Benzenesulfonic acid, 4-C10-13-sec-alkyl derivs., compds. with ethanolamine

Repeated dose toxicity: oral

Administrative data

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1972

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference Type:

publication

Title:

Unnamed

Year:

1972

Materials and methods

Principles of method if other than guideline:

The purpose of this study is to determine the sub chronic toxicity of C10 - C14 linear alkylbenzene sulfonic acid sodium salt (LAS) in Wistar SLC rats. Male and female rats were maintained on diets of 0, 0.07, 0.2, 0.6 and 1.8% LAS (equivalent to 0, 40, 115, 340, 1030 mg/kg bw/day) for 26 weeks. Clinical observations, water consumption, food consumption and body weights were recorded weekly. At the end of study, blood was collected for estimation of hematological and clinical chemistry parameters. Urinalysis was also performed 2 weeks before necropsy. All the surviving animals were humanely euthanized and gross necropsy, organs weights and histopathology were performed.

GLP compliance:

no

Remarks:

(pre-dates GLP)

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Benzenesulfonic acid, C10-14-alkyl derivs., sodium salts

EC Number:

274-070-8

EC Name:

Benzenesulfonic acid, C10-14-alkyl derivs., sodium salts

Cas Number:

69669-44-9

IUPAC Name:

sodium 4-dodecylbenzenesulfonate

Test material form:

solid

Remarks:

(powder)

Details on test material:

- Name of test material: Linear alkylbenzene sulfonic acid sodium salt (LAS); NeoPlex F-60
- Average molecular weight: 345.8 g
- Composition:: 60 ± 2% (active ingredient) of C10 - C14 LAS, <1.5% unreacted lipids, <4% inorganic salts and 38% moisture
- Lot no.: 127 (Supplier: Kao chemicals)
- Distribution of carbon count of the alkyl groups and component of each sample:
LAS: C10 (10.6%), C11 (34.1%), C12 (27.7%), C13 (19.0%) and C14 (8.7%)

No further details on test substance are provided in the study report.

Test animals

Species:

rat

Strain:

other: Wistar SLC

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Male and female were obtained from Shizuoka Agricultural Cooperative Association for Laboratory Animals
- Age at study initiation: Approximately 4 weeks
- Weight at study initiation: 80 - 90 g (male rats), 65 - 80 g (female rats)
- Fasting period before study: No
- Housing: 5 animals were housed per stainless steel cage.
- Diet: FII food (from Funabashi Farm); ad libitum
- Water: Tap water; ad libitum
- Acclimation period: Not reported

ENVIRONMENTAL CONDITIONS
- Temperature: 25 ± 1°C
- Humidity: 55 - 65%
- Air changes: Not reported
- Photoperiod: 12 hours dark /12 hours light

IN-LIFE DATES: From: May 30, 1972 To: Nov. 27, 1972

Administration / exposure

Route of administration:

oral: feed

Details on route of administration:

C10 - C14 LAS was administered to animals by mixing 0.07, 0.2, 0.6 and 1.8% with pulverized FII feed, the powders were shaped into solid form before being freely provided for feeding along with tap water. As control, solid FII feed was provided to the animals.

Vehicle:

other: C10 - C14 LAS was administered in diet

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

26 weeks

Frequency of treatment:

Continuous in diet (ad libitum)

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

0.07 other: %

Remarks:

in diet

Doses / concentrations 2

Dose / conc.:

0.2 other: %

Remarks:

in diet

Doses / concentrations 3

Dose / conc.:

0.6 other: %

Remarks:

in diet

Doses / concentrations 4

Dose / conc.:

1.8 other: %

Remarks:

in diet

Doses / concentrations 5

Dose / conc.:

40 mg/kg bw/day (nominal)

Remarks:

in diet (corresponding to 0.07% dose level)

Doses / concentrations 6

Dose / conc.:

115 mg/kg bw/day (nominal)

Remarks:

in diet (corresponding to 0.2% dose level)

Doses / concentrations 7

Dose / conc.:

340 mg/kg bw/day (nominal)

Remarks:

in diet (corresponding to 0.6% dose level)

Doses / concentrations 8

Dose / conc.:

1 030 mg/kg bw/day (nominal)

Remarks:

in diet (corresponding to 1.8% dose level)

No. of animals per sex per dose:

10 animals/sex/dose group

Control animals:

yes, plain diet

Positive control:

No

Examinations

Observations and examinations performed and frequency:

CLINICAL OBSERVATIONS: Yes

BODY WEIGHT: Yes
- Time schedule for examinations: Weekly

FOOD CONSUMPTION AND COMPOUND INTAKE:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Time schedule for examinations: Weekly

WATER CONSUMPTION AND COMPOUND INTAKE: Yes
- Time schedule for examinations: Weekly

