Registration Dossier

Toxicological information

Toxicity to reproduction

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Administrative data

Endpoint:
two-generation reproductive toxicity
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2002
Report Date:
2002

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 416 (Two-Generation Reproduction Toxicity Study)
Version / remarks:
1983
Deviations:
no
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 416 (Two-Generation Reproduction Toxicity Study)
Version / remarks:
2001
Deviations:
yes
Remarks:
the conduct of the study was consistent in all important aspects to the OECD 416 (2001) except that the examination of the oestrus cycle and sperm parameters are missing.
GLP compliance:
yes
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Test material form:
solid - liquid: aqueous solution
Details on test material:
- Name of test material: DOPA-Glycinate

Test animals

Species:
rat
Strain:
other: Wistar rats Crl: (WI) BR (outbred, SPF-Quality)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River, Sulzfeld, Germany
- Age at study initiation: P-females: 15 weeks, P-males: 7 weeks
- Weight at study initiation: P-females: 219–299g, P-males: 236–292g
- Housing: premating - in groups of max. 4; during mating - 1:1; after mating - individually; offsprong were kept with the dam until weaning
- Diet (e.g. ad libitum): standard pelleted diet, ad libitum
- Water (e.g. ad libitum): tap water, ad libitum
- Acclimation period: 13-20 d

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21±3
- Humidity (%): 30-70
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Details on mating procedure:
- M/F ratio per cage: 1:1
- Length of cohabitation: max. 3 weeks
- Proof of pregnancy: vaginal plug or sperm in vaginal smear referred to as day 0 of pregnancy
- Further matings after three unsuccessful weeks: no
- After successful mating each pregnant female was caged individually
Analytical verification of doses or concentrations:
yes
Duration of treatment / exposure:
P-males: 10 weeks prior to mating up to termination.
P-females: 2 weeks prior to mating, during mating, pregnancy and lactation.
F1-males: from weaning onwards for at least 10 weeks prior to mating up to termination.
F1 females: from weaning onwards for at least 10 weeks prior to mating, during mating, pregnancy and lactation.
F2-pups were not treated.
Frequency of treatment:
daily
Doses / concentrations
Remarks:
Doses / Concentrations:
0, 10, 30 and 100 mg/kg bw based on test substance as manufactured (20% aqueous solution)
Basis:
actual ingested
No. of animals per sex per dose:
P-generation: 24 animals of each sex per group
F1-generation: 22 to 24 animals of each sex per group
Control animals:
yes, concurrent vehicle

Examinations

Parental animals: Observations and examinations:
Clinical signs: Yes, daily

Body weight: Yes; weekly
Mated females were weighed on gestation days 0, 7, 14 and 21 and during lactation on the same days as the weighing of the litters (days 1, 4, 7, 14, 21 and 25).

Food consumption: Yes, weekly
Food consumption of mated females was measured on gestation days 0, 7, 14 and 21 and during lactation on the same days as the weighing of the litters (days 1, 4, 7, 14, 21 and 25).

Water consumption: Subjective appraisal was maintained during the study, but no quantitative investigation performed since no effect on water consumption was suspected.

Oestrous cyclicity (parental animals):
Not determined
Sperm parameters (parental animals):
Not determined
Litter observations:
STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: yes
- If yes, maximum of 8 pups/litter (4/sex/litter as nearly as possible); excess pups were killed and discarded.

PARAMETERS EXAMINED
The following parameters were examined in [F1 / F2 ] offspring:
number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, weight gain

GROSS EXAMINATION OF DEAD PUPS:
yes, for external and internal abnormalities; possible cause of death wasnot determined for pups born or found dead.
Postmortem examinations (parental animals):
SACRIFICE
F0-males were killed as soon as possible after confirmation of pregnancy of F0-females. F0-females were killed as soon as possible after the lactation period.
On day 4 of lactation or shortly thereafter, F1-offspring and F2-offspring eliminated for standardisation of the litter were killed. F1-offspring not selected for mating were killed as soon as possible after weaning. F1-offspring selected for mating were killed as soon as possible after weaning of the F2-offspring.

