Registration Dossier

Administrative data

Description of key information

By analogy with 1-fluoro-4-nitrobenzene, methemoglobinemia and damages to the spleen have been observed. Oral NOAEL is 5 mg/kg and inhalation NOAEC is 13 mg/m3.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
No data
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study well conducted, sufficiently detailed, according to standardised guidelines but not GLP.
Qualifier:
no guideline followed
Principles of method if other than guideline:
The test substance is orally administered daily in graduated doses to several groups of experimental animals, one dose level per group for a period of 5 weeks. During the period of administraion the animals are observed closely, each day for signs of toxicity.
GLP compliance:
no
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
No data
Route of administration:
oral: gavage
Vehicle:
olive oil
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
5 weeks
Frequency of treatment:
daily
Remarks:
Doses / Concentrations:
5, 15 and 45 mg/kg bw/day
Basis:
actual ingested
No. of animals per sex per dose:
10 per sex per dose
Control animals:
yes, concurrent vehicle
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: No

DETAILED CLINICAL OBSERVATIONS: No

BODY WEIGHT: YES

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


OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: after 2, 5, 7 and 9 weeks
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: 5M/5F after 2 weeks, 5M/5F after 5 weeks, 5M/5F after 7 weeks and 5M/5F after 9 weeks
- Parameters checked in table [No.?] were examined.

CLINICAL CHEMISTRY: No

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes (liver, kidneys and spleen)
Other examinations:
No
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:
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:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Histopathological findings: neoplastic:
not examined
Details on results:
PRE-TEST RESULTS:
In a pre-test, rats were orally administered the test substance at 50, 100 and 200 mg/kg bw/day. At 200 mg/kg bw/day, mortality was rapidly induced by the test substance in all animals. At 100 and 50 mg/kg bw/day, hematological effects were observed : anaemia with reticulocytosis and increasedof blood platelet (methemoglobinemia: 20 %).
At gross necropsy an enlargement of the spleen was observed. Histopatology results showed a significant effect of the test substance at 100 mg/kg bw/day at liver, spleen, kidney and testicules level. At 50 mg/kg bw/day, an effect at the spleen level was only observed.

MAIN TEST RESULTS:

CLINICAL SIGNS AND MORTALITY
No mortality was observed at any tested doses.

BODY WEIGHT AND WEIGHT GAIN
The weigth gain was similar between control and treated animals.

HAEMATOLOGY
A hemaological toxicity was observed after treatment with graduated doses of the test substance. (cf. Table 1)
Effect on percentage of Methemoglobinemia, significant at 15 and 45 mg/kg/d. This effect is reversible after 2 weeks of rest

GROSS PATHOLOGY
At 45 mg/kg bw/day, all animals showed hypertrophy of the spleen.

HISTOPATHOLOGY: NON-NEOPLASTIC
After 5 weeks, histological results showed a dose-dependant congestion of the spleen with a splenic hemosiderosis. After 9 weeks, only clusters of hemosiderin in macrophages were observed. After 4 weeks of rest, the spleen congestion disappeared, but hemosiderosis is observed and important at 45 mg/kg/d.
Key result
Dose descriptor:
NOEL
Effect level:
5 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Methemoglobinemia
Key result
Critical effects observed:
yes
Lowest effective dose / conc.:
5 mg/kg bw/day (nominal)
System:
haematopoietic
Organ:
blood
spleen
Treatment related:
yes
Dose response relationship:
yes
Relevant for humans:
yes

Table 1: Hematological effect of the test substance

Treatment

Erythroctes (E+03) and

Methemoglobinemia (%)

2 weeks 5 weeks 7 weeks 9 weeks
Control 7840 8495
(3.32)
8196
(2.0)
8002
(2.2)
45 mg/kg bw/day 6050 6536
(9.30)
7662
(3.5)
7828
(3.0)
15 mg/kg bw/day 6454 7277
(8.76)
/ /
5 mg/kg bw/day 6516 7940
(5.70)
/ /
Conclusions:
Under the conditions of this test, the test substance induced hematological effects with a modification of the structure of the spleen.
Executive summary:

Male and female rats (10 per sex per dose) were orally administered 5, 15 and 45 mg/kg bw/day of the test substance daily during 5 weeks. At the end of the exposure, half of the animals were sacrificed and the others were kept for 4 weeks without treatment to detect recovery from toxic effects. Animals were examined for hematological effects (haemoglobin concentraion, hematocrit, platelet count, erythrocyte count). The test substance induced mainly hematological effects with alteration of the spleen. Nevertheless, as soon as the treatment is finished all animals showed reversibility of the observed effects.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEL
5 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
The study is not GLP but a well conducted 5 weeks rat oral study.

