2-methyl-4-phenylpentanol

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:

study conducted between 11th September and 19th October 1987

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: One OECD TG 407 guideline study under GLP available.

Cross-reference

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1988

Report date:

1988

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

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

Deviations:

no

Principles of method if other than guideline:

The study is similar to the OECD guidelines, however there are some differences: the dose ranges should increase between a 2 and 4 fold increase between each concentration however in this study there is nearly a ten fold increase between the intermediate and highest test concentration range.

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Pamplefleur

IUPAC Name:

Pamplefleur

Test material form:

other: clear liquid

Details on test material:

The test substance was stored under ambient conditions in non-continuous artificial light .

Test animals

Species:

rat

Strain:

Crj: CD(SD)

Sex:

male/female

Details on test animals or test system and environmental conditions:

A total of 49 rats (25 male and 24 females) of the Charles River Crl : CD (SD) BR strain was obtained from Charles River (UK) Ltd., Margate, Kent, England on 2nd September 1987 . The rats were 28 ± 1 days old, in a weight range of 67 to 86 g .

All rats were caged in groups of 5, according to sex. They had free access to tap water and Labsure LAD 1 diet. There was no information available to indicate that any substance likely to influence the effect of the test substance could reasonably be expected to be present in the diet or drinking water.

The rats were housed in Building R17, Room 3 . Animal room temperature was controlled at a mean maximum of 21.4°C and a mean minimum of 19.0°C . Relative humidity was not controlled but was recorded daily, using a wet and dry bulb thermometer at a mean of 71.3% RH. Air exchange was
maintained at a rate of approximately 15 air changes per hour . Lighting was controlled to give 12 hours artificial light in each 24-hour period.

A seven day acclimation period was allowed between delivery of the animals and the start of treatment.

Two days before the start of treatment each animal was weighed and forty rats were randomly allocated to four groups, each consistinq of five males and five females, using a computer program

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

Method of administration:
Gavage
PREPARATION OF DOSING SOLUTIONS:
The test substance was formulated freshly each day. The high (10% w/v), intermediate (1.1 % w/v) and low (0.1% w/v) dosage concentrations were prepared by direct dilution of the test substance with the vehicle (corn oil).

DIET
Analyses were made on all batches of diet used to establish levels of basic nutrients and of specified substances and micro-organisms likely to be present in feed components and which, if in excess of specified amounts , miqht have had an undesirable effect on the test system . Although occasional sliqht deviation may have been permitted, batches of diet conformed with the acceptable standards agreed by the Study Director and Head, HRC Department of Quality Assurance.

VEHICLE
- Justification for use and choice of vehicle (if other than water):
Corn oil was used

- Concentration in vehicle:
not stated

- Amount of vehicle (if gavage):
5 ml/kg/day

- Lot/batch no. (if required):
Not applicable

- Purity:
Not applicable

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The concentration of test material formulations was determined by high performance liquid chromatography.

The HPLC analysis shows that all mean results were within 5% of nominal concentrations.

Duration of treatment / exposure:

Test duration: 28 days

Frequency of treatment:

Dosing regime: 7 days/week

Doses / concentrations

Remarks:

Doses / Concentrations:
dose levels of 0, 5, 55 and 500 mg/kg/day
Basis:
actual ingested

No. of animals per sex per dose:

Male and Female: 5 animals per sex at 0 mg/kg/day (control)
Male and Female: 5 animals per sex at 5 mg/kg/day
Male and Female: 5 animals per sex at 55 mg/kg/day
Male and Female: 5 animals per sex at 500 mg/kg/day

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale:
Each animal received a constant dosage level based upon its most recently recorded bodyweight.

- Rationale for animal assignment (if not random):
Two days before the start of treatment each animal was weighed and forty rats were randomly allocated to four groups, each consistinq of five males and five females, using a computer program. so that the weight distribution within each group was similar and the initial group mean bodyweights were approximately equalised.

Positive control:

None

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule:
All animals were checked early in each working day and again in the late afternoon to look for dead or moribund animals

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule:
All animals were checked early in each working day and again in the late afternoon to look for dead or moribund animals. All animals were observed daily for signs of ill health, behavioural changes or toxicosis .

BODY WEIGHT: Yes
- Time schedule for examinations:
All rats were weighed prior to dosing and subsequently at weekly intervals throuqhout the study.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
The quantity of food consumed in each cage was measured at weekly intervals throughout the study .