OPHTHALMOSCOPIC EXAMINATION: No

HEMATOLOGY: Yes
- Time schedule for collection of blood: After 26 weeks, blood was collected from each animal prior to autopsy (for hemoglobin, hematocrit, red blood cells, white blood fractions and platelets) and during euthanasia (erythrocyte membrane resistance)
- Anesthetic used for blood collection: No
- Animals fasted: No
- How many animals: All surviving animals
- Parameters: Hemoglobin, hematocrit, red blood cells, white blood fractions and platelets, erythrocyte membrane resistance and blood coagulation time

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: After 26 weeks, blood was collected from each animal after euthanasia.
- Animals fasted: No
- How many animals: All surviving animals
- Parameters: Total proteins, GOT activity, ALP activity, LAP activity, urea nitrogen, A/G ratio and bilirubin

URINALYSIS: Yes
- Time schedule: 2 weeks prior to autopsy, urine isolated and collected from waste was analyzed.
- Metabolism cages used for collection of urine: No
- Animals fasted: No
- How many animals: All surviving animals
- Parameters: Proteins, glucose, ketone bodies, occult blood, bilirubin and urobilinogen

NEUROBEHAVIOURAL EXAMINATION: No

IMMUNOLOGY: No

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
Gross findings were obtained after euthanizing animals and macroscopic findings were observed.

ORGAN WEIGHTS: Brain, pituitary, thyroid, thymus, heart, lungs, liver, kidneys, spleen, adrenal gland, prostate, testes, uterus, ovaries and appendix

HISTOPATHOLOGY: Yes
Collected tissues along with stomach, large and small intestines, pancreas, epididymis, skin and bones) were weighed, examined and subjected to microscopy through H.E., PAS, lipid and silver staining.

Results and discussion

Results of examinations

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

Severe diarrhea was observed in the high concentration group (1.8%).

Mortality:

mortality observed, treatment-related

Description (incidence):

In 24th week, 1 male animal from the 1.8% dose group died.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

There was significant inhibition of weight gain in both males and females of 1.8% dose group.

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

Food consumption was low in the 1.8% dose group relative to the control.

Food efficiency:

not examined

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

effects observed, treatment-related

Description (incidence and severity):

Water consumption was slightly low in animals of 1.8% dose group relative to the control

Ophthalmological findings:

not examined

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

In females of 1.8% dose group, a significant decrease in hematocrit and hemoglobin and significant increase in the erythrocyte membrane resistance was observed.. For both males and female species, there was increase in white blood cell count at 1.8% dose.

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

There was dose dependent significant increase in ALP levels (at mid and high dose) and significant decrease in total protein (at high dose).

Urinalysis findings:

no effects observed

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

1.8% dose group: Significant increase in relative weights of liver, suprarenal gland, testis, brain and caecum were observed in males and relative weights of heart, thyroid, liver, uterus, brain and caecum of females.
0.6% dose group: Significant increase in caecum relative weights in males and hypophysis in females.
0.2% dose group: Caecum relative weights were significantly increased in males and kidney relative weights were significantly decreased in females.

Gross pathological findings:

effects observed, non-treatment-related

Description (incidence and severity):

Color of the liver in 2 males and 5 females from the 1.8% group was lighter than normal and seemed to have a yellowish white color mixed to it. In addition, lung tumors were also seen in one female rat from the 0.2% group.

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

The main findings were concerned with kidneys, liver and intestinal tract.
Kidney damage was seen at and above 0.2% dose groups. Mild hepatocyte changes were observed in animals of 1.8% dose group and microscopic changes in intestinal tract were observed in female rats of the 1.8 and 0.6% dose groups.

Histopathological findings: neoplastic:

not examined

Other effects:

not examined

Details on results:

CLINICAL SIGNS:
- Severe diarrhea was observed in animals in the high concentration groups (1.8%) immediately after starting the study, and although the diarrhea gradually became milder after about 1 week, the animal s did not recover fully by the end of the study. The animals moved slowly, and their lower abdomen was dirty.

MORTALITY: During week 24, 1 male animal from 1.8% dose group died.
BODY WEIGHT AND WEIGHT CHANGES: There was significant inhibition of weight gain in both males and females of 1.8% dose group, while the 0.6% dose groups showed slight but not significant trends to suppress weight gain.

FOOD CONSUMPTION AND COMPOUND INTAKE: Food consumption was low in the 1.8% dose group relative to the control.

WATER CONSUMPTION AND COMPOUND INTAKE: Water consumption was slightly low in the
1.8% dose group.

HEMATOLOGY:
- A significant decrease in hematocrit and hemoglobin, a significant increase in the erythrocyte membrane resistance was observed among female animals in the 1.8% dose groups.
- Male and female rats in 1.8% dose group showed increase in white blood cell count, increase in neutrophil fraction out of the white blood cells and a decrease in lymphocytes.

CLINICAL BIOCHEMISTRY: There was dose dependent increase in ALP levels (at 0.6 and 1.8% dose groups) and decrease in total protein (at 1.8% dose group)

URINALYSIS: No effects were observed.