Repeated dose toxicity: inhalation - systemic effects

Link to relevant study records
Reference
Endpoint:
short-term repeated dose toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 06 November To 08 March 1976
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study well conducted, sufficiently detailed, according to standardised guidelines, but not GLP.
Qualifier:
no guideline followed
Principles of method if other than guideline:
Male and female rats were exposed to the test substance for 4 hours per day, 6 days per week during 4 weeks. Following the exposure, a rest period of 4 weeks was performed. Hematology examinations were performed at 2, 4, 6 and 8 weeks. Clinical Biochemistry examinations were performed at the end of the exposure (4 weeks). Histology and necropsy were also performed at the end of the exposure.
GLP compliance:
no
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
No data
Route of administration:
inhalation: vapour
Type of inhalation exposure:
whole body
Vehicle:
air
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: Glass chamber
- Exposure chamber volume: 100 L with flow rate of 500L/h


OTHER:
Each day, the product was diluted in ethanol (CAS: 64-17-5)
- Solution (3) = 130 mg/m3 (0.050 ml/heure)
Product 4 ml + ethanol 11.2 ml
- Solution (2) = 40 mg/m3 (0.015 ml/heure)
Product 1.5 ml + ethanol 17.5 ml
- Solution (1) = 130 mg/m3 (0.005 ml/heure)
Dilution 1/10 of solution 3
solution (3) 1 ml + ethanol 9 ml
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
4 hours
Frequency of treatment:
6 days per week for 4 weeks
Remarks:
Doses / Concentrations:
13, 40 and 130 mg/m3 (0.013, 0.04 and 0.13 mg/l)
Basis:
nominal conc.
No. of animals per sex per dose:
8 per dose per sex
Control animals:
yes, concurrent vehicle
Details on study design:
Exposure time: 4 hours, 6 days a week and for 4 weeks, with 4 weeks of reversibility.
After 4 weeks of treatment, 5 males and 5 females per dose were sacrified in order to examine haematology (hemoglobinemia, numeration, erythrocytes and leucocytes), biochemistry (urea and bilirubin) and necropsy.
Other animals were observed during 4 weeks (reversibility).
Haematology was examined after 2 and 4 weeks for animals sacrified after 4 weeks, after 6 and 8 weeks for the others.
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: No

DETAILED CLINICAL OBSERVATIONS: No

BODY WEIGHT: No

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: at 2, 4, 6 and 8 weeks
- Anaesthetic used for blood collection: yes
- Animals fasted: No data
- How many animals: 5 per sex at the end of exposure/ 3 per sex after a week of rest.
- Parameters checked in table [No.1] were examined.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at the end of the exposure (4 weeks).
- Animals fasted: No data
- How many animals: 5 per sex.
- Parameters checked in table [No.1] were examined.

URINALYSIS: yes

NEUROBEHAVIOURAL EXAMINATION: No
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: YES (Histopatology of heart, lung, liver, spleen, kidneys and adrenal glands was performed)
Clinical signs:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
not examined
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:
not examined
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Details on results:
CLINICAL SIGNS AND MORTALITY
No mortality related to test substance was observed and no clinical signs were reported. One female died due to an anesthesic accident.

HAEMATOLOGY
After 2 weeks of treatment, no effect was observed in all animals. At the end of the exposure (4 weeks), a slight decrease in the erythrocytes count and a significant decrease of leucocytes count at the highest tested doses were observed. In addition, the methemoglobin increased at 40 mg/m3 (3.3 %) and became anormal at the highest tested dose (4.5 %). After two weeks of rest, all parameters showed comparable values to controls. (For details see Table 1)

CLINICAL CHEMISTRY
No significant effect on clinical biochemistry parameters (sodium, potassium, urea, total protein, alkaline phosphatase and alanine aminotransferase) was observed. (For details see Table 2)


GROSS PATHOLOGY
No effect has been observed.

