Benzenesulfonic acid, 4-C10-13-sec-alkyl derivs.

Repeated dose toxicity: oral

Administrative data

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

1976

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

Reference

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

1976

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

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

Reason / purpose for cross-reference:

reference to same study

Qualifier:

no guideline followed

Principles of method if other than guideline:

The purpose of this study is to determine the sub chronic toxicity of LAS in SLC-ICR mice, focusing on the liver and kidneys. Male and female mice were maintained on either test diets (0 and 0.6%) or drinking water (0, 0.07, 0.2 and 0.6%) for 9 months. Mice in 1.8% dose group of both feeding and drinking water study exhibited severe weight loss so LAS administration was stopped after 2 weeks. Mortality, clinical observations and body weights were recorded during the study. All the surviving animals were humanely euthanized at end of 9 months and gross necropsy, enzyme tests on the liver and kidneys were performed and organs weights were measured. No other parameters (histopathology, clinical chemistry and haematological parameters) were evaluated in the study.

GLP compliance:

no

Remarks:

predates GLP

Limit test:

no

Species:

mouse

Strain:

other: SLC-ICR mice

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: 4-weeks old
- Weight at study initiation: 24 - 31 g (male mice), 20 - 25 g (female mice)
- Fasting period before study: No
- Housing: 5 animals were housed per cage.
- Diet: CE-2 food (from CLEA Japan); ad libitum
- Water: ad libitum
- Acclimation period: 1 week before the start of treatment

ENVIRONMENTAL CONDITIONS
- Temperature: 25 ± 1°C
- Humidity: 50 - 60%
- Air changes: Not reported
- Photoperiod: 12 hours dark /12 hours light

IN-LIFE DATES: Not reported

Route of administration:

other: oral: drinking water and feed

Details on route of administration:

LAS was administered to animals by mixing 0.6 and 1.8% in CE-2 food (CLEA Japan) and dissolving to 0.07, 0.2, 0.6 and 1.8% in drinking water.

Vehicle:

other: Test substance was administered either in diet or drinking water.

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

9 months

Frequency of treatment:

Continuous in diet or drinking water (ad libitum)

Dose / conc.:

780 mg/kg bw/day (nominal)

Remarks:

in diet (corresponding to 0.6% dose level)

Dose / conc.:

2 340 mg/kg bw/day (nominal)

Remarks:

in diet (corresponding to 1.8% dose level) was also included, however due to severe weight loss so LAS administration was stopped after 2 weeks.

Dose / conc.:

133 mg/kg bw/day (nominal)

Remarks:

in drinking water (corresponding to 0.07% dose level)

Dose / conc.:

380 mg/kg bw/day (nominal)

Remarks:

in drinking water (corresponding to 0.2% dose level)

Dose / conc.:

1 140 mg/kg bw/day (nominal)

Remarks:

in drinking water (corresponding to 0.6% dose level)

Dose / conc.:

3 420 mg/kg bw/day (nominal)

Remarks:

in drinking water (corresponding to 1.8% dose level) was also included, however due to severe weight loss so LAS administration was stopped after 2 weeks.

No. of animals per sex per dose:

Feeding study (mixed in diet): 9 males in 0.6% fed group and 8 males in control group; 8 females in control as well as 0.6% fed group
Drinking water study: 9 animals each in test substance treatment groups and 8 animals in control group

Control animals:

yes, concurrent vehicle

yes, plain diet

Positive control:

No

Observations and examinations performed and frequency:

CLINICAL OBSERVATIONS: Yes
- Time schedule: Weekly

BODY WEIGHT: Yes
- Time schedule for examinations: Weekly

FOOD CONSUMPTION AND COMPOUND INTAKE:
- Food consumption: Yes
- Compound intake: Yes
- Time schedule for examinations: Weekly

FOOD EFFICIENCY: No

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

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: No

CLINICAL CHEMISTRY: No

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No

IMMUNOLOGY: No

OTHER:
- Liver enzyme tests: Glucose 6-phosphatase (G6Pase), lactase dehydrogenase (LDH), GPT and GOT
- Kidney enzyme tests: G6Pase, LDH, GPT, GOT, ALP, acid phosphatase (ACP), Na, K-ATPase, and malate dehydrogenase (MDH)

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
Gross findings were obtained after euthanizing animals and organs were removed for organ weight measurements.

ORGAN WEIGHTS: Heart, lungs, liver, spleen, kidneys, testes and uterus

HISTOPATHOLOGY: No

Clinical signs:

no effects observed

Mortality:

mortality observed, non-treatment-related

Description (incidence):

The number of males in 0.2% dose group (drinking water study) decrease from 9 to 5 animals due to an accident with housing management during the experiment.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Dietary study: There was a significant increase in body weight compared to controls in males of 0.6% dose group.

Drinking water study: There was significant inhibition of weight increase in both males and females of 0.6% dose group consuming LAS water compared to controls, while there was a significant increase in body weight compared to controls in males of 0.07% dose group consuming LAS water.

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

Individual daily food consumption was higher in all treatment groups compared to controls.

Food efficiency:

not examined

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

effects observed, treatment-related

Description (incidence and severity):

Dietary study: Water consumption was reduced in females and increased in males of 0.6% dose group (dietary study)

Drinking water study: Water consumption was reduced in males of 0.6% dose group and in females of 0.2% dose group and increased in males of 0.07 and 0.2% dose groups and females of 0.07% dose groups.

