Registration Dossier

Toxicological information

Toxicity to reproduction

Currently viewing:

Administrative data

Endpoint:
toxicity to reproduction
Remarks:
other: OECD 408 with additional histopathological assessment of the reproductive organs
Type of information:
experimental study
Adequacy of study:
key study
Study period:
01 Jul 2014 to 20 Apr 2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was performed according to an established protocol and consistent with Good Laboratory Practice.
Cross-reference
Reason / purpose:
reference to same study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015
Report Date:
2015

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
other: OECD 408 with additional histopathological assessment of the reproductive organs
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Details on test material:
- Name of test material (as cited in study report): ALCHISOR CAL 145
- Substance type: Product
- Physical state: liquid

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
Test animals: The Han Wistar rat was chosen as the animal model for this study as it is an accepted rodent species for preclinical toxicity testing by regulatory agencies. On 10 Jun 2014, 52 male and 52 female Han Wistar rats were received from Charles River UK, Margate, Kent. At the initiation of dosing the animals were 8-9 weeks old and weighed between 212-300 g (males) and 147-194 g (females). The total number of animals used in this study was considered to be the minimum required to properly characterise the effects of the test item. This study was designed such that it did not require an unnecessary number of animals to accomplish its objectives.Environmental conditions: Temperatures of 18-24°C with a relative humidity of 40-75% were maintained. The target temperature and humidity ranges were 19-23oC and 40-70% respectively. A 12 hour light/12 hour dark cycle and ten or greater air changes per hour with 100% fresh air (no air recirculation) were maintained in the animal rooms.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
other: 0.5% HPMC (Methocel E4M), 0.1% Tween 80 in Milli-Q water
Details on exposure:
The test and control items were administered to the appropriate animals by once daily oral gavage for 90 days. The dose volume for each animal was based on the most recent body weight measurement. The doses were given using a syringe with attached gavage cannula. The first day of dosing was designated as Day 1.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Test item dosing formulations were prepared, based on a method established at the Test Facility (under Study Numbers 432356 and 431130), at appropriate concentrations to meet dosage level requirements. The required amount of test item was weighed and transferred to a pre-labelled container. The appropriate amount of vehicle (control item) was weighed and added to the container and the solution was stirred magnetically until a visibly homogeneous formulation was obtained.The dosing formulations were prepared weekly stored in a refrigerator set to maintain 2-8C, and dispensed daily. The dosing formulations were stirred continuously during dosing. Any residual volumes were discarded. Details of the preparation and dispensing of the test item have been retained in the Study Records.Dose formulation samples were collected for analysis during Week 1, 6 and 12, and were submitted for analysis within 1 week of preparation. All samples to be analysed were transferred in ambient conditions to the analytical laboratory at the Test Facility. Analyses were performed using a validated analytical procedure (AP No. 432356). Duplicate top, middle and bottom (middle only for Group 1, concentration only) samples(1 mL) for each sampling time point were sent to the analytical laboratory at the Test Facility. Triplicate top, middle and bottom samples were retained as backup samples. Concentration results were considered acceptable if mean sample concentration results were within or equal to ± 10% of theoretical concentration. Each individual sample concentration result within or equal to ± 20%. For homogeneity, the criteria for acceptability was a relative standard deviation (RSD) of concentrations of <= 10% for each group.
Duration of treatment / exposure:
90 days
Frequency of treatment:
Once daily oral gavage
Doses / concentrationsopen allclose all
Remarks:
Doses / Concentrations:
0 mg/mL
Basis:
nominal in water
Remarks:
Doses / Concentrations:
10 mg/mL
Basis:
nominal in water
Remarks:
Doses / Concentrations:
30 mg/mL
Basis:
nominal in water
Remarks:
Doses / Concentrations:
100 mg/mL
Basis:
nominal in water
No. of animals per sex per dose:
10 animals per sex per dose group
Control animals:
yes, concurrent vehicle
Details on study design:
The objective of this study was to determine the potential toxicity of Alkenes, C13-C14, hydroformylation products, distn. residues (CAS/No. 90622-29-0) when given by oral gavage for at least 90 days to rats and to evaluate the potential reversibility of any findings after 28 days.Alkenes, C13-C14, hydroformylation products, distn residues (CAS/No. 90622-29-0) was administered using 0.5% HPMC (Methocel E4M), 0.1% Tween 80 in Milli-Q water as the vehicle at a constant dose volume of 10 mL/kg. Dose levels were 0 (control), 20, 30, and 100 mg/mL (corresponding to dosage levels of 0, 100, 300, and 1000 mg/kg/day, respectively). Five males and five females at each of the control and high-dose levels, a total of 20 animals, were designated as recovery animals. Additional histopathological examination of reproductive organs and related tissues was performed in order to address the reproductive toxicity endpoint.