ORGAN WEIGHT:
- In highest concentration group (1.8%), significant increase in relative weights of liver, suprarenal gland, testis, brain and caecum were observed in males and relative weights of heart, thyroid, liver, uterus, brain and caecum of females.In 0.6% dose group, there was significant increase in caecum relative weights in males and hypophysis in females.

In 0.2% dose group, caecum relative weights were significantly increased in males and kidney relative weights were significantly decreased in females.

GROSS PATHOLOGY
- The color of the liver in 2 males and 5 females from the 1.8% dose group was lighter than normal and seemed to have a yellowish white color mixed to it.
- In addition, lung tumors were seen in one female rat from the 0.2% dose group.

- Autopsy of one male rat from the LAS-1.8% dose group that died during the study had significant s welling of the abdomen. This was due to the swelling of the stomach and small intestines. The contents from the stomach to the first part of the jejunum was a blackish purple liquid. From there on, the ileum was filled with a semi-transparent, viscous, gelatinous substance.

HISTOPATHOLOGY

The main findings were concerned with kidneys, liver and intestinal tract. The histological findings concerning the kidneys had differences in the extent and frequency of changes between the dosage and between male and female rats.

Kidneys:
- Chronic appearance of glomeruli indicating moderate level of atrophy (male/females of 0.6% and 1.8% treated groups)
- Glomeruli contributed to overall swelling through dilation of vessel lumen and interstitial swelling (significant in male/female rats of 0.2 and 0.07% dose groups. They were not very significant in the 1.8 and 0.6% dose groups)
- The glomerular interstitum had thawn mildly but chronically (present in control group but was significant for male/female rats in 1.8 and 0.6% groups)
- Cloudy swelling/vacuolar degeneration of proximal renal tubular epithelial cells were seen (mildly present in control group and slightly significant in LAS treated groups).
- Small, yellowish brown blobs the size of nuclei was observed in the proximal renal tubular epithelial cells (significantly in male/females of 1.8% dose group)
- Blue lead color exhibited on the lumen of the Henle loop through H.E. staining, while substances giving a strong positive PAS response were forming layers in circular forms or were present in small pieces
-Lumen dilation of the paroximal renal tube and flattening of epithelial cells (significantly in males of 0.2% dose group, no effect observed in females and control groups)
- The renal tubule had collapsed (significantly in male/females of 1.8% dose group)
- Swelling of surrounding connective tissue was observed in the interstitium (observed in control group and significantly in males of LAS-0.2% dose group and both male and female rats of 0.6% dose group)

- Various changes to the kidneys as seen in 0.2% to 0.07% dose groups differ in their extent and frequency of incidence, but these changes were also seen in the control group, so they were not considered changes that lead to permanent damages like collapse of renal tubule or cyst formation.

Liver: Disappearance of basophilic substances in the hepatocytes and eosin thickening of hepatocytes were observed mildly in male and female rats of the 1.8% dose group.

Intestinal tract: Decrease in height of epithelial cells in the colonic mucosa and disappearance of vacuoles from goblet cells were observed mildly in female rats of the 1.8 and 0.6% dose groups.

Further details on are provided in "Any other information on results inc.tables"

Effect levels

Key result

Dose descriptor:

NOAEL

Effect level:

40 mg/kg bw/day (nominal)

Based on:

act. ingr.

Sex:

male/female

Basis for effect level:

clinical biochemistry

gross pathology

haematology

histopathology: non-neoplastic

organ weights and organ / body weight ratios

Remarks on result:

other: Suppression of weight gain, increase in appendix weight, alteration in hematological and clinical biochemistry parameters, and/ or tissue damages to the large intestines, liver and kidneys in dose levels at ≥ 115 mg/kg bw/day (0.2% diet).

Target system / organ toxicity

Key result

Critical effects observed:

yes

Lowest effective dose / conc.:

115 mg/kg bw/day (nominal)

System:

urinary

Organ:

kidney

Treatment related:

yes

Dose response relationship:

yes

Relevant for humans:

yes

Any other information on results incl. tables

Table 1:  Body weight gain, food, detergent, water intake and feces excretion in males (Yoneyama et al., 1972)

Group	No. of Rats	Initial body wt (g) (mean±S.E)	Final body wt (g) (mean±S.E)	Body weight gain (1) (%) (mean±S.E)	Food (g/rat/day)	Detergent		water (g/rat/day)	Feces (g/rat/day)
						mg/rat/day	mg/kg/day
Control	10	85±0.871	387±9.509	452.1±9.841	14.17	-	-	16.9	2.31
1.80%	10-9	85.2±0.553	290.00±5.206	341.09±5.136**	9.84	177.12	814.8	13.29	1.78
0.60%	10	86.40±1.543	365.80±8.517	426.23±8.893	13.84	83.04	322.3	16.28	2.23
0.20%	10	84.50±0.957	383.40±6.715	453.92±7.603	13.7	27.4	101.5	18.18	2.28
0.07%	10	80.9±1.923	387.10±5.930	480.74±13.113	14.48	10.14	37.2	16.91	2.24

