Benzenesulfonic acid, mono- and dialkylation products with C16-20 (even numbered, branched and linear) olefins, calcium salts, calcium carbonate, overbased, including distillates (petroleum), hydrotreated, solvent-refined, solvent-dewaxed, or catalytic dewaxed, light or heavy paraffinic C15-C50

Endpoint summary

Description of key information

The toxicity of the substance has been assessed  by the repeated exposure over a period of 28-days by the three routes of exposure, oral, dermal and inhalation using read across studies .

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:

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

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study conducted to OECD guidelines to to GLP, and therefore meets the requirements for Klimisch code 1. Study is classified as Klimisch code 2 only because it is based on read across.

Qualifier:

according to guideline

Guideline:

OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)

Deviations:

yes

Remarks:

A fuctional observation battery for neurotoxicity was not performed since this test was not par of the OECD 407 guideline at the time the study was performed

GLP compliance:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories
- Age at study initiation: 41 days

Route of administration:

oral: gavage

Vehicle:

peanut oil

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS: Mixed weekly weight/volume in peanut oil.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Chemical analysis of dosing solutions was conducted to confirm that they were homogeneous and met the desired concentrations.

Duration of treatment / exposure:

29 day treatment duration with a 14 day recovery period.

Frequency of treatment:

7 days/week

Remarks:

Doses / Concentrations:
gavage doses of 0, 100, 500 or 1000 mg/kg/bw
Basis:
actual ingested

No. of animals per sex per dose:

6

Control animals:

yes

Details on study design:

- dose selection rationale: Data from a pilot two week repeated dose oral study.

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Twice daily

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes

Statistics:

Tests applied included; parametric ANOVA with Dunnetts post-hoc test, non parametric Kruskal-Walls and Mann-Whitney U test, Bartletts test for equal variances, Students t test and Dixons test for rejection of outlying values.

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 specified

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

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:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Details on results:

CLINICAL SIGNS AND MORTALITY
One animal was sacrificed on Day 0 and one animal was found dead on Day 9, result of probable misdosing.

Dose descriptor:

NOAEL

Effect level:

500 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Mean serum cholestrol levels were significantly reduced in the 1000 mg/kg males and females at termination of dosing. This was still significantly reduced in 1000 mg/kg females at the end of the 14 day recovery period.

Critical effects observed:

not specified

 

Table 1: Average body weights and body weight gains during 29 days of treatment

 

Dose rate (mg/kg	Body Weights (g)					Total Weight Gain
	Week 0	Week 1	Week 2	Week 3	Week 4	g	% of control
Male
0	195	243	286	323	352	157	N/A
100	196	245	298	340	374	178	111
500	200	245	289	329	362	162	103
1000	193	240	283	315	346	153	97
Female
0	155	175	194	213	223	68	N/A
100	156	174	194	215	228	72	106
500	154	175	190	212	221	67	99
1000	155	174	195	212	224	69	101

 

Conclusions:

A NOAEL of 500 mg/kg bw/day was identified in this study.

Executive summary:

In a subchronic toxicity study calcium sulphonate was administered to 12 Sprague-Dawley rats/sex/dose in the control and top dose groups and 6 animals Sprague-Dawley rats/sex/dose in the low and mod dose via gavage at dose levels of 0, 100, 500 or 1000 mg/kg bw/day). A decrease in serum cholesterol levels occurred in the top dose group. The LOAEL is 1000 mg/kg bw/day, based on a decrease in serum cholesterol at the top dose. The NOAEL is 500 mg/kg bw/day. This subchronic toxicity study in the rat is acceptable and satisfies the guideline requirement for a subchronic oral study (OPPTS 870.3100; OECD 408) in rats.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

NOAEL

500 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

There are two Klimisch score 1 read across studies avaiable.

Repeated dose toxicity: inhalation - systemic effects

Link to relevant study records

Reference

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.

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes

Limit test:

no

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

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.

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.

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

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.

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.

Critical effects observed:

not specified

 

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

 

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.

 

 

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

NOAEC

49.5 mg/m³

Study duration:

subacute

Species:

rat

Quality of whole database:

The read across study has a Klimisch score of 1.

Repeated dose toxicity: dermal - systemic effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: dermal

Type of information:

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

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study meets the criteria for Klimisch code 1. Study is classified as Klimisch code 2 only because it is based on read across.

Qualifier:

according to guideline

Guideline:

OECD Guideline 410 (Repeated Dose Dermal Toxicity: 21/28-Day Study)

Deviations:

no

GLP compliance:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

Sprague Dawley CD rats, 8-9 weeks of age at initiation

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on exposure:

test substance was applied undiluted to the clipped dorsal surface for periods of 6h per day of study. material was held in place by a gauze patch secured with tape. after each exposure period the treated area was wiped. the procedure was repeated daily for 28d.

