Reaction product of fatty acids C14-18/C16-18 unsaturated with dihydrogendioxide and Ammonia

Repeated dose toxicity: oral

Administrative data

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study (OECD).

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1995

Report date:

1995

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)

Version / remarks:

September 21, 1998

Deviations:

yes

Remarks:

no observations for neurological endpoints

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Reaction product of fatty acids C14-18/C16-18 unsaturated with dihydrogendioxide and Ammonia

Molecular formula:

Reaction product of fatty acids C14-18/C16-18 unsaturated with dihydrogendioxide and Ammonia

IUPAC Name:

Reaction product of fatty acids C14-18/C16-18 unsaturated with dihydrogendioxide and Ammonia

Details on test material:

-Name of test material (as cited in study report): DHW 80
- Physical state: liquid up to pasty
- colour: light brown
- Composition of test material, percentage of components: active substance content: 49 - 51 %
- Lot/batch No.: 1043/3
- Expiration date of the lot/batch: December 1994
- Stability under test conditions: stable in aqua dest.
- Storage condition of test material: at room temperature
- Other:
SIS-No.: 9032
RIS-No.: 51Z6300295000
SAT-No.: 930935

Test animals

Species:

rat

Strain:

other: Hsd/Win:Wu

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Winkelmann, D-33178 Borchen
- Age at study initiation: approximately 5 weeks
- Weight at study initiation: 49g - 63g for males; 49g - 61g for females
- Housing: three or two animals in Makrolon Type M 5 cages (E. Becker & Co. GmbH, D-44579 Castrop-Rauxel) with standard softwood bedding (ARWI-Center, D-45307 Essen)
- Diet: Pelleted Altromin Maintenance Diet 1324 (Altromin GmbH, D-32770 Lage), ad libitum
- Water: Community Tap Water from Düsseldorf ad libitum (anlaysed monthly for use as drinking water)
- Acclimation period: 7 days under laboratory conditions

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 24°C
- Humidity (%): 39 - 64 %
- Air changes (per hr): 10 - 15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescent light/12 hours dark

IN-LIFE DATES: From: 1994-01-24 To: 1994-04-28/29 and recovery groups 1994-5-27

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:
- daily before administration
- administration was carried out within approximately 2-3 hours

VEHICLE
- Concentration in vehicle:
0.6% = 0.26% (active content = 49 - 51%)
1.5% = 0.67% ( ~ )
3.0% = 1.3% ( ~ )
10.0% = 4.6% ( ~ )
- Amount of vehicle (if gavage): 10 mL/kg bw

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

- analysed by Henkel TTA-Analytical Center in the time between 1994-02-21 to 1994-02-28 (study-no.: TTA 94-1861)
- concentrations of the test substance in aqua dest. based upon the results of determination of dry sediment

Duration of treatment / exposure:

main groups: 90 days, recovery groups were observed for an additional 28 days

Frequency of treatment:

once daily on working days

Doses / concentrations

Doses / concentrations 1

Remarks:

Doses / Concentrations:
60, 150, 300, 1000 mg/kg bw
Basis:
actual ingested

Doses / concentrations 2

Remarks:

Doses / Concentrations:
0.6, 1.5, 3.0, 10.0 mg/mL
Basis:
other: concentration

No. of animals per sex per dose:

10 male and female (main study groups)
5 male and female (recovery control and high dose group)

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: acute oral toxicity limit test in rats (HenkelTBD900783) with 5 animals per sex, where LC50 value was determined to be > 2000 mg/kg bw
- main dose groups 1-5 and recovery groups see table1
- Post-exposure recovery period in satellite groups: 4 weeks

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: acclimatization period 1x daily, treatment period 2x daily, treatment-free period 1x daily, (weekend and public days 1x daily)

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: acclimatization period 1x daily, treatment period 2x daily, treatment-free period 1x daily, (weekend and public days 1x daily)

BODY WEIGHT: Yes
- Time schedule for examinations: at arrival, acclimatization time 1x, weekly during the treatment period and the treatment-free period

FOOD CONSUMPTION:
- Food consumption male/female determined and mean weekly diet consumption calculated as g/rat/week: Yes
- Mean food conversion for male/female was calculated over weeks 1-6 and 8-13.

