Registration Dossier
Registration Dossier
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EC number: 284-664-9 | CAS number: 84961-74-0
Endpoint summary
Administrative data
Description of key information
No repeated dose toxicity data are available for the substance LAS IPA.
The endpoint was addressed with information from LAS Na and IPA.
There are three studies with LAS Na: the lowest LOAEL is from the 6-month rat study (Yoneyama et al., 1972) with a LOAEL of 115 mg/kg bw. The highest NOAEL below this LOAEL is from the 9-month rat study (Yoneyama et al., 1976) with NOAEL = 85 mg/kg bw, based on histological changes seen in the kidney. Based on the data from all the studies, an overall NOAEL of 85 mg/kg bw was considered as the most releveant, derived from a 9- month oral study and corresponding to the NOAEL closest to the lowest oral LOAEL of 115 mg/kg bw.
Oral data are not available for IPA; a 90-day toxicity study available is via inhalation. Histological treatment related adverse effect observed was a local effect on the nose, that is expected to be caused by isopropylamine (free amine) and not to the ammonium cation. Other effects seen were decreased body weights and decreased serum glucose in high level female animals, as well as increases in the adrenal weights. The NOAEC of the study for the systemic effects of IPA is set at 100 mg/m3.
Key value for chemical safety assessment
Repeated dose toxicity: via oral route - systemic effects
Link to relevant study records
- Endpoint:
- sub-chronic toxicity: oral
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Study period:
- Unavailable - orginal publication date 1976
- Reliability:
- 3 (not reliable)
- Rationale for reliability incl. deficiencies:
- significant methodological deficiencies
- 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 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, hematological, 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
- Species:
- rat
- Strain:
- Wistar
- Remarks:
- 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 - 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:
- unchanged (no vehicle)
- Analytical verification of doses or concentrations:
- no
- Duration of treatment / exposure:
- Nine months
- Frequency of treatment:
- Continuous in diet or drinking water (ad libitum)
- Dose / conc.:
- 300 mg/kg bw/day (nominal)
- Remarks:
- In diet (corresponding to 0.6% dose level)
- Dose / conc.:
- 900 mg/kg bw/day (nominal)
- Remarks:
- In diet (corresponding to 1.8% dose level)
- Dose / conc.:
- 0.07 other: %
- Remarks:
- In drinking water
- Dose / conc.:
- 0.2 other: %
- Remarks:
- In drinking water; 0.6 and 1.8% dose group (equivalent to 857.14 and 2571.43 mg/kg bw/day) were 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): 8 animals/sex/dose
Drinking water study: 9 animals/sex/dose - 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: 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), Hemoglobin (Hgb), Hematocrit (Hct), mean corpuscular volume (MCV) and mean corpuscular hemoglobin 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 - Clinical signs:
- effects observed, treatment-related
- Description (incidence and severity):
- Both female and male rats (drinking water study) 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 (Table 1).
- 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 (Table 1).
- 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 and 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):
- There were significant alteration in cholesterol [decrease; all doses, except female rats of 0.07% dose group (drinking water)], GPT [0.6% dose group (diet) females)], GOT [1.8% dose group (diet) males], albumin [1.8% (diet) males] , ALP levels [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 (Table 2).
There was also significant decrease in absolute and relative heart weight in females of 0.6% (diet) and 0.07% (drinking water) dose groups (Table 2). - 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):
- 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. - Details on results:
- CLINICAL SIGNS: 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.
BODY WEIGHT AND WEIGHT CHANGES: There was significant decrease in body weight gain in male and females of 1.8% dose group fed with diet and in males of 0.2% dose group consuming LAS contained water.
WATER CONSUMPTION AND COMPOUND INTAKE: Increased water consumption was observed in males of 1.8% dose group fed with diet and 0.2% dose group consuming LAS contained water.
HAEMATOLOGICAL FINDINGS: 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: 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.
ORGAN WEIGHT: 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.
LIVER ENZYME TESTS: G6Pase activity was reduced in 1.8% dose group (diet) males and females, G6P-DH activity was reduced in 0.6 and 1.8% dose group (diet) males and females, where the percentage reduction was greater in 1.8% dose group (diet) animals. LDH activity was clearly reduced in 0.6, 1.8% dose group (diet), and 0.2% dose group (drinking water) males, but reduced in only 1.8% dose group (diet) females. GOT and GPT activities were clearly reduced in males, while in females GPT activity was reduced only in 1.8% dose group (diet) animals, and GOT activity was increased in 0.07 and 0.2% dose group (drinking water) animals but reduced in 1.8% dose group (diet) animals.
RENAL ENZYME TESTS: In males, Na, K-ATPase activity was significantly reduced in 0.2% dose group (drinking water) animals, and also reduced in other male treatment groups. A significant difference was also observed in G6Pase activity, where the reduction observed was associated with an increase in amount consumed. In females, G6Pase, Na, K-ATPase, and LDH activity were significantly reduced in 1.8% dose group (diet) animals, and G6Pase and LDH activity were also reduced in other treatment groups. - Dose descriptor:
- NOAEL
- Effect level:
- 85 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Remarks:
- Drinking water
- 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 lactat e 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).
- Dose descriptor:
- LOAEL
- Effect level:
- 300 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Remarks:
- Dietary 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
- 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
- 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
- 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 a 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 a 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:
The 9 months sub-chronic oral toxicity study of LAS was performed in Wistar JCL rats, focusing on the liver and kidneys.
Four week old male and female Wistar JCL rats (obtained from Shizuoka Agricultural Cooperative Association for Laboratory Animals) with body weight range100 - 124 g (males), 82 - 100 g (females) were used in the study. Five animals were housed in each cage and maintained under controlled environmental conditions (temperature: Average of 25 ± 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, 0.6 and 1.8% (equivalent to 0, 300 and 900 mg/kg bw/day); 8 animals/sex/dose
Dissolved in drinking water: 0, 0.07 and 0.2 % (equivalent to 0,85 and 145mg/kg bw/day); 9 animals/sex/dose
Rats in 0.6 and 1.8% dose group of 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 observed 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 appendixwere recorded. Liver and kidney enzymes were also analysed. No histopathology was performed.
No mortality was observed throughout the study. Both female and male rats consuming LAS-containing water exhibited a slight redness at the tips of their facial fur, and coarse fur over their bodies. There was significant decrease in body weight gain in male and females of 1.8% dose group fed with diet and in males of 0.2% dose group consuming LAS contained water. Increased water consumption was observed in males of 1.8% dose group fed with diet and 0.2% dose group consuming LAS contained water.
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. A marked reduction in cholesterol was observed in male and female rats of all dose groups [except female rats of 0.07% dose group (drinking water)] compared to controls. This indicate hepatocyte damage. 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) male 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.
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.
Liver enzymes were markedly reduced in 1.8% fed rats, due to impaired liver function, indicates reduced enzyme synthesis and direct enzyme inhibition by LAS or its metabolites. Renal G6Pase and Na, KATPase activity decreased, indicating kidney impairment.
Administration of LAS to Wistar JCL rats bytest 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 onsignificant 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).
- Endpoint:
- short-term repeated dose toxicity: oral
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Study period:
- Unavailable - orginal publication date 1978
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- study well documented, meets generally accepted scientific principles, acceptable for assessment
- Remarks:
- This study is assigned a reliability score of 2 because the original report was not available for review. However, the study was evaluated by IPCS prior to inclusion in their criteria document.
- Qualifier:
- no guideline followed
- Principles of method if other than guideline:
- The purpose of this study is to determine the sub-acute toxicity of C10-13 sodium linear alkyl benzene sulfonate (LAS-Na) in CRJ-SD rats. LAS-Na was orally administered at 0 (distilled water), 125, 250 and 500 mg/kg bw to male and females rats for 29 and 30 days, respectively. General symptoms of toxicity and the body weights were recorded daily while the food consumption was measured twice a week. Urinalysis was also performed during the study. At the end of study, blood was collected for estimation of haematological and clinical chemistry parameters. After collection of blood samples, the animals were humanely euthanized, gross necropsy was performed and organs weights were determined.
- GLP compliance:
- no
- Remarks:
- Pre-dates GLP
- 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 Laboratories Japan, Inc.
- Age at study initiation: 5 weeks
- Weight at study initiation: Not reported
- Fasting period before study: Not reported
- Housing: 3 male and 4 female animals per cage.
- Diet: Solid feed (CE-2, CLEA Japan, Inc.); ad libitum
- Water: Tap water; ad libitum
- Acclimation period: After 1 week pre-housing, the animals were used in the experiment.
ENVIRONMENTAL CONDITIONS
- Temperature: 23 ± 2°C
- Humidity: 55 ± 5%
- Air changes: Not reported
- Photoperiod: Not reported - Route of administration:
- oral: gavage
- Details on route of administration:
- The test substance was administered orally through a metallic gastric sonde.
- Vehicle:
- not specified
- Details on oral exposure:
- PREPARATION OF DOSING SOLUTIONS: The test substance was suspended in distilled water prior to dosing.
- DOSE VOLUME: 5 mL/kg bw
- Concentration in vehicle: 0, 250, 500, and 1000 mg/mL respectively for administration of doses 0, 125, 250 and 500 mg/kg bw. - Analytical verification of doses or concentrations:
- no
- Duration of treatment / exposure:
- 29 days for males and 30 days for females
- Frequency of treatment:
- Once daily
- Dose / conc.:
- 125 mg/kg bw/day (actual dose received)
- Remarks:
- In distilled water (corresponding to 250 mg/mL)
- Dose / conc.:
- 250 mg/kg bw/day (actual dose received)
- Remarks:
- In distilled water (corresponding to 500 mg/mL)
- Dose / conc.:
- 500 mg/kg bw/day (actual dose received)
- Remarks:
- In distilled water (corresponding to 1000 mg/mL)
- No. of animals per sex per dose:
- Test group: 12 animals/sex/dose group
Control group: 15 male/group and 16 female/group - Control animals:
- yes, concurrent vehicle
- Details on study design:
- Dose selection rationale: The dose levels for this study were selected based on the results of acute toxicity study conducted with LAS-Na in rats. 1/3 of the LD50 levels of LAS-Na, i.e. 500 mg/kg bw, was selected as the highest dose. The other two doses were set at common ratios of 2, whereby the medium and low doses were 250 (1/6 LD50) and 125 (1/12 LD50) mg/kg bw, respectively.
- Positive control:
- No.
- Observations and examinations performed and frequency:
- CLINICAL OBSERVATIONS: Yes
- Time schedule: Daily
BODY WEIGHT: Yes
- Time schedule for examinations: Daily
FOOD CONSUMPTION AND COMPOUND INTAKE:
- Food consumption: Yes
- Time schedule for examinations: Twice/week
FOOD EFFICIENCY: Yes
WATER CONSUMPTION AND COMPOUND INTAKE: No
OPHTHALMOSCOPIC EXAMINATION: No
HAEMATOLOGY: Yes
- Time schedule for collection of blood: After 1 month, blood was collected was collected from the carotid artery using a cannula.
- Anesthetic used for blood collection: Yes, light ether anesthesia was used.
- Animals fasted: Not reported
- How many animals: All surviving animals
- Parameters: Hemoglobin, hematocrit, red and white blood cell count were measured using an autoanalyzer SMA-4A.
CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: After 1 month, blood was collected from each animal and centrifuged (at 3000 rpm for 15 minutes) to obtain the serum samples for analysis.
- Animals fasted: No
- How many animals: All surviving animals
- Parameters: s-GOT, s-GPT, alkaline phosphatase, total protein, albumin, glucose, cholesterol, bilirubin, creatinine, urea nitrogen, inorganic phosphates and ions (Na+, K+, Ca2+, Mg2+ and Cl-)
URINALYSIS: Yes
- Time schedule: Not reported
- Metabolism cages used for collection of urine: No
- Animals fasted: No
- How many animals: All surviving animals
- Parameters: pH, proteins, sugar, ketone bodies and occult blood
NEUROBEHAVIOURAL EXAMINATION: No
IMMUNOLOGY: No - Sacrifice and pathology:
- GROSS PATHOLOGY: Yes
ORGAN WEIGHTS: The wet weight of liver, kidneys, spleen, adrenal glands, lungs, heart, thymus, reproductive organ, pituitary and the brain were measured. Based on this, the relative weights were calculated.