Ophthalmological findings:

not examined

Haematological findings:

not examined

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

not examined

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):

Dietary study: In males of 0.6% dose group, the absolute liver weight was significantly increased, in males of 0.6% dose group (dietary study), the relative liver weight was significantly increased. In females, the relative liver weight was significantly increased in all treatment groups.

Drinking water study: In males of 0.07% dose group, the absolute liver weight was significantly increased, in males of 0.2 and 0.6% dose groups, the relative liver weight was significantly increased, and in 0.07% males the relative liver weight was increased. Relative lung, heart and kidney weights were also increased in male mice consuming 0.07% LAS water. In females, the relative liver weight was significantly increased in all treatment groups. Relative spleen, heart and kidney weights were also increased in female mice consuming 0.6% LAS water.

Gross pathological findings:

not specified

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

not examined

Histopathological findings: neoplastic:

not examined

Other effects:

effects observed, treatment-related

Description (incidence and severity):

Liver enzyme tests:
Dietary study: Only there was reductions in LDH activity in male mice of 0.6% group
Drinking water study: GOT activity in male mice of 0.6% group consuming LAS water were significant. GOT activity in female mice of 0.2% group was also significantly reduced.

Renal enzyme tests: G6Pase was significantly reduced in males and female mice of 0.6% group consuming LAS water compared to controls, and also reduced in other treatment groups.

Dose descriptor:

other: no NOAEL can be determined for dietary study

Remarks:

Dietary study

Remarks on result:

other: Only one dose level for LAS was fed to animals throughout the study.

Dose descriptor:

NOAEL

Remarks:

Drinking water study

Effect level:

133 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

body weight and weight gain

organ weights and organ / body weight ratios

other: Liver and kidney enzyme levels

Remarks on result:

other: This is based on alteration in liver and kidney weights and enzymes levels at higher tested dose levle. As no other parameters (histopathology, clinical chemistry and hematological parameters) were evaluated in the study.

Critical effects observed:

yes

Lowest effective dose / conc.:

380 mg/kg bw/day (nominal)

System:

urinary

Organ:

kidney

Treatment related:

yes

Dose response relationship:

yes

Relevant for humans:

yes

Critical effects observed:

yes

Lowest effective dose / conc.:

380 mg/kg bw/day (nominal)

System:

hepatobiliary

Organ:

liver

Treatment related:

yes

Dose response relationship:

yes

Relevant for humans:

yes

Conclusions:

Administration of LAS to SLC-ICR mice by either test diets (0 and 0.6) or drinking water (0, 0.07, 0.2 and 0.6) for 9 months (focusing on the liver and kidneys) revealed an NOAEL of 0.07% (133 mg/kg bw/day in drinking water), based on observed adverse effects on organ weight and liver and kidney enzymes. Repeated administration of LAS impair renal and liver function.

Executive summary:

The 9 months sub-chronic oral toxicity study of LAS was performed in SLC-ICR mice, focusing on the liver and kidneys.

4 weeks old male and female SLC-ICR mice(obtained from Shizuoka Agricultural Cooperative Association for Laboratory Animals) with body weight range 24 - 31 g (males), 20 - 25 g (females) were used in the study. 5 animals were housed in each cage and maintained under controlled environmental conditions (temperature: Average of25 ± 1°C, humidity:50 - 60%, and 12 hours light /12 hours dark). CE-2 diet (from CLEA Japan) and water were provided ad libitum.

The animals were administered daily with the LAS at following dose levels for 9 months:

Mixed in diet: 0 and 0.6% (equivalent to 0 and 780 mg/kg bw/day); 9 males in 0.6% fed group and 8 males in control group; 8 females in control as well as 0.6% fed group

Dissolved in drinking water: 0, 0.07, 0.2 and 0.6 (equivalent to 0,133, 380 and 1140 mg/kg bw/day); 9 animals each in test substance treatment groups and 8 animals in control group

Mice in 1.8% dose group of both feeding and drinking water study exhibited severe weight loss so LAS administration was stopped after 2 weeks.

Clinical observations, water consumption, food consumption and body weights were recorded weekly. At the end of study, blood was collected for estimation of haematological and clinical chemistry parameters. Gross findings were obtained after euthanizing animals and organs were removed for organ weight measurements. Organ weights for brain, heart, lungs, liver, spleen, kidneys, adrenal gland, testes, uterus, and appendix were recorded. Liver and kidney enzymes were also analysed.

No mortality was observed throughout the study. There was significant inhibition of weight increase in both males and females of 0.6% dose group consuming LAS water compared to controls, while there was a significant increase in body weight compared to controls in males of 0.6% dose group fed with LAS in diet and 0.07% dose group consuming LAS water. Individual daily food consumption was higher in all treatment groups compared to controls. Water consumption was reduced in males of 0.6% dose group (drinking water study) and in females of 0.6% dose group (dietary study) and 0.2% dose group (drinking water study) and increased in males of 0.6% dose group (dietary study), 0.07 and 0.2% dose groups (drinking water study) and female of 0.07% dose groups (drinking water study).

