Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Description of key information

In a weight-of-evidence approach it is concluded, that MIPA does not induce effects on fertility: In a combined repeated dose oral toxicity study with a reproduction/developmental screening study (OECD422) in rats exposed to the hydrochloride salt of MIPA by gavage, a NOAEL for reproductive performance and fertility of 1000 mg/kg bw/day (equivalent to 673 mg/kg bw/day MIPA, the highest dose tested) was established for parental males and females. In an oral one-generation study with the structurally similar TIPA in rats according to FDA guidelines, the NOAEL for the parental generation as well as the off-spring was reported to be the highest dose tested 7500 ppm TIPA in males and females (equivalent to a 239 mg/kg bw/d intake of MIPA in males and 275 mg/kg bw/d in females).

Link to relevant study records

Referenceopen allclose all

Endpoint:
extended one-generation reproductive toxicity - basic test design (Cohorts 1A, and 1B without extension)
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
1988
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
ADAPTATION ACCORDING TO REACH ANNEX XI, section 1 - see justification attached to IUCLID section 13.2.
- Justification for WoE: RDT and Toxicity to reproduction
Qualifier:
according to
Guideline:
other: U.S. FDA Toxicological Principles for the Safety Assessment of Direct Food Additives and Color Additives Used in Food - 21 CFR 314.50(d)(2)
GLP compliance:
yes
Limit test:
no
Justification for study design:
A ccording to REACH Annex XI, section 1 and in line with the ECHA Guidance R.7a: Endpoint specific guidance (v6.0, July 2017) and R.6 QSARs and grouping of chemicals (May 2008), a testing does not appear scientifically necessary, because there is sufficient weight of evidence from available toxicological data for TIPA and structural similar substances (DIPA and MIPA), leading to the conclusion, that TIPA does not have an adverse effect on reproduction (fertility) and the available data are adequate to support a robust risk assessment.
- see justification attached to IUCLID section 13

SPECIFICATION OF STUDY DESIGN FOR ONE-GENERATION REPRODUCTION TOXICITY STUDY WITH JUSTIFICATIONS:

- Premating exposure duration for parental (P0) animals : 5 weeks
- Basis for dose level selection : selected after consultation with the FDA (for further details please refer to 'Details on study design' below)
- Exclusion of extension of Cohort 1B
- Termination time for F2 : no F2 generation was produced
- Exclusion of developmental neurotoxicity Cohorts 2A and 2B
- Exclusion of developmental immunotoxicity Cohort 3
- Route of administration : oral by feeding
Specific details on test material used for the study:
Purity: 98.76%
Species:
other:
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Breeding Laboratories, Kingston, New York
- Age: (P) 22 days old upon arrival and 42 days old at study start
- Weight at study initiation: (P) Males: 189.6-193.4 g; Females: 133.0-138.3 g
- Housing:
- during acclimation: 3 animals/cage, sexes separate, in stainless steel, wire-mesh cages suspended above Upjohn Deotized Animal Cage Boards (DACB) or R2 Reemay-backed cage boards
- after grouping: individually housed in stainless steel, wire-mesh cages suspended above Upjohn DACB or R2 Reemay-backed cage boards
- during the premating phase and throughout the study except during mating: housed on separate cage racks
- Diet: ad libitum, Irradiated Purina Certified Rodent Chow #5002 (IPCRC) diet
- Water: ad libitum, tap water
- Acclimation period: 14 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23±2
- Humidity (%): 50±10
- Air changes (per hr): not specified
- Photoperiod: 12-hour light/12-hour dark
IN-LIFE DATES: From: 03-09-1987 To: 09-08-1987
Route of administration:
oral: feed
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:

DIET PREPARATION
- TIPA was dried with Drierite drying agent under vacuum for several days. All compound was stored in containers which were purged with nitrogen after opening.
- Rate of preparation of diet: prepared weekly
- Mixing appropriate amounts with: distilled water (200 mL) and mixed on a magnetic stirrer until dissolved; added to Irradiated Purina Certified Rodent Chow (IPCRC) diet afterwards
- Storage temperature of food: refrigerated until used
Details on mating procedure:
- M/F ratio per cage: 1/1
- Length of cohabitation: maximum of seven days for the first approach; seven days for the second approach
- Proof of pregnancy: vaginal plug referred to as day 0 of pregnancy
- After 7 days of unsuccessful pairing replacement of first male by another male with proven fertility.
- Further matings after two unsuccessful attempts: no
- After successful mating each pregnant female was caged: individually in the polypropylene cages at the end of this period
Female rats were observed at least twice daily for signs of delivery starting several days prior to expected parturition.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Methanol was added to each diet sample and TIPA was extracted by sonication, filtered, evaporated under nitrogen, and derivatized with Pyridine and N-Trimethylsilylimidazole (TMSI). The reaction was completed by warming the solutions to 80°C for 10 (0, 500 ppm) or 20 minutes (2000, 7500 ppm). Recovery efficiencies were determined at the 500 ppm level by adding 5 mL of a 1000 µg/mL TIPA stock solution to control diet and at the 2000 and 7500 ppm levels by adding 20.3 and 75.9 mg H-16,648, respectively, to 10.0 g control diet. Extraction and preparation of the recovery samples were performed as described above. Calibration curves were created using standarrd solutions of TIPA or TEA. Peak area ratios were calculated (TIPA to TEA) and used for quantitation by linear regression analysis. The dietary concentration of TIPA was determined by multiple injection with a Hewlett-Packard 5880A gas chromatograph.
Duration of treatment / exposure:
The parental generation was fed TIPA in diet for five weeks.
After five weeks of feeding, they were mated to produce offspring which were fed diets containing TIPA for 90 days after weaning.
Frequency of treatment:
Daily
Dose / conc.:
500 ppm (nominal)
Remarks:
in diet (equivalent to a calculated 15.6 and 17.2 mg/kg bw/day intake of MIPA for males and females of the F1 generation, respectively)
Dose / conc.:
2 000 ppm (nominal)
Remarks:
in diet (equivalent to a calculated 63 and 71 mg/kg bw/day intake of MIPA for males and females of the F1 generation, respectively)
Dose / conc.:
7 500 ppm (nominal)
Remarks:
in diet (equivalent to calculated 239 and 275 mg/kg bw/day intake of MIPA for males and females of the F1 generation, respectively)
No. of animals per sex per dose:
25
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale:
Dose levels for this study were selected after consultation with the FDA. Dose levels were based upon acute and subchronic studies in rats conducted with TIPA in drinking water. On an acute basis, the single dose oral LD50 of TIPA in rats was 5,994 mg/kg bw. When rats were given 140 to 1,350 mg TIPA/kg bw/day in drinking water for 30 days, 1,350 mg/kg bw/day resulted in growth reduction and decreased food consumption. A dose level of 260 mg/kg bw/day produced unspecified histopathologic changes in some rats. No effects were seen at 140 mg/kg bw/day. In a 90-day study in rats, dose levels of 770 mg TIPA/kg bw/day in drinking water produced kidney effects consisting of dilation of Bowman's capsule and convoluted tubules, as well as marked cloudy swelling of liver parenchyma. A level of 220 mg/kg bw/day produced unspecified pathological changes in some animals. A concentration of 110 mg/kg bw/day for 90 days revealed no microscopic changes.
In the present study, the intermediate- and high-dose levels (2,000 and 7500 ppm in diet) were expected to produce mean daily intakes in rats of about 200 and 700 mg/kg bw/day, respectively. The intermediate-concentration level was selected to be similar to one at which pathological changes were seen in some rats on a previous study (220 mg/kg bw/day); the high-concentration level was selected to be comparable to a level at which pathological changes were seen in all rats (770 mg/kg bw/day).

- Fasting period before blood sampling for clinical biochemistry:
Two days prior to collection of blood samples for clinical evaluation, ten rats randomly selected from each group were placed in metabolism cages. The day before blood collection, these rats were fasted for approximately 16 hours. Urine was collected from each rat during this period. At the conclusion of this period, blood samples were collected from the orbital sinus of each rat while the rat was under light carbon dioxide anesthesia.
Positive control:
None
Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: conducted at least once daily throughout the study
- Cage-site examinations to detect moribund or dead rats and abnormal behavior and appearance among rats

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: during the premating phase of the study
- each rat was individually handled at each weighing and carefully examined for abnormal behavior and appearance

BODY WEIGHT: Yes
- Time schedule for examinations: once a week during the five-week premating phase of the study

FOOD CONSUMPTION AND COMPOUND INTAKE:
- Food consumption for each test group determined during the premating phase: Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes
Oestrous cyclicity (parental animals):
not examined
Sperm parameters (parental animals):
Parameters examined in F1 male parental generations:
- testis weight
Litter observations:
STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: yes
- If yes, maximum of 8 pups/litter (4/sex/litter as nearly as possible); excess pups were killed and discarded.

PARAMETERS EXAMINED
The following parameters were examined in F1 offspring:
- number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, weight gain, physical or behavioural abnormalities.

GROSS EXAMINATION OF DEAD PUPS:
- no
Postmortem examinations (parental animals):
None
Postmortem examinations (offspring):
SACRIFICE
- The F1 offspring were sacrificed at 111 (21 days p.p. weaning + 90 days feeding phase) days of age.
- These animals were subjected to postmortem examinations (macroscopic and/or microscopic examination) as follows:

HISTOPATHOLOGY / ORGAN WEIGTHS
- The following tissues indicated were prepared for microscopic examination and weighed, respectively:
Bone marrow, Lymph nodes (mandibular and mesenteric), Spleen, Aorta (thoracic), Heart, Salivary glands, Esophagus, Stomach, Liver (two sections), Pancreas, Small intestine (duodenum, jejunum, and ileum), Large intestine (cecum, colon, and rectum), Kidneys, Bladder, Pituitary, Thyroid - Parathyroid, Adrenals; Males: Prostate, Testes, Epididymides, Seminal vesicles, Females: Mammary gland, Ovaries, Uterus, Vagina; Brain (three coronal sections), Spinal cord, Peripheral nerve (sciatic), Bone (femur and sternum), Eyes, Exorbital lacrimal glands, Harderian glands, All gross lesions
Statistics:
For the P premating, gestation, and lactation, and F1 feeding phases, body weights, body weight gains, clinical laboratory measurements, and organ weights were analyzed by a one-way analysis of variance. When the test for differences among test group means (F test) was significant, pairwise comparisons between test and control groups were made with the Dunnett's test. Incidence of clinical observations was evaluated by the Fisher's Exact test with a Bonferroni correction and the Cochran-Armitage test for trend. Homogeneity of variances of organ weights and clinical laboratory data were analyzed with the Bartlett's test (alpha = 0.005). When the results of Bartlett's test were significant (variance was not homogeneous), the Mann-Whitney U test was employed instead of Dunnett's test for comparison of means (alpha = 0.05). Comparisons of offspring numbers and weights were made with the Mann-Whitney U test and Jonckheere's test for trend. Incidences of clinical observations in offspring were evaluated by Fisher's Exact test with a Bonferroni correction and the Cochran-Armitage test for trend. Indices of reproductive performance were also evaluated by Fisher's Exact test and the Cochran-Armitage test for trend.
Reproductive indices:
Further details given in other information.
Mating index, fertility index, gestation index
Offspring viability indices:
Further details given in other information.
Percent pups born alive, viability index, lactation index, litter survival, average number of pups/litter
Clinical signs:
no effects observed
Description (incidence and severity):
In the parent rats, there were no significant differences between groups in clinical observations during the premating period or in maternal rats during gestation or lactation.
Mortality:
no mortality observed
Description (incidence):
No rats died in the premating period.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
In the parent rats, there were no significant differences between groups in body weights and body weight gains during the premating period or in maternal rats during gestation or lactation.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
Food consumption and food efficiency of parental rats were similar between groups. Variability in the values over time and between sexes was related to the normal differences in body weight and food consumption relative to age and sex.
Food efficiency:
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
not examined
Other effects:
not examined
Reproductive function: oestrous cycle:
not specified
Reproductive function: sperm measures:
not specified
Reproductive performance:
no effects observed
Description (incidence and severity):
There were no differences attributed to administration of TIPA in gestation length, the number of litters produced (fertility index), or any other indices of reproductive performance.
Key result
Dose descriptor:
NOEL
Effect level:
7 500 ppm (nominal)
Based on:
test mat.
Remarks:
equivalent to 609 mg/kg bw/day for males; 700 mg/kg bw/day for females
Sex:
male/female
Remarks on result:
other: No treatment-related effects were observed up to the highest dose tested (602 mg/kg bw/day for males; 693 mg/kg bw/day for females)
Dose descriptor:
NOEL
Effect level:
7 500 ppm (nominal)
Based on:
test mat.
Remarks:
equivalent to 239 mg/kg bw/day MIPA for males; 275 mg/kg bw/day MIPA for females
Remarks on result:
other: No treatment-related effects were observed up to the highest dose tested (239 mg/kg bw/day MIPA for males; 700 mg/kg bw/day MIPA for females)
Key result
Critical effects observed:
no
Clinical signs:
no effects observed
Description (incidence and severity):
No treatment-related effects on F1 pups from any TIPA-treated groups were observed in clinical signs.
In the 90-day feeding phase of the F1 rats, there were no TIPA-related changes in clinical signs.
Mortality / viability:
no mortality observed
Description (incidence and severity):
No treatment-related effects on F1 pups from any TIPA-treated groups were observed in the number born or survival.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
No treatment-related, significant effects on F1 pups from any TIPA-treated groups were observed in body weights.
In the 90-day feeding phase of the F1 rats, there were no TIPA-related changes in body weights or body weight gains.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
In the 90-day feeding phase of the F1 rats, there were no TIPA-related changes in food consumption. The mean daily intake of TIPA over the entire 90-day feeding period was 0, 39.7, 160, and 609 mg/kg bw/day. For females at the same dose group the mean daily intake ws 0, 43.7,182, and 700 mg/kg bw/day over the same period.
Food efficiency:
not examined
Ophthalmological findings:
effects observed, non-treatment-related
Description (incidence and severity):
No changes in the ophthalmological examinations conducted on F1 rats were considered compound-related or biologically significant.
Haematological findings:
effects observed, non-treatment-related
Description (incidence and severity):
No changes in haematology on F1 rats was considered to be biologically significant or compound related.
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
No changes in the clinical chemistry evaluations conducted on F1 rats were considered compound-related or biologically significant.
Urinalysis findings:
effects observed, non-treatment-related
Description (incidence and severity):
No changes in the urine analyses conducted on F1 rats were considered compound-related or biologically significant.
Sexual maturation:
no effects observed
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Description (incidence and severity):
No changes in the organ weights conducted on F1 rats were considered compound-related or biologically significant.
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
No changes in the pathological evaluations conducted on F1 rats were considered compound-related or biologically significant.
Histopathological findings:
effects observed, non-treatment-related
Other effects:
not examined
Behaviour (functional findings):
not examined
Developmental immunotoxicity:
not examined
Key result
Dose descriptor:
NOEL
Generation:
F1
Effect level:
7 500 ppm (nominal)
Based on:
test mat.
Remarks:
(609 mg/kg bw/day for males; 700 mg/kg bw/day for females)
Sex:
male/female
Remarks on result:
other: No treatment-related effects were observed up to the highest dose tested (609 mg/kg bw/day for males; 700 mg/kg bw/day for females)
Dose descriptor:
NOEL
Generation:
F1
Effect level:
7 500 ppm (nominal)
Based on:
test mat.
Remarks:
(239 mg/kg bw/day MIPA for males; 275 mg/kg bw/day MIPA for females)
Sex:
male/female
Remarks on result:
other: No treatment-related effects were observed up to the highest dose tested (239 mg/kg bw/day MIPA for males; 275 mg/kg bw/day MIPA for females)
Key result
Critical effects observed:
no
Key result
Reproductive effects observed:
no