HISTOPATHOLOGY: No - Clinical signs:
- effects observed, treatment-related
- Description (incidence and severity):
- At 500 mg/kg bw, animals exhibited the action of wetting the area around the mouth and nose, and rubbing it against the cage. Many animals exhibited diarrhea from the start of dosing as well as a suppressed state, several hours after dosing. Soft stool was observed in some animals dosed at 125 and 250 mg/kg bw, however, the animals showed recovery at low and mid doses.
- Mortality:
- no mortality observed
- Body weight and weight changes:
- effects observed, treatment-related
- Description (incidence and severity):
- Male animals of all dose groups and female animals of high dose group (i.e. 500 mg/kg bw) exhibit body weight suppression.
- Food consumption and compound intake (if feeding study):
- effects observed, treatment-related
- Description (incidence and severity):
- Animals showed a dose dependent decline in the feed consumption.
- Food efficiency:
- effects observed, treatment-related
- Description (incidence and severity):
- Decreased feed efficacy was observed for male animals at 500 mg/kg bw.
- Water consumption and compound intake (if drinking water study):
- not examined
- Ophthalmological findings:
- not examined
- Haematological findings:
- no effects observed
- Clinical biochemistry findings:
- effects observed, treatment-related
- Description (incidence and severity):
- Effects at 125 mg/kg/bw: Decreased total proteins (only females), albumin (only females), Ca2+ levels (only males), and increased cholesterol (only males).
Effects at 250 mg/kg bw: Decreased s-GPT (both males and females), total proteins (only females), albumin (only females), Na+ (only males), K+ (only males), Ca2+ (both males and females), and Mg2+ (only males).
Effects at 500 mg/kg bw: Decreased s-GOT (only females), total proteins (only females), albumin (only females), glucose (only females), Ca2+ (both males and females), and increased alkalinephosphatase (only males) and urea nitrogen (only females). - 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):
- The relative liver weight of female animals was significantly increased at 500 mg/kg bw (p<0.01). Apart from this, increase in relative weight of adrenal glands (p<0.05 for right side; p<0.01 for left side), testes (p<0.05) and brain (p<0.05) was also observed in male animals at 500 mg/kg bw. Decrease in the relative weight of liver (only females; at 125 mg/kg bw, p<0.05), right side kidney (only females; at 250 and 500 mg/kg bw, p<0.05), spleen (only males; at 500 mg/kg bw, p<0.05), heart (both males and females at 500 mg/kg bw, p<0.01; males at 250 mg/kg bw, p<0.05; females at 125 mg/kg bw, p<0.05) and thymus (only females; at 500 mg/kg bw, p<0.01) was also observed. Although, some significant effects were observed at the low and mid dose yet they were not dose-dependent.
- Gross pathological findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- At 500 mg/kg bw, the animals showed hypertrophy in the stomach wall of the proventriculus, as if the process of inflammation and recovery had repeated over and over. Therefore, formation of new capillaries was observed on the peritoneal side. There were no additional changes relative to the control group although, events of hydronephrosis, bleeding plaques on the thymus and pneumonia were observed including the control group.
- Neuropathological findings:
- not examined
- Histopathological findings: non-neoplastic:
- not examined
- Histopathological findings: neoplastic:
- not examined
- Other effects:
- not examined
- Details on results:
- CLINICAL SIGNS: At 500 mg/kg bw, animals exhibited the action of wetting the area around the mouth and nose, and rubbing it against the cage. Many animals exhibited diarrhea from the start of dosing as well as a suppressed state, several hours after dosing. Soft stool was observed in some animals dosed at 125 and 250 mg/kg bw, however, the animals showed recovery at low and mid doses.
MORTALITY: No mortality was observed at any dose level.
BODY WEIGHT AND WEIGHT CHANGES: Body weight suppression was observed in animals (both males and females) belonging to the high dose group (i.e. 500 mg/kg bw).
FOOD CONSUMPTION AND COMPOUND INTAKE: Animals showed a dose dependent decline in the feed consumption. Decreased feed efficacy was observed for male animals at 500 mg/kg bw.
CLINICAL BIOCHEMISTRY: Few significant effects were observed in the biochemical parameters mainly at the mid and high dose level.
URINALYSIS: Results of urinalysis (pH, proteins, sugar, ketone bodies and occult blood) in the test animals were comparable to the control group.
ORGAN WEIGHT: The relative liver weight of female animals was significantly increased at 500 mg/kg bw (p<0.01). Apart from this, increase in relative weight of adrenal glands (p<0.05 for right side; p<0.01 for left side), testes (p<0.05) and brain (p<0.05) was also observed in male animals at 500 mg/kg bw. Decrease in the relative weight of liver (only females; at 125 mg/kg bw, p<0.05), right side kidney (only females; at 250 and 500 mg/kg bw, p<0.05), spleen (only males; at 500 mg/kg bw, p<0.05), heart (both males and females at 500 mg/kg bw, p<0.01; males at 250 mg/kg bw, p<0.05; females at 125 mg/kg bw, p<0.05) and thymus (only females; at 500 mg/kg bw, p<0.01) was also observed. Although, some significant effects were observed at the low and mid dose yet they were not dose-dependent.
GROSS PATHOLOGY: At 500 mg/kg bw, the animals showed hypertrophy in the stomach wall of the proventriculus, as if the process of inflammation and recovery had repeated over and over. Therefore, formation of new capillaries was observed on the peritoneal side. There were no additional changes relative to the control group although, events of hydronephrosis, bleeding plaques on the thymus and pneumonia were observed including the control group. - Key result
- Dose descriptor:
- NOAEL
- Effect level:
- 125 other: mg/kg bw d
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- body weight and weight gain
- clinical biochemistry
- organ weights and organ / body weight ratios
- Remarks on result:
- other: Based on reduced body weight and changes in organ weight (without histopathological correlation), clinical chemistry. The LOAEL was 250 mg/kg bw.
- Dose descriptor:
- LOAEL
- Effect level:
- 250 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- body weight and weight gain
- clinical biochemistry
- organ weights and organ / body weight ratios
- Remarks on result:
- other: Based on reduced body weight and changes in organ weight (without histopathological correlation), clinical chemistry.
- Critical effects observed:
- not specified
- Conclusions:
- Oral administration of C10-13 sodium linear alkylbenzene sulphonate (LAS-Na) to CRJ-SD rats at 0, 125, 250 and 500 mg/kg bw/day for 1 month resulted in a NOAEL of 125 mg/kg bw/day, based on reduced body weight, changes in organ weight (without histopathological correlation) and clinical chemistry at the mid and high doses. The LOAEL was 250 mg/kg bw/day.
- Executive summary:
One month sub-acute oral toxicity study of sodium linear alkylbenzene sulphonate (LAS-Na) was performed in CRJ-SD rats. Five weeks old male and female CRJ-SD rats (obtained from Charles River Laboratories Japan, Inc.) were used in the study. Three males and 4 females were housed in the cages maintained under controlled environmental conditions (temperature: 23 ± 2°C and humidity: 55 ± 5%). Solid feed (CE-2, CLEA Japan, Inc.) and tap water were provided ad libitum. The substance was orally administered (via metallic gastric sonde) at 0 (distilled water), 125, 250 and 500 mg/kg bw/day (corresponding to 250, 500 and 1000 mg/mL) to male and females rats for 29 and 30 days, respectively. General symptoms of toxicity and the body weights were recorded daily while the food consumption was measured twice a week. Urinalysis was also performed during the study. At the end of study, blood was collected for estimation of haematological and clinical chemistry parameters. After collection of blood samples, the animals were humanely euthanized, gross necropsy was performed and the organs weights determined.
No mortality was observed at any dose level. Significant body weight loss was observed at 500 mg/kg bw/day. The relative liver weight of female animals was significantly increased at 500 mg/kg bw/day (p<0.01). Apart from this, increase in relative weight of adrenal glands (p<0.05 for right side; p<0.01 for left side), testes (p<0.05) and brain (p<0.05) was also observed in male animals at 500 mg/kg bw/day. Decrease in the relative weight of liver (only females; at 125 mg/kg bw/day, p<0.05), right side kidney (only females; at 250 and 500 mg/kg bw/day, p<0.05), spleen (only males; at 500 mg/kg bw/day, p<0.05), heart (both males and females at 500 mg/kg bw/day, p<0.01; males at 250 mg/kg bw/day, p<0.05; females at 125 mg/kg bw/day, p<0.05) and thymus (only females; at 500 mg/kg bw/day, p<0.01) was also observed. Although, some significant effects were observed at the low and mid dose yet they were not dose-dependent. Few significant effects were observed in the biochemical parameters mainly at the mid and high dose level.
Based on the results, oral administration of C10-13 sodium linear alkylbenzene sulphonate (LAS-Na) to CRJ-SD rats at 0, 125, 250 and 500 mg/kg bw/day for 1 month resulted in a NOAEL of 125 mg/kg bw/day, based on reduced body weight, changes in organ weight (histopathology was not conducted) and clinical chemistry at the mid and high doses. The LOAEL was 250 mg/kg bw/day.
- Endpoint:
- sub-chronic toxicity: oral
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 1972-05-30 to 1972-11-27
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Acceptable study, followed scientific principles/standards, pre-dates GLP
- Remarks:
- This study is assigned a reliability score of 2 because the original report was not available for review. However, the study was evaluated by IPCS prior to inclusion in their criteria document.
- Qualifier:
- no guideline followed
- Principles of method if other than guideline:
- The purpose of this study is to determine the sub chronic toxicity of C10 - C14 linear alkylbenzenesulfonic 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 haematological 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 was performed.
- GLP compliance:
- no
- Remarks:
- Pre-dates GLP
- Limit test:
- no
- Species:
- rat
- Strain:
- Wistar
- Remarks:
- SLC
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- 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 - 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:
- unchanged (no vehicle)
- Analytical verification of doses or concentrations:
- no
- Duration of treatment / exposure:
- 26 weeks
- Frequency of treatment:
- Continuous in diet (ad libitum)
- Dose / conc.:
- 40 mg/kg bw/day (nominal)
- Remarks:
- In diet (corresponding to 0.07% dose level)
- Dose / conc.:
- 115 mg/kg bw/day (nominal)
- Remarks:
- In diet (corresponding to 0.2% dose level)
- Dose / conc.:
- 340 mg/kg bw/day (nominal)
- Remarks:
- In diet (corresponding to 0.6% dose level)
- 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.
- 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
HAEMATOLOGY: 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 e
uthanasia (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 (all tissues weighed and also stomach, large and small intestines, pancreas, epidiymis, skin and bones) were examined and subjected to microscopy through H.E., PAS and silver staining. - 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 (Table 1).
- 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 (Table 1).
- 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 (Table 1).
- 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.
- 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):
- Organ weight findings are presented in Table 2.
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:
- no effects observed
- Description (incidence and severity):
- No significant changes were observed in the animals.
- 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 animals did not recover fully by the end of the study. The animals moved slowly, and there 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.
HAEMATOLOGY:
- 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 and decrease in total protein.
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 swelling 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 thereon, 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% treated groups)
- Glomeruli contributed to overall swelling through dilation of vessel lumen and interstitial swelling (significant in in male/female rats of 0.2 and 0.07% dose groups. They were not very significant in the 1.8 and 0.6%)
- The glomerular interstitum had thawn mildly but chronically (significant for male/female rats in 1.8 and 0.6% groups)
- Cloudy swelling/vacuolar degeneration of proximal renal tubular epithelial cells were seen (slightly significant in LAS treated groups).
- Small, yellowish brown blobs the size of nuclei were 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
- The renal tubule had collapsed (significantly in male/females of 1.8% dose group)
- Swelling of surrounding connective tissue was observed in the interstitium (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. - Key result
- Dose descriptor:
- NOAEL
- Effect level:
- 40 mg/kg bw/day (nominal)
- Based on:
- test mat.
- 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 clin ical 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).
