3-{2-[2-(3-azaniumylpropoxy)ethoxy]ethoxy}propan-1-aminium di[(2Z)-3-carboxyprop-2-enoate]

Endpoint summary

Description of key information

Repeated dose toxicity: oral. 33Oxy.001

Under the conditions of the study, the NOAEL has been determined as 600 mg/kg bw/day in relation to parental systemic toxicity (male/female); 100 mg/kg bw/day in relation to local effect (female – histopathology (lesions in the upper digestive tract at all dose levels) and < 100 mg/kg bw/day in relation to local effect (male – histopathology (lesions in the upper digestive tract at all dose levels).

Repeated dose toxicity: oral. Maleic Acid_RA to Maleic Anhydride.002

The NOEL of a 90-days dietary feeding study with Maleic Anhydride to Beagle dogs was 60 mg/kg/day, which was the highest concentration tested.

Repeated dose toxicity: oral. Maleic Acid_RA to Maleic Anhydride.003

The LOEL of a 2-years dietary feeding study with rats was 32 mg/kg/day, the NOEL was 10 mg/kg/day.

Repeated dose toxicity: oral. Maleic Acid.004

Toxic effects were observed at concentrations of 0.5 %, 1 % and 1.5 % of maleic acid in the diet, fed to male rats for two years. The main findings were increased mortality rate, reduced body weight/body weight gain and histopathological changes in liver, testis and kidneys.

Repeated dose toxicity: oral. Maleic Acid_RA to Maleic Anhydride.005

The NOEL of a 90-days dietary feeding study with rats was 40 mg/kg/day, which was the highest concentration tested.

Repeated dose toxicity: oral. Maleic Acid_RA to Maleic Anhydride.006

The LOEL of a 90-days dietary feeding study with rats was 100 mg/kg/day, which was the lowest concentration tested.

Repeated dose toxicity: oral. Maleic Acid_RA to Maleic Anhydride.007

The LOEL of a 183-days dietary feeding study with rats was 250 mg/kg/day, which was the lowest concentration tested.

Key value for chemical safety assessment

Toxic effect type:

dose-dependent

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Reference 1

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Remarks:

Study has been performed on Bis-Aminopropyl Diglycol aka 3,3'-oxybis(ethyleneoxy)bis(propylamine); (EC 224-207-2; CAS 4246-51-9). This is being used as the read-across substance to Bis Aminopropyl Diglycol Dimaleate; (EC 818-033-1; CAS 1629579-82-3).

Adequacy of study:

weight of evidence

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study conducted in accordance with GLP.

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: EPA, Health Effects Test Guidelines; OPPTS 870.3650: Combined Repeated Dose Toxicity Study With the Reproduction/Developmental Toxicity Screening Test

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS

- Age at supplied: 10-11 weeks
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH, Sulzfeld, Germany
- Weight at study initiation: Males: 321 - 322 g, Females 205 - 209 g
- Fasting period before study: no
- Housing: individually in Makrolon type M III cages;
- Exceptions:
- During overnight matings, male and female mating partners were housed together in Makrolon type M III cages
- Pregnant animals and their litters were housed together until PND 4 (end of lactation).
- For motor activity (MA) measurements the animals were housed individually in polycarbonate cages (floor area of about 800 cm2) and
small amounts of bedding material
- The cages with the test animals were arranged on the racks in such a way that uniform experimental conditions (ventilation and light) were ensured.
- Diet, ad libitum: Ground Kliba maintenance diet mouse-rat “GLP” meal, supplied by Provimi Kliba SA, Kaiseraugst, Switzerland
- Water, ad libitum: drinking water
- Acclimation period: On the day of arrival the animals were subjected to an appropriate acclimatization period.

ENVIRONMENTAL CONDITIONS
- Temperature: 20-24°C
- Humidity: 30-70%
- Air changes (per hr): 15
- Photoperiod: 12-hour light/12-hour dark cycle

IN-LIFE DATES: From: 2012-01-09 To: 2013-03-12

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:
The appropriate amount of test substance was weighed out depending on the desired concentration. Then, drinking water was filled up to the desired volume, subsequently released with a magnetic stirrer. During administration of the test substance, preparations were kept homogeneous by stirring with a magnetic stirrer. The test substance preparations were produced at least once a week and were stored at room temperature.

VEHICLE
- Justification for use and choice of vehicle: solubility
- Concentration in vehicle:
1.00 mg/100 mL (100 mg/kg bw/d)*, 3.00 mg/100 mL (300 mg/kg bw/d)*, 10.00 and 6.00 mg/100 mL (1000 and 600 mg/kg bw/d)*
*) The dose refers to the body weight of the individual rats determined most recently.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The analyses of the test-substance preparations were carried out at the Analytical Chemistry Laboratory of Experimental Toxicology and Ecology of BASF SE, Ludwigshafen, Germany.
The stability of the test substance in drinking water for a period of 7 days at room temperature was proven during the study (BASF project No. 01Y0401/11Y018; see PART III, Supplement).
Homogeneity and concentration control analyses of the test-substance preparations were performed in all concentrations at the start of the administration period. Additionally, samples from all concentrations as reverse samples for concentration control analysis were taken at the end of the study.

Duration of treatment / exposure:

After the acclimatization period, the test substance was administered orally via gavage to the F0 generation parental animals, daily at the same time in the morning (exception: no administration to animals being in labor). The treatment lasted up to one day prior to sacrifice. The animals of the control group were treated in the same way with the vehicle only (drinking water).

Frequency of treatment:

Daily

Remarks:

Doses / Concentrations:
100, 300 and 1000 (reduced to 600 on study day 7) mg/kg bw/d
Basis: Actual ingested

No. of animals per sex per dose:

10 animals

Control animals:

yes, concurrent vehicle

Details on study design:

The oral route was selected since this was proven to be suitable for the detection of a toxicological hazard.

F0 generation parental animals and their progeny
The animals of the control group were treated in the same way with the vehicle only (drinking water). The calculation of the administered volume was generally based on the most recent individual body weights. Fourteen days after the beginning of treatment, males and females from the same test group were mated overnight in a ratio of 1:1 (details of pairing see 3.7.2.). On study day 49, a functional observational battery and motor activity measurement were carried out in the first five surviving parental male animals per group. The females were allowed to litter and rear their pups until day 4 after parturition. On PND 4, all pups were sacrificed and examined. On study day 56, a functional observational battery and motor activity measurement was carried out in the first five surviving parental female animals (with litter) per group. From the first 5 surviving parental male animals per group and the first 5 surviving parental female animals which delivered first urinalysis was carried out on study days 51 (males) and 58 (females). Clinicochemical and hematological examinations were carried out on study days 60 (males) and 63 (females). At the end of the study (males: study day 60, females: study day 63), the animals were sacrificed after a fasting period (withdrawal of food) for at least 16-20 hours.

Age at mating of the mated animals in the study: 13-14 weeks.

