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Toxicological information

Toxicity to reproduction

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Administrative data

Endpoint:
screening for reproductive / developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
14 September - 30 November 2009
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
The study was conducted in accordance with international guidelines and in accordance with GLP. All relevant validity criteria were met.
Justification for type of information:
REPORTING FORMAT FOR THE ANALOGUE APPROACH
Refer to Section 13.2 for read-across justification document.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report date:
2010

Materials and methods

Test guidelineopen allclose all
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 OPPTS 870.3650, Combined Repeated Dose Toxicity Study with the Reproduction/ Developmental Toxicity Screening Test, July 2000
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Limit test:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
Undecanal
EC Number:
203-972-6
EC Name:
Undecanal
Cas Number:
112-44-7
Molecular formula:
C11H22O
IUPAC Name:
Undecanal
Test material form:
liquid
Specific details on test material used for the study:
RADIOLABELLING INFORMATION (if applicable)
- Radiochemical purity:
n/a
- Specific activity:
n/a
- Locations of the label:
n/a
- Expiration date of radiochemical substance:
n/a

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material:
Dark, under nitrogen
- Stability under test conditions:
Stable
- Solubility and stability of the test substance in the solvent/vehicle:
Test item formulated in propylene glycol. Analysis conducted to confirm stability.
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium:
n/a

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing:
Test item formulated in propylene glycol.
- Preliminary purification step (if any):
n/a
- Final dilution of a dissolved solid, stock liquid or gel:
n/a
- Final preparation of a solid:
n/a

FORM AS APPLIED IN THE TEST (if different from that of starting material)
n/a

TYPE OF BIOCIDE/PESTICIDE FORMULATION (if applicable)
n/a

OTHER SPECIFICS: n/a

Test animals

Species:
rat
Strain:
other: Crl:WI(Han)
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source:
Charles River Deutschland, Sulzfeld, Germany
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation:
ca. 12 weeks
- Weight at study initiation:
all an im als within ± 20% of the sex mean.
- Fasting period before study:
no
- Housing:
Pre-mating: Animals were housed in groups of 5 animals/sexlcage in Macrolon cages (MIV type, height 18 cm). Mating: Females were caged together with males on a one-to-one-basis in Macrolon cages (MIII type, height 18 cm). Post-mating: Males were housed in their home cage (Macrolon cages, MIV type, height 18 cm) with a maximum of 5 animals/cage. Females were individually housed in Macrolon cages (MIII type, height 18 cm). Lactation: Pups were kept with the dam until termination in Macrolon cages (MIII type, height 18 cm). General: Sterilised sawdust as bedding material (Litalabo, S.P.P.S., Argenteuil, France) and paper as cage-enrichment (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ud), Surrey, United Kingdom) were supplied. Certificates of analysis were examined and then retained in the NOTOX archives. During activity monitoring, animals were housed individually in Macrolon cages (MIII type; height 15 cm) with sterilised sawdust as bedding material. No cage-enrichment was provided during activity monitoring.
- Diet (e.g. ad libitum):
Free access to pelleted rode nt diet (SM R1M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany). Results of analyses for nutrients and contaminants were examined and archived. A sampie (approximately 500 grams) of each batch was retained at room temperature until finalization of the study report.
- Water (e.g. ad libitum):
Free access to tap-water. Certificates of analysis (performed quarterly) were examined and archived.
- Acclimation period:
≥ 5 days

DETAILS OF FOOD AND WATER QUALITY:
Not reported

ENVIRONMENTAL CONDITIONS
- Temperature (°C):
21 ± 3 ºC
- Humidity (%):
40-70 %
- Air changes (per hr):
ca. 15
- Photoperiod (hrs dark / hrs light):
12:12 light:dark

IN-LIFE DATES: From: 14 September 2009 To: 30 November 2009

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
propylene glycol
Details on exposure:
- PREPARATION OF DOSING SOLUTIONS:
Not reported

- DIET PREPARATION
- Rate of preparation of diet (frequency):
n/a
- Mixing appropriate amounts with (Type of food):
n/a
- Storage temperature of food:
n/a

