Reference type

Author

Year

Title

Bibliographic source

Testing laboratory

Report no.

Owner company

Company study no.

Report date

study report

MHLW Japan

2009

Combined repeat dose and reproductive/developmental toxicity screening test of 2,3,4,4'-Tetrahydroxybenzophenone by oral administration in rats

available in the web of Japan Existing Chemical Data Base (JECDB) at http://dra4.nihs.go.jp/mhlw_data/jsp/SearchPageENG.jsp

BoZo Research Center

data published

7.8.1 Reproductive/developmental toxicity.001

combined repeated dose and reproduction / developmental screening

no

Qualifier

Guideline

Deviations

according to

OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)

no

yes

yes

Identifier

Identity

CAS number

31127-54-5

- Name of test material (as cited in study report): 2,3,4,4'-Tetrahydroxybenzophenone
- Purity: 99.91%
- Impurities (identity and concentrations): Unknown
- Lot/batch No.: JSCXB
- Stability under test conditions: Stable
- Storage condition of test material: Refrigeration
- Dosing solution storage condition: Room temperature and protected from light
- Other: The dosing solution was used within 7 days of preparation.

rat

other: Crl: CD(SD)

male/female

TEST ANIMALS
- Source: Charles River Laboratories Japan, Inc. Atsugi
- Age at study initiation: 10 weeks
- Weight at study initiation: Males: 335-391 g; Females: 202-249 g
- Housing: Steel wire-mesh cage (250 mm x 350 mm x 200 mm )
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 14 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21-23
- Humidity (%): 46-61
- Air changes: 10-15 times / hr
- Photoperiod: 12 hrs dark / 12 hrs light

oral: gavage

corn oil

PREPARATION OF DOSING SOLUTIONS:
VEHICLE
- Amount of vehicle (if gavage): 5 mL/kg bw
- Lot/batch no. (if required): SDE2487

(P) Males: 42 days including 14 days pre-mating and mating periods, and thereafter 14 days (P) Females: 42-55 days including 14 days pre-mating, mating and gestation periods, and the days until day 4 of lactation; satellite animals: 42 days.

Once/day, 7days/week

0 (vehicle), 100, 300, and 1000 mg/kg bw/day

Basis

actual ingested

12 animals/sex/dose as a main dose group,
5 males and 5 females at 0 and 1000 mg/kg bw/day as a satellite group (without mating)

yes, concurrent vehicle

- Dose selection rationale: Doses in this test were set based on the results of the following study: 14-day repeated dose oral toxicity test (doses: 100, 300, and 1000 mg/kg bw/day). In the 14-day repeated dose oral toxicity test, abnormalities were observed in animals in the 1000 mg/kg bw/day group, such as low values of body weight and food consumption an increase in liver weight. No effects were observed at 300 mg/kg bw/day. On the basis of these effects, a dose level of 1000 mg/kg was selected as the maximum dose expecting to induce the toxic changes, and then dose levels of 300 and 100 mg/kg bw/day were selected as a middle dose and a minimum dose levels, respectively, in accordance with a common ratio of approximately 3.
- Rationale for animal assignment (if not random): Body weight-balanced randomization
- Post-exposure recovery period in satellite groups: 14 days

CAGE SIDE OBSERVATIONS: Yes
- Time schedule:
Males and females: once before the start of administration, 3 times/day during the administration period, and once during the recovery period

DETAILED CLINICAL OBSERVATIONS: Yes
The functional observational battery testing (FOB) was performed on all animals. Among the measures in the FOB, detailed clinical observations were made before the initiation of dosing. Thereafter, in males of the main groups, detailed clinical observations were made once a week. Also in females of the main groups, detailed clinical observations were made once a week in pre-mating and mating periods thereafter, and then those were made on days 1,7,14 and 20 of gestation, and on day 4 of lactation. For the satellite group, detailed clinical observations were made once a week in dosing and recovery periods.
Sensory motor reflexes, forelimb and hindlimb grip strengths, and motor activity were measured on week 6 of administration period (main/recovery group animals) and week 2 of recovery period (recovery group animals).