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
A cageside examination was conducted at least once daily for any signs of morbidity, pertinent behavioral changes and signs of overt toxicity. Abnormalities and changes were documented daily for each affected animal.
The littering and lactation behavior of the dams was generally evaluated in the mornings in combination with the daily clinical inspection of the dams. Only particular findings (e.g. inability to deliver) were documented on an individual dam basis.
On weekdays (except public holidays) the parturition behavior of the dams was inspected in the afternoons in addition to the evaluations in the mornings.
The day of littering was considered the 24-hour period from about 15.00 h of one day until about 15.00 h of the following day.

DETAILED CLINICAL OBSERVATIONS: Yes
Detailed clinical observations (DCO) were performed in all animals prior to the administration period and thereafter at weekly intervals. The findings were ranked according to the degree of severity, if applicable. The animals were transferred to a standard arena (50 × 37.5 cm with sides of 25 cm high).
The following parameters were examined: abnormal behavior when handled, fur, skin, posture, salivation, respiration, activity/arousal level, tremors, convulsions, abnormal movements, impairment of gait, lacrimation, palpebral closure, exophthalmus, feces (appearance/consistency), urine, pupil size

BODY WEIGHT: Yes
Body weight was determined before the start of the administration period in order to randomize the animals. During the administration period body weight was determined on study day 0 (start of the administration period) and thereafter once a week at the same time of the day (in the morning).
The body weight change of the animals was calculated from these results.
The following exceptions are notable for the female animals:
- During the mating period the parental fe¬males were weighed on the day of positive evidence of sperm (GD 0) and on GD 7, 14 and 20.
- Females with litter were weighed on the day of parturition (PND 0) and on PND 4.
- Females without a litter and without positive evidence of sperm in the vaginal smear were weighed weekly. These body weight data were
solely used for the calculations of the dose volume.


FOOD CONSUMPTION AND COMPOUND INTAKE:
Generally, food consumption was determined once a week for male and female parental animals, with the following exceptions:
- Food consumption was not determined during the mating period (male and female F0 animals)
- Food consumption of the F0 females with evidence of sperm was determined on GD 0-7, 7-14, 14-20.
- Food consumption of F0 females, which gave birth to a litter, was determined for PND 1-4.
Food consumption was not determined in females without positive evidence of sperm (during the mating period of dams used in parallel) and females without litter (during the lactation period of dams used in parallel).

FOOD EFFICIENCY: No

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
Drinking water consumption was monitored by daily visual inspection of the water bottles for any changes in volume.

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: In the morning blood was taken from the retroorbital venous plexus from fasted animals.
The animals were anaesthetized using isoflurane (Isoba®, Essex GmbH, Munich, Germany). The blood sampling procedure and
subsequent analysis of blood and serum samples were carried out in a randomized sequence.
- Anaesthetic used for blood collection: Yes (isoflurane)
- Animals fasted: Yes
- How many animals: first 5 surviving parental males and the first 5 surviving females with litter (in order of delivery) per group
- Parameters were determined in blood with EDTA K3 as anticoagulant using a particle counter (Advia 120 model; Bayer, Fernwald, Germany)
- Parameters examined: see Table 1 in 'Any other information on materials and methods incl. tables'.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: In the morning blood was taken from the retroorbital venous plexus from fasted animals.
- Animals fasted: Yes
- How many animals: first 5 surviving parental males and the first 5 surviving females with litter (in order of delivery) per group
- An automatic analyzer (Hitachi 917; Roche, Mannheim, Germany) was used to examine the clinicochemical parameters
- Parameters examined: see Table 2 in 'Any other information on materials and methods incl. tables'.

URINALYSIS: Yes
- Time schedule for collection of urine: overnight
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes
- The dry chemical reactions on test strips (Combur 10 test M, Roche, Mannheim, Germany) used to determine urine constituents semiquantitatively
were evaluated with a reflection photometer (Miditron M; Roche, Mannheim, Germany).
- Parameters examined: see Table 3 in 'Any other information on materials and methods incl. tables'.

NEUROBEHAVIOURAL EXAMINATION: Yes
- Functional observational battery (FOB) was performed in all animals towards the end of the administration period
- Dose groups that were examined: all animals

- Battery of functions tested:
Home cage observations:
Attention was paid to: Posture, Tremors, Convulsions, Abnormal movements, Gait abnormalities.
Open field observations:
Behavior when removed from cage, fur, skin, salivation, nose discharge, lacrimation, eyes/pupil size, posture, palpebral closure, respiration, tremors, convulsions, abnormal movements, impairment of gait, activity/arousal level, feces (number of fecal pellets/appearance/consistency) within two minutes, urine (appearance/quantity) within two minutes, number of rearings within two minutes.
Sensorimotor tests/reflexes:
Approach response, touch response, vision ("visual placing response"), pupillary reflex, pinna reflex, audition ("startle response"), coordination of movements ("righting response"), behavior during "handling", vocalization, pain perception ("tail pinch"), grip strength of forelimbs, grip strength of hindlimbs, landing foot-splay test, other findings.

Motor activity (MA) measured on the same day as the FOB was performed.

Sacrifice and pathology:

SACRIFICE
- The F1 offspring was scheduled sacrifice on PND 4 under isoflurane anesthesia with CO2.
- These animals were subjected to postmortem examinations as follows: All pups were examined externally and eviscerated; their organs were assessed macroscopically. All stillborn pups and all pups that died before PND 4 were examined externally, eviscerated and their organs were assessed macroscopically.
All pups without notable findings or abnormalities were discarded after their macroscopic evaluation. Animals with notable findings or abnormalities were evaluated on a case-by-case basis, depending on the type of finding noted.

GROSS NECROPSY
- Gross necropsy consisted of external examination

HISTOPATHOLOGY / ORGAN WEIGTHS
Not performed

Other examinations:

This study was performed in an AAALAC-approved laboratory in accordance with the German Animal Welfare Act and the effective European Council Directive.

The supplier assayed the food used in the study for chemical and microbiological contaminants.
The drinking water is regularly assayed for chemical contaminants by the municipal authorities of Frankenthal and by the Environmental Analytics Water/Steam Monitoring Department of BASF SE as well as for the presence of microorganisms by a contract laboratory.
The bedding and the enrichment are regularly assayed for contaminants (chlorinated hydrocarbons and heavy metals).

 
Table 1: Hematology
Parameter
Leukocyte count (WBC)
Erythrocyte count (RBC)
Hemoglobin (HGB)
Hematocrit (HCT)
Mean corpuscular volume (MCV)
Mean corpuscular hemoglobin (MCH)
Mean corpuscular hemoglobin concentration (MCHC)
Platelet count (PLT)
Differential blood count
Reticulocytes


Table 2: Clinical chemistry (Enzyme/Blood Chemistry Parameter)
Enzyme (systematic name and system number)
Alanine aminotransferase (ALT)
(L-alanine: 2-oxoglutarate aminotransferase; EC 2.6.1.2.)
Aspartate aminotransferase (AST)
(L-aspartate: 2-oxoglutarate aminotransferase; EC 2.6.1.1.)
Alkaline phosphatase (ALP)
(orthophosphoric acid monoester phosphohydrolase; EC 3.1.3.1.)
g-Glutamyltransferase (GGT)
(g-glutamyl) peptide: aminoacid-g-glutamyl-transferase; EC 2.3.2.2.)
 
