Registration Dossier

Administrative data

Endpoint:
sub-chronic toxicity: oral
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
1986-12-02 to 1987-03-06
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: This study is classified as reliable without because it was a well conducted study.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1990
Report Date:
1990

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents)
Deviations:
yes
GLP compliance:
not specified
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Test material form:
other: Oily liquid
Details on test material:
Read across from untreated distillate aromatic extracts

- Name of test material (as cited in study report): 318 Isthmus Furfural Extract
- Substance type:heavy paraffinic distillate solvent extract (petroleum)
- Physical state: liquid
- Analytical purity: not provided
- Impurities (identity and concentrations): not provided
- Composition of test material, percentage of components (wt. %): total non-aromatics- 22.3, total aromatics- 77.7%, less than 3 ring polynuclear aromatic hydrocarbons- 37.2%, 3 to 5 ring polynuclear aromatic hydrocarbons- 23.0%, N-polynuclear aromatic hydrocarbons- 2.3% (non-basic = 1.6; basic 0.7), S-polynuclear aromatic hydrocarbons- 12.8%
- Isomers composition: not provided
- Purity test date: not provided
- Lot/batch No.: CRU no. 86187
- Expiration date of the lot/batch: April 30, 1991
- Stability under test conditions: 5 years
- Storage condition of test material: not provided

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Taconic, Germantown, NY
- Age at study initiation: 49 days
- Weight at study initiation: not reported
- Fasting period before study: not reported
- Housing: individual housing in hanging stainless steel cages with wire bottoms and fronts
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 13 days in a quarantine room


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 to 21°C
- Humidity (%): 40 to 60%
- Air changes (per hr): not reported
- Photoperiod (hrs dark / hrs light): 12 hours light/12 hours dark

IN-LIFE DATES:
From 1986-12-02 to 1987-03-06

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
not specified
Details on oral exposure:
No data provided.
Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
No data reported.
Duration of treatment / exposure:
13 weeks
Frequency of treatment:
Five days per week
Doses / concentrations
Remarks:
Doses / Concentrations:
125 or 500 mg/kg/day via oral gavage
Basis:
no data
No. of animals per sex per dose:
10 male rats per dose
Control animals:
yes, concurrent no treatment
Details on study design:
No data reported.
Positive control:
No data reported.

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice a day
- Cage side observations included morbidity and mortality.


DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily


DERMAL IRRITATION (if dermal study): Yes
- Time schedule for examinations: weekly


BODY WEIGHT: Yes
- Time schedule for examinations: study initiation and weekly


FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No


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: No


WATER CONSUMPTION: No


OPHTHALMOSCOPIC EXAMINATION: No


HAEMATOLOGY: Yes
- Time schedule for collection of blood: weeks 5 and 13
- Anaesthetic used for blood collection: Yes (diethyl ether)
- Animals fasted: Yes
- How many animals: all animals
- Parameters in table 1 were examined.


CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: weeks 5 and 13
- Animals fasted: Yes
- How many animals: all animals
- Parameters in table 2 were examined.


URINALYSIS: Yes
- Time schedule for collection of urine: weeks 5 and 13
- Metabolism cages used for collection of urine: No data
- Animals fasted: Yes
- Parameters in table 3 were examined.


NEUROBEHAVIOURAL EXAMINATION: No

Sacrifice and pathology:
GROSS PATHOLOGY: Yes, but details were not provided.
HISTOPATHOLOGY: Yes (see table 4)
Other examinations:
Organ weights double XX in table 4 were weighed. Sperm morphology was examined in the 500 mg/kg/day group.
Statistics:
An analysis of variance with associated F-test followed by Student-Newman-Keuls test were used when appropriate.

Results and discussion

Results of examinations

Clinical signs:
effects observed, treatment-related
Mortality:
mortality observed, treatment-related
Body weight and weight changes:
effects observed, treatment-related
Food consumption and compound intake (if feeding study):
not examined
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
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:
not examined
Details on results:
CLINICAL SIGNS AND MORTALITY: Four of ten mice in the 500 mg/kg/day group were sacrificed prior to scheduled termination. All animals in the 125 mg/kg/day survived to date of sacrifice. No details on clinical signs were provided.