WATER CONSUMPTION: Yes
- Time schedule for examinations: weekly, weighing per cage with the exception of the dates for clinical chemistry

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: at the beginning (by slit lamp microscope Topcon-SL 5D (Bon, D-23812 Wahlstedt)) and end (hand slit lamp (Zeiss, D-73447 Oberkochen)) of treatment, after instillation of a mydriatic agent into the eyes (Mydriaticum Stulln R; Pharma Stulln GmbH, D-92551 Stulln/Nabburg)
- Dose groups that were examined: all main dose groups

HAEMATOLOGY: Yes
- Time schedule for collection of blood: after 6 weeks of administration (intermediate analysis) and at the end of treatment (final analysis)
- How many animals: all animals
- Parameters checked in table2 were examined.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: after 6 weeks of administration (intermediate analysis) and at the end of treatment (final analysis)
- Animals fasted: no data
- How many animals: all
- Parameters checked in table2 were examined.

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No

OTHER:
- bone marrow smears (at scheduled necropsy, will be examined to a special indication)
- serum samples (as blood samples but in dry assay tubes)

Sacrifice and pathology:

GROSS PATHOLOGY: Yes (see table3)
HISTOPATHOLOGY: Yes, examinations were performed on all organs of male and female rats of groups 1 and 5; all organs/tissue samples were fixed and preserved in 10% neutral formalin (Formal-Fixx-Concentrate; Fa. Shandon, D-60437 Frankfurt/Main)

Statistics:

- t-Test for determination of significant differences between the groups for body weigth gains (L- Sachs, Statistische Auswertmethoden, 3. Auflage, S.11, 212, Springer-Verlag, Berlin, 1971)
- t-Test for determination of significant differences between the groups for hematological and clinical chemistry examinations (1. Charles W. Dunnett: A Multiple Comparison Procedure for Comparing Several Treatments with a Control, Journal of the American Statistical Association, Dec. 1995, Vol., No. 372, 1096-1121; 2. Charles W. Dunnett: New Tables for Multiple Comparison with a Control, Biometrics, 1964, 482-490)
- Steel-Test for determination of significant differences between the groups for organ weight gains (Robert G. D: Steel: A multiple Comparison Rank Sum Test: Treatments verus Control, Biometrics, 15, (1959), 560-572

Results and discussion

Results of examinations

Details on results:

CLINICAL SIGNS AND MORTALITY
After 3 weeks application males and females of group 5 (including recovery group) showed protrusion of head into the bedding, up to high-grade reddish and brownish salivation immediately after application. There were no unscheduled deaths due to treatment which were considered as substance related. One female died due to bloodletting after application 32, one male was sacrificed in extremis after application 50 due to failure of application technique (histologically assessed).

BODY WEIGHT AND WEIGHT GAIN, FOOD EFFICIENCY
no effects

WATER CONSUMPTION
The female group 5 showed an increased consumption over the total study period, however mean water conversion remained unchanged compared to control and alterations related to the increased water consumption could not be observed.

OPHTHALMOSCOPIC EXAMINATION
no effects

HAEMATOLOGY
Only females in group 4 showed a slight decrease in lymphocytes at intermediate analysis. At final analysis no differences could be observed.

CLINICAL CHEMISTRY
Slight effects could only be seen in male animals.
At intermediate analysis: Group 2 showed a slight decrease of calcium and a slight increase of bilirubin. Group 3 a slight decrease of calcium. Group 4 a decrease of calcium and slight increase of creatinine. Group 5 showed an increase of calcium.
At final analysis: Group 3 and 4 showed slight increase of the AST.
However, significant findings observed were within the normal range of the ± 1 until ± 2s deviations of the historical values and were considered within the normal range of historical values.

ORGAN WEIGHTS
For male groups 2, 3 and 5 a decrease of the brain could be observed. Female group 4 showed a slight increase of the spleen.

GROSS PATHOLOGY
no effects

HISTOPATHOLOGY: NON-NEOPLASTIC/NEOPLASTIC
Findings consisted of spontaneous lesions in the male and female animals of all groups such as hyperplasia of the mammary gland, accumulations of lymphocytes in lungs, hydrometriosis of the uterus and other spontaneous lesions. Acute and chronic lesions of the eyes due to bloodletting were observed occasionally. Macroscopical and histopathological lesions were not treatment related.
The liver of 3 treated male animals (group 5, 4, 3) showed small areas of necrosis with an acute up to chronic character. Areas of necrosis were considered incidental and not related to the substance. Animals of recovery groups were not examined since no target organ could be assessed in the main groups.

Effect levels

Dose descriptor:

NOEL

Effect level:

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

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no substance related deaths observed

Target system / organ toxicity

Critical effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

According to the study described, a daily administration of the test substance revealed no systemic or toxic effects in all test groups. Thus, the NOEL-value (NO OBSERVED EFFECT LEVEL) is to determine at 1000mg/kg body weight/day.