Examinations

Parental animals: Observations and examinations:
All animals were checked twice daily, once in the morning and once in the afternoon throughout the study for general health/mortality and moribundity. All animals were removed from their cages for examination once a week from Week -2 and on the first day of scheduled necropsy. All animals were observed regularly throughout the day on dosing days, with particular attention paid during and for the first hour after dosing and at least once daily on non-dosing days during the dosing period for reaction to treatment. In addition animals were observed at least once daily during the recovery period. The onset, intensity and duration of any signs were recorded. Body weights were recorded for each animal twice during pretreatment, daily during the dosing period and twice weekly during the recovery period.Animals were individually weighed. A weight was recorded on the first day of scheduled necropsy.Food consumption was quantitatively measured twice during pretreatment and as follows during the dosing period: Weekly in Weeks 1 to 12 and over 6 days in Week 13 for main study animals: the first 5 animals per sex in Group 1 and all animals in Groups 2 and 3; Weekly in Weeks 1-13 and over 3 days in Week 14 for main study animals: the last 5 animals per sex in Group 1, all animals in Group 3 and all recovery study animals. Food consumption was measured weekly during the recovery period.Water consumption was monitored throughout the study by visual inspection of the water bottles.All animals were subject to an ophthalmic examination once during pretreatment, Control and High Dose animals were examined in Week 13, and all recovery animals were examined during Week 16 and Week 18.Detailed functional observations (cageside observations, observations in a standardized arena; functional tests; grip strength; pain perception; landing food splay; and motor activity) were performed for all animals once during pretreatment period, once during Week 12 of the dosing period and once for all recovery animals during Week 18.
Postmortem examinations (parental animals):
All animals were weighed, and euthanised by exposure to a rising concentration of carbon dioxide, followed by exsanguination. The animals were euthanised rotating across dose groups such that similar numbers of animals from each group, including controls were necropsied at similar times throughout the day. Animals were not fasted before their scheduled necropsy.All main study and recovery animals were subjected to a complete necropsy examination, which included evaluation of the carcass and musculoskeletal system; all external surfaces and orifices; cranial cavity and external surfaces of the brain; and thoracic, abdominal, and pelvic cavities with their associated organs and tissues. Scheduled necropsy examinations were conducted under the supervision of a veterinarian with appropriate training and experience in animal anatomy and gross pathology. See the following summary for additional pathology measures.

Results and discussion

Results: P0 (first parental animals)

General toxicity (P0)

Clinical signs:
no effects observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
Liver weight was increased in males at 300 and 1000 mg/kg/day and females at 1000 mg/kg/day at end of treatment, with no correlating gross or microscopic findings. After a 28 day recovery period, the increase was resolved, indicating reversibility.
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Other effects:
no effects observed

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
not examined
Reproductive function: sperm measures:
not specified
Reproductive performance:
not specified

Details on results (P0)

No test item-related macroscopic or microscopic findings, including male and female reproductive tissues, were noted among either main study or recovery animals.
Administration of Alkenes, C13-C14, hydroformylation products, distn. residues (CAS/No. 90622-29-0) by once daily oral gavage was well tolerated in rats at levels of 100, 300 and 1000 mg/kg/day. Based on the results, due to reversible changes in the liver weights in males and females the no-observed-adverse-effect level (NOAEL) was considered to be 1000 mg/kg/day.