Table 1: Mean absolute organ weights (g) from male F1 rats

GROUP

CONC.(PPM)

LIVER

KIDNEYS

HEART

ADRENALS

BRAIN

TESTES

I-1

0

20.327 (2.457)

3.679 (0.388)

1.584 (0.120)

0.066 (0.011)

2.129 (0.191)

3.513 (0.231)

III-1

500

21.192 (2.372)

3.821 (0.478)

1.577 (0.148)

0.059 (0.012)

2.114 (0.133)

3.484 (0.231)

V-1

2000

22.022 (3.142)

3.900 (0.391)

1.688 (0.175)

0.066 (0.012)

2.200 (0.151)

3.617 (0.315)

VII-1

7500

21.743 (3.262)

3.924 (0.498)

1.626 (0.167)

0.067 (0.019)

2.181 (0. 143)

3.529 (0.417)

 

Table 2: Mean relative organ weights (% of body weight) from male F1 rats

GROUP

CONC.(PPM)

LIVER

KIDNEYS

HEART

ADRENALS

BRAIN

TESTES

I-1

0

3.7428 (.2441)

0.6792 (.0524)

0.2932 (.0235)

0.0122 (.0019)

0.3958 (.0501)

0.6518 (.0642)

III-1

500

3.8119 (.3235)

0.6874 (.0705)

0.2841 (.0237)

0.0106 (.0021)

0.3818 (.0326)

0.6284 (.0426)

V-1

2000

3.7627 (.3125)

0.6715 (.0746)

0.2898 (.0224)

0.0114 (.0021)

0.3794 (.0381)

0.6251 (.0822)

VII-1

7500

3.8580 (.3470)

0.6987 (.0640)

0.2902 (.0264)

0.0119 (.0035)

0.3902 (.0327)

0.6302 (.0722)

Table 3: Mean absolute organ weights (g) from female F1 rats

GROUP

CONC.

(PPM)

LIVER

KIDNEYS

HEART

ADRENALS

BRAIN

II-1

0

9.623(1.428)

2.212 (0.232)

1.023 (0.125)

0.076 (0.010)

1.995 (0.088)

IV-1

500

9.586(1.455)

2.167 (0.266)

1.029(0.112)

0.073 (0.012)

1.984 (0.064)

VI-1

2000

9.979(1.485)

2.301 (0.201)

1.073 (0.097)

0.078 (0.014)

1.969 (0.075)

VIII-1

7500

9.596(1.242)

2.255 (0.216)

1.027 (O.097)

0.075 (0.014)

1.930 (0.185)

Table 4: Mean absolute organ weights (% of body weight) from female F1 rats

GROUP

CONC.

(PPM)

LIVER

KIDNEYS

HEART

ADRENALS

BRAIN

II-1

0

3.3253 (.2382)

0.7685 (.0638)

0.3552 (.0349)

0.0265 (.0048)

0.6982(.0758)

IV-1

500

3.3637(.2409)

0.7627 (.0521)

0.3631 (.0300)

0.0256 (.0031)

0.7051 (.0715)

VI-1

2000

3.2420(.3182)

0.7539 (.0889)

0.3503 (.0284)

0.0255 (.0045)

0.6458(.0654)

VIII-1

7500

3.4011 (.2820)

0.8009 (.0562)

0.3659 (.0381)

0.0269 (.0053)

0.6888 (.0824)

Standard deviation in paranthesis

+ - Significantly different (P<0.05) from control group by LSD (least standard deviation)

# - Significantly different (P<0.05) from control group LSD and Dunnet's test

Conclusions:
The two highest levels produced mean daily intake comparable to those at which pathological changes of the kidneys and liver were reported in a drinking water study by another laboratory. However, in the present study no effects were seen at 7,500 ppm. The no-observable-effect level (NOEL) for this study was 7,500 ppm since no effects were observed at any dietary concentration in parental rats or in offspring rats prior to or after weaning.
Executive summary:

The reported feeding study in Crl:CDBR rats describes a combination of a sub-chronic and in utero study, i.e. F1 offspring rats were fed triisopropanolamine (TIPA) for 90-days after exposure prior to weaning / during pregnancy.

Prior to weaning, these F1 rats were exposed to TIPA in utero in maternal milk during lactation, and in any diet they consumed. The P rats had been continuously fed their group's assigned concentration of TIPA for five weeks prior to mating and during mating, gestation and lactation. Twenty-five rats/sex/group were used for the P generation. Among offspring of the P rats, twenty/sex/group were selected to continue as the F1 generation. Both generations were fed diets that contained 0, 500, 2,000, or 7,500 ppm of TIPA.

Clinical signs, mortality, and body weights of parental rats were recorded throughout the premating period and for females continued to be recorded during the gestation and lactation periods. Food consumption and mean daily intake of TIPA were determined during the premating period for both sexes and for females during gestation. After production of the F1 generation was complete, P rats were sacrificed and discarded without pathological examination. Clinical signs, counts, mortality, and group body weights of F1 offspring were recorded prior to weaning.

At weaning, 20 randomly selected F1 rats/sex/group were selected to continue on their respective treatment group's diet (one/sex/litter when possible). The remainder were sacrificed and discarded without pathological examination. Clinical observations, mortality, body weights, and food consumption of the F1 offspring were recorded during a 90-day feeding phase. F1 offspring were given ophthalmological examinations at the beginning and end of the 90-day feeding period. Clinical chemistry and urine analyses were conducted after approximately 45 and 90 days of feeding.

At the end of the 90-day feeding period, all surviving rats were sacrificed and given a gross pathological examination. Tissues from the control and high concentration groups were examined microscopically. Lungs, liver, kidneys, and organs with lesions in the low- and intermediate-concentration groups were also examined microscopically.

In the parent rats, there were no significant differences between groups in clinical observations, body weights, and weight gains during the premating period or in maternal rats during gestation or lactation. No parental rats died during the premating, mating, gestation, or lactation period. Food consumption and food efficiency of parental rats were similar between groups.

There were no differences attributed to administration of TIPA in gestation length, the number of litters produced (fertility index), or any other indices of reproductive performance.

No treatment-related effects on F1 pups from any TIPA-treated groups were observed in the number born, survival, body weights, or clinical signs.In the 90-day feeding phase of the F1 rats, there were no TIPA-related changes in clinical signs, body weights, body weight gains, or food consumption. One death occurred in the high-concentration group, but is not considered compound related. The mean daily intake of TIPA over the entire

90-day feeding period was 0, 39.7, 160, and 609 mg/kg/day (equivalent to a 15.6, 63 and 239 mg/kg bw/d intake of MIPA) for the control, low-, intermediate-, and high-concentration male groups, respectively. For females at the same concentrations the mean daily intake was 0, 43.7, 182, and 700 mg/kg/day (equivalent to a 17.2, 71 and 275 mg/kg bw/d intake of MIPA) over the same period. No changes in the clinical chemistry evaluations, urine analyses, ophthalmological examinations, organ weights, or pathological evaluations conducted on F1 rats were considered compound-related or biologically significant.

The no-observable-effect-level (NOEL) on this study was 7,500 ppm, the highest level tested.

Endpoint:
screening for reproductive / developmental toxicity
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
ADAPTATION ACCORDING TO REACH ANNEX XI, section 1 - see justification attached to IUCLID section 13.2.
- Justification for WoE: RDT and Toxicity to reproduction
Reason / purpose:
reference to same study
Reason / purpose:
reference to same study
Qualifier:
according to
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Version / remarks:
22 March 1996
Deviations:
no
GLP compliance:
yes
Limit test:
no
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and batch No.of test material: BASF and BA 1449
- Purity test date: 04 Jan 2006
- concetration of given stock solution:65.4 g isopropanolamine hydrochloride per 100 g aqueous solution

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature (freezer for reanalysis)
- Stability under test conditions: confirmed by reanalysis
- Solubility and stability of the test substance in the solvent/vehicle: stability of aq. solution confirmed by reanalysis

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: test material was weighed up and dissolved in tap water

FORM AS APPLIED IN THE TEST: solution
Species:
rat
Strain:
Wistar
Remarks:
Crl:WI(Han)
Sex:
male/female
Details on test animals 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
- Diet: Ground Kliba maintenance diet mouse/rat “GLP” (Provimi Kliba SA, Kaiseraugst, Switzerland), ad libitum
- Water: ad libitum from drinking bottles
- 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:
water
Remarks:
tap water
Details on exposure:
TEST ITEM
- Dosing solution: pH 6.0 - 7.5

VEHICLE
- Amount of vehicle: 10 mL/kg bw
Details on mating procedure:
- M/F ratio per cage: 1:1
- Length of cohabitation: maximum of 2 weeks
- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of pregnancy
- After successful mating each pregnant female was caged: in Makrolon type M III cages (floor area about 800 cm2)from day 18 p.c. until day 4 p.p., supplied with nesting material (cellulose wadding) toward the end of pregnancy
- 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
Dose / conc.:
100 mg/kg bw/day (nominal)
Remarks:
equivalent to 67 mg/kg bw/day Isopropanolamine
Dose / conc.:
300 mg/kg bw/day (nominal)
Remarks:
equivalent to 202 mg/kg bw/day Isopropanolamine
Dose / conc.:
1 000 mg/kg bw/day (nominal)
Remarks:
equivalent to 673 mg/kg bw/day Isopropanolamine
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:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily (littering and lactation behaviour evaluated in the mornings
- Cage side observations included any signs of morbidity, pertinent behavioral changes and signs of overt toxicity, documented for each animal

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: prior to the administration period and thereafter at weekly intervals
- Parameters: abnormal behavior during “handling”, fur, skin, salivation, nose discharge, lacrimation, pupil size, posture, palpebral closure, respiration, tremors, convulsions, abnormal movements, impairment of gait, activity/arousal level, feces (appearance/consistency), urine, other findings

BODY WEIGHT: Yes
- Time schedule for examinations:once a week at the same time of the day (in the morning);
- Time schedule exceptions: during the mating period the parental females were weighed on the day of positive evidence day 0 p.c. and on days 7, 14 and 20 p.c.; females with litter: day day 0 p.p. and on day 4 p.p.; females without a litter: weekly

FOOD CONSUMPTION AND COMPOUND INTAKE: Yes
- once a week (in a period of 7 days) for male and female parental animals
- exceptions: not determined during the mating period; F0 females with evidence of sperm were determined on days 0, 7, 14 and 20 p.c.; F0 females, which gave birth to a litter were determined on days 0 and 4 p.c.
- not determined in females without positive evidence of sperm and females without litter

HAEMATOLOGY 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

URINANALYSIS:
- on day 38 (males) and 45 (females) volume, color, turbidity, pH, protein, glucose, ketones, urobilinogen, bilirubin, blood, specific gravity, sediment
- Parameters: volume, color, turbidity, pH, protein, glucose, ketones, urobilinogen, bilirubin, blood, specific gravity (urine refractometer), sediment (microscopically)

Functional observation battery (FOB) and motor activity measurement:
- on study day 36 in first 5 male/group and on study day 43 in first 5 female/group
Sperm parameters (parental animals):
Parameters examined in F0 male parental generations: testes weight, epididymis weight
Litter observations:
STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: no

PARAMETERS EXAMINED
The following parameters were examined in F1 offspring: number and sex of pups, stillbirths, live births, postnatal mortality, weight gain (PND 1-4), presence of gross anomalies, anogenital distance (AGD)

GROSS EXAMINATION OF DEAD PUPS: yes, for external and internal abnormalities

ASSESSMENT OF DEVELOPMENTAL NEUROTOXICITY: no

ASSESSMENT OF DEVELOPMENTAL IMMUNOTOXICITY: no
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: after blood from 5 F0 males per group was sampled, on study day 38 necropsy of all male animals was performed
- Female animals: after blood from 5 F0 females per group was sampled, on study day 45 necropsy of all female animals was performed

GROSS NECROPSY
- Gross necropsy was not further specified.