- 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
- 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 a 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:
The 26 weeks sub-chronic oral toxicity study of linear alkylbenzene sulfonic acid sodium salt (LAS) was performed in Wistar SLC rats. Approximately 4 weeks old male and female Wistar SLC rats (obtained from Shizuoka Agricultural Cooperative Association for Laboratory Animals) with body weight range 80 - 90 g (males), 65 - 80 g (females) were used in the study. Five animals were housed in each stainless steelcage and maintained under controlled environmental conditions (temperature: Average of 25 ± 1°C, humidity:55 - 65%, and 12 hours light /12 hours dark). FII food (from Funabashi Farm) and tap water were provided ad libitum. The animals were administered daily with the LAS in diet 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. 10 animals/sex/dose group was taken in the study.
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 findings were obtained after euthanizing animals and organs were removed for organ weight measurements.Organ weights for brain, pituitary, thyroid, thymus, heart, lungs, liver, kidneys, spleen, adrenal gland, prostate, testes, uterus, ovaries and appendixwere recorded. Liver and kidney enzymes were also analysed. Collected tissues were then examined and subjected to microscopy through H.E., PAS and silver staining. One male animal from the 1.8% dose group died. In the 1.8% dose groups, increase in relative weight of organs associated with diarrhea and significant suppression of weight gain has been remarked. There was also anemia and an increasing trend of white blood cell count, in addition to increased serum ALP activity, decrease in serum total protein and a very high level of tissue damage in the kidneys. In the 0.6% dose groups, there were mild suppression of weight gain, increased weight of appendix as well as fluctuations in serum ALP / total protein and tissue damage to the kidneys. In the 0.2% dose groups, there were increase in weight of appendix and histological abnormalities in a part of the kidneys. In the 0.2% dose groups, there were increase in weight of appendix and histological abnormalities in a part of the kidneys. 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. Administration of linear alkylbenzene sulfonic acid sodium salt (LAS) to Wistar SLC ratsat 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 identified a NOAEL of 0.07% (40 mg/kg bw/day), based on tissue damage in caecum, kidney and liver, hematological as well as clinical chemistry parameters changes at higher dose levels (115 mg/kg bw/day and above).
- Endpoint:
- short-term repeated dose toxicity: oral
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- weight of evidence
- Justification for type of information:
- Read-across approach - see read-across justification in section 13.
- Reason / purpose for cross-reference:
- read-across source
- Principles of method if other than guideline:
- The purpose of this study is to determine the sub-acute toxicity of C10-13 sodium linear alkyl benzene sulfonate (LAS-Na) in CRJ-SD rats. LAS-Na was orally administered at 0 (distilled water), 125, 250 and 500 mg/kg bw to male and females rats for 29 and 30 days, respectively. General symptoms of toxicity and the body weights were recorded daily while the food consumption was measured twice a week. Urinalysis was also performed during the study. At the end of study, blood was collected for estimation of haematological and clinical chemistry parameters. After collection of blood samples, the animals were humanely euthanized, gross necropsy was performed and organs weights were determined.
- GLP compliance:
- no
- Remarks:
- Pre-dates GLP
- 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 Laboratories Japan, Inc.
- Age at study initiation: 5 weeks
- Weight at study initiation: Not reported
- Fasting period before study: Not reported
- Housing: 3 male and 4 female animals per cage.
- Diet: Solid feed (CE-2, CLEA Japan, Inc.); ad libitum
- Water: Tap water; ad libitum
- Acclimation period: After 1 week pre-housing, the animals were used in the experiment.
ENVIRONMENTAL CONDITIONS
- Temperature: 23 ± 2°C
- Humidity: 55 ± 5%
- Air changes: Not reported
- Photoperiod: Not reported - Route of administration:
- oral: gavage
- Details on route of administration:
- The test substance was administered orally through a metallic gastric sonde.
- Vehicle:
- not specified
- Details on oral exposure:
- PREPARATION OF DOSING SOLUTIONS: The test substance was suspended in distilled water prior to dosing.
- DOSE VOLUME: 5 mL/kg bw
- Concentration in vehicle: 0, 250, 500, and 1000 mg/mL respectively for administration of doses 0, 125, 250 and 500 mg/kg bw. - Analytical verification of doses or concentrations:
- no
- Duration of treatment / exposure:
- 29 days for males and 30 days for females
- Frequency of treatment:
- Once daily
- Dose / conc.:
- 125 mg/kg bw/day (actual dose received)
- Remarks:
- In distilled water (corresponding to 250 mg/mL)
- Dose / conc.:
- 250 mg/kg bw/day (actual dose received)
- Remarks:
- In distilled water (corresponding to 500 mg/mL)
- Dose / conc.:
- 500 mg/kg bw/day (actual dose received)
- Remarks:
- In distilled water (corresponding to 1000 mg/mL)
- No. of animals per sex per dose:
- Test group: 12 animals/sex/dose group
Control group: 15 male/group and 16 female/group - Control animals:
- yes, concurrent vehicle
- Details on study design:
- Dose selection rationale: The dose levels for this study were selected based on the results of acute toxicity study conducted with LAS-Na in rats. 1/3 of the LD50 levels of LAS-Na, i.e. 500 mg/kg bw, was selected as the highest dose. The other two doses were set at common ratios of 2, whereby the medium and low doses were 250 (1/6 LD50) and 125 (1/12 LD50) mg/kg bw, respectively.
- Positive control:
- No.
- Observations and examinations performed and frequency:
- CLINICAL OBSERVATIONS: Yes
- Time schedule: Daily
BODY WEIGHT: Yes
- Time schedule for examinations: Daily
FOOD CONSUMPTION AND COMPOUND INTAKE:
- Food consumption: Yes
- Time schedule for examinations: Twice/week
FOOD EFFICIENCY: Yes
WATER CONSUMPTION AND COMPOUND INTAKE: No
OPHTHALMOSCOPIC EXAMINATION: No
HAEMATOLOGY: Yes
- Time schedule for collection of blood: After 1 month, blood was collected was collected from the carotid artery using a cannula.
- Anesthetic used for blood collection: Yes, light ether anesthesia was used.
- Animals fasted: Not reported
- How many animals: All surviving animals
- Parameters: Hemoglobin, hematocrit, red and white blood cell count were measured using an autoanalyzer SMA-4A.
CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: After 1 month, blood was collected from each animal and centrifuged (at 3000 rpm for 15 minutes) to obtain the serum samples for analysis.
- Animals fasted: No
- How many animals: All surviving animals
- Parameters: s-GOT, s-GPT, alkaline phosphatase, total protein, albumin, glucose, cholesterol, bilirubin, creatinine, urea nitrogen, inorganic phosphates and ions (Na+, K+, Ca2+, Mg2+ and Cl-)
URINALYSIS: Yes
- Time schedule: Not reported
- Metabolism cages used for collection of urine: No
- Animals fasted: No
- How many animals: All surviving animals
- Parameters: pH, proteins, sugar, ketone bodies and occult blood
NEUROBEHAVIOURAL EXAMINATION: No
IMMUNOLOGY: No - Sacrifice and pathology:
- GROSS PATHOLOGY: Yes
ORGAN WEIGHTS: The wet weight of liver, kidneys, spleen, adrenal glands, lungs, heart, thymus, reproductive organ, pituitary and the brain were measured. Based on this, the relative weights were calculated.
HISTOPATHOLOGY: No - Clinical signs:
- effects observed, treatment-related
- Description (incidence and severity):
- At 500 mg/kg bw, animals exhibited the action of wetting the area around the mouth and nose, and rubbing it against the cage. Many animals exhibited diarrhea from the start of dosing as well as a suppressed state, several hours after dosing. Soft stool was observed in some animals dosed at 125 and 250 mg/kg bw, however, the animals showed recovery at low and mid doses.
- Mortality:
- no mortality observed
- Body weight and weight changes:
- effects observed, treatment-related
- Description (incidence and severity):
- Male animals of all dose groups and female animals of high dose group (i.e. 500 mg/kg bw) exhibit body weight suppression.
- Food consumption and compound intake (if feeding study):
- effects observed, treatment-related
- Description (incidence and severity):
- Animals showed a dose dependent decline in the feed consumption.
- Food efficiency:
- effects observed, treatment-related
- Description (incidence and severity):
- Decreased feed efficacy was observed for male animals at 500 mg/kg bw.
- Water consumption and compound intake (if drinking water study):
- not examined
- Ophthalmological findings:
- not examined
- Haematological findings:
- no effects observed
- Clinical biochemistry findings:
- effects observed, treatment-related
- Description (incidence and severity):
- Effects at 125 mg/kg/bw: Decreased total proteins (only females), albumin (only females), Ca2+ levels (only males), and increased cholesterol (only males).
Effects at 250 mg/kg bw: Decreased s-GPT (both males and females), total proteins (only females), albumin (only females), Na+ (only males), K+ (only males), Ca2+ (both males and females), and Mg2+ (only males).
Effects at 500 mg/kg bw: Decreased s-GOT (only females), total proteins (only females), albumin (only females), glucose (only females), Ca2+ (both males and females), and increased alkalinephosphatase (only males) and urea nitrogen (only females). - 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):
- The relative liver weight of female animals was significantly increased at 500 mg/kg bw (p<0.01). Apart from this, increase in relative weight of adrenal glands (p<0.05 for right side; p<0.01 for left side), testes (p<0.05) and brain (p<0.05) was also observed in male animals at 500 mg/kg bw. Decrease in the relative weight of liver (only females; at 125 mg/kg bw, p<0.05), right side kidney (only females; at 250 and 500 mg/kg bw, p<0.05), spleen (only males; at 500 mg/kg bw, p<0.05), heart (both males and females at 500 mg/kg bw, p<0.01; males at 250 mg/kg bw, p<0.05; females at 125 mg/kg bw, p<0.05) and thymus (only females; at 500 mg/kg bw, p<0.01) was also observed. Although, some significant effects were observed at the low and mid dose yet they were not dose-dependent.
- Gross pathological findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- At 500 mg/kg bw, the animals showed hypertrophy in the stomach wall of the proventriculus, as if the process of inflammation and recovery had repeated over and over. Therefore, formation of new capillaries was observed on the peritoneal side. There were no additional changes relative to the control group although, events of hydronephrosis, bleeding plaques on the thymus and pneumonia were observed including the control group.
- Neuropathological findings:
- not examined
- Histopathological findings: non-neoplastic:
- not examined
- Histopathological findings: neoplastic:
- not examined
- Other effects:
- not examined
- Details on results:
- CLINICAL SIGNS: At 500 mg/kg bw, animals exhibited the action of wetting the area around the mouth and nose, and rubbing it against the cage. Many animals exhibited diarrhea from the start of dosing as well as a suppressed state, several hours after dosing. Soft stool was observed in some animals dosed at 125 and 250 mg/kg bw, however, the animals showed recovery at low and mid doses.
MORTALITY: No mortality was observed at any dose level.
BODY WEIGHT AND WEIGHT CHANGES: Body weight suppression was observed in animals (both males and females) belonging to the high dose group (i.e. 500 mg/kg bw).
FOOD CONSUMPTION AND COMPOUND INTAKE: Animals showed a dose dependent decline in the feed consumption. Decreased feed efficacy was observed for male animals at 500 mg/kg bw.
CLINICAL BIOCHEMISTRY: Few significant effects were observed in the biochemical parameters mainly at the mid and high dose level.
URINALYSIS: Results of urinalysis (pH, proteins, sugar, ketone bodies and occult blood) in the test animals were comparable to the control group.
ORGAN WEIGHT: The relative liver weight of female animals was significantly increased at 500 mg/kg bw (p<0.01). Apart from this, increase in relative weight of adrenal glands (p<0.05 for right side; p<0.01 for left side), testes (p<0.05) and brain (p<0.05) was also observed in male animals at 500 mg/kg bw. Decrease in the relative weight of liver (only females; at 125 mg/kg bw, p<0.05), right side kidney (only females; at 250 and 500 mg/kg bw, p<0.05), spleen (only males; at 500 mg/kg bw, p<0.05), heart (both males and females at 500 mg/kg bw, p<0.01; males at 250 mg/kg bw, p<0.05; females at 125 mg/kg bw, p<0.05) and thymus (only females; at 500 mg/kg bw, p<0.01) was also observed. Although, some significant effects were observed at the low and mid dose yet they were not dose-dependent.