FOOD EFFICIENCY: Not investigated

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
- Time schedule for examinations:
Water intake was visually observed daily except for Week 3 when gravimetric measurement was employed.

OPHTHALMOSCOPIC EXAMINATION: No data:

HAEMATOLOGY: Yes
- Time schedule for collection of blood:
At the end of the treatment period (Day 28). Blood was withdrawn under light ether anaesthesia from the orbital sinus of all surviving rats prior to termination (week 4)
- Animals fasted:
Yes - overnight
- Parameters examined.
Routine haematological parameters were examined.

BIOCHEMISTRY: Yes
- Time schedule for collection of blood: week 4

- Animals fasted: Yes
- How many animals: all surviving animals (in week 4)

- Parameters checked: Routine biochemical parameters were examined.
Some of the examinations conducted are listed below, these were all conducted using a Roche Cobas centrifugal analyser:
- Glucose
- Alkaline phosphatase (AP)
- Glutamic-pyruvic transaminase (GPT)
- Cholesterol (Chol)
- Urea nitrogen
- Total Protein

URINALYSIS: No data

NEUROBEHAVIOURAL EXAMINATION: No

Sacrifice and pathology:

EXAMPLE:
GROSS PATHOLOGY: Yes (see tables)
Rat no 18 led, which was found dead on Day 10, was autopsied and tissues were examined histologically. After 28 days of treatment (Day 29) all remaining animals were randomly killed by carbon dioxide asphyxiation and a complete autopsy undertaken . The macroscopic appearance of the tissues was recorded.

HISTOPATHOLOGY: Yes

The following organs from each animal killed after four weeks were dissected free of fat and weighed:
adrenals
ovaries
kidneys
liver
testes ( with epididymides)

Other examinations:

The following were also investigated:
Mortality data
Organ weights

Statistics:

All statistical analyses were carried out separately for males and females. The following sequence of statistical teste was used for bodyweight ,
organ weight and clinical pathology data:

I f the data consisted predominantly of one particular value (relative frequency of the mode exceeds 75% ), the proportion of values different from the mode was analysed by appropriate methods, otherwise:

Bartlett's test ( 1 ) was applied to test for heterogeneity of variance between treatments. Where significant ( at the 1% level) heterogeneity was found, a logarithmic transformation was tried to see if a more stable variance structure could be obtained.

If no significant heterogeneity was detected (or if a satisfactory transformation was found), a one-way analysis of variance was carried out.

Analyses of variance were followed by Student's 't' test and Wiliams' test ( 2 ) for a dose-related response, although only the one thought more appropriate for the response pattern observed was reported .

Results and discussion

Results of examinations

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

One male rat (no.18 ) receivinq Pamplefleur, 500 mg/kg/day was found dead following dosing on Day 10. Instances of increased salivation following dosing were observed for all rats receiving Pamplefleur, but particularly at the higher dose groups.

Mortality:

mortality observed, treatment-related

Description (incidence):

One male rat (no.18 ) receivinq Pamplefleur, 500 mg/kg/day was found dead following dosing on Day 10. Instances of increased salivation following dosing were observed for all rats receiving Pamplefleur, but particularly at the higher dose groups.

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

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):

In comparison with control rats, statistically significantly lower (P < 0.05 ) haemoglobin levels and red blood cell counts were recorded for female rats receiving Pamplefleur, 500 mg/kg/day .

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

In the 500 mg/kg/bw dose range

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

Statistically significantly hiqher (P<0.01 ) adjusted liver weiqhts were recorded for both male and female rats receiving Pamplefleur, 500 mg/kg/day in comparison with controls .

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Histopathological findings: neoplastic:

effects observed, treatment-related

Details on results:

Increased salivation following dosing, which was observed for all rats receiving Pamplefleur, was recorded more frequently for rats in the higher dosage groups . Increased salivation prior to treatment was also observed for female rats in the high dosage groups receiving Pamplefleur, 500 mg/kg/day. These clinical signs were not observed for control rats.

Mortalities:
One male rat receiving Pamplefleur, 500 mg/kg/day was found dead following dosing on Day 10 . Instances of increased salivation following dosing were observed for this animal prior to death and at macroscopic pathology the lungs were not collapsed. Histological examination revealed minimal congestion of the lungs; the cause of death of this animal was not established.

