1-chloro-N,N-diethyl-1,1-diphenyl-1-(phenylmethyl)phosphoramine

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:

From February 1990 to April 1991

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study performed according to a standardized method and in compliance with GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Report date:

1991

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

GM102E

IUPAC Name:

GM102E

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

batch no. 8724

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Italia S.p.A., Via Indipendenza 11 - 22050 Calco (Como)
- Age at study initiation: about 4 weeks (27-29 days)
- Weight at study initiation: males = 75-85 g; females = 60-70 g.
- Fasting period before study: none
- Housing: in a room with limited access, in makrolon cages measuring 42x26x16 cm, each fitted with a stainless steel cover-feed rack. Dust-free poplar/fir chips, heat processed for resin removal, were used for bedding. The animals of each group were caged in 2 cages/sex of 3 or 2 animals/cage.
- Diet (e.g. ad libitum): on a dry matter basis (moisture 12%), the diet contained crude proteins (18.5%), crude fat (3%), crude fiber (6%) and ash (7%). This was supplemented by vitamins and trace elements.
- Water (e.g. ad libitum): municipal filtered water was distributed ad libitum by a means of an automatic wetring valve system.
- Acclimation period: 2 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/-2°C
- Humidity (%): 60 +/-20%
- Air changes (per hr): 12
- Photoperiod (hrs dark / hrs light): 12-hour circadian cylce (7 a.m. - 7 p.m.)

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: Deionized water

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:
Every day or two days suitable amounts of test article water solutions were prepared in order to obtain the final concentrations of 0.625, 1.25, 2.5 and 5 mg/mlto be administered, respectively, to the animals at a constant administration volume of 10 ml/kg b.w.
The control animals received 10 ml/kg/day of deionized water.
The volumes to be administered were calculated on the basis of the most recently recorded body weight (individual values).

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

A stability of the test article water solutions was ascertained for 48 hours.
The concentration of the test article water solutions was checked one during the treatment period: at week 4 of the study.

Duration of treatment / exposure:

Test duration: 28 days

Frequency of treatment:

Dosing regime: once a day, 7 days/week

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 6.25, 12.5, 25 and 50 mg/kg bw/day
Basis:
nominal in water

No. of animals per sex per dose:

5 animals per sex and per dose.

Control animals:

yes, concurrent vehicle

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily
- Cage side observations checked in table [No.?] were included.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION AND COMPOUND INTAKE :
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/animal/day: Yes
- Compound intake 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

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: before the test (day 9 for males; day 10 for females) and by the end of the dosing period (week 4: day 26 for males; day 25 for females)
- Dose groups that were examined: all animals were inspected.

HAEMATOLOGY: Yes
- Time schedule for collection of blood: during week 5, on day 29 of the study
- Anaesthetic used for blood collection: Yes (ether)
- Animals fasted: Yes
- How many animals: all
- Parameters checked in table [No.?] were examined.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: during week 5, on day 29 of the study
- Animals fasted: Yes
- How many animals: all
- Parameters checked in table [No.?] were examined.

URINALYSIS: Yes
- Time schedule for collection of urine: during week 5, on day 29 of the study
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes
- Parameters checked in table [No.?] were examined.

NEUROBEHAVIOURAL EXAMINATION: No

Sacrifice and pathology:

GROSS PATHOLOGY: Yes (see table)
HISTOPATHOLOGY: Yes (see table)

Results and discussion

Results of examinations

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

No mortality. Salivation observed just after treatment in animals treated with the two highest doses starting from week 3 onwards.

Mortality:

mortality observed, treatment-related

Description (incidence):

No mortality. Salivation observed just after treatment in animals treated with the two highest doses starting from week 3 onwards.

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

not specified

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

not specified

Ophthalmological findings:

no effects observed

Haematological findings:

no effects observed

Description (incidence and severity):

some incidental effects observed in males but no relation to the dosages administered was found.

Clinical biochemistry findings:

no effects observed

Description (incidence and severity):

some incidental effects observed in females but not dose-related.

Urinalysis findings:

no effects observed

Behaviour (functional findings):

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

some incidental effects observed but not dose-related and in the range of controls.

Gross pathological findings:

no effects observed

Description (incidence and severity):

some incidental effects observed but not dose-related

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

some incidental effects observed but not dose-related

Details on results:

CLINICAL SIGNS AND MORTALITY
No death occured during the experimental period.
Salivation after the administration of the compound in almost all the animals of both sexes at 25 and 50 mg/kg starting from the 3rd week of the treatment.

BODY WEIGHT AND WEIGHT GAIN
Body weight of animals of both sexes was found to have been unaffected by treatment. No significant differences between control and test item were recorded at any time.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study)
No changes in food attributable to treatment were found at any dose.
Incidental but significant differences from the controls in males at 6.25 mg/kg/day (increase) and at 25 mg/kg/day (decrease). These changes occured only during week 3.

OPHTHALMOSCOPIC EXAMINATION
No eye abnormalities were observed in any of the animals.

