Benzenesulfonic acid, 4-C15-16-sec-alkyl derivs.-, compd. with 1-aminopropane-2-ol

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

Key experimental studies exists for the read across substances Benzenesulfonic acid, C10-13-alkyl derivs., sodium salts and MIPA.

In the first key study the effects of exposure to Benzenesulfonic acid, C10-13-alkyl derivs., sodium salts were examined on reproductive toxicity. The test substance was given in the feed at doses of 0.02, 0.1, and 0.5% (14, 70, and 350 mg/kg bw d) for 84 days to four groups of weanling rats and evaluated for two years (three generations). Each dose consisting of 50 animals each of both sexes (P0-generation). When the P0-generation was 107-112 days old, 20 females from each dose group were mated with 20 males from the same group and maintained together for 17 days. The first litters of each generation (Fla- and F2a-generation) were sacrificed at 21 days of age. Ten days after the final litter was sacrificed, all females were remated with different males from the same group to obtain the F1b-generation. From the Flb-generation, 20 males and females of each group were selected at weaning to continue their respective diets and to be used for further reproduction studies. Reproduction studies on the F1b and F2b-generations were started when the rats were 80 to 85 days old, and were continued until the F3b-generation was weaned. All rats sacrificed at weaning were normal with respect to growth, organ to body weight ratios, gross pathology, and histology, and did not vary from controls. There were a number of statistically significant hematologic values, though these differences were small and did not indicate a trend or pattern. Overall, no significant effects were observed at the highest dose tested and the resulting NOAEL for the parental and both offspring generations was the highest dose tested, i.e., 350 mg/kg bw (0.5%).

In the second key study ( a combined repeated dose oral toxicity study with a reproduction/developmental screening study (OECD 422)), Wistar rats (12/sex/dose) were administered the hydrochloride salt of MIPA at 0, 100, 300 or 1000 mg/kg bw/day by oral gavage (BASF AG, 2008). The duration of treatment covered a 2-week premating period and mating period in both sexes, approximately 2 weeks post-mating in males, and the entire gestation period and 4 days of lactation in females, resulting in a total of 38 and 45 exposure days for males and females, respectively. Clinical pathology examinations revealed slightly, but statistically significant reduced hemoglobin and hematocrit values, which are indicative of a mild anemic process, in high dose males. Other (minor) clinical and pathological findings appear to be incidental and not dose-related. Based on the mild anemic process in males, the NOAEL for systemic toxicity is 300 mg/kg bw/day. No adverse effects were found on reproductive and fertility parameters, thus a NOAEL of 1000 mg/kg bw/day (the highest dose tested) was established for effects on fertility in males and females.

Link to relevant study records

Reference

Reference 1

Endpoint:

three-generation reproductive toxicity

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Principles of method if other than guideline:

Na-LAS (chain length distribution C10-14) was fed for 84 days to 4 groups of weanling rats (3 dose levels, plus control), each dose consisting of 50 animals each of both sexes (PO-generation). When the P0 generation was 107-112 days old, 20 females from each dose group were mated with 20 males from the same group and maintained together for 17 days. The first litters of each generation (Fla- and F2a-generation) were sacrificed at 21 days of age. Ten days after the final litter was sacrificed, all females were remated with different males from the same group to obtain the F1b generation. From the Flb-generation, 20 males and females of each group were selected at weaning to continue their respective diets and to be used for further reproduction studies. Reproduction studies on the F1b and F2b generations were started when the rats were 80 to 85 days old, and were continued until the F3b generation was weaned.

GLP compliance:

no

Limit test:

no

Species:

rat

Strain:

other: Charles River

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source:
- Age at study initiation: (P) weanling; (F1) 21 days
- Weight at study initiation: (P) Males: average 59.4-59.9 g; Females: average 57.0-57.3 g; (F1) Males: group weight 183.5-214.2 g; Females: group weight 157.8-193.2 g
- Housing: individual wire bottom cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°F): 76 +/- 3
- Humidity (%): 50 +/- 5
- Photoperiod (hrs dark / hrs light): 12/12 hrs

:

Route of administration:

oral: feed

Vehicle:

other: LAS was administered in feed (Purina Laboratory Meal) - no documentation of dilution prior to addition to meal

Details on mating procedure:

premating exposure period 84 days

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

2 years ( 3 generations)

Frequency of treatment:

continuous in feed

Details on study schedule:

- F1 parental animals not mated until 80-85 days old
- Selection of parents from F1 generation when pups were 21 days of age.
- Age at mating of the mated animals in the study: 107-112 days old

Remarks:

Doses / Concentrations:
0.02, 0.1, 0.5% (14, 70, 350 mg/kg bw d)
Basis:
actual ingested

No. of animals per sex per dose:

50 males and 50 females per group.

