Registration Dossier
Registration Dossier
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EC number: 403-830-5 | CAS number: 89331-94-2 B 290; BK 400; CK 34; DIBUTYL-N-102; DX-20; FAT NR. 40391/A; FLUORAN BLACK BD 869; FLUORAN SCHWARZ BD 869; NOIR FLUORANE BD 869; ODB-2; PSD-290; SENOR-2; TG-31; TH-108; WINCON-2
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data

Toxicity to reproduction
Administrative data
- Endpoint:
- one-generation reproductive toxicity
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 1992-10-05 to 1993-03-03
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study with acceptable restrictions
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 1 993
- Report date:
- 1993
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 415 [One-Generation Reproduction Toxicity Study (before 9 October 2017)]
- Version / remarks:
- 1983-05
- Deviations:
- no
- GLP compliance:
- yes
- Limit test:
- no
Test material
- Reference substance name:
- 6'-(dibutylamino)-3'-methyl-2'-(phenylamino)spiro[isobenzofuran-1(3H),9-(9H)-xanthen]-3-one
- EC Number:
- 403-830-5
- EC Name:
- 6'-(dibutylamino)-3'-methyl-2'-(phenylamino)spiro[isobenzofuran-1(3H),9-(9H)-xanthen]-3-one
- Cas Number:
- 89331-94-2
- Molecular formula:
- C35H36N2O3
- IUPAC Name:
- 6'-(dibutylamino)-3'-methyl-2'-(phenylamino)-3H-spiro[2-benzofuran-1,9'-xanthen]-3-one
Constituent 1
Test animals
- Species:
- rat
- Strain:
- Sprague-Dawley
- Remarks:
- CD (SD) BR VAF/Plus strain
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Age at study initiation: (P) male: 4 wks, female: 7 - 8 wks
- Weight at study initiation: (P) Males: 72 - 95 g; Females:174 - 196 g
- Housing: During the pre-mating period, males and females were housed separately, four to a cage. During the mating period, animals were housed on the basis of one male to one female in plastic breeding cages (North Kent Plastics, RM-2 type). At the end of the mating period the males were rehoused with their former cage mates in metal cages (Biotech®) and the females remained in individual breeding cages (North Kent Plastics, RM-2 type) for the birth and rearing of young. Following the sacrifice of weanlings, females were rehoused with their former cage mates in metal cages (Biotech®).
- Diet: Biosure Laboratory Animal Diet No. 1, ad libitum
- Water: tap water, ad libitum
- Acclimation period: 12 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 ± 4
- Humidity (%): 45 ± 17
- Photoperiod (hrs dark / hrs light): 12/12
Administration / exposure
- Route of administration:
- oral: gavage
- Vehicle:
- other: 1 % methylcellulose
- Details on exposure:
- PREPARATION OF DOSING SOLUTIONS: 1 mL/100 gram bodyweight
The test substance was ground in a mortar with a small amount of the vehicle (1 % methylcellulose) until a smooth paste was formed. The formulation was then gradually made up to volume and mixed using a high shear homogeniser to give the concentration used for the highest dosage (10.0% w/v). A sequence of suspensions was then made from the highest concentration to give concentrations of 2.5 and 0.625% w/v for the lower dosages. A constant dose volume of 1 mL/100 g bodyweight was assumed throughout.
DIET PREPARATION
- Rate of preparation of diet: daily
- Details on mating procedure:
- - M/F ratio per cage: 1/1
- Length of cohabitation: 20 days
- Proof of pregnancy: Sperm in vaginal smear. - Analytical verification of doses or concentrations:
- yes
- Duration of treatment / exposure:
- Treatment commenced when the males were approximately 6 weeks of age (designated time 0 with the succeeding week designated Week 1 of the study) and continued for 10 weeks prior to mating, through the mating, gestation and lactation periods and up to sacrifice after weaning of pups. The dose volume was calculated for individual animals on the first day of treatment and adjusted at weekly intervals throughout the study.