GROSS PATHOLOGY: At 500 mg/kg bw, the animals showed hypertrophy in the stomach wall of the proventriculus, as if the process of inflammation and recovery had repeated over and over. Therefore, formation of new capillaries was observed on the peritoneal side. There were no additional changes relative to the control group although, events of hydronephrosis, bleeding plaques on the thymus and pneumonia were observed including the control group. - Key result
- Dose descriptor:
- NOAEL
- Effect level:
- 125 other: mg/kg bw d
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- body weight and weight gain
- clinical biochemistry
- organ weights and organ / body weight ratios
- Remarks on result:
- other: Based on reduced body weight and changes in organ weight (without histopathological correlation), clinical chemistry. The LOAEL was 250 mg/kg bw.
- Critical effects observed:
- not specified
- Conclusions:
- Oral administration of C10-13 sodium linear alkylbenzene sulphonate (LAS-Na) to CRJ-SD rats at 0, 125, 250 and 500 mg/kg bw for 1 month resulted in a NOAEL of 125 mg/kg bw, based on reduced body weight, changes in organ weight (without histopathological correlation) and clinical chemistry at the mid and high doses. The LOAEL was 250 mg/kg bw.
- Executive summary:
One month sub-acute oral toxicity study of sodium linear alkylbenzene sulphonate (LAS-Na) was performed in CRJ-SD rats. Five weeks old male and female CRJ-SD rats (obtained from Charles River Laboratories Japan, Inc.) were used in the study. Three males and 4 females were housed in the cages maintained under controlled environmental conditions (temperature: 23 ± 2°C and humidity: 55 ± 5%). Solid feed (CE-2, CLEA Japan, Inc.) and tap water were provided ad libitum. The substance was orally administered (via metallic gastric sonde) at 0 (distilled water), 125, 250 and 500 mg/kg bw (corresponding to 250, 500 and 1000 mg/mL) to male and females rats for 29 and 30 days, respectively. General symptoms of toxicity and the body weights were recorded daily while the food consumptionwas measured twice a week. Urinalysis was also performed during the study. At the end of study, blood was collected for estimation of haematological and clinical chemistry parameters. After collection of blood samples, the animals were humanely euthanized, gross necropsy was performed and the organs weights determined. No mortality was observed at any dose level. Significant body weight loss was observed at 500 mg/kg bw. The relative liver weight of female animals was significantly increased at 500 mg/kg bw (p<0.01). Apart from this, increase in relative weight of adrenal glands (p<0.05 for right side; p<0.01 for left side), testes (p<0.05) and brain (p<0.05) was also observed in male animals at 500 mg/kg bw. Decrease in the relative weight of liver (only females; at 125 mg/kg bw, p<0.05), right side kidney (only females; at 250 and 500 mg/kg bw, p<0.05), spleen (only males; at 500 mg/kg bw, p<0.05), heart (both males and females at 500 mg/kg bw, p<0.01; males at 250 mg/kg bw, p<0.05; females at 125 mg/kg bw, p<0.05) and thymus (only females; at 500 mg/kg bw, p<0.01) was also observed. Although, some significant effects were observed at the low and mid dose yet they were not dose-dependent. Few significant effects were observed in the biochemical parameters mainly at the mid and high dose level. Based on the results, oral administration of C10-13 sodium linear alkylbenzene sulphonate (LAS-Na) to CRJ-SD rats at 0, 125, 250 and 500 mg/kg bw for 1 month resulted in a NOAEL of 125 mg/kg bw, based on reduced body weight, changes in organ weight (histopathology was not conducted) and clinical chemistry at the mid and high doses. The LOAEL was 250 mg/kg bw.
- Endpoint:
- sub-chronic toxicity: oral
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Justification for type of information:
- Read-across approach - see read-across justification in section 13.
- Reason / purpose for cross-reference:
- read-across source
- Principles of method if other than guideline:
- The purpose of this study is to determine the sub chronic toxicity of C10 - C14 linear alkylbenzenesulfonic 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 haematological 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 was performed.
- GLP compliance:
- no
- Remarks:
- Pre-dates GLP
- Limit test:
- no
- Species:
- rat
- Strain:
- Wistar
- Remarks:
- SLC
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- 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 - 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:
- unchanged (no vehicle)
- Remarks:
- 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)
- Dose / conc.:
- 40 mg/kg bw/day (nominal)
- Remarks:
- In diet (corresponding to 0.07% dose level)
- Dose / conc.:
- 115 mg/kg bw/day (nominal)
- Remarks:
- In diet (corresponding to 0.2% dose level)
- Dose / conc.:
- 340 mg/kg bw/day (nominal)
- Remarks:
- In diet (corresponding to 0.6% dose level)
- 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.
- 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
HAEMATOLOGY: 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 e
uthanasia (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 (all tissues weighed and also stomach, large and small intestines, pancreas, epidiymis, skin and bones) were examined and subjected to microscopy through H.E., PAS and silver staining. - 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.
- 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.
- 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:
- no effects observed
- Description (incidence and severity):
- No significant changes were observed in the animals.
- 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 animals did not recover fully by the end of the study. The animals moved slowly, and there 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.
HAEMATOLOGY:
- 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 and decrease in total protein.
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 swelling 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 thereon, 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% treated groups)
- Glomeruli contributed to overall swelling through dilation of vessel lumen and interstitial swelling (significant in in male/female rats of 0.2 and 0.07% dose groups. They were not very significant in the 1.8 and 0.6%)
- The glomerular interstitum had thawn mildly but chronically (significant for male/female rats in 1.8 and 0.6% groups)
- Cloudy swelling/vacuolar degeneration of proximal renal tubular epithelial cells were seen (slightly significant in LAS treated groups).
- Small, yellowish brown blobs the size of nuclei were 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
- The renal tubule had collapsed (significantly in male/females of 1.8% dose group)
- Swelling of surrounding connective tissue was observed in the interstitium (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. - 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 clin ical 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).
- 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
- 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:
The 26 weeks sub-chronic oral toxicity study of linear alkylbenzene sulfonic acid sodium salt (LAS) was performed in Wistar SLC rats. Approximately 4 weeks old male and femaleWistar SLC rats(obtained from Shizuoka Agricultural Cooperative Association for Laboratory Animals) with body weight range 80 - 90 g (males), 65 - 80 g (females)were used in the study. Five animals were housed in each stainless steelcage and maintained under controlled environmental conditions (temperature: Average of 25 ± 1°C, humidity:55 - 65%, and 12 hours light /12 hours dark). FII food (from Funabashi Farm)and tap water were provided ad libitum. The animals were administered daily with the LAS in diet at dose levels of0, 0.07, 0.2, 0.6 and 1.8%
(equivalent to 0,40, 115, 340, 1030 mg/kg bw/day) for 26 weeks. 10 animals/sex/dose group was taken in the study.
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 findings were obtained after euthanizing animals and organs were removed for organ weight measurements.Organ weights for brain, pituitary, thyroid, thymus, heart, lungs, liver, kidneys, spleen, adrenal gland, prostate, testes, uterus, ovaries and appendixwere recorded. Liver and kidney enzymes
were also analysed. Collected tissues were then examined and subjected to microscopy through H.E., PAS and silver staining. One male animal from the 1.8% dose group died. In the 1.8% dose groups, increase in relative weight of organs associated with diarrhea and significant suppression of weight gain has been remarked. There was also anemia and an increasing trend of white blood cell count, in addition to increased serum ALP activity, decrease in serum total protein and a very high level of tissue damage in the kidneys. In the 0.6% dose groups, there were mild suppression of weight gain, increased weight of appendix as well as fluctuations in serum ALP / total protein and tissue damage to the kidneys. In the 0.2% dose groups, there were increase in weight of appendix and histological abnormalities in a part of the kidneys. In the 0.2% dose groups, there were increase in weight of appendix and histological abnormalities in a part of the kidneys. 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. Administration of linear alkylbenzene sulfonic acid sodium salt (LAS) to Wistar SLC ratsat dose levels of 0, 0.07, 0.2, 0.6 and 1.8% (equivalent to 0,40, 115, 340, 1030mg/kg bw/day) for 26 weeks identified a NOAEL of 0.07% (40 mg/kg bw/day), based on tissue damage in caecum, kidney and liver, hematological as well as clinical chemistry parameters changes at higher dose levels (115 mg/kg bw/day and above).
- Endpoint:
- sub-chronic toxicity: oral
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- weight of evidence
- Justification for type of information:
- Read-across approach - see read-across justification in section 13.
- Reason / purpose for cross-reference:
- read-across source
- 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, hematological, 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
- Species:
- rat
- Strain:
- Wistar
- Remarks:
- 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 - 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:
- unchanged (no vehicle)
- Remarks:
- Test substance was administered either in diet or drinking water
- Analytical verification of doses or concentrations:
- no
- Duration of treatment / exposure:
- Nine months
- Frequency of treatment:
- Continuous in diet or drinking water (ad libitum)
- Dose / conc.:
- 300 mg/kg bw/day (nominal)
- Remarks:
- In diet (corresponding to 0.6% dose level)
- Dose / conc.:
- 900 mg/kg bw/day (nominal)
- Remarks:
- In diet (corresponding to 1.8% dose level)
- Dose / conc.:
- 0.07 other: %
- Remarks:
- In drinking water
- Dose / conc.:
- 0.2 other: %
- Remarks:
- In drinking water; 0.6 and 1.8% dose group (equivalent to 857.14 and 2571.43 mg/kg bw/day) were 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): 8 animals/sex/dose
Drinking water study: 9 animals/sex/dose - 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: 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), Hemoglobin (Hgb), Hematocrit (Hct), mean corpuscular volume (MCV) and mean corpuscular hemoglobin 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 - Clinical signs:
- effects observed, treatment-related
- Description (incidence and severity):
- Both female and male rats (drinking water study) 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 and 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):
- There were significant alteration in cholesterol [decrease; all doses, except female rats of 0.07% dose group (drinking water)], GPT [0.6% dose group (diet) females)], GOT [1.8% dose group (diet) males], albumin [1.8% (diet) males] , ALP levels [male and female rats fed with 1.8% LAS-diet] and cho
linesterase 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):
- 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. - Details on results:
- CLINICAL SIGNS: 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.
BODY WEIGHT AND WEIGHT CHANGES: There was significant decrease in body weight gain in male and females of 1.8% dose group fed with diet and in males of 0.2% dose group consuming LAS contained water.
WATER CONSUMPTION AND COMPOUND INTAKE: Increased water consumption was observed in males of 1.8% dose group fed with diet and 0.2% dose group consuming LAS contained water.
HAEMATOLOGICAL FINDINGS: 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: 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.
ORGAN WEIGHT: 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.
LIVER ENZYME TESTS: G6Pase activity was reduced in 1.8% dose group (diet) males and females, G6P-DH activity was reduced in 0.6 and 1.8% dose group (diet) males and females, where the percentage reduction was greater in 1.8% dose group (diet) animals. LDH activity was clearly reduced in 0.6, 1.8% dose group (diet), and 0.2% dose group (drinking water) males, but reduced in only 1.8% dose group (diet) females. GOT and GPT activities were clearly reduced in males, while in females GPT activity was reduced only in 1.8% dose group (diet) animals, and GOT activity was increased in 0.07 and 0.2% dose group (drinking water) animals but reduced in 1.8% dose group (diet) animals.
RENAL ENZYME TESTS: In males, Na, K-ATPase activity was significantly reduced in 0.2% dose group (drinking water) animals, and also reduced in other male treatment groups. A significant difference was also observed in G6Pase activity, where the reduction observed was associated with an increase in amount consumed. In females, G6Pase, Na, K-ATPase, and LDH activity were significantly reduced in 1.8% dose group (diet) animals, and G6Pase and LDH activity were also reduced in other treatment groups. - Dose descriptor:
- NOAEL
- Effect level:
- 85 mg/kg bw/day (nominal)
- Based on:
- act. ingr.
