Registration Dossier

Administrative data

Endpoint:
sub-chronic toxicity: inhalation
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
1986
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study conducted to OECD guidelines and to GLP, and therefore meets the criteria for Klimisch code 1. Study is classified as Klimisch code 2 only because it is based on read across.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1986
Report Date:
1987

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 412 (Subacute Inhalation Toxicity: 28-Day Study)
Deviations:
no
GLP compliance:
yes
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
The test item belongs to the same chemical group as the submission substance (aryl/alkyl sulfonates)

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River
- Age at study initiation: males, 6 weeks; females, 7 weeks
- Weight at study initiation: males, 205-232g; females 156-186g
- Housing: suspended wire mesh cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 2 weeks


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21-26°C
- Humidity (%): 20-76%
- Photoperiod (hrs dark / hrs light): 12 hours light/dark cycle

Administration / exposure

Route of administration:
inhalation: aerosol
Type of inhalation exposure:
whole body
Vehicle:
other: unchanged (no vehicle)
Remarks on MMAD:
MMAD / GSD: MMAD 3.3-3.7 µm
GSD 2.0-2.1 µm
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus:Glass and stainless steel exposure chambers
- System of generating particulates/aerosols:
- Temperature, humidity, pressure in air chamber: 21-26°C, 20-76%
- Air flow rate: 210-215 pm
- Air change rate: 4.8/minute
- Method of particle size determination: Delron DCI-6 cascade impactor


TEST ATMOSPHERE
- Samples taken from breathing zone: yes

Analytical verification of doses or concentrations:
no
Details on analytical verification of doses or concentrations:
Concentration measured by gravimetric analysis.
Duration of treatment / exposure:
6 hours per day for 28 days.
Frequency of treatment:
5 days per week.
Doses / concentrations
Remarks:
Doses / Concentrations:
49.5, 156, 260 mg/m3
Basis:
analytical conc.
No. of animals per sex per dose:
5
Control animals:
yes, concurrent no treatment
Positive control:
None.

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily


DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: twice daily


BODY WEIGHT: Yes
- Time schedule for examinations: 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: Prior to terminal sacrifice
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: All animals
- Parameters examined: Haemoglobin concentration, haematocrit, erythrocyte count, platelet count, clotting time, total and differential leukocyte count


CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Prior to terminal sacrifice
- Animals fasted: No data
- How many animals: All animals



URINALYSIS: Yes
- Time schedule for collection of urine: Prior to terminal sacrifice
- Metabolism cages used for collection of urine: No data
- Animals fasted: No data



NEUROBEHAVIOURAL EXAMINATION: No

Sacrifice and pathology:
gross pathology on all animals
GROSS PATHOLOGY: Yes (see table)
HISTOPATHOLOGY: Yes (see table)
Statistics:
ANOVA with Dunnets test
Regression analysis
Krusal-Walis and Dunns summed rank test
Jonsheere's test for monotonic trend

Results and discussion

Results of examinations

Clinical signs:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Histopathological findings: neoplastic:
no effects observed
Details on results:

CLINICAL SIGNS AND MORTALITY
No mortality occurred during the test.

BODY WEIGHT AND WEIGHT GAIN
Bodyweight gain was decreased in treated animals, although this was not statistically significant.

FOOD CONSUMPTION
Not measured

FOOD EFFICIENCY
Not measured

WATER CONSUMPTION
Not measured

OPHTHALMOSCOPIC EXAMINATION
Not measured

HAEMATOLOGY
Statistically significant differences were observed in females. Specifically; increased haematocrit in the low dose group.
CLINICAL CHEMISTRY
Statistically significant differences were observed in females. Specifically; increased creatine phospholinase in the mid and top dose groups and sodium in the top dose group.

NEUROBEHAVIOUR
Not measured

ORGAN WEIGHTS
Statistically significant increased lung weights and lung to body weight ratios were observed in a dose dependant manner in the mid and top dose groups.