Sexually mature females were treated for 2 weeks prior to mating (treatment commenced after the males had been treated for 8 weeks). Dosing continued throughout mating, gestation and lactation and up to sacrifice after weaning. The dose volume was calculated for individual animals on the first day of treatment and adjusted at weekly intervals up to and including the second week after pairing. Dose volumes remained constant for each animal thereafter until at least 7 days after Day 21 post partum when volumes were readjusted at weekly intervals up to sacrifice. - Frequency of treatment:
- Daily
Doses / concentrationsopen allclose all
- Dose / conc.:
- 0 mg/kg bw/day
- Dose / conc.:
- 62.5 mg/kg bw/day
- Dose / conc.:
- 250 mg/kg bw/day
- Dose / conc.:
- 1 000 mg/kg bw/day
- No. of animals per sex per dose:
- 24
- Control animals:
- yes, concurrent vehicle
Examinations
- Parental animals: Observations and examinations:
- CAGE SIDE OBSERVATIONS: Yes
- Time schedule: All animals were regularly handled and examined for obvious changes or signs of reaction to treatment.
BODY WEIGHT: Yes
- Time schedule: During the mating period, all females were weighed daily throughout and daily weighing continued until parturition. Weights are reported for Days 0, 7, 14, 17 and 20 of pregnancy. Weights of pregnant animals without a positive indication of mating are reported for appropriate days taken retrospectively from birth. Dams that littered were weighed on Days 0, 7, 14 and 21 post partum.
FOOD CONSUMPTION:
- Food consumption: Yes
- Time schedule for examinations: Food intake of rats was recorded weekly for the week inunediately prior to treatment and throughout the pre-mating phase. During the pre-mating phase, food conversion ratios were calculated.
WATER CONSUMPTION: Yes
- Time schedule for examinations: Water consumption was measured on a daily basis during the pre-dose week for both sexes, during the first two weeks of treatment for males and for the two weeks prior to mating (males and females). Water consumption was also recorded from allocation until initiation of treatment (Week -1 for males, Week 8 for females).
OTHER:
- Prior to treatment and scheduled terminal sacrifice, all surviving animals were subjected to an ophthalmoscopic examination. - Litter observations:
- STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: yes
PARAMETERS EXAMINED
- The following parameters were examined in F1 offspring:
Number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, weight gain, physical or behavioural abnormalities
ASSESSMENT OF DEVELOPMENTAL NEUROTOXICITY:
- Offspring surface righting reflex, offspring startle reflex, offspring air righting reflex, offspring pupil reflex - Postmortem examinations (parental animals):
- SACRIFICE
- Male animals: All surviving animals
- Maternal animals: All surviving animals
GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations including the cervical, thoracic, and abdominal viscera.
HISTOPATHOLOGY / ORGAN WEIGHTS
The tissues indicated were prepared for microscopic examination and weighed: prostate, seminal vesicles, coagulating gland, epididymides, testes, pituitary, uterus, cervix, vagina, ovaries, oesophagus
The following tissues were additionally weighed: brain, thymus, heart, lungs, liver, kidneys, adrenals - Postmortem examinations (offspring):
- SACRIFICE
- The F1 offspring were sacrificed at 21 days of age.
GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations including the cervical, thoracic, and abdominal viscera. - Statistics:
- All statistical analyses were carried out separately for males and females.
Significance tests, employing analysis of variance following by an intergroup comparison with control, were performed on the following parameters and results are presented in relevant tables of the report: weekly bodyweight and female bodyweight change during pregnancy and lactation, food and water consumption, litter data and organ weights. Dependent on the heterogeneity of variance between treatment groups, parametric tests (analysis of variance (Snedecor and Cochran, 1967) followed by William’s’ test (Williams, 1971/2)) or non-parametric tests (Kryskal-Wallis (Hollander and Wolfe, 1968) followed by Shirley’s test (Shirley, 1977)) were used to analyze these data, as appropriate. For bodyweight and bodyweight change, the analyses were carried out using the individual animal as the experimental unit. Data relating to food and water consumption were analyses on a cage basis. For litter data, the basic sample unit was the litter and, due to the preponderance of non-normal distributions, nonparametric analyses were routinely used. Organ weight data were analyzed using body weight at post mortem as covariate, to allow for differences in bodyweight which may have influenced organ weight values. Where 75% or more of the values for a given variable were the same, a Fisher’s exact test (Fisher, 1950) was used, when considered necessary. All significant (i.e. P < 0.05) intergroup differences from the control are reported and were supported by a significant analysis of variance (P < 0.05). - Reproductive indices:
- Pregnancy rate was determined as the percentage of surviving paired females that became pregnant.