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

6h per day for 28d

Frequency of treatment:

daily

Remarks:

Doses / Concentrations:
0, 100,300 and 1000 mg/kg bwt
Basis:
nominal per unit body weight

No. of animals per sex per dose:

5

Control animals:

yes, sham-exposed

Positive control:

none

Observations and examinations performed and frequency:

clinical observations daily
dermal reposes on days 0,1,4,7,11,14,18,21,25 and prior to blood collection on d28
body weight and food consumption during treatment and recovery
Hematology and clinical chemistry at termination of treatment and recovery
Microscopic examination on all animals

Sacrifice and pathology:

full range of evaluations performed

Statistics:

ANOVA with Dunnets test, Kruskal-Walis, Dunns summed rank test , Jonkheere test for monotonic trend, Students t test

Clinical signs:

no effects observed

Dermal irritation:

effects observed, treatment-related

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 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:

no effects observed

Behaviour (functional findings):

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Details on results:

No mortality occurred during the study. Low incidences of very slight erythema, desquamation and / or pinpoint scabbing were observed sporadically in all treated animals. All animals were free of edema during the study. Body weights and food consumption were unremakable. There were no treatment related differences in heamatology. Differences from control were noted for several heamatology parameters including a statistically significant increase in mean % of neutrophils of 300 and 1000 mg/kg females and a decrease in mean % of lymphocytes in the 1000mg/kg females compared to controls on d28. In the absence of differences from control in absolute white blood cell counts, these findings were not considered related to treatment. There was a statistically significant decrease in mean corpuscular haemoglobin concentration in the male satellite animals from d28-42. In the absence of other significant findings these small differences were not considered clinically significant. Serum chemistry values were unremakable. Gross mostmortem findings were considered incidental and unrelated to treatment. There were no alterations in organ weights that were attributed to treatment.There were no test material related microscopic findings noted in any group. Effects on the skin were seen in all groups including control but tended to increase in male treated animals and females in the 300 and 1000 mg.kg groups, indicating a mild irritant effect of the test article.

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Basis for effect level:

other: no adverse effects at any concentration tested

Critical effects observed:

not specified

Conclusions:

No evidence of systemic toxicity via the dermal route.

Executive summary:

the test article exhibited no evidence of systemic toxicity via the dermal route under the conditions of this study. A NOAEL of >1000 mg/kg bw was established.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

The NOEAL was selected from a Klimisch score 1 study. A lower quality study is also available.

Additional information

The repeat dose toxicity has been determined by subacute 28-day toxicity studies by oral, dermal and inhalation exposure. Based on the data available, the substance may have the potential for haemotological effects with a reduction in cholesterol observed in one of the studies by oral exposure. Affects observed in the repeat dose inhalation toxicity study available demonstrated enlarged lungs in high and intermediate dose animals which are likely to be physical effects due to the inhalation of mineral oil and not necessarily a direct toxicological effect of the registered substance.

Furthermore, one dermal repeat dose study exists noting effects which resulted in the Study Director being unable to assign a NOAEL. Due consideration of these data has been taken, but the results are considered not representative of the toxicological effects of the substance since the observations have not been repeated in the remaining study with dermal exposure, nor following exposure by oral and inhalation routes which are generally considered to represent greater systemic exposure routes. Furthermore, effects to the testes of male rats were not been observed in the reproductive toxicity study. In consequence, although data are not available to adequately determine the cause of these effects, the fact that no similar observations have been found in any other study undertaken is considered adequate justification to regard these data as not representative of the toxicological profile of the registered substance.

Notwithstanding these effects, no further effects were observed in the studies and each study achieved a NOAEL. The data are therefore considered adequate to assess the toxicological profile of the substance and for the purposes of classification.

Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
The NOAEL was selected from the most conservative value of the two available read-across studies.

Justification for selection of repeated dose toxicity inhalation - systemic effects endpoint:
There is only one read across study available.

Justification for selection of repeated dose toxicity inhalation - local effects endpoint:
Any local effects will be covered in the NOAEL for systemic effects.

Justification for selection of repeated dose toxicity dermal - systemic effects endpoint:
The NOAEL was selected from the higher quality subacute dermal study.

Justification for selection of repeated dose toxicity dermal - local effects endpoint:
All local effects will be covered in the NOAEL for systemic effects.

Repeated dose toxicity: via oral route - systemic effects (target organ) cardiovascular / hematological: other

Repeated dose toxicity: inhalation - systemic effects (target organ) respiratory: lung

Justification for classification or non-classification

In accordance with the criteria for classification as defined in Annex I, Regulation 1272/2008, the test material does not require classification for repeated dose toxicity.