[No public or meaningful name is available]

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

March - May 2019

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2019

Report date:

2019

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

2,6-bis[[bis(2-hydroxyethyl)amino]methyl]-4-nonylphenol

EC Number:

243-500-6

EC Name:

2,6-bis[[bis(2-hydroxyethyl)amino]methyl]-4-nonylphenol

Cas Number:

20073-51-2

Molecular formula:

C25H46N2O5

IUPAC Name:

2,6-bis{[bis(2-hydroxyethyl)amino]methyl}-4-nonylphenol

Test material form:

liquid: viscous

Test animals

Species:

rat

Strain:

Wistar

Details on species / strain selection:

RccHan: WIST. Female, Nulliparous and non pregnant. 8-9 weeks old

Sex:

female

Administration / exposure

Route of administration:

oral: gavage

Details on route of administration:

The route of vehicle and test item administration was oral through gavage and was selected after discussion with the sponsor.

Vehicle:

corn oil

Duration of treatment / exposure:

The duration of dosing was for 14 days. The first day of dose administration was designated as day 1 for each rat.

Frequency of treatment:

Dose formulation was administered once daily by oral gavage at approximately the same time each day using intubation cannula attached to a graduated syringe.

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

250 mg/kg bw/day (nominal)

Doses / concentrations 2

Dose / conc.:

500 mg/kg bw/day (nominal)

Doses / concentrations 3

Dose / conc.:

1 000 mg/kg bw/day (nominal)

No. of animals per sex per dose:

5 animals of each sex at each dose.

Control animals:

yes

Positive control:

No positive control

Examinations

Observations and examinations performed and frequency:

All rats were observed twice a day for visible clinical signs during treatment period.

Sacrifice and pathology:

All surviving rats were fasted overnight (with ad libitum supply of drinking water) prior to sacrifice. At terminal sacrifice and morbidity, rats were ehtanized by carbon dioxide asphyxiation and subjected to a full gross necropsy under the direct supervision of a veterinary pahtologist. Rats, which were found dead or sacrificed under moribund condition, were subjected to post-mortem examination. Rats were examined carefully for external abnoramlities. The thoracic and abdominal cavities were opened and a thorough examination of the argnas was carried out to detect any abnormalities.

Other examinations:

Food consumption was measured daily.
All organs were weighed from all surviving rats.

Results and discussion

Results of examinations

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

All rats belonging to vehicle control group were found normal throughout the treatment period. The observed clinical signs were:
weakness, polyuria,piloerection, salivation,

Mortality:

mortality observed, treatment-related

Description (incidence):

Mortality and morbidity
No morbidity and mortality was observed in rats from vehicle control and low dose groups.
Three mortalities were observed in high dose group. Two mortalities qwere observed in mid dose group.
Three male and four female rats were sacrificed under moribund condition in high dose group. Three male and five female rats were sacrificed udner moribund condition in mid dose group.
The rats sacrificed under moribund condition and mortality observed in mid and high dose groups were considered as treatment related as the mortality observed after showing severe clinical sign.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Statistically significant decrease in mean body weight was observed in male and female rats of low dose group on treatment days 8, 11 and 14 as compared with that of the control group.

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

Statistically significant reduction in mean food consumption in low dose group was observed in male rats during treatment days 8-11 and in female rats during treatment days 4-8 and 1-14 as compared with that of the control gorup. Similar reduciton in mean food consumption without statistical significance in low dose group was observed in male rats during treatment days 1-4, 4-8, 11-14, and 1-14. $Reduction i9n food consumption was considered as weffect of test item as concurrent decrease ind boyd weight and body weight change was observed.

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

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

In male rats, statistically significant decrease in terminal body weight and absolute weight of thymus, prostate, and Levator ani plus bulbocavernosus muscles was observed in low dose group.
Statistically signifcant increase in relative weight of brain and testes and statistically significant decrease in relative weight of prostate was observed in low dose group as compared with that of the control group.
Increase in relative weight of brain and testes could be correlated with decreased terminal body weight. Whereas decreased in absolute weight of thymus, prostate and LABC and relative weight of rpostate could be consdiered as effect of test item.
In female rats, statistically signifcant decrease in terminal body weight and increase in absolute weight of spleen was observed in low dose group as compared with that of the control group. Statistically significant increase in relative weight of spleen and brain was observed in low dose group as compared with that of the control group.
Increase in relative weight of brain could be correlated with decreased terminal body weight, whereas increased in absolute and relative weight of spleen could be considered as effect of test item.
Decreased in terminal body weiht was considered as effect of test item as it could be correlated with decreasesed body weight adn food consumption.