Blood Chemistry Parameter
Sodium (NA)
Potassium (K)
Chloride (CL)
Inorganic phosphate (INP)
Calcium (CA)
Urea (UREA)
Creatinine (CREA)
Glucose (GLUC)
Total bilirubin (TBIL)
Total protein (TPROT)
Albumin (ALB)
Globulins (GLOB)
Triglycerides (TRIG)
Cholesterol (CHOL)
Bile acids (TBA)


Table 3: Urinalysis
Parameter
pH
Protein
Glucose
Ketones
Urobilinogen
Bilirubin
Blood
Specific gravity
Sediment
Color, turbidity
Volume



Table 4: Histopathology

Organ samples
Test group
 
0
1
2
3
1.    All gross lesions
A2
A2
A2
A2
2.    Adrenal glands
A4
 
 
A4
3.    Bone marrow (femur)
A4
 
 
A4
4.    Brain
A4
 
 
A4
5.    Cecum
A4
 
 
A4
6.    Cervix
A1
 
 
A1
7.    Coagulation glands
A1
 
 
A1
8.    Colon
A4
 
 
A4
9.    Duodenum
A4/A3
A3
A3
A4/A3
10. Epididymides
A1
 
 
A1
11. Heart
A4
 
 
A4
12. Ileum
A4
 
 
A4
13. Jejunum
A4
 
 
A4
14. Kidneys
A4
 
 
A4
15. Liver
A4
 
 
A4
16. Lung
A4
 
 
A4
17. Lymph nodes
(mesenteric and axillary lymph nodes)
A4
 
 
A4
18. Ovaries
A1
 
 
A1
19. Oviducts
A1
 
 
A1
20. Peyer’s patches
A4
 
 
A4
21. Prostate
A1
 
 
A1
22. Rectum
A4
 
 
A4
23. Sciatic nerve
A4
 
 
A4
24. Seminal vesicles
A1
 
 
A1
25. Spinal cord
(cervical, thoracic and lumbar cords)
A4
 
 
A4
26. Spleen
A4
 
 
A4
27. Stomach
(forestomach and glandular stomach)
A1
A1
A1
A1
28. Testes
A1
 
 
A1
29. Thymus
A4
 
 
A4
30. Thyroid glands
A4
 
 
A4
31. Trachea
A4
 
 
A4
32. Urinary bladder
A4
 
 
A4
33. Uterus
A1
 
 
A1
34. Vagina
A1
 
 
A1

METHODS/SCOPE OF EXAMINATIONS: 
A
=
Hematoxylin-eosin (H&E)
1
=
all animals per test group
2
3
=
=
all affected animals per test group
all male animals per test group
4
=
5 animals per sex and test group, females with litters only, same animals as used for clinical pathology examination


--------------------------------------------
Statistics of clinical examination

- Parameter:
Food consumption (parental animals), body weight and body weight change (parental animals and pups; for the pup weights, the litter means were used), duration of gestation, number of implantation sites, postimplantation loss and % postimplantation loss, number of pups delivered per litter, viability index
- Statistical test:
Simultaneous comparison of all dose groups with the control group using the DUNNETT-test (two-sided) for the hypothesis of equal means

- Parameter:
Male and female mating indices, male and female fertility indices, gestation index, females with liveborn pups, females with stillborn pups, females with all stillborn pups, live birth index, pups stillborn, pups died, pups cannibalized
- Statistical test:
Pairwise comparison of each dose group with the control group using FISHER'S EXACT-test for the hypothesis of equal proportions

- Parameter:
Number of mating days
- Statistical test:
Pairwise comparison of the dose group with the control group using the WILCOXON-test (one-sided) with BONFERONI-HOLM-Adjustment for the hypothesis of equal medians

- Parameter:
Feces, rearing, grip strength of forelimbs and hindlimbs, landing foot-splay test, motor activity
- Statistical test:
Non-parametric one-way analysis using KRUSKAL-WALLIS test (two-sided). If the resulting p-value was equal or less than 0.05, a pairwise comparison of each dose group with the control group was performed using WILCOXON-test (two-sided) for the equal medians

Statistics of pathology
Means and standard deviations were calculated.
- In addition, the following statistical analyses were carried out:
Weight parameters
- Statistical test: 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 the WILCOXON test for the hypothesis of equal medians


Statistics of clinical pathology
Means, medians and standard deviations of each test group were calculated for several parameters.
- In addition, the following statistical analyses were carried out:

Blood parameters
- Statistical test for parameters with bidirectional changes:
Non-parametric one-way analysis using KRUSKAL-WALLIS test. If the resulting p-value was equal or less than 0.05, a pairwise comparison of each dose group with the control group was performed using WILCOXON-test (two-sided) for the hypothesis of equal medians
- Statistical test for parameters with unidirectional changes:
Pairwise comparison of each dose group with the control group using the WILCOXON-test (one-sided) for the hypothesis of equal medians

Urinalysis parameters (apart from urine color and turbidity)
- Statistical test: Pairwise comparison of each dose group with the control group using the WILCOXON-test (one-sided) for the hypothesis of equal medians

Statistics:

Please refer to 'Any other information on materials and methods incl. tables'.

Clinical signs:

effects observed, non-treatment-related

Description (incidence and severity):

CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
All groups:
No rat died prematurely in the present study.
No test substance-related, adverse findings were noted.
Reproductive Performance: No test substance-related, adverse findings were noted.
Clinical Pathology: No test substance-related, adverse findings were noted.

Mortality:

mortality observed, non-treatment-related

Description (incidence):

CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
All groups:
No rat died prematurely in the present study.
No test substance-related, adverse findings were noted.
Reproductive Performance: No test substance-related, adverse findings were noted.
Clinical Pathology: No test substance-related, adverse findings were noted.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
On study day 7 (premating) mean body weight of male animals in test group 3 (1000 and 600 mg/kg bw/d) was significantly decreased (-6%). Although not significantly altered mean body weight loss in female animals was observed after 7 days of treatment.
Mean body weight change values during premating were significantly decreased in male animals of test group (1000 and 600 mg/kg bw/d) between study days 0-7 (-84%) and 0-13  (-48%). Although not significantly altered mean body weight change value in female animals was observed between study days 0-7 days. All mentioned findings were assessed as being related to treatment.

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

FOOD CONSUMPTION AND COMPOUND INTAKE / TEST SUBSTANCE INTAKE (PARENTAL ANIMALS)
In test group 3 (1000 and 600 mg/kg bw/d) food consumption during premating was significantly decreased in male animals between study days 0-7 (-17%) and 0-13 (-11%) as well as in female animals between study days 0-7 (-19%). These findings were assessed as being related to treatment with the test substance by irritating the upper digestive tract.
No other findings were observed for male and female animals in test group 1 and 2 (100 and 300 mg/kg bw/d) nor for male and female animals of test group 3 (1000 and 600 mg/kg bw/d) after reducing the dose level.

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

effects observed, non-treatment-related

Description (incidence and severity):

WATER CONSUMPTION AND COMPOUND INTAKE
No test substance-related, adverse findings were noted.