In males of 0.6% dose group (dietary study) and 0.07% dose group (drinking water study), the absolute liver weight was significantly increased, in males of 0.6% dose group (dietary study), 0.2 and 0.6 dose groups (drinking water study), the relative liver weight was significantly increased, and in 0.07% (drinking water) males the relative liver weight was increased. Relative lung, heart and kidney weights were also increased in male mice consuming 0.07% LAS water. In females, the relative liver weight was significantly increased in all treatment groups. Relative spleen, heart and kidney weights were also increased in female mice consuming 0.6% LAS water.

There was reductions in LDH activity in male mice of 0.6% group fed with LAS diet and GOT activity in male mice of 0.6% group consuming LAS water were significant. GOT activity in female mice of 0.2% group (drinking water study) was also significantly reduced. G6Pase was significantly reduced in males and female mice of 0.6% group consuming LAS water compared to controls, and also reduced in other treatment groups.

No NOAEL can be identified for dietary study in mice, as only one dose level for LAS was fed to animals throughout the study.

Administration of LAS to SLC-ICR mice by either test diets (0 and 0.6) or drinking water (0, 0.07, 0.2 and 0.6) for 9 months (focusing on the liver and kidneys) revealed an NOAEL of 0.07% (133 mg/kg bw/day in drinking water), based on observed adverse effects on organ weight and liver and kidney enzymes. Repeated administration of LAS impair renal and liver function.

Data source

Reference

Reference Type:

publication

Title:

Unnamed

Year:

1976

Materials and methods

Principles of method if other than guideline:

The purpose of this study is to determine the sub chronic toxicity of LAS in Wistar JCL rats, focusing on the liver and kidneys. Male and female rats were maintained on either test diets (0, 0.6 and 1.8%) or drinking water (0, 0.07 and 0.2%) for 9 months. 0.6 and 1.8% rats in drinking water study exhibited severe weight loss so LAS administration was stopped after 2 weeks. Mortality, clinical observations and body weights were recorded during the study. All the surviving animals were humanely euthanized at end of 9 months and gross necropsy, haematological, serum biochemical tests, enzyme tests on the liver and kidneys were performed and organs weights were measured. No histopathology was 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:

not specified

Test animals

Species:

rat

Strain:

other: Wistar JCL

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Male and female rats were obtained from Shizuoka Agricultural Cooperative Association for Laboratory Animals
- Age at study initiation: 4-weeks old
- Weight at study initiation: 100 - 124 g (male rats), 82 - 100 g (female rats)
- Fasting period before study: No
- Housing: 5 animals were housed per cage.
- Diet: CE-2 food (from CLEA Japan); ad libitum
- Water: ad libitum
- Acclimation period: 1 week before the start of treatment

ENVIRONMENTAL CONDITIONS
- Temperature: 25 ± 1°C
- Humidity: 50 - 60%
- Air changes: Not reported
- Photoperiod: 12 hours dark /12 hours light

IN-LIFE DATES: Not reported

Administration / exposure

Route of administration:

other: oral: drinking water and feed

Details on route of administration:

LAS was administered to animals by mixing 0.6 and 1.8% in CE-2 food (CLEA Japan) and dissolving to 0.07 and 0.2% in drinking water.

Vehicle:

other: Test substance was administered either in diet or drinking water

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

9 months

Frequency of treatment:

Continuous in diet or drinking water (ad libitum)

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

0.6 other: %

Remarks:

in diet (corresponding to 300 mg/kg bw/day)

Doses / concentrations 2

Dose / conc.:

1.8 other: %

Remarks:

in diet (corresponding to 900 mg/kg bw/day)

Doses / concentrations 3

Dose / conc.:

0.07 other: %

Remarks:

in drinking water (corresponding to 85 mg/kg bw/day)

Doses / concentrations 4

Dose / conc.:

0.2 other: %

Remarks:

in drinking water (corresponding to 145 mg/kg bw/day)

No. of animals per sex per dose:

Feeding study (mixed in diet): 8 animals/sex/dose
Drinking water study: 9 animals/sex/dose

Control animals:

yes, concurrent vehicle

yes, plain diet

Positive control:

No

Examinations

Observations and examinations performed and frequency:

CLINICAL OBSERVATIONS: Yes
- Time schedule: Weekly

BODY WEIGHT: Yes
- Time schedule for examinations: Weekly

FOOD CONSUMPTION AND COMPOUND INTAKE:
- Food consumption: Yes
- Compound intake: Yes
- Time schedule for examinations: Weekly

FOOD EFFICIENCY: No

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

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: After 9 months, blood was collected from each animal after euthanizing.
- Anesthetic used for blood collection: No
- Animals fasted: Not specified
- How many animals: Not specified
- Parameters: White blood cells (WBC), red blood cells (RBC), Haemoglobin (Hgb), Haematocrit (Hct), mean corpuscular volume (MCV) and mean corpuscular haemoglobin concentration (MCHC)

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: After 9 months, blood was collected from each animal after euthanizing.
- Animals fasted: Not specified
- How many animals: Not specified
- Parameters: Glutamate oxaloacetate transaminase (GOT), glutamate pyruvic transaminase (GPT), glucose content, urea nitrogen, total cholesterol, albumin, alkaline phosphatase (ALP) and cholinesterase

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No

IMMUNOLOGY: No

OTHER:
- Liver enzyme tests: Glucose 6-phosphatase (G6Pase), lactase dehydrogenase (LDH) and G6P-DH activity
- Kidney enzyme tests: G6Pase, LDH, GPT, GOT, ALP, acid phosphatase (ACP), Na, K-ATPase, and malate dehydrogenase (MDH)

Sacrifice and pathology:

GROSS PATHOLOGY: Yes (However, no details in study report in mentioned)
Gross findings were obtained after euthanizing animals and organs were removed for organ weight measurements.