BODY WEIGHT AND WEIGHT GAIN: Body weight was significantly reduced in the 500-mg/kg/day group (Table 1)



HAEMATOLOGY:A significant decrease (p<0.05) in red blood cell (RBC) parameters (including RBC count, haemoglobin, and haematocrit) and platelet in males dosed orally at 500 mg/kg/day(Table 2). Males orally dosed at 125 mg/kg/day showed a significant decrease in RBC parameters; platelet counts were slightly decreased in these rats but did not achieve statistical significance. There were no significant differences in the RBC morphology or WBC differential data.


CLINICAL CHEMISTRY: The only statistically significant difference between the serum data from control and orally dosed rats was observed for SDH (0 mg/kg/day = 5±2 IU/l, 150 mg/kg/day = 8±2 IU/l, 500 mg/kg/day = 9±7 IU/l).


URINALYSIS: There were no treatment-related effects on urinalysis.


ORGAN WEIGHTS:Significantly different absolute and/or relative weights in liver, heart, brain, thymus, prostate, seminal vesicles and adrenal glands compared to control were observed. Some of the effects are secondary to reduced body weight. Those that are considered related to treatment are presented in Table 3.

GROSS PATHOLOGY: Focal areas of red discoloration and or generalized reddening were also observed in the brain, spinal cord, stomach and testes of many of the rats dosed orally at 500 mg/kg/day.


HISTOPATHOLOGY: Treatment-related histopathology was generally dose-dependent and occurred in the following tissues: adrenals, bone marrow, liver, stomach and thymus (Table 9). Atrophy occurred in the male sex organs (testes, seminal vesicle, and prostate).


OTHER FINDINGS: Sperm evaluations showed a significant increase in the frequency of sperm with abnormal heads in the rats dosed orally at 500 mg/kg/day (1.9% in controls and 3.2% in treated rats).

Effect levels

open allclose all
Dose descriptor:
NOAEL
Sex:
male
Basis for effect level:
other: adverse effects observed at 125 mg/kg/day dosed orally
Remarks on result:
not determinable
Remarks:
no NOAEL identified
Dose descriptor:
LOAEL
Effect level:
125 mg/kg bw/day (nominal)
Sex:
male
Basis for effect level:
other: overall effects

Target system / organ toxicity

Critical effects observed:
not specified

Any other information on results incl. tables

Table 1. Mean group body weights on week of study.

Dose Week 1 Week 2 Week 3 Week 4 Week 5 Week 6 Week 7 Week 8 Week 9 Week 10 Week 11 Week 12 Week 13 Week 14
0 mg/kg/day 227.2 273.9 318.6 348.5 - 380 405.4 419.1 433.2 440.4 462.4 460.8 468.9 461.5
125 mg/kg/day 224.4 270.9 314.5 347.7 - 375.4 395 403.9 416 422.7 431.6 439 451.8 445.5
500 mg/kg/day 224.1 285.3 308.9 335.3 - 348.8* 354.3* 352.2* 359.9* 359.1* 373.2* 374.6* 380.1* 386.5*

*indicates significantly different from control (p<0.05)

Body weights were inadvertently not measured on week 5.

Table 2. Haematological parameters at the end of the study period.

Haematological Parameter 0 mg/kg/day 125 mg/kg/day 500 mg/kg/day
RBC(106/mm3) 9.17 8.72 8.51*
WBC (103/mm3) 14.4 13.3 12.7
Platelets (10^3/mm3) 1152 1055 930
Hgb (g/dl) 17.1 16.0* 15.2*
Hct (%) 53.7 50.7* 47.7*
MCV (cubic microns) 59 58 56*
MCH (pcg) 18.7 18.4 17.8* 
MCH Conc. (%) 31.9 31.6 31.8

*indicates significantly different from control (p<0.05)

Abbreviations: RBC=red blood cells, WBC=white blood cells, Hgb=haemoglobin, Hct=haematocrit,

MCV=mean corpuscular volume, MCH=mean corpuscular haemoglobin

 

Table 3. Mean final fasted body weight and relative organ weights in grams.