Ophthalmological findings:

not specified

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

HAEMATOLOGY / CLINICAL CHEMISTRY / URINALYSIS
See clinical pathology

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

HAEMATOLOGY / CLINICAL CHEMISTRY / URINALYSIS
See clinical pathology

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

ORGAN WEIGHTS (PARENTAL ANIMALS)
No remarks.

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

GROSS PATHOLOGY (PARENTAL ANIMALS) / HISTOPATHOLOGY (PARENTAL ANIMALS)
Treatment of male and female Wistar rats with up to 1000 and 600 mg/kg bw/d of the test substance led to treatment-related findings in the upper digestive tract. The hyperplasia, hydropic degeneration of squamous cells and inflammatory cell infiltrates, occasionally with multinucleated giant cells in the forestomach as well as the hyperemia, eosinophilic cytoplasmic change, increased mitotic figures, submucosal edema and inflammatory cell infiltrates in the glandular stomach were regarded to be signs of a slight irritating effect of the test substance. These findings were almost exclusively observed around or at the margo plicatus and in the glandular stomach in the fundic area. They were regarded to be adverse. The increase in thickness in the duodenum was also regarded to be treatment-related and as the specific mechanism could not be determined, it was also judged to be adverse in nature. All these adverse findings described above are regarded to be a local irritating effect but no systemic toxicity.

Neuropathological findings:

no effects observed

Description (incidence and severity):

NEUROBEHAVIOUR
Home cage observations: No test substance-related effects were observed.
Open field observations: No test substance-related effects were observed.
Sensorimotor tests/reflexes: No test substance-related effects were observed.
Quantitative Parameters: No test substance-related effects were observed.
Motor activity measurement: There were no significant deviations concerning the overall motor activity (summation of all intervals) and regarding the single intervals in male and female animals of all test groups in comparison to the concurrent control group.

Histopathological findings: non-neoplastic:

not specified

Histopathological findings: neoplastic:

not specified

Other effects:

no effects observed

Description (incidence and severity):

OTHER FINDINGS (PARENTAL ANIMALS)
No findings on the reproduction tract were observed. All other findings occurred either individually or were biologically equally distributed over control and treatment groups. They were considered to be incidental or spontaneous in origin and without any relation to treatment.

Dose descriptor:

NOAEL

Remarks:

parental systemic toxicity

Effect level:

600 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Starting dose: 1000 mg/kg bw/d, reduced to 600 mg/kg bw/d on study day 7

Dose descriptor:

NOAEL

Remarks:

local effect

Effect level:

100 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

female

Basis for effect level:

other: Histopathology (lesions in the upper digestive tract at higer dose levels)

Dose descriptor:

NOAEL

Remarks:

local effect

Effect level:

< 100 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male

Basis for effect level:

other: Histopathology (lesions in the upper digestive tract at all dose levels)

Key result

Critical effects observed:

no

Lowest effective dose / conc.:

100 mg/kg bw/day (nominal)

System:

gastrointestinal tract

Organ:

other: upper digestive tract

GROSS PATHOLOGY (PARENTAL ANIMALS) / HISTOPATHOLOGY (PARENTAL ANIMALS)

Treatment of male and female Wistar rats with up to 1000 and 600 mg/kg bw/d of the test substance led to treatment-related findings in the upper digestive tract. The hyperplasia, hydropic degeneration of squamous cells and inflammatory cell infiltrates, occasionally with multinucleated giant cells in the forestomach as well as the hyperemia, eosinophilic cytoplasmic change, increased mitotic figures, submucosal edema and inflammatory cell infiltrates in the glandular stomach were regarded to be signs of a slight irritating effect of the test substance. These findings were almost exclusively observed around or at the margo plicatus and in the glandular stomach in the fundic area. They were regarded to be adverse. The increase in thickness in the duodenum was also regarded to be treatment-related and as the specific mechanism could not be determined, it was also judged to be adverse in nature. All these adverse findings described above are regarded to be a local irritating effect but no systemic toxicity.

Conclusions:

Under the conditions of the study, the NOAEL has been determined as 600 mg/kg bw/day in relation to parental systemic toxicity (male/female); 100 mg/kg bw/day in relation to local effect (female – histopathology (lesions in the upper digestive tract at all dose levels) and < 100 mg/kg bw/day in relation to local effect (male – histopathology (lesions in the upper digestive tract at all dose levels).

Executive summary:

An OECD 422 study has been performed on the substance Bis-Aminopropyl Diglycol (also known as 3,3'-oxybis(ethyleneoxy)bis(propylamine); EC Number 224-207-2; CAS Number 4246-51-9). The study is considered as a guideline study conducted in accordance with GLP.

The Bis-Aminopropyl Diglycol was administered to Wistar- strain rats via gavage at concentrations of 100, 300 and 1000 (reduced to 600 on study day 7) mg/kg bw/day. 

Cage-side observations and detailed clinical observations were recorded. Body weight, food consumption, clinical chemistry including haematology and gross pathology were noted to being treatment-related.

Under the conditions of the study, the NOAEL has been determined as 600 mg/kg bw/day in relation to parental systemic toxicity (male/female); 100 mg/kg bw/day in relation to local effect (female – histopathology (lesions in the upper digestive tract at all dose levels) and < 100 mg/kg bw/day in relation to local effect (male – histopathology (lesions in the upper digestive tract at all dose levels).

Reference 2

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

1946 and earlier

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

test procedure in accordance with national standard methods with acceptable restrictions

Reason / purpose for cross-reference:

read-across source

Qualifier:

no guideline followed

Principles of method if other than guideline:

Two-years feeding study

GLP compliance:

no

Species:

rat

Strain:

Osborne-Mendel

Sex:

male

Details on test animals or test system and environmental conditions:

- individual caging
- temperature- and humidity controlled room
- feed and water ad lib.

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

Ground commercial rat biscuits with 1 % cod-liver oil as basic diet. Maleic acid was mixed with the basic diet by means of a rotary batch mixer.

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

2 years

Frequency of treatment:

daily

Dose / conc.:

0.5 other: nominal in diet

Remarks:

%

Dose / conc.:

1 other: nominal in diet

Remarks:

%

Dose / conc.:

1.5 other: nominal in diet

Remarks:

%

No. of animals per sex per dose:

12 (only males)

Control animals:

yes, plain diet

Positive control:

not appropriate

Observations and examinations performed and frequency:

BODY WEIGHT: Yes
- Time schedule for examinations: once weekly

FOOD CONSUMPTION: yes
- Food consumption for each animal determined once weekly

Sacrifice and pathology:

GROSS PATHOLOGY: Yes

HISTOPATHOLOGY: Yes
routinely: heamatoxylin-eosin stained paraffin sections of lung, heart, liver, spleen, pancreas, stomach, small intestine, adrenal, testes
frequently enough to make it reasonably certain that no changes in them were being caused by the test substance: colon, bone marrow, leg bones, leg muscles, lymph nodes, thyroid, parathyroid.