ORGAN WEIGHTS: Brain, heart, lungs, liver, spleen, kidneys, adrenal gland, testes, ovary, uterus, and appendix

HISTOPATHOLOGY: No

Results and discussion

Results of examinations

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

Both female and male rats consuming LAS-containing water exhibited a slight redness at the tips of their facial fur, and coarse fur all over their bodies.

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

There was significant decrease in body weight gain in males and females of 1.8% dose group fed with diet and in males of 0.2% dose group consuming LAS contained water.

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

not examined

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

effects observed, treatment-related

Description (incidence and severity):

Increased water consumption was observed in males of 1.8% dose group fed with diet and 0.2% dose group consuming LAS contained water.

Ophthalmological findings:

not examined

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

A significant reduction in WBC was observed in 0.6% (diet) male rats compared to controls, and a significant reduction in MCV and MCH was observed in 1.8% (diet) female rats compared to controls.

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

Except female rats of 0.07% dose group (drinking water), a significant reduction or a reduction in cholesterol was observed in male and female rats of all dose groups compared to controls. GPT was significantly reduced in 0.6% dose group (diet) females, and reduced in females of other treatment groups. GOT was significantly reduced in 1.8% dose group (diet) males, and reduced in both females and males of other treatment groups. Albumin was significantly reduced in 1.8% (diet) males and reduced in males of 0.2% dose group (drinking water) and females of 1.8% dose group (diet). There was significant increase in ALP levels in male and female rats fed with 1.8% LAS-diet and cholinesterase levels in male rats fed with 1.8% LAS-diet.

Urinalysis findings:

not examined

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):

In males of 1.8% (diet), the absolute liver and testes weight was significantly decreased and the relative testes and liver weight was significantly increased and both absolute and relative spleen weight were also significantly reduced.
In females of 1.8% (diet), absolute and relative liver and caecum weight was significantly increased.
There was also significant decrease in absolute and relative heart weight in females of 0.6% (diet) and 0.07% (drinking water) dose groups.

Gross pathological findings:

not specified

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

not examined

Histopathological findings: neoplastic:

not examined

Other effects:

effects observed, treatment-related

Description (incidence and severity):

Liver enzyme tests:
Dietary study: G6Pase activity was reduced in 1.8% dose group males and females, G6P-DH activity was reduced in 0.6 and 1.8% dose group males and females, LDH activity was reduced in 0.6, 1.8% dose group. GOT and GPT activities were clearly reduced in males, while in females GPT activity was reduced only in 1.8% dose group and GOT activity was reduced in 1.8% dose group animals.
Drinking water study: LDH activity was reduced in 0.2% dose group males. GOT and GPT activities were clearly reduced in males and GOT activity was increased in 0.07 and 0.2% dose group animals.

Renal enzyme tests:
Dietary study: A significant difference was also observed in G6Pase activity. In females, G6Pase, Na, K-ATPase, and LDH activity were significantly reduced in 1.8% dose group (diet) animals
Drinking water study: In males, Na, K-ATPase activity was significantly reduced in 0.2% dose group. A significant difference was also observed in G6Pase activity.

Effect levels

1

Dose descriptor:

LOAEL

Effect level:

300 mg/kg bw/day (nominal)

Based on:

act. ingr.

Remarks:

(dietery study)

Sex:

male/female

Basis for effect level:

body weight and weight gain

clinical biochemistry

clinical signs

haematology

organ weights and organ / body weight ratios

water consumption and compound intake

other: Liver and kidney enzyme levels

Remarks on result:

other: Adverse effects were observed at all dose levels

2

Dose descriptor:

NOAEL

Effect level:

85 mg/kg bw/day (nominal)

Based on:

act. ingr.

Remarks:

(drinking water study)

Sex:

male/female

Basis for effect level:

clinical biochemistry

other: Liver and kidney enzyme levels

Remarks on result:

other: Based on significant decreases in the activities of glutamate-oxalate transaminase and lactate dehydrogenase in males and significant decrease in renal Na,K-ATPase in males and females at 145 mg/kg bw/day (0.2% drinking water).

Target system / organ toxicity

1

Critical effects observed:

yes

Lowest effective dose / conc.:

145 mg/kg bw/day (nominal)

System:

urinary

Organ:

kidney

Treatment related:

yes

Dose response relationship:

yes

Relevant for humans:

yes

2

Critical effects observed:

yes

Lowest effective dose / conc.:

145 mg/kg bw/day (nominal)

System:

hepatobiliary

Organ:

liver

Treatment related:

yes

Dose response relationship:

yes

Relevant for humans:

yes

Any other information on results incl. tables

Table 1: Body weight gain, food, water and sample consumption of rats on administration of LAS for 9 months (Yoneyama et. al., 1976)