- VEHICLE
- Justification for use and choice of vehicle (if other than water):
Based on trial formulations performed at NOTOX.
- Concentration in vehicle:
Not reported
- Amount of vehicle (if gavage):
Dose volume was 5 mL/kg bw
- Lot/batch no. (if required):
Not reported
- Purity: Not reported
Details on mating procedure:
- M/F ratio per cage: Females were caged together with males on a one-to-one-basis.
- Length of cohabitation: A maximum of 13 days was allowed for mating.
- Proof of pregnancy: Detection of mating was confirmed by evidence of sperm in the vaginal lavage, by staging of the oestrus cycle and/or or by the appearance of an intravaginal copulatory plug. This day was designated Day 0 post-coitum.
- After successful mating each pregnant female was caged (how): Females were individually housed in Macrolon cages (MIII type, height 18 cm). Pups were kept with the dam until termination.
- Any other deviations from standard protocol: No
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Analyses were conducted on a single occasion during the treatment phase, according to a validated method (NOTOX project 492005). Sampies of formulations were analyzed for homogeneity (highest and lowest concentration) and accuracy of preparation (all concentrations). Stability in vehicle over 6 hours at room temperature was also determined (highest and lowest concentration).

The accuracy of preparation was considered acceptable if the mean measured concentrations were 90-110% of the target concentration. Homogeneity was demonstrated if the coefficient of variation was ≤ 10%. Formulations were considered stable if the relative difference before and after storage was maximally 10%.
Duration of treatment / exposure:
Males were exposed for 28 days, Ie. 2 weeks prior to mating, during mating, and up to termination. Females were exposed for at 42-48 days, Ie. during 2 weeks prior to mating, during mating, during post-coitum, and during at least 4 days of lactation. Female nos. 69 and 70 (Group 3) were not dosed during littering.
Frequency of treatment:
Once daily for 7 days per week, approximately the same time each day with a maximum of 6 hours difference between the earliest and latest dose. Animals were dosed up to the day prior to scheduled necropsy.
Doses / concentrationsopen allclose all
Dose / conc.:
100 mg/kg bw/day (nominal)
Dose / conc.:
300 mg/kg bw/day (nominal)
Dose / conc.:
1 000 mg/kg bw/day (nominal)
No. of animals per sex per dose:
40 males and 40 females
Control animals:
yes
Details on study design:
- Dose selection rationale: Based on the results of a 14 day preliminary test conducted at 300 and 500 mg/kg
- Rationale for animal assignment (if not random): Random
- Rationale for selecting satellite groups: n/a
- Post-exposure recovery period in satellite groups: n/a
- Section schedule rationale (if not random): n/a

Examinations

Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Twice daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Daily

BODY WEIGHT: Yes
- Time schedule for examinations: Males and females were weighed on the first day of exposure and weekly thereafter. Mated females were weighed on Days 0, 4, 7, 11, 14, 17 and 20 post-coitum and during lactation on Days 1 and 4.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
- Time schedule for examinations: Subjective appraisal

OTHER: haematology (F0) / clinical chemistry (F0) / gross pathology (F0) / nerurobehavioural examination (F0)
Sperm parameters (parental animals):
Of the selected 5 males of the control and high dose group, additional slides of the testes were prepared to examine staging of spermatogenesis. The testes were processed, sectioned at 3-4 micrometers, and stained with PAS/haematoxylin (Klinipath, Duiven, The Netherlands).
Litter observations:
STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: no
- If yes, maximum of [...] pups/litter ([...]/sex/litter as nearly as possible); excess pups were killed and discarded.

PARAMETERS EXAMINED
The following parameters were examined in F1 offspring:
Gestation index, duration of gestation, number of dead and Iiving pups at first Iitter check, sex ratio, postnatal loss, viability index, and early postnatal pup development (mortality, clinical signs, body weight and external macroscopy)

GROSS EXAMINATION OF DEAD PUPS:
external

ASSESSMENT OF DEVELOPMENTAL NEUROTOXICITY: no

ASSESSMENT OF DEVELOPMENTAL IMMUNOTOXICITY: no
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: All surviving animals following completion of mating
- Maternal animals: All surviving animals (day 5-6 lactation of F1)

GROSS NECROPSY
- Gross necropsy consisted of;
Cervix
Prostate gland
Clitoral gland
Seminal vesicles
Coagulation gland
Testes
Epididymides
Uterus
Ovaries
Vagina
Preputial gland
All gross lesions

HISTOPATHOLOGY / ORGAN WEIGHTS
The tissues indicated in Table [2] were prepared for microscopic examination and weighed, respectively.
Postmortem examinations (offspring):
SACRIFICE
- The F1 offspring were sacrificed at 5-6 days of age.
- These animals were subjected to postmortem examinations (macroscopic examination) as follows: All pups were sexed and descriptions of all external abnormalities were recorded. The stomach was examined for the presence of milk. If possible, defects or cause of death were evaluated.
Statistics:
The following statistical methods were used to analyse the data:
- If the variables could be assumed to follow a normal distribution, the Dunnett-test (Dunnett, 1955) (many-to-one t-test) based on a pooled variance estimate was applied for the comparison of the treated groups and the control groups for each sex.
- The Steel-test (Miller, 1981) (many-to-one rank test) was applied if the data could not be assumed to follow a normal distribution.
- The Fisher Exact-test (Fisher, 1950) was applied to frequency data.