Effect levels (P0)

Key result
Dose descriptor:
NOEL
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: see 'Remark'

Results: P1 (second parental generation)

General toxicity (P1)

Clinical signs:
no effects observed

Effect levels (P1)

Key result
Remarks on result:
not measured/tested

Results: F1 generation

General toxicity (F1)

Clinical signs:
not examined
Mortality / viability:
not specified
Body weight and weight changes:
not specified
Sexual maturation:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
not specified
Histopathological findings:
not specified

Effect levels (F1)

Key result
Remarks on result:
not measured/tested

Results: F2 generation

General toxicity (F2)

Clinical signs:
not examined

Overall reproductive toxicity

Key result
Reproductive effects observed:
not specified

Any other information on results incl. tables

RESULTS Histopathology of Reproductive Organs and Tissues Terminal and Recovery Euthanasia (Day 91/95/96 and 123) Oral (gavage) administration of Alkenes, C13-C14, hydroformylation products, distn. residues (CAS/No. 90622-29-0) once a day to rats for up to 90 days at doses of 0, 100, 300, or 1000 mg/kg/day resulted in no early decedents by the end of the dosing period. In support of the addressing questions of the potential for reproductive toxicity, gross pathology and histopathology results are summarized in the tables below for reproductive organs and tissues of select test animals. At the end of the treatment period, no test article-related gross or microscopic findings were noted. The microscopic findings observed were considered incidental, of the nature commonly observed in this strain and age of rat, and/or were of similar incidence and severity in control and treated animals and, therefore, were considered unrelated to administration of Alkenes, C13-C14, hydroformylation products, distn. residues.

Summary of Gross Pathology Findings (Day 91/95/96)

      Removal Reason: TERMINAL EUTHANASIA    Male Female
 0  100  300  1000 0 100   300  1000
mg/kg/day   mg/kg/day    mg/kg/day  mg/kg/day   mg/kg/day  mg/kg/day   mg/kg/day  mg/kg/day 
Group 1   Group 2  Group 3  Group 4  Group 1  Group 2  Group 3  Group 4
Number of animals:  10  10  10  10  10  10  10  10
 UTERUS                
 - Submitted  -  -  -  -  10  1  2  10
 - No Visible Lesions  -  -  -  -  0  0  10
 - Dilatation; clear  -  -  -  -  1  2  0

Summary of Gross Pathology Findings (Day 123)

          Removal Reason: RECOVERY EUTHANASIA Male Female
 0  1000  0  1000
 mg/kg/day  mg/kg/day  mg/kg/day  mg/kg/day
 Group 1  Group 4  Group 1  Group 4
 Number of animals:  5  5  5  5
 UTERUS        
 - Submitted  -  -  5  5
 - No Visible Lesions  -  -  3  3
 - Dilatation; clear  -  -  2  0
 - Dilatation  -  -  0  1
 - Cyst  -  -  0  1

Summary of Histopathology Findings (Day 91/95/96)