HISTOPATHOLOGY / ORGAN WEIGHTS
- The tissues indicated in Table 1 were prepared for microscopic examination.
- The animals/organs were weighed: anesthetized animals, liver, kidneys, adrenal glands, testes, epididymides, seminal vesicle, prostate, ovaries, uterus, thymus, spleen, brain, heart
Postmortem examinations (offspring):
- Organs examined at necropsy (F1 pups): all surviving pups, all stillborn pups and those pups that died before schedule

SACRIFICE
- The F1 offsprings were sacrificed on day 4 post partum.
- These animals were subjected to postmortem examinations (macroscopic and/or microscopic examination) as follows:

GROSS NECROPSY
- Gross necropsy consisted of external and internal (eviscerated) examinations including and their organs were assessed macroscopically.
Statistics:
Due to limitations of characters, details for statistics are given as tables (2, 3, 4) under part 'other information'.

CLINICAL EXAMINATIONS
- DUNNETT-test (two-sided) for the hypothesis of equal means
- Pairwise comparison of each dose group with the control group using FISHER'S EXACT test for the hypothesis of equal proportions
- Pairwise comparison of each dose group with the control group using the WILCOXON-test (onesided) for the hypothesis of equal medians
- 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 (twosided) for the equal medians

CLINICAL PATHOLOGY
- 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
- Pair-wise comparison of each dose group with the control group using FISHER's exact test for the hypothesis of equal proportions

HISTOPATHOLOGY
- Means and standard deviations AND KRUSKAL-WALLIS and WILCOXON test
Reproductive indices:
MALE MATING AND FERTILITY INDICES

Male mating index (%) = (number of males with confirmed mating* / number of males placed with females) x 100
* defined by a female with vaginal sperm or with implants in utero

Male fertility index (%) = (number of males proving their fertility* / number of males placed with females) x 100
* defined by a female with implants in utero


FEMALE REPRODUCTION AND DELIVERY DATA

Female mating index (%) = (number of females mated* / number of females placed with males) x 100
* defined as the number of females with vaginal sperm or with implants in utero

Female fertility index (%) = (number of females pregnant* / number of females mated**) x 100
* defined as the number of females with implants in utero
** defined as the number of females with vaginal sperm or with implants in utero

Gestation index (%) = (number of females with live pups on the day of birth / number of females pregnant*) x100
* defined as the number of females with implants in utero

Live birth index (%) = (number of liveborn pups at birth / total number of pups born) x 100

Post implantation loss (%) = ([number of implantations - number of pups delivered] / number of implantations) x 100
Offspring viability indices:
Viability index (%) = (number of live pups on day 4 after birth / number of liveborn pups on the day of birth) x 100

Sex ratio = (number of live male or female pups on day 0/4 / number of live male and female pups on day 0/4) x 100
Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
- no test substance-related or spontaneous clinical findings observed in the male and female animals of 100 mg/kg body weight/day test group
- salivation after treatment was seen in all high dose male animals (1000 mg/kg body weight/day) during study weeks 1-5, finding persisted in the respective males only for a few minutes after daily gavage dosing
- urine discoloration was observed in all male animals of the high dose group during the whole study period (week 0-5) and in all mid dose male animals (300 mg/kg body weight/day) during study weeks 1-5 and females, for all high dose females during the whole study period (week 0 - 6) and in all mid dose females during study weeks 1-6
- 7 out of 12 female animals of test group 3 showed salivation after treatment during study weeks 1-6, finding persisted in the respective females only for a few minutes after daily gavage dosing
- detailed clinical observations on study days 0, 7, 14, 21, 28, 35 and additionally day 42 for female animals did not reveal any additional, substance-induced abnormalities in the male and female animals of the test groups 0-3 (0, 100, 300 and 1000 mg/kg body weight/day)

The clinical findings, i.e. transient salivation and discolored urine are considered to be substance-induced, but are without any toxicological relevance and are not assessed as being adverse.
It is assumed that the temporary salivation was induced by the bad taste of the test substance (small droplets of the test substance solution might be adjacent on the tip of the gavage) or minor local affection of the upper digestive tract (without showing a morphological correlate at pathology). The urine discoloration is possibly related to a chemical reaction of the test substance or its metabolites with the bedding or with components of the air.
Mortality:
no mortality observed
Description (incidence):
There were no substance-related or spontaneous mortalities in any of the groups.
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
Body weights and body weight gains of all male and female (+ during the gestation, lactation period and after weaning) animals of all substance-treated groups (100, 300 and 1000 mg/kg body weight/day) were comparable to that of the concurrent control group during the whole study taking normal biological variability into account.
The statistically significantly increased body weight gains of the low and mid dose females (100 and 300 mg/kg body weight/day) during premating weeks 1-2 are considered to be spontaneous in nature and not to be adverse.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
There were no effects observed for food consumption compared to the control group.
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
- slightly, but statistically significantly decreased hemoglobin and hematocrit values in the peripheral blood of high dose males
- no treatment related effects were seen in the other hematology parameters of males and in the hematology investigations of females
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
- differences in serum enzyme activities were not evident at any dose level in either males or females
- blood chemistry results showed statistically significantly increased urea concentrations in the serum of high dose males and significantly higher albumin levels in the serum of high dose females
- marginally increased urea concentrations were also seen in the high dose females
- the increase in females was not sufficient to be statistically identified
- no treatment related changes were found in the other blood chemistry parameters
Urinalysis findings:
effects observed, non-treatment-related
Description (incidence and severity):
- decreased amounts of urine with increased specific gravity in all animals, more pronounced in females than in males
- no treatment related effects were seen in the other urine parameters examined
Behaviour (functional findings):
effects observed, non-treatment-related
Description (incidence and severity):
There were no test substance-related or spontaneous findings in the male and female animals of test groups 1-3 during FOB observations.
There were statistically significantly increased motor activity observed in males which were without a clear relation to dosing. This effect has no toxicological relevance because the values from the substance-treated males fit to the historical control range (mean value: 245.1; range: 207.6–299.5) of male rats of this strain at a comparable age.
There were no statistically significant or clearly dose-dependent deviations concerning the overall motor activity (summation of all intervals) in the females.
A substance-induced origin for this finding is excluded with certainty because of its scattered occurrence without any relation to dosing.
Thus, the motor activity of the substance-treated males and females did not show any toxicological relevant deviations.
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Further details to histopathological findings are given in the table 7.
- liver: males of the top dose group revealed a diffuse hepatocellular hypertrophy
- reproductive organs: no histopathological findings observed
Other effects:
effects observed, non-treatment-related
Description (incidence and severity):
Clinical observations during gestation / lactation:
- discolored urine was recorded for all high and mid dose females (1000 and 300 mg/kg body weight/day) during the whole gestation and lactation period
- in 9 out of 12 high dose females salivation after treatment was noted during gestation
- in 10 out of 12 high dose females salivation after treatment was noted during lactation

No test substance-related clinical findings occurred in the female animals of test group 1
(100 mg/kg body weight/day) during the lactation period.

Both findings are not assessed as adverse or toxic effects for reasons described under overall clinical findings above.

One sperm positive control female (No. 102), one sperm positive low dose female (No. 123) and one sperm positive mid dose female (No. 136) did not deliver any F1 pups.
Reproductive function: oestrous cycle:
not examined
Reproductive function: sperm measures:
no effects observed
Description (incidence and severity):
- mean duration until sperm was detected (day 0 p.c.) amounted to 3.2 / 3.2 / 2.7 and 2.5 days (0, 100, 300 and 1000 mg/kg body weight/day)

These values reflect the normal range of biological variation inherent in the strain used in this study.
Reproductive performance:
no effects observed
Description (incidence and severity):
Male and female fertility indices are given in table 8 and 9. Mating was confirmed for all F0 males placed with females.

- not generating F1 pups: one control male (No. 2 mated with female No. 102), one low dose male (No. 23 mated with female No. 123 – 100 mg/kg body weight/day) and one mid dose male (No. 36 mated with female No. 136 – 300 mg/kg body weight/day)
- female rats not becoming pregnant (but sperm positive): control female No. 102 (mated with male No. 2), low dose female No. 123 (mated with male No. 23) and mid dose female No. 136 (mated with male No. 36)
- not pregnant females failed to show a morphological correlate for the apparent infertility for females Nos. 102 (control) and 123 (low dose)
- mid dose female No. 136 had a minimal diffuse inflammation of the uterus, which may well account for the observed infertility (see also Pathology)

The fertility indices for male and female varied between 92% (0, 100 and 300 mg/kg body weight/day) and 100% (1000 mg/kg body weight/day). These values reflect the normal range of biological variation inherent in the strain of rats used for this study. All respective values were within the range of the historical control data and do not show any relation to dosing.
Hematology determinations in high dose males revealed slightly, but statistically significantly reduced hemoglobin and hematocrit values, which are indicative of a mild anemic process. This is considered as an adverse substance-induced effect.
No treatment-related effects were noted in hematology investigations of females and serum enzyme examinations of both sexes.

The most prominent findings in clinical pathology were the reduced amounts of urine with subsequently increased specific gravity excreted by treated animals of either sex. The decreased urinary volume and the increased urinary specific gravity, however, are not considered as markers of kidney toxicity or an impairment of renal function. These findings are regarded non-renal in nature and are possibly due to decreased water intake.
A reduction in water intake could also well account for the slightly increased urea and albumin levels of the high dose males and/or females.
It is concluded that the changes in urinalysis and in blood chemistry examination are not caused by a direct toxic effect of the test compound and are not adverse in nature. This assessment is supported by the fact, that absolute and relative kidney weights as well as gross and histopathological evaluations of this organ did not give any indications for substance-induced alterations.

The livers of female animals of the top dose did not reveal any histological findings. Nevertheless, the increase in liver weight in the top dose females is thought to be substance-related.
These liver findings are regarded to be non-adverse in nature, but are considered to mirror adaptive responses to the test substance administration. Moreover, liver enzymes in these rats remained unaffected (see clinical biochemistry), but showed the usual range of biological variation.

The fertility indices observed in the study reflect the normal range of biological variation inherent in the strain of rats used for this study as all respective values were within the range of the historical control data. Furthermore, gross and histopathological examinations of these males failed to show a relevant morphological correlate for the apparently impaired fertility (see also Pathology).
Dose descriptor:
NOAEL
Remarks:
for systemic toxicity
Effect level:
300 mg/kg bw/day
Based on:
test mat.
Remarks:
equivalent to 202 mg/kg bw/day Isopropanolamine
Sex:
male
Basis for effect level:
haematology
Dose descriptor:
NOAEL
Remarks:
for systemic toxicity
Effect level:
1 000 mg/kg bw/day
Based on:
test mat.
Remarks:
equivalent to 673 mg/kg bw/day Isopropanolamine
Sex:
female
Remarks on result:
other: No effects observed for female animals.
Dose descriptor:
NOAEL
Remarks:
for reproductive performance and fertility
Effect level:
1 000 mg/kg bw/day
Based on:
test mat.
Remarks:
equivalent to 673 mg/kg bw/day Isopropanolamine
Sex:
male/female
Remarks on result:
other: no substance-related effects on fertility indices observed
Critical effects observed:
no
Clinical signs:
no effects observed
Description (incidence and severity):
- numbers of runts: 0 /0 /0 /2 in test groups 0-3 (0, 100, 300 and 1000 mg/kg body weight/day)
This finding is fully in the range of the biological variation inherent in the strain of rats used for this study.
- no adverse clinical signs up to scheduled sacrifice on day 4 post partum
Mortality / viability:
no mortality observed
Description (incidence and severity):
- viability index between days 0 - 4 p.p. varied between 98% (control) and 100% (test group 2)
- no compound related changes
Body weight and weight changes:
no effects observed
Description (incidence and severity):
- 1000 mg/kg body weight/day: mean b.w. data were statistically significantly increased on post partum day 1 (males, females and males + females) and day 4 (males and males + females)
- 100 and 300 mg/kg body weight/day: mean b.w. data were comparable to the concurrent control values
- mean pup body weight changes in all test groups: did not show any statistically significant differences
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Histopathological findings:
not examined
Other effects:
no effects observed
Description (incidence and severity):
Sex ratio:
- sex distribution and sex ratios of live F1 pups on the day of birth and 4 days post partum did not show biologically relevant differences between controls and test groups
The statistically significantly increased mean body weights at the top dose pups as well as all other differences between controls and substance-treated group in body weight data do not have any toxicological relevance and are not considered as adverse effects. Instead, they are spontaneous in nature.
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
1 000 mg/kg bw/day
Based on:
test mat.
Remarks:
equivalent to 673 mg/kg bw/day Isopropanolamine
Sex:
male/female
Remarks on result:
other: No test substance-induced signs of developmental toxicity were noted in the progeny of the F0 parents up to and including 1000 mg/kg body weight/day.
Critical effects observed:
no
Reproductive effects observed:
no

Table 5: Mean absolute weights of statistically significant changes in organs.