- Remarks:
- Drinking water
- 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 lactat e 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).
- Dose descriptor:
- LOAEL
- Effect level:
- 300 mg/kg bw/day (nominal)
- Based on:
- act. ingr.
- Remarks:
- Dietary 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
- 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
- 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
- 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 a 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 a 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:
The 9 months sub-chronic oral toxicity study of LAS was performed in Wistar JCL rats, focusing on the liver and kidneys.
Four week old male and female Wistar JCL rats (obtained from Shizuoka Agricultural Cooperative Association for Laboratory Animals) with body weight range100 - 124 g (males), 82 - 100 g (females) were used in the study. Five animals were housed in each cage and maintained under controlled environmental conditions (temperature: Average of 25 ± 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, 0.6 and 1.8% (equivalent to 0, 300 and 900 mg/kg bw/day); 8 animals/sex/dose
Dissolved in drinking water: 0, 0.07 and 0.2 % (equivalent to 0,85 and 145mg/kg bw/day); 9 animals/sex/dose
Rats in 0.6 and 1.8% dose group of 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 observed 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 appendixwere recorded. Liver and kidney enzymes were also analysed. No histopathology was performed.
No mortality was observed throughout the study. Both female and male rats consuming LAS-containing water exhibited a slight redness at the tips of their facial fur, and coarse fur over their bodies. There was significant decrease in body weight gain in male and females of 1.8% dose group fed with diet and in males of 0.2% dose group consuming LAS contained water. Increased water consumption was observed in males of 1.8% dose group fed with diet and 0.2% dose group consuming LAS contained water.
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. A marked reduction in cholesterol was observed in male and female rats of all dose groups [except female rats of 0.07% dose group (drinking water)] compared to controls. This indicate hepatocyte damage. 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) male 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.
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.
Liver enzymes were markedly reduced in 1.8% fed rats, due to impaired liver function, indicates reduced enzyme synthesis and direct enzyme inhibition by LAS or its metabolites. Renal G6Pase and Na, KATPase activity decreased, indicating kidney impairment.
Administration of LAS to Wistar JCL rats bytest 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 onsignificant 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).
Referenceopen allclose all
Table 1: Body weight gain, Food, water and sample consumption of Rats on administration of LAS for 9 months
| Dose group | No. of Rats | Initial BW (g) | Final BW (g) | BW gain (%) | Food (g/rat/day) | Water (g/rat/day) | Sample (g/kg bw/day) |
| 0 | 8M | 113.1±1.156 | 421.6±8.542 | 372.6±5.889 | 15 | 23 | - |
| 0.6 | 8M | 110.6±0.925 | 413.5±7.356 | 373.8±6.380 | 16 | 23 | 0.234 |
| 1.8 | 8M | 109.2±2.218 | 341.1±8.008 | 312.5±6.518** | 14 | 33 | 0.747 |
| 0.07 | 8M | 110.6±1.572 | 411.0±5.255 | 371.9±6.908 | 17 | 29 | 0.051 |
| 0.2 | 8M | 108.3±1.900 | 385.1±5.453 | 356.1±6.346** | 15 | 33 | 0.148 |
| 0 | 8F | 92.3±1.592 | 212.6±4.508 | 230.6±5.420 | 10 | 18 | - |
| 0.6 | 8F | 92.1±1.663 | 216.1±7.705 | 234.3±5.753 | 10 | 17 | 0.287 |
| 1.8 | 8F | 90.6±1.463 | 184.8±4.278 | 204.3±5.791** | 9 | 18 | 0.969 |
| 0.07 | 8F | 92.7±1.770 | 206.0±3.555 | 222.4±4.478 | 11 | 20 | 0.082 |
| 0.2 | 8F | 91.6±1.893 | 209.3±5.383 | 228.4±3.857 | 12 | 18 | 0.173 |
*p<0.05 **p<0.01
Table 2: Organ weight of male and female rats on administration of LAS for 9 months
| Dose group | Brain (g) / (g/100g) | Heart (g) / (g/100g) | Lung (g) / (g/100g) | Liver (g) / (g/100g) | Spleen (g) / (g/100g) | Kidner (R) (g) / (g/100g) | Kidney (L) (g) / (g/100g) | Adrenal (R) (mg) / (mg/100g) | Adrenal (L) (mg) / (mg/100g) | Testis/Ovary (R) (g) / (g/100g) | Testis/Ovary (L) (g) / (g/100g) | Uterus (g) / (g/100g) | |
| 0 M | 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±0.040 / 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 | - | |
| 0.6 M | 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 | - | |
| 1.8 M | 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** | - | |
| 0.07 M | 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 | - | |
| 0.2 M | 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 | - | |
| 0 F | 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±1.225 | 31.2±4.007 / 14.2±1.857 | 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 | 0.660±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 | 0.588±0.066 / 0.312±0.034 | |
| 0.07 F | 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±0.740 | 31.7±2.300 / 12.2±1.200 | 0.729±0.051 / 0.331±0.022 | |
| 0.2 F | 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 | 0.644±0.032 / 0.308±0.014 |
Table 1: body weight gain, food, water intake and feces excreation
| Sex | Group (%) | No. of animals | Initial BW (g) | Final BW (g) | BW gain (%) | Food (g)/animal/day | Water (g)/animal/day | Feces (g)/animal/day |
| Male | 0 | 10 | 85.00±0.87 | 387.00±9.50 | 452.10±9.8 | 14.17 | 16.90 | 2.31 |
| 0.07 | 10 | 80.90±1.92 | 387.10±5.93 | 480.74±13.11 | 14.48 | 16.91 | 2.24 | |
| 0.2 | 10 | 84.50±0.95 | 383.40±6.71 | 453.92±7.60 | 13.70 | 18.18 | 2.28 | |
| 0.6 | 10 | 86.40±1.54 | 365.80±8.51 | 426.23±8.89 | 13.84 | 16.28 | 2.23 | |
| 1.8 | 9/10 | 85.20±0.55 | 290.00±5.20 | 341.09±5.13** | 9.84 | 13.29 | 1.78 | |
| Female | 0 | 10 | 71.80±0.51 | 202.50±2.32 | 282.24±4.42 | 9.35 | 17.48 | 1.53 |
| 0.07 | 10 | 76.00±2.68 | 203.20±7.17 | 269.35±10.25 | 9.35 | 19.56 | 1.63 | |
| 0.2 | 10 | 74.30±1.11 | 201.10±3.33 | 271.05±5.31 | 8.90 | 17.69 | 1.39 | |
| 0.6 | 10 | 74.10±0.67 | 195.80±4.15 | 264.25±5.2* | 9.04 | 13.99 | 1.53 | |
| 1.8 | 10 | 74.30±0.94 | 158.80±3.41 | 214.02±5.29** | 7.01 | 12.14 | 1.23 |
* p<0.05 ** p<0.01
Table 2: Organ weight observations
| Group (%) | No. of animals | Spleen (mg) / (mg/100g) | Thymus (mg) / (mg/100g) | Liver (g) / (g/100g) | Lung (mg) / (mg/100g) | Kidney (R) (mg) / (mg/100g) | Kidney (L) (mg) / (mg/100g) | Hypophysis (mg/100g) | Heart (mg) / (mg/100g) |
| 0 | 10 M | 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 | 930±13 / 241±3 |
| 0.07 | 10 M | 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 | 932±16 / 240±2 |
| 0.2 | 10 M | 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 | 918±16 / 239±2 |
| 0.6 | 10 M | 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 | 878±18* / 235±4 |
| 1.8 | 9 M | 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 | 694±11** / 239±3 |
| 0 | 10 F | 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 | 596±10 / 265±6 |
| 0.07 | 10 F | 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 | 596±16 / 294±3 |
| 0.2 | 10 F | 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 | 574±13 / 258±5 |
| 0.9 | 10 F | 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* | 559±11* / 286±5 |
| 1.8 | 10 F | 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±0.63 | 426±11** / 269±7* |
* p<0.05 ** p<0.01
Table 2: Organ weight observations (continued..)
| Group (%) | No. of animals | Thyroid glands (mg) / (mg/100g) | Suprarenal gland (R) (mg) / (mg/100g) | Suprarenal gland (L) (mg) / (mg/100g) | Testis/Ovary (R) (mg) / (mg/100g) | Testis/Ovary (L) (mg) / (mg/100g) | Prosate/Uterus (mg) / (mg/100g) | Brain (mg) / (mg/100g) | Caecum (mg) / (mg/100g) |
| 0 | 10 M | 14.4±0.42 / 3.7±0.16 | 18.0±0.61 / 5.0±0.17 | 19.3±0.44 / 4.6±0.18 | 1592±20 / 412±7 | 1622±21 / 420±6 | 252±16 / 65±4.8 | 1983±12 / 515±11 | 837±34 / 216±8 |
| 0.07 | 10 M | 13.6±0.54 / 3.5±0.13 | 18.7±0.44 / 4.8±0.11 | 12.0±0.31 / 3.1±0.05 | 1643±14 / 425±6 | 1676±15 / 434±7 | 318±21* / 82±6.1 | 1918±106 / 494±25 | 938±38 / 242±9 |
| 0.2 | 10 M | 13.3±0.57 / 3.4±0.11 | 18.6±0.84 / 4.8±0.23 | 12.3±0.42 / 3.2±0.12 | 1621±20 / 423±4 | 1083±34 / 282±7 | 298±21 / 78±5.9 | 2020±11 / 528±8 | 971±35* / 253±7** |
| 0.6 | 10 M | 13.2±0.69 / 3.5±0.22 | 18.2±0.69 / 4.8±0.19 | 12.0±0.31 / 3.2±0.07 | 1612±36 / 432±11 | 1045±31 / 280±9 | 231±21 / 61±5.7 | 1969±18* / 528±12 | 939±41 / 251±11* |
| 1.8 | 9 M | 11.5±0.76** / 3.9±0.24 | 17.1±0.65* / 5.9±0.27* | 10.3±0.28** / 3.5±0.08** | 1501±36** / 518±13** | 836±19** / 288±3 | 231±21 / 79±7.0 | 1960±22 / 677±11** | 1484±30** / 512±12** |
| 0 | 9 F | 11.2±0.32 / 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 |
| 0.07 | 10 F | 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 |
| 0.2 | 10 F | 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 |
| 0.9 | 10 F | 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 |
| 1.8 | 10 F | 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** |
* p<0.05 ** p<0.01
Endpoint conclusion
- Endpoint conclusion:
- adverse effect observed
- Dose descriptor:
- NOAEL
- 85 mg/kg bw/day
- Study duration:
- subchronic
- Species:
- rat
- Quality of whole database:
- The database is considered sufficient to fulfil the specific requirement.
- Organ:
- kidney
Repeated dose toxicity: inhalation - systemic effects
Link to relevant study records
- Endpoint:
- sub-chronic toxicity: inhalation
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- Unavailable - orginal study report date 1988
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- comparable to guideline study with acceptable restrictions
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 413 (Subchronic Inhalation Toxicity: 90-Day Study)
- Deviations:
- yes
- Remarks:
- -whole body exposure; no urinalysis performed; the test substance characterization & stability data were not developed according to GLP.
- 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 Breeding Laboratory, Kingston, NY
- Housing: Individual suspended stainless steel cages over paper bedding. Animals receiving inhalation exposures were individually housed in all-wire cages.
- Diet: ad libitum
- Water: ad libitum
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21-23
- Humidity (%): 35-60
- Photoperiod (hrs dark / hrs light): 12/12 - Route of administration:
- inhalation: vapour
- Type of inhalation exposure:
- whole body
- Vehicle:
- air
- Remarks on MMAD:
- MMAD / GSD: not applicable
- Details on inhalation exposure:
- GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure chamber: Ten-cubic meter New York University style stainless steel chambers with pyramidal tops and bottoms
- Exposure apparatus: a pressurized tank through a capillary tube into a Laskin-style nebulizer located in the top of the chamber. The concentration of the test material in the inhalation chamber was controlled by regulating the pressure in the tank headspace, and consequently, the flow rate of the test material into the nebulizer.