Table 2:  Body weight gain, food, detergent, water intake and feces excretion in females (Yoneyama et al., 1972)

Group	No. of Rats	Initial body wt (g) (mean±S.E)	Final body wt (g) (mean±S.E)	Body weight gain (%) (1) (mean±S.E)	Food (g/rat/day)	Detergent		water (g/rat/day)	Feces (g/rat/day)
						mg/rat/day	mg/kg/day
Control	10	71.80±0.512	202.50±2.325	282.24±4.422	9.35	-	-	17.48	1.53
1.80%	10	74.30±0.943	158.80±3.411	214.02±5.288**	7.01	126.18	913	12.14	1.23
0.60%	10	74.10±0.673	195.80±4.149	264.25±5.207*	9.04	54.24	332.2	13.99	1.53
0.20%	10	74.30±1.116	201.10±3.334	271.05±5.314	8.9	17.8	108.7	17.69	1.39
0.07%	10	76.00±2.683	203.20±7.172	269.35±10.255	9.35	6.55	39.4	19.56	1.63

where (1) refers to Final BW/Initial BW-1) into100

*P<0.05

**P<0.01

Table 3 : Hematology-I in males and females rats (Yoneyama et al., 1972)

Group	No. of Rats	RBC*10^4/mm^3	Ht (%)	Hb (g/dl)	MCH (rr)	MCV (µ8)	MCC%
Male
Control	10	910±13	53.1±0.4	19.8±0.3	21.8±0.4	58.4±1.0	37.4±0.6
1.80%	9	924±27^1)	54.6±0.4	20.6±0.5	22.4±0.8	59.3±1.3	37.7±0.8
0.60%	10	890±14	54.0±0.4	19.8±0.1	22.3±0.3	60.7±0.9	36.7±0.1
0.20%	10	914±25	53.3±0.3	19.5±0.1	21.5±0.6	58.6±1.6	36.7±0.2
0.07%	10	925±20	53.3±0.4	19.6±0.1	21.2±0.5	57.8±1.2	36.7±0.2
Female
Control	10	838±22	52.1±0.5	19.9±0.5	23.9±0.9	62.5±1.6	38.2±1.0
1.80%	10	785±14	49.5±0.3**	18.0±0.1**	23.0±0.4	63.2±0.9	36.4±0.1
0.60%	10	795±17	50.9±0.3	19.0±0.1	24.0±0.5	64.2±1.1	37.4±0.2
0.20%	10	778±16	51.1±0.2	19.1±0.1	24.7±0.4	65.9±1.3	37.5±0.2
0.07%	10	806±18	50.6±0.4	18.9±0.1	23.6±0.6	63.0±1.5	37.4±0.3

Table 4 : Hematology-II in males and females rats (Yoneyama et al., 1972)

Group	No. of Rats	Thrombocyte*10^4(A)	Osmotic fragility (A)	Whole blood clotting time (min)
				Beginning	Ending
Male
Control	10	70.5±9 (8)	213.6±9.4 (9)	1.06±0.13	2.12±0.26
1.80%	9	70.2±4.9 (8)	199.9±6.6 (9)	1.14±0.11	2.32±0.27
0.60%	10	71.6±8.4	211.4±7.9	1.42±0.23	2.71±0.29
0.20%	10	64.4±5.5(9)	228.1±6.5(9)	1.34±0.19	2.50±0.35
0.07%	10	80.1±13.2	225.1±10.5	1.40±0.19	2.52±0.33
Female
Control	10	52.0±3.8 (9)	240.4±7.5 (8)	1.29±0.29	2.95±0.30
1.80%	10	58.4±4.5 (9)	181.8±6.5(8)*	1.37±0.14	2.57±0.19
0.60%	10	54.8±3.3 (9)	226.0±5.9 (9)	1.24±0.17	2.81±0.22
0.20%	10	59.8±5.9 (9)	245.4±9.3(9)	1.26±0.19	2.92±0.22
0.07%	10	63.3±5.3 (8)	248.4±10.2	1.22±0.21	3.30±0.41

*P<0.05

**P<0.01

(A): No of rats

¹⁾ (mean±SE)

Table 5: Percent differential of Leucocytes (Yoneyama et al., 1972)

Group	No. of Rats	Neutro	Lympho	Mono	Eosino
Male
Control	10	9.8±0. 88 (1)	89.08±0.954	0.28±0.102	0.83±0.203
1.80%	9	12.09±0.698*	87.22±0.624	0.09±0.061	0.58±0.164
0.60%	10	10.77±1.017	87.82±0.860	0.26±0.094	0.60±0.158
0.20%	10	10.34±0.521	88.22±0.625	0.40±0.102	0.62±0.128
0.07%	10	9.41±1.074	89.35 ± 0.998	0.50± 0.088	0.73±0.125
Female
Control	10	8.24±0.56	90.85±0.685	0.18±0.073	0.72±0.150
1.80%	10	8.67±0.747	90.61 ± 0.762	0.28±0.076	0.42±0.108
0.60%	10	10.21±1 .054	88.49±1.065	0.32±0.071	0.96±0.143
0.20%	10	8.75±0.754	90.15±0.781	0.38±0.096	0.70±0.146
0.07%	10	8.65±0.752	90.39 ± 0.771	0.40±0.099	0.54±0.108