HAEMATOLOGY
Some significant differences from the control values:
- increase in erythrocyte number and hemoglobin concentration in males tested with 12.5 and 50 mg/kg/day of test item, as well as in the hematocrit value of males tested with 50 mg/kg/day of test item.
- increase in mean Porthrombin Time in males tested exposed to 12.5 mg/kg/day of test item.
No relation to the dosages administered was found and all the individual values fell within the norm.
No statistically significant differences observed in females.

CLINICAL CHEMISTRY
Slight variations in the electrophoretic pattern were found in females exposed to 12.5 and 25 mg/kg/day of test item but they were not dose related.
An increase in total cholesterol was recored in all the dosed females but particularly low levels of total cholesterol were measured in some control females while values of the treated animals stayed within the normal range of animals of this strain and age.
A slight increase of sodium level was observed on in the animals tested with the lwoest dose (6.25 mg/kg/day). Individual data did not reveal any notable change.

URINALYSIS
Statistical analysis carried out on some parameters (diuresis, specific gravity and pH) did not reveal aby significant difference between control and dosed animals of eiher sex.
No changes were found at the semiquantitative analysis of the urine and sediment microscopy.

ORGAN WEIGHTS
A slight not statistically significant decrease with respect to controls of mean weights of spleen in males dosed with 50 mg/kg/day and of adrenal glans in females dosed with 12.5, 25 and 50 mg/kg/day was observed. This decrease is characterized by individual values weel comprised in the normal range of controls of this or other experiments performed in the same lab with rats of the same strain and age.
No dose relationship was seen in the adrenal values.
A decrease of ovary mean absolute and relative values of group tested with 12.5 mg/kg/day was observed but it was considered related to normal estrual cycle changes.

GROSS PATHOLOGY
Gastric glandular mucosa erosions were seen in one control female, in one female tested with 25 mg/kg/day of test item and in 2 females tested with 50 mg/kg/day of test item. Some other changes (watery feces, congested thymus or submandibular lymph nodes, pale liver) were seen sporadically in individual treated rats without any correlation with the treatment.
All modifications that were seen were slight aspects of incidental spontaneous pathology not infrequently seen in untreated rats of the same strain and age.

HISTOPATHOLOGY: NON-NEOPLASTIC
Some modifications were observed: liver aggregates of mononuclear cells adjacent to degenerated hepatocytes, vacuolization consistent with fatty change and periportal subacute inflammation, kidney aggregates of mononuclear cells, tubular basophilia or steatosis, presence of hyline droplets, stomach glandular mucosa erosion, submandibular lymph nodes plasmocytosis of medullary cords and erythrophagocytosis, adrenal cortex congestion and degeneration.
The modiciations were considered incidental and are not related to the the treatment.

HISTORICAL CONTROL DATA
Some spontaneous pathologies observed in this study are not infrequent in rats of the same age and strain in the laboratory. Most of them appeared in this study with the same frequency and degree in control and in trated rats.

The supplier of GM102E confirmed that the test item is stable in aqueous solution (pH 5-7).

Effect levels

1

Dose descriptor:

NOAEL

Effect level:

50 mg/kg bw/day (nominal)

Basis for effect level:

other: original NCD unit is mg/kg/day.

2

Dose descriptor:

NOEL

Effect level:

12.5 mg/kg bw/day (nominal)

Basis for effect level:

other: original NCD unit is mg/kg/day.

Target system / organ toxicity

Critical effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

In the conditions of the study, no dose-related adverse effects were seen on male and female rats after ingestion of GM102E at concentrations up to 50 mg/kg/day during 28 days. Based on the results of the study, the NOAEL for 28-day repeat dose administration of GM102E by oral route in the rat is considered to be 50 mg/kg/day while the NOEL is considered to be 12.5 mg/kg/day.

Executive summary:

The effects of a repeated oral administration of the test item GM102E were studied during 28 days on Sprague Dawley Crl:CD(SD)BR rats.

The test item was administered by gavage to rats once a day for 4 consecutive weeks. The test item was dissolved in deionized water before administration in order to obtain the following doses: 6.25, 12.5, 25 and 50 mg/kg/day and a control group was exposed only to deionized water. Each tested group consisted of 5 males and 5 females. The administration volume was kept constant at 10 ml/kg b.w. in all the groups.

At the end of the 4-week exposure period, blood collection was performed for all animals and they were killed immediately for examinations.

No animals were found dead during the study. Based on clinical observations (clinical signs observed daily, body weight observed weekly, food consumption, ophthalmoscopic observations) the only effect attributable to treatment was salivation just after dosing which was observed in almost all animals treated at 25 and 50 mg/kg/day starting from week 3 onwards.

Overall body weight gain in surviving males and females was within the range expected for rats of this strain and age used in this type of study.

Laboratory investigations (hematology, blood chemistry and urinalysis) and post-mortem examinations (gross pathology, organ weights and histology) did not reveal any treatment-related changes.

Thus, based on the results of the study, the NOAEL for 28-day repeat dose administration of GM102E by oral route in the rat is considered to be 50 mg/kg/day while the NOEL is considered to be 12.5 mg/kg/day.

Based on these results and according to the Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures (CLP), GM102E should not be classified as toxic after repeated exposure.