Clinical signs:

effects observed, treatment-related

Mortality:

mortality observed, treatment-related

Body weight and weight changes:

effects observed, treatment-related

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

not specified

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 specified

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

no effects observed

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Histopathological findings: neoplastic:

no effects observed

Details on results:

CLINICAL SIGNS AND MORTALITY
- increased mortality rate at 18 months at 1 and 1.5 %
- increased mortality rate at 24 months: 12/12 at 1.5 %, 10/12 at 1 %, 10/12 at 0.5 % (controls: 6/12)

BODY WEIGHT AND WEIGHT GAIN
- significantly reduced at concentrations of 1 and 1.5 %

FOOD CONSUMPTION
- no findings

GROSS PATHOLOGY
- no findings

HISTOPATHOLOGY: NON-NEOPLASTIC
- enlarged and irregularly shaped epithelial cells in small to moderate numbers of renal tubules, generally the proximal convoluted, in 4 rats at 1.5 % and in 3 rats at 1 %
- distinctly more atrophy of the liver and less calcification in large arteries at 1.5 %
- more atrophy of the testis at 1.5 %
- inanition may have been one cause for atrophy of liver and testis

HISTOPATHOLOGY: NEOPLASTIC
- no findings

Dose descriptor:

NOAEL

Based on:

test mat.

Sex:

male

Basis for effect level:

other: no NOAEL was identified

Remarks on result:

not determinable

Critical effects observed:

not specified

Conclusions:

Toxic effects were observed at concentrations of 0.5 %, 1 % and 1.5 % of maleic acid in the diet, fed to male rats for two years. The main findings were increased mortality rate, reduced body weight/body weight gain and histopathological changes in liver, testis and kidneys.

Executive summary:

Maleic acid, mixed with the diet at concentrations of 0.5, 1 and 1.5 %, was fed to groups of 12 male rats each for two years. A negative control group (12 male rats) was fed only diet.

 

Body weight and food consumption was determined individually in weekly intervals. Necropsy was performed on all animals as far as possible. Lung, heart, liver, spleen, pancreas, stomach, small intestine, adrenal, testes were routinely subjected to microscopic evaluation, colon, bone marrow, leg bones, leg muscles, lymph nodes, thyroid, parathyroid in some cases.

 

- Body weight and body weight gain were significantly reduced at concentrations of 1 and 1.5 %.

 

- Mortality rates were increased at 18 months at 1 and 1.5 % and at each concentration tested at 24 months: 12/12 at 1.5 %, 10/12 at 1 %, 10/12 at 0.5 % (controls: 6/12).

 

- No major visceral damages were noted in the test substance treated rats.

 

- Rats fed 1.5 % maleic acid showed more atrophy of the liver and of the testis than did the remaining groups. Inanition seemed at least partly responsible for these differences.

 

- Enlarged and irregularly shaped epithelial cells in small to moderate numbers of renal tubules, generally the proximal convoluted, were found in 4 rats at 1.5 % and in 3 rats at 1 % - Distinctly more atrophy of the liver and less calcification in large arteries were noted at 1.5 %.

 

Toxic effects were observed at concentrations of 0.5 %, 1 % and 1.5 % of maleic acid in the diet, fed to male rats for two years. The main findings were increased mortality rate, reduced body weight/body weight gain and histopathological changes in liver, testis and kidneys. No NOAEL was defined.

Reference 3

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

1975

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

Data from handbook or collection of data, read-across from maleic anhydride

Justification for type of information:

Maleic anhydride. However, maleic anhydride hydrolyses under test conditions. As a result, it is believed that maleic acid and its salts were the test materials investigated in this study.

Reason / purpose for cross-reference:

read-across source

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 409 (Repeated Dose 90-Day Oral Toxicity Study in Non-Rodents)

Deviations:

not specified

Principles of method if other than guideline:

Maleic anhydride was administered to dogs in the feed for 90 days. Several observations were made, including hematology, urinalysis, various clinical chemistry and others.

GLP compliance:

no

Specific details on test material used for the study:

Maleic anhydride (>99.5%). However, maleic anhydride hydrolyses under test conditions. As a result, it is believed that maleic acid and its salts were the test materials investigated in this study.

Species:

dog

Strain:

Beagle

Sex:

male/female

Route of administration:

oral: feed

Vehicle:

not specified

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

90 days

Frequency of treatment:

7 days/week, ad libitum

Dose / conc.:

0 mg/kg bw/day (nominal)

Remarks:

in diet

Dose / conc.:

20 mg/kg bw/day (nominal)

Remarks:

in diet

Dose / conc.:

40 mg/kg bw/day (nominal)

Remarks:

in diet

Dose / conc.:

60 mg/kg bw/day (nominal)

Remarks:

in diet

No. of animals per sex per dose:

4

Control animals:

yes, concurrent vehicle

Observations and examinations performed and frequency:

Body weights and food consumption were recorded, and the dogs were observed for signs of toxicity. Clinical parameters were evaluated by determination made prior to the beginning of the study and at intervals throughout the test period. These included hematological studies, urinalysis and various clinical chemistry studies.

Sacrifice and pathology:

Gross examination was conducted at necropsy after the exposure period, organ weights were examined, and tissues were obtained for histopathological evaluation.

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

food intake decreased for the first few weeks in both sexes at 60 mg/kg

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

equivocal decreases in packed cell volume and hemoglobin concentration of male dogs at 60 mg/kg bw after 83 days

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

no effects observed

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

not specified

Histopathological findings: neoplastic:

not specified

Other effects:

not specified

Details on results:

Food intake was decreased for the first few weeks in both sexes of dogs receiving 60 mg/kg/day. Hematological studies conducted after 83 days of exposure indicated equivocal decreases in packed cell volume and hemoglobin concentration of male dogs receiving 60 mg/kg/day. No other changes were observed that were considered relatable to the ingestion of diets containing maleic anhydride.

Dose descriptor:

NOEL

Effect level:

60 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: clinical signs; mortality; body weight; haematology; clinical chemistry

Critical effects observed:

not specified

Conclusions:

The NOEL of a 90-days dietary feeding study with Beagle dogs was 60 mg/kg/day, which was the highest concentration tested.

Executive summary:

Maleic anhydride was mixed with the food and administered on 7 day/week ad lib., for 90 consecutive days to four Beagle dogs/sex/dose. Doses were 0, 20, 40 or 60 mg/kg body weight.

Body weights and food consumption were recorded, and the dogs were observed for signs of toxicity. Clinical parameters were evaluated by determination made prior to the beginning of the study and at intervals throughout the test period. These included hematological studies, urinalysis and various clinical chemistry studies. Gross examination was conducted at necropsy after the exposure period, organ weights were examined, and tissues were obtained for histopathological evaluation.

Food intake was decreased for the first few weeks in both sexes of dogs receiving 60 mg/kg/day. Hematological studies conducted after 83 days of exposure indicated equivocal decreases in packed cell volume and hemoglobin concentration of male dogs receiving 60 mg/kg/day. No other changes were observed that were considered relatable to the ingestion of diets containing maleic anhydride.

The NOEL was defined to be 60 mg/kg/day for both sexes of Beagle dogs.

Reference 4

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

Data from handbook or collection of data, read-across from maleic anhydride

Justification for type of information:

Maleic anhydride. However, maleic anhydride hydrolyses under test conditions. As a result, it is believed that maleic acid and its salts were the test materials investigated in this study.