HISTOPATHOLOGY
An effect of the test substance on the spleen was observed. The number of foci of extramedullary hematopoiesis were increased in the red pulpe of the spleen and were accompanied by an increase of splenic hemosiderin and venous sinus congestion. After 4 weeks of rest, foci of hematopoiesis have decreased in all animals. Only clusters of hemosiderin in macrophages were persistant.
Key result
Dose descriptor:
NOAEC
Effect level:
0.013 mg/L air (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Effects on spleen
Key result
Critical effects observed:
yes
Lowest effective dose / conc.:
0.013 mg/L air (nominal)
System:
haematopoietic
Organ:
blood
spleen
Treatment related:
yes
Dose response relationship:
yes
Relevant for humans:
yes

Table 1: Hematologic results

Time Groups H
%
Hb
g/100 ml
Erytrocyte
E+03/mm3
Leucocytes/mm3 Methemoglobin
After 2 weeks Control 45.2
2.17
15.0
0.23
7788
584.6
11880
3565.4
/
Control (ethanol) 44.3
2.34
-0.63
15.0
0.55
-0.08
7750
245.2
-0.15
9466
2498.5
-1.32
/
13 mg/m3 42.7
1.97
-2.03
14.7
0.76
-0.73
7113
491.5
-2.1
10933
4233.0
-0.39
/
40 mg/m3 46.0
1.54
0.71
15.5
0.52
2.1
7503
404.7
-0.95
12767
2834.0
0.46
/
130 mg/m3 44.1
2.40
-0.74
14.7
0.86
-0.65
7450
395.7
-1.1
12917
2324.0
0.53
/
After 4 weeks Control 49.3
2.9
15.8
1.0
8777
886
19289
33250
2.36
0.45
Control (ethanol) 48.3
1.88
-0.93
15.3
0.87
-1.13
8494
552
-0.85
15070
2865
-2.97
2.57
0.35
1.09
13 mg/m3 46.6 (-)
1.90
-2.44
15.0
0.76
-1.96
7940 (-)
642
-2.38
13930 (--)
2735
-3.85
2.87 (+)
0.53
2.35
40 mg/m3 46.9
2.38
-2.0
14.8 (-)
0;69
-2.4
8004 (-)
436
-2.46
12570 (---)
1865
-5.50
3.25
0.41
4.65 (+++)
130 mg/m3 45.2 (--)
2.1
-3.57
14.4 (-)
0.99
-2.97
7724 (-)
676
-2.93
11510 (---)
1995
-6.26
4.54
0.91
6.76 (+++)
After 6 weeks Control 45.3
4.3
15;6
0.23
8183
368
9983
1762
/
Control (ethanol) 50.0
1.8
1.77
16.0
0.34
2.08
7866
457
-1.32
10516
2125
0.47
/
13 mg/m3 49.0
1.8
1.94
15.9
0.64
0.83
7953
573
-0.83
11867
4694
0.92
/
40 mg/m3 49.8
2.9
2.14
15.3
0.56
-1.21
2270
423
0.38
10767
1261
0.89
/
130 mg/m3 49.3
2.7
1.93
15.5
0.27
-0.68
8133
242
-0.28
9500
1445
0.52
/
After 8 weeks Control / / 7753
380
/ 2.58
0.62
Control (ethanol) / / 7460
535
-1.06
/ /
13 mg/m3 / / 7810
460
0.23
/ /
40 mg/m3 / / 8013
373
1.19
/ /
130 mg/m3 / / 7670
259
-0.44
/ 1.97
0.30
-2.20

Statistically significant from control at : 95 % (+ or -), 99 % ( ++ or --), 99.9 % (+++ or ---)

Table 2: Chemical biochemistry results

Groups Na mEq/l K mEq/l Urea g/l Bilirubin mg/l Proteins g/l alkaline phosphatase alanine aminotransferase
Control 147.0
2.2
6.1
0.8
0.44
0.10
0.47
0.28
61.7
4.0
331
89
94*
62
Control (ethanol) 147.0
2.0
6.0
0.6
0.45
0.17
0.34
0.23
61.2
1.2
346
36
58
18
13 mg/m3 147.6
2.2
5.4
0.3
0.40
0.12
0.80
0.30
59.0
4.3
322
91
50
7
40 mg/m3 148.5
2.1
6.2
0.4
0.42
0.07
0.81
0.13
56.0
7.1
301
27
104*
151
130 mg/m3 149.0
4.1
5.6
0.4
0.46
0.07
0.97
1.34
56.6
4.2
296
31
70
44

* Elevated medium due to an outlier.

