kg group. In addition, reduction of GPT, alkaline phosphatase and creatinine was observed in the 20 mg / kg group, but both were within the physiological variation range.
Urinalysis findings:
not examined
Behaviour (functional findings):
no effects observed
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
In males, the absolute and relative weight of the thymus decreased or decreased in the 100 mg / kg group, and the absolute and relative weights of the liver and accessory kidney increased or increased. In addition, in males, the absolute weight loss of the tiles was reduced in the 100 mg / kg group, and in the 20 mg / kg group, the relative weights of the lungs, kidneys, adrenal glands, testis and epididymis were increased, 100 mg / kg Increase in relative weight of brain, heart, lung, kidney, testis and epididymis in the group.
In females, an increase in the relative weight of the brain was observed in the 100 mg / kg group.
In addition, as an incidental change without dose correlation, the absolute weight increase of the adrenal gland was observed in males in the 4 mg / kg group, and in the females in the 20 mg / kg group, the absolute and relative weights of the heart were observed .
Gross pathological findings:
not specified
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Hypertrophy of central leukocytes was observed in 5 males and 7 females in the 100 mg / kg group. In males, testicular atrophy was seen in epididymal spermatogranuloma and 2 cases in control group and 3 cases in control group, 4 and 100 mg / kg group, respectively, and testicular atrophy was observed In one of the cases, reduction of sperm in the epididymal cavity was also observed. In females, one case of 100 mg / kg group showed proliferation of transitional epithelium of kidney and papilla, in which basophilic renal tubular epithelium basification and adenocarcinoma of mammary gland were each observed in each case It was done. In females, atrophy of the thymus was observed in 1, 3 and 1 cases of control group, 20 and 100 mg / kg group, respectively.
In the case of death, one males in the 100 mg / kg group treated with neutrophilic cell infiltration and granulation, neutrophilic cell infiltration and granulation, glandular erosion, edema of the lungs, atrophy of the tiles, Gland atrophy and out Blood, female erosion of the gland stomach, edema of the lung, atrophy of the tile and necrosis of the adrenal gland were observed.
Histopathological findings: neoplastic:
not specified
Reproductive function: oestrous cycle:
no effects observed
Description (incidence and severity):
In the sexual cycle examination, there was a difference in listening with the control group in the estrus count and estrus cycle in each group.
Reproductive function: sperm measures:
not examined
Reproductive performance:
no effects observed
Description (incidence and severity):
In the reproductive ability test, matings were observed in all cases in each group, but one in each of the control group and the 100 mg / kg group was infertile. Therefore, the mating rate was 100% in each group, the received (a) gestational rate was 91.6 7, 100, 100 and 9 1. 67% in the control group, 4 '20 and 100 mg / kg group, respectively, There was no difference between the control group and each group. Also in the number of days required for copulation, there was no difference in the difference between the control group and each group.
Key result
Dose descriptor:
NOAEL
Effect level:
100 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male
Basis for effect level:
clinical signs
body weight and weight gain
Key result
Dose descriptor:
NOAEL
Effect level:
4 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
female
Basis for effect level:
clinical signs
body weight and weight gain
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
There were no differences in the pregnancy period, the number of corpus luteums, the number of implantation traces, the number of births, the number of newborn babies, the birth rate and neonatal sex ratio in each group compared with the control group. No abnormality was found in each group in the neonatal exterior examination as well.
In the examination of the soybean season, in 2 cases in the 20 mg / kg group and 7 cases in the 100 mg / kg group, poor behavior such as regrowth, lactation, warming and the like of children was observed, and in these mother animals. All the children died, and in the group of 20 mg / kg or more, the survival rate of the newborn was decreased 4 days. Furthermore, in the 100 mg / kg group, both sexes showed a low value on the 4th day of the newborn's shooting.
Dermal irritation (if dermal study):
not examined
Mortality / viability:
mortality observed, non-treatment-related
Description (incidence and severity):
Furthermore, in the 100 mg / kg group, the increase in the mortality rate, the associated decrease in the fertility rate, and the decrease in female and male neonatal weight were observed. In addition, there was an increase in stillbirth rate in the 4 mg / kg group. However, similar changes were not observed in the 20 mg / kg group, so it was considered to be an accidental change.
Body weight and weight changes:
not examined
Food consumption and compound intake (if feeding study):
not examined
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
not examined
Histopathological findings:
not examined
Key result
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
20 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
not specified
Basis for effect level:
body weight and weight gain
Key result
Reproductive effects observed:
no

The NOAEL of 4 mg/kg bw/day for breast feeding females cannot be assigned to a reproductive toxicity unambigeously.

