Benzene mono-C10-13-alkyl derivatives, distillation residues, sulfonated, sodium salts

Repeated dose toxicity: inhalation

Administrative data

Endpoint:

sub-chronic toxicity: inhalation

Type of information:

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

Adequacy of study:

supporting study

Study period:

1987

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results. The study report was conclusive, done to a valid guideline and the study was conducted under GLP conditions. In addition, as this study is used in the context of a read across, Klimisch 2 is assigned.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1987

Report date:

1987

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 412 (Subacute Inhalation Toxicity: 28-Day Study)

Deviations:

no

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

61789-86-4

IUPAC Name:

61789-86-4

Constituent 2

Reference substance name:

Sulfonic acids, petroleum, calcium salts

IUPAC Name:

Sulfonic acids, petroleum, calcium salts

Constituent 3

Reference substance name:

Sulfonic acids, petroleum, calcium salts

EC Number:

263-093-9

EC Name:

Sulfonic acids, petroleum, calcium salts

Cas Number:

61789-86-4

IUPAC Name:

263-093-9

Test material form:

other: liquid

Details on test material:

Details on test material
Sponsor's identification: OS80020 (product as manufactured diluted to 65% in mineral oil)
Description: Brown liquid
Purity: 35% Active Ingredient (contains 65% oil)
Date received 03 October 1986
Storage conditions: Temperature monitored room (60 - 85°F).
Expiry date: Not supplied

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system 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:

air

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, sham-exposed

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	5	5	5	5	5
Bronchiolar epithelium: hyperplasia/hypertrophy	3	4	5	5	4	5	5	5

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	5	5	5	5	5	5	5
BODY WEIGHT (g)^a		335	323	346	313	213	204	204	208
BRAIN
Absolute Weight^a	g	1.976	1.939	2.032	1.943	1.900	1.876	1.856	1.830
Per Body Weight^a	%	5.93	6.03	5.90	6.21	8.96	9.20	9.12	8.84
ADRENALS
Absolute Weight^a	g	0.051	0.052	0.054	0.054	0.068	0.069	0.070	0.060
Per Body Weight^a	%	1.52	1.63	1.56	1.71	3.21	3.37	3.44	2.90
Per Brain Weight^a	%
HEART
Absolute Weight^a	g	1.220	1.173	1.179	1.122	0.803	0.801	0.753	0.793
Per Body Weight^a	%	3.64	3.65	3.41	3.59	3.79	3.92	3.70	3.82
Per Brain Weight^a	%
KIDNEYS
Absolute Weight^a	g	2.592	2.779	3.054	2.704	1.745	1.696	1.694	1.883
Per Body Weight^a	%	7.79	8.61	8.81	8.61	8.16	8.32	8.29	9.05
Per Brain Weight^a	%
LIVER
Absolute Weight^a	g	10.775	10.306	11.470	9.999	7.020	6.464	6.789	7.357
Per Body Weight^a	%	3.22	3.20	3.31	3.18	3.30	3.17	3.33	3.53
Per Brain Weight^a	%
SPLEEN
Absolute Weight^a	g	0.670	0.569	0.652	0.594	0.426	0.441	0.471	0.484
Per Body Weight^a	%	2.00	1.77	1.88	1.89	2.01	2.17	2.30	2.34
Per Brain Weight^a	%
TESTES						n.a.^b	n.a.^b	n.a.^b	n.a.^b
Absolute Weight^a	g	3.187	3.072	2.796	3.135	n.a.^b	n.a.^b	n.a.^b	n.a.^b
Per Body Weight^a	%	9.54	9.54	8.16	10.02	n.a.^b	n.a.^b	n.a.^b	n.a.^b
Per Brain Weight^a	%					n.a.^b	n.a.^b	n.a.^b	n.a.^b
LUNGS
Absolute Weight^a	g	1.306	1.302	1.515	1.537	1.051	1.127	1.138	1.338
Per Body Weight^a	%	3.91	4.05	4.39	4.91	4.93	5.52	5.59	6.46
Per Brain Weight^a	%
OVARIES		n.a.^b	n.a.^b	n.a.^b	n.a.^b	0.0921	0.0768	0.0943	0.0742
Absolute Weight^a	g	n.a.^b	n.a.^b	n.a.^b	n.a.^b	4.34	3.77	4.62	3.56
Per Body Weight^a	%	n.a.^b	n.a.^b	n.a.^b	n.a.^b
Per Brain Weight^a	%	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.

 

This subchronic toxicity study in the rat is acceptable and satisfies the guideline requirement (OECD 412) for a subchronic inhalation study in the rat.