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Key value for chemical safety assessment

Effects on fertility

Description of key information

For TIPA, CAS no. 122-20-3, a one-generation reproductive study (according to US FDA guidelines) is available. However, an EOGRTS is not available and the endpoint reproductive toxicity (fertility) is assessed in a weight of evidence approach from the available toxicological data. According to REACH annex XI, section 1 a testing does not appear scientifically necessary, using the available toxicological data with TIPA and two analogue substances DIPA, CAS 110-97-4 and MIPA, CAS 78-96-6.
The one-generation reproductive toxicity study used rats which were administered TIPA in the diet for 5 weeks prior to mating, during mating, gestation, lactation, and for 90 days post-weaning (offspring). The NOAEL for reproductive effects was reported 609 mg/kg bw/day for males and 700 mg/kg bw/day for females, the highest dose tested. In addition, under the conditions of an OECD 422 reproduction/developmental toxicity screening test with hydrochloride salt of MIPA, the NOAEL for reproductive performance, fertility and developmental toxicity is 1000 mg/kg bw/day for rats, the highest dose tested. Furthermore, repeated dose toxicity studies with TIPA in dogs and rats revealed no effects on the reproductive organs up to the limit doses.

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_WoE_Reprotox_Aug2019".
Reason / purpose for cross-reference:
read-across: supporting information
Remarks:
justification
Qualifier:
according to guideline
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:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system 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 39.7 and 43.7 mg/kg bw/day for males and females of the F1 generation, respectively)
Dose / conc.:
2 000 ppm (nominal)
Remarks:
in diet (equivalent to 160 and 182 mg/kg bw/day for males and females of the F1 generation, respectively)
Dose / conc.:
7 500 ppm (nominal)
Remarks:
in diet (equivalent to 609 and 700 mg/kg bw/day 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 602 mg/kg bw/day for males; 693 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)
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)
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 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 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:
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
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_WoE_Reprotox_Aug2019".
Reason / purpose for cross-reference:
reference to same study
Reason / purpose for cross-reference:
read-across: supporting information
Remarks:
justification
Qualifier:
according to guideline
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
Justification for study design:
This study entry for an OECD422 study with the test substance MIPA-HCl (CAS no. 7780-04-3) is part of the WoE approach for this endpoint and the justification for this WoE is attached to IUCLID chapter 13.2. Thus, a "specification of the study design for an EORTS with justifications" was not done.
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 or test system and environmental conditions:
- Source: Charles River Laboratories, Research Models and Services, Germany Gmbh
- Age at study initiation: 11-13 weeks
- Weight at study initiation: 267.4-309.0 g (males) and 186.7-217.7 g (females)
- Number of animals: 96 (12 per sex per dose group)
- Housing: Individually in type DK III stainless steel wire mesh cages supplied by Becker & Co., Castrop-Rauxel, Germany (floor area of about 800 cm2), with the following exceptions: for the overnight mating the females were put into the cages of the males
- 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 CHEMISTRA:
- 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

For TIPA, CAS no. 122-20-3, a one-generation reproductive study (according to US FDA guidelines) is available. However, an EOGRTS is not available and the endpoint reproductive toxicity (fertility) is assessed in a weight of evidence approach from the available toxicological data. Aaccording to REACH annex XI, section 1 a testing does not appear scientifically necessary, because there is sufficient weight of evidence from the available toxicological data with TIPA and two analogue substances DIPA, CAS 110-97-4 and MIPA, CAS 78-96-6.

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 (approximately 0, 40, 160 or 609 mg/kg bw/d in males or 0, 44, 182 or 700 mg/kg bw/d in females) for 5 weeks prior to mating as well as during mating, gestation and lactation [1988; RL2]. 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 of pups/litter, litter size) were noted. The NOAEL was reported to be 609 mg/kg bw/d for males and 700 mg/kg bw/d for females, the highest doses tested.

In a combined repeated dose toxicity study with the reproduction/developmental toxicity screening test [2008, RL1], rats were exposed to a 65.4 % aqueous MIPA HCl solution with 100, 300 and 1000 mg/kg bw/d, corresponding to 67, 202 and 673 mg/kg bw/d organic MIPA. Only slightly reduced hemoglobin and hematocrit values were observed in the high dose male rats after 38 days of exposure, indicating a mild anemic process. No toxicity effects were observed in female rats after 45 days of exposure at all dose levels nor in the F1 pups and no developmental and fertility effects were observed. Based on the mild effects in the high dose male rats, the NOAEL for general, systemic toxicity of the test substance was set at the mid dose level (i.e. 202 mg/kg bw/d expressed as organic MIPA).