Sex	Groups	No. of rats	Initial body weight (g)	Final Body weight (g)	Body weight gain (%)	Food (g/rat/day)	Water (g/rat/day)	Sample (g/kg BW/day)
Male	Control	8	113.1± 1.156	421.6 ± 8.542	372.6 ± 5.889	15	23	-
	0.6 F	8	110.6 ± 0.925	413.5 ± 7.356	373.8 ± 6.380	16	23	0.234
	1.8 F	8	109.2 ± 2.218	341.1 ± 8.008	312.5 ± 6.518**	14	33	0.747
	0.07 W	9	110.6 ± 1.572	411.0 ± 5.255	371.9 ± 6.908	17	29	0.051
	0.2 W	9	108.3 ± 1.900	385.1 ± 5.453	356.1 ± 6.346 **	15	33	0.148
Female	Control	8	92.3 ± 1.592	212.6 ± 4.508	230.5 ± 5.420	10	18	-
	0.6 F	8	92.1 ± 1.663	216.1 ± 7.705	234.3 ± 5.753	10	17	0.287
	1.8 F	8	90.6 ± 1.463	184.8 ± 4.278	204.3 ± 5.791 **	9	18	0.969
	0.07 W	9	92.7 ± 1.770	206.0 ± 3.555	222.4 ± 4.478	11	20	0.082
	0.2 W	9	91.6 ± 1.893	209.3 ± 5.383	228.4 ± 3.857	12	18	0.173

Values: mean ± S.E.

*p<0.05

**p<0.01

Table 2: Blood components of rats on administration of LAS for 9 months (Yoneyama et. al., 1976)

Sex	Groups	WBC ( x 10^3)	RBC ( x 10^6)	HGB (g)	HCT (%)	MCV (µ^3)	MCH (µµg)	MCHC (%)
Male	Control	5.5±0.414	5.7 ± 0.142	10.9 ± 0.195	28.0 ± 0.663	49.0 ± 0.189	19.5 ± 0.247	39.5 ± 0.504
	0.6 F	4.2 ± 0.221 *	5.9 ± 0.159	11.2 ± 0.291	29.2 ± 1.059	49.6 ± 0.778	19.4 ± 0.436	38.9 ± 0.851
	1.8 F	4.9 ± 0.461	5.8 ± 0.076	10.7 ± 0.118	27.8 ± 0.561	49.2 ± 0.250	19.1 ± 0.268	38.5 ± 0.499
	0.07 W	4.7 ± 0.209	5.9 ± 0.072	11.3 ± 0.094	29.1 ± 0.317	49.2 ± 0.147	19.1 ± 0.210	38.6 ± 0.447
	0.2 W	4.9 ± 0.304	6.0 ± 0.119	11.3 ± 0.141	29.3 ± 0.566	48.8 ± 0.200	19.1 ± 0.238	38.7 ± 0.493
Female	Control	2.6 ± 0.297	5.6 ± 0.163	11.0 ± 0.243	28.8 ± 0.859	51.7 ± 0.164	20.0 ± 0.364	38.4 ± 0.689
	0.6 F	2.5 ±0.150	5.7 ± 0.265	11.2 ± 0.485	29.3 ± 1.400	51.2 ± 0.313	19.8 ± 0.356	38.5 ± 0.699
	1.8 F	3.7 ± 0.556	5.6 ± 0.294	10.5 ± 0.599	28.2 ± 1.546	50.0 ± 0.378 **	18.9 ± 0.193*	37.5 ± 0.294
	0.07 W	2.4 ± 0.213	6.0 ± 0.302	11.5 ± 0.556	30.5 ± 1.551	51.5 ± 0.176	19.5 ± 0.159	37.7 ± 0.299
	0.2 W	2.7 ± 0.328	5.6 ± 0.100	11.0 ± 0.176	28.9 ± 0.624	52.0 ± 0.463	20.0 ± 0.349	38.3 ± 0.601

Values: mean ± S.E.

*p<0.05

**p<0.01

Table 3: Biochemical values in serum of rats on administration of LAS for 9 months (Yoneyama et. al., 1976)

Sex	Groups	GOT (K.U/mL)	GPT (K.U/mL)	GLU (mg/dL)	UN (mg/dL)	CHO (mg/dL)	ALB (g/dL)	ALP ((KA-U/dL)	ChE (ΔpH)
Male	Control	151.3 ± 7.285	107.5 ± 8.177	163.0 ± 11.488	21.0 ± 0.333	77.5 ± 3.417	4.87 ± 0.055	41.3 ± 1.308	0.166 ± 0.010
	0.6 F	133.2 ± 5.573	102.3 ± 8.485	132.2 ± 4.139*	21.0 ± 0.684	71.6 ± 2.314	47.2 ± 0.075	44.7 ± 2.782	0.187 ± 0.013
	1.8 F	126.7 ± 6.439*	93.0 ± 3.665	144.7 ± 10.099	23.1 ± 0.490**	53.0 ± 1.282**	44.7 ± 0.068**	82.3 ± 3.731**	0.281 ± 0.007 **
	0.07 W	142.8 ± 10.111	108.5 ± 12.381	131.6 ± 3.088*	22.8 ± 0.647*	68.2 ± 2.527*	47.5 ± 0.059	44.0 ± 1.239	0.131 ± 0.015
	0.2 W	143.8 ± 9.041	103.3 ± 9.513	165.1 ± 14.043	21.0 ± 1.462	62.5 ± 2.534**	43.2 ± 0.251	34.4 ± 3.193	0.171 ± 0.008
Female	Control	142.3 ± 14.359	104.2 ± 12.474	135.5 ± 5.246	23.2 ± 0.793	95.8 ± 4.778	47.1 ± 0.265	45.0 ± 5.728	0.963 ± 0.073
	0.6 F	116.1 ± 5.244	68.7 ± 2.670*	126.4 ± 3.657	21.5 ± 0.602	86.7 ± 4.230	4.85 ± 0.095	40.7 ± 1.539	1.056 ± 0.023
	1.8 F	133.6 ± 4.007	88.1 ± 4.362	130.8 ± 3.945	26.7 ± 1.935	67.8 ± 7.472**	42.8 ± 0.097	73.8 ± 11.808*	0.861 ± 0.053
	0.07 W	121.2 ± 7.003	74.2 ± 13.529	127.2 ± 2.035	21.9 ± 0.872	95.0 ± 3.755	49.0 ± 0.187	45.6 ± 12.960	0.881 ± 0.171
	0.2 W	128.5 ± 12.209	85.7 ± 9.362	136.1 ± 10.070	21.7 ± 0.950	77.2 ± 8.826	46.8 ± 0.155	44.0 ± 5.756	1.053 ± 0.042