MACROSCOPIC EXAMINATION
All parental animals (P and F1) were subjected to macroscopic examination of the cranial, thoracic, abdominal tissues and organs, with special attention to the reproductive organs and abnormalities.
Samples of the following tissues and organs were fixed in 4% formaldehyde: all gross lesions, cervix, coagulation gland, epididymides, ovaries, pituitary gland, prostate, seminal vesicles, stomach, testes, uterus, vagina.
Postmortem examinations (offspring):
F1-offspring not selected for mating and F2-offspring:
Offspring found dead or killed before day 14 of lactation were sexed and externally examined if practically possible. If possible, defects or cause of death were evaluated. The stomach was examined for the presence of milk. No pups were preserved for further examination.
Offspring found dead or killed on or after day 14 of lactation were subjected to macroscopic examination of the cranial, thoracic and abdominal tissues and organs. Descriptions of all macroscopic abnormalities were recorded. If possible, defects or cause of death were evaluated. No pups were preserved for further examination.
Statistics:
Body weight, body weight gain, food consumption, relative food consumption, pup weight: Dunnett-Test (variables could be assumed to follow a normal distribution)
Implantation sites, living pups at 1st litter check: Steel-test (data cannot be assumed follow a normal distribution)
Macroscopic findings, fertility index, conception rate, gestation index, percentage mating, breeding data: Fisher’s Exact Test
Reproductive indices:
Percentage mating
Fertility index
Conception rate
Gestation index
Percentage live males at first litter check
Percentage live females at first litter check
Percentage of postnatal loss days 0-4 post partum
Percentage of breeding loss day 5 until weaning
Percentage live males at weaning
Percentage live females at weaning
Viability index
Weaning index

Results and discussion

Results: P0 (first parental animals)

General toxicity (P0)

Clinical signs:
effects observed, treatment-related
Body weight and weight changes:
effects observed, treatment-related
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Histopathological findings: non-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:
effects observed, treatment-related

Details on results (P0)

P-parents
No test item-related mortality was observed at any dose level.
Rales were observed at a low incidence and on few occasions among the high dosed animals; the high dosed females showed a slightly increased incidence of hunched posture. No nodules or masses were noted.
Body weights were decreased in:
- Males (100 mg/kg bw/d) from the 5th week of treatment onwards, which showed a statistical significant decrease during post mating.
- Females (100 mg/kg bw/d) showed a statistical significant decrease during pregnancy and lactation.
Body weight gain was statistically significantly decreased in:
- Males (100 mg/kg bw/d) during the last half of the pre-mating period.
- Females (100 mg/kg bw/d) during pregnancy.
Body weight and body weight gain for males of the 10 and 30 mg/kg dose group were unaffected.
Food consumption was statistically significantly decreased in:
- Females (100 mg/kg bw/d) during the pre-mating, pregnancy and lactation periods.
Relative food consumption was statistically significantly decreased in:
- Males (100 mg/kg bw/d) during weeks 1, 3 and 5 pre-mating.
- Females (100 mg/kg bw/d) during pre-mating and the first two weeks of pregnancy.
Minor statistically significant differences between controls and 10 or 30 mg/kg bw/d were considered to be of no toxicological relevance.
Details are presented in Table A6.8.2- 2
No test item related macroscopic findings were observed. Findings were considered to be within the range of biological variation of rats of this age and strain.
The number of implantation sites was statistically significant decreased in dams of the 100 mg/kg dose group.
The number of living pups at first litter check (day 1 of lactation) was statistically significantly decreased in the 100 mg/kg dose group.
Total postnatal loss between days 0 and 4 post partum was statistically significantly increased and the viability index was statistically significantly decreased in litters of the 10, 30 and 100 mg/kg dose groups when compared to the control group. However, these findings were considered to be of no toxicological significance.
Total breeding loss between days 5 and 25 post partum was statistically significantly increased in litters of the 30 mg/kg dose group when compared to the control group. This finding was considered to be of no toxicological significance.
Details are presented in Table A6.8.2- 4.


F1-parents
No test item-related mortality was observed at any tested dose level.
Clinical appearance was unaffected when treated up to 100 mg/kg bw/day. An increased incidence in piloerection was noted in animals of the 30 and 100 mg/kg dose groups during the first two weeks of treatment. A slightly increased incidence in hunched posture was noted in males of the 100 mg/kg dose group during the first two weeks of treatment. Because these findings were transient, they were not considered to be of any toxicological significance. One female of the 30 mg/kg bw/d group showed a palpable mass at the chest from week 15 onwards. This single observation was considered to be caused by chance and thus of no toxicological significance.
Body weights of males of the 100 mg/kg dose group were statistically significantly decreased during the pre-mating and mating period.
Females of the 100 mg/kg dose group showed statistically significantly decreased body weights during the first week of the pre-mating period. Because this was a transient effect, it was not considered to be of toxicological significance. Body weight gain was statistically significantly increased in females of the 10 mg/kg dose group during weeks 3–10 of the pre-mating period. Due to the absence of a dose response relationship, this finding was considered to be of no toxicological relevance.
Food consumption was statistically significantly decreased in males of the 100 mg/kg dose group during the pre-mating period and the relative food consumption was also slightly lower. During several weeks of the pre mating period, absolute and relative food consumption were statistically significantly decreased in females treated with 100 mg/kg.
Results are presented in Table A6.8.2- 2.
Macroscopic observations at necropsy did not reveal any alterations that were considered a result of treatment.
Reproduction parameters and breeding data were unaffected by treatment.
Results are presented in Table A6.8.2- 4.