Results and discussion
Results: P0 (first parental generation)
General toxicity (P0)
- Clinical signs:
- no effects observed
- Description (incidence and severity):
- There were no treatment-related clinical signs observed during the study.
- Mortality:
- mortality observed, non-treatment-related
- Description (incidence):
- One female in the high dose group died in week 11 without previous clinical signs. The pathological examination revealed moist red staining of the perioral and perinasal fur and severely congested, uncollapsed lungs, but no signs of an intubation error. One male of the high dose group was sacrificed in week 13 after having been observed continually leaning to one side for nine days. Prior to sacrifice the animal appeared uncoordinated and continually rolled over. No abnormalities were detected during macroscopic post mortem examination.
One female in the low dose group was found dead in week 14, with red salivation and vaginal discharge. No previous clinical signs were recorded. Post mortem findings included enlarged cervical lymph nodes, congested thymus and lungs, gaseous distension, and blood in the stomach, as well as dark contents in the small intestine. The uterus contained recently dead foetuses and placentae. Thirteen pups had been born, all were cold and unfed when the dam was found.
As there were no trends or consistent pathology findings among these animals, the mortalities were considered to be coincidental and not treatment related.
Three female rats, one from control, two from the high dose group, were either found dead or were sacrificed an the second day of treatment following loss of body tone and laboured respiration. Post mortem examination revealed all three to have perforated oesophagus. The clinical signs and necropsy findings were consistent with accidental intubation errors occurring during the dosing procedure. As one of the animals receiving the high dose was found dead within 24 h of the first dose, a replacement animal was selected for the duration of the study from this day. - Body weight and weight changes:
- no effects observed
- Description (incidence and severity):
- Weekly bodyweight gain for males and femals was generally similar for all groups (P > 0.05).
Bodyweight change for femals during pregnancy and lactation was uneffected by treatment (P > 0.05). - Food consumption and compound intake (if feeding study):
- no effects observed
- Description (incidence and severity):
- Food consumption for both sexes recorded during the pre-mating period (to week 10) was essentially similar in all groups (cumulative intake P > 0.05).
- Food efficiency:
- no effects observed
- Description (incidence and severity):
- The efficieny of food utilisation during the pre-mating period, although slightly variable, was unaffected by treatment.
- Water consumption and compound intake (if drinking water study):
- no effects observed
- Description (incidence and severity):
- Intergroup variation in water consumption during the periods recorded for males and females showed no dose relationship and cumulative values did not attain statistical significance (P > 0.05).
- Ophthalmological findings:
- no effects observed
- Description (incidence and severity):
- The type of ocular lesions seen were consistent with the age and strain of rat. There were no treatment-related changes.
- Haematological findings:
- not examined
- Clinical biochemistry findings:
- not examined
- Urinalysis findings:
- not examined
- Behaviour (functional findings):
- not examined
- Immunological findings:
- not examined
- Organ weight findings including organ / body weight ratios:
- effects observed, non-treatment-related
- Histopathological findings: non-neoplastic:
- no effects observed
- Description (incidence and severity):
- No microscopic findings were detected in the tissues examined.
- Histopathological findings: neoplastic:
- not examined
- Other effects:
- not examined
Reproductive function / performance (P0)
- Reproductive function: oestrous cycle:
- not examined
- Reproductive function: sperm measures:
- not examined
- Reproductive performance:
- no effects observed
- Description (incidence and severity):
- Mating performance, as assessed by the incidence and distribution of successful matings, the median pre-coital time and the type of smear recorded on the day of conception, was not adversely affected by treatment. The pregnancy rate was 100 % for all groups with most females conceiving within the first four days after the start of the pairing, corresponding with the expected length of an oestrus cycle. The duration of the pregnancy was similar for all groups (P > 0.05).