Values: mean ± S.E.

*p<0.05

**p<0.01

Table 4:  Organ weight of male and female rats on administration of LAS for 9 months (Yoneyama et. al., 1976)

Sex

Groups

Brain

Heart

Lung

Liver

Spleen

Kidney (R)

Kidney (L)

Adrenal (R)

Adrenal (L)

Testis(R)

Testis(L)

Ovary (R)

Ovary (L)

Caecum

Uterus

g

g/100g

g

g/100g

g

g/100g

g

g/100g

g

g/100g

g

g/100g

g

g/100g

g

g/100g

g

g/100g

g

g/100g

g

g/100g

g

g/100g

g

g/100g

g

g/100g

g

g/100g

Male

Control

1.987 ± 0.011

0.463 ± 0.008

1.026 ± 0.018

0.239 ± 0.005

1.263 ± 0.033

0.294 ± 0.008

12.7 ± 0.341

2.9 ± 0.039

0.698 ± 0.014

0.162 ± 0.003

1.162 ± 0.40

0.270 ± 0.008

1.184 ± 0.041

0.275 ± 0.007

18.3 ± 1.117

4.2 ± 0.217

18.1 ± 0.934

4.2 ± 0.225

1.591 ± 0.023

0.370 ± 0.006

1.643 ± 0.027

0.382 ± 0.005

-

-

-

-

1.238 ± 0.068

0.281 ± 0.015

-

-

0.6 F

1.982 ± 0.011

0.473 ± 0.007

0.971 ± 0.022*

0.231 ± 0.004

1.269 ± 0.042

0.302 ± 0.005

12.9 ± 0.310

3.0 ± 0.049

0.682 ± 0.019

0.162 ± 0.002

1.210 ± 0.026

0.290 ± 0.006

1.214 ± 0.032

0.289 ± 0.007

19.6 ± 1.900

4.7 ± 0.435

17.6 ± 1.602

4.2 ± 0.416

1.563 ± 0.019

0.373 ± 0.004

1.585 ± 0.027

0.378 ± 0.007

-

-

-

-

1.303 ± 0.057

0.305 ± 0.010

-

-

1.8 F

1.827 ± 0.131

0.521 ± 0.037

0.825 ± 0.026*

0.235 ± 0.003

1.040 ± 0.025**

0.295 ± 0.002

11.5 ± 0.352*

3.2 ± 0.043**

0.501 ± 0.163**

0.143 ± 0.003**

0.977 ± 0.022**

0.279 ± 0.007

0.982 ± 0.017**

0.281 ± 0.007

17.5 ± 1.370

5.0 ± 0.481

17.0 ± 0.654

4.8 ± 0.146

1.455 ± 0.027**

0.415 ± 0.004**

1.491 ± 0.033**

0.425 ± 0.004**

-

-

-

-

2.205 ± 0.141 **

0.647 ± 0.032**

-

-

0.07 W

1.995 ± 0.016

0.471 ± 0.006

0.981 ± 0.031

0.232 ± 0.008

1.287 ± 0.021

0.304 ± 0.007

13.2 ± 0.258

3.0 ± 0.052

0.682 ± 0.009

0.161 ± 0.002

1.221 ± 0.018

0.288 ± 0.005

1.254 ± 0.012

0.296 ± 0.005*

18.6 ± 0.680

4.4 ± 0.156

19.2 ± 1.024

4.5 ± 0.262

1.606 ± 0.022

0.379 ± 0.006

1.566 ± 0.056

0.370 ± 0.015

-

-

-

-

1.338 ± 0.040

0.314 ± 0.009

-

-

0.2 W

1.881 ± 0.108

0.478 ± 0.028

0.975 ± 0.183

0.247 ± 0.003

1.154 ± 0.037*

0.293 ± 0.007

11.6 ± 0.184

2.9 ± 0.042

0.595 ± 0.013**

0.150 ± 0.002

1.124 ± 0.025

0.286 ± 0.007

1.154 ± 0.031

0.293 ± 0.008

18.4 ± 0.914

4.6 ± 0.232

17.5 ± 1.309

4.4 ± 0.354

1.483 ± 0.059

0.377 ± 0.016

1.589 ± 0.013

0.404 ± 0.005

-

-

-

-

1.210 ± 0.035

0.309 ± 0.008

-

-

Female

Control

1.853 ± 0.023

0.846 ± 0.016

0.647 ± 0.013

0.294 ± 0.003

0.870 ± 0.026

0.397 ± 0.012

6.2 ± 0.150

2.8 ± 0.051

0.441 ± 0.016

0.201 ± 0.006

0.650 ± 0.021

0.296 ± 0.006

0.650 ± 0.021

0.296 ± 0.008

24.8 ± 1.381

11.2 ± 0.443

25.7 ± 1.385

11.7 ± 0.487

-

-

-

-

29.3 ± 2.738

13.4 ± 1.226

31.2 ± 4.007

14.2 ± 1.857