All tests were two-sided and in all casesp < 0.05 was accepted as the lowest level of significance.

Group means were calculated for continuous data and medians were calculated for discrete data (scores) in the summary tables. Test statistics were calculated on the basis of exact values for means and pooled variances. Individual values, means and standard deviations may have been rounded off before printing. Therefore, two groups may display the same printed means for a given parameter, yet display different test statistics values.

No statistical analysis was performed on histopathology findings.

The number of implantation sites was subjected to the Kruskal-Wallis nonparametrie ANOVA test (Kruskal, 1952) to determine intergroup difference. If the results of the ANOVA were significant (p<0.05), the Wilcoxon test (Wilcoxon, 1945) was applied to the data to compare the treated groups to the control group.
Reproductive indices:
For each group the following calculations were performed;
% mating females
Fertility index females
Conception rate
Gestation index
Duration of gestation
Offspring viability indices:
For each group the following calculations were performed;
% live males
% live females
% post-natal loss (Day 0-4)
Viability index

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
No clinical signs of toxicity were noted during the observation period.

Salivation seen during/after dosing among all animals of the 300 and 1000 mg/kg dose group during most of the treatment period was considered to be a physiological response rather than a sign of systemic toxicity considering the nature and minor severity of the effect and its time of occurrence (Le. after dosing). This sign may be related to irritancy/taste of the test substance. Rales, scabs and alopecia occurred within the range of background findings to be expected for rats of this age and strain which are housed and treated under the conditions in this study. At the incidence observed, these were considered signs of no toxicological significance.
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Description (incidence and severity):
Body weights and body weight gain of treated animals remained in the same range as controls over the treatment period.
Food consumption and compound intake (if feeding study):
effects observed, non-treatment-related
Description (incidence and severity):
No toxicologically significant changes in food consumption before or after allowance for body weight were noted.

The statistically significant lower food consumption of females at 1000 mg/kg between Days 0-4 of the post-coitum phase was of a temporary and slight nature, and therefore considered to be of no toxicological relevance.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not examined
Haematological findings:
no effects observed
Description (incidence and severity):
Haematological parameters of treated rats were considered not to have been affected by treatment.
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
A statistically significant higher potassium value was measured for males at 1000 mg/kg/day, but the mean remained within the range considered normal for rats of this age and strain. In females at 1000 mg/kg/day, a higher mean cholesterollevel was recorded (being slightly outside the range considered normal for rats of this age and strain) along with notably higher bile acid levels in some females (means not statistically significant). Given that these changes were generally slight in nature, were not present as a group response (bile acids) and occurred in the absence of supportive morphological changes, these were considered to be of no toxicological relevance.

Other statistically significant variations in clinical biochemistry parameters were considered to be of no toxicological relevance as these occurred in the absence of a treatment-related distribution and/or remained within the range considered normal for rats of this age and strain. These variations consisted of higher or lower alkaline phosphatase activity (ALP) in males and females at 300 mg/kg/day, lower total bilirubin level in males at 300 mg/kg/day, higher urea level in males at 300 mg/kg/day, higher cholesterollevel in males at 300 and 1000 mg/kg/day, and lower urea level in females at 1000 mg/kg/day.
Urinalysis findings:
not examined
Behaviour (functional findings):
no effects observed
Description (incidence and severity):
Hearing ability, pupillary reflex, static righting reflex and grip strength were normal in all animals.

The variation in motor activity did not indicate a relation with treatment.
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Description (incidence and severity):
Hearing ability, pupillary reflex, static righting reflex and grip strength were normal in all animals.