Additionally, repeated dose toxicity studies in beagle dogs and rats with TIPA revealed no effects on the reproductive organs up to the limit doses of 272 mg/kg bw/d and 1200 mg/kg bw/d, respectively.

Please refer to the justification for adaptation according to REACH annex XI, section 1; attached to IUCLID section 13.2: “Justification_WoE_Reprotox_Aug2019”.

Effects on developmental toxicity

Description of key information

In an OECD414 developmental toxicity study, pregnant female rats were administered TIPA orally per gavage on gestation days 6 to 15. The maternal and developmental NOAELs were 400 and >1000 mg/kg bw/d, respectively. In a second OECD414 study, pregnant female rabbits were administered picloram TIPA salt orally per gavage on GD7 -19. The maternal and developmental NOAELs were 180 and 1000 mg/kg bw/d, respectively (corresponding to 80 and 442 mg/kg bw/d of TIPA, respectively).

Link to relevant study records

Referenceopen allclose all

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
27 Jan 1994 - 21 Feb 1994
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
GLP compliance:
yes
Limit test:
no
Specific details on test material used for the study:
- 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 or test system 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/100 mL, 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:
- conception rate (%) = number of pregnant animals / number of fertilized animals x 100.
- preimplantation loss (%) = (number of corpora lutea - number of implantations) / number of corpora lutea x 100.
- postimplantation loss (%) = (number of implantations - number of live fetuses) / number of implantations x 100.
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.
Historical control data:
Yes.
Clinical signs:
no effects observed
Description (incidence and severity):
There were no abnormal clincial findings in any dam of anyone group.
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Description (incidence):
There were no mortalities in any of the groups.
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. 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. For further details see attached document (result tables).
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, which is related to the test substance administration.
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):
The uterus weights of the animals were not influenced by the administration of the test substance. The differences between the groups are without biological relevance. For further details see attached document (result tables).
Gross pathological findings:
no effects observed
Description (incidence and severity):
At necropsy, single animals of the control and all substance treated groups showed lungs with edema and/or marginal emphysema; these findings, which showed no relation to dosing, have to be related to the sacrifice of the animals.
- number (%) of dams with lung edema in 0, 100, 400, 1000 mg/kg bw/d group: 9, 4, 5, 6 (36, 16, 20, 24 %)
- number (%) of dams with marginal lung emphysema in 0, 100, 400, 1000 mg/kg bw/d group: 3, 2, 2, 2 (12, 8, 8, 8 %).
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
Description (incidence and severity):
For further details see attached document (result tables).
Pre- and post-implantation loss:
no effects observed
Description (incidence and severity):
For further details see attached document (result tables).
Total litter losses by resorption:
no effects observed
Description (incidence and severity):
For further details see attached document (result tables).
Early or late resorptions:
no effects observed
Description (incidence and severity):
For further details see attached document (result tables).
Dead fetuses:
no effects observed
Description (incidence and severity):
For further details see attached document (result tables).
Changes in pregnancy duration:
no effects observed
Changes in number of pregnant:
no effects observed
Description (incidence and severity):
The conception rate varied between 92% (test group 3 - 1,000 mg/kg body weight/day) and 100% (control group and test group 2 - 400 mg/kg body weight/day). For further details see attached document (result tables).
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 (total, early, late) 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. For further details see attached document (result tables).
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):
The mean fetal weights in test groups 1, 2 and 3 (100, 400 and 1,000 mg/kg body weight/day) were not influenced by the test substance administration and there were no biologically relevant differences concerning the means of the control group and of the substance-treated groups. For further details see attached document with result tables.
Reduction in number of live offspring:
no effects observed
Description (incidence and severity):
There were no substance-related and/or biologically relevant differences between the groups in the number of viable fetuses. For further details see attached document with result tables.
Changes in sex ratio:
no effects observed
Description (incidence and severity):
The sex distribution of the fetuses in test groups 1 - 3 (100, 400 and 1,000 mg/kg body weight/day) was comparable with the control fetuses. The differences observed in comparison to the control are without any biological relevance. For further details see attached document with result tables.
Changes in litter size and weights:
not examined
Changes in postnatal survival:
not examined
External malformations:
no effects observed
Description (incidence and severity):
There are no statistically significant differences between the control and the substance-treated groups with respect to external malformation, variation and/or retardation which could be related to the test substance administration.