The lack of nursing activity can be due to the general stress caused by the toxicity of the substance or to specific effects of Benzoguanamine on the lactation.

Treatment related changes were only revealed after delivery of F0dams due to poor collection, nursing and heating for newborns at 20 mg/kg/day and more group.

STATISTICAL RESULTS: As for reproductive performance, no effects related to this substance were observed for the estrous cycle, numbers of corpora lutea and implantation, copulation index, conception index, and duration of mating. On examination after delivery, poor collection and heating of new borns were observed with dams of the 100 mg/kg group. Furthermore, the birth index decreased with increase of stillborns at this dose. No effects related to this substance were observed in terms of gestational days, number of litters and live newborns, gestation index and sex ratio. There were no external anomalies of pups. Examination during the lactation period, revealed poor collection, nursing and heating for newborns. A decrease in the viability index on day 4 of lactation was observed for dams of the 20 mg/kg or more group.

Findings of delivery of Fo dams treated orally with this subsutance

     Dose (mg/kg)

0

4

20

100

 

No. of copulated (male, female)

No. of impregnated (female)

12

11

12

12

12

12

12

11

 

    No. of dams

  11

12

 12

 10

 

Gestation index

100.00

100.00

100.00

90.90

 

   No. of poor nursing dams

 

 

 

 

 

           on day 0

 0

 0

 0

 21)

 

                day 4

 0

 0

  22)

  72)

 

     Mean no. ofnewborns/litter

15.45

14.45

13.75

15.20

 

            S.D. of number

3.14

1.66

3.70

2.25

 

   No. of stillborns

 0

  4*

 4

   43**

 

   No. of live newborns

170

173

161

109

 

   Mean no. of live newborns/litter

15.45

14.42

13.43

10.90

 

            S.D. of number

 3.14

 1.62

 3.45

 5.07

 

    Birth index

 100.00

 97.74

 97.58

71.71**

 

    Sex ratio of live newborns

   0.87

 0.90

0.96

0.98

 

  Body weight of live newborns (g)

 

 

 

 

 

     male on day 0

 6.3

  6.8

  6.2

 5.1**

 

                    day 4

 9.6

 10.6

  9.9

 6.8**

 

     female on day 0

 6.0

   6.5

   5.8

 4.7**

 

                      day 4

 9.2

 10.1

   9.3

 5.8**

 

     Viability index

 99.41

 99.42

 75.78*

11.93**

 

  No. of external anomalies

0

0

0

 0

 

 

Gestation index = (Number of dams with live newborns/Number of pregnant females) x 100

Birth index = (Number of live newborns/Number of (stillborns + live newborns)) x 100

Viability index = [number of live newborns on day 4 after birth/number of live newborns] x 100

*: p < 0.05, **: p < 0.01 (significantly different from control)

1): All newborns were dead and counted as stillborn

2): All newborns were dead before day 4

 

Conclusions:
The parental NOAEL of reproductive toxicity was considered to be 100 mg/kg/day for males, 20 mg/kg/day for females and 4 mg/kg bw/day for breast feeding females based on the lack of nursing activity.
Executive summary:

In the OECD combined repeat dose and reproductive/ developmental (one generation)toxicity screening test [OECD TG 422], this substance was given for 49 days from 14 days before mating in males and from 14 days before mating to day 3 of lactation in females. At the 100 mg/kg bw group, one female died in gestation and another female was not impregnated. Birth index was decreased with increase in stillborns at the 100 mg/kg bw. All pups of two dams at the 20 mg/kg bw and seven dams at 100 mg/kg bw died due to the lack of nursing activity, and the viability index on day 4 after birth was consequently decreased in these groups. The body weights of pups were also decreased at birth and day 4 of lactation in the 100 mg/kg bw group. The decrease of litter size observed at the 100 mg/kg bw seems to be the chemical-induced effect although it is not statistically significant. No malformations or variations were observed in the pups.