The variation in motor activity did not indicate a relation with treatment.
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Histopathological changes were confined to the stomach, and consisted of Iymphogranulocytic inflammation and hyperplasia of the squamous epithelium of the forestomach in both sexes at 100, 300 and 1000 mg/kg/day, and ulcer formation in the forestomach in three males at 300 mg/kg/day (correlating to red/black foci in the forestomach of two males) and in one male at 1000 mg/kg/day. Hyperplasia ofthe squamous epithelium was the microscopic correlate to the irregular surface and yellowish discolouration of the forestomach recorded at necropsy at 300 and 1000 mg/kg/day. Congestion of the glandular stomach (correlating to red discolouration of of the glandular mucosa) was observed in two males at 1000 mg/kg/day. There was no microscopic correlate to thickening of the glandular mucosa recorded in three females at 1000 mg/kg/day.
Histopathological findings: neoplastic:
not examined
Other effects:
not examined

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
not examined
Reproductive function: sperm measures:
no effects observed
Reproductive performance:
no effects observed

Details on results (P0)

No reproductive/developmental toxicity was observed at any dose level.

There were no morphological findings in the reproductive organs of either sex, which could be attributed to the test substance.

Effect levels (P0)

Key result
Dose descriptor:
NOAEL
Effect level:
>= 1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Remarks on result:
not determinable due to absence of adverse toxic effects

Target system / organ toxicity (P0)

Key result
Critical effects observed:
no

Results: F1 generation

General toxicity (F1)

Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
Incidental clinical symptoms of pups consisted of small size, red spots on the head/nose and a pale appearance. No relationship with treatment was established for these observations and they were considered to be of no toxicological relevance.
Mortality / viability:
mortality observed, non-treatment-related
Description (incidence and severity):
A total of three pups of the control group, and one pup each at 100, 300 and 1000 mg/kg/day were found dead or missing during lactation. No toxicological relevance was ascribed to these dead/missing pups since the mortality incidence did not show a dose-related trend and remained within the range considered normal for pups of this age.
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
Some surviving pups were small in size. The nature and incidence of these findings remained within the range considered normal for pups of this age, and were therefore considered to be unrelated to treatment.
Food consumption and compound intake (if feeding study):
not examined
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Incidental macroscopic findings among pups found dead included autolysis, cannibalism and absence of milk in the stomach. The nature and incidence of these findings remained within the range considered normal for pups of this age, and were therefore considered to be unrelated to treatment.
Histopathological findings:
not examined
Other effects:
not examined

Developmental neurotoxicity (F1)

Behaviour (functional findings):
not examined

Developmental immunotoxicity (F1)

Developmental immunotoxicity:
not examined

Details on results (F1)

No reproductive/developmental toxicity was observed at any dose level.

Effect levels (F1)

Key result
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
>= 1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Remarks on result:
not determinable due to absence of adverse toxic effects

Target system / organ toxicity (F1)

Key result
Critical effects observed:
no

Overall reproductive toxicity

Key result
Reproductive effects observed:
no

Any other information on results incl. tables

Table 3       Mean body weights and body weight gains/ losses (F0)

 

Sex / Day

Bodyweight (g)

Control

100 ppm

300 ppm

1000 ppm

MALES (TOXICITY TEST GROUP)

Pre-mating

Day 1

(total body weight gain)

321

-

318

-

316

-

319

-

Day 8

(total % body weight gain)

344

(7)

340

(7)

338

(7)

338

(7)

Mating

Day 1

(total % body weight gain)

367

(14)

360

(13)

360

(14)

359

(13)

Day 8

(total % body weight gain)

378

(18)

373

(17)

370

(17)

374

(17)

Day 14

(total % body weight gain)

391

(22)

386

(21)

386

(22)

388

(22)

Day 15

(total % body weight gain)

-

-

-

-

FEMALES (TOXICITY TEST GROUP)

Pre-mating

Day 1

(total % body weight gain)

213

-

213

-

214

-

212

-

Day 8

(total % body weight gain)

221

(4)

219

(3)

221

(4)

216

(2)

Mating

Day 1

(total % body weight gain)

225

(6)

225

(6)

227

(6)

222

(5)

Day 8

(total % body weight gain)

237

(13)

-

-

225

(10)

Day 15

(total % body weight gain)

253

(20)

-

-

-

Day 22

(total % body weight gain)

311

(48)

-

-

-

Post coitum

Day 0

(total % body weight gain)

232

-

230

-

231

-

225

-

Day 4

(total % body weight gain)

245

(6)

244

(6)

247

(7)

238

(6)

Day 7

(total % body weight gain)

251

(8)

251

(9)

253

(9)

244

(8)

Day 11

(total % body weight gain)

264

(14)

264

(15)

268

(16)

259

(15)

Day 14

(total % body weight gain)

276

(19)

277

(21)

278

(20)

268

(19)

Day 17

(total % body weight gain)