0.828 ± 0.010

0.377 ± 0.005

0.687 ± 0.037

0.313 ± 0.017

0.6 F

1.859 ± 0.011

0.845 ± 0.027

0.595 ± 0.019*

0.269 ± 0.006**

0.810 ± 0.032

0.367 ± 0.015

6.6 ± 0.355

2.9 ± 0.061

0.425 ± 0.010

0.193 ± 0.009

0.648 ± 0.023

0.292 ± 0.004

0.655 ± 0.026

0.296 ± 0.008

25.8 ± 1.563

11.7 ± 0.820

24.8 ± 1.641

11.3 ± 0.863

-

-

-

-

23.3 ± 3.504

10.8 ± 1.740

32.8 ± 3.120

14.9 ± 1.380

0.859 ± 0.064

0.381 ± 0.015

0.660 ± 0.033

0.301 ± 0.020

1.8 F

1.788 ± 0.031

0.954 ± 0.015**

0.455 ± 0.008

0.242 ± 0.002

0.761 ± 0.028

0.405 ± 0.009

7.1 ± 0.195**

3.8 ± 0.082**

0.351 ± 0.012 **

0.187 ± 0.004

0.580 ± 0.015

0.309 ± 0.003

0.586 ± 0.021*

0.312 ± 0.007

19.6 ± 1.981*

10.3 ± 0.924

21.3 ± 1.487*

11.3 ± 0.609

-

-

-

-

25.3 ± 2.853

13.4 ± 1.417

31.5 ± 2.322

16.7 ± 1.165

1.436 ± 0.080**

0.775 ± 0.039**

0.588 ± 0.066

0.312 ± 0.034

0.07 W

1.854 ± 0.019

0.844 ± 0.016

0.611 ± 0.011*

0.277 ± 0.004**

0.801 ± 0.027

0.365 ± 0.014

6.3 ± 0.139

2.8 ± 0.063

0.424 ± 0.007

0.193 ± 0.004

0.669 ± 0.017

0.304 ± 0.006

0.677 ± 0.010

0.308 ± 0.005

22.0 ± 1.619

10.0 ± 0.720

21.0 ± 1.763

9.8 ± 0.790

-

-

-

-

30.0 ± 1.490

13.6 ± 0.780

31.7 ± 2.300

12.2 ± 1.200

0.785 ± 0.051

0.358 ± 0.024

0.729 ± 0.051

0.331 ± 0.022

0.2 W

1.691 ± 0.112

0.810 ± 0.053

0.593 ± 0.015

0.287 ± 0.004

0.817 ± 0.040

0.393 ± 0.021

6.1 ± 0.220

2.9 ± 0.071

0.428 ± 0.014

0.205 ± 0.006

0.651 ± 0.015

0.312 ± 0.006

0.670 ± 0.020

0.321 ± 0.008*

23.2 ± 1.495

11.1 ± 0.677

24.3 ± 1.154

11.6 ± 0.431

-

-

-

-

28.4 ± 1.633

13.6 ± 0.821

28.6 ± 1.900

13.7 ± 0.830

0.800 ± 0.045

0.379 ± 0.026

0.644 ± 0.032

0.308 ± 0.014

Values: mean ± S.E.

*p<0.05

**p<0.01

Table 5: Liver enzyme activities of rats on administration of LAS for 9 months (Yoneyama et. al., 1976)

Sex	Groups	G6 Pase (µmoles Pi/mg protein /hr	G6P-DH (x 10^-3)ΔE/mg protein / min	LDH ΔE/mg protein / min	GPT (x 10^2) KU/mg protein	GOT (x 10^2) KU/mg protein
Male	Control	3.77 ± 0.22	60.3 ± 9.9	11.33 ± 0.53	4.06 ± 0.345	5.80 ± 0.525
	0.6 F	3.82 ± 0.50	40.9 ± 9.1 **	10.28 ± 0.84*	3.94 ± 0.484	5.49 ± 0.810
	1.8 F	2.60 ± 0.015**	36.0 ± 7.7**	9.26 ± 0.99**	3.09 ± 0.225**	4.95 ± 0.510**
	0.07 W	3.57 ± 0.36	68.0 ± 15.1	10.92 ± 0.91	3.89 ± 0.360	5.61 ± 0.410
	0.2 W	3.71 ± 0.35	56.8 ± 14.9	9.28 ± 0.71**	4.10 ± 0.420	6.16 ± 0.545
Female	Control	2.73 ± 0.57	143.6 ± 30.0	70.6 ± 11.6	2.71 ± 0.405	4.91 ± 0.635
	0.6 F	2.67 ± 0.51	100.7 ± 39.9*	63.4 ± 14.2	2.60 ± 0.505	4.30 ± 0.560
	1.8 F	2.17 ± 0.15*	58.5 ± 12.2*	62.2 ± 4.7	2.37 ± 0.225	4.05 ± 0.450 **
	0.07 W	2.61 ± 0.20	138.3 ± 41.0	65.8 ± 8.5	2.57 ± 2.290	5.61 ± 0.410*
	0.2 W	2.76 ± 0.28	120.9 ± 27.6	66.5 ± 11.3	2.89 ± 0.195	6.16 ± 0.545**

Values: mean ± S.D.