Body/Organ Weight

0 mg/kg/day

125 mg/kg/day

500 mg/kg/day

Final Body Weights

435.1±36.7

407.1±32.6

350.3±29.4*

Liver

3.03±0.24

4.17±0.40*

5.29±0.46*

Prostate

0.26±0.03

0.20±0.05*

0.11±0.07*

Seminal Vesicles

0.20±0.04

0.18±0.05

0.12±0.04*

Thymus

0.093±0.029

0.054±0.015*

0.018±0.007*

 

Table 4

 Incidences of treatment-related histopathology after dermal exposure to 318 Isthmus Furfural Extract for 13 weeks

 

Control

125 mg/kg/day

500 mg/kg/day

Males

n=10

n=10

n=10

vacuolation, cortical

0

 

 

Not examined

3

hyperplasia, zona glomerulosa

 

0

Not examined

2

decreased cellularity in femur

0

4

8

Fibrosis in femur

0

1

5

decreased cellularity in sternum

0

2

7

Fibrosis in sternum

0

0

4

haemorrhage, multifocal (brain)

0

Not examined

5

Dialation, sinusoidal, centrilobular

0

0

3

Hypertrophy, hepatocytes

0

0

9

Necrosis, centrilobular

0

0

4

Vacuolation, hepatocellular, periportal

0

2

5

Atrophy (prostate)

0

Not examined

9

Atrophy (seminal vesicle)

0

Not examined

5

Congestion/haemorrhage, glandular mucosa

0

Not examined

4

Hyperplasia/hyperkeratosis, limiting ridge

0

Not examined

8

Atrophy (thymus)

0

9

10

 

Applicant's summary and conclusion

Conclusions:
NOAEL for heavy paraffinic distillate aromatic extract could not be identified and is less than 125 mg/kg/day when administered orally.
Executive summary:

Read across justification

This compound is an untreated distillate aromatic extract and provides a worst case scenario for insufficiently refined other lubricant base oils due to the concentration effect of the solvent extraction process.

In a subchronic oral toxicity study, heavy paraffinic distillate aromatic extract was administered to 10 male Sprague-Dawley rats/dose at dose levels 0, 125, or 500 mg/kg bw/day 5 days a week for 13 weeks. Four of ten mice in the 500 mg/kg/day group were sacrificed prior to scheduled termination. All animals in the 125 mg/kg/day survived to date of sacrifice. No details on clinical signs were provided. Body weight was significantly reduced in the 500-mg/kg/day group. A significant decrease (p<0.05) in red blood cell (RBC) parameters (including RBC count, haemoglobin, and haematocrit) and platelet in males dosed orally at 500 mg/kg/day. Males orally dosed at 125 mg/kg/day showed a significant decrease in RBC parameters; platelet counts were slightly decreased in these rats but did not achieve statistical significance. There were no significant differences in the RBC morphology or WBC differential data. The only statistically significant difference between the serum data from control and orally dosed rats was observed for SDH (0 mg/kg/day = 5±2 IU/l, 150 mg/kg/day = 8±2 IU/l, 500 mg/kg/day = 9±7 IU/l). Treatment-related dose-dependent changes in relative organ weights included increased liver weight in both groups, decreased prostate weight in both groups, decreased seminal vesicle weight in the high-dose group, and decreased thymus weight in both groups. Focal areas of red discoloration and or generalized reddening were also observed in the brain, spinal cord, stomach and testes of many of the rats dosed orally at 500 mg/kg/day. Treatment-related histopathology was generally dose-dependent and occurred in the following tissues: adrenals, bone marrow, liver, stomach and thymus. Atrophy occurred in the male sex organs (testes, seminal vesicle, and prostate). Sperm evaluations showed a significant increase in the frequency of sperm with abnormal heads in the rats dosed orally at 500 mg/kg/day (1.9% in controls and 3.2% in treated rats). 

NOAEL for heavy paraffinic distillate aromatic extract could not be identified and is less than 125 mg/kg/day when administered orally.

This study received a Klimisch score of 1 and is classified as reliable without restriction because it is a well conducted study.