296

(28)

299

(30)

299

(30)

292

(29)

Day 20

(total % body weight gain)

331

(43)

335

(46)

339

(47)

327

(45)

Lactation

Day 1

(total % body weight gain)

256

-

258

-

256

-

252

-

Day 4

(total % body weight gain)

268

(5)

272

(6)

271

(6)

263

(5)

* statistically significantly different from control values p < 0.05

** statistically significantly different from control values p < 0.01

 

Table 4       Haematology, clinical chemistry, urinalysis and pathology findings (F0)

 

Doses (ppm)

Control

100

300

1000

Control

100

300

1000

male (toxicity group)

female (toxicity group)

Haematology

- Hb (mmol/L)

9.2

9.3

9.0

9.6

8.6

8.7

8.4

8.5

- RBC (1012/L)

8.06

8.23

8.07

8.33

7.27

7.46

7.33

7.29

- Hct (%)

0.412

0.420

0.405

0.426

0.378

0.388

0.376

0.379

- Retic (% RBC)

3.2

2.8

2.5

2.8

4.7

5.7

6.5

5.7

- MCH (fmol)

1.14

1.13

1.12

1.16

1.19

1.16

1.15

1.16

- MCV (fl)

51.2

51.1

50.3

51.1

52.0

52.1

51.3

52.0

- MCHC (mmol/L)

22.26

22.16

22.32

22.59

22.95

22.34

22.31

22.40

- RDW (%)

13.9

16.4

15.1

13.2

15.7

15.6

15.4

14.6

- WBC (109/L)

7.4

6.2

6.8

8.0

5.4

5.5

4.2

4.6

- Neut (% WBC)

15.4

14.9

11.2

12.7

23.5

27.5

27.5

27.3

- Lymph (% WBC)

82.3

82.2

86.5

84.7

73.5

69.6

69.5

70.1

- Mono (% WBC)

1.5

1.7

1.4

1.5

1.9

2.1

1.9

1.7

- Eos (% WBC)

0.6

0.9

0.7

0.9

1.0

0.8

0.9

0.7

- Bas (% WBC)

0.2

0.2

0.2

0.2

0.1

0.1

0.1

0.2

- PT (secs)

18.3

18.3

18.5

17.8

18.7

18.6

17.9

17.4

- PLT (109/L)

841

765

829

932

1072

1230

1228

1194

- APTT (secs)

15.5

15.1

17.6

15.0

17.3

17.5

19.2

16.0

Blood chemistry

- Glucose (µmol/L)

9.94

10.88

11.66

11.32

 

 

 

 

- Tot. Prot. (g/L)

64.9

65.2

64.5

65.9

 

 

 

 

- Albumin (g/L)

33.9

33.7

32.7

33.7

 

 

 

 

- A/G ratio

 

 

 

 

 

 

 

 

- Na+ (mmol/L)

142.6

141.6

141.6

141.7

 

 

 

 

- K+ (mmol/L)

4.02

4.13

4.26

4.34*

 

 

 

 

- Cl- (mmol/L)

103

103

104

103

 

 

 

 

- Ca++ (mmol/L)

2.73

2.70

2.73

2.82

 

 

 

 

- P (mmol/L)

1.89

1.76

1.64

1.75

 

 

 

 

- ASAT (U/L)

72.1

73.6

63.9

71.5

 

 

 

 

- ALAT (U/L)

38.1

42.4

51.8

49.7

53.6

56.5

53.3

 

- ALP (U/L)

145

180

231*

214

 

 

 

 

- Creat (µmol/L)

43.2

43.7

43.2

43.7

 

 

 

 

- Chol (µmol/L)

1.28

1.52

1.79*

1.79*

 

 

 

 

- Bili (µmol/L)

2.4

2.1

2.0*

9.1

 

 

 

 

- Bile ac. (µmol/L)

43.2

37.5

25.8

41.0

 

 

 

 

- Urea (mmol/L)

7.5

9.1

9.5*

9.1

 

 

 

 

Clinical Signs (% of individuals affected throughout treatment period, n=10)

- Skin

(scabs)

5-10

0

0

0

0

0

0

0

- Breathing

(rales)

0

5-10

0

0

0

0

5-10

0

- Secretion / excretion

(salivation)

0

0

100

100

0

0

100

100

- Skin

(alopecia)

0

0

0

0

0

0

0

5-10

Pathology(No. of individuals affected throughout treatment period, n=10)

No findings

7

9

0

0

8

9

3

1

Lungs

(foci)