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

Table 6: Relative absolute weights of statistically significant changes in organs.

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)

Table 7: Histopathological findings in the liver

Liver

Male animals

Female animals

Group

0

1

2

3

0

1

2

3

Organs examined

12

12

12

12

12

 

 

12

Hypertrophy, diffuse

0

0

0

9

0

 

 

0

Table 8: Male fertility indices (F0 males) in %:

 

Test group 0

(0 mg/kg bw/day)

Test group 1

(100 mg/kg bw/day)

Test group 2

(300 mg/kg bw/day)

Test group 3

(1000 mg/kg bw/day)

concerning F1 litters

92

92

92

100

Table 9: Female fertility indices (F0 females) in %:

 

Test group 0

(0 mg/kg bw/day)

Test group 1

(100 mg/kg bw/day)

Test group 2

(300 mg/kg bw/day)

Test group 3

(1000 mg/kg bw/day)

concerning F1 litters

92

92

92

100

Conclusions:
- The NOAEL for reproductive performance and fertility is 1000 mg/kg bw/day (equivalent to 673 mg/kg bw/day Isopropanolamine) for the F0 parental rats
- The NOAEL for general, systemic toxicity is 300 mg/kg body weight/day (equivalent to 202 mg/kg bw/day Isopropanolamine) 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
Executive summary:

The hydrochloride salt of Isopropanolamine was administered orally via gavage to groups of 12 male and 12 female Wistar rats (F0 animals) at doses of 100 (673 mg/kg bw/day Isopropanolamine), 300 (202

mg/kg bw/day Isopropanolamine) and 1000 mg/kg of body weight/day (673 mg/kg bw/day Isopropanolamine) in order to detect possible effects of the test substance on the integrity and performance of the reproductive system of both sexes. Furthermore, it was intended to obtain information about the general toxicological profile including target organs and no observed adverse effect level (NOAEL) after repeated oral administration, according to the OECD 422 guideline. Control animals were dosed daily with the vehicle (tap water).

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.

F0 animals were mated 13 days after the beginning of treatment to produce a litter (F1 generation pups). Mating pairs were from the same dose group. Mating was discontinued as soon as sperm was detected in the vaginal smear, mating was discontinued. F0 animals were examined for their reproductive performance including determination of the number of implantation sites and the calculation of postimplantation loss for all F0 females.

A detailed clinical observation was performed in all animals before test substance administration and thereafter at weekly intervals. Food consumption of the F0 parents was determined regularly during premating and after the mating period and during the gestation and lactation periods in dams. In general, body weights of F0 animals were determined once a week; however, during gestation and lactation, F0 females were weighed on days 0, 7, 14 and 20 post coitum, on the parturition day and day 4 post partum. Pups were sexed on day 0 post partum and weighed one day after birth and on day 4 post partum. Their viability was recorded twice daily on each workday or only in the morning on Saturday and Sunday. All pups were sacrificed with CO2 on day 4 post partum and examined macroscopically for external and visceral findings.

Near the end of the administration period a functional observational battery was performed and motor activity was measured in 5 randomly selected parental males and females per group, respectively. Blood was sampled from 5 randomly selected parental males and 5 parental females per group for hematological and clinical chemistry examination. All surviving F0 parental animals were sacrificed by decapitation, under CO2 anesthesia, and were assessed by gross pathology. Organ weights were recorded and a histopathological examination was performed.

The following test substance-related, adverse effects/findings were noted:

- 1000 mg/kg body weight/day:

F0 parental animals: statistically significantly reduced hemoglobin and hematocrit values (i.e. indications of a mild anemic process) in the F0 males

F1 pups: no adverse effects/findings

- 100 and 300 mg/kg body weight/day:

F0 parental animals and F1 pups: no adverse effects/findings

Thus, under the conditions of this reproduction/developmental toxicity screening test the NOAEL (no observed adverse effect level) for reproductive performance and fertility is 1000 mg/kg body weight/day

(673 mg/kg bw/day Isopropanolamine) for the F0 parental rats. The NOAEL for general, systemic toxicity of the test substance is 300 mg/kg body weight/day (202 mg/kg bw/day Isopropanolamine) for the F0 parental males based on some indications for a mild anemic process.

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

No EOGRTS is available for MIPA. However, in line with Annex XI of the REACH regulation, a registrant may propose to adapt the required standard information, including a detailed justification for this adaptation. Based on the available reliable data, it is concluded in a weight-of-evidence approach, that MIPA does not induce effects on fertility (see IUCLID section 13.2: "Justification_WoE_RDT_Reprotox_Feb2019").

Screening study for reproduction/developmental toxicity (OECD 422) in rats

In a combined repeated dose oral toxicity study with a reproduction/developmental screening study (according to OECD guideline 422 and under GLP), Wistar rats (12/sex/dose) were administerd the hydrochloride salt of MIPA (CAS no. 7780 -04 -3) at 0, 100, 300 or 1000 mg/kg bw/day (equivalent to 67, 202 and 673 mg/kg bw/day MIPA) by oral gavage. 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. The fertility indices for male and female varied between 92% (0, 100 and 300 mg/kg body weight/day) and 100% (1000 mg/kg body weight/day). These values reflect the normal range of biological variation inherent in the strain of rats used for this study. All respective values were within the range of the historical control data and do not show any relation to dosing.

No adverse effects were found on reproductive and fertility parameters, thus a NOAEL of 1000 mg/kg bw/day (equivalent to 673 mg/kg bw/day MIPA, the highest dose tested) was established for effects on fertility in males and females.

Oral one generation study in rats

In a one-generation study according to FDA guidelines, Sprague-Dawley rats (25/sex/dose) were administered TIPA in the diet at 0, 500, 2000 or 7500 ppm (equivalent to a 0, 15.6, 63 or 239 mg/kg bw/d intake of MIPA in males or 0, 17.2, 71 or 275 mg/kg bw/d in females) for 5 weeks prior to mating as well as during mating, gestation and lactation. Offspring (20/sex/dose) were also administered the same doses for 90 days after weaning. Prior to weaning, these rats were exposed to TIPA in utero during pregnancy of the P rats, in maternal milk during lactation, and in any diet they consumed prior to weaning. Ophthalmological, clinical chemistry, haematology and urinalysis examinations were conducted. Histopathological examination was conducted on controls and high-dose animals for all organs and on lungs, liver, kidneys and organs with lesions at the low and intermediate doses. No adverse clinical, histological, or reproductive effects (mating index, fertility index, gestation index, gestation length, percent pups born alive, viability index, lactation index, litter survival, average number ofpups/litter, litter size) were noted.

The NOAEL was reported to be 609 mg/kg bw/d TIPA (equivalent to a 239 mg/kg bw/d intake of MIPA in males or 275 mg/kg bw/d in females) for males and 700 mg/kg bw/d TIPA for females, the highest doses tested.

Effects on developmental toxicity

Description of key information

For MIPA a PNDTS on one species (according to OECD 414) is proposed by the registrant. It is expected to further support the weight of evidence approach for toxicity to reproduction. In a combined repeated dose oral toxicity study with a reproduction/developmental screening study (performed according to OECD guideline 422) in rats exposed to the hydrochloride salt of MIPA by gavage, a NOAEL for developmental toxicity of 1000 mg/kg bw/day (equivalent to 673 mg/kg bw/day MIPA, the highest dose tested) was established. Based on the mild anemic process in males, the NOAEL for systemic toxicity is 300 mg/kg bw/day. No adverse effects were observed in the OECD414 PNDTS in rats with the structural analogues DIPA and TIPA up to the limit dose. Furthermore, developmental toxicity data for TIPA in the second species rabbit (OECD414 with Picloram TIPA salt and sub-chronic dose range finder with TIPA) also revealed no adverse developmental effects but showed, that there is a MTD (maximal tolerable dose) for the dams below the limit dose.

Link to relevant study records

Referenceopen allclose all

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
ADAPTATION ACCORDING TO REACH ANNEX XI, section 1 - see justifications attached to IUCLID section 13.2.
- Justification WoE_RDT_Reprotox_Feb2019
- Justification RA_CAS 122-20-2_Dev.tox.2ndspecies_Nov2018
Reason / purpose:
reference to same study
Reason / purpose:
reference to same study
Qualifier:
according to
Guideline:
other: OECD Test Guideline 422 - Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test
Version / remarks:
22 March 1996
Deviations:
no
GLP compliance:
yes
Limit test:
no
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and batch No.of test material: BASF and BA 1449
- Purity test date: 04 Jan 2006
- concetration of given stock solution:65.4 g isopropanolamine hydrochloride per 100 g aqueous solution

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature (freezer for reanalysis)
- Stability under test conditions: confirmed by reanalysis
- Solubility and stability of the test substance in the solvent/vehicle: stability of aq. solution confirmed by reanalysis

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: test material was weighed up and dissolved in tap water

FORM AS APPLIED IN THE TEST: solution
Species:
rat
Strain:
Wistar
Remarks:
Crl:WI(Han)
Details on test animals 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
- Diet: Ground Kliba maintenance diet mouse/rat “GLP” (Provimi Kliba SA, Kaiseraugst, Switzerland), ad libitum
- Water: ad libitum from drinking bottles
- 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:
water
Remarks:
tap water
Details on exposure:
TEST ITEM
- Dosing solution: pH 6.0 - 7.5

VEHICLE
- Amount of vehicle: 10 mL/kg bw
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
Details on mating procedure:
- Impregnation procedure: cohoused
- M/F ratio per cage: 1:1
- Length of cohabitation: maximum of 2 weeks
- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of pregnancy and the following day "day 1" post coitum
- After successful mating each pregnant female was caged: in Makrolon type M III cages (floor area about 800 cm2)from day 18 p.c. until day 4 p.p., supplied with nesting material (cellulose wadding) toward the end of pregnancy
- 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
Dose / conc.:
100 mg/kg bw/day (nominal)
Remarks:
equivalent to 67 mg/kg bw/day Isopropanolamine
Dose / conc.:
300 mg/kg bw/day (nominal)
Remarks:
equivalent to 202 mg/kg bw/day Isopropanolamine
Dose / conc.:
1 000 mg/kg bw/day (nominal)
Remarks:
equivalent to 673 mg/kg bw/day Isopropanolamine
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
Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily (littering and lactation behaviour evaluated in the mornings
- Cage side observations included any signs of morbidity, pertinent behavioral changes and signs of overt toxicity, documented for each animal

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: prior to the administration period and thereafter at weekly intervals
- Parameters: abnormal behavior during “handling”, fur, skin, salivation, nose discharge, lacrimation, pupil size, posture, palpebral closure, respiration, tremors, convulsions, abnormal movements, impairment of gait, activity/arousal level, feces (appearance/consistency), urine, other findings

BODY WEIGHT: Yes
- Time schedule for examinations:once a week at the same time of the day (in the morning);
- Time schedule exceptions: during the mating period the parental females were weighed on the day of positive evidence day 0 p.c. and on days 7, 14 and 20 p.c.; females with litter: day day 0 p.p. and on day 4 p.p.; females without a litter: weekly

FOOD CONSUMPTION AND COMPOUND INTAKE: Yes
- once a week (in a period of 7 days) for male and female parental animals
- exceptions: not determined during the mating period; F0 females with evidence of sperm were determined on days 0, 7, 14 and 20 p.c.; F0 females, which gave birth to a litter were determined on days 0 and 4 p.c.
- not determined in females without positive evidence of sperm and females without litter

POST-MORTEM EXAMINATIONS: Yes
SACRIFICE
- Male animals: after blood from 5 F0 males per group was sampled, on study day 38 necropsy of all male animals was performed
- Female animals: after blood from 5 F0 females per group was sampled, on study day 45 necropsy of all female animals was performed
GROSS NECROPSY
- Gross necropsy was not further specified.
HISTOPATHOLOGY / ORGAN WEIGHTS
- The tissues indicated in Table 1 were prepared for microscopic examination.
- The animals/organs were weighed: anesthetized animals, liver, kidneys, adrenal glands, testes, epididymides, seminal vesicle, prostate, ovaries, uterus, thymus, spleen, brain, heart

OTHER:
HAEMATOLOGY 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
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: No
- Number of implantations: Yes
- Number of early resorptions: No
- Number of late resorptions: No
Fetal examinations:
For all surviving pups (after sacrifice on day 4 post partum), all stillborn pups and those pups that died before schedule:
- External examinations: Yes: all per litter
- Soft tissue examinations: Yes: all per litter
- Skeletal examinations: No
- Head examinations: No
Statistics:
Due to limitations of characters, details for statistics are given as tables (2, 3, 4) under part 'other information'.