Body weight changes:
No changes in bodyweight gain were recorded during the four-week treatment period that were considered to be related to treatment with Pamplefleur .

Food consumption
For male rats receiving Pamplefleur, 500 mg/kg/day, sliqhtly lower food consumption was recorded during Week 1 in comparison with controls . However, during Weeks 2 , 3 and 4, food consumption for these animals was similar to that of the controls and no changes in food consumption were recorded during the four-week treatment period for the remaining rats receiving Pamplefleur.

Haematology:
In comparison with control rats, statistically significantly lower (P < 0.05 ) haemoglobin levels and red blood cell counts were recorded for female rats receiving Pamplefleur, 500 mg/kg/day . Haemoglobin levels for female rats in the intermediate dosage group (Pamplefleur, 55 mg/kg/day) were also siqnificantly lower (P<0.05 ) than those of the controls . Individual haemoglobin levels for female rats receiving Pamplefleur , 55 or 500 mg/kg/day, were, however, within the expected parameter range .

No other haematological changes were observed that were considered to be related to treatment with Pamplefleur.

Clinical Chemistry:
Glutamic-pyruvic transaminase (GPT) levels for female rats receivinq Pamplefleur, 500 mg/kg/day were statistically significantly hiqher (P<0.05) than those of rats receiving corn oil.

Calcium levels for both male and female rats receiving Pamplefleur in the 55 or 500 mg/kg/day were lower than those of the controls with statistical significance (P<0.05 ) being achieved for male and female rats in the high dosage group and for male rats in the intermediate dosage group.

In comparison with rats receiving corn oil, statistically significantly higher (P<0.05 ) urea nitrogen levels were recorded for male rats receiving Pamplefleur, 500 mg/kg/day.

Potassium levels for female rats receiving Pamplefleur were higher than those of the controls with statistical significance ( P<0.05 ) being achieved for female rats receiving Pamplefleur, 55 or 500 mg/kg/day.

Organ weights:
Statistically significantly higher (P<0.01 ) adjusted liver weiqhts were recorded for both male and female rats receiving Pamplefleur 500 mg/kg/day in comparison with those receiving corn oil. Adrenal, testes and ovary weights for rats receiving Pamplefleur were similar to those of the controls receiving corn oil .

MACROSCOPIC PATHOLOGY:
Enlargement of the liver was observed for two male rats (no . 16 and 19 ) receiving Pamplefleur , 500 mg/kg/day.
Hair lose from the dorsal surface of the skin was observed for rat no . 174 receiving Pamplefleur, 500 mg/kg/day. Hair loss from the dorsal surface and from the forelimbs was similarly observed for rat no . 20 receiving Pamplefleur, 500 mg/kg/day.

For three female rats (no. 36 , 37 and 38 ) receiving Pamplefleur, 500 mg/kg/day brown staining of the fur on the dorsal surface was recorded at termination. The above abnormalities of the skin (hair loss) and fur (stained) were not observed for control rats receiving corn oil .

MICROSCOPIC PATHOLOGY:
Treatment-related finding in the 500 mg/kg/day dose group:
Kidneys:
Minimal/moderate cortical tubular eosinophilic inclusions in two male rats.

Livers:
Apparent minimal generaIised hepatocyte enlargement in all the female rats and in two male rats .

No treatment-related findings were detected in rats receiving 5 mg/kg/day or 55 mg/kg/day dose groups.

All other findings were considered to be spontaneous in origin and of no toxicological importance .

Effect levels

Dose descriptor:

NOAEL

Effect level:

55 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Target system / organ toxicity

Critical effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

The 'no adverse effect' level for oral administration of Pamplefleur to rats was considered to be 55 mg/kg/day.

Executive summary:

In and OECD TG 407 Pamplefleur, formulated in corn oil , was administered to rats by intragastric intubation at dosaqe levels of 5 , 55 or 500 mg/kg/day. Treatment was carried out once daily, for twenty-eight consecutive days. Control animals similarly received corn oil at a dosage volume of 5 ml/kg/day.

Clinical findings:

Increased salivation following dosing, which was observed for all rats receiving Pamplefleur, was recorded more frequently for rats in the higher dosage groups . Increased salivation prior to treatment was also observed for female rats in the high dosage groups receiving Pamplefleur, 500 mg/kg/day. These clinical signs were not observed for control rats.