Table 6: Counts differential of leucocytes.10²/mm³ (Yoneyama et al., 1972)

Group	No. of Rats	WBC	Neutro	Lympho	Mono	Eosino
Male
Control	10	88.12±3.686 (1)	8.49±0.678	78.65±3.779	0.24±0.087	0.67±0.148
1.80%	9	96.16±5.726	11.78±1.113	83.76±4.805	0.08 ±0.057	0.54±0.142
0.60%	10	91.47±5.430	9.96±1.154	80.27±4.686	0.25±0.088	0.58±0.142
0.20%	10	90.55±6.435	9.61±1.166	79.64±5.196	0.36 ±0.089	0.60±0.155
0.07%	10	96.15±3.329	9.08±1.077	85.85±2.913	0.50±0.093	0.71±0.135
Female
Control	10	92.00±3.802	7.59±0.579	83.57±3.517	0.16±0.066	0.67±0.139
1.80%	10	109.30±3.831**	9.34±0.664	95.03 ± 7.208	0.31 ± 0.085	0.47±0.120
0.60%	10	87.90±3.631	8.87±0.847	77.89±3.679	0.26±0.058	0.85±0.133
0.20%	10	83.15±4.350	7.38±0.637	76.67 ± 5.063	0.32±0.078	0.61 ± 0.138
0.07%	10	94.40±5.110	8.36±1.176	85.13±4.180	0.37±0.091	0.51 ± 0.101

*P<0.05

**P<0.01

(A): No of rats

¹⁾(mean±SE)

Table 7: Serum Chemistry (Yoneyama et al., 1972)

Group	No. of Rats	ALP K-A Unit/dl	LAP G-R Unit	GOT karmen unit/ml	UN mg/dl	A/G ratio	Total protein g/dl	Bililubin
								(Indirect) mg/dl	(Direct) mg/dl	Total mg/dl
Male
Control	10	24.07±1.109^1)	185.81± 9.586	132.75± 6.357	16.92±1.102	1.42±0.082	7.36±0.071	0.20±0.050	0.11±0.036	0.29±0.044
1.80%	9	44.96±4.01**	179.80± 9.74	139.12± 9.397	16.30±0.634	1.59±0.129	6.55±0.191**	0.40±0.250	0.12±0.027	0.48±0.268
0.60%	10	29.22±1.093**	180.54± 7.897	133.50± 5.559	15.76±0.611	1.46±0.096	7.20±0.084	0.30±0.201	0.32±0.211	0.56±0.415
0.20%	10	24.34±1.173	181.23± 9.637	140.25±14.732	13.04±1.224*	1.47±0.091	7.20±0.059	0.41±0.231	0.31±0.188	0.68±0.422
0.07%	10	25.48±0.816	178.99± 9.4271	144.60±11.501	15.36±1.221	1.38±0.082	7.26±0.118	0.35±0.201	0.34±0.197	0.64±0.401
Female
Control	10	24.41±1.380	198.54± 8.899	156.30±12.716	17.52±1.733	1.53±0.062	7.80±0.122	0.12±0.050	0.14±0.054	0.23±0.054
1.80%	10	61.41±6.124**	208.72± 9.097	154.60±8.297	19.00±1.043	1.76±0.121	6.42±0.127**	0.12±0.025 (9)	0.12±0.033 (9)	0.25±0.054 (9)
0.60%	10	31.80±1.599**	187.58± 10.642	139.50±3.024	17.15±2.020	1.70±0.097**	7.22±0.186*	0.14±0.036	0.12±0.038	0.27±0.062
0.20%	10	27.23±1.049	193.65± 7.686	146.55±9.144	14.14±1.404	1.48±0.164	7.38±0.184	0.20±0.044 (9)	0.11±0.023 (9)	0.31±0.052 (9)
0.07%	10	25.77±1.237	192.23± 8.056	128.25±5.432	15.09±1.245	1.72±0.067	7.68±0.163	0.15±0.059 (9)	0.10±0.018 (9)	0.24±0.06 (9)

where:

*P<0.05

**P<0.01

(A): No of rats

¹⁾(mean±SE)

Table 8 : Urinalysis in female and male rats (Yoneyama et al., 1972)