- Method of holding animals in test chamber: the animals were positioned on the middle four rows of the cage racks and they were rotated weekly through the cage positions to ensure that all animals received similar exposure to the test material.
- Temperature, humidity, pressure in air chamber: monitored continuously and recorded approximately every 30 minutes
TEST ATMOSPHERE
- Brief description of analytical method used: Infrared spectroscopy
- Samples taken from breathing zone: no; the concentration in the chamber was routinely sampled five times per exposure at approximately one hour intervals.
VEHICLE (if applicable)
- Justification for use and choice of vehicle: no vehicle - Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- The test atmosphere was drawn through a MIRAN 1A General Purpose Gas Analyzer. The analytical method used was infrared spectroscopy.
- Duration of treatment / exposure:
- 13 weeks
- Frequency of treatment:
- 6 h/d, 5d/wk
- Dose / conc.:
- 20 mg/m³ air (analytical)
- Dose / conc.:
- 101 mg/m³ air (analytical)
- Dose / conc.:
- 499 mg/m³ air (analytical)
- No. of animals per sex per dose:
- 15
- Control animals:
- yes, concurrent no treatment
- Details on study design:
- - Dose selection rationale: not specified
- Observations and examinations performed and frequency:
- CAGE SIDE OBSERVATIONS: Yes, mortality and moribundity
- Time schedule: twice daily, but also during exposure period
BODY WEIGHT: Yes
- Time schedule for examinations: once per week
FOOD CONSUMPTION: No data
FOOD EFFICIENCY: No data
WATER CONSUMPTION: No data
OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: performed on all animals before exposures began, and on control and high level animals during the last exposure week
HAEMATOLOGY: Yes
- Time schedule for collection of blood: once during the study and once at the end of the study
- Anaesthetic used for blood collection: No data
- Animals fasted: Yes, food was withheld overnight before blood collection
- How many animals: 15/sex/dose level
- Parameters examined: Red blood cell count (RBC), white blood cell count (WBC), platelet count (PLT), hematocrit (Hct), level of hemoglobin (Hgb), and red blood cell indices [mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), and mean corpuscular hemoglobin concentration (MCHC)], leukocyte counts, reticulocyte count.
CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: once during the study and once at the end of the study
- Animals fasted: Yes
- How many animals: 15/sex/dose level
- Parameters examined: albumin, total protein, blood urea nitrogen (BUN), total bilirubin, glucose, glutamic pyruvic transaminase (D-GPT/ALT), alkaline phosphatase, glutamic oxaloacetate transaminase (D-GOT/AST), creatinine, cholesterol (Chol), calcium, phosphorus, chloride, sodium, and potassium. Globulin was determined by subtraction of albumin from the total protein value.
URINALYSIS: No
NEUROBEHAVIOURAL EXAMINATION: No - Sacrifice and pathology:
- GROSS PATHOLOGY: Yes, performed in all animals , organ weights determined (adrenals, brain, heart, kidneys, liver, spleen, testes with epididymides)
HISTOPATHOLOGY: Yes, all tissues were examined microscopically (aorta, adrenals, bone, brain, diaphragm, esophagus, eyes with optic nerve, gonads, heart, intestine, kidneys, liver, lung, lymph nodes, mammary gland, nasal passages, pancreas, pituitary, prostate, salivary gland, sciatic nerve, seminal vesicle, skeletal muscle, skin, spinal cord, spleen, stomach, thymus, thyroid, parathyroid, trachea, urinary bladder, uterus, vagina. - Statistics:
- Dunnett’s Multiple Comparison Test, Mann Whitney Test, Mann Whitney Test with Bonferroni Inequality Procedure, Fisher’s Exact Test with Bonferroni Inequality Procedure, Bartlett’s Test to evaluate homogeneity of variances, Analysis of Variance to determine if the sample (group) means could be considered as an estimate of a common population, and Grubb’s Test to detect outliers
- Clinical signs:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Not specified.
- Mortality:
- mortality observed, non-treatment-related
- Description (incidence):
- Two animals died during the study but it was not treatment related
- Body weight and weight changes:
- effects observed, treatment-related
- Description (incidence and severity):
- The mean body weight of high exposure level males was reduced (approximately 6-9%) throughout most of the study. Slight (approximately 4-6%) body weight reductions were also noted in high level females during weeks 6-13.
- Food consumption and compound intake (if feeding study):
- not examined
- Food efficiency:
- not examined
- Water consumption and compound intake (if drinking water study):
- not examined
- Ophthalmological findings:
- no effects observed
- Haematological findings:
- no effects observed
- Description (incidence and severity):
- minimal, non-dose related changes were seen in some hematological parameters (<3.5%) and some blood clinical parameters (<1.4%). The changes were not considered treatment related.
- Clinical biochemistry findings:
- effects observed, treatment-related
- Description (incidence and severity):
- At 500 mg/m3 a significant decrease in serum-glucose level of females was recorded.
- 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):
- Increased absolute and relative adrenal weights in high level females, probably treatment-related and may have been a non-specific response to stress; reduced spleen weights in high level males were attributed, at least partly, to the decreased body weights in the animals.
- Gross pathological findings:
- no effects observed
- Neuropathological findings:
- not examined
- Histopathological findings: non-neoplastic:
- effects observed, treatment-related
- Description (incidence and severity):
- Slight increases in centrilobular hepatocellular hypertrophy and individual hepatocellular necrosis in livers of high dose males, were not considered treatment-related. The mononuclear cell infiltrate occurred in kidneys of high dose females (small number) was considered spontaneous. At 500 mg/m3 inflammation of nasal mucosa was seen in female animals.
- Histopathological findings: neoplastic:
- no effects observed
- Key result
- Dose descriptor:
- NOAEC
- Effect level:
- 100 mg/m³ air
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: histopathology local effects (nose); decreased body weights and decreased serum glucose of high level females
- Critical effects observed:
- no
- Conclusions:
- Inflammation of the nasal mucosa decreased body weights and serum glucose were observed in females at 499 mg/m3. The NOAEC was therefore determined to be 100 mg/m3.
- Executive summary:
In a subchronic inhalation toxicity study (equivalent to OECD guideline 413), 2-propanamine (99.77% purity) was administered to 15 SD rats/sex/concentration via inhalation (dynamic whole body) exposure to concentrations of 0, 20, 101 or 499 mg/m³ for 6 hours per day, 5 days/week for a total of 13 weeks.
The results revealed a reduction in the average body weight of the male animals in the highly exposed group for most of the study period. The only significant change in haematology and clinical chemistry values, which was considered treatment-related, was a decrease in serum glucose (females in the high dose group). An increase in absolute and relative adrenal weights was observed in females in the high-dose group, probably treatment-related and may have been a non-specific response to stress; the decrease in spleen weights in males in the high-dose group was attributed, at least in part, to the decrease in body weight of the animals. No gross treatment-related pathological changes were detected, the only relevant microscopic change observed being inflammation of the nasal mucosa in the females tested at 499 mg/m3.The NOAEC is 100 mg/m3 based on histopathology local effects decreased body weights and serum glucose observed in females in the high-dose group.
This subchronic inhalation toxicity study in the rats is acceptable and satisfies the guideline requirement for a subchronic inhalation study OPPTS 870.3465; OECD 413 in the rats.
- Endpoint:
- sub-chronic toxicity: inhalation
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Justification for type of information:
- Read-across approach - see read-across justification in section 13.
- Reason / purpose for cross-reference:
- read-across source
- 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 Breeding Laboratory, Kingston, NY
- Housing: Individual suspended stainless steel cages over paper bedding. Animals receiving inhalation exposures were individually housed in all-wire cages.
- Diet: ad libitum
- Water: ad libitum
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21-23
- Humidity (%): 35-60
- Photoperiod (hrs dark / hrs light): 12/12 - Route of administration:
- inhalation: vapour
- Type of inhalation exposure:
- whole body
- Vehicle:
- air
- Remarks on MMAD:
- MMAD / GSD: not applicable
- Details on inhalation exposure:
- GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure chamber: Ten-cubic meter New York University style stainless steel chambers with pyramidal tops and bottoms
- Exposure apparatus: a pressurized tank through a capillary tube into a Laskin-style nebulizer located in the top of the chamber. The concentration of the test material in the inhalation chamber was controlled by regulating the pressure in the tank headspace, and consequently, the flow rate of the test material into the nebulizer.
- Method of holding animals in test chamber: the animals were positioned on the middle four rows of the cage racks and they were rotated weekly through the cage positions to ensure that all animals received similar exposure to the test material.
- Temperature, humidity, pressure in air chamber: monitored continuously and recorded approximately every 30 minutes
TEST ATMOSPHERE
- Brief description of analytical method used: Infrared spectroscopy
- Samples taken from breathing zone: no; the concentration in the chamber was routinely sampled five times per exposure at approximately one hour intervals.
VEHICLE (if applicable)
- Justification for use and choice of vehicle: no vehicle - Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- The test atmosphere was drawn through a MIRAN 1A General Purpose Gas Analyzer. The analytical method used was infrared spectroscopy.
- Duration of treatment / exposure:
- 13 weeks
- Frequency of treatment:
- 6 h/d, 5d/wk
- Dose / conc.:
- 20 mg/m³ air (analytical)
- Dose / conc.:
- 101 mg/m³ air (analytical)
- Dose / conc.:
- 499 mg/m³ air (analytical)
- No. of animals per sex per dose:
- 15
- Control animals:
- yes, concurrent no treatment
- Details on study design:
- - Dose selection rationale: not specified
- Observations and examinations performed and frequency:
- CAGE SIDE OBSERVATIONS: Yes, mortality and moribundity
- Time schedule: twice daily, but also during exposure period
BODY WEIGHT: Yes
- Time schedule for examinations: once per week
FOOD CONSUMPTION: No data
FOOD EFFICIENCY: No data
WATER CONSUMPTION: No data
OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: performed on all animals before exposures began, and on control and high level animals during the last exposure week
HAEMATOLOGY: Yes
- Time schedule for collection of blood: once during the study and once at the end of the study
- Anaesthetic used for blood collection: No data
- Animals fasted: Yes, food was withheld overnight before blood collection
- How many animals: 15/sex/dose level
- Parameters examined: Red blood cell count (RBC), white blood cell count (WBC), platelet count (PLT), hematocrit (Hct), level of hemoglobin (Hgb), and red blood cell indices [mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), and mean corpuscular hemoglobin concentration (MCHC)], leukocyte counts, reticulocyte count.
CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: once during the study and once at the end of the study
- Animals fasted: Yes
- How many animals: 15/sex/dose level
- Parameters examined: albumin, total protein, blood urea nitrogen (BUN), total bilirubin, glucose, glutamic pyruvic transaminase (D-GPT/ALT), alkaline phosphatase, glutamic oxaloacetate transaminase (D-GOT/AST), creatinine, cholesterol (Chol), calcium, phosphorus, chloride, sodium, and potassium. Globulin was determined by subtraction of albumin from the total protein value.
URINALYSIS: No
NEUROBEHAVIOURAL EXAMINATION: No - Sacrifice and pathology:
- GROSS PATHOLOGY: Yes, performed in all animals , organ weights determined (adrenals, brain, heart, kidneys, liver, spleen, testes with epididymides)
HISTOPATHOLOGY: Yes, all tissues were examined microscopically (aorta, adrenals, bone, brain, diaphragm, esophagus, eyes with optic nerve, gonads, heart, intestine, kidneys, liver, lung, lymph nodes, mammary gland, nasal passages, pancreas, pituitary, prostate, salivary gland, sciatic nerve, seminal vesicle, skeletal muscle, skin, spinal cord, spleen, stomach, thymus, thyroid, parathyroid, trachea, urinary bladder, uterus, vagina. - Statistics:
- Dunnett’s Multiple Comparison Test, Mann Whitney Test, Mann Whitney Test with Bonferroni Inequality Procedure, Fisher’s Exact Test with Bonferroni Inequality Procedure, Bartlett’s Test to evaluate homogeneity of variances, Analysis of Variance to determine if the sample (group) means could be considered as an estimate of a common population, and Grubb’s Test to detect outliers
- Clinical signs:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Not specified.