Mortalities:

One male rat receiving Pamplefleur, 500 mg/kg/day was found dead following dosing on Day 10 . Instances of increased salivation following dosing were observed for this animal prior to death and at macroscopic pathology the lungs were not collapsed. Histological examination revealed minimal congestion of the lungs; the cause of death of this animal was not established.

Food consumption

For male rats receiving Pamplefleur, 500 mg/kg/day, sliqhtly lower food consumption was recorded during Week 1 in comparison with controls . However, during Weeks 2 , 3 and 4, food consumption for these animals was similar to that of the controls and no changes in food consumption were recorded during the four-week treatment period for the remaining rats receiving Pamplefleur.

Haematology:

In comparison with control rats, statistically significantly lower (P < 0.05 ) haemoglobin levels and red blood cell counts were recorded for female rats receiving Pamplefleur, 500 mg/kg/day . Haemoglobin levels for female rats in the intermediate dosage group (Pamplefleur, 55 mg/kg/day) were also siqnificantly lower (P<0.05 ) than those of the controls . These decreases were ca 5% compared to the controls. Individual haemoglobin levels for female rats receiving Pamplefleur , 55 or 500 mg/kg/day, were, however, within the expected parameter range.

Biochemistry:

Higher albumin and lower globulin levels, resulting in higher albumin/globulin (A/G) ratios, were recorded for male rats receiving Pamplefleur in comparison with those receiving corn oil . Statistical significance was achieved in most instances (P<0.05 or P<0.0l ) and the observed changes were considered to be related to treatment in male rats receiving Pamplefleur, 500 mg/kg/day, though the effects were only minor < 10% compared to the controls. The changes recorded in serum protein levels in male rats receiving Pamplefleur, 55 mg/kg/day were small in magnitude and were not considered to be of toxicological importance.

In comparison with control rats, statistically significantly higher (P<0.05 ) glutamic-pyruvic transaminase (GPT) levels were recorded for female rats receiving Pamplefleur, 500 mg/kg/day around 26% . A similar trend (< 20% increase) higher glutamic-oxaloacetic transaminase (GOT) levels was observed for female rats receiving Pamplefleur although statistical significance was not achieved. The shifts in GPT and GOT levels were not observed for male rats.

Calcium levels for both male and female rats in the intermediate and high dosage groups were lower than those of the controls, with statistical significance being achieved (P<0.05) for male and female rats in the high dosage group and for male rats in the intermediate dosage group. The decrease is < 5% and therefore the toxicological relevance is not clear.

In comparison with rats receiving corn oil , statistically significantly higher ( P<0.05) urea nitrogen levels (maximally 16%) were recorded for male rats receiving Pamplefleur, 500 mg/kg/day. No similar trends were apparent for female rats .

Organ weight analysis:

Statistically significantly hiqher (P<0.01) adjusted liver weights were recorded for both male and female rats receiving Pamplefleur, 500 mg/kg/day in comparison with controls . Similarly, kidney weights for male (unadjusted weights) and female (adjusted weights) rats receiving Pamplefleur were generally higher than those of the controls, with statistical significance being achieved for female rats receiving Pamplefleur, 55 or 500 mg/kg/day (P<0.05 and P<0.01 respectively).

Macroscopic pathology

Pale, mottled and enlarged kidneys were observed at termination amongst male and female rats receivinq Pamplefleur, 55 or 500 mg/kg/day. Enlargement of the liver was also observed for two male rats receivinq Pamplefleur, 500 mg/kg/day. These macroscopic abnormalities were not observed for control rats receiving corn oil and were therefore considered to be related to treatment with Pamplefleur.

Hair loss from the forelimb and/or dorsal surface was noted for two male rats receivinq Pamplefleur, 500 mg/kg/day while brown staining of the fur on the dorsal surface was noted for three female rats in this dosage group . Neither hair loss nor brown staining of the fur were observed for control rats or for ras receiving Pamplefleur, 5 or 55 mg/kg/day.

Microscopic pathology:

Histological examination revealed minimal to moderate eosinophilic inclusions in the kidneys of two male rats treated with Pamplefleur, 500 mg/kg/day. In the livers, apparent minimal generalised hepatocyte enlargement was present in all female rats and in two male rats receiving Pamplefleur, 500 mg/kg/day . These findings were considered to be related to treatment. No changes were apparent in the livers or kidneys of rats receiving Pamplefleur, 55 mg/kg/day.