The external examination of the fetuses revealed two malformations in one fetus of test group 2 (400 rng/kg body weight/day). This fetus (No . 4 of animal No. 71) showed anophthalmia on the right side and a microphthalmia of the left eye. Due to the missing dose-response relationship and the isolated nature of this finding, it is considered tobe spontaneous in nature. Moreover, this finding can also be found at a low incidence in the historical control data from the breeder. No external variations were found in any group. Only one so-called unclassifiad observation (placentae fused) was recorded for one fetus of test group 0 (0 mg/kg body weight/day) and one fetus of test group 1 (100 mg/kg body weight/day); a finding without dose-response relationship and thus considered not relevant.

For further details see attached documents (result tables and historical control data).
Skeletal malformations:
effects observed, non-treatment-related
Description (incidence and severity):
There are no statistically significant differences between the control and the substance-treated groups with respect to skeletal malformation, variation and/or retardation which could be related to the test substance administration.

The few statistically significant differences which occurred were exclusively related to fetal skeletal variations and retardations and 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 body weight/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 (see respective tables below).

Various malformations of the sternum and the vertebral column were seen in the control and the three dose groups, occurring without any statistically significant differences between the substance-treated groups and the concurrent control group: 0, 100, 400, 1000 mg/kg bw/d: 8/177, 8/176, 6/193, 15/178 fetuses; 8/25, 5/25, 5/25, 10/23 litters. Furthermore, all of the aforementioned skeletal malformations or very similar ones can be found at comparable or even higher fetal/litter incidences in the historical control data.
The variations elicited, which were related to the ribs, the sternum, the skull and the vertebral column, appeared without a clear dose-response relationship, can be found in a similar frequency in the historical control data and/or the differences between the groups are without biological relevance.

For further details see attached documents (result tables and historical control data).
Visceral malformations:
no effects observed
Description (incidence and severity):
There are no statistically significant differences between the control and the substance-treated groups with respect to soft tissue malformation, variation and/or retardation which could be related to the test substance administration.

The examination of the organs of the fetuses revealed three different malformations. Hydrocephaly was recorded for one fetus of test group 2 (400 mg/kg body weight/day) (No. 4 of animal No. 71), truncus arteriosus communis was seen in one fetus of test group 1 (100 mg/kg body weight/day) (No. 2 of animal No. 49) and dilatation of both heart ventricles (globular shaped heart) occurred in three fetuses; in one fetus of test group 0 (No. 10 of animal No. 16), in one fetus of test group 2 (No. 4 of animal No. 71) and in one fetus of test group 3 (No. 12 of animal No. 80). Two of these malformations (hydrocephaly and dilatation of both ventricles) are also present at a low incidence in the historical control data. The isolated and disparate nature of the three soft tissue malformations and the missing dose-response relationship does not suggest any treatment-related aetiology; therefore, these malformations are considered tobe spontaneous in nature. Variations (dilated renal pelvis and/or hydroureter) were detected in all groups without any statistically significant and/or biologically relevant differences between the groups. Both findings are very common ones in the rat strain used and all respective values are fully in the range of biological variation. No so-called unclassified observations (like bloody imbibition of kidney(s)) were recorded during the soft tissue examination.

For further details see attached documents (result tables and historical control data).
Other effects:
no effects observed
Description (incidence and severity):
The mean placental weights in test groups 1, 2 and 3 (100, 400 and 1,000 mg/kg body weight/day) were not influenced by the test substance administration and were similar to the control values. For further details see attached document (result tables).
Key result
Dose descriptor:
NOEL
Effect level:
1 000 mg/kg bw/day
Based on:
test mat.
Remarks on result:
other: There were no (adverse toxic) effects observed up to the highest dose tested.
Key result
Abnormalities:
no effects observed
Key result
Developmental effects observed:
no

For further details see attached documents (result tables and historical control data).

Endpoint:
developmental toxicity
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Justification for type of information:
According 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), further testing for developmental toxicity in a 2nd species does not appear scientifically necessary due to the read across to a structural similar substance. Furthermore, the available data are adequate to support the non-classification for developmental toxicity and the risk assessment. See read-across justification in IUCLID section 13.2. "Justification_RA-Dev.tox_2ndspecies_Aug2019"
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across: supporting information
Remarks:
RA justification
Reason / purpose for cross-reference:
read-across: supporting information
Remarks:
experimental study
Reason / purpose for cross-reference:
read-across: supporting information
Remarks:
data evaluation report
Dose descriptor:
NOAEL
Effect level:
80 mg/kg bw/day
Based on:
test mat.
Remarks:
calculated with molecular weight of TIPA = 191.27 g/mol and of Picloram TIPA salt = 432.73 g/mol.
Basis for effect level:
body weight and weight gain
clinical signs
number of abortions
Dose descriptor:
NOEL
Effect level:
440 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no developmental toxicity up to the highest dose tested
Developmental effects observed:
no
Treatment related:
no

TIPA doses were calculated based on the molecular weight of TIPA = 191.27 g/mol and of Picloram TIPA salt = 432.73 g/mol.