In this study effects on birth index and body weight gain of newborns as reported. The NOAEL for these effects was set with 20mg/kg. The NOAEL for male rats was 10 mg/kg (highest dose applied). Female rats had a lower nursing activity for the group dosed 20 mg/kg bw and higher, the NOAEL for these effects was set with 4 mg/kg bw. The NOAEL for effects on F1 generation is closed to NOAEL from the 90 day study (19 mg/kg bw). The NOAEL for effects on nursing female rats (4 mg/kg bw) is lower than the NOAEL from the 90 day study (19 mg/kg bw). These results indicate that there might be a relevant potency for reproductive toxicity. It is not possible to finally evaluate, wether the effects on reproduction are the result of a generalized toxicity or wether the substance has a specific toxicity with respect to the endpoint.

The parental NOAEL of reproductive toxicity was considered to be 100 mg/kg/day for males, 20 mg/kg/day for females and 4 mg/kg bw/day for breast feeding females based on the lack of nursing activity.

Based on exposure considerations, the lower NOAEL (4 mg/kg bw/day) refering to the lack of breast feeding may not be taken into account for the assessment:

The substance is a monomer used for polymers and resins. RMM to minimise the exposure for the relevant subpopulation are taken.

Therefor the NOAEL (20 mg/kg bw/day) refering to the non-breast-feeding population is used for the risk characterisation.

Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

Benzoguanamine was administered to each 12 males and 12 females of Sprague-Dawley (Crj: CD) rats by gavage at doses of 0, 4, 20 and 100 mg/kg bw (vehicle; 0.5% CMC-Na solution) from 14 days before mating, until14 days after mating in males, and from 14 days before mating until day 3 of lactation in females (Anonymous 1997).

As for reproductive performance, no effects related to this substance were observed for the count of estrus, estrous cycle, numbers of corpra lutea and implantation, copulation index, conception index, duration of mating.

No effects ( number of litters, gestation index) were observed in dams in the 100 mg/kg bw/day group, with respect to delivery of F0.

The parental NOAEL of reproductive toxicity was considered to be 100 mg/kg bw/day for males, 20 mg/kg bw/day for females and 4 mg/kg bw/day for breast feeding females based on the lack of nursing activity.

The prenatal Developmental study shows that maternal toxicity determines fetal toxicity and therefore reprotoxic effects can be considered as secondary effects due to the general toxicity of the substance.

 


Short description of key information:
The parental NOAEL of reproductive toxicity was considered to be 100 mg/kg bw/day for males, 20 mg/kg bw/day for females and 4 mg/kg bw/day for breast feeding females based on the lack of nursing activity.

Justification for selection of Effect on fertility via oral route:
see Discussion