Removal Reason: TERMINAL EUTHANASIA           Male         Female    
 0  100  300  1000  0  100  300  1000
mg/kg/day  mg/kg/day mg/kg/day  mg/kg/day  mg/kg/day  mg/kg/day  mg/kg/day  mg/kg/day
 Group 1  Group 2  Group 3  Group 4  Group 1  Group 2  Group 3  Group 4
 Number of animals:  10  10  10  10  10  10  10  10
 CERVIX                
 -Examined  -  -  -  -  10  0  0  10
 - No Visible Lesions  -  -  -  -  10  -  -  10
 EPIDIDYMIS                
 - Examined  10  0  0  10  -  -  - -
 - No Visible Lesions  10  -  -  10  -  -  -  -
 GLAND, MAMMARY                
 - Examined  10  0  0  10  10  0  0  10
 - No Visible Lesions  10  -  -  10  10  -  -  10
 GLAND, PROSTATE                
 - Examined  10  0  0  10  -  -  -  -
 - No Visible Lesions  8  -  -  10  -  -  -  -
 - Infiltration, mononuclear cell  2  -  -  0  -  -  -  -
 - ....mild  2  -  -  0  -  -  -  -
 GLAND, SEMINAL VESICLE                
 - Examined  10  0  0  10  -  -  -  -
 - No Visible Lesions  10  -  -  10  -  -  -  -
 OVARY                
 - Examined  -  -  -  -  10  0  0  10
 - No Visible Lesions  -  -  -  -  10  -  -  10
 OVIDUCT                
 - Examined  -  -  -  -  10  0  0  10
 - No Visible Lesions  -  -  -  -  10  -  -  10
 TESTIS                
 - Examined  10  0  0  10  -  -  -  -
 - No Visible Lesions  10  -  -  10  -  -  -  -
 UTERUS                
 - Examined  -  -  -  -  10  1  2  10
 - No Visible Lesions  -  -  -  -  10  1  2  10
 VAGINA                
 - Examined  -  -  -  -  9  0  0  10
 - No Visible Lesions  -  -  -  -  9  -  -  10
 - Not Examined: Not Present in Section  -  -  -  -  1  0  0  0

Summary of Histopathology Findings (Day 123)

Removal Reason: RECOVERY EUTHANASIA             Male   Female
 0  1000  0  1000
 mg/kg/day  mg/kg/day  mg/kg/day  mg/kg/day
Group 1   Group 4  Group 1  Group 4
 Number of animals:  5  5  5  5
 CERVIX        
 - Examined  -  -  5  4
 - No Visible Lesions  -  -  5  4
 - Not Prsent in Wet Tissues  -  -  0  1
 EPIDIDYMIS        
 - Examined  5  5  -  -
 - No Visible Lesions  5  5  -  -
 GLAND, MAMMARY        
 - Examined  5  5  5  5
 - No Visible Lesions  5  5  5  5
 GLAND, PROSTATE        
 - Examined  5  5  -  -
 - No Visible Lesions  5  5  -  -
 GLAND, SEMINAL VESICLE        
 - Examined  5  5  -  -
 - No Visible Lesions  5  5  -  -
 OVARY        
 - Examined  -  -  5  5
 - No Visible Lesions  -  -  5  5
 OVIDUCT        
 - Examined  -  -  5  5
 - No Visible Lesions  -  -  5  5
 TESTIS        
 - Examined  5  5  -  -
 - No Visible Lesions  5  5  -  -
 UTERUS        
 - Examined  -  -  5  5
 - No Visible Lesions  -  -  5  5
 VAGINA        
 - Examined  -  5  5
 - No Visible Lesions  -  -  5  5

Applicant's summary and conclusion

Conclusions:
Regarding potential effects of the registered substance on reproductive tissues, no test article-related gross or microscopic findings were noted. The microscopic findings observed were considered incidental, of the nature commonly observed in this strain and age of rat, and/or were of similar incidence and severity in control and treated animals and, therefore, were considered unrelated to administration of Alkenes, C13-C14, hydroformylation products, distn. residues. Thus the NOEL for effects on reproductive tissues is considered to be 1000 mg/kg/day.
Executive summary:

Oral (gavage) administration of Alkenes, C13-C14, hydroformylation products, distn. residues (CAS/No. 90622-29-0) once a day to rats for up to 90 days at doses of 0, 100, 300, or 1000 mg/kg/day resulted in no early decedents by the end of the dosing period. In support of the addressing questions of the potential for reproductive toxicity, gross pathology and histopathology results are summarized in the tables below for reproductive organs and tissues of select test animals. At the end of the treatment period, no test article-related gross or microscopic findings were noted. The microscopic findings observed were considered incidental, of the nature commonly observed in this strain and age of rat, and/or were of similar incidence and severity in control and treated animals and, therefore, were considered unrelated to administration of Alkenes, C13-C14, hydroformylation products, distn. residues. Thus the NOEL for effects on reproductive tissues is considered to be 1000 mg/kg/day.