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

OECD Guideline 452 (Chronic Toxicity Studies)

Deviations:

not specified

Principles of method if other than guideline:

Maleic anhydride was administered to rats in the feed for two years days. Several observations were made, including hematology, urinalysis, various clinical chemistry and others.

GLP compliance:

not specified

Specific details on test material used for the study:

Maleic anhydride. However, maleic anhydride hydrolyses under test conditions. As a result, it is believed that maleic acid and its salts were the test materials investigated in this study.

Species:

rat

Strain:

Fischer 344

Sex:

male/female

Route of administration:

oral: feed

Vehicle:

not specified

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The analysis of the diet for maleic anhydride was conducted 2 to 4 years after completion of the in-life phase of the study. A sample of each diet was removed, refrigerated or frozen for future chemical determination of the concentration of maleic anhydride. The GC-MS measurements were (on average of random samples) 69-75% of the expected for male and female diets, respectively.

Duration of treatment / exposure:

24 months

Frequency of treatment:

7 days/week

Dose / conc.:

0 mg/kg diet

Dose / conc.:

10 mg/kg diet

Dose / conc.:

32 mg/kg diet

Dose / conc.:

100 mg/kg diet

No. of animals per sex per dose:

125

Control animals:

yes, concurrent vehicle

Details on study design:

Scheduled interim termination timepoints were at 6, 12, and 18 months with final study termination at 24 months.

Observations and examinations performed and frequency:

Clinical signs of toxicity, body weights and food consumption were monitored and extensive histopathological examinations were conducted. Additionally, the eyes of all animals were examined by ophthalmoscope and hematology, clinical chemistry and urine parameters were assessed in five animals/sex/dose.

Sacrifice and pathology:

Extensive histopathological examinations were conducted.

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Slightly reduced in males at 32 and 100 mg/kg.

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

Slightly reduced at 32 and 100 mg/kg, for a limited period of time.

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

effects observed, treatment-related

Description (incidence and severity):

higH incidence of cataracts.

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

no effects observed

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Details on results:

There was only marginal toxicity which was evidenced by small (<6%), but dose-related, decrease in body weights of male rats fed 32 and 100 mg/kg/day compared to the controls. The female rats fed 32 and 100 mg/kg/day also had reduced body weights, but the reductions were smaller and of shorter duration than those observed in males. Food consumption was also slightly reduced during limited periods during the study for animals in the mid- and high-dose groups. Neither neurologic nor ophthalmologic evaluations revealed differences between treated and control animals. There was a high incidence of cataracts in the animals of this study, with 100% of the animals examined at 18 month and at study termination bearing cataracts. The severity of these cataracts was independent of maleic anhydride consumption. Hematology, clinical chemistry, gross or histopathological evaluations (including the kidneys) showed no differences between treated and control animals that were considered related to maleic anhydride exposure.

Dose descriptor:

LOEL

Effect level:

32 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: reduced body weights, slightly reduced food consumption

Dose descriptor:

NOEL

Effect level:

10 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: body weights, food consumption

Critical effects observed:

not specified

Conclusions:

The LOEL of a 2-years dietary feeding study with rats was 32 mg/kg/day, the NOEL was 10 mg/kg/day.

Executive summary:

Maleic anhydride was mixed with the food and administered on 7 day/week ad lib., for 2 years to Fischer 344 rats. Doses were 0, 10, 32 or 100 mg/kg body weight. 126 animals were used per group.

 

Scheduled interim termination timepoints were at 6, 12, and 18 months with final study termination at 24 months. Clinical signs of toxicity, body weights and food consumption were monitored and extensive histopathological examinations were conducted. Additionally, the eyes of all animals were examined by ophthalmoscope and hematology, clinical chemistry and urine parameters were assessed in five animals/sex/dose. The analysis of the diet for maleic anhydride was conducted 2 to 4 years after completion of the in-life phase of the study. A sample of each diet was removed, refrigerated or frozen for future chemical determination of the concentration of maleic anhydride. The GC-MS measurements were (on average of random samples) 69-75% of the expected for male and female diets, respectively. There was a problem with the animal room lighting control system which resulted in exposure to continuous light for an unknown period.

 

There was only marginal toxicity which was evidenced by small (<6%), but dose-related, decrease in body weights of male rats fed 32 and 100 mg/kg/day compared to the controls. The female rats fed 32 and 100 mg/kg/day also had reduced body weights, but the reductions were smaller and of shorter duration than those observed in males. Food consumption was also slightly reduced during limited periods during the study for animals in the mid- and high-dose groups. Neither neurologic nor ophthalmologic evaluations revealed differences between treated and control animals. There was a high incidence of cataracts in the animals of this study, with 100% of the animals examined at 18 month and at study termination bearing cataracts. The severity of these cataracts was independent of maleic anhydride consumption. Hematology, clinical chemistry, gross or histopathological evaluations (including the kidneys) showed no differences between treated and control animals that were considered related to maleic anhydride exposure.

 

The LOEL was defined to be 32 mg/kg/day and the NOEL was 10 mg/kg/day for both sexes.

Reference 5

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

1975

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

Data from handbook or collection of data, read-across from maleic anhydride

Justification for type of information:

Data from handbook or collection of data, read-across from maleic anhydrideMaleic anhydride. However, maleic anhydride hydrolyses under test conditions. As a result, it is believed that maleic acid and its salts were the test materials investigated in this study.

Reason / purpose for cross-reference:

read-across source

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)

Deviations:

not specified

Principles of method if other than guideline:

Maleic anhydride was administered to rats in the feed for 90 days. The parameters evaluated included clinical signs (appearance and demeanor), body weight, food consumption, hematological and clinical chemistry evaluations, urinalysis and a urine concentration test, organ weights, gross and histopathology.

GLP compliance:

no

Specific details on test material used for the study:

Maleic anhydride (>99.5%). However, maleic anhydride hydrolyses under test conditions. As a result, it is believed that maleic acid and its salts were the test materials investigated in this study.

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Route of administration:

oral: feed

Vehicle:

not specified

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

90 days

Frequency of treatment:

7 days/week, ad libitum

Dose / conc.:

0 mg/kg bw/day (nominal)

Remarks:

in diet

Dose / conc.:

20 mg/kg bw/day (nominal)

Remarks:

in diet

Dose / conc.:

40 mg/kg bw/day (nominal)

Remarks:

in diet

No. of animals per sex per dose:

no data

Control animals:

yes, concurrent vehicle

Observations and examinations performed and frequency:

The parameters evaluated included clinical signs (appearance and demeanor), body weight, food consumption, hematological and clinical chemistry evaluations, urinalysis and a urine concentration test.

Sacrifice and pathology:

The parameters evaluated included organ weights, gross and histopathology

Clinical signs:

not specified

Mortality:

not specified

Body weight and weight changes:

not specified

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

not specified

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

not specified

Histopathological findings: neoplastic:

not specified

Other effects:

not specified

Details on results:

No data.

Dose descriptor:

NOEL

Effect level:

40 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No data

Critical effects observed:

not specified

Conclusions:

The NOEL of a 90-days dietary feeding study with rats was 40 mg/kg/day, which was the highest concentration tested.