Effects on developmental toxicity

Description of key information
The NOAEL of developmental toxicity was considered to be 20 mg/kg bw/day based on decreases of birth index and body weight pups.
Link to relevant study records
Reference
Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was performed according to OECD guideline study with GLP compliance.
Qualifier:
according to
Guideline:
other: OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Details on test animals and environmental conditions:
Eight-week-old Cr j: CD (SD) IGS rat (Nihon Challis • Riva Co., Ltd.) was purchased for 55 males and females and quarantined acclimatization for 13 days. During this period, observation of the general condition and measurement of body weight were carried out, and necropsy was performed for each of 3 males and females, after confirming that there was no abnormality, selecting animals that showed good growth and at 10 weeks of age It was used for the test. Body weight at the start of administration was 347.7~432 .2 g for male and 220.3~255.2 g for female.
Animals were exposed to a temperature of 24 °C +/- 2 °C (tolerance 21~27 °C), humidity 55 ± 10% (tolerance 35~75%), to 12 hours illumination (7:00 am at 7:00 am) and ventilation times 13~15 times / In the breeding room (No. 92) in the system C area, 23 stainless steel kegs (W 260 x H 200 x D 380 mm), 2~3 per sex during the quarantine conditioning period, 1 per sex during the administration period (One for each sex during the mating period), a polycarbonate keg with a bedding (Whiteflex, manufactured by Nihon Chara Riba Co., Ltd.) during the pregnancy and repatriation period (W 6 5 × H 1 85 × D 4 25 mm), one for each including fine-grown children during the question) housed and raised. Incidentally, the actual measured value of the temperature is a maximum of 26 ° C., the lowest 21 °C, the measured humidity was the maximum 57%, the lowest 50%. Well water (about 2 ppm) to which solid feed (MF, manufactured by Oriental Yeast Industry Co., Ltd.) was added as drinking water and sodium hypochlorite was added freely by an automatic water supply device or a water supply bottle, respectively. For feed and bedding, we analyzed at the Japan Food Analysis Center, Japan Foundation Analysis Center and drinking water was analyzed at Tsuruga Seinan Knu Research Center Co., Ltd., and confirmed that they conformed to acceptance criteria. Breeding equipment and feeds must be sterilized by high pressure steaming, the keg mounts and polycarbonate top cover are once every 4 weeks, the stainless steel key paper once a week, made of polycarbonate Jewel
The water bottle is exchanged at least once a week, the stainless steel keeper dish is exchanged three times a week (however, during the mating period every day), the breeding room is cleaned every day, the disinfectant is soaked in a mop I wiped it.