Effect levels (P0)
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- >= 1 000 mg/kg bw/day
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: no adverse effects were observed up to the highest dose
Target system / organ toxicity (P0)
- Key result
- Critical effects observed:
- no
Results: F1 generation
General toxicity (F1)
- Mortality / viability:
- no mortality observed
- Organ weight findings including organ / body weight ratios:
- no effects observed
- Description (incidence and severity):
- Intergroup differences in organ weights, adjusted for final bodyweight as appropriate, were only slight (P>0.05) and revealed no clear or consistent changes in either sex which could be attributed to treatment.
- Gross pathological findings:
- no effects observed
- Description (incidence and severity):
- The macroscopic examination performed at terminal autopsy revealed no treatment-related changes.
- Histopathological findings:
- no effects observed
- Other effects:
- no effects observed
- Description (incidence and severity):
- Mean ages of attainment of surface and air righting reflexes, startle response and presence of pupil reflex at day 20 post partum were similar for treated and control groups.
Details on results (F1)
Among dams rearing young to weaning, mean values for implantation rates, pup survival, pup growth and associated sex ratio at birth and Day 21 post partum were similar for all groups (P>0.05).
Effect levels (F1)
- Key result
- Dose descriptor:
- NOAEL
- Generation:
- F1
- Effect level:
- >= 1 000 mg/kg bw/day
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: no adverse effects were observed up to the highest dose
Target system / organ toxicity (F1)
- Key result
- Critical effects observed:
- no
Overall reproductive toxicity
- Key result
- Reproductive effects observed:
- no
- Lowest effective dose / conc.:
- 1 000 mg/kg bw/day
- Treatment related:
- no
Any other information on results incl. tables
Table 1: Summary of Adult Performance
Category |
No of Animals in Group/Dosage [mg/kg bw/day] |
|||
Control |
62.5 |
250 |
1000 |
|
Males |
||||
Initial Group Size |
24 |
24 |
24 |
24 |
Mortality at Day 19 of Pairing |
- |
- |
- |
1** |
Inducing Pregnancy |
23* |
24 |
24 |
23 *,** |
Females |
||||
Initial Group Size |
24 |
24 |
24 |
24 |
Mortaliy: |
|
|
|
|
Pre-Pairing |
1 |
- |
- |
1 |
Day 3 of Pairing |
- |
- |
- |
1 |
Day 1 post partum |
- |
1 |
1 |
- |
Rearing young to weaning |
23 |
22 |
23 |
22 |
* One male not paired as female partner previously killed ** Male had induced pregnancy prior to sacrifice |
Table 2: Group mean Bodyweight Values [g]
Week |
Group/Dosage [mg/kg bw/day] |
|||||||
Male |
Female |
|||||||
Control |
62.5 |
250 |
1000 |
Control |
62.5 |
250 |
1000 |
|
-1 |
137 |
138 |
139 |
138 |
|
|
|
|
0 – T male |
203 |
202 |
204 |
203 |
|
|
|
|
1 |
264 |
263 |
266 |
261 |
|
|
|
|
2 |
322 |
321 |
324 |
318 |
|
|
|
|
3 |
369 |
371 |
373 |
363 |
|
|
|
|
4 |
406 |
410 |
412 |
400 |
|
|
|
|
5 |
439 |
445 |
449 |
434 |
|
|
|
|
6 |
467 |
472 |
480 |
462 |
|
|
|
|
7 |
492 |
498 |
506 |
486 |
221 |
220 |
220 |
222 |
8 – T female |
514 |
520 |
531 |
510 |
244 |
246 |
242 |
243 |
9 |
523 |
532 |
542 |
521 |
255 |
259 |
255 |
256 |
10 P |
541 |
550 |
561 |
537 |
274 |
276 |
272 |
277 |
11 |
548 |
557 |
570 |
550 |
317 |
324 |
315 |
317 |
12 |
566 |
573 |
587 |
564 |
359 |
364 |
358 |
361 |
13 |
574 |
576 |
594 |
566 |
435 |
436 |
432 |
440 |
14 |
588 |
591 |
603 |
577 |
356 |