CLINICAL EXAMINATIONS
- DUNNETT-test (two-sided) for the hypothesis of equal means
- Pairwise comparison of each dose group with the control group using FISHER'S EXACT test for the hypothesis of equal proportions
- Pairwise comparison of each dose group with the control group using the WILCOXON-test (onesided) for the hypothesis of equal medians
- 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 (twosided) for the equal medians

CLINICAL PATHOLOGY
- 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
- Pair-wise comparison of each dose group with the control group using FISHER's exact test for the hypothesis of equal proportions

HISTOPATHOLOGY
- Means and standard deviations AND KRUSKAL-WALLIS and WILCOXON test
Indices:
FEMALE REPRODUCTION AND DELIVERY DATA
Gestation index (%) = (number of females with live pups on the day of birth / number of females pregnant*) x100
* defined as the number of females with implants in utero

Live birth index (%) = (number of liveborn pups at birth / total number of pups born) x 100

Post implantation loss (%) = ([number of implantations - number of pups delivered] / number of implantations) x 100


OFFSPRING VIABILITY INDICES
Viability index (%) = (number of live pups on day 4 after birth / number of liveborn pups on the day of birth) x 100

Sex ratio = (number of live male or female pups on day 0/4 / number of live male and female pups on day 0/4) x 100
Historical control data:
There are historical control data from various studies with the same species and strain available in-house.
The included paramters are among others: mean maternal body weights during gestation and lactation, reproduction and litter indices as indicated before, pup weights and pup necropsy data and motor activity observations.
Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
- no test substance-related or spontaneous clinical findings observed in the male and female animals of 100 mg/kg body weight/day test group
- salivation after treatment was seen in all high dose male animals (1000 mg/kg body weight/day) during study weeks 1-5, finding persisted in the respective males only for a few minutes after daily gavage dosing
- urine discoloration was observed in all male animals of the high dose group during the whole study period (week 0-5) and in all mid dose male animals (300 mg/kg body weight/day) during study weeks 1-5 and females, for all high dose females during the whole study period (week 0 - 6) and in all mid dose females during study weeks 1-6
- 7 out of 12 female animals of test group 3 showed salivation after treatment during study weeks 1-6, finding persisted in the respective females only for a few minutes after daily gavage dosing
- detailed clinical observations on study days 0, 7, 14, 21, 28, 35 and additionally day 42 for female animals did not reveal any additional, substance-induced abnormalities in the male and female animals of the test groups 0-3 (0, 100, 300 and 1000 mg/kg body weight/day)

The clinical findings, i.e. transient salivation and discolored urine are considered to be substance-induced, but are without any toxicological relevance and are not assessed as being adverse.
It is assumed that the temporary salivation was induced by the bad taste of the test substance (small droplets of the test substance solution might be adjacent on the tip of the gavage) or minor local affection of the upper digestive tract (without showing a morphological correlate at pathology). The urine discoloration is possibly related to a chemical reaction of the test substance or its metabolites with the bedding or with components of the air.
Mortality:
no mortality observed
Description (incidence):
There were no substance-related or spontaneous mortalities in any of the groups.
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
Body weights and body weight gains of all male and female (+ during the gestation, lactation period and after weaning) animals of all substance-treated groups (100, 300 and 1000 mg/kg body weight/day) were comparable to that of the concurrent control group during the whole study taking normal biological variability into account.
The statistically significantly increased body weight gains of the low and mid dose females (100 and 300 mg/kg body weight/day) during premating weeks 1-2 are considered to be spontaneous in nature and not to be adverse.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
There were no effects observed for food consumption compared to the control group.
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
- slightly, but statistically significantly decreased hemoglobin and hematocrit values in the peripheral blood of high dose males
- no treatment related effects were seen in the other hematology parameters of males and in the hematology investigations of females
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
- differences in serum enzyme activities were not evident at any dose level in either males or females
- blood chemistry results showed statistically significantly increased urea concentrations in the serum of high dose males and significantly higher albumin levels in the serum of high dose females
- marginally increased urea concentrations were also seen in the high dose females
- the increase in females was not sufficient to be statistically identified
- no treatment related changes were found in the other blood chemistry parameters
Urinalysis findings:
effects observed, non-treatment-related
Description (incidence and severity):
- decreased amounts of urine with increased specific gravity in all animals, more pronounced in females than in males
- no treatment related effects were seen in the other urine parameters examined
Behaviour (functional findings):
effects observed, non-treatment-related
Description (incidence and severity):
There were no test substance-related or spontaneous findings in the male and female animals of test groups 1-3 during FOB observations.
There were statistically significantly increased motor activity observed in males which were without a clear relation to dosing. This effect has no toxicological relevance because the values from the substance-treated males fit to the historical control range (mean value: 245.1; range: 207.6–299.5) of male rats of this strain at a comparable age.
There were no statistically significant or clearly dose-dependent deviations concerning the overall motor activity (summation of all intervals) in the females.
A substance-induced origin for this finding is excluded with certainty because of its scattered occurrence without any relation to dosing.
Thus, the motor activity of the substance-treated males and females did not show any toxicological relevant deviations.
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Description (incidence and severity):
Detailed information on changes (absolute and relative) of organ weights are given in the table 5 and 6 under any other information.

ABSOLUTE WEIGHT EFFECTS
- statistically increased brain weight of the mid dose group of female animals
This effect is thought to be incidental in nature, as no relevant microscopic findings were noted and no dose response relationship was observed.
- statistically increased thymus weights of females of the mid and top dose group
This effect is also thought to be incidental as no microscopic findings were noted.

RELATIVE WEIGHT EFFECTS
- statistically significant increased brain and kidney weights of females of the mid dose group
This effect was considered incidental in nature, as there was no dose-response relationship and no relevant microscopic findings were noted.
- statistically increased thymus weights of females of the mid and top dose group
This effect is also thought to be incidental as no microscopic findings were noted.
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
- additional statistically significant intergroup differences in the results of clinical pathology testing
- these deviations were marginal, incidental or inconsistent, when compared with the other sex, and/or lack dose-response relationship

These findings were considered to be of no toxicological significance.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Further details to histopathological findings are given in the table 7.
- liver: males of the top dose group revealed a diffuse hepatocellular hypertrophy
- reproductive organs: no histopathological findings observed
Other effects:
effects observed, non-treatment-related
Description (incidence and severity):
Clinical observations during gestation / lactation:
- discolored urine was recorded for all high and mid dose females (1000 and 300 mg/kg body weight/day) during the whole gestation and lactation period
- in 9 out of 12 high dose females salivation after treatment was noted during gestation
- in 10 out of 12 high dose females salivation after treatment was noted during lactation

No test substance-related clinical findings occurred in the female animals of test group 1 (100 mg/kg body weight/day) during the lactation period.

Both findings are not assessed as adverse or toxic effects for reasons described under overall clinical findings above.
Number of abortions:
not specified
Pre- and post-implantation loss:
effects observed, non-treatment-related
Description (incidence and severity):
- postimplantation loss in the high dose group slightly exceeded the historical control range, however, the mean number of F1 pups delivered per dam remained unaffected (12.9 / 11.6 / 10.7 and 11.5 pups/dam at 0, 100, 300 and 1000 mg/kg body weight/day)
- no statistically significant differences between the groups for the postimplantation loss
Dead fetuses:
no effects observed
Description (incidence and severity):
- rate of liveborn pups: live birth indices ranging between 99% (high and low dose) and 100% (control and mid dose)
- rate of stillborn pups was comparable between the groups
Changes in number of pregnant:
no effects observed
Description (incidence and severity):
- gestation index reached 100% in all test groups
Other effects:
no effects observed
Description (incidence and severity):
- mean number of implantation sites was comparable between all test groups including the controls and did not show any relation to dosing (13.6 / 12.4 / 11.8 and 13.2 implants/dam in test groups 0 - 3 (0, 100, 300 and 1000 mg/kg body weight/day))
Details on maternal toxic effects:
Hematology determinations in high dose males revealed slightly, but statistically significantly reduced hemoglobin and hematocrit values, which are indicative of a mild anemic process. This is considered as an adverse substance-induced effect.
No treatment-related effects were noted in hematology investigations of females and serum enzyme examinations of both sexes.
The most prominent findings in clinical pathology were the reduced amounts of urine with subsequently increased specific gravity excreted by treated animals of either sex. The decreased urinary volume and the increased urinary specific gravity, however, are not considered as markers of kidney toxicity or an impairment of renal function. These findings are regarded non-renal in nature and are possibly due to decreased water intake.
A reduction in water intake could also well account for the slightly increased urea and albumin levels of the high dose males and/or females.
It is concluded that the changes in urinalysis and in blood chemistry examination are not caused by a direct toxic effect of the test compound and are not adverse in nature. This assessment is supported by the fact, that absolute and relative kidney weights as well as gross and histopathological evaluations of this organ did not give any indications for substance-induced alterations.
The livers of female animals of the top dose did not reveal any histological findings. Nevertheless, the increase in liver weight in the top dose females is thought to be substance-related.
These liver findings are regarded to be non-adverse in nature, but are considered to mirror adaptive responses to the test substance administration. Moreover, liver enzymes in these rats remained unaffected (see clinical biochemistry), but showed the usual range of biological variation.

Considering the unaffected implantation and litter size and the absence of any statistically significant differences between the groups for the postimplantation loss, no test substance-induced intrauterine embryo-/fetolethality is assumed. The rate of liveborn pups was also not affected by the test substance administration as indicated by live birth indices ranging between 99% (high and low dose) and 100% (control and mid dose). Moreover, the rate of stillborn pups was comparable between the groups.
Thus, the administration of the Hydrochloride salt of Isopropanolamine did not adversely affect reproduction and delivery of the F0 females.
Dose descriptor:
NOAEL
Remarks:
for systemic toxicity
Effect level:
300 mg/kg bw/day (nominal)
Based on:
test mat.
Remarks:
equivalent to 202 mg/kg bw/day Isopropanolamine
Basis for effect level:
haematology
Dose descriptor:
NOAEL
Remarks:
for systemic toxicity
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Remarks:
equivalent to 673 mg/kg bw/day Isopropanolamine
Remarks on result:
other: No effects observed for female animals.
Dose descriptor:
NOAEL
Remarks:
for developmental toxicity
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Remarks:
equivalent to 673 mg/kg bw/day Isopropanolamine
Remarks on result:
other: No effects observed for maternal developmental toxicity
Abnormalities:
no effects observed
Fetal body weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
- mean pup body weights in test group 3 (1000 mg/kg body weight/day) were statistically significantly increased on post partum day 1 (males, females and males + females) and day 4 (males and males + females)
- mean pup body weights in test groups 1 and 2; 100 and 300 mg/kg body weight/day were comparable to the concurrent control values
- mean pup body weight changes in test groups 1, 2 and 3; 100, 300 and 1000 mg/kg body weight/day did not show any statistically significant differences

- body weight and body weight changes: spontaneous in nature, are not of toxicological relevance and are not considered as adverse effects
Reduction in number of live offspring:
no effects observed
Description (incidence and severity):
- mean number of delivered pups per dam and the rate of liveborn and stillborn pups were evenly distributed among the groups
Changes in sex ratio:
no effects observed
Description (incidence and severity):
- sex distribution and sex ratios of live F1 pups on the day of birth and 4 days post partum did not show biologically relevant differences between controls and test groups
Changes in litter size and weights:
no effects observed
Description (incidence and severity):
- unaffected litter size between the groups
Changes in postnatal survival:
no effects observed
Description (incidence and severity):
- viability index as indicator for pup mortality between days 0 - 4 p.p. varied between 98% (control) and 100% (test group 2)
- no compound related changes
External malformations:
no effects observed
Description (incidence and severity):
- numbers of runts were 0 /0 /0 /2 in test groups 0-3 (0, 100, 300 and 1000 mg/kg body weight/day), fully in the range of the biological variation inherent in the strain of rats
- scattered occurrence without a clear relation to dosing of post mortem autolysis
Visceral malformations:
effects observed, non-treatment-related
Description (incidence and severity):
- very few findings as situs inversus, misshapen spleen, infarct of liver, empty stomach, and dilated renal pelvis were of scattered occurrence without a clear relation to dosing
- substance-induced origin for these findings is excluded due to existence in the historical control data at comparable or even higher incidences
Details on embryotoxic / teratogenic effects:
No test substance-induced signs of developmental toxicity were noted in the progeny of the F0 parents up to and including 1000 mg/kg body weight/day. The number of delivered F1 pups/litter, their postnatal survival and their body weights remained unaffected by the test substance. Clinical and/or gross necropsy examinations of the F1 pups revealed only findings, which were considered to be spontaneous in nature.
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Remarks:
equivalent to 673 mg/kg bw/day Isopropanolamine
Sex:
male/female
Remarks on result:
other: No effects observed for F1 animals.
Abnormalities:
no effects observed
Developmental effects observed:
no

- 1000 mg/kg body weight/day, F1 pups: no adverse effects/findings

- 100 and 300 mg/kg body weight/day, F1 pups: no adverse effects/findings

Conclusions:
- Only the findings of decreased hemoglobin and hematocrit at 1000 mg/kg bw/day (equivalent to 673 mg/kg bw/day Isopropanolamine) 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.
Executive summary:

The hydrochloride salt of Isopropanolamine was administered orally via gavage to groups of 12 male and 12 female Wistar rats (F0 animals) at doses of 100 (673 mg/kg bw/day Isopropanolamine), 300 (202

mg/kg bw/day Isopropanolamine) and 1000 mg/kg of body weight/day (673 mg/kg bw/day Isopropanolamine) in order to detect possible effects of the test substance on the integrity and performance of the reproductive system of both sexes. Furthermore, it was intended to obtain information about the general toxicological profile including target organs and no observed adverse effect level (NOAEL) after repeated oral administration, according to the OECD 422 guideline. Control animals were dosed daily with the vehicle (tap water).