Effect levels (P0)

open allclose all
Dose descriptor:
NOAEL
Remarks:
general parental toxicity
Effect level:
30 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: decreases in body weight, body weight gain and relative food consumption in male and female high dose animals (100 mg/kg bw/day); absolute food consumption was significantly decreased in females only
Dose descriptor:
NOAEL
Remarks:
reproduction
Effect level:
30 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: P-dams of the 100 mg/kg bw/d dose group showed a reduced number of implantation sites at necropsy and living pups at day 1 of lactation. The number of implantations at birth was not significant in dams of the F1-generation.
Key result
Dose descriptor:
NOAEL
Remarks:
general parental toxicity
Effect level:
6 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
other: decreases in body weight, body weight gain and relative food consumption in male and female high dose animals (20 mg a.i./kg bw/day); absolute food consumption was significantly decreased in females only
Key result
Dose descriptor:
NOAEL
Remarks:
reproduction
Effect level:
6 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
other: P-dams of the 20 mg a.i./kg bw/d dose group showed a reduced number of implantation sites at necropsy and living pups at day 1 of lactation. The number of implantations at birth was not significant in dams of the F1-generation.

Target system / organ toxicity (P0)

Critical effects observed:
no

Results: F1 generation

General toxicity (F1)

Clinical signs:
no effects observed
Mortality / viability:
mortality observed, treatment-related
Body weight and weight changes:
no effects observed
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Histopathological findings:
not examined

Details on results (F1)

F1-pups
Development of pups was similar for control and treated groups.
No test-item related macroscopic findings were observed upon necropsy of F1-weanlings.
Mean body weights of pups were similar for control and treated groups.


F2-pups
Development of pups was similar for control and treated groups.
No test-item related macroscopic findings were observed upon necropsy of F2-weanlings.
Mean body weights of pups were similar for control and treated groups.

Effect levels (F1)

Dose descriptor:
NOAEL
Generation:
F1
Effect level:
20 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
other: Clinical signs, body weights and macroscopic examination were unaffected in F1- and F2-pups with maternal treatment up to 20 mg a.i./kg bw/d.

Target system / organ toxicity (F1)

Critical effects observed:
no

Results: F2 generation

Effect levels (F2)

Dose descriptor:
NOAEL
Generation:
F2
Effect level:
20 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
other: Clinical signs, body weights and macroscopic examination were unaffected in F1- and F2-pups with maternal treatment up to 20 mg a.i./kg bw/d.

Target system / organ toxicity (F2)

Critical effects observed:
not specified

Overall reproductive toxicity

Reproductive effects observed:
yes
Lowest effective dose / conc.:
20 mg/kg bw/day (actual dose received)
Treatment related:
yes
Relation to other toxic effects:
reproductive effects as a secondary non-specific consequence of other toxic effects
Dose response relationship:
not specified
Relevant for humans:
not specified

Any other information on results incl. tables

Table A6.8.2-1:Animal assignment for mating.

 

 

Number of animals

 

 

Control

10 mg/kg /daya

30 mg/kg/daya

100 mg/kg/daya

P mating

Male

24

24

24

24

 

Female

24

24

24

24

F1mating

Male

24

23

23

22

 

Female

24

22

23

22

a: based on test substance as manufactured (20% aqueous solution)

 

Table A6.8.2-2:Observed effects on mortality and body weights during the study.

Parameter

 

 

Control

10 mg/kg/d#

30 mg/kg/d#

100 mg/kg/d#

 

 

Generation

Male

Female

Male

Female

Male

Female

Male

Female

Mortality

 

P

 

 

 

1/24

 

 

2/24

1/24

 

 

F1

 

 

 

 

1/23

2/23

4/22

2/22

Body weight

[g]

 

 

 

 

 

 

 

 

 

Pre-mating

Week 1

Week 3

Week 11

 

P

 

260

336

492

 

272

273

n.a.

 

265

344

504

 

270

270

n.a.

 

266

353

491

 

268

271

n.a.