Executive summary:

Maleic anhydride was mixed with the food and administered on 7 day/week ad lib., for 90 consecutive days to Sprague-Dawley rats. Doses were 0, 20 or 40 mg/kg body weight.

The parameters evaluated included clinical signs, body weight, food consumption, haematological and clinical chemistry evaluations, urinalysis, organ weights, gross and histopathology.

The NOEL was defined to be 40 mg/kg/day for both sexes of rats.

Reference 6

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

1975

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

Data from handbook or collection of data, read-across from maleic anhydride

Justification for type of information:

Maleic anhydride. However, maleic anhydride hydrolyses under test conditions. As a result, it is believed that maleic acid and its salts were the test materials investigated in this study.

Reason / purpose for cross-reference:

read-across source

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)

Deviations:

not specified

Principles of method if other than guideline:

Maleic anhydride was administered to rats in the feed for 90 days. Several observations were made, including hematology, urinalysis, various clinical chemistry and others.

GLP compliance:

no

Specific details on test material used for the study:

Maleic anhydride (>99.5%). However, maleic anhydride hydrolyses under test conditions. As a result, it is believed that maleic acid and its salts were the test materials investigated in this study.

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Route of administration:

oral: feed

Vehicle:

not specified

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

90 days

Frequency of treatment:

7 days/week, ad libitum

Dose / conc.:

0 mg/kg bw/day (nominal)

Remarks:

in diet

Dose / conc.:

100 mg/kg bw/day (nominal)

Remarks:

in diet

Dose / conc.:

250 mg/kg bw/day (nominal)

Remarks:

in diet

Dose / conc.:

600 mg/kg bw/day (nominal)

Remarks:

in diet

No. of animals per sex per dose:

no data

Observations and examinations performed and frequency:

The parameters evaluated included clinical signs (appearance and demeanor), body weight, food consumption, hematological and clinical chemistry evaluations, urinalysis and a urine concentration test.

Sacrifice and pathology:

The parameters evaluated included organ weights, gross and histopathology.

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

effects observed, treatment-related

Description (incidence and severity):

slight proteinuria in both sexes at 600 mg/kg/day

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

At 600 mg/kg/day, there was increased relative liver weight in males, increased relative/absolute kidney weights in both sexes (p<0.05). At 250 mg/kg/day, there was increased relative/absolute kidney weights in males (p<0.05).

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

Grossly observed kidney changes were seen in males fed maleic anhydride 100, 250, and 600 mg/kg/day. The changes were characterized by increased size, pale discoloration, and evidence of dilated tubules in the cortex.

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

Grossly observed kidney changes were seen in males fed maleic anhydride 100, 250, and 600 mg/kg/day. The changes were characterized by increased size, pale discoloration, and evidence of dilated tubules in the cortex.

Histopathological findings: neoplastic:

effects observed, treatment-related

Description (incidence and severity):

In the lung of one male at both 250 and 600 mg/kg/day, there was a singular nodular lesion observed grossly that was microscopically diagnosed as a pulmonary adenoma.

Other effects:

not specified

Details on results:

At 600 mg/kg/day, there was slight proteinuria in both sexes, increased relative liver weight in males, increased relative/absolute kidney weights in both sexes (p<0.05). At 250 mg/kg/day, there was increased relative/absolute kidney weights in males (p<0.05). Grossly observed kidney changes were seen in males fed maleic anhydride 100, 250, and 600 mg/kg/day. The changes were characterized by increased size, pale discoloration, and evidence of dilated tubules in the cortex. Microscopically, the kidneys showed varying degrees of nephrosis, being most severe in the high-dose group. These changes consisted of diffuse tubular dilatation, hypertrophy, and degeneration and regeneration of the tubular cells in the cortical portion of the nephron. A decrease in severity of these changes was observed at lower dose levels, with the kidneys from the 100 mg/kg dose group showing minimal laterations. Similar changes were observed in the kidneys of females, but were generally limited to the high-dose group and were much less severe. In the lung of one male at both 250 and 600 mg/kg/day, there was a singular nodular lesion observed grossly that was microscopically diagnosed as a pulmonary adenoma.

Dose descriptor:

LOEL

Effect level:

100 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: grossly observed kidney changes, mainly in males

Critical effects observed:

not specified

Conclusions:

The LOEL of a 90-days dietary feeding study with rats was 100 mg/kg/day, which was the lowest concentration tested.

Executive summary:

Maleic anhydride was mixed with the food and administered on 7 day/week ad lib., for 90 consecutive days to Sprague-Dawley rats. Doses were 0, 100, 250 or 600 mg/kg body weight.

The parameters evaluated included clinical signs (appearance and demeanour), body weight, food consumption, haematological and clinical chemistry evaluations, urinalysis and a urine concentration test, organ weights, gross and histopathology.

At 600 mg/kg/day, there was slight proteinuria in both sexes, increased relative liver weight in males, increased relative/absolute kidney weights in both sexes (p<0.05). At 250 mg/kg/day, there was increased relative/absolute kidney weights in males (p<0.05). Grossly observed kidney changes were seen in males fed maleic anhydride 100, 250, and 600 mg/kg/day. The changes were characterized by increased size, pale discoloration, and evidence of dilated tubules in the cortex. Microscopically, the kidneys showed varying degrees of nephrosis, being most severe in the high-dose group. These changes consisted of diffuse tubular dilatation, hypertrophy, and degeneration and regeneration of the tubular cells in the cortical portion of the nephron. A decrease in severity of these changes was observed at lower dose levels, with the kidneys from the 100 mg/kg dose group showing minimal lacerations. Similar changes were observed in the kidneys of females, but were generally limited to the high-dose group and were much less severe. In the lung of one male at both 250 and 600 mg/kg/day, there was a singular nodular lesion observed grossly that was microscopically diagnosed as a pulmonary adenoma.

The LOEL was defined to be 100 mg/kg/day for both sexes of rats.

Reference 7

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

1977

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

Data from handbook or collection of data, read-across from maleic anhydride

Justification for type of information:

Maleic anhydride. However, maleic anhydride hydrolyses under test conditions. As a result, it is believed that maleic acid and its salts were the test materials investigated in this study.

Reason / purpose for cross-reference:

read-across source

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 452 (Chronic Toxicity Studies)

Deviations:

not specified

Principles of method if other than guideline:

Maleic anhydride was administered to rats in the feed for 183 days. Several observations were made, including hematology, urinalysis, various clinical chemistry and others.

GLP compliance:

no

Specific details on test material used for the study:

Maleic anhydride (>99.5%). However, maleic anhydride hydrolyses under test conditions. As a result, it is believed that maleic acid and its salts were the test materials investigated in this study.