0

0

1

0

0

0

0

0

Stomach

(foci)

0

0

2

1

0

0

0

0

Stomach

(irregular surface)

0

0

10**

10**

0

0

6*

9**

Stomach

(discoloration)

0

0

1

1

0

0

0

1

Stomach

(thickened)

0

0

0

0

0

0

0

3

Liver

(nodules)

0

0

0

1

0

0

0

0

Testes

(agenesis)

0

0

1

0

-

-

-

-

Epididymides

(nodules)

2

0

0

0

-

-

-

-

Epididymides

(agenesis)

0

0

1

0

-

-

-

-

Seminal vesicles

(size reduced)

0

1

0

0

-

-

-

-

Preputial glands

(foci)

0

0

1

0

-

-

-

-

Preputial glands

(size reduced)

0

0

0

1

-

-

-

-

Skin

(scab formation)

1

0

0

0

0

0

0

0

Skin

(alopecia)

0

0

0

0

1

0

0

1

Eyes

(right side exophthalmos)

0

0

0

1

0

0

0

0

clitoral gland

(nodules)

-

-

-

-

0

0

1

0

Clitoral gland

(foci)

-

-

-

-

1

0

1

0

Body cavities

(nodules)

0

0

0

0

0

1

0

0

* P<0.05, ** P<0.01

 

Table 5       Absolute and adjusted organ weights (F0)

 

Doses (ppm)

Control

100

300

1000

Control

100

300

1000

male (toxicity group)

female (toxicity group)

BODY WEIGHT (n=10)

Weight (g)

370

363

367

370

235

244

233

229

ADRENALS (n=5)

Weight (g)

0.065

0.064

0.067

0.070

0.079

0.073

0.082

0.075

Organ/Body Weight Ratio (%)

0.018

0.018

0.018

0.019

0.034

0.030

0.035

0.033

BRAIN (n=5)

Weight (g)

2.05

2.03

2.05

2.05

1.86

1.93

1.86

1.86

Organ/Body Weight Ratio (%)

0.56

0.57

0.55

0.55

0.79

0.79

0.80

0.81

EPIDIDYMIDES (n=10)

Weight (g)

1.153

1.155

1.169

1.146

-

-

-

-

Organ/Body Weight Ratio (%)

0.312

0.318

0.320

0.311

-

-

-

-

HEART (n=5)

Weight (g)

1.081

1.009

1.062

1.043

0.780

0.823

0.760

0.750

Organ/Body Weight Ratio (%)

0.293

0.281

0.284

0.280

0.332

0.338

0.326

0.327

KIDNEYS (n=5)

Weight (g)

2.57

2.45

2.57

2.50

1.66

1.74

1.67

1.52

Organ/Body Weight Ratio (%)

0.70

0.68

0.69

0.67

0.70

0.71

0.72

0.66

LIVER (n=5)

Weight (g)

9.79

9.31

10.43

10.75

7.47

7.86

7.59

7.73

Organ/Body Weight Ratio (%)

2.65

2.59

2.79

2.87*

3.18

3.22

3.26

3.37

OVARIES

Weight (g)

-

-

-

-

0.119

0.131

0.121

0.128

Organ/Body Weight Ratio (%)

-

-

-

-

0.051

0.054

0.052

0.056

PROSTATE (n=5)

Weight (g)

0.556

0.693

0.576

0.498

-

-

-

-

Organ/Body Weight Ratio (%)

0.150

0.193*

0.154

0.133

-

-

-

-

SEMINAL VESICLES (n=5)

Weight (g)

1.742

1.685

1.538

1.628

-

-

-

-

Organ/Body Weight Ratio (%)

0.470

0.471

0.411

0.436

-

-

-

-

SPLEEN (n=5)

Weight (g)

0.675

0.674

0.704

0.588

0.490

0.542

0.535

0.547

Organ/Body Weight Ratio (%)

0.182

0.187

0.188

0.157

0.208

0.223

0.230

0.239

TESTES (n=10)

Weight (g)

3.44

3.56

3.57

3.57

-

-

-

-

Organ/Body Weight Ratio (%)

0.93

0.98

0.98

0.97

-

-

-

-

THYMUS (n=5)

Weight (g)

0.396

0.348

0.373

0.413

0.187

0.163

0.158

0.148

Organ/Body Weight Ratio (%)

0.107

0.097

0.100

0.111

0.079

0.067

0.068

0.065

THYROID (n=5)

Weight (g)