Conclusions:
Under the conditions of this test, the test substance showed a toxic effect at the blood cell level.
Executive summary:

Male and female rats were exposed to the test substance for 4 hours per day, 6 days per week during 4 weeks. Following the exposure, a rest period of 4 weeks was performed. Hematology examinations were performed at 2, 4, 6 and 8 weeks. Clinical Biochemistry examinations were performed at the end of the exposure (4 weeks). Histology and necropsy were also performed at the end of the exposure. Animals were exposed at 13, 40 or 130 mg/m3 of the test substance. No mortality related to test substance was observed and no clinical signs were reported. One female died due to an anesthesic accident. After 2 weeks of treatment, no effect was observed in all animals. At the end of the exposure (4 weeks), a slight decrease in the erythrocytes count and a significant decrease of leucocytes count at the highest tested doses were observed. In addition, the methemoglobin increased at 40 mg/m3 (3.3 %) and became anormal at the highest tested dose (4.5 %). After two weeks of rest, all parameters showed comparable values to controls. No significant effect on clinical biochemistry parameters (sodium, potassium, urea, total protein, alkaline phosphatase and alanine aminotransferase) was observed. No gross pathologic effect has been observed. An effect of the test substance on the spleen was observed. The number of foci of extramedullary hematopoiesis were increased in the red pulpe of the spleen and were accompanied by an increase of splenic hemosiderin and venous sinus congestion. After 4 weeks of rest, foci of hematopoiesis have decreased in all animals. Only clusters of hemosiderin in macrophages were persistant. Under the conditions of this test, the test substance showed a toxic effect at the blood cell level.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEC
13 mg/m³
Study duration:
subacute
Species:
rat
Quality of whole database:
The study is not GLP but a well conducted 4 weeks rat inhalation (vapor) study.

Repeated dose toxicity: inhalation - local effects

Link to relevant study records
Reference
Endpoint:
short-term repeated dose toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 06 November To 08 March 1976
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study well conducted, sufficiently detailed, according to standardised guidelines, but not GLP.
Qualifier:
no guideline followed
Principles of method if other than guideline:
Male and female rats were exposed to the test substance for 4 hours per day, 6 days per week during 4 weeks. Following the exposure, a rest period of 4 weeks was performed. Hematology examinations were performed at 2, 4, 6 and 8 weeks. Clinical Biochemistry examinations were performed at the end of the exposure (4 weeks). Histology and necropsy were also performed at the end of the exposure.
GLP compliance:
no
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
No data
Route of administration:
inhalation: vapour
Type of inhalation exposure:
whole body
Vehicle:
air
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: Glass chamber
- Exposure chamber volume: 100 L with flow rate of 500L/h


OTHER:
Each day, the product was diluted in ethanol (CAS: 64-17-5)
- Solution (3) = 130 mg/m3 (0.050 ml/heure)
Product 4 ml + ethanol 11.2 ml
- Solution (2) = 40 mg/m3 (0.015 ml/heure)
Product 1.5 ml + ethanol 17.5 ml
- Solution (1) = 130 mg/m3 (0.005 ml/heure)
Dilution 1/10 of solution 3
solution (3) 1 ml + ethanol 9 ml
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
4 hours
Frequency of treatment:
6 days per week for 4 weeks
Remarks:
Doses / Concentrations:
13, 40 and 130 mg/m3 (0.013, 0.04 and 0.13 mg/l)
Basis:
nominal conc.
No. of animals per sex per dose:
8 per dose per sex
Control animals:
yes, concurrent vehicle
Details on study design:
Exposure time: 4 hours, 6 days a week and for 4 weeks, with 4 weeks of reversibility.
After 4 weeks of treatment, 5 males and 5 females per dose were sacrified in order to examine haematology (hemoglobinemia, numeration, erythrocytes and leucocytes), biochemistry (urea and bilirubin) and necropsy.
Other animals were observed during 4 weeks (reversibility).
Haematology was examined after 2 and 4 weeks for animals sacrified after 4 weeks, after 6 and 8 weeks for the others.
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: No

DETAILED CLINICAL OBSERVATIONS: No

BODY WEIGHT: No

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: at 2, 4, 6 and 8 weeks
- Anaesthetic used for blood collection: yes
- Animals fasted: No data
- How many animals: 5 per sex at the end of exposure/ 3 per sex after a week of rest.
- Parameters checked in table [No.1] were examined.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at the end of the exposure (4 weeks).
- Animals fasted: No data
- How many animals: 5 per sex.
- Parameters checked in table [No.1] were examined.