Group	No. of Rats	pH				Protein					Glucose		Keton			Blood			Bilirubin		Urobilin
		8	7	6	5	-	±	+	++	+++	-	+	-	±	+	-	±	+	-	+	0.1*	1
Male
Control	10	1	4	5	-	-	-	7	2	1	10	-	7	3	-	10	-	-	10	-	10	-
1.80%	9	-	-	9	-	-	-	2	3	4	9	-	2	7	-	9	-	-	9	-	9	-
0.60%	10	-	5	5	-	-	2	5	2	1	10	-	5	5	-	10	-	-	10	-	10	-
0.20%	10	-	2	8	-	-	4	3	3		10	-	6	4	-	10	-	-	10	-	10	-
0.07%	10	-	4	6	-	1	-	6	1	2	10	-	8	2	-	10	-	-	10	-	10	-
Female
Control	10	2	2	6	-	2	2	5	1	-	10	-	5	5	-	10	-	-	10	-	10	-
1.80%	10	1	-	9	-	3	3	4	-	-	10	-	4	5	-	10	-	-	10	-	9	-
0.60%	10	-	5	5	-	2	3	2	3	-	10	-	7	3	-	10	-	-	10	-	10	-
0.20%	10	-	3	7	-	2	2	5	1	-	10	-	4	6	-	10	-	-	10	-	10	-
0.07%	10	-	3	7	-	3	3	4	-	-	10	-	7	3	-	10	-	-	10	-	9	-

*Ehrlich units

Table 9: Organ Weights (Male-1) (Yoneyama et al., 1972)

Group	No. of rats	Heart [Actual weight (mg)/weight (mg)/100 g]	Spleen [Actual weight (mg)/weight (mg)/100 g]	Thymus [Actual weight (mg)/weight (mg)/100 g]	Liver [Actual weight (g)/weight (g)/100 g]	Lung [Actual weight (mg)/weight (mg)/100 g]	Kidneys [Actual weight (mg)/weight (mg)/100 g]		Hypophysis [Actual weight (mg)/weight (mg)/100 g]
							R	L
Control	10	930±13/241± 3^1)	614±14/158±2	129±5.7/33±0.9	12.0±0.38/3.1±0.04	1211±44/314±13	1105±33/285± 6	1126±33/219± 6	10.0±0.73/2.6±0.21
1.80%	9	694±11**/239±3	460±11**/158±2	102±6.3**/35±1.7	10.3±0.28**/3.5±0.08**	971±35**/324±13	836±19**/288±3	852±22**/294±5	8.8±0.42/3.0±0.15
0.60%	10	878±18*/235± 4	572±15*/153± 4	122±8.8*/32± 2.3	12.0±0.31/3.2±0.07	1080±21/289±6	1045±31/280±9	1049±34/281±9	9.4±0.30/2.5±0.08
0.20%	10	918±16/239± 2	644±27/168±7	130±9.0/33±2.1	12.3±0.42/3.2±0.12	1253±63/326±14	1083±34/282±7	1071±28/279±4	8.9±0.48/2.3±0.13
0.07%	10	932±16/240± 2	641±12/165± 2	129±10.8/33± 2.5	12.0±0.31/3.1±0.05	1285±47/332±12	1074±25/277±5	1083±52/279±12	9.3±0.50/2.4±0.13

Table 10: Organ Weights (Male-2) (Yoneyama et al., 1972)

Group	No. of rats	Thyroid glands [Actual weight (mg)/weight (mg)/100 g]	Supranal gland [Actual weight (mg)/weight (mg)/100 g]		Testis [Actual weight (mg)/weight (mg)/100 g]		Prostate glands [Actual weight (mg)/weight (mg)/100 g]	Brain [Actual weight (mg)/weight (mg)/100 g]	Caecum [Actual weight (mg)/weight (mg)/100 g]
			R	L	R	L
Control	10	14.4±0.421^1)/3.7±0.16	18.0±0.61/4.6±0.18	19.3±0.44/5.0±0.17	1592±20/412± 7	1622±21/420± 6	252±16/65±4.8	1983±12/515±11	837±34/216± 8
1.80%	9	11.5±0.76**/3.9±0.24	15.5±0.44**/5.3±0.17*	17.1±0.65*/5.9±0.27*	1513±23*/522±7**	1501±36**/518±13**	231±21/79±7.0	1960±22/677±11 **	1484±30**/512±12**
0.60%	10	13.2±0.69/3.5±0.22	17.5±0.76/4.6±0.20	18.2±0.69/4.8±0.19	1576±27/422± 11	1612±36/432±11	231±21/61±5.7	1969±18*/528±12	939±41/251±11*
0.20%	10	13.3±0.57/3.4±0.11	17.3±0.61/4.5±0.17	18.6±0.84/4.8±0.23	1564±22/408± 5	1621±20/423± 4	298±21/78±5.9	2020±11/528±8	971±35*/253 ± 7**
0.07%	10	13.6±0.54/3.5±0.13	16.8±0.85/4.3± 0.22	18.7±0.44/4.8± 0.11	1643±14/425± 6	1676±15/434± 7	318±21*/82±6.1	1918±106/494±25	938±38/242± 9

where,

*P<0.05

**P<0.01

(A): No of rats

¹⁾(mean±SE)

Table 11: Organ Weights (Female-1) (Yoneyama et al., 1972)