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

Twenty-five female Wistar rats were administered TIPA at 0, 100, 400 or 1000 mg/kg bw/day via oral gavage on gestation days 6 to 15 in an OECD guideline 414 study (1995, RL 1). Food consumption, body weights and clinical signs of the dams were recorded. At necropsy on day 20 post coitum, dams were investigated for gross pathology (including weight determination of the unopened uterus), fetuses were removed and sexed, weighed and further investigated for any external, soft tissue and/or skeletal findings. In the highest dose group, decreased food consumption and reduced body weight gain was observed in the dams. No further treatment-related effects were observed in the dams or in the fetuses. Thus, NOAELs of 400 mg/kg bw/day for maternal toxicity and 1000 mg/kg bw/day (the highest dose tested) for teratogenicity were established.

A second study with rabbits is available performed with picloram TIPA salt (a structural analogue of TIPA, see attached read-across justification in IUCLID chapter 13.2: “Justification_RA_Dev.tox_2ndspecies_Aug2019”). In a GLP compliant study, performed according to OECD guideline 414, the maternal, embryonal and fetal toxicity and teratogenicity of picloram TIPA salt was assessed (1992, RL 2). The study was conducted in two phases. In Phase I, groups of 18 inseminated New Zealand White rabbits were administered the test substance by oral gavage on GD 7 -19 at targeted dose levels of 0, 180, 538 or 1000 mg/kg bw/d. Phase II of this study was conducted as a repeat of Phase I to determine whether the increased resorption rate and certain fetal observations in Phase I were reproducible and/or attributed to treatment. In Phase II, an additional dose group of 54 mg/kg bw/d was added at the low end to establish a no-observed-effect level. On GD28 all surviving animals were necropsied and liver, kidneys and gravid uterine weights were recorded. The ovaries were examined for corpora lutea and the uterus was examined for implantations and resorptions. Fetuses were removed, weighed, sexed and examined for external, visceral and skeletal alterations. Maternal toxicity was observed in rabbits given 538 or 1000 mg/kg bw/d, as evidenced by decreased feed consumption accompanied by higher incidences of decreased feces, soft feces, perineal soiling and decreased body weight gain (Phase I and II). In addition, an increased incidence of high-dose dams aborting (one in Phase I and two in Phase II) was also interpreted to represent maternal toxicity. No other adverse maternal or embryonal/fetal effects attributed to treatment were observed at any dose level. The test substance was not teratogenic at any dose level tested. In conclusion, the maternal no-observed-adverse-effect level (NOAEL) was 180 mg/kg bw/day. Corrected for molecular weight this is equivalent to 80 mg/kg bw/day TIPA. The no-observed-effect level (NOEL) for embryonal/fetal toxicity or teratogenicity of the test substance was 1000 mg/kg/d, the highest dose level tested and equivalent to 440 mg/kg bw/d TIPA.

The limit dose for an OECD 414 study that needs to be tested is 1000 mg/kg bw/d. Based on this there might be remaining uncertainties for hazard characterisation. To further clarify these uncertainties, a supporting shortterm repeated dose study was performed with New Zealand White rabbits (2016, RL 2): TIPA was administered dissolved in drinking water for 21 days to groups of non-pregnant 35 female New Zealand White rabbits, via daily gavage. Animals were exposed to either 0 or 1000 mg/kg bw/d. In the dose group (1000 mg/kg bw/d) all 5 females showed reduced food consumption and lost body weight throughout the study. These animals also had reduced faeces or diarrhoea. Two rabbits died on the 3rd and 5th treatment day, respectively. The deceased animals showed no other preterminal clinical signs of toxicity, nor were there any specific necropsy findings. In conclusion, if additional testing would be performed with TIPA, the doseselection for this study would be in the range of 500 mg/kg bw/d (half of the lethal dose), which is comparable to the converted dose of 440 mg TIPA/kg bw/d from the OECD 414 rabbit study with picloram TIPA salt (where no developmental toxic effects were observed).

This is also discussed in the read-across justification, attached to IUCLID section 13.2: “Justification_RA_Dev.tox_2ndspecies_Aug2019”.

Mode of Action Analysis / Human Relevance Framework

No further details.

Justification for classification or non-classification

Based on the available studies, there is no indication that TIPA can cause effects on fertility or developmental toxicity. Therefore, TIPA does not need to be classified according to the EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008.

Additional information