 

267

341

461

 

264

263

n.a.

Post-mating / Pregnancy

Day 0

Day 1

Day 21

 

 

 

n.a.

494

n.a.

 

277

n.a.

450

 

n.a.

513

n.a.

 

271

n.a.

427

 

n.a.

489

n.a.

 

272

n.a.

433

 

n.a.

452*

n.a.

 

261*

n.a.

397**

Lactation

Day 1

Day 14

Day 25

 

 

 

 

324

360

331

 

 

315

355

326

 

 

316

350

318

 

 

295**

331**

303**

Pre-mating

Week 1

Week 5

Week 10

 

F1

 

117

354

494

 

104

232

284

 

109

353

496

 

96

234

299

 

112

347

478

 

99

235

289

 

102*

323*

428**

 

93*

223

279

Pregnancy

Day 0

Day 21

 

 

 

n.a.

n.a.

 

297

453

 

n.a.

n.a.

 

311

478

 

n.a.

n.a.

 

304

469

 

n.a.

n.a.

 

289

432

Body weight gain

[%]

 

 

 

 

 

 

 

 

 

Pre-mating

Week 6

Week 11

 

P

 

58

89

 

n.a.

n.a.

 

59

90

 

n.a.

n.a.

 

57

84

 

n.a.

n.a.

 

50*

73**

 

n.a.

n.a.

Pregnancy

Day 14

Day 21

 

 

 

 

27

63

 

 

25

58

 

 

25

60

 

 

21**

52**

Pre-mating

Week 6

Week 10

[%]

F1

 

235

329

 

141

177

 

263

362

 

172**

223*

 

245

332

 

154

194

 

241

313

 

154

198

Pregnancy

Day 14

Day 21

 

 

 

 

21

53

 

 

20

54

 

 

21

54

 

 

21

50

n.a.: not applicable

*: Dunnett-test statistically significantly different compared to control (p£0.05); ** (p£0.01)
#:in terms of test substance as manufactured (20% aqueous solution)

 

Table A6.8.2-3:Observed effects on food consumption during the study.

Parameter

 

 

Control

10 mg/kg/d#

30 mg/kg/d#

100 mg/kg/d#

 

 

Generation

Male

Female

Male

Female

Male

Female

Male

Female

Food consumption

[g / animal / day]

 

 

 

 

 

 

 

 

 

Pre–mating

Weeks 1/2

Weeks 2/3

 

P

 

28

28

 

22

21

 

29

30

 

21

20

 

29

30

 

21

20

 

28

28

 

19**

18**

Pregnancy

Days 0–7

Days 7–14

Days 14–21

 

 

 

 

26

27

29

 

 

24*

26

27*

 

 

24*

27

28

 

 

22**

23**

25**

Lactation

Days 1–4

Days 7–14

Days 14–21

 

 

 

 

38

59

71

 

 

33

56

67

 

 

36

56

67

 

 

31**

52**

63*

Pre–mating

Weeks 1/2

Weeks 2/3

Weeks 5/6

Weeks 10/11

[g / animal / day]

F1

 

22

28

35

36

 

18

20

25

24

 

21

27

34

35

 

17

20

26

26

 

19*

25

31

31

 

17

20

24

23

 

18*

23*

28*

27**

 

16**

17*

21*

21

Pregnancy

Days 14–21

 

 

 

 

30

 

 

31

 

 

31

 

 

27*

Relative food consumption

[g / kg bw / day]

 

 

 

 

 

 

 

 

 

Pre–mating

Weeks 1/2

Weeks 2/3

 

P

 

93

85

 

79

77

 

94

88

 

76

74

 

94

85

 

78

75

 

89*

83

 

72**

70**

Pregnancy

Days 0–7

Days 7–14

 

 

 

 

82

78

 

 

79*

76

 

 

80

79

 

 

75**

72*

Pre–mating

Weeks 1/2

Weeks 2/3

[g / kg bw / day]

F1

 

118

111

 

121

111

 

117

110

 

120

110

 

109

102

 

118

107

 

117

103

 

116

98*

Pregnancy

Days 14–21

 

 

 

 

67

 

 

64

 

 

65

 

 

63*

n.a.: not applicable
*: Dunnett-test statistically significantly different compared to control (p<
0.05); ** (p<0.01)
#:
based on test substance as manufactured (20% aqueous solution)

 

 

Table A6.8.2-4:Observed effects on reproductive performance of the two generation study.

 

 

 

Control

10 mg/kg/d a

30 mg/kg/d a

100 mg/kg/d a

Parameter

 