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Diss Factsheets

Administrative data

Endpoint:
short-term repeated dose toxicity: oral
Remarks:
combined repeated dose and reproduction / developmental screening
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
1997
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Guideline study without detailed documentation

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1997

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Deviations:
no
GLP compliance:
not specified
Limit test:
no

Test material

Constituent 1
Reference substance name:
4-hydroxy-4-methylpentan-2-one
EC Number:
204-626-7
EC Name:
4-hydroxy-4-methylpentan-2-one
Cas Number:
123-42-2
IUPAC Name:
4-hydroxy-4-methylpentan-2-one
Constituent 2
Reference substance name:
4-hydroxy-4-methyl-2-pentanone (HMP)
IUPAC Name:
4-hydroxy-4-methyl-2-pentanone (HMP)
Details on test material:
- Name of test material (as cited in study report): diacetone alcohol CAS# 123-42-2
- Physical state: liquid
- Analytical purity: 99.8%
- Stability under test conditions: stable
- Storage condition of test material: sealed in a container, in a cool, dark location

Test animals

Species:
rat
Strain:
other: SD(Crj:CD(SD)) SPF
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Japan Co (955 Kanbayashi, Yatabe-machi, Niihari-gun, Ibaragi-ken)
- Age at study initiation: 8 week old male rats and 7 week old female rats
- Weight at study initiation: 358 g for males and 211 g for females
- Housing: animals were raised in barrier system cages arranged in 5 stainless steel racks
- Diet (e.g. ad libitum): Nihon Nosan Kogyo Co., solid food lab MR, stock, available ad libitum
- Water (e.g. ad libitum): Kanagawa prefecture water supply available ad libitum
- Acclimation period: 6 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3
- Humidity (%): 40-60
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light): 12/12


Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
VEHICLE
- Vehicle: Japanese Pharmacopeia purified water
- Lot/batch no. (if required): 180889, 180964
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Since the solution was confirmed to be stable for at least 7 days, it was prepared weekly and used within 7 days. The intial and final preparations were analyzed, and confirmation was made that specific concentrations were prepared. Analysis of the raw material of the test substance was conducted.
Duration of treatment / exposure:
44 days for males and 41-45 days for females
Frequency of treatment:
daily
Doses / concentrations
Remarks:
Doses / Concentrations:
30, 100, 300, or 1000 mg/kg body weight
Basis:
actual ingested
No. of animals per sex per dose:
10 animals/sex/dose
Control animals:
yes
Details on study design:
Dose levels were selected on the basis of results from a 14-day dose-range finding study.
Positive control:
None used

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: on kill date
- Anaesthetic used for blood collection: Yes (ether)
- Animals fasted: Yes
- How many animals: all

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: on kill date
- Animals fasted: Yes
- How many animals: all

URINALYSIS: Yes
- Time schedule for collection of urine: collected on day 38 and day 41 of administration
- Metabolism cages used for collection of urine: Yes
- Animals fasted: No data
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes
Other examinations:
Reproductive examinations
Statistics:
Statistical significance (danger rate of 5% or less) with the control group on the mean values and frequencies was conducted using the following methods. Parametric data such as body weight, food consumption, hematological and blood chemical data, organ weight, number of corpus luteum, number of implantations, gestation period, and number of pups delivered were subject to Bartlett’s distribution test. If the distribution was normal, an distribution analysis for the normal positions was conducted, and if a significant variance was confirmed from those results, a comparative test was performed on each group compared with the control group using either the Dunnett method or the Scheffe method (if the size of the groups differed). If the distribution was not normal and for non-parametric data such as implantation rate, delivery rate, pregnancy rate, rate of surviving pups, and qualitative data for urine examinations, Kruskal-Wallis analysis of variance was performed, and if a significant variance was confirmed for those results, a comparative test was performed on each group compared with the control group using either the Dunnett method or the Scheffe method (if the size of the groups differed). Categorical data such as the survival rate for newborns, copulation rate, fertilization rate, birth rate, gender ratio of newborns, changes in the overall condition and manifestation of pathological abnormalities were subject to χ2 tests. Pathological abnormalities were also noted in the control group, and the data for findings where the impact of the test substance exhibited a difference in the distribution for the degree of changes was separated into two categories and tested.

Results and discussion

Results of examinations

Clinical signs:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
no effects observed
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
effects observed, treatment-related
Clinical biochemistry findings:
effects observed, treatment-related
Urinalysis findings:
no effects observed
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Gross pathological findings:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Histopathological findings: neoplastic:
no effects observed
Details on results:
CLINICAL SIGNS AND MORTALITY
There were no deaths among the males. In the females, one animal in the 1000 mg/kg group was weakened during delivery and so was euthanized once reaching a near death state. Other than this one case, no deaths were noted.

BODY WEIGHT AND WEIGHT GAIN
There were no significant changes noted in the weights or weight increases in males. In females, a significant reduction in the amount of weight gain during the premating period was noted in the 1000 mg/kg group.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study)
Neither males nor females in the groups administered the test substance exhibited significant changes in food consumption.

HAEMATOLOGY
There was a significant increase in the number of platelets noted in the 1000 mg/kg group.

CLINICAL CHEMISTRY
In the 1000 mg/kg group, significant increases in GOT, choline esterase, total protein, total cholesterol, total bilirubin, blood urea nitrogen, creatinine and calcium were noted, as well as a significant reduction in glucose. Furthermore, while not a significant difference, an increasing trend was noted in the 300 mg/kg group for choline esterase and total protein, an increasing trend was noted in the 1000 mg/kg group for γ-GTP and albumin, along with a decreasing trend for the A/G ratio and triglicerides. Significantly low values were exhibited for alkaline phosphatase when compared to the control group but these were not dose dependent changes.

