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

Developmental toxicity / teratogenicity

Administrative data

Endpoint:

developmental toxicity

Type of information:

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

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: read across to Kilimisch 1 GLP, Guideline study (OECD 422)

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: OECD Test Guideline 422 - Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

1-aminopropan-2-ol hydrochloride

EC Number:

231-948-5

EC Name:

1-aminopropan-2-ol hydrochloride

Cas Number:

7780-04-3

IUPAC Name:

1-aminopropan-2-ol hydrochloride

Details on test material:

Hydrochloride salt of isopropanolamine (CAS no. 7780-04-3), 65.4 g isopropanolamine hydrochloride per 100 g aqueous solution (stored at room temperature).

Test animals

Species:

rat

Strain:

other: Wistar (Crl:WI(Han))

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

Administration / exposure

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

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

- These doses were calculated to be 0, 1.53, 4.59, 15.29 g isopropanolamine HCl/100 ml assuming 65.4 g isopropanolamine HCl/100g
- The solutions were analyzed twice and determined to be 0, 1.61, 5.11, 15.73 g/100ml using a content 68.9g/100g, which was determined by potentiometric titration and 0, 1.62, 4.67, 15.60 g/100 ml using 69.0 g/100g, also determined by potentiometric titration
- Dosing solution pH 6.0-7.5

Details on mating procedure:

- 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

Duration of treatment / exposure:

38 days (males), 45 days (females)

Frequency of treatment:

daily

Duration of test:

39-47 days

Doses / concentrations

Doses / concentrations 1

Remarks:

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

Doses / concentrations 2

Remarks:

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

Examinations

Maternal 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
- 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 males 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

Fetal examinations:

- Organs examined at necropsy (F1 pups): none, gross examination only (external and visceral findings), 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

Indices:

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

Results and discussion

Results: maternal animals

Maternal developmental toxicity

Details on maternal toxic effects:

Maternal toxic effects:yes

Details on maternal toxic effects:
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

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

FERTILITY/REPRODUCTIVE PERFORMANCE:
- No compound-related effects

Effect levels (maternal animals)

1

Dose descriptor:

NOAEL

Remarks:

for F0

Effect level:

1 000 mg/kg bw/day

Basis for effect level:

other: other:

2

Dose descriptor:

NOAEL

Remarks:

for F0 females

Effect level:

1 000 mg/kg bw/day

Basis for effect level:

other: other:

3

Dose descriptor:

NOAEL

Remarks:

for F0 males

Effect level:

300 mg/kg bw/day

Basis for effect level:

other: other:

4

Dose descriptor:

NOAEL

Remarks:

for F1

Effect level:

1 000 mg/kg bw/day

Basis for effect level:

other: developmental toxicity

Results (fetuses)

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
F1 pups:
- No adverse effects were observed in pup number, status at delivery, viability/mortality, sex ratio, clinical observations, body weight, and necropsy observations.

Effect levels (fetuses)

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day

Basis for effect level:

other: No adverse effects were observed in pup number, status at delivery, viability/mortality, sex ratio, clinical observations, body weight, and necropsy observations.

Fetal abnormalities

Abnormalities:

not specified

Overall developmental toxicity

Developmental effects observed:

not specified

Any other information on results incl. tables

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)

Applicant's summary and conclusion

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 developmental toxicity in the F1 progeny is 1000 mg/kg bw/day.

- 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