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.

F0 animals were mated 13 days after the beginning of treatment to produce a litter (F1 generation pups). Mating pairs were from the same dose group. Mating was discontinued as soon as sperm was detected in the vaginal smear, mating was discontinued. F0 animals were examined for their reproductive performance including determination of the number of implantation sites and the calculation of postimplantation loss for all F0 females.

A detailed clinical observation was performed in all animals before test substance administration and thereafter at weekly intervals. Food consumption of the F0 parents was determined regularly during premating and after the mating period and during the gestation and lactation periods in dams. In general, body weights of F0 animals were determined once a week; however, during gestation and lactation, F0 females were weighed on days 0, 7, 14 and 20 post coitum, on the parturition day and day 4 post partum. Pups were sexed on day 0 post partum and weighed one day after birth and on day 4 post partum. Their viabilitys recorded twice daily on each workday or only in the morning on Saturday and Sunday. All pups were sacrificed with CO2 on day 4 post partum and examined macroscopically for external and visceral findings.

Near the end of the administration period a functional observational battery was performed and motor activity was measured in 5 randomly selected parental males and females per group, respectively. Blood was sampled from 5 randomly selected parental males and 5 parental females per group for hematological and clinical chemistry examination. All surviving F0 parental animals were sacrificed by decapitation, under CO2 anesthesia, and were assessed by gross pathology. Organ weights were recorded and a histopathological examination was performed.

The following test substance-related, adverse effects/findings were noted:

- 1000 mg/kg body weight/day:

F0 parental animals: statistically significantly reduced hemoglobin and hematocrit values (i.e. indications of a mild anemic process) in the F0 males

F1 pups: no adverse effects/findings

- 100 and 300 mg/kg body weight/day:

F0 parental animals and F1 pups: no adverse effects/findings

Thus, under the conditions of this reproduction/developmental toxicity screening test the NOAEL (no observed adverse effect level) for reproductive performance and fertility is 1000 mg/kg body weight/day

(673 mg/kg bw/day Isopropanolamine) for the F0 parental rats. The NOAEL for general, systemic toxicity of the test substance is 300 mg/kg body weight/day (202 mg/kg bw/day Isopropanolamine) for the F0 parental males based on some indications for a mild anemic process.

The NOAEL for parental females (F0) was found to be 1000 mg/kg body weight/day (673 mg/kg bw/day Isopropanolamine).

The NOAEL for developmental toxicity was 1000 mg/kg body weight/day (673 mg/kg bw/day Isopropanolamine).

Endpoint:
developmental toxicity
Type of information:
experimental study planned
Justification for type of information:
TESTING PROPOSAL ON VERTEBRATE ANIMALS
The proposed testing for a pre-natal developmental toxicity study (PNDTS) in rats is a standard information requirement according to Annex IX of the REACH regulation 1907/2006. One species (rat) in the most appropriate route (oral) of administration in regard to the likely route of human exposure according to OECD 414 is proposed. Based on the results of this study, which are expected to support the weight of evidence approach for RDT and toxicity to reproduction presented in the dossier, further testing in a second species might not be required, as this standard information requirement could be covered by the weight of evidence approach.

NON-CONFIDENTIAL NAME OF SUBSTANCE:
- Name of the substance on which testing is proposed to be carried out : 1-aminopropan-2-ol (Monoisopropanolamin: MIPA)

CONSIDERATIONS THAT THE GENERAL ADAPTATION POSSIBILITIES OF ANNEX XI OF THE REACH REGULATION ARE NOT ADEQUATE TO GENERATE THE NECESSARY INFORMATION:
- Available GLP studies: There is no GLP PNDTS available for MIPA.
- Available non-GLP studies : There is no non-GLP PNDTS available for MIPA.
- Historical human data : There are no human data available covering the endpoint of pre-natal developmental toxicity.
- (Q)SAR: There are no valid qualitative or quantitative structure-activity relationship models available that are scitifically accepted across a broad majority for this endpoint.
- In vitro methods : There are no sufficiently well developed in vitro methos existing according to internationally agreed test development criteria.
- Weight of evidence/read-across : There are structurally similar substances existing with properties (physicochemical and toxicological), likely to be similar to MIPA. Based on these information the standard requirement of Annex X (PNDTS, second species) may be predicted. However, as no data for the potential effects on pre-natal development exists, testing in one species is considered to be necessary to minimize the remaining uncertainty for human health.

CONSIDERATIONS THAT THE SPECIFIC ADAPTATION POSSIBILITIES OF ANNEXES VI TO X (AND COLUMN 2 THEREOF) OF THE REACH REGULATION ARE NOT ADEQUATE TO GENERATE THE NECESSARY INFORMATION:
- The substance is not known to be a genotoxic carcinogen, to be a germ cell mutagen but evidence exists that the substance is of toxicity seen in experimental tests.
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Species:
rat
Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Acceptable, well-documented publication which meets basic scientific prnciples.
Justification for type of information:
ADAPTATION ACCORDING TO REACH ANNEX XI, section 1 - see justifications attached to IUCLID section 13.2.
- Justification WoE_RDT_Reprotox_Feb2019
- Justification RA_CAS 122-20-2_Dev.tox.2ndspecies_Nov2018
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Principles of method if other than guideline:
Groups of 25 time-mated female Sprague–Dawley rats were given 0, 100, 300, or 1000 mg DIPA/kg/day on gestation days 6 through 20 by oral gavage.
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
other: CRL:CD(SD)
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories Inc. (Kingston, New York and Raleigh, North Carolina)
- Diet: LabDiet #5002 Certified Rodent Diet (PMI Nutrition International, St. Louis, MO) ad libitum
- Water: tap water ad libitum



ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21-25 °C
- Humidity (%): 45-60 %
- Air changes (per hr): 12-15
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
water
Analytical verification of doses or concentrations:
yes
Details on mating procedure:
- Impregnation procedure: purchased timed pregnant
Duration of treatment / exposure:
Gestation days 6-20
Frequency of treatment:
daily
Duration of test:
21 days
Remarks:
Doses / Concentrations:
0, 100, 300 or 1000 mg/kg bw/day
Basis:
nominal conc.
No. of animals per sex per dose:
25 (females only)
Control animals:
yes, concurrent vehicle
Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily

BODY WEIGHT: Yes

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day # 21
- Organs examined: The weights of the kidneys, liver and gravid uteri were obtained and the kidneys, liver and all gross lesions were preserved in 10% NBF.
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of resorptions: Yes

Fetal examinations:
- External examinations: Yes: all per litter
- Soft tissue examinations: Yes: half per litter
- Skeletal examinations: Yes: half per litter
- Head examinations: Yes: half per litter
Statistics:
Means and standard deviations were calculated for all continuous data. These parameters were first examined for equality of variance using Bartlett’s test (a = 0.01; Winer, 1971). If the results of the Bartlett’s test were significant, the data were transformed in an attempt to obtain equality of variances. The order of transformations used was the common log, the inverse and the square root with the best fit used for subsequent statistical testing. Maternal body weight, weight gain, feed consumption, organ weights and fetal body weights were analyzed using parametric or non-parametric ANOVA followed by Dunnett's or Wilcoxon's test for comparison to controls; pre- and post-implantation loss were evaluated using a Censored Wilcoxon's test; Corpora lutea, implantations and litter size were analyzed using non-parametric ANOVA follewed by Wilcoxon's test; pregnancy rates were evaluated using Fisher exact probability test; and fetal sex ratios were analyzed using a binomial distribution test. As statistical interactions (i.e., time by dose or sex by dose) were identified in several of the tests, the alpha levels for interaction terms were set a priori at 0.01–0.10, with Bonferroni’s correction. The alpha level for comparison of individual dose groups to controls was set a priori at 0.05.
Details on maternal toxic effects:
Maternal toxic effects:no effects
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Basis for effect level:
other: maternal toxicity
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Basis for effect level:
other: fetotoxicity
Abnormalities:
not specified
Developmental effects observed:
not specified
Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
27 Jan 1994 - 21 Feb 1994
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
ADAPTATION ACCORDING TO REACH ANNEX XI, section 1 - see justifications attached to IUCLID section 13.2.
- Justification WoE_RDT_Reprotox_Feb2019
- Justification RA_CAS 122-20-2_Dev.tox.2ndspecies_Nov2018
Qualifier:
according to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
GLP compliance:
yes
Limit test:
no
Specific details on test material used for the study:
- Name of test material (as cited in study report): Triisopropanolamine
- Analytical purity: 92 %
- Lot/batch No.: 10-4852
- Storage condition of test material: Room temperature, under nitrogen atmosphere
Species:
rat
Strain:
Wistar
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: THOMAE, Biberach an der Riss, FRG
- Age at study initiation: 77-89 days
- Weight at study initiation: 242 g (mean)
- Housing: singly in type DK III stainless steel wire mesh cage.
- Diet: ground Kliba 343 feed, Klingenthalmuehle AG, Kaiseraugst, Switzerland, ad libitum
- Water: tap water, ad libitum
- Acclimation period: at least 5 days

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

Route of administration:
oral: gavage
Vehicle:
water
Remarks:
double distilled
Details on exposure:
Doses of 0, 100, 400 and 1000 mg/kg bw/day were administered in a volume of 10 mL/kg, at a concentration of 0, 1000, 4000 and 10000 mg/100mL, respectively.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Test substance: purity and stability were analyzed by gas chromatography (GC); and homogeneity was proven visually.
Test solutions: analysis of stability (for 3 hrs) in double distilled water was carried out in a range-finding study. Concentrations were analyzed twice during the study by GC. Test solutions were freshly prepared on the day of dosing.
Details on mating procedure:
- Impregnation procedure: cohoused
- If cohoused:
- M/F ratio per cage: 1/1
- Length of cohabitation: overnight
- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of pregnancy
Duration of treatment / exposure:
on day 6 through day 15 post coitum (p.c.)
(Sacrifice on day 20 p.c.)
Frequency of treatment:
once a day during the period of major organogenesis (day 6 to day 15 p.c.)
Duration of test:
20 days
Dose / conc.:
100 mg/kg bw/day (nominal)
Dose / conc.:
400 mg/kg bw/day (nominal)
Dose / conc.:
1 000 mg/kg bw/day (nominal)
No. of animals per sex per dose:
25
Control animals:
yes, concurrent vehicle
Maternal examinations:
CAGE SIDE OBSERVATIONS
- Time schedule: once a day and more often when clinical signs of toxicity were elicited. Mortality was check twice a day on workdays and once a day during weekends and public holidays.

BODY WEIGHT and FOOD CONSUMPTION
- Time schedule for examinations: days 0 (only bw), 1, 3, 6, 8, 10, 13, 15, 17, 20 p.c.

POST-MORTEM EXAMINATIONS
- Sacrifice on gestation day # 20
- Organs examined: uterus and ovaries after gross pathology

OTHER: The correct body weight gain was calculated after terminal sacrifice.

Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
- Other: furthermore, calculations of conception rate and pre- and postimplantation losses were carried out
Fetal examinations:
- External examinations: Yes: all per litter: fetus was weighed, sexed, examined macroscopically for external findings, and viability, condition of the placentae, umbilical cords, fetal membranes and fluids were examined.
- Soft tissue examinations: Yes: half per litter
- Skeletal examinations: Yes: half per litter
- Head examinations: No
Statistics:
The data were evaluated statistically using the computer systems of the Department of Toxicology of BASF AG. The Dunnett-test was used for a simultaneous comparison of several dose groups with the control. The hypothesis of equal means was tested. This test was performed two-sided and was used for the statistical evaluation of the following parameters: food consumption, body weight, body weight change, corrected body weight gain (net maternal body weight change), weight of the uterus before it was opened, number of corpora lutea, number of implantations, number of resorptions and number of live fetuses, proportion of preimplantation loss, postimplantation loss, resorptions and live fetuses in each litter, litter mean fetal body weight and litter mean placental weight. Fisher's Exact Test was used for a pairwise comparison of each dose group with the control for the hypothesis of equal proportions. This test was performed one-sided and was used for female mortality, females pregnant at terminal sacrifice and the number of litters with fetal findings. The Wilcoxon-Test was used for a comparison of each dose group with the control for the hypothesis of equal medians. This test was performed one-sided and was used for the proportion of fetuses with malformations, variations, retardations and/or unclassified observations in each litter. If the results of these tests were significant, labels (* for p< 0.05, ** for p< 0.01) were printed in the summary tables.
Clinical signs:
no effects observed
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
There were no statistically significant differences between the controls and the substance-treated dams concerning mean body weights. At the beginning of the treatment period (days 6-8 p.c.), however, the dams of the highest dose group (1,000 mg/kg bw/day) showed a statistically significantly reduced body weight gain (only about 36% of the weight gain of the concurrent control group; see table in remarks on results). The corrected body weight gain (terminal body weight on day 20 p.c. minus weight of the uterus before it was opened minus body weight on day 6 p.c.) was statistically significantly lower in the high dose group (about 87% of the value of the concurrent control group), which is related to the test substance administration (see table in remarks on results).
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
In the 1000 mg/kg group statistically significantly decreased food consumption at the beginning of the treatment period (days 6-10 p.c.; about 13% (days 6 to 8 p.c.) or about 16% (days 8 to 10 p.c.) lower than the values of the concurrent control group.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
not examined
Other effects:
not examined
Number of abortions:
no effects observed
Pre- and post-implantation loss:
no effects observed
Total litter losses by resorption:
no effects observed
Early or late resorptions:
no effects observed
Dead fetuses:
no effects observed
Changes in pregnancy duration:
not examined
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): no effects observed
Changes in number of pregnant:
no effects observed
Other effects:
not examined
Details on maternal toxic effects:
There were no substance-related and/or biologically relevant differences between the groups in conception rate, in the mean number of corpora lutea and implantation sites or in the values calculated for the pre- and the postimplantation losses, the number of resorptions and viable fetuses. The differences evident are considered to be incidental and within the normal range of deviations for animals of this strain and age.
Key result
Dose descriptor:
NOAEL
Effect level:
400 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
body weight and weight gain
food consumption and compound intake
Key result
Abnormalities:
no effects observed
Fetal body weight changes:
no effects observed
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): no effects observed
Reduction in number of live offspring:
no effects observed
Changes in sex ratio:
no effects observed
Changes in litter size and weights:
no effects observed
Changes in postnatal survival:
no effects observed
External malformations:
no effects observed
Skeletal malformations:
effects observed, non-treatment-related
Description (incidence and severity):
The few statistically significant differences on embryo-/fetotoxicity, which occurred were exclusively related to fetal skeletal variations and retardations. These findings consisted of: an increased rate of affected fetuses/litter and an increased litter incidence with shortened 13th rib(s) at 100, 400 and 1,000 mg/kg bw/day, respectively; a consequently increased rate of low and intermediate dose fetuses/litter with total skeletal variations; an increased litter incidence of incompletely ossified or smaller sternebra(e) in the 400 mg/kg body weight group. These findings are considered to be spontaneous in nature because no dose-response relationship is given and/or the respective values are fully in the historical control range.
Visceral malformations:
no effects observed
Other effects:
not examined
Key result
Remarks on result:
not determinable due to absence of adverse toxic effects
Key result
Abnormalities:
no effects observed
Key result
Developmental effects observed:
no

Mean maternal body weight change during gestation (grams):

   0 mg/kg bw/d(n = 25) 100 mg/kg bw/d(n = 24) 400 mg/kg bw/d(n = 25) 1000 mg/kg bw/d(n = 23)   
Days 0 - 1  4.3±3.76 3.7±3.60  4.3±3.41  4.3±2.89   
Days 1 - 3   11.7±3.56  11.9±3.41  11.7±3.77  11.8±2.95  
Days 3 - 6   13.0±4.56  12.1±4.40  13.1±4.37  12.5±3.39  
Days 6 - 8  7.5±3.90  7.4±4.21  5.0±4.26  2.7±3.93**  
Days 8 - 10  9.9±4.59  9.4±5.15  11.4±4.43 8.0±3.72   
Days 10 - 13   17.5±3.57  17.1±4.92  19.1±5.42  19.0±4.98  
Days 13 - 15  11.7±4.51  11.9±4.15  12.5±4.36  13.7±4.62  
Days 15 - 17  25.4±6.47  25.8±5.23  25.3±4.38  26.6±3.60  
Days 17 - 20  50.3±10.57  49.2±6.64 51.8±7.58   52.6±5.56  

**: p 0.01; Dunnett-test.

Mean gravid uterine Weights and net maternal body weight change (grams):

0 mg/kg bw/d(n = 25)   100 mg/kg bw/d(n = 24) 400 mg/kg bw/d(n = 25) 1000 mg/kg bw/d(n = 23)   
Gravid uterus  78.3±19.22  80.3±10.49 82.5±13.18   84.4±10.16  
Carcass  317.7±22.09  310.8±20.66 314.3±22.22   306.2±13.17  
Net weight change from day 6  44.1±8.54  40.4±7.52  42.6±7.06  38.2±6.72*  

*: p 0.05; Dunnett-test.

Under the conditions of this full-scale prenatal toxicity study, the administration of Triisopropanolamine to pregnant female Wistar rats during organogenesis elicited overt signs of maternal toxicity at 1000 mg/kg body weight/day, while 100 or 400 mg/kg body weight/day were tolerated by the dams without any substance-induced findings. The gestational parameters were not influenced by the test substance administration in any of the three test groups.

No signs of embryo-/fetotoxicity, especially no substance-induced indications of teratogenicity were noted up to and including the dose of 1,000 mg/kg body weight/day.

Therefore, the no observed adverse effect level (NOAEL) for the dams in this full-scale prenatal toxicity study is 400 mg/kg body weight/day, while it is 1000 mg/kg body weight/day for the fetal organism.

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
ADAPTATION ACCORDING TO REACH ANNEX XI, section 1 - see justifications attached to IUCLID section 13.2.
- Justification WoE_RDT_Reprotox_Feb2019
- Justification RA_CAS 122-20-2_Dev.tox.2ndspecies_Nov2018
Qualifier:
according to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Version / remarks:
1981
Qualifier:
according to
Guideline:
EPA OPP 83-3 (Prenatal Developmental Toxicity Study)
GLP compliance:
yes
Limit test:
no
Specific details on test material used for the study:
- Test material name: 4-Amino-3.5,6-trichloropicolinate, Triisopropanolamine Salt
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Hazleton Research Products, Inc. (Kalamazoo, MI)
- Age at study initiation: 6 - 8 weeks
- Weight at study initiation: 3250 – 4250 g
- Housing: individually in cages with wire floors
- Diet: 8 oz/day, Certified Laboratory Rabbit Chow No. 5322 (Purina Mills, Inc., St. Louis, MO)
- Water: ad libitum, municipal tap water
- Acclimation period: at least 2 weeks
Route of administration:
oral: gavage
Vehicle:
water
Remarks:
deionized
Details on exposure:
VEHICLE
- Amount of vehicle (if gavage): 2 mL/kg bw
Analytical verification of doses or concentrations:
yes
Details on mating procedure:
- Impregnation procedure: artificial insemination
Duration of treatment / exposure:
Days 7 - 19 of gestation
Frequency of treatment:
daily
Duration of test:
Until day 28 of gestation
Dose / conc.:
54 mg/kg bw/day (nominal)
Remarks:
Phase II
Dose / conc.:
180 mg/kg bw/day (nominal)
Remarks:
Phase I & II
Dose / conc.:
538 mg/kg bw/day (nominal)
Remarks:
Phase I & II
Dose / conc.:
1 000 mg/kg bw/day (nominal)
Remarks:
Phase I & II
No. of animals per sex per dose:
18
Control animals:
yes, concurrent vehicle
Maternal examinations:
CAGE SIDE OBSERVATIONS:
Animals were observed daily. Any animal which showed indications of early termination of pregnancy was submitted for a complete necropsy.

BODY WEIGHT:
Body weights were recorded on Days 0, daily during the dosing period, and on Days 20 and 28 of gestation

FOOD CONSUMPTION: only in Phase II study
Feed consumption data was collected daily throughout the test period to better interpret the results obtained from body weight, body weight gain and fecal output data.