*p<0.05

**p<0.01

Table 6: Kidney enzyme activities of rats on administration of LAS for 9 months  (Yoneyama et. al., 1976)

Sex	Groups	ALP (µmoles/mg/hr)	ACP (µmoles/mg/hr)	G6Pase (µmoles/mg/hr)	Na, K-A Tpase (µmoles/mg/hr)	GPT (KU/mL)	GOT (KU/mL)	LDH (ΔE/mg/min)	MDH (ΔE/mg/min)
Male	Control	16.37 ± 1.984	1.80 ± 0.188	3.97 ± 0.590	3.94 ± 0.613	23.99 ± 5.300	192.11 ± 35.741	0.339 ± 0.043	2.308 ± 0.0447
	0.6 F	14.44 ± 2.046	1.72 ± 0.141	3.92 ± 0.618	3.63 ± 0.640	23.16 ± 5.885	199.95 ± 22.869	0.334 ± 0.050	2.325 ± 0.575
	1.8 F	15.74 ± 1.933	1.78 ± 0.179	3.47 ± 0.732	3.32 ± 1.311	21.11 ± 4.690	194.18 ± 13.668	0.358 ± 0.070	2.354 ± 0.554
	0.07 W	14.73 ± 2.387	1.85 ± 0.194	4.26 ± 0.547	3.52 ± 1.170	23.85 ± 4.640	207.39 ± 21.308	0.347 ± 0.098	2.617 ± 0.628
	0.2 W	15.03 ± 1.169	1.66 ± 0.109	4.03 ± 0.428	3.09 ± 0.224*	24.44 ± 3.535	202.94 ± 15.708	0.362 ± 0.058	2.679 ± 0.740
Female	Control	15.94 ± 3.369	2.02 ± 0.182	3.96 ± 0.545	4.29 ± 1.027	28.27 ± 4.407	188.33 ± 21.189	0.350 ± 0.035	2.387 ± 0.601
	0.6 F	17.11 ± 2.919	2.03 ± 0.179	3.63 ± 0.521	4.29 ± 0.656	30.26 ± 5.736	196.48 ± 21.273	0.338 ± 0.044	2.231 ± 0.433
	1.8 F	16.86 ± 2.369	1.85 ± 0.195	3.13 ± 0.426**	3.19 ± 0.613*	32.60 ± 3.737	184.89 ± 11.276	0.291 ± 0.025**	2.181 ± 0.404
	0.07 W	16.24 ± 2.381	1.98 ± 0.218	3.72 ± 0.443	4.14 ± 1.452	27.19 ± 3.319	198.09 ± 29.803	0.322 ± 0.035	2.052 ± 0.306
	0.2 W	15.02 ± 3.244	1.90 ± 0.136	3.45 ± 0.651	4.59 ± 1.154	27.03 ± 4.430	187.36 ± 20.235	0.316 ± 0.052	2.070 ± 0.362

Values: mean ± S.D.

*p<0.05

**p<0.01

Applicant's summary and conclusion

Conclusions:

Administration of LAS to Wistar JCL rats by test diets at dose levels of 0, 0.6 and 1.8% for 9 months (focusing on the liver and kidneys) revealed an LOAEL of 0.6% (300 mg/kg bw/day in diet), based on adverse effects at all dose levels.
Administration of LAS to Wistar JCL rats in drinking water at dose levels of 0, 0.07 and 0.2% for 9 months (focusing on the liver and kidneys) revealed an NOAEL of 0.07% (85 mg/kg bw/day in drinking water), based on adverse effects at all dose levels, based on significant decreases in the activities of glutamate-oxalate transaminase and lactate dehydrogenase in males and significant decrease in renal Na,K-ATPase in males and females at 145 mg/kg bw/day (0.2% drinking water).

Executive summary:

A sub-chronic (9 months) repeated dose toxicity study was conducted with C10-14 LAS, sodium salt in male and female Wistar JCL rats. The test substance was administered at the following dose levels: through diet at 0, 0.6 and 1.8% (equivalent to 0, 300 and 900 mg/kg bw/day) to 8 animals/sex/dose and via drinking water at 0, 0.07 and 0.2% (equivalent to 0, 85 and 145 mg/kg bw/day to 9 animals/sex/dose. Rats at the 0.6 and 1.8% dose group exhibited severe weight loss so LAS administration via diet was stopped after 2 weeks. Clinical observations, water consumption, food consumption and body weights were recorded weekly. Terminal blood collection was done for estimation of haematological and clinical chemistry parameters. Gross findings were observed after euthanizing animals and organs were removed for organ weight measurements. No histopathology was performed. Body weight gain was reduced at the highest dose group only. Significant decreases in transaminase activity and renal Na,K-ATPase were seen at 145 mg/kg bw/day. No significant haematological or organ weight changes were noted. Based on significant decrease in the liver and renal enzyme levels at 145 mg/kg bw/day, the systemic toxicity NOAEL was established at 85 mg/kg bw/day (Yoneyama, 1976).