Species:

rat

Strain:

Sprague-Dawley

Sex:

male

Route of administration:

oral: feed

Vehicle:

not specified

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

90 days

Frequency of treatment:

7 days/week, ad libitum

Dose / conc.:

0 mg/kg bw/day (nominal)

Remarks:

in diet

Dose / conc.:

250 mg/kg bw/day (nominal)

Remarks:

in diet

Dose / conc.:

600 mg/kg bw/day (nominal)

Remarks:

in diet

No. of animals per sex per dose:

50 animals/treatment group, 75 animals/control group

Control animals:

yes, concurrent vehicle

Observations and examinations performed and frequency:

Body weights and food consumption were recorded, and the animals were observed for signs of toxicity. Clinical parameters were evaluated for five rats/dose on day 174 of the study. Urine samples were collected on days 12, 84, and 176 of the study for analysis, as well as a urine concentration test on days 84-85 and 176-177. Clinical chemistry determinations were also made on blood samples taken from 10 rats/dose at the 90-day and 183 necropsies.

Sacrifice and pathology:

Necropsies, organ weights, and histopathology were conducted on rats at the 90-day and 183-day termination timepoints. Organ weights were for kidney and liver (90-day); and kidneys, liver, brain, heart, and testes (183-day). For histopathology, only the lung, kidney, and liver were examined for both termination timepoints.

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

no effects observed

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

liver, kidneys, testes, brain, heart

Gross pathological findings:

not specified

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

kidneys: degenerative, hypertrophic, and regenerative, dose-related changes

Histopathological findings: neoplastic:

no effects observed

Other effects:

not specified

Details on results:

No statistically significant changes in mean body weights and food consumption between treated and control animals. Neither was there any meaningful (or significant) differences in haematology, urinalysis, urine concentration, or clinical chemistry tests. At the 90-day timepoint, the relative liver weight for the rats in the high-dose level and the absolute/relative kidney weights for both the 250 and 600 mg/kg groups were significantly higher compared to the controls (p<0.05). At 183 days, there was a significant increase in the absolute and relative liver and kidney weights for both treatment groups, increased relative testes weight, brain and heart, and decreased fasted body weights for rats at 600 mg/kg/day (p<0.05). Treatment-related changes were present in the kidneys of rats terminated at 90 and 183 days. The observed changes indicated a marked accentuation of the spontaneously occurring findings seen in the control animals. The changes in the controls and treated animals included: individual tubules that were dilated and contained eosinophilic staining casts, granular degeneration of the epithelial cells lining these tubules, tubular collapse and atrophy with peritubular fibrosis, focal mononuclear inflammatory cell infiltrates, glomeruli that showed thickening of the basement membrane, thickening and epithelialization of Bowmans Capsule, and occasionally showed either focal or diffuse sclerosis of the glomerular tufts. Both tubular and glomerular changes in treated animals were more severe than controls, and much more severe in the treated rats at 183 days compared to 90 days. In addition to the focal nature of the lesion (in controls), the tubules throughout the cortex of treated rats showed a generalized dilatation and hypertrophy. There were more degenerative tubules and tubules showing mitotic activity in treated versus controls. The degree of degenerative, hypertrophic, and regenerative changes was dose-related. In many of the 600 mg/kg dose group animals, there was a marked decrease in the amount of functional tissue in the kidney. Livers of maleic anhydride-treated rats at 183 days showed changes characterized by swollen individual hepatocytes having vacuolated cytoplasm.

Dose descriptor:

LOEL

Effect level:

250 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male

Basis for effect level:

other: organ weights (liver, kidneys, testes, brain, heart; degenerative, hypertrophic and regenerative kidney changes

Critical effects observed:

not specified

Conclusions:

The LOEL of a 183-days dietary feeding study with rats was 250 mg/kg/day, which was the lowest concentration tested.

Executive summary:

Maleic anhydride was mixed with the food and administered on 7 day/week ad lib., for 183 consecutive days to male Sprague-Dawley rats. Doses were 0, 250 or 600 mg/kg body weight. 50 animals were used for each treatment group and 75 for the control group.

Body weights and food consumption were recorded, and the animals were observed for signs of toxicity. Clinical parameters were evaluated for five rats/dose on day 174 of the study. Urine samples were collected on days 12, 84, and 176 of the study for analysis, as well as a urine concentration test on days 84-85 and 176-177. Clinical chemistry determinations were also made on blood samples taken from 10 rats/dose at the 90-day and 183 necropsies. Necropsies, organ weights, and histopathology were conducted on rats at the 90-day and 183-day termination timepoints. Organ weights were for kidney and liver (90-day); and kidneys, liver, brain, heart, and testes (183-day). For histopathology, only the lung, kidney, and liver were examined for both termination timepoints.

No statistically significant changes in mean body weights and food consumption between treated and control animals. Neither was there any meaningful (or significant) differences in haematology, urinalysis, urine concentration, or clinical chemistry tests. At the 90-day timepoint, the relative liver weight for the rats in the high-dose level and the absolute/relative kidney weights for both the 250 and 600 mg/kg groups were significantly higher compared to the controls (p<0.05). At 183 days, there was a significant increase in the absolute and relative liver and kidney weights for both treatment groups, increased relative testes weight, brain and heart, and decreased fasted body weights for rats at 600 mg/kg/day (p<0.05). Treatment-related changes were present in the kidneys of rats terminated at 90 and 183 days. The observed changes indicated a marked accentuation of the spontaneously occurring findings seen in the control animals. The changes in the controls and treated animals included: individual tubules that were dilated and contained eosinophilic staining casts, granular degeneration of the epithelial cells lining these tubules, tubular collapse and atrophy with peritubular fibrosis, focal mononuclear inflammatory cell infiltrates, glomeruli that showed thickening of the basement membrane, thickening and epithelialization of Bowmans Capsule, and occasionally showed either focal or diffuse sclerosis of the glomerular tufts. Both tubular and glomerular changes in treated animals were more severe than controls, and much more severe in the treated rats at 183 days compared to 90 days. In addition to the focal nature of the lesion (in controls), the tubules throughout the cortex of treated rats showed a generalized dilatation and hypertrophy. There were more degenerative tubules and tubules showing mitotic activity in treated versus controls. The degree of degenerative, hypertrophic, and regenerative changes was dose-related. In many of the 600 mg/kg dose group animals, there was a marked decrease in the amount of functional tissue in the kidney. Livers of maleic anhydride-treated rats at 183 days showed changes characterized by swollen individual hepatocytes having vacuolated cytoplasm.

The LOEL was defined to be 250 mg/kg/day for male rats.

Reference 8

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

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

Adequacy of study:

other information

Justification for type of information:

The read-across prediction was based upon the data available on the source substances. The two source substances Maleic Acid and 3,3'-oxybis(ethyleneoxy)bis(propylamine 4,7,10-Trioxatridecan-1,13-diam are mixed together in water in a 2:1 ratio to form the target substance. The only differences are the charged species and that the target substance only exists in an aqueous solution. The source and target substances are all mono-constituent substances. The remaining source substance, Maleic Anhydride was used because this is the substance that can be found in the disseminated ECHA dossier for Maleic Acid, and that Maleic Anhydride hydrolyses under test conditions. As a result, it is believed that Maleic Acid and its salts were the test materials investigated in the studies.

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

NOAEL

100 mg/kg bw/day

Study duration:

subchronic

Species:

rat

System:

gastrointestinal tract

Organ:

duodenum

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

Read across values from the most suitable substances have been used for classification.

The substance does not meet the criteria for classification for single target organ toxicity (repeated exposure) under CLP.