URINALYSIS: yes

NEUROBEHAVIOURAL EXAMINATION: No
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: YES (Histopatology of heart, lung, liver, spleen, kidneys and adrenal glands was performed)
Clinical signs:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
not examined
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:
not examined
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Details on results:
CLINICAL SIGNS AND MORTALITY
No mortality related to test substance was observed and no clinical signs were reported. One female died due to an anesthesic accident.

HAEMATOLOGY
After 2 weeks of treatment, no effect was observed in all animals. At the end of the exposure (4 weeks), a slight decrease in the erythrocytes count and a significant decrease of leucocytes count at the highest tested doses were observed. In addition, the methemoglobin increased at 40 mg/m3 (3.3 %) and became anormal at the highest tested dose (4.5 %). After two weeks of rest, all parameters showed comparable values to controls. (For details see Table 1)

CLINICAL CHEMISTRY
No significant effect on clinical biochemistry parameters (sodium, potassium, urea, total protein, alkaline phosphatase and alanine aminotransferase) was observed. (For details see Table 2)


GROSS PATHOLOGY
No effect has been observed.

HISTOPATHOLOGY
An effect of the test substance on the spleen was observed. The number of foci of extramedullary hematopoiesis were increased in the red pulpe of the spleen and were accompanied by an increase of splenic hemosiderin and venous sinus congestion. After 4 weeks of rest, foci of hematopoiesis have decreased in all animals. Only clusters of hemosiderin in macrophages were persistant.
Key result
Dose descriptor:
NOAEC
Effect level:
0.013 mg/L air (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Effects on spleen
Key result
Critical effects observed:
yes
Lowest effective dose / conc.:
0.013 mg/L air (nominal)
System:
haematopoietic
Organ:
blood
spleen
Treatment related:
yes
Dose response relationship:
yes
Relevant for humans:
yes

Table 1: Hematologic results

Time Groups H
%
Hb
g/100 ml
Erytrocyte
E+03/mm3
Leucocytes/mm3 Methemoglobin
After 2 weeks Control 45.2
2.17
15.0
0.23
7788
584.6
11880
3565.4
/
Control (ethanol) 44.3
2.34
-0.63
15.0
0.55
-0.08
7750
245.2
-0.15
9466
2498.5
-1.32
/
13 mg/m3 42.7
1.97
-2.03
14.7
0.76
-0.73
7113
491.5
-2.1
10933
4233.0
-0.39
/
40 mg/m3 46.0
1.54
0.71
15.5
0.52
2.1
7503
404.7
-0.95
12767
2834.0
0.46
/
130 mg/m3 44.1
2.40
-0.74
14.7
0.86
-0.65
7450
395.7
-1.1
12917
2324.0
0.53
/
After 4 weeks Control 49.3
2.9
15.8
1.0
8777
886
19289
33250
2.36
0.45
Control (ethanol) 48.3
1.88
-0.93
15.3
0.87
-1.13
8494
552
-0.85
15070
2865
-2.97
2.57
0.35
1.09
13 mg/m3 46.6 (-)
1.90
-2.44
15.0
0.76
-1.96
7940 (-)
642
-2.38
13930 (--)
2735
-3.85
2.87 (+)
0.53
2.35
40 mg/m3 46.9
2.38
-2.0
14.8 (-)
0;69
-2.4
8004 (-)
436
-2.46
12570 (---)
1865
-5.50
3.25
0.41
4.65 (+++)
130 mg/m3 45.2 (--)
2.1
-3.57
14.4 (-)
0.99
-2.97
7724 (-)
676
-2.93
11510 (---)
1995
-6.26
4.54
0.91
6.76 (+++)
After 6 weeks Control 45.3
4.3
15;6
0.23
8183
368
9983
1762
/
Control (ethanol) 50.0
1.8
1.77
16.0
0.34
2.08
7866
457
-1.32
10516
2125
0.47
/
13 mg/m3 49.0
1.8
1.94
15.9
0.64
0.83
7953
573
-0.83
11867
4694
0.92
/
40 mg/m3 49.8
2.9
2.14
15.3
0.56
-1.21
2270
423
0.38
10767
1261
0.89
/
130 mg/m3 49.3
2.7
1.93
15.5
0.27
-0.68
8133
242
-0.28
9500
1445
0.52
/
After 8 weeks Control / / 7753
380
/ 2.58
0.62
Control (ethanol) / / 7460
535
-1.06
/ /
13 mg/m3 / / 7810
460
0.23
/ /
40 mg/m3 / / 8013
373
1.19
/ /
130 mg/m3 / / 7670
259
-0.44
/ 1.97
0.30
-2.20