BODY WEIGHT: Yes
- Time schedule for examinations: Males (main) & males and females (recovery group): Days 1, 4, 8, 11, 15, 22, 25, 29, 32, 36, 39, 42, and the day of necropsy (after ca. 16h-fasting) in dosing period
Males and females (recovery group): Days 1, 4, 8, 11, 14, and the day of necropsy (after ca. 16h-fasting) in recovery period
Females (main group): Twice a week during the precopulation period (days 1, 4, 8, 11, and 15); gestation days 0, 4, 7, 11, 14, 17, and 20; lactation days 0 and 4; and the day of necropsy (after ca. 16 h-fasting)

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption: Yes
Males (main) & males and females (recovery group): Days 1, 4, 8, 11, 15, 32, 36, and 39 in dosing period
Males and females (recovery group): Days 1, 4, 8, 11, and 14 in recovery period
Females (main group): Days 1, 4, 8, 11, and 15; gestation days 1, 4, 7, 11, 14, 17, and 20; lactation days 2 and 4

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Blood was collected on the day of necropsy
- Anaesthetic used for blood collection: Yes (ether)
- Animals fasted: Yes, 16-20h
- How many animals: 5 sex/dose/group
- Parameters checked in table were examined.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Same as hematology
- Animals fasted: Same as hematology
- How many animals: Same as hematology
- Parameters checked in table were examined.

URINALYSIS: Yes (males only)
- Time schedule for collection of urine: Day 36-37 in dosing period, day 8-9 in recovery period
- Metabolism cages used for collection of urine: No data
- Animals fasted: fasting and only water at libitum (4h-urine), no fasting (20h-urine)

GROSS PATHOLOGY: Yes, whole organs and tissues (see tables)
HISTOPATHOLOGY: Yes (see tables)

Organ weight: Brian, thyroids (including parathyroids), thymus, heart, liver, spleen, kidneys, adrenals, testes, epididymis

The data were analyzed for homogeneity of variance by the Bartlett test. If variances were homogeneous, data was analyzed by the Dunnett test, whereas heterogeneous data was analyzed by the Dunnett type mean rank test (p<0.05, two-sided).
In the recovery test, these values of two groups were analyzed by F test. If variances were homogeneous, data was analyzed by the Student t-test, whereas heterogeneous data was analyzed by the Aspin-Welch t-test (p<0.05, two-sided).

Endpoint

Effect level

Based on

Sex

Basis for effect level / Remarks

LOAEL

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

test mat.

male/female

effects on the cecum

yes

yes

yes

not examined

not examined

not examined

yes

yes

yes

yes (see clinical signs.)

yes

yes

yes

not examined

CLINICAL SIGNS AND MORTALITY
Males: No dead or moribund animals were observed. Salivation was observed at 1000 mg/kg bw/day.
Females: After delivery (day 0 of lactation), one animal died at 1000 mg/kg bw/day.
Home cage observation: No effects.
In-the-hand observation: No effects.
Open field observation: No effects.
-Sensory motor reflexes:
Auditory response, approach response, touch response, tail pinch response, pupillary reflex, aerial righting reflex: No effects. Landing foot splay: High value in females at 1000 mg/kg bw/day (main group, day 4 of lactation)
-Forelimb and hindlimb grip strengths: Low value of hindlimb grip strength in males at 1000 mg/kg bw/day in the 6-week of dosing
-Motor activity: High value in females at 300 mg/kg bw/day (main group, day 4 of lactation) without dose-response relationship. In females, high value at 1000 mg/kg bw/day in the 6-week of dosing in the recovery group, but not in the main group at the same dose. (No toxicological effects)

BODY WEIGHT AND WEIGHT GAIN
Males: Low values of body weight gain was observed at 1000 mg/kg bw/day in main group.
Females: Low value of body weight gain was observed at 1000 mg/kg bw/day at the ends of pre-mating and gestation periods. Low value of body weight was observed at 300 mg/kg bw/day on day 4 of lactation.
At the end of recovery period, high value of body weight gain was observed in both sexes at 1000 mg/kg bw/day.