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

External examination of found dead, moribundly sacrificed, and termianlly sacrificed rats of either sex belonging to all groups did not revaeal any abnormality.

Internal examination of found dead male rats revelaed gaseous distension of stomach (in 3 of 4 rats) gaseous distension of itnestine (1in 4 rats) and whitish foci - non glandular stomach (in 1 of 4 rats) whereas internal examination of found dead female rat revealed gaseous distension of stomach.
Internal examination of moribundly sacrificed male reats revealed gaseous distension of stomach (in 2 of 6 rats), whitish foci - non glandular stomach (in 6 of 6 rats), and reddish foci -non glandular stoamch (in 1 o f 6 rats) whereas internal examination of moribundly sacrifcie female rats revelaed gaseous distension of stomach (in 5 of 9 rats) and whitish foci -n on glandular sotamch (in 8 of 9 rats).
Internal examination of terminally sacrificed rats of either sex belonging to vehicle control and low dose groups did not reveal any abnormality.
Changes observed during internal examination of found dead and moribundly sacrificed rats were considered as effects of test item.

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

External examination of found dead, moribundly sacrificed, and termianlly sacrificed rats of either sex belonging to all groups did not revaeal any abnormality.

Internal examination of found dead male rats revelaed gaseous distension of stomach (in 3 of 4 rats) gaseous distension of itnestine (1in 4 rats) and whitish foci - non glandular stomach (in 1 of 4 rats) whereas internal examination of found dead female rat revealed gaseous distension of stomach.
Internal examination of moribundly sacrificed male reats revealed gaseous distension of stomach (in 2 of 6 rats), whitish foci - non glandular stomach (in 6 of 6 rats), and reddish foci -non glandular stoamch (in 1 o f 6 rats) whereas internal examination of moribundly sacrifcie female rats revelaed gaseous distension of stomach (in 5 of 9 rats) and whitish foci -n on glandular sotamch (in 8 of 9 rats).
Internal examination of terminally sacrificed rats of either sex belonging to vehicle control and low dose groups did not reveal any abnormality.
Changes observed during internal examination of found dead and moribundly sacrificed rats were considered as effects of test item.

Histopathological findings: neoplastic:

no effects observed

Other effects:

not specified

Effect levels

Key result

Dose descriptor:

LOAEL

Effect level:

ca. 250 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

body weight and weight gain

clinical signs

food consumption and compound intake

Target system / organ toxicity

Key result

Critical effects observed:

yes

Lowest effective dose / conc.:

250 mg/kg bw/day (nominal)

System:

other: lymphatic system

Organ:

thymus

Treatment related:

yes

Dose response relationship:

yes

Relevant for humans:

not specified

Applicant's summary and conclusion

Conclusions:

Based on the results of present study, under the condition and procedures followed, dose levels of 1000 and 500 mg /kg b.wt. produced severe toxictiy viz. treatment related mortality, morbidity, and adverse clinical signs whereas a dose level of 250 mg/kg b. wt. porduced mild toxicity viz. decreased body weight change (7.90% in males), food consumption and treatment related changes in absolute and relative organ weight. Hence, following dose levels are suggested for reproduction/developmental toxicity screening study (dose range finding study):
Option Low dose Mid dose High dose (mg/kg)
I 15 50 150
II 15 60 240
III 20 60 180

Executive summary:

EXECUTIVE SUMMARY: The study was conducted to determine the adverse effects occurring as result of repeated daily administration of the POLIOL MB 600 in Wistar rat for a period of 14 consecutive days and to select the dose levels for further reproductive/developmental toxicity screening study. 