GROSS PATHOLOGY
Gross lesions were sporadic and were not considered to be caused by the test article.

HISTOPATHOLOGY: NON-NEOPLASTIC
A higher incidence of accumulations of intraalveolar macrophages and hyperplasia/hypertrophy of bronchiole epithelium was seen in the lungs of the treated animals. These were dose related at the mid and top dose levels.

Effect levels

Dose descriptor:
NOAEL
Effect level:
49.5 mg/m³ air (analytical)
Sex:
male/female
Basis for effect level:
other: Statistically significant dose related increase in lung weight and relative lung weights with corresponding accumulations of intraalveolar macrophages and hyperplasia/hypertrophy of the bronchiole epithelium.

Target system / organ toxicity

Critical effects observed:
not specified

Any other information on results incl. tables

 

Table 1: Average body weights and body weight gains during 28 days of treatment

 

Analytical concentration (mg/L)

Body Weights (g)

Total Weight Gain

Week 0

Week 1

Week 2

Week 3

Week 4

g

% of control

Male

    0

 217

 249

 295

336

 369

 152

 

LCT

 216

248

294

331

359 

 143

 94

MCT

 215

247 

302 

346

384 

 169

 111

HCT

 215

243 

285 

319

347 

 132

 87

Female

    0

 171

 185

 206

223

 239

 68

 

LCT

 169

 185

 205

220

 232

 63

 93

MCT

 168

 181

 203

219

 229

 61

 90

HCT

 172

 186

 205

223

 238

 66

 97

 

Table 2 Selected haematology, clinical chemistry and pathology findings

 

Doses (unit)

0

50

150

250

0

50

150

250

male

female

Number of animals/group

5

5

5

5

5

5

5

5

Haematology(day x)

 

 

 

 

 

 

 

 

- RBC (TERA/L)

6.62

6.89 

6.66 

6.52 

6.60 

6.90 

6.59 

6.70

- MCV (FL)

 -

- HCT (L/L)

 46

47 

46 

43 

 46

48 

47 

44 

- HGB (MMOL/L)

 16.1

16.7 

16.1 

15.5 

15.5 

16.6 

16.1 

15.5 

- WBC (GIGA/L)

 13.9

13.7 

12.9 

10.5 

9.0 

12.8 

11.5 

13.8 

Blood chemistry(day x)

 

 

 

 

 

 

 

 

- sodium (MMOL/L)

 146

146 

147 

146 

 145

143 

144 

141 

- potassium (MMOL/L)

 4.4

4.4 

4.5 

4.3 

 4.3

4.3 

4.3 

4.3 

- chloride (MMOL/L)

 101

102 

102 

102 

 102

102 

102 

101 

- gobulin (G/L)

 2.2

2.3 

2.4 

2.0 

 2.1

2.1 

2.1 

2.0 

- cholesterol (MMOL/L)

 56

53 

 63

55 

 59

75 

60 

69 

- triglyceride (MMOL/L)

 45

39 

54 

35 

 30

24 

25 

25 

Pathology

 

 

 

 

 

 

 

 

Accumulation of intraalveolar macrophages

 3

5

5

Bronchiolar epithelium: hyperplasia/hypertrophy

 3

Table 3: Absolute and relative organ weights

 

 

Males

Females

DAILY DOSE
(units e.g. mg/kg bw/day)

0

50

150

250

0

50

150

250

NUMBER OF ANIMALS

 5

BODY WEIGHT (g)a

 335

323 

346 

313 

 213

204 

204 

208 

BRAIN

 

 

 

 

 

 

 

 

Absolute Weighta

g

 1.976

1.939 

2.032 

1.943 

 1.900

1.876 

1.856 

1.830 

Per Body Weighta

%

 5.93

6.03 

5.90 

6.21 

 8.96

9.20 

9.12 

8.84 

ADRENALS

 

 

 

 

 

 

 

 