FOOD CONSUMPTION
Males: Low value of food consumption was observed at 1000 mg/kg bw/day on day 4 of dosing, and significant increase was observed on days 36-42.
Females: Low value of food consumption was observed at 1000 mg/kg bw/day on day 4 of dosing, high value on day 15 of dosing, and then low value on day 20 of gestation and day 2 of lactation. Low value of food consumption was observed at 300 mg/kg bw/day on day 4 of dosing and day 2 of lactation.

HAEMATOLOGY
Low values of RBC, Hb, Ht and MCHC, and high values of platelet, neutrophilic count and monocyte count were observed at 1000 mg/kg bw/day at the end of dosing. At the end of recovery period, low values of RBC and Hb, and high value of reticulocyte were observed in males at 1000 mg/kg bw/day. Other significant findings in the table were within the normal ranges of physiological variability.

CLINICAL CHEMISTRY
High value of inorganic phosphorus was observed at 300 mg/kg bw/day and more at the end of dosing. Other significant findings in the table were within the normal ranges of physiological variability.

URINALYSIS (only males; statistical analysis was not performed on qualitative items.)
Occult blood was observed in all administered males, and the degree of occult blood was enhanced in a dose-dependent manner. Dark yarrow color was observed in each 2-3 males at 100 mg/kg bw/day and more.

ORGAN WEIGHTS
Low values of absolute and relative thymus weights were observed in females at 300 mg/kg bw/day and more at the end of dosing. High value of relative liver weight was observed in both sexes at 1000 mg/kg bw/day at the end of dosing, and in females at the end of recovery period. Other significant findings in the table were within the normal ranges of physiological variability.

GROSS PATHOLOGY
In the dead female at 1000 mg/kg bw/day, small sizes of spleen and thymus were observed.
Undernourishment of general descriptions was observed in one female at 1000 mg/kg bw/day. Dark discoloration of liver was observed in six males at 1000 mg/kg bw/day. Small size of thymus was observed in one female at 300 mg/kg bw/day, and in three females at 1000 mg/kg bw/day. Other findings in the table were considered to be incidental due to low frequency of appearance and/or pathological properties.

HISTOPATHOLOGY: NON-NEOPLASTIC
In the dead female at 1000 mg/kg bw/day, atrophy of white pup in the spleen and atrophy of thymus were observed.
[At the end of dosing]
Cecum: Single cell necrosis of mucosal epithelial cells was observed in 4 males and 2 females at 100 mg/kg bw/day, in 3 males and 3 females at 300 mg/kg bw/day, and in 8 males and 7 females at 1000 mg/kg bw/day. Diffuse hyperplasia of mucosa was observed in 1 male and 1 female at 100 mg/kg bw/day, in 3 males and 4 females at 300 mg/kg bw/day, and in 7 males and 6 females at 1000 mg/kg bw/day.
Liver: Vacuolation of peripheral hepatocytes was dose-dependently decreased in males and females at 300 mg/kg bw/day and more.
Thymus: Atrophy was dose-dependently increased in females at 300 mg/kg bw/day and more.
[At the end of recovery period]
Cecum: Diffuse hyperplasia of mucosa was observed in one male at 1000 mg/kg bw/day.
Other findings in the tables were considered to be incidental due to low frequency of appearance and/or pathological properties.

Based on the effects on the cecum, the low observed adverse effect level (LOAEL) for repeated oral dosing was determined to be 100 mg/kg bw/day in male and female rats.

UUID

IUC5-9c9461e4-d1ec-4fcd-aa97-faf2ecde802a

Dossier UUID

0

Author

dra / National Institute of Health Sciences / Tokyo / Japan

Date

2016-12-21 14:50:21 JST

Remarks

Purpose flag

key study

Study result type

experimental result

Reliability

1 (reliable without restriction)

Rationale for reliability incl. deficiencies

OECD Test Guideline study under GLP condition