359 |
357 |
367 |
15 |
606 |
608 |
620 |
590 |
381 |
381 |
381 |
375 |
16 |
621 |
625 |
641 |
604 |
367 |
365 |
360 |
368 |
17 |
634 |
639 |
654 |
617 |
332 |
336 |
327 |
336 |
18 |
|
|
|
|
337 |
343 |
332 |
341 |
19 |
|
|
|
|
347 |
354 |
343 |
354 |
20 |
|
|
|
|
351 |
358 |
350 |
358 |
21 |
|
|
|
|
355 |
361 |
353 |
361 |
Bodyweight gain [g/rat] |
||||||||
Weeks |
431 |
437 |
451 |
415 |
111 |
115 |
111 |
118 |
S.D. |
78.5 |
52.4 |
77.9 |
63.4 |
15.1 |
19.8 |
14.9 |
19.2 |
% Control |
- |
101 |
105 |
96 |
- |
104 |
100 |
106 |
S.D. Standard Deviation T Start of treatment P Animals paired |
Table 3: Group mean Food Consumption Values [g/rat/week]
Week |
Group/Dosage [mg/kg bw/day] |
|||||||
Male |
Female |
|||||||
Control |
62.5 |
250 |
1000 |
Control |
62.5 |
250 |
1000 |
|
-1 |
154 |
152 |
155 |
151 |
|
|
|
|
1 – T male |
178 |
176 |
173 |
170 |
|
|
|
|
2 |
199 |
197 |
196 |
190 |
|
|
|
|
3 |
205 |
205 |
204 |
196 |
|
|
|
|
4 |
204 |
207 |
209 |
200 |
|
|
|
|
5 |
208 |
210 |
212 |
201 |
|
|
|
|
6 |
212 |
212 |
215 |
205 |
|
|
|
|
7 |
207 |
212 |
212 |
206 |
|
|
|
|
8 |
204 |
206 |
206 |
202 |
146 |
148 |
148 |
145 |
9 – T female |
206 |
209 |
206 |
203 |
147 |
148 |
150 |
144 |
10 |
205 |
209 |
207 |
201 |
152 |
150 |
150 |
149 |
Cumulative intake [g/rat] |
||||||||
Male: 1 – 10 Female: 9 – 10 |
2026 |
2043 |
2042 |
1975 |
299 |
298 |
301 |
294 |
S.D. |
49.5 |
67.3 |
103.3 |
58.6 |
14.1 |
13.6 |
7.6 |
8.0 |
% Control |
- |
101 |
101 |
97 |
- |
100 |
101 |
98 |
S.D. Standard Deviation T first week of treatment
|
Table 4: Group mean Food Conversion Ratio Values (= food consumption/bw gain)
Week |
Group/Dosage [mg/kg bw/day] |
|||||||
Male |
Female |
|||||||
Control |
62.5 |
250 |
1000 |
Control |
62.5 |
250 |
1000 |
|
1 – T male |
2.9 |
2.9 |
2.8 |
2.9 |
|
|
|
|
2 |
3.4 |
3.4 |
3.4 |
3.3 |
|
|
|
|
3 |
4.3 |
4.1 |
4.2 |
4.3 |
|
|
|
|
4 |
5.6 |
5.2 |
5.4 |
5.4 |
|
|
|
|
5 |
6.2 |
6.1 |
5.7 |
6.0 |
|
|
|
|
6 |
7.8 |
7.7 |
7.1 |
7.1 |
|
|
|
|
7 |
8.2 |
8.0 |
7.9 |
8.6 |
|
|
|
|
8 |
9.3 |
9.3 |
8.3 |
8.5 |
|
|
|
|
9 – T female |
22.1 |
18.3 |
18.4 |
18.7 |
13.0 |
11.6 |
11.3 |
10.7 |
10 |
11.4 |
11.7 |
11.1 |
12.7 |
7.7 |
8.9 |
8.7 |
7.1 |
Mean values |
||||||||
Male: 1 – 10 Female: 9 – 10 |
6.0 |
5.9 |
5.7 |
5.9 |
9.7 |
10.0 |
9.9 |
8.5 |
T first week of treatment |
Table 5: Group mean Water Consumption Values [g/rat/week]
Week |
Group/Dosage [mg/kg bw/day] |
|||||||
Male |
Female |
|||||||
Control |
62.5 |
250 |
1000 |
Control |
62.5 |
250 |
1000 |
|
-1 |
184 |
177 |
189 |
183 |
|
|
|
|
1 – T male |
206 |
199 |
218 |
209 |
|
|
|
|
2 |
232 |
229 |
252 |
229 |
|
|
|
|
8 |
|
|
|
|
200 |
218 |
215 |
205 |
9 T – female |
257 |
265 |
291 |
264 |
190 |
209 |
207 |
199 |
10 |
255 |
261 |
278 |
269 |
200 |
222 |
210 |
212 |
Cumulative intake [g/rat] |
||||||||
Week 1 - 2 |
438 |
428 |
470 |
438 |
|
|
|
|
S.D. |
30.4 |
21.9 |
28.7 |
27.1 |
|
|
|
|
% Control |
- |
98 |
107 |
100 |
|
|
|
|
Week 9 - 10 |
512 |
526 |
568 |
533 |
391 |
431 |
416 |
413 |
S.D. |
59.6 |
71.0 |
71.1 |
48.6 |
33.1 |
28.0 |
32.2 |
29.5 |
% Control |
- |
103 |
111 |
104 |
- |
110 |
106 |
106 |
S.D. Standard Deviation T first week of treatment
|
Table 6: Summary of Ophthalmoscopic Observations
Category |
No of Animals in Group/Dosage [mg/kg bw/day] |
|||
Control |
62.5 |
250 |
1000 |
|
Males |
||||
Pre-treatment (Week -1) |
||||
Examined |
24 |
24 |
24 |
24 |
Vitreous: hyaloid remnants |
4 |
1 |
3 |
5 |
Pre-sacrifice (Week 17) |
||||
Examined |