Absolute Weighta

g

 0.051

0.052 

0.054 

0.054 

 0.068

0.069 

0.070 

0.060 

Per Body Weighta

%

 1.52

1.63 

1.56 

1.71 

 3.21

3.37 

3.44 

2.90 

Per Brain Weighta

%

 

 

 

 

 

 

 

 

HEART

 

 

 

 

 

 

 

 

Absolute Weighta

g

 1.220

 1.173

 1.179

 1.122

 0.803

0.801 

0.753 

0.793 

Per Body Weighta

%

 3.64

3.65 

3.41 

3.59 

 3.79

3.92 

3.70 

3.82 

Per Brain Weighta

%

 

 

 

 

 

 

 

 

KIDNEYS

 

 

 

 

 

 

 

 

Absolute Weighta

g

 2.592

2.779 

3.054 

2.704 

 1.745

1.696 

1.694 

1.883 

Per Body Weighta

%

 7.79

8.61 

8.81 

8.61 

 8.16

8.32 

8.29 

9.05 

Per Brain Weighta

%

 

 

 

 

 

 

 

 

LIVER

 

 

 

 

 

 

 

 

Absolute Weighta

g

 10.775

10.306 

11.470 

9.999 

 7.020

6.464 

6.789 

7.357 

Per Body Weighta

%

 3.22

3.20 

3.31 

3.18 

 3.30

3.17 

3.33 

3.53 

Per Brain Weighta

%

 

 

 

 

 

 

 

 

SPLEEN

 

 

 

 

 

 

 

 

Absolute Weighta

g

 0.670

0.569 

0.652 

0.594 

 0.426

0.441 

0.471 

0.484 

Per Body Weighta

%

 2.00

 1.77

1.88 

1.89 

 2.01

2.17 

2.30 

2.34 

Per Brain Weighta

%

 

 

 

 

 

 

 

 

TESTES

 

 

 

 

n.a.b

n.a.b

n.a.b

n.a.b

Absolute Weighta

g

 3.187

3.072 

2.796 

3.135 

n.a.b

n.a.b

n.a.b

n.a.b

Per Body Weighta

%

 9.54

 9.54

8.16 

10.02 

n.a.b

n.a.b

n.a.b

n.a.b

Per Brain Weighta

%

 

 

 

 

n.a.b

n.a.b

n.a.b

n.a.b

LUNGS

 

 

 

 

 

 

 

 

Absolute Weighta

g

 1.306

1.302 

1.515 

1.537 

 1.051

1.127 

1.138 

1.338 

Per Body Weighta

%

 3.91

 4.05

4.39 

4.91 

 4.93

5.52 

5.59 

6.46 

Per Brain Weighta

%

 

 

 

 

 

 

 

 

OVARIES

n.a.b

n.a.b

n.a.b

n.a.b

 0.0921

0.0768 

0.0943 

0.0742 

Absolute Weighta

g

n.a.b

n.a.b

n.a.b

n.a.b

 4.34

3.77 

4.62 

3.56 

Per Body Weighta

%

n.a.b

n.a.b

n.a.b

n.a.b

 

 

 

 

Per Brain Weighta

%

n.a.b

n.a.b

n.a.b

n.a.b

 

 

 

 

 

Applicant's summary and conclusion

Conclusions:
A NOAEL of 49.5 mg/m³ was identified for males and females, based on increased lung weight and microscopic changes of the lung.
Executive summary:

In a subacute inhalation toxicity study, a petroleum derived calcium salt was administered to 5 Sprague-Dawley rats/sex/concentration by whole body exposure at concentrations of 0, 49.5, 156 or 260 mg/m³ for 6 hours per day, 5 days/week for a total of 28 days.

 

Statistically significant, dose related increases in lung weights occurred in the mid and top dose groups, with corresponding increases in intraalveolar macrophages and hyperplasia/hypertrophy of bronchiole epithelium.  The LOAEL is 156 mg/m³, based on effects in the lung. The NOAEL is 49.5 mg/m³.