URINALYSIS
There were no significant changes noted for any of the examination items in the groups administered the test substance.

ORGAN WEIGHTS
Significant increases in the absolute and relative weights of the kidneys were noted in males in the 300 mg/kg group, but these were not confirmed to be statistically significant, and the same increasing trend was noted in the 1000 mg/kg group. Additionally, in the 1000 mg/kg group, significant increases in the absolute and relative weights of the adrenals were noted, along with a significant increase in the absolute weight of the liver, and there was an increasing trend exhibited for the absolute weights of the liver. In the females, significant increases in the absolute and relative weights of the liver were noted in the 1000 mg/kg group.

GROSS PATHOLOGY, HISTOPATHOLOGY: NON-NEOPLASTIC
Changes thought to be caused by administration of the test substance were noted in the liver, kidney and adrenals.
Of the impregnating males, three out of six in the 1000 mg/kg group exhibited central lobular hepatocellular hypertrophy. Increased deposition of hyaline droplets in the proximal tubular epithelium of the kidney was noted in six out of eight in the 100 mg/kg group, nine out of nine in the 300 mg/kg group and five in the 1000 mg/kg group, exhibiting a strong dose dependency. Additionally, there were five animals with increased basophilic tubules in the kidneys in the 300 mg/kg group, three in the 1000 mg/kg group and dilation of the distal tubules noted in three animals in the 1000 mg/kg group. Vacuolization of the cells of the zona fasciculata in the adrenals was noted in one in the 300 mg/kg group and three in the 1000 mg/kg group. Central lobular hepatocellular hypertrophy was noted in four out of four in the 1000 mg/kg group. The changes of dilation of distal tubules and fatty degeneration of the proximal tubule epithelium in the kidneys were noted in two of nine in the 300 mg/kg group and one in the 1000 mg/kg group. Vacuolization of the cells of the zona fasciculata in the adrenals was noted in one of the 300 mg/kg group. Of the males not causing successful pregnancies, two out of four in the 1000 mg/kg group exhibited central lobular hepatocellular hypertrophy, one out of one in the 300 mg/kg group and four of four in the 1000 mg/kg group exhibited increased deposition of hyaline droplets in the proximal tubular epithelium of the kidney, one of the 300 mg/kg group and three of the 1000mg/kg exhibited obvious basophilic tubules in the kidneys, one of the 1000 mg/kg group exhibited dilation of the distal tubules, two in the 1000 mg/kg group exhibited vacuolization of the cells of the zona fasciculata in the adrenals, and one exhibited hypertrophy of the zona fasciculata. Of the females not impregnated, one each in the 1000 mg/kg group exhibited central lobular hepatocellular hypertrophy and dilation of the distal tubules, and three in the 1000 mg/kg group exhibited vacuolization of the cells of the zona fasciculata in the adrenals. Central lobular hepatocellular hypertrophy was noted in the one female in the 1000 mg/kg group where all of the pups died during the lactation period. However, in males and females unsuccessful at impregnation and in females where all of the pups died, there were no changes noted in the pituitary gland and reproductive organs that were thought to be caused by administration of the test substance. In one euthanized female in the 1000 mg/kg group, the fatty degeneration of the proximal tubule epithelium and dilation of distal tubules in the kidneys noted was stronger than the changes in the survivors. Furthermore, central lobular hepatocellular hypertrophic necrosis was noted, along with necrosis of the mucous membranes of the glandular stomach and ileum, hypertrophy of the cells of the zona fasciculata in the adrenals and atrophy of the spleen and thymus. In addition to the findings given above, changes in each of the organs were noted but they were sporadic and not confirmed to be related to administration of the test substance.

Effect levels

open allclose all
Dose descriptor:
NOAEL
Effect level:
30 mg/kg bw/day (actual dose received)
Sex:
male
Basis for effect level:
other: No significant effects at this dose
Dose descriptor:
NOAEL
Effect level:
100 mg/kg bw/day (actual dose received)
Sex:
female
Basis for effect level:
other: No significant effects at this dose

Target system / organ toxicity

Critical effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
Based on the above mentioned results, the non-observable adverse effect levels for repeat dose toxicity of diacetone alcohol in rats are considered to be 30 mg/kg/day in males and 100 mg/kg/day in females.
Executive summary:

A repeated dose study according to OECD TG 422 was conducted with 4-hydroxy-4-methyl-2-pentanone (diacetone alcohol, or HMP). In this study, 10 rats/sex/group were dosed by gavage with 0, 30, 100, 300, or 1000 mg/kg/day of 4-hydroxy-4-methyl-2-pentanone in distilled water for approximately 45 days. Decreased locomotor activity and stimulation response were observed at 300 and 1000 mg/kg/day for both sexes. Altered blood parameters, dilatation of the distal tubules of the kidneys, hepatocellular hypertrophy, and vacuolization of the zona fasciculata of the adrenals were observed in males from the 1000 mg/kg/day group. Females at 300 mg/kg/day showed dilatation of the distal tubules and fatty degeneration of the proximal tubular epithelium in the kidneys. At 1000 mg/kg/day, female body weight gain was reduced, liver weight was increased, hepatocellular hypertrophy was noted, and kidney lesions similar to those at 300 mg/kg/day were also recorded. In addition, increased incidence and/or severity of hyaline droplets in the tubular epithelium was noted in males at 100 mg/kg/day and greater. The LOAEL for this study was 100 mg/kg/day for males and 300 mg/kg/day for females. The NOAEL was 30 mg/kg/day for males (based on hyaline droplet nephropathy) and 100 mg/kg/day for females.