Resin acids and Rosin acids, reaction products with formaldehyde, calcium salts

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

2007

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study is valid without restriction. According to ECHA guidance, a default of 2 is assigned because of read-across.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2007

Report date:

2007

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

version of July 27, 1995

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Dehydrogenated/partially hydrogented gum rosin

IUPAC Name:

Dehydrogenated/partially hydrogented gum rosin

Constituent 2

Reference substance name:

1740-19-8 (dehydr.) and 65997-06-0 (partially hydro.)

IUPAC Name:

1740-19-8 (dehydr.) and 65997-06-0 (partially hydro.)

Details on test material:

- Name of test material (as cited in study report): Dehydrogenated/partially hydrogented gum rosin
- Molecular formula (if other than submission substance):C20H28O2
- Physical state:solid (light yellow to amber)
- Analytical purity: not applicable, substance is UVCB
- Purity test date: Date of Certificate of Analysis not provided in report.
- Lot/batch No.: 6013
- Expiration date of the lot/batch: 16 May 2007
- Stability under test conditions: Stable
- Storage condition of test material: Ambient temperature, in the dark.
- Other: Melting point 90°C, density 1.06 g/cm3

Test animals

Species:

rat

Strain:

Fischer 344

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland GmbH
- Age at study initiation: 7 weeks
- Weight at study initiation: 183g (males) and 125 g (females) with a 20% standard deviation
- Fasting period before study: none
- Housing: 2 or 3 animals of the same sex per cage
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 5 days

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

IN-LIFE DATES: From: July 3 2006 To:November 15 2006

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: 0.1% aqueous solution of Na-carboxymethyl cellulose plus 0.01% Polyoxyethylensorbatanmonooleat

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:
The vehicle was prepared freshly in biweekly intervals and stored in the refrigerator. The test item formulation was prepared daily prior to dosing.

VEHICLE
- Justification for use and choice of vehicle (if other than water): Test item is poorly soluble. The vehicle helped forming a stable suspension.
- Concentration in vehicle: adjusted on body weight
- Amount of vehicle (if gavage): 10 ml/kg bw

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Determination of stability and homogenity were verified analytically with samples taken on day 8. An HPLC method was used.

Duration of treatment / exposure:

28 days

Frequency of treatment:

daily

Doses / concentrations

Remarks:

Doses / Concentrations:
100, 316 and 1000 mg/kg bw
Basis:
actual ingested

No. of animals per sex per dose:

5
(5 additional animals for doses with recovery groups)

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Range-finder study

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily


DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: once a week

BODY WEIGHT: Yes
- Time schedule for examinations:once a week

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: Yes

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Day 29 and Day 45
- Anaesthetic used for blood collection: Yes (ether)
- Animals fasted: Yes
- How many animals: all
- Parameters examined: Red blood cell count, haemoglobin concentration, haematocrit, mean corpuscular haemoglobin and its concentration, white blood cell count, mean cell volume, platelet count, differential white blood cell count, prothrombin time (Quick)

CLINICAL CHEMISTRY: Yes
- Animals fasted: Yes
- How many animals: all
- Parameters examined: Alanin aminotransferase, albumin, alkaline phosphatase, aspartate aminotransferase, cholesterol, creatinine, gamma-glutamyltransferase, glucose, potassium, sodium, total protein, urea

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No

Sacrifice and pathology:

GROSS PATHOLOGY: Yes

Organ weights: adrenal glands, brain, epididymides, heart, liver, kidney, spleen, testes, thymus

HISTOPATHOLOGY: Yes (all gross lesions, adrenal glands, brain, colon, duodenum, heart, iluem , jejunum, kidneys, liver, lungs, lymph nodes (mandibular, mesenteric), ovaries, prostate, sciatic nerve, spinal cord, spleen, sternum with bone marrow, stomach, testes, thymus, thyroid and parathyroid glands, trachea, urinary bladder, uterus)

Statistics:

Analysis of variance follwed by the Schffe-test, t-test, H-test of Krskal and Wallis followed by the test of Nemenyi, Chi-square test and Fisher`s exact test

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food efficiency:

no effects observed

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Description (incidence and severity):

A significant increase in creatinine and sodium was observed in high dosed animals, but the values were within the historical control range. The increase was not observed in animals of the recovery group.

Urinalysis findings:

not examined

Behaviour (functional findings):

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

centrilobular cytoplasmatic vacuolization of hepatocytes of females, still present after the 14-day recovery sacrifice.

Details on results:

None of the slight differences in the values for haematology, clinical chemistry and organ weights showed a dose-response relationship and all values were within the range of historical control values.

The test substance was found to be sufficiently stable in preparations for at least 4h. Concentrations and homogenity checks showed that these parameters were within acceptable limits.

Effect levels

1

Dose descriptor:

NOAEL

Effect level:

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

Based on:

test mat.

Sex:

female

Basis for effect level:

other: centrilobular cytoplasmatic vacuolization in hepatocytes, not reversible within 14 days.

2

Dose descriptor:

NOAEL

Effect level:

>= 1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male

Target system / organ toxicity

Critical effects observed:

not specified

Any other information on results incl. tables

mean clinical biochemistry data, males
parameter	control	100 mg/kg bw	316 mg/kg bw	1000 mg/kg bw	recovery control	recovery high dose
Cholesterol	1.47	1.49	1.48	1.52	1.44	1.51
AST	76.2	82.4	81.2	85.6	79.8	87.8
ALT	60.2	63	56.4	52.8	55.6	60.4
GGT	0.58	0.46	0.26	0.38	0.5	0.44
AP	183	153	202	204	209	166
UREA	7.93	7.93	8.31	7.11	8.67	9.36
CREA	30.4	30.8	30.6	32.4	35.4	32.7
GLU	6.43	6.75	5.55	5.91	6.11	6.81
TP	70	71.5	71.2	71.4	72.8	73.4
ALB	43.6	42.4	43.7	44.1	45.4	43.6
Na	140	141	140	140	141	140
K	3.3	3.3	3.32	3.3	3.16	3.18
mean clinical biochemistry data, females
parameter	control	100 mg/kg bw	316 mg/kg bw	1000 mg/kg bw	recovery control	recovery high dose
Cholesterol	1.92	1.97	1.92	1.93	1.92	1.82
AST	88.5	84.8	85.4	89.2	93	85.8
ALT	52.2	57.4	50.4	51.4	60	51.8
GGT	0.6	0.74	0.46	0.6	0.44	0.74
AP	177	142	178	172	176	157
UREA	7.31	7.7	7.5	7.73	8.44	7.13
CREA	26.7	28.8	30.1	29	33.9	29.5
GLU	5.01	5.02	4.36	4.41	4.39	5
TP	67	70.4	67.1	68	69.1	67.7
ALB	43.4	43.1	43.2	44.5	44.6	42.2
Na	138	140	139	140	140	140
K	3.44	3.3	3.32	3.2	3.22	3.14

Details on the histopathology findings in liver:

One female of the control/recovery group showed minimal cytoplasmatic centrilobular vacuolization of hepatocyes.

Both at the end of dosing and after recovery, in the high dosed females the same was observed with minimal and mild severety in one and three animals, respectively.

No histopathology findings were observed in males.

In the high dosed recovery group females,

Applicant's summary and conclusion

Conclusions:

No adverse effects upon subacute oral gavage dosing if rats was observed at 316 mg/kg bw.