0.029

0.030

0.029

0.030

0.026

0.021

0.027

0.020

Organ/Body Weight Ratio (%)

0.008

0.008

0.008

0.008

0.011

0.009

0.012

0.009

UTERUS

Weight (g)

-

-

-

-

0.697

0.758

0.706

0.708

Organ/Body Weight Ratio (%)

-

-

-

-

0.297

0.311

0.304

0.308

* P<0.05, ** P<0.01

  

Table 6       Summary of incidence of mating performance, fertility and gestation lengths

Group

(ppm)

# of paired males

# of females

Pre-coital interval (days)

Mating index

(%)

Pregnancy index

(%)

Gestation length (days)

Females with live offspring

Gestation index (%)

Paired

Mated

Pregnant

1-4

5-8

9-12

13-14

21

22

23

Control

10

10

10

10

10

0

0

0

100

100

10

0

0

10

100

100

10

10

10

10

10

0

0

0

100

100

10

0

0

10

100

300

10

10

10

10

10

0

0

0

100

100

10

0

0

10

100

1000

10

10

10

10

10

0

0

0

100

100

10

0

0

10

100

 

Table 7       Offspring Survival Indices

 

Group

(ppm)

Mean # of corpora lutea

Mean # of implantation sites

Mean total litter size

Number of live offspring

Viability index

(%)

Male : Female ratio

Day 1

Day 4

Control

14.2

12.9

12.3

123

121

98.4

52 : 48

100

14.2

12.6

12.1

121

121

100

48 : 52

300

15.9

13.8

13.2

132

132

100

55 : 45

1000

15.4

12.2

11.4

114

114

100

48 :52

  

Table 8       Pup weights

 

Group

(ppm)

Mean Weight Day 1

(g)

Mean Weight Day 4

(g)

Control

5.7

8.3

 

100

5.8

8.6

 

300

5.9

8.5

 

1000

6.0

8.9

 

Applicant's summary and conclusion

Conclusions:
No toxicologically significant effects on reproductive parameters were noted.

Mating, fertility and conception indices, precoital time, and number of corpora lutea and implantation sites were unaffected by treatment.

No toxicologically significant effects on developmental parameters were observed. Gestation index, duration of gestation, number of dead and Iiving pups at first Iitter check, sex ratio, postnatalloss, viability index, and early postnatal pup development (mortality, clinical signs, body weight and extern al macroscopy) were unaffected by treatment. No deficiencies in maternal care were observed. Examination of cage debris of pregnant females revealed no signs of abortion or premature birth. No signs of difficult or prolonged parturition were noted among the pregnant females.
Executive summary:

OECD 422 (2010) - In a combined repeat dose toxicity study with reproductive toxicity screening (OECD 422), n-Undecanal was administered to 30 male and 30 female Crl:WI(Han) strain rats by oral gavage at dose levels of 100, 300 and 1000 ppm for up to 7 weeks. In addition, 10 male and 20 female control rats were treated with the formulation vehicle, without test item.

A summary of adult responses to the test item are described below;

Mortality- No mortality was observed during the test.

Clinical signs- No clinical signs or dose observations related to treatment were observed during the test.

Behavioural, functional and sensory assessments- Individuals were unaffected by the treatment.

Bodyweight- Body weights and body weight gain of treated animals remained in the same range as controls over the treatment period.

Food consumption and efficiency- No toxicologically significant changes in food consumption before or after allowance for body weight were noted.

Haematology- Haematological parameters of treated rats were considered not to have been affected by treatment.

Blood chemistry- A statistically significant higher potassium value was measured for males at 1000 mg/kg/day, but the mean remained within the range considered normal for rats of this age and strain. In females at 1000 mg/kg/day, a higher mean cholesterol level was recorded (being slightly outside the range considered normal for rats of this age and strain) along with notably higher bile acid levels in some females (means not statistically significant). Given that these changes were generally slight in nature, were not present as a group response (bile acids) and occurred in the absence of supportive morphological changes, these were considered to be of no toxicological relevance.

Other statistically significant variations in clinical biochemistry parameters were considered to be of no toxicological relevance as these occurred in the absence of a treatment-related distribution and/or remained within the range considered normal for rats of this age and strain. These variations consisted of higher or lower alkaline phosphatase activity (ALP) in males and females at 300 mg/kg/day, lower total bilirubin level in males at 300 mg/kg/day, higher urea level in males at 300 mg/kg/day, higher cholesterol level in males at 300 and 1000 mg/kg/day, and lower urea level in females at 1000 mg/kg/day.