POST-MORTEM EXAMINATIONS:
- Sacrifice on gestation day # 28
- Examinations: a limited gross pathologic examination (necropsy) was performed on all animals and liver with gallbladder, kidneys and gravid uterine weights were recorded. Any obvious structural or gross pathologic alterations noted in the adult were recorded. Sections of liver with gallbladder, kidneys and gross lesions were preserved in neutral, phosphate-buffered 10% formalin. Since there were no gross pathologic observations considered. treatment-related. in the dams at any dose level, tissues were not examined histopathologically.
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
- Other: position of fetuses in utero and position of resorption sites
Fetal examinations:
- External examinations: Yes: The sex and body weight of each fetus was determined. Any gross external alterations were recorded.
- Soft tissue examinations: Yes: All fetuses were examined by dissection under a low power stereomicroscope for evidence of visceral alterations. This examination also included a fresh examination of the brain.
- Skeletal examinations: Yes: All fetuses were then preserved in alcohol, eviscerated, cleared and stained with alizarin red-S and examined for skeletal alterations.
- Head examinations: Yes
Statistics:
Descriptive statistics (means and standard deviations) were calculated for feed consumption. Maternal body weights, body weight gains, organ weights (absolute and relative) and fetal body weights were evaluated by Bartlett's test for equality of variances. Based on the outcome of Bartlett's test, a parametric or nonparametric analysis of variance (ANOVA) was performed. If the ANOVA was significant, analysis by Dunnett's test or the Wilcoxon Rank-Sum test with Bonferroni's correction was performed, respectively. Statistical evaluation of the frequency of pre-implantation loss, resorptions and fetal alterations among litters and the fetal population was performed using a censored Wilcoxon test with Bonferroni's correction. The number of corpora lutea and implants, and litter size were evaluated using a nonparametric ANOVA followed by the Wilcoxon Rank-Sum test with Bonferroni's correction. Pregnancy rates were analyzed using the Fisher exact probability test. Fetal sex ratios were analyzed using a binomial distribution test. Statistical outliers were identified using a sequential method, but were not excluded unless justified by sound scientific reasons unrelated to treatment.
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
- Phase I: The only treatment-related in-life alteration observed was an increase in the incidence of decreased feces in rabbits given 538 and 1000 mg/kg bw/day. This alteration, though transient, lasting 1-4 days, was considered to be a treatment-related response since it was accompanied by a corresponding decrease in body weight gain.
- Phase II: Dose-related increases in the incidence of decreased feces were observed in treated rabbits during the treatment period. Soft feces was also common among the treated rabbits. Perineal soiling was seen in 7 out of 18 high-dose rabbits.
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
- Phase I: There were no significant treatment-related effects on body weights. Dose-related statistically significant decreases in body weight gain were observed in dams given 180, 538 and 1000 mg/kg bw/day during Days 7 through 10 of gestation. The body weight gain of dams given 1000 mg/kg bw/day was also statistically decreased during Days 7-20. A compensatory increase in body weight gain following the exposure period (Days 20 through 28) was observed in dams given 180, 538 and 1000 mg/kg bw/day. These compensatory increases in body weight gain were statistically significant in dams given 538 and 1000 mg/kg bw/day and resulted in equal weight gains for all groups of rabbits over the test period (Days 0 through 28).
- Phase II: Body weights of the high-dose dams were decreased throughout most of the dosing period, with statistically significant decreases occurring on Days 10, 16 and 20. The decreased weights were consistent with and reflective of the decreased feed consumption noted in these animals during this time period. No significant effect on body weights was observed in dams given 54, 180 or 538 mg/kg bw/day. Dose-related statistically significant decreases in body weight gains were observed in dams given 180,538 and 1000 mg/kg bw/day at the start of the dosing period (Days 7-10). The decrease in body weight gain in the 180 mg/kg/day dose group dams was not considered to be of biological significance since body weight gain after Day 10 of gestation increased and was higher than the controls for the rest of the test period (Days 10-28). The 538 and 1000 mg/kg bw/day dose group dams had, in general, lower body weight gain during the later part of the dosing period, resulting in a statistically decreased body weight gain during Days 7-20. A statistically significant compensatory increase in body weight gain was seen in the 538 and 1000 mg/kg/ day dose group rabbits following the exposure period (Days 20-28). The changes in body weight gain paralleled the trends in feed consumption.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
- Phase I: Although feed consumption was not recorded in Phase I of the study, the decreased body weight gain in all treated rabbits during Days 7 through 10 and decreased fecal output at the 538 and 1000 mg/kg bw/day dose levels indicate a reduction in feed consumption at the start of the dosing period in these rabbits.
- Phase II: There was a dose-related decrease in feed consumption in dams given 538 and 1000 mg/kg bw/day during the dosing period. Feed consumption was also slightly decreased during Days 7-10 in dams given 180 mg/kg bw/day. No significant effect on feed consumption was observed in dams given 54 mg/kg/day. Feed consumption of all treated groups of rabbits was comparable to the control during the post-dosing period.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
- Phase I & II: There were no significant treatment-related effects on absolute or relative liver or kidney weights at any dose level.
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
- Phase I: Gross pathologic examination revealed no remarkable alterations other than a pale liver, decreased ingesta and blood in the urine.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Other effects:
not examined
Number of abortions:
effects observed, treatment-related
Description (incidence and severity):
- Phase II: Two rabbits from the 1000 mg/kg bw/day dose group aborted their fetuses and were sent for a complete necropsy.
Pre- and post-implantation loss:
no effects observed
Total litter losses by resorption:
no effects observed
Early or late resorptions:
effects observed, treatment-related
Description (incidence and severity):
- Phase I: The resorption rates of dams given 180, 538 or 1000 mg/kg/day were increased relative to the control, with statistically significant increases in dams given 180 or 1000 mg/kg/day. The apparent increases in resorption rates of dams given the test substance were not considered to be treatment-related since, 1) the resorption rates for all dose groups were within the range of historical controls and 2) the resorption rate of the concurrent controls was unusually low and fell below the range of historical control.
- Phase II: No effects attributed to treatment were observed in the reproductive parameters made at the time of necropsy.
Dead fetuses:
no effects observed
Changes in pregnancy duration:
effects observed, treatment-related
Description (incidence and severity):
- Phase I: One rabbit from the 1000 mg/kg bw/day dose group showed signs of early termination of pregnancy and was sent for a complete necropsy on gestation Day 22. This rabbit incurred a severe weight loss (673 grams) during the treatment period (Days 7 through 20) which may have contributed to the early delivery of the fetuses.
- Phase II: One rabbit from the 1000 mg/kg bw/day dose group started delivering just prior to necropsy.
Changes in number of pregnant:
not examined
Other effects:
not examined
Dose descriptor:
NOAEL
Effect level:
180 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
clinical signs
body weight and weight gain
number of abortions
Abnormalities:
no effects observed
Fetal body weight changes:
no effects observed
Reduction in number of live offspring:
no effects observed
Changes in sex ratio:
effects observed, non-treatment-related
Description (incidence and severity):
- Phase I: The distribution of male and female fetuses within the 180 mg/kg/day dose group was statistically different from that of a binomial distribution. This deviation from a binomial distribution was considered to be a random event in view of the lack of an effect on the sex ratio in fetuses from dams given 538 or 1000 mg/kg/day.
-Phase II: The distribution of male and female fetuses in the 538 mg/kg/day dose group was significantly different from that of a binomial distribution. This deviation was not seen at the high dose and hence, was considered to be a random event and not due to treatment.
Changes in litter size and weights:
not examined
Changes in postnatal survival:
not examined
External malformations:
effects observed, non-treatment-related
Description (incidence and severity):
- Phase II: In the control group a cleft palate was observed one fetus.
Skeletal malformations:
effects observed, non-treatment-related
Description (incidence and severity):
- Phase I: One fetus in the high dose group had a hemivertebrae.
- Phase II: In the 54 mg/kg bw/day dose group one fetus had a thoracic hemivertebra and an extra lumbar rib.
In the 180 mg/kg bw/day dose group one fetus had a thoracic hemivertebra.
In the high dose group a cervical and a thoracic hemivertebrae and a fused rib was seen one fetus. Another fetus had fused caudal vertebra.
Visceral malformations:
effects observed, non-treatment-related
Description (incidence and severity):
- Phase I: There was a slightly higher incidence of retroesophageal right subclavian artery in rabbits given 1000 mg/kg bw/day, suggestive of a treatment-related effect. However, this malformation has been noted in low numbers in the historical control rabbits used in the test laboratory. There were no statistically or biologically significant treatment-related differences in the incidence of any fetal variation or malformation, or in the number of malformed fetuses in any dose group.
In the control dose group, 7 fetuses had single malformations. Three fetuses had single malformations consisting of an omphalocele, missing gall bladder or ectopic kidneys. Four fetuses had a missing caudal lobe of the lung.
The 180 mg/kg bw/day dose group had 12 fetuses with malformations. Of the 12 fetuses, 11 had incidences of a missing caudal lobe of the lung. The twelfth fetus had a missing caudal lobe of the lung and a retroesophageal right subclavian artery arising from the descending aorta.
In the 538 mg/kg/ day dose group, there were 6 fetuses that were malformed. Among the malformed fetuses, three had a missing caudal lobe of the lung. The remaining three fetuses had single malformations consisting of either an omphalocele, retroesophageal right subclavian artery or an enlarged right atrium.
The high dose group had 10 fetuses with malformations. Two fetuses had retroesophageal right subclavian arteries, one of which arose from the descending aorta. A third fetus had a missing caudal lung lobe, missing cartilaginous rings in the trachea and a persistent right fourth aortic arch. A fourth fetus had a missing caudal lobe of the lung. The remaining 6 fetuses had a missing caudal lobe of the lung.
- Phase II: A total of 3,5,3,9 and 9 fetuses were malformed in the 0, 54, 180, 538 and 1000 mg/kg bw/day dose groups, respectively. However, each malformation within a dose group occurred at a low frequency or was within the historical control range observed in the test laboratory. The following visceral malformations were observed:
In two fetuses of the control group a caudal lobe of the lung was missing.
The 54 mg/kg bw/day dose group had 4 fetuses that were malformed. They each had a single malformation consisting of missing caudal lobe of the lung, retroesophageal aortic arch, ectopic kidney or persistent truncus arteriosus with no connection to the right ventricle.
In the 180 mg/kg bw/day dose group, three malformed fetuses. One fetus had a missing caudal lobe of the lung. The remaining two fetuses had retroesophageal right subclavian arteries.
Of the 8 fetuses that were malformed in the 538 mg/kg bw/day dose group, five had a missing caudal lobe of the lung. A sixth fetus had an omphalocele. A retroesophageal right subclavian artery was seen in the seventh fetus. A eighth fetus had a diaphragmatic hernia and hypoplastic lungs.
The high dose group had nine fetuses that were malformed. A missing caudal lobe of the lung was seen in six fetuses. A seventh fetus had an omphalocele with secondary constriction of the liver, liver hemorrhage and a missing caudal lobe of the lung.
Other effects:
no effects observed
Details on embryotoxic / teratogenic effects:
All fetal the malformations and variations within the treated groups were either not dose-related, occurred at low frequencies, or fell within the range of New Zealand White rabbit historical control data generated in this laboratory.
Sex:
male/female
Remarks on result:
not determinable due to absence of adverse toxic effects
Abnormalities:
effects observed, non-treatment-related
Localisation:
visceral/soft tissue: respiratory system
Developmental effects observed:
yes
Lowest effective dose / conc.:
538 mg/kg bw/day
Treatment related:
yes
Relation to maternal toxicity:
developmental effects as a secondary non-specific consequence of maternal toxicity effects
Dose response relationship:
yes
Relevant for humans:
no

Given the unusually low resorption rate observed in the concurrent control rabbits and the historical control incidence of retroesophageal right subclavian artery, the apparent increased incidence of embryonal/fetal resorptions and retroesophageal right subclavian artery observed at the high dose were interpreted to be unrelated to treatment. However, the fact that these parameters were elevated above the concurrent controls raised some concern regarding the cause of the apparent increases. Therefore, in order to improve the confidence in the conclusion that the apparent increases in resorption rates and retroesophageal right subclavian arteries were unrelated to treatment, the study was repeated (Phase II).

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
test procedure in accordance with national standard methods with acceptable restrictions
Justification for type of information:
ADAPTATION ACCORDING TO REACH ANNEX XI, section 1 - see justifications attached to IUCLID section 13.2.
- Justification WoE_RDT_Reprotox_Feb2019
- Justification RA_CAS 122-20-2_Dev.tox.2ndspecies_Nov2018
Principles of method if other than guideline:
Female onn-pregnant rabbits were administered the test substance orally by gavage for 21 days, after blood sampling all animals will be sacrificed and examined.
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH / Charles River Laboratories, France
- Housing: single
- Diet (e.g. ad libitum): Pelleted Kliba maintenance diet rabbit and guinea pig “GLP”, Provimi Kliba SA, Kaiseraugst/Switzerland; ad libitum
- Water (e.g. ad libitum): drinking water, ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-22
- Humidity (%): 30-70
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
Dose: 1000 mg/kg bw
Duration of treatment / exposure:
21 days
Frequency of treatment:
daily
No. of animals per sex per dose:
Control group: 3 animals
Test group (1000 mg/kg bw): 5 animals (due to the premature dead of 2 animals (1 found dead and 1 sacrificed after gavage error) from test group 1 (1000 mg/kg bw/d) 2 animals will be added)
Control animals:
yes, concurrent no treatment
Details on study design:
The test substance was administered to the animals orally by gavage for 21 days. For this purpose, disposable syringes (e.g. B. Braun Melsungen AG, Melsungen, Germany) and suitable gavage tubes (e.g. Nelaton soft rubber tubes, Willy Ruesch GmbH, Kernen, Germany) were used. During the study, all animals were observed for any clinically abnormal signs. One day after the last administration blood samples were obtained from all surviving animals from the ear veins. After blood sampling all animals were sacrificed and examined.
Maternal examinations:
Mortality: A check for moribund and dead animals was made twice daily from Mondays to Fridays and once daily on Saturdays, Sundays and public holidays.
Clinical signs: A cageside examination was conducted at least once daily for any signs of morbidity, pertinent behavioral changes and/or signs of overt toxicity. During the administration period all animals were checked daily for any abnormal clinically signs before the administration as well as within 5 hours after the administration. Abnormalities and changes were documented for each animal.
Food consumption: Food consumption was recorded daily for days -4 - 0 and 0 - 21.
Body weight: Body weights was recorded on day 0, 2, 4, 6, 9, 11, 14, 16, 20 and 21.
Post-Mortem Examinations:The surviving animals were sacrificed by an intravenous injection of pentobarbital (dose: 2 mL/animal). After exsanguination the animals were necropsied and examined according to 3.12. Animals which died intercurrently or were sacrificed in a moribund state were necropsied in the same way as soon as possible after their death (with the exception of the organ weights).
Clinical Pathology: Blood samples were taken from the ear veins. The following parameters were examined: Hematology: Leukocytes, Erythrocytes, Hemoglobin, Hematocrit, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), platelets, differential blood count, reticulocytes. Clinical chemistry: Alanine aminotransferase, Aspartate aminotransferase, Alkaline phosphatase,serum γ-glutamyl transferase, Inorg. phosphate, Calcium, Urea, Creatinine, Glucose, total bilirubin, total protein, Albumin, Globulins, Triglycerides, Cholesterol.
Pathology:
Necropsy: The sacrificed animals were necropsied and assessed by gross pathology. All animals which died intercurrently or were sacrificed in a moribund state were necropsied as soon as possible after their death and assessed by gross pathology.
Organ weights: The following weights were determined in all does sacrificed on schedule: Adrenal glands, kidneys, liver and spleen. The final body weights (ToxLims-System) will be transferred to the ACOPAT-System to calculate the relative organ weights.
Organ / tissue fixation: The following organs or tissues were fixed in 4% buffered formaldehyde solution: All gross lesions, adrenal glands, kidneys, liver and spleen.
Statistics:
Dunnett-test (two-sided)
Kruskal-Wallis and Wilcoxon test
Details on results:
The test substance was administered dissolved in drinking water for 21 days to groups of 3 – 5 female New Zealand White rabbits, via daily gavage. The dose levels were 0 and 1000 mg/kg body weight/day.
In the dose group (1000 mg/kg bw/d) all 5 females had reduced food consumption and lost body weight throughout the study. These animals also had reduced feces or diarrhea. Two rabbits died on the 3rd and 5th treatment day, respectively. The deceased animals showed no other pre-terminal clinical signs of toxicity, nor were there any specific necropsy findings.

Female non-pregnant rabbits were exposed to the test substance.

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available
Additional information

So far, no PNDTS is available for MIPA. However, in line with Annex XI of the REACH regulation, a registrant may propose to adapt the required standard information, including a detailed justification for this adaptation. For MIPA a PNDTS on one species (according to OECD 414) is proposed by the registrant, in order to further support the available weight of evidence approach for toxicity to reproduction. Based on the available reliable data, it is concluded, that MIPA does not induce developmental toxicitx and/or teratogenicity (see IUCLID section 13.2: "Justification_WoE_RDT_Reprotox_Feb2019").

Weight of evidence approach

In a combined repeated dose oral toxicity study with a reproduction/developmental screening study (according to OECD guideline 422 and under GLP), Wistar rats (12/sex/dose) were administerd the hydrochloride salt of MIPA (CAS no. 7780 -04 -3) 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. Pups were sacrificed on day 4 post partum and examined macroscopically for external and visceral findings. No adverse effects were observed in pup number, status at delivery, viability/mortality, sex ratio, clinical observations, body weight, and necropsy observations in any of the groups. Based on the mild anemic process in males, the NOAEL for systemic toxicity is 300 mg/kg bw/day. Thus, a NOAEL of 1000 mg/kg bw/day (the highest dose tested) was established for developmental toxicity.

No adverse effects were observed in the OECD 414 PNDTS in rats with the structural analogues DIPA and TIPA up to the limit dose. Furthermore, developmental toxicity data for TIPA in the second species rabbit (OECD414 with Picloram TIPA salt and sub-chronic dose range finder with TIPA) also revealed no adverse developmental effects but showed, that there is a MTD (maximal tolerable dose) for the dams below the limit dose.

Mode of Action Analysis / Human Relevance Framework

No information available.

Justification for classification or non-classification

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008, as amended for the tenth time in Regulation (EU) No 2017/776. As a result the substance is not considered to be classified for reproductive toxicity under Regulation (EC) No. 1272/2008.