Control animals:

yes, concurrent no treatment

Litter observations:

Deformities and number of pups, average body weights, feed consumption, feed efficiency.

Postmortem examinations (parental animals):

Necropsy, body weight, organ to body weight ratios, routine hematology and histology.

Postmortem examinations (offspring):

Necropsy, body weight, organ to body weight ratios, routine hematology and histology.

Reproductive indices:

fertility, gestation, parturition, neonatal viability, lactation, and post-weaning growth

Clinical signs:

no effects observed

Body weight and weight changes:

no effects observed

Description (incidence and severity):

no effects to body weight were noted in the initial twelve weeks

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

no effects to body weight were noted in the initial twelve weeks

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Other effects:

no effects observed

Description (incidence and severity):

Test substance intake: no effects to average food consumption were noted in the initial twelve weeks

Reproductive function: oestrous cycle:

no effects observed

Reproductive function: sperm measures:

no effects observed

Reproductive performance:

no effects observed

No mortality or clinical signs were observed in parental animals. A statistically significant decrease in liver weights was noted in male rats at the low and mid dose levels at the 8 month sacrifice. As the decreased liver weight was within normal range, was not seen at the highest dose level, nor was seen at the 15 and 24 month sacrifices, it was not considered biologically significant. Body weight gains and organ to body weight ratios were normal. Gross examination revealed no abnormalities attributable to the test substance. General reproduction including fertility, gestation, parturition, neonatal viability, lactation, and post-weaning growth were normal for all test groups and did not vary from controls.

Key result

Dose descriptor:

NOAEL

Effect level:

350 mg/kg bw/day

Sex:

male/female

Basis for effect level:

other: 0.5%

Clinical signs:

not specified

Mortality / viability:

no mortality observed

Body weight and weight changes:

no effects observed

Sexual maturation:

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings:

no effects observed

Rats sacrificed at weaning were normal with respect to growth, organ to body weight ratios, gross pathology, and histology, and did not vary from controls. There were a number of statistically significant hematologic values, though these differences were small and did not indicate a trend or pattern.

Key result

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

350 mg/kg bw/day

Sex:

male/female

Basis for effect level:

other: 0.5%

Key result

Dose descriptor:

NOAEL

Generation:

F2

Effect level:

350 mg/kg bw/day

Sex:

male/female

Basis for effect level:

other: 0.5%

Key result

Reproductive effects observed:

not specified

Conclusions:

No significant effects on reproduction were observed at the highest concentration tested.

Executive summary:

Na-LAS was fed for 84 days to 4 groups of weanling rats for two years (three generations). No significant effects were observed at the highest dose tested and the resulting NOAEL for the parental and both offspring generations was 350 mg/kg bw (0.5%)

Reference 2

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study performed according to OECD Test Guideline 422 - Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

other: Wistar (Crl:WI(Han))

Sex:

male/female

Details on test animals or test system and environmental conditions:

- Source: Charles River Laboratories, Research Models and Services, Germany Gmbh
- Age at study initiation: 11-13 weeks
- Weight at study initiation: 267.4-309.0 g (males) and 186.7-217.7 g (females)
- Number of animals: 96 (12 per sex per dose group)
- Housing: Individually in type DK III stainless steel wire mesh cages supplied by Becker & Co., Castrop-Rauxel, Germany (floor area of about 800 cm2), with the following exceptions: for the overnight mating the females were put into the cages of the males; from day 18 p.c. until day 4 p.p. the pregnant animals and their litters were housed in Makrolon type M III cages (floor area about 800 cm2). The M III cages were also supplied by Becker & Co.. Pregnant females were provided with nesting material (cellulose wadding) toward the end of pregnancy.
- Diet (e.g. ad libitum): Ground Kliba maintenance diet mouse/rat ¿GLP¿ (Provimi Kliba SA, Kaiseraugst, Switzerland), ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24°C
- Humidity (%): 30-70%
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

other: tap water

Details on exposure:

- Dosing solution: pH 6.0 - 7.5
- Dose volume: 10 mL/kg bw

Details on mating procedure:

Mating Procedures:
- At least 13 days after the beginning of treatment, males and females from the same dose group were mated overnight in a ratio of 1:1
- The day of detection of sperm in the vaginal smear was designated day 0 of gestation
- Females were allowed to litter and rear their pups until day 4 after parturition

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

- The solutions were analyzed twice and determined to be 0, 1.61, 5.11, 15.73 g/100 mL using a content 68.9 g/100 g, which was determined by potentiometric titration and 0, 1.62, 4.67, 15.60 g/100 mL using 69.0 g/100 g, also determined by potentiometric titration

Duration of treatment / exposure:

38 days (males), 45 days (females)

Frequency of treatment:

daily

Details on study schedule:

Premating exposure period:
- Male: 13 days
- Female: 13 days

Remarks:

Doses / Concentrations:
0 (Vehicle), 100, 300 and 1000 mg/kg/day
Basis:
other: nominal

Remarks:

Doses / Concentrations:
0, 1.53, 4.59, and 15.29 g isopropanolamine HCl/100 mL assuming 65.4 g isopropanolamine HCl/100g
Basis:
other: calculated

No. of animals per sex per dose:

12

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: based on range finding study

Positive control:

None

Parental animals: Observations and examinations:

EXAMINATIONS:
- Mortality: twice daily (once on weekends and holidays) on all animals
- Clinical observations: once a day on all animals, detailed observations before test substance administration and weekly thereafter on all animals
- Food consumption and body weight: determined once per week with the following exceptions: not during mating for males or females and on day 0, 7, 14, and 20 post coitum for females and on days 0 and 4 postpartum for females with litter
- Body weight: Pups were weighed on days 1 and 4 postpartum.

Hematology and clinical chemistry: hematology (5 animals/sex/group) with EDTA-K3 as anticoagulant: Leukocytes, erythrocytes, hemoglobin, hematocrit, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, platelets, differential blood count, reticulocytes
- Prothrombin time (Hepato Quick's test)
- Clinical chemistry (5 animals/sex/group): alanine aminotransferase, aspartate aminotrasferase, alkaline phosphatase, gamma-glutamyltransferase, sodium, potassium, chloride, inorganic phosphate, calcium, urea, creatinine, glucose, total bilirubin, total protein, albumin, globulins, triglycerides, cholesterol, magnesium
- Urinalysis: on day 38 (males) and 45 (females) volume, color, turbidity, pH, protein, glucose, ketones, urobilinogen, bilirubin, blood, specific gravity, sediment
- Organs examined at necropsy (F0 animals only)
- Organ weight: liver, kidneys, adrenal glands, testes, epididymides, seminal vesicle, prostate, ovaries, uterus, thymus, spleen, brain, heart
- Microscopic: for all animals in the control and high dose group: trachea, lungs, liver, kidneys, spleen, adrenal glands, heart, brain, spinal cord (cervical, thoracic and lumbar), sciatic nerve, thyroid/parathyroid glands, testes, epididymides, ovaries, oviducts, uterus, vagina, prostate gland, seminal vesicles, coagulation glands, thymus, lymph nodes (mandibularr and mesenteric), stomach (forestomach and glandular stomach), duodenum, jejunum, ileum, cecum, colon, rectum, urinary bladder, bone marrow (femur)
- Microscopic: any organs which had gross pathological lesions at necropsy in any dose group
- Microscopic: for all male animals: liver
- Testes, epididymides and ovaries of animals that were killed as scheduled were fixed in Bouin's solution
- The number of implantation sites was determined by the method of Salewski, 1964 using uteri stained with 10% ammonium sulfide
- Functional observational battery and motor activity measurement: 5/group on day 36 for males and on day 43 for females
- Blood samples: from retroorbital venous plexus of 5 fasted F0 animals/group under isoflurane anesthesia on day 38 (males) and day 45 (females) followed by necropsy (including pups) under CO2 anesthesia

Litter observations:

Pup number and status at delivery: all pups delivered from the F0 parents were examined as soon as possible on the day of birth to determine the total number of pups and the number of liveborn and stillborn pups in each litter. Pups which died before the first determination of their status on the day of birth were defined as stillborn pups.
Pup viability and mortality: in general, a check was made for any dead or moribund pups twice daily on workdays (once in the morning and once in the afternoon) or as a rule, only in the morning on Saturdays, Sundays or public holidays. The number and percentage of dead pups on the day of birth (day 0 post partum) and pups dying between days 1 - 4 of the lactation period were determined; however, pups which died accidentally and pups which were sacrificed due to maternal death were not included in these calculations. The number of live pups/litter was calculated on the day of birth, and on lactation day 4. Furthermore the viability index was calculated.
Sex ratio: On the day of birth (day 0) the sex of the pups was determined by observing the distance between the anus and the base of the genital tubercle; normally, the anogenital distance is considerably greater in male than in female pups. The sex of the pups finally confirmed at necropsy. The sex ratio was calculated at day 0 and day 4 after birth.
Pup clinical observations: the live pups were examined daily for clinical symptoms (including gross-morphological findings) during the clinical inspection of the dams. If pups showed particular findings, these were documented with the dam concerned.

Postmortem examinations (offspring):

- Organs examined at necropsy (F1 pups): none, gross examination only, including those pups that did not survive to sacrifice

Statistics:

Food consumption (parental animals), body weight and body weight change (parental animals and pups; for the pup weights, the litter means were used), number of mating days, duration of gestation, number of pups delivered per litter, implantation sites, post implantation loss were evaluated by simultaneous comparison of all dose groups with the control group using the DUNNETT-test (two-sided) for the hypothesis of equal means.
Male and female mating index, male and female fertility index, gestation index, females with liveborn pups, females with stillborn pups, females with all stillborn pups, live birth index, pups stillborn, pups died, pups cannibalized, pups sacrificed moribund, viability index, number of litters with affected pups at necropsy were analyzed by pairwise comparison of each dose group with the control group using FISHER'S EXACT test for the hypothesis of equal proportions. Proportions of affected pups per litter with necropsy observations were analyzed by pairwise comparison of each dose group with the control group using the WILCOXON-test (one-sided) for the hypothesis of equal medians. Feces, rearing, grip strength of forelimbs and hindlimbs, landing footsplay test, motor activity were evaluated by non-parametric one-way analysis using KRUSKAL-WALLIS test (two-sided). If the resulting p-value was equal or less than 0.05, a pairwise comparison of each dose group with the control group was performed using Wilcoxon-test (two-sided) for the equal medians. Clinical pathology parameters, except reticulocytes and differential blood count were evaluated by non-parametric one-way analysis using KRUSKAL-WALLIS test (two-sided). If the resulting p-value was equal or less than 0.05, a pair-wise comparison of each dose group with the control group was performed using Wilcoxon-test (two-sided) for the equal medians. Urinalysis, except volume, color, turbidity and specific gravity was analyzed using pair-wise comparison of each dose group with the control group u

Reproductive indices:

Male mating index, male fertility index, female mating index, female fertility index, gestation index, post implantation loss

Offspring viability indices:

Live birth index

FUNCTIONAL OBSERVATIONAL BATTERY AND MOTOR ACTIVITY ASSESSMENT:
- No compound-related effects

FERTILITY/REPRODUCTIVE PERFORMANCE:
- No compound-related effects

F1 pups:
CLINICAL EXAMINATIONS (DAYS 0-4 POSTPARTUM):
- No adverse effects/findings

TOXIC EFFECTS BY DOSE LEVEL:
1000 mg/kg bw/day:
F0 parental animals:
CLINICAL EXAMINATIONS/CLINICAL PATHOLOGY/URINALYSIS/PATHOLOGY:
- Transient salivation in all males and most females after treatment from week 1 on
- Urine discoloration in all males and females for the entire study period
- Statistically significant decrease (p<= 0.01) in hemoglobin (-5%) and hematocrit (-6%) values in the peripheral blood of males
- Statistically significant increase in urea concentrations (+35%) in the serum of males
- Statistically significantly increase in albumin (+6%) in the serum of females
- Decreased (not significant) urine volume (-34%,-53%) with a subsequent increase (+2%,+4%) in specific gravity in males and females, respectively, without any changes in gross or microscopic parameters
- Statistically significant increase in absolute and relative liver weights in males with mild to minimal diffuse hepatocellular hypertrophy in 9/12 males, but no changes in liver enzymes, possibly adaptive
- Statistically significant increase in absolute and relative liver weights in females, without histological findings or changes in liver enzymes, but probably test substance-related and possibly adaptive
- Statistically significant increase in absolute and relative adrenal gland weights in males, not associated with any microscopic findings
- Statistically significant increase in absolute and relative thymus weights in females, not associated with any microscopic findings