Statistically significant from control at : 95 % (+ or -), 99 % ( ++ or --), 99.9 % (+++ or ---)

Table 2: Chemical biochemistry results

Groups Na mEq/l K mEq/l Urea g/l Bilirubin mg/l Proteins g/l alkaline phosphatase alanine aminotransferase
Control 147.0
2.2
6.1
0.8
0.44
0.10
0.47
0.28
61.7
4.0
331
89
94*
62
Control (ethanol) 147.0
2.0
6.0
0.6
0.45
0.17
0.34
0.23
61.2
1.2
346
36
58
18
13 mg/m3 147.6
2.2
5.4
0.3
0.40
0.12
0.80
0.30
59.0
4.3
322
91
50
7
40 mg/m3 148.5
2.1
6.2
0.4
0.42
0.07
0.81
0.13
56.0
7.1
301
27
104*
151
130 mg/m3 149.0
4.1
5.6
0.4
0.46
0.07
0.97
1.34
56.6
4.2
296
31
70
44

* Elevated medium due to an outlier.

Conclusions:
Under the conditions of this test, the test substance showed a toxic effect at the blood cell level.
Executive summary:

Male and female rats were exposed to the test substance for 4 hours per day, 6 days per week during 4 weeks. Following the exposure, a rest period of 4 weeks was performed. Hematology examinations were performed at 2, 4, 6 and 8 weeks. Clinical Biochemistry examinations were performed at the end of the exposure (4 weeks). Histology and necropsy were also performed at the end of the exposure. Animals were exposed at 13, 40 or 130 mg/m3 of the test substance. No mortality related to test substance was observed and no clinical signs were reported. One female died due to an anesthesic accident. After 2 weeks of treatment, no effect was observed in all animals. At the end of the exposure (4 weeks), a slight decrease in the erythrocytes count and a significant decrease of leucocytes count at the highest tested doses were observed. In addition, the methemoglobin increased at 40 mg/m3 (3.3 %) and became anormal at the highest tested dose (4.5 %). After two weeks of rest, all parameters showed comparable values to controls. No significant effect on clinical biochemistry parameters (sodium, potassium, urea, total protein, alkaline phosphatase and alanine aminotransferase) was observed. No gross pathologic effect has been observed. An effect of the test substance on the spleen was observed. The number of foci of extramedullary hematopoiesis were increased in the red pulpe of the spleen and were accompanied by an increase of splenic hemosiderin and venous sinus congestion. After 4 weeks of rest, foci of hematopoiesis have decreased in all animals. Only clusters of hemosiderin in macrophages were persistant. Under the conditions of this test, the test substance showed a toxic effect at the blood cell level.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Study duration:
subacute
Species:
rat
Quality of whole database:
The study is not GLP but a well conducted 4 weeks rat inhalation (vapor) study.

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Justification for classification or non-classification

Based on the classification criteria of EU Regulation 1272/2008 (CLP), with an oral NOAEL=5 mg/kg bw/day and an inhalation NOAEC of 13 mg/m3, 1-fluoro-2nitrobenzene is classified as Single Target Organ Toxicant – Repeated Exposure category 1 (STOT-RE cat.1) with blood and spleen as target organs.

Data are only available for oral and inhalation studies, however, dermal route cannot be excluded and no specific route can be specified.