- Mortality:
- mortality observed, non-treatment-related
- Description (incidence):
- Two animals died during the study but it was not treatment related
- Body weight and weight changes:
- effects observed, treatment-related
- Description (incidence and severity):
- The mean body weight of high exposure level males was reduced (approximately 6-9%) throughout most of the study. Slight (approximately 4-6%) body weight reductions were also noted in high level females during weeks 6-13.
- Food consumption and compound intake (if feeding study):
- not examined
- Food efficiency:
- not examined
- Water consumption and compound intake (if drinking water study):
- not examined
- Ophthalmological findings:
- no effects observed
- Haematological findings:
- no effects observed
- Description (incidence and severity):
- minimal, non-dose related changes were seen in some hematological parameters (<3.5%) and some blood clinical parameters (<1.4%). The changes were not considered treatment related.
- Clinical biochemistry findings:
- effects observed, treatment-related
- Description (incidence and severity):
- At 500 mg/m3 a significant decrease in serum-glucose level of females was recorded.
- 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):
- Increased absolute and relative adrenal weights in high level females, probably treatment-related and may have been a non-specific response to stress; reduced spleen weights in high level males were attributed, at least partly, to the decreased body weights in the animals.
- Gross pathological findings:
- no effects observed
- Neuropathological findings:
- not examined
- Histopathological findings: non-neoplastic:
- effects observed, treatment-related
- Description (incidence and severity):
- Slight increases in centrilobular hepatocellular hypertrophy and individual hepatocellular necrosis in livers of high dose males, were not considered treatment-related. The mononuclear cell infiltrate occurred in kidneys of high dose females (small number) was considered spontaneous. At 500 mg/m3 inflammation of nasal mucosa was seen in female animals.
- Histopathological findings: neoplastic:
- no effects observed
- Key result
- Dose descriptor:
- NOAEC
- Effect level:
- 100 mg/m³ air
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: histopathology local effects (nose); decreased body weights and decreased serum glucose of high level females
- Critical effects observed:
- no
- Conclusions:
- Inflammation of the nasal mucosa decreased body weights and serum glucose were observed in females at 499 mg/m3. The NOAEC was therefore determined to be 100 mg/m3.
- Executive summary:
In a subchronic inhalation toxicity study (equivalent to OECD guideline 413), 2-propanamine (99.77% purity) was administered to 15 SD rats/sex/concentration via inhalation (dynamic whole body) exposure to concentrations of 0, 20, 101 or 499 mg/m³ for 6 hours per day, 5 days/week for a total of 13 weeks.
The results revealed a reduction in the average body weight of the male animals in the highly exposed group for most of the study period. The only significant change in haematology and clinical chemistry values, which was considered treatment-related, was a decrease in serum glucose (females in the high dose group). An increase in absolute and relative adrenal weights was observed in females in the high-dose group, probably treatment-related and may have been a non-specific response to stress; the decrease in spleen weights in males in the high-dose group was attributed, at least in part, to the decrease in body weight of the animals. No gross treatment-related pathological changes were detected, the only relevant microscopic change observed being inflammation of the nasal mucosa in the females tested at 499 mg/m3.The NOAEC is 100 mg/m3 based on histopathology local effects decreased body weights and serum glucose observed in females in the high-dose group.
This subchronic inhalation toxicity study in the rats is acceptable and satisfies the guideline requirement for a subchronic inhalation study OPPTS 870.3465; OECD 413 in the rats.
Referenceopen allclose all
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed
- Dose descriptor:
- NOAEC
- 100 mg/m³
- Study duration:
- subchronic
- Experimental exposure time per week (hours/week):
- 30
- Species:
- rat
- Quality of whole database:
- The database is considered sufficient to fulfil the specific requirement.
Repeated dose toxicity: inhalation - local effects
Link to relevant study records
- Endpoint:
- sub-chronic toxicity: inhalation
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Justification for type of information:
- Read-across approach - see read-across justification in section 13.
- Reason / purpose for cross-reference:
- read-across source
- 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 Breeding Laboratory, Kingston, NY
- Housing: Individual suspended stainless steel cages over paper bedding. Animals receiving inhalation exposures were individually housed in all-wire cages.
- Diet: ad libitum
- Water: ad libitum
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21-23
- Humidity (%): 35-60
- Photoperiod (hrs dark / hrs light): 12/12 - Route of administration:
- inhalation: vapour
- Type of inhalation exposure:
- whole body
- Vehicle:
- air
- Remarks on MMAD:
- MMAD / GSD: not applicable
- Details on inhalation exposure:
- GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure chamber: Ten-cubic meter New York University style stainless steel chambers with pyramidal tops and bottoms
- Exposure apparatus: a pressurized tank through a capillary tube into a Laskin-style nebulizer located in the top of the chamber. The concentration of the test material in the inhalation chamber was controlled by regulating the pressure in the tank headspace, and consequently, the flow rate of the test material into the nebulizer.
- Method of holding animals in test chamber: the animals were positioned on the middle four rows of the cage racks and they were rotated weekly through the cage positions to ensure that all animals received similar exposure to the test material.
- Temperature, humidity, pressure in air chamber: monitored continuously and recorded approximately every 30 minutes
TEST ATMOSPHERE
- Brief description of analytical method used: Infrared spectroscopy
- Samples taken from breathing zone: no; the concentration in the chamber was routinely sampled five times per exposure at approximately one hour intervals.
VEHICLE (if applicable)
- Justification for use and choice of vehicle: no vehicle - Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- The test atmosphere was drawn through a MIRAN 1A General Purpose Gas Analyzer. The analytical method used was infrared spectroscopy.
- Duration of treatment / exposure:
- 13 weeks
- Frequency of treatment:
- 6 h/d, 5d/wk
- Dose / conc.:
- 20 mg/m³ air (analytical)
- Dose / conc.:
- 101 mg/m³ air (analytical)
- Dose / conc.:
- 499 mg/m³ air (analytical)
- No. of animals per sex per dose:
- 15
- Control animals:
- yes, concurrent no treatment
- Details on study design:
- - Dose selection rationale: not specified
- Observations and examinations performed and frequency:
- CAGE SIDE OBSERVATIONS: Yes, mortality and moribundity
- Time schedule: twice daily, but also during exposure period
BODY WEIGHT: Yes
- Time schedule for examinations: once per week
FOOD CONSUMPTION: No data
FOOD EFFICIENCY: No data
WATER CONSUMPTION: No data
OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: performed on all animals before exposures began, and on control and high level animals during the last exposure week
HAEMATOLOGY: Yes
- Time schedule for collection of blood: once during the study and once at the end of the study
- Anaesthetic used for blood collection: No data
- Animals fasted: Yes, food was withheld overnight before blood collection
- How many animals: 15/sex/dose level
- Parameters examined: Red blood cell count (RBC), white blood cell count (WBC), platelet count (PLT), hematocrit (Hct), level of hemoglobin (Hgb), and red blood cell indices [mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), and mean corpuscular hemoglobin concentration (MCHC)], leukocyte counts, reticulocyte count.
CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: once during the study and once at the end of the study
- Animals fasted: Yes
- How many animals: 15/sex/dose level
- Parameters examined: albumin, total protein, blood urea nitrogen (BUN), total bilirubin, glucose, glutamic pyruvic transaminase (D-GPT/ALT), alkaline phosphatase, glutamic oxaloacetate transaminase (D-GOT/AST), creatinine, cholesterol (Chol), calcium, phosphorus, chloride, sodium, and potassium. Globulin was determined by subtraction of albumin from the total protein value.
URINALYSIS: No
NEUROBEHAVIOURAL EXAMINATION: No - Sacrifice and pathology:
- GROSS PATHOLOGY: Yes, performed in all animals , organ weights determined (adrenals, brain, heart, kidneys, liver, spleen, testes with epididymides)
HISTOPATHOLOGY: Yes, all tissues were examined microscopically (aorta, adrenals, bone, brain, diaphragm, esophagus, eyes with optic nerve, gonads, heart, intestine, kidneys, liver, lung, lymph nodes, mammary gland, nasal passages, pancreas, pituitary, prostate, salivary gland, sciatic nerve, seminal vesicle, skeletal muscle, skin, spinal cord, spleen, stomach, thymus, thyroid, parathyroid, trachea, urinary bladder, uterus, vagina. - Statistics:
- Dunnett’s Multiple Comparison Test, Mann Whitney Test, Mann Whitney Test with Bonferroni Inequality Procedure, Fisher’s Exact Test with Bonferroni Inequality Procedure, Bartlett’s Test to evaluate homogeneity of variances, Analysis of Variance to determine if the sample (group) means could be considered as an estimate of a common population, and Grubb’s Test to detect outliers
- Clinical signs:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Not specified.
- Mortality:
- mortality observed, non-treatment-related
- Description (incidence):
- Two animals died during the study but it was not treatment related
- Body weight and weight changes:
- effects observed, treatment-related
- Description (incidence and severity):
- The mean body weight of high exposure level males was reduced (approximately 6-9%) throughout most of the study. Slight (approximately 4-6%) body weight reductions were also noted in high level females during weeks 6-13.
- Food consumption and compound intake (if feeding study):
- not examined
- Food efficiency:
- not examined
- Water consumption and compound intake (if drinking water study):
- not examined
- Ophthalmological findings:
- no effects observed
- Haematological findings:
- no effects observed
- Description (incidence and severity):
- minimal, non-dose related changes were seen in some hematological parameters (<3.5%) and some blood clinical parameters (<1.4%). The changes were not considered treatment related.
- Clinical biochemistry findings:
- effects observed, treatment-related
- Description (incidence and severity):
- At 500 mg/m3 a significant decrease in serum-glucose level of females was recorded.
- 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):
- Increased absolute and relative adrenal weights in high level females, probably treatment-related and may have been a non-specific response to stress; reduced spleen weights in high level males were attributed, at least partly, to the decreased body weights in the animals.
- Gross pathological findings:
- no effects observed
- Neuropathological findings:
- not examined
- Histopathological findings: non-neoplastic:
- effects observed, treatment-related
- Description (incidence and severity):
- Slight increases in centrilobular hepatocellular hypertrophy and individual hepatocellular necrosis in livers of high dose males, were not considered treatment-related. The mononuclear cell infiltrate occurred in kidneys of high dose females (small number) was considered spontaneous. At 500 mg/m3 inflammation of nasal mucosa was seen in female animals.
- Histopathological findings: neoplastic:
- no effects observed
- Key result
- Dose descriptor:
- NOAEC
- Effect level:
- 100 mg/m³ air
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: histopathology local effects (nose); decreased body weights and decreased serum glucose of high level females
- Critical effects observed:
- no
- Conclusions:
- Inflammation of the nasal mucosa decreased body weights and serum glucose were observed in females at 499 mg/m3. The NOAEC was therefore determined to be 100 mg/m3.
- Executive summary:
In a subchronic inhalation toxicity study (equivalent to OECD guideline 413), 2-propanamine (99.77% purity) was administered to 15 SD rats/sex/concentration via inhalation (dynamic whole body) exposure to concentrations of 0, 20, 101 or 499 mg/m³ for 6 hours per day, 5 days/week for a total of 13 weeks.
The results revealed a reduction in the average body weight of the male animals in the highly exposed group for most of the study period. The only significant change in haematology and clinical chemistry values, which was considered treatment-related, was a decrease in serum glucose (females in the high dose group). An increase in absolute and relative adrenal weights was observed in females in the high-dose group, probably treatment-related and may have been a non-specific response to stress; the decrease in spleen weights in males in the high-dose group was attributed, at least in part, to the decrease in body weight of the animals. No gross treatment-related pathological changes were detected, the only relevant microscopic change observed being inflammation of the nasal mucosa in the females tested at 499 mg/m3.The NOAEC is 100 mg/m3 based on histopathology local effects decreased body weights and serum glucose observed in females in the high-dose group.
This subchronic inhalation toxicity study in the rats is acceptable and satisfies the guideline requirement for a subchronic inhalation study OPPTS 870.3465; OECD 413 in the rats.