300 mg/kg bw/day:
F0 parental animals
CLINICAL EXAMINATIONS/CLINICAL PATHOLOGY/URINALYSIS/PATHOLOGY:
- Transient salivation in all males after treatment from week 1 on.
- Statistically significant (p<=0.05) increase in hemoglobin (+6%) and hematocrit (+7%) values in the peripheral blood of females, not dose-related
- Statistically significant decrease in cholesterol in males, incidental
- Urine discoloration in all males and females from week 1 on.
- Statistically significant increase in body weight gain during premating in females, incidental
- Statistically significant increase in absolute and relative thymus weights in females, not associated with any microscopic findings
- Statistically significant increase in absolute and relative brain weights in females, incidental
- Statistically significant increase in relative kidney weights in females, incidental

100 mg/kg bw/day:
F0 parental animals
CLINICAL EXAMINATIONS/CLINICAL PATHOLOGY/URINALYSIS/PATHOLOGY:
- Statistically significant (p<=0.05) increase in hemoglobin (+6%) and hematocrit (+7%) values in the peripheral blood of females, not dose-related
- Statistically significant increase in body weight gain during premating in females, incidental

Dose descriptor:

NOAEL

Remarks:

for systemic toxicity

Effect level:

300 mg/kg bw/day

Sex:

male

Basis for effect level:

other: based on some indications of a mild anemic process

Dose descriptor:

NOAEL

Remarks:

for systemic toxicity

Effect level:

1 000 mg/kg bw/day

Sex:

female

Basis for effect level:

other: Maximum level tested

Dose descriptor:

NOAEL

Remarks:

for reproductive performance and fertility

Effect level:

1 000 mg/kg bw/day

Sex:

male/female

Basis for effect level:

other: Maximum level tested

Clinical signs:

no effects observed

Mortality / viability:

no mortality observed

Body weight and weight changes:

no effects observed

Sexual maturation:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

no effects observed

Histopathological findings:

not examined

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

1 000 mg/kg bw/day

Sex:

male/female

Basis for effect level:

other: Maximum level tested

Reproductive effects observed:

not specified

Absolute Weights	Male			Female
Group	1	2	3	1	2	3
Liver	+3%	-3%	+22%*	-4%	-4%	+17%*
Brain	-	-	-	0%	+4%*	-1%
Adrenal Glands	+4%	+4%	+19%*	-	-	-
Thymus	-	-	-	0%	+28%*	+26%*
*values were statistically significant different, P = 0.05

.

Relative Weights	Male			Female
Group	1	2	3	1	2	3
Liver	+3%	+1%	+23%*	-2%	0%	+16%*
Brain	-	-	-	+2%	+8%	-2%
Adrenal Glands	0%	+5%	+15%*	-	-	-
Thymus	-	-	-	+3%	+33%*	+25%**
Kidneys	-	-	-	+3%	+9%*	+4%
*values were statistically significant different, P <= 0.001, (**p= 0.05)

Conclusions:

- Only the findings of decreased hemoglobin and hematocrit at 1000 mg/kg bw/day in males are considered compound-related
- The other clinical and pathological findings appear to be incidental and not dose-related
- The NOAEL for reproductive performance and fertility is 1000 mg/kg bw/day for the F0 parental rats

- The NOAEL for reproductive performance and fertility is 1000 mg/kg body weight/day for the F0 parental rats
- The NOAEL for general, systemic toxicity is 300 mg/kg body weight/day for the F0 parental males based on some indications of a mild anemic process and 1000 mg/kg body weight/day for the F0 parental females
- The NOAEL for developmental toxicity in the F1 progeny is 1000 mg/kg body weight/day

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

350 mg/kg bw/day

Species:

rat

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

For reproductive toxicity, experimental studies are available for the read across substances Benzenesulfonic acid, C10-13-alkyl derivs., sodium salts and MIPA.

For Benzenesulfonic acid, C10-13-alkyl derivs., sodium salts, the potential for reproductive toxicity was examined in a three generation study in rats (two years). Overall, no significant effects were observed at the highest dose tested and the resulting NOAEL for the parental and both offspring generations was the highest dose tested, i.e., 350 mg/kg bw (0.5%).

For MIPA, in a combined repeated dose oral toxicity study with a reproduction/developmental screening study (performed according to OECD guideline 422), a NOAEL for reproductive performance and fertility of 1000 mg/kg bw/day (the highest dose tested) was established for parental males and females. Based on the mild anemic process in males, the NOAEL for systemic toxicity is 300 mg/kg bw/day.