24 |
24 |
24 |
23 |
Lens: posterior capsular opacity |
- |
- |
1 |
- |
Vitreous: hyaloid remnants |
1 |
- |
- |
- |
Retina: apparent hyperreflectivity |
1 |
- |
- |
1 |
Females |
||||
Pre-treatment (Week 8) |
||||
Examined |
24 |
24 |
24 |
24 |
Vitreous: hyaloid remnants |
6 |
9 |
5 |
5 |
haemorrhage |
- |
1 |
- |
- |
Pre-sacrifice (Week 21) |
||||
Examined |
23 |
23 |
24 |
22 |
Vitreous: hyaloid remnants |
- |
1 |
2 |
1 |
Retina: abnormality of optic nerve head |
1 |
- |
- |
1 |
Table 7: Group Values of Mating Performance
|
Group/Dosage [mg/kg bw/day] |
|||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Control |
62.5 |
250 |
1000 |
|||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
No of mating pairs |
23 |
24 |
24 |
22 d |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
No conceiving on Day |
|
|||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
1 |
12 |
9 |
5 |
7 |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
2 |
3 |
2 |
10 |
7 |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
3 |
2 |
8 r |
8 |
3 |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
4 |
6 |
4 |
- |
2 |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
6 |
- |
- |
- |
2 |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
7 |
- |
- |
- |
1 |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
15 |
- |
1 |
- |
- |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
U |
- |
- |
1 r |
- |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Total |
23 |
24 |
24 |
22 |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Pregnancy rate [%] |
100 |
100 |
100 |
100 |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Median pre-coital time [days] |
1.0 |
3.0 |
2.0 |
2.0 |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Cell type at conception(from vaginal smear) |
|
|||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
S |
22 |
23 r |
23 |
18 |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
C |
- |
1 |
- |
2 |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
L (Day 1) |
1 |
- |
- |
- |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
L |
- |
- |
- |
2 |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
U |
- |
- |
1 r |
- |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Duration of pregnancy [days] |
|
|||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
21 |
3 |
4 |
4 |
2 |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
22 |
14 |
16 |
17 |
16 |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
23 |
5 |
3 |
2 |
2 |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
U |
1 |
- |
- |
2 |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Mean |
22.1 |
22.0 |
21.9 |
22.0 |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
d Excludes one dam found dead on day 3 of pairing r Includes one dam with total resorption U Undetermined Predominant cell type: S Sperm C Cornified epithelial L Leucocyte Table 8: Group mean Values of Litter Data
Table 9: Group mean Values of the Sex Ratio
Table 10: Group mean Values of pre-weaning Development
Table 11: Absolute Organ Weights
|
Applicant's summary and conclusion
- Conclusions:
- Based on the result obtained, this study indicated that dosages of 62.5, 250 or 1000 mg/kg/d were without adverse effect on the growth and reproductive capacity of male and female rats or the development of their offspring. The dosage of the test substance at which no signs of toxicity were recorded is therefore considered to be 1000 mg/kg/d.