Necropsy- The following necropsy findings were considered to be related to treatment with the test substance:

- An irregular surface of the forestomach in all males at 300 and 1000 mg/kg/day, and in 6 females at 300 mg/kg/day and nine females at 1000 mg/kg/day.

- Foci (black or red) in the forestomach in two males at 300 mg/kg/day.

- Yellowish discolouration of the forestomach in one male at 300 mg/kg/day and one female at 1000 mg/kg/day.

- Reddish discolouration reddish foci in the glandular mucosa in two males at 1000mg/kg/day.

- Thickened glandular mucosa in three females at 1000 mg/kg/day.

Incidental findings among control and treated animals included reddish foci on the lungs, a nodule on the liver, epididymides or clitoral glands, agenesis of the testes and epididymides, reduced size of the seminal vesicles or preputial glands, tan foci on the preputial glands, a greenish/red-brown foci on the clitoral glands, a yellowish nodule on the uterine adipose tissue, scab formation on the skin, alopecia and exophthalmus. The incidence of these findings was within the background range of findings that are encountered among rats of this age and strain, and the incidence did not show a dose-related trend. These necropsy findings were therefore considered to be of no toxicological significance.

Organ weights- A higher liver weight and liver to body weight ratio, and lower prostate and prostate to body weight ratio was noted for males at 1000 mg/kg/day (statistically significant for liver to body weight ratio only). These changes were only slightly outside the normal range for rats of this age and strain.

The higher prostate to body weight ratio of males at 100 mg/kg/day occurred in the absence of a dose-related trend, and the mean remained within the range considered normal for rats of this age and strain. No toxicological relevance was ascribed to this change. Other organ weights and organ to body weight ratios among the dose groups were similar to control levels.

The higher liver weight and liver to body weight ratio, and lower prostate and prostate to body weight ratio for males at 1000 mg/kg/day was not supported by any histopathological changes. Moreover, since these changes were slight in nature (only slightly outside the normal range), these were considered to be of no toxicological relevance.

Histopathological changes-Histopathological changes were confined to the stomach, and consisted of Iymphogranulocytic inflammation and hyperplasia of the squamous epithelium of the forestomach in both sexes at 100, 300 and 1000 mg/kg/day, and ulcer formation in the forestomach in three males at 300 mg/kg/day (correlating to red/black foci in the forestomach of two males) and in one male at 1000 mg/kg/day. Hyperplasia ofthe squamous epithelium was the microscopic correlate to the irregular surface and yellowish discolouration of the forestomach recorded at necropsy at 300 and 1000 mg/kg/day. Congestion of the glandular stomach (correlating to red discolouration of of the glandular mucosa) was observed in two males at 1000 mg/kg/day. There was no microscopic correlate to thickening of the glandular mucosa recorded in three females at 1000 mg/kg/day.

A summary of reproductive and developmental findings is described below;

Reproduction data- No toxicologically significant effects on reproductive parameters were noted. Mating, fertility and conception indices, precoital time, and number of corpora lutea and implantation sites were unaffected by treatment.

Developmental data- No toxicologically significant effects on developmental parameters were observed. Gestation index, duration of gestation, number of dead and living pups at first litter check, sex ratio, postnatal loss, viability index, and early postnatal pup development (mortality, clinical signs, body weight and extern al macroscopy) were unaffected by treatment. No deficiencies in maternal care were observed. Examination of cage debris of pregnant females revealed no signs of abortion or premature birth. No signs of difficult or prolonged parturition were noted among the pregnant females.  Incidental clinical symptoms of pups consisted of small size, red spots on the head/nose and a pale appearance. No relationship with treatment was established for these observations and they were considered to be of no toxicological relevance. A total of three pups of the control group, and one pup each at 100, 300 and 1000 mg/kg/day were found dead or missing during lactation. No toxicological relevance was ascribed to these dead/missing pups since the mortality incidence did not show a dose-related trend and remained within the range considered normal for pups of this age. Incidental macroscopic findings among pups found dead included autolysis, cannibalism and absence of milk in the stomach. Some surviving pups were small in size. The nature and incidence of these findings remained within the range considered normal for pups of this age, and were therefore considered to be unrelated to treatment.

Based on these considerations, a No Observed Adverse Effect Level (NOAEL) for reproductive and developmental toxicity was therefore concluded to be 1000 mg/kg/day.

This combined toxicity study with reproduction screening in the rat is acceptable and satisfies the guideline requirements for an OECD 422 in the rat

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