Group	No. of rats	Heart [Actual weight (mg)/weight (mg)/100 g]	Spleen [Actual weight (mg)/weight (mg)/100 g]	Thymus [Actual weight (mg)/weight (mg)/100 g]	Liver [Actual weight (g)/weight (g)/100 g]	Lung [Actual weight (mg)/weight (mg)/100 g]	Kidneys [Actual weight (mg)/weight (mg)/100 g]		Hypophysis [Actual weight (mg)/weight (mg)/100 g]
							R	L
Control	10	596±100^1)/265± 6	405± 5/200±4	134±7.7/66±3.9	6.5±0.12/3.2±0.08	822±19/409±11	720±22/ 359±12	738±18/395±11	13.9±0.76/6.8±0.39
1.80%	10	426± 11**/269±7*	294± 10**/185± 6	102±6.8**/64±4.1	6.3±0.24/3.9±0.12**	673±20**/425±16	546±16**/345±13	563±24**/357±21	12.3±1.08/7.7±.63
0.60%	10	559± 11*/286± 5	387 ± 7/198±5	125±5.8/63±2.2	6.5±0.22/3.3±0.10	793±14/406±8	664±16/340± 9	715±32/ 365±14	16.2±0.94/8.3±0.46*
0.20%	10	574±13/258± 5	405 ±8/201 ± 4	131±4.6/65±2.6	6.5±0.18/3.2±0.07	802±26/398±10	654 ±20*/325± 7	665±28*/330±11*	15.0±1.10/7.4±0.53
0.07%	10	596±16/294± 3	408±13/201± 4	132±2.8/65±2.3	6.7±0.30/3.3±0.05	801±19/396±11	717±23/355±13	738±33/364±13	16.3±0.94/8.0±0.40

Table 12: Organ Weights (Female-2) (Yoneyama et al., 1972)

Group	No. of rats	Thyroid glands [Actual weight (mg)/weight (mg)/100 g]	Supranal gland [Actual weight (mg)/weight (mg)/100 g]		Ovary [Actual weight (mg)/weight (mg)/100 g]		Uterus [Actual weight (mg)/weight (mg)/100 g]	Brain [Actual weight (mg)/weight (mg)/100 g]	Caecum [Actual weight (mg)/weight (mg)/100 g]
			R	L	R	L
Control	10	11.2±0.321^1)/ 5.5±0.17	21.9±0.99/10.8±0.64	23.2±1.24/11.4±0.94	23.3±1.0/11.5±0.5	24.4±1.1/12.0±0.5	638±27/315±13	1832±16/906±14	681±18/337±11
1.80%	10	10.9±0.73/6.9±0.53*	15.6±0.81**/9.8±0.52	16.8±0.84**/10.6±0.55	17.6±1.4**/11.1±0.9	18.2±1.8**/11.5±1.1	292±18**/185±12**	1780±15*/1126±28	929 ± 26**/586±14**
0.60%	10	13.9±1.30/7.1±0.70	22.9±0.56/11.7±0.36	24.0±0.61/12.3±0.44	22.8±1.2/11.7±0.7	24.5±1.0/12.6±0.6	618±32/315±15	1840±14/943±20	697±21/356± 7
0.20%	10	12.2±0.96/6.0±0.46	21.7±1.11/10.7±0.49	22.2±0.92/11.0±0.43	23.6±1.0/11.7±0.4	23.3±1.2/11.5±0.5	594±31/296±17	1837±18/915±13	688±26/343±14
0.07%	10	12.9±0.93/6.4±0.55	22.6±0.95/11.1±0.35	24.1±0.98/11.8±0.33	23.0±1.9/11.2±0.6	24.6±1.5/12.0±0.5	592±18/292± 8	1827±18/908±30	753±25*/373±13

where,

*P<0.05

**P<0.01

(A): No of rats

¹⁾(mean±SE)

Table 13: Summary of microscopic  changes in kindey (Yoneyama et al., 1972)