- Executive summary:
In an 1-generation reproduction study according to OECD guideline 415 (Huntingdon, 1993), the test substance was administered to 24 Sprague Dawley rats/sex/dose by gavage at dose levels of 62.5, 250 or 1000 mg/kg/d.
Each group consisted of 24 male and 24 female rats. Animals from all groups were observed for clinical signs, mortality and changes in body weight, food consumption, water consumption and ophthalmoscopy during the experiment. Furthermore pregnancy rate, mating performance (7 days prior to mating and daily during 20 day mating period), duration of pregnancy and different litter data (offspring surface righting reflex, offspring startle reflex, offspring air righting reflex, offspring pupil reflex) were examined.
After a minimum of 10 weeks of treatment, females were cohabitated with males in a 1:1 (one male to one female) ratio. The number, weight, survivability and mortality of pups were observed during the lactation period. Physical signs of postnatal development were observed daily until the criterion was met.
The animals were subjected to detailed necropsy at sacrifice and specified organs were weighed. Histopathological examination of parents was initially carried out for the preserved organs including gross lesions. For F1 weanlings, macroscopical examinations were also performed.
There were no treatment related clinical signs observed during the study. One female receiving 1000 mg/kg/day was found dead during week 11. Post mortem revealed moist red staining of the perioral and perinasal fur and severely congested, uncollapsed lungs but no apparent signs of intubation error. One male receiving 1000 mg/kg/day was sacrificed week 13. Prior to sacrifice this animal appeared uncoordinated and continually rolled over, however there were no macroscopic abnormalities. One female in the low dose group was found dead in week 14. Post mortem findings included enlarged cervical lymph nodes, congested thymus and lungs, gaseous distension of and blood in the stomach, dark contents in the small intestines. Uterus contained recently dead foetuses and placentae; 13 pups had been born. These mortalities were considered to be coincidental and not related to treatment. A further three animals died from accidental intubation errors. No other effects were observed in parental animals with regard to body weight, food consumption, food conversion ratio, water consumption and ophthalmoscopy.
Mating performance, as assessed by the incidence and distribution of successful matings, the median pre-coital time and the type of smear recorded on the day of conception, was not adversely affected by treatment. The pregnancy rate was 100 % for all groups with most females conceiving within the first four days after the start of the pairing, corresponding with the expected length of an oestrus cycle. The duration of the pregnancy was similar for all groups (P > 0.05).
Two females resorbed their single implant. This incidence was considered not to be treatment related. Among dams rearing young to weaning, mean values for implantation rates, pup survival, pup growth and associated sex ratio at birth and Day 21 post partum were similar for all groups.
The macroscopic examination performed at terminal autopsy of surviving adults and offspring revealed no treatment-related changes. No microscopic findings were detected in the tissues examined.
Based on the result obtained, this study indicated that dosages of 62.5, 250, or 1000 mg/kg/day were without adverse effect on the growth and reproductive capacity of male and female rats or the development of their offspring. The dosage of the test substance at which no signs of toxicity were recorded is therefore considered to be 1000 mg/kg/day.
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