- Endpoint:
- sub-chronic toxicity: inhalation
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- Unavailable - orginal study report date 1988
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- comparable to guideline study with acceptable restrictions
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 413 (Subchronic Inhalation Toxicity: 90-Day Study)
- Deviations:
- yes
- Remarks:
- -whole body exposure; no urinalysis performed; the test substance characterization & stability data were not developed according to GLP.
- 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 Breeding Laboratory, Kingston, NY
- Housing: Individual suspended stainless steel cages over paper bedding. Animals receiving inhalation exposures were individually housed in all-wire cages.
- Diet: ad libitum
- Water: ad libitum
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21-23
- Humidity (%): 35-60
- Photoperiod (hrs dark / hrs light): 12/12 - Route of administration:
- inhalation: vapour
- Type of inhalation exposure:
- whole body
- Vehicle:
- air
- Remarks on MMAD:
- MMAD / GSD: not applicable
- Details on inhalation exposure:
- GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure chamber: Ten-cubic meter New York University style stainless steel chambers with pyramidal tops and bottoms
- Exposure apparatus: a pressurized tank through a capillary tube into a Laskin-style nebulizer located in the top of the chamber. The concentration of the test material in the inhalation chamber was controlled by regulating the pressure in the tank headspace, and consequently, the flow rate of the test material into the nebulizer.
- Method of holding animals in test chamber: the animals were positioned on the middle four rows of the cage racks and they were rotated weekly through the cage positions to ensure that all animals received similar exposure to the test material.
- Temperature, humidity, pressure in air chamber: monitored continuously and recorded approximately every 30 minutes
TEST ATMOSPHERE
- Brief description of analytical method used: Infrared spectroscopy
- Samples taken from breathing zone: no; the concentration in the chamber was routinely sampled five times per exposure at approximately one hour intervals.
VEHICLE (if applicable)
- Justification for use and choice of vehicle: no vehicle - Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- The test atmosphere was drawn through a MIRAN 1A General Purpose Gas Analyzer. The analytical method used was infrared spectroscopy.
- Duration of treatment / exposure:
- 13 weeks
- Frequency of treatment:
- 6 h/d, 5d/wk
- Dose / conc.:
- 20 mg/m³ air (analytical)
- Dose / conc.:
- 101 mg/m³ air (analytical)
- Dose / conc.:
- 499 mg/m³ air (analytical)
- No. of animals per sex per dose:
- 15
- Control animals:
- yes, concurrent no treatment
- Details on study design:
- - Dose selection rationale: not specified
- Observations and examinations performed and frequency:
- CAGE SIDE OBSERVATIONS: Yes, mortality and moribundity
- Time schedule: twice daily, but also during exposure period
BODY WEIGHT: Yes
- Time schedule for examinations: once per week
FOOD CONSUMPTION: No data
FOOD EFFICIENCY: No data
WATER CONSUMPTION: No data
OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: performed on all animals before exposures began, and on control and high level animals during the last exposure week
HAEMATOLOGY: Yes
- Time schedule for collection of blood: once during the study and once at the end of the study
- Anaesthetic used for blood collection: No data
- Animals fasted: Yes, food was withheld overnight before blood collection
- How many animals: 15/sex/dose level
- Parameters examined: Red blood cell count (RBC), white blood cell count (WBC), platelet count (PLT), hematocrit (Hct), level of hemoglobin (Hgb), and red blood cell indices [mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), and mean corpuscular hemoglobin concentration (MCHC)], leukocyte counts, reticulocyte count.
CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: once during the study and once at the end of the study
- Animals fasted: Yes
- How many animals: 15/sex/dose level
- Parameters examined: albumin, total protein, blood urea nitrogen (BUN), total bilirubin, glucose, glutamic pyruvic transaminase (D-GPT/ALT), alkaline phosphatase, glutamic oxaloacetate transaminase (D-GOT/AST), creatinine, cholesterol (Chol), calcium, phosphorus, chloride, sodium, and potassium. Globulin was determined by subtraction of albumin from the total protein value.
URINALYSIS: No
NEUROBEHAVIOURAL EXAMINATION: No - Sacrifice and pathology:
- GROSS PATHOLOGY: Yes, performed in all animals , organ weights determined (adrenals, brain, heart, kidneys, liver, spleen, testes with epididymides)
HISTOPATHOLOGY: Yes, all tissues were examined microscopically (aorta, adrenals, bone, brain, diaphragm, esophagus, eyes with optic nerve, gonads, heart, intestine, kidneys, liver, lung, lymph nodes, mammary gland, nasal passages, pancreas, pituitary, prostate, salivary gland, sciatic nerve, seminal vesicle, skeletal muscle, skin, spinal cord, spleen, stomach, thymus, thyroid, parathyroid, trachea, urinary bladder, uterus, vagina. - Statistics:
- Dunnett’s Multiple Comparison Test, Mann Whitney Test, Mann Whitney Test with Bonferroni Inequality Procedure, Fisher’s Exact Test with Bonferroni Inequality Procedure, Bartlett’s Test to evaluate homogeneity of variances, Analysis of Variance to determine if the sample (group) means could be considered as an estimate of a common population, and Grubb’s Test to detect outliers
- Clinical signs:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Not specified.
- Mortality:
- mortality observed, non-treatment-related
- Description (incidence):
- Two animals died during the study but it was not treatment related
- Body weight and weight changes:
- effects observed, treatment-related
- Description (incidence and severity):
- The mean body weight of high exposure level males was reduced (approximately 6-9%) throughout most of the study. Slight (approximately 4-6%) body weight reductions were also noted in high level females during weeks 6-13.
- Food consumption and compound intake (if feeding study):
- not examined
- Food efficiency:
- not examined
- Water consumption and compound intake (if drinking water study):
- not examined
- Ophthalmological findings:
- no effects observed
- Haematological findings:
- no effects observed
- Description (incidence and severity):
- minimal, non-dose related changes were seen in some hematological parameters (<3.5%) and some blood clinical parameters (<1.4%). The changes were not considered treatment related.
- Clinical biochemistry findings:
- effects observed, treatment-related
- Description (incidence and severity):
- At 500 mg/m3 a significant decrease in serum-glucose level of females was recorded.
- 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):
- Increased absolute and relative adrenal weights in high level females, probably treatment-related and may have been a non-specific response to stress; reduced spleen weights in high level males were attributed, at least partly, to the decreased body weights in the animals.
- Gross pathological findings:
- no effects observed
- Neuropathological findings:
- not examined
- Histopathological findings: non-neoplastic:
- effects observed, treatment-related
- Description (incidence and severity):
- Slight increases in centrilobular hepatocellular hypertrophy and individual hepatocellular necrosis in livers of high dose males, were not considered treatment-related. The mononuclear cell infiltrate occurred in kidneys of high dose females (small number) was considered spontaneous. At 500 mg/m3 inflammation of nasal mucosa was seen in female animals.
- Histopathological findings: neoplastic:
- no effects observed
- Key result
- Dose descriptor:
- NOAEC
- Effect level:
- 100 mg/m³ air
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: histopathology local effects (nose); decreased body weights and decreased serum glucose of high level females
- Critical effects observed:
- no
- Conclusions:
- Inflammation of the nasal mucosa decreased body weights and serum glucose were observed in females at 499 mg/m3. The NOAEC was therefore determined to be 100 mg/m3.
- Executive summary:
In a subchronic inhalation toxicity study (equivalent to OECD guideline 413), 2-propanamine (99.77% purity) was administered to 15 SD rats/sex/concentration via inhalation (dynamic whole body) exposure to concentrations of 0, 20, 101 or 499 mg/m³ for 6 hours per day, 5 days/week for a total of 13 weeks.
The results revealed a reduction in the average body weight of the male animals in the highly exposed group for most of the study period. The only significant change in haematology and clinical chemistry values, which was considered treatment-related, was a decrease in serum glucose (females in the high dose group). An increase in absolute and relative adrenal weights was observed in females in the high-dose group, probably treatment-related and may have been a non-specific response to stress; the decrease in spleen weights in males in the high-dose group was attributed, at least in part, to the decrease in body weight of the animals. No gross treatment-related pathological changes were detected, the only relevant microscopic change observed being inflammation of the nasal mucosa in the females tested at 499 mg/m3.The NOAEC is 100 mg/m3 based on histopathology local effects decreased body weights and serum glucose observed in females in the high-dose group.
This subchronic inhalation toxicity study in the rats is acceptable and satisfies the guideline requirement for a subchronic inhalation study OPPTS 870.3465; OECD 413 in the rats.
Referenceopen allclose all
Endpoint conclusion
- Endpoint conclusion:
- adverse effect observed
- Dose descriptor:
- NOAEC
- 100 mg/m³
- Study duration:
- subchronic
- Species:
- rat
- Quality of whole database:
- Sufficient to address specific requirements.
Repeated dose toxicity: dermal - systemic effects
Endpoint conclusion
- Endpoint conclusion:
- no study available
Repeated dose toxicity: dermal - local effects
Endpoint conclusion
- Endpoint conclusion:
- no study available
Additional information
No information is available on the repeated dose toxicity of LAS IPA. The endpoint was addressed with data from LAS Na and IPA.
LAS Na:
Male and female rats were exposed to LAS Na (125, 250, 500 mg/kg bw/day) orally by gavage daily for 28 days. The results showed suppressed body weight gain, differences in some serum biochemical measures when compared to the controls, and decreased (spleen, heart, thymus) or increased (liver) organ weights in the animals of the highest dose level. The resultant LOAEL and NOAEL values were 250 and 125 mg/kg bw/day, respectively (Ito et al., 1978).
In a 6-month toxicity test male and female rats were exposed to LAS Na (CAS 69669-44-9) in the diet daily: 40, 115, 340, 1030 mg/kg bw/day. Diarrhea, suppressed growth, increased cecal weight, and degeneration of renal tubes characterized the highest dose group. Similar but less severe signs were seen in other doses with the exception of the lowest dose of 0.07%, which showed no adverse effects related to exposure to LAS. The resultant LOAEL and NOAEL values were 115 and 40 mg/kg bw/day, respectively (Yoneyama et al., 1972).
In a 9-month toxicity study male and female rats were exposed to LAS Na (CAS 69669-44-9; 85, 145, 430 mg/kg bw/day) in drinking water daily. Body weight was suppressed in the highest dose. Significant decreases in transaminase activity and renal Na, K-ATPase was seen in the second group. The resultant LOAEL and NOAEL values were 145 and 85 mg/kg bw/day, respectively (Yoneyama et al., 1976).
IPA:
Information for IPA are only available in this 90 -day inhalation study. Male and female Sprague-Dawley rats were dosed for 6 hrs/day, 5 days/wk for approximately 13 weeks with ispropylamine (IPA) vapours, at analytical exposure concentrations of 20, 101, and 499 mg IPA per cubic meter in air. The results revealed reduced mean body weights of male animals in the high exposure group throughout most of the study period. The only significant change in hematology and clinical chemistry values, which were considered treatment-related was a decrease in serum glucose (high dose group females). No treatment-related gross pathological changes were detected.The only relevant microscopic change observed was inflammation of the nasal mucosa in females tested with 499 mg/m3. The NOAEC of the study was set at 100 mg/m3.
Extrapolating the inhalatory NOAEC to an oral NOAEL with a sRV for rats of 0.29 mg/m3 (for a 6 -h exposure), and with a pulmonary absorption at least as great as absorption across the GI tract, the oral NOAEL for IPA will be 29 mg/kg bw. Extrapolation from the inhalation to the oral route is not likely to underestimate toxicity and therefore, it is considered acceptable.
The molecular conversion of this NOAEL to a NOAEL for LAS IPA leads to a value of 189 mg/kg bw, which is much higher than the 85 mg/kg bw proposed for LAS Na (94 mg/kg bw after molar scaling). Therefore, the NOAEL of 85 mg/kg bw/day (94 mg/kg bw after scaling) recorded in the 9-month feeding repeated-dose oral toxicity study for LAS-Na was selected, as the NOAEL for LAS IPA.
Justification for classification or non-classification
Based on the results of the repeated dose toxicity studies on read-across substances, LAS IPA does not warrant a STOT classification according to (CLP) Regulation (EC) No. 1272/2008.
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