Route of administration:
oral: gavage
Vehicle:
CMC (carboxymethyl cellulose)
Details on exposure:
Routes of administration are specified in the OECD Test Method Guidelines and bioassociated with oral administration which is one of the anticipated route of exposure to human beings is 14 days before mating and after 35 days including the mating period. For a total of 49 days, females were administered for 14 days before mating, for a mating period (maximum 14 days), pregnancy period and up to 3 days of consultation, once daily, using gavage. The dose volume was 2.5 ml / kg, and the dosing volume for each animal was calculated based on the latest weights for male and females before mating and mating period, and for females after mating, the body weight at 0 day of pregnancy, respectively. A 0.5 W / V% CMC-Na solution was similarly administered to the control group.
Analytical verification of doses or concentrations:
no
Details on mating procedure:
Mating is done at around 4 pm on 15th day of administration, by a method of making them live together for one night in both sexes (1 2 weeks old). In the next morning, the mating was confirmed by the presence of sperm or plaster in land plants, and that day was designated as O pregnancy day. In addition, crossbreeding was conducted within the same group, and the mating period was a maximum of 2 weeks.
Duration of treatment / exposure:
male; 49 days: female; for 14 days before mating to day 3 of lactation
Frequency of treatment:
one administration/day
No. of animals per sex per dose:
12
Control animals:
yes, concurrent vehicle
Maternal examinations:
General condition observation and body weight and food consumption measurement
For both males and females, observation of general condition and confirmation of viability and death for all cases were conducted before and after daily administration I went twice.
For body weight, in male, measurement was made twice a week throughout the administration period. In females, twice a week during the administration period and the mating period before mating, 0, 4, 7, 10, 14, 17 and 21 days during pregnancy during pregnancy, during the clearing period, it was measured at a clearance 0 (day of delivery) and on the 4th.
Regarding food intake, in males, measurements were made twice a week during the administration period excluding the mating period. In females, twice a week during the administration period before mating, 1, 4, 7, 10, 14, 17 and 21 days pregnancy during pregnancy, and on day 1 and day 4 of clearance during daily feeding .
Fetal examinations:
For childbirth, at the time of childbirth, the number of births, the number of newborn babies, the number of stillborn babies, the sex of newborn baby and the abnormal outer table were inspected. For newborn babies, weight is measured for each individual on the day of birth and on day 4 of fertility and the birth rate [(number of newborn infants / implantation number) × 100], the stillbirth rate [(the number of stillborn infants / the number of births) × 100] and the survival rate on 4th day of newborn baby [(number of births on 4th day of clearance / number of newborns) × 100] were calculated.
Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
In males, in the group of 20 mg / kg or less, there was no death and no change was observed in the general condition.
In females, two subjects in the 20 mg / kg group and 7 subjects in the 100 mg / kg group were autopsyed on the way because all the children died by 3 rd day nursing.
Dermal irritation (if dermal study):
not examined
Mortality:
mortality observed, non-treatment-related
Description (incidence):
In males, one patient in the 100 mg / kg group had decreased spontaneous locomotor activity and slow respiration before administration on 18th administration, and died before administration on 21th administration.
In females, one patient in the 100 mg / kg group died before administration on the 22nd day of pregnancy. In addition, in another one of the 100 mg / kg group, subcutaneous carcinoma of the chest was observed from 9th pregnancy to the autopsy day (4th day of clearance). No deaths occurred in the group of 20 mg / kg or less, no change was observed in the general condition.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
In male, suppression of body weight increase or tendency was observed throughout the administration period in the group of 20 mg / kg or more, and the total body weight gain during the administration period showed a significant low value.
In females, increase inhibition was observed on days 4 and 8 during the administration period before the mating in the 100 mg / kg group. In addition, during pregnancy and rest period, suppression of body weight gain or tendency was observed in the group of 20 mg / kg or more.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
In males, there was a decrease in the early stage of administration in the group of 20 mg / kg or more and then recovered, but again decreased in the latter stage of administration after the end of the mating period.
In females, there was a decrease in the early stage of administration in the group of 20 mg / kg or more, and then recovered, but eyelid growth.
It decreased again on the day. Incidentally, in the 100 mg / kg group, an increase was observed at the administration day 8 15 days.
In addition, as an accidental change without dose correlation, increase in pregnancy 21 days was observed in the 4 mg / kg group.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
Red blood cell count and hematocrit decreased and MCH and MCHC increased in the 100 mg / kg group. In the 100 mg / kg group, the number of reticulocytes also increased. Increase in MCH was also observed in the 20 mg / kg group.
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
Increases in albumin, A / G ratio, GOT, GPT, total cholesterol, phospholipids and total bilirubin and decreases in triglyceride, sodium and potassium were observed in the 100 mg / kg group. In addition, reduction of GPT, alkaline phosphatase and creatinine was observed in the 20 mg / kg group, but both were within the physiological variation range.
Urinalysis findings:
not examined
Behaviour (functional findings):
no effects observed
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
In males, the absolute and relative weight of the thymus decreased or decreased in the 100 mg / kg group, and the absolute and relative weights of the liver and accessory kidney increased or increased. In addition, in males, the absolute weight loss of the tiles was reduced in the 100 mg / kg group, and in the 20 mg / kg group, the relative weights of the lungs, kidneys, adrenal glands, testis and epididymis were increased, 100 mg / kg Increase in relative weight of brain, heart, lung, kidney, testis and epididymis in the group
In females, an increase in the relative weight of the brain was observed in the 100 mg / kg group.
In addition, as an incidental change without dose correlation, the absolute weight increase of the adrenal gland was observed in males in the 4 mg / kg group, and in the females in the 20 mg / kg group, the absolute and relative weights of the heart were observed .
Gross pathological findings:
not specified
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Hypertrophy of central leukocytes was observed in 5 males and 7 females in the 100 mg / kg group. In males, testicular atrophy was seen in epididymal spermatogranuloma and 2 cases in control group in 1 '1 and 3 cases in control group, 4 and 100 mg / kg group, respectively, and testicular atrophy was observed In one of the cases, reduction of sperm in the epididymal cavity and ce lld ebr i S was also observed. In females, one case of 100 mg / kg group showed proliferation of transitional epithelium of kidney and papilla, in which basophilic renal tubular epithelium basification and adenocarcinoma of mammary gland were each observed in each case It was done. In females, atrophy of the thymus was observed in 1, 3 and 1 cases of control group, 20 and 100 mg / kg group, respectively.
In the case of death, one males in the 100 mg / kg group treated with neutrophilic cell infiltration and granulation, neutrophilic cell infiltration and granulation, glandular erosion, edema of the lungs, atrophy of the tiles, Gland atrophy and out Blood, female erosion of the gland stomach, edema of the lung, atrophy of the tile and necrosis of the adrenal gland were observed.
Histopathological findings: neoplastic:
not specified
Number of abortions:
no effects observed
Description (incidence and severity):
Regarding the reproductive function of the parent animal, there was no effect of administration of the test substance on the sexual cycle, mating rate, attendant fetal rate, number of days required for mating, number of corpus luteum and number of implantation traces. In addition, the influence of test substance administration was not observed in gestation period, number of births and birth rate.
Pre- and post-implantation loss:
no effects observed
Total litter losses by resorption:
no effects observed
Early or late resorptions:
no effects observed
Dead fetuses:
no effects observed
Changes in pregnancy duration:
no effects observed
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): no effects observed
Changes in number of pregnant:
no effects observed
Key result
Dose descriptor:
NOAEL
Effect level:
100 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
body weight and weight gain
food consumption and compound intake
Remarks on result:
other: males
Key result
Dose descriptor:
NOAEL
Effect level:
4 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
body weight and weight gain
food consumption and compound intake
Remarks on result:
other: females
Fetal body weight changes:
effects observed, treatment-related
Description (incidence and severity):
No abnormality was found in each group in the neonatal exterior examination as well.
In the examination of the soybean season, in 2 cases in the 20 mg / kg group and 7 cases in the 100 mg / kg group, poor behavior such as regrowth, lactation, warming and the like of children was observed, and in these mother animals All the children died, and in the group of 20 mg / kg or more, the survival rate of the newborn was decreased 4 days. Furthermore, in the 100 mg / kg group, both sexes showed a low value on the 4th day of the newborn's shooting.

Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): effects observed, treatment-related
Reduction in number of live offspring:
effects observed, non-treatment-related
Description (incidence and severity):
Furthermore, in the 100 mg / kg group, the increase in the mortality rate, the associated decrease in the fertility rate, and the decrease in female and male neonatal weight were observed. In addition, there was an increase in stillbirth rate in the 4 mg / kg group. However, similar changes were not observed in the 20 mg / kg group, so it was considered to be an accidental change.
Changes in sex ratio:
no effects observed
Description (incidence and severity):
There were no differences in the pregnancy period, the number of corpus luteums, the number of implantation traces, the number of births, the number of newborn babies, the birth rate and neonatal sex ratio in each group compared with the control group.
Changes in litter size and weights:
no effects observed
Changes in postnatal survival:
no effects observed
External malformations:
no effects observed
Skeletal malformations:
no effects observed
Visceral malformations:
not examined
Key result
Dose descriptor:
NOAEL
Effect level:
20 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
not specified
Basis for effect level:
fetal/pup body weight changes
Key result
Abnormalities:
not specified
Key result
Developmental effects observed:
not specified
Conclusions:
The NOAEL of developmental toxicity was considered to be 20 mg/kg/day, based on decreases of birth index and body weight of pups.
Executive summary:

In the OECD combined repeat dose and reproductive/ developmental (one generation)toxicity screening test [OECD TG 422], this substance was given for 49 days from 14 days before mating in males and from 14 days before mating to day 3 of lactation in females. At the 100 mg/kg group, one female died in gestation and another female was not impregnated. Birth index was decreased with increase in stillborns at the 100 mg/kg. All pups of two dams at the 20 mg/kg and seven dams at 100 mg/kg died due to the lack of nursing activity, and the viability index on day 4 after birth was consequently decreased in these groups. The body weights of pups were also decreased at birth and day 4 of lactation in the 100 mg/kg group. The decrease of litter size observed at the 100 mg/kg seems to be the chemical-induced effect although it is not statistically significant. No malformations or variations were observed in the pups.

Effect on developmental toxicity: via oral route
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEL
20 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Quality of whole database:
Two studies are available covering this endpoint.
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available
Additional information
Justification for selection of Effect on developmental toxicity: via oral route:
This study (screening study) is considered applicable and reliable. It provides the most sensitive Effect level.
The second study (Prenatal-developmental toxicity) supports the result of the selected study and results in a similar effect level.

Justification for classification or non-classification

The Data is conclusive therefore for fertility and development toxicity Benzoguanamine a classification in accordance to Regulation (EC) No 1272/2008 is not necessary.