Microscopic findings	#	Male					Female
		C	LAS				C	LAS
			0.07%	0.2%	0.6%	1.8%		0.07%	0.2%	0.6%	1.8%
Swelling in glomerulus	S E M O S L	0 2 (10) 2	0 5 (10) 4	1 6 (10) 3	0 7 (10) 2	0 5 (9) 4	0 2 (10) 3	1 5 (10) 4	1 5 (10) 4	0 7 (10) 3	0 5 (10) 5
Histolysis in glomerulus	S E M O S L	0 2 2	0 7 2	1 5 4	1 5 4	1 5 3	0 3 1	0 5 5	0 9 4	1 6 3	1 5 4
Atrophy in glomerulus	S E M O S L	0 1 1	0 5 5	1 6 2	2 7 1	3 6 0	0 1 1	0 4 5	0 6 3	1 7 2	4 5 1
Cloudy swelling in epithelium of proximal tubules	S E M O S L	0 0 4	0 1 5	0 6 2	0 8 2	0 7 2	0 0 3	0 0 4	0 3 3	0 3 3	0 4 5
Vacuolation in epithelium of proximal tubules	S E M O S L	0 0 4	0 6 4	0 3 7	0 4 6	0 5 4	0 1 5	0 7 3	0 6 4	0 7 3	0 7 3
Lipofuscin in epithelium of proximal tubules	S E M O S L	0 0 0	0 0 0	0 0 5	0 0 5	2 7 0	0 0 0	0 0 0	0 0 1	0 0 4	5 5 0
PAS-positive materies in loop of Henle	S E M O S L						0 0 4	0 3 7	0 3 6	0 3 6	0 0 3
Dilatation in lumina of distal tubules	S E M O S L	0 0 2	0 4 6	3 3 4	2 4 4	0 5 4	0 0 0	0 0 5	0 3 3	0 5 5	0 3 5
Decrease in cell height of epithelium in distal tubules	S E M O S L	0 0 1	0 0 6	2 3 5	2 3 4	0 4 5	0 0 0	0 0 3	0 0 5	0 3 5	0 3 5
Destruction in urinary tubules	S E M O S L	0 0 0	0 0 0	0 0 2	0 2 3	0 3 4	0 0 0	0 0 2	0 0 3	0 0 5	0 4 5
edema in stroma	S E M O S L	0 1 2	1 5 4	1 6 3	1 3 5	0 4 5	0 1 1	1 4 5	1 5 4	1 6 3	0 2 7
Swelling in perivascular connective tissues	S E M O S L	0 1 1	1 4 4	1 3 6	0 5 5	0 3 4	0 1 1	0 4 5	0 5 5	0 7 3	0 1 6
Proliferation in interstitial connective tissues	S E M O S L	0 0 1	0 0 5	0 0 5	0 1 4	0 2 6	0 0 2	0 0 2	0 0 5	0 0 6	0 0 6

where, SE: Severe

SL: Slight

MO: Moderate

(A) No. of rats

Table 14:  Summary of microscopic changes in liver and large intestine (Yoneyama et al., 1972)

Microscopic findings	#	Male					Female
		C	LAS				C	LAS
			0.07%	0.2%	0.6%	1.8%		0.07%	0.2%	0.6%	1.8%
Decrease in basoplam in liver cells	S E M O S L	0 0 (10) 0	0 0 (10) 0	0 0 (10) 1	0 0 (10) 1	0 0 (9) 6	0 0 (10) 0	0 0 (10) 0	0 0 (10) 0	0 0 (10) 3	0 1 (10) 6
Increase in eosinophilia in liver cells	S E M O S L	0 0 1	0 0 0	0 0 0	0 0 0	0 0 6	0 0 1	0 0 0	0 0 0	0 0 2	0 2 7
Swelling in liver cells	S E M O S L	0 0 0	0 0 0	0 0 0	0 3 3	0 3 3	0 0 0	0 0 0	0 1 3	0 1 4	0 2 6
Rarefaction in cytoplasm of liver cells	S E M O S L	0 0 3	0 0 8	0 1 6	0 1 6	0 1 6	0 0 2	0 0 6	0 1 7	0 1 7	0 1 7
Decrease in cell height of epithelium in large instestine	S E M O S L	0 0 0	0 0 0	0 0 2	0 0 4	0 0 5	0 0 0	0 0 0	0 0 3	0 1 4	0 1 4

where, SE: Severe

SL: Slight

MO: Moderate

(A) No. of rats

Applicant's summary and conclusion

Conclusions:

Administration of linear alkylbenzene sulfonic acid sodium salt (LAS) to Wistar SLC rats at dose levels of 0, 0.07, 0.2, 0.6 and 1.8% (equivalent to 0, 40, 115, 340, 1030 mg/kg bw/day) for 26 weeks revealed an NOAEL of 0.07% (40 mg/kg bw/day), based on tissue damage in caecum, kidney and liver, hematological and/or clinical chemistry parameters changes at higher dose levels (115 mg/kg bw/day and above).

Executive summary:

A sub-chronic repeated dose toxicity study was conducted with C10-14 LAS, sodium salt in Wistar SLC rats. The test substance was administered to groups of 10 male and female rats/dose in the diet for 26 weeks at doses of 0, 40, 115, 340 and 1030 mg/kg/day. Clinical observations, water consumption, food consumption and body weights were recorded weekly. At the end of study, haematological, clinical chemistry and urine parameters were analysed. Gross pathological examinations were observed after necropsy and the major organs were subjected to histopathological examinations. Significant diarrhoea, suppressed growth, increased caecal weight and degeneration of renal tubes were observed in the highest dose group. Similar but less severe signs were seen in other doses with the exception of the lowest dose of 40 mg/kg/day, which showed no adverse effects related to exposure to the LAS. Under the study conditions, systemic toxicity NOAEL was determined to be 40 mg/kg/day, based on tissue damage in caecum, kidney and liver, haematological as well as clinical chemistry parameters changes at higher dose levels (Yoneyama, 1972).