Method: 

 

Test Item 

POLIOL MB 600  

Test System 

Wistar strain (RccHan:WIST) 

Route of Administration 

Oral through gavage 

Dose Levels (mg/kg b. wt./day) 

G1 (vehicle control): 0; G2 (low dose): 250; G3 (mid dose): 500; G4 (high dose): 1000 

Duration of Treatment 

14 days 

N° of Rats 

5 rats/sex/group 

Frequency of Dose Formulation Preparation 

Daily fresh preparation 

 

All rats were observed twice a day for mortality, morbidity and visible clinical signs throughout the 
treatment period. Body weight of all surviving rats was recorded on days 1, 4, 8, 11, 14 and 15. Food 

consumption was calculated during treatment period for all surviving rats. At terminal sacrifice, surviving 
rats were sacrificed by carbon dioxide asphyxiation and subjected to gross pathological examination. 

Absolute organ weights for all surviving rats were recorded and relative organ weights were calculated for 
the organs: liver, kidneys, adrenals, testes, epididymis, prostate, LABC, seminal vesicle with coagulating 

gland, thymus, heart, brain, spleen, uterus with cervix, and ovaries.  

 Results: 

POLIOL MB 600 at 250 mg/kg b. wt. /day 

No mortality was observed. 

Salivation was observed in all male and female rats from treatment day 8. 

Treatment related decrease in body weight, body weight  change, food consumption and 
terminal body weight were observed in male and female rats. 

In male rats, statistically significant decrease in absolute weight of thymus, prostate, and LABC 
and relative weight of prostate were observed. In female rats, statistically significant increase 

in absolute and relative weight of spleen was observed. These changes in organ weights 
(absolute and relative) were considered as treatment related however adversity of effect could 

not be established due to limited end-points of this study. 

External and internal examination of rats of either sex did not reveal any abnormality.   

 

POLIOL MB 600 at 500 mg/kg b. wt. /day 

Two mortalities (two male rats) were observed. 

Three male and five female rats were sacrificed under moribund condition. 

Treatment related clinical signs viz. weakness, salivation, piloerection (only in male rats) and polyuria were observed in all male and female rats. 

Internal examination of found dead male rats revealed gaseous distension of stomach (in 1 of rats) and whitish foci – non-glandular stomach (in 1 of 2 rats). 

Internal examination of moribundly sacrifice male rats revealed gaseous distension of stomach (in 1 of 3 rats) and whitish foci – non-glandular stomach (in 3 of 3 rats). 

Internal  examination  of moribundly  sacrifice female  rats  revealed  gaseous  distension  of stomach (in 2 of 5 rats) and whitish foci – non-glandular stomach (in 5 of 5 rats). 

 POLIOL MB 600 at 1000 mg/kg b. wt. /day 

Three mortalities (2 male and 1 female rats) were observed. 

Three male and four female rats were sacrificed under moribund condition. 

Treatment related clinical signs viz. weakness, salivation, piloerection and polyuria were observed in all male and female rats. 

Internal examination of found dead male rats revealed gaseous distension of stomach (in 2 of 2 rats) and gaseous distension of intestine (in 1 of 2 rats). 

Internal examination of found dead female rat revealed gaseous distension of stomach. 

Internal examination of moribundly sacrificed male rats revealed gaseous distension of stomac (in 1 of 3 rats), whitish foci – non-glandular stomach, (in 3 of 3 rats) and reddish foci – non- 

glandular stomach (in 1 of 3 rats). 

Internal examination of moribundly sacrificed female rats revealed gaseous distension of stomach (in 3 of 4 rats) and whitish foci – non-glandular stomach, (in 3 of 4 rats). 

In male rats, statistically significant decrease in absolute weight of thymus, prostate, and LABC 
and relative weight of prostate were observed. In female rats, statistically significant increase 

in absolute and relative weight of spleen was observed. These changes in organ weights 
(absolute and relative) were considered as treatment related however adversity of effect could 

not be established due to limited end-points of this study. 

Conclusion: 

Based on the results of present study, under the condition and procedures followed, dose levels of 1000 and 

500 mg/kg b. wt. produced severe toxicity viz. treatment related mortality, morbidity, and adverse clinical 

signs whereas a dose level of 250 mg/kg b. wt. produced mild toxicity viz. decreased body weight change 

(7.90% in males), food consumption and treatment related changes in absolute and relative organ weight. 

Hence, following dose levels are suggested for reproduction/ developmental toxicity screening study (dose 

range finding study): 

Options              Low dose              Mid dose              High dose

I                     15                                 50                     150

II                    15                                 60                     240

III                   20                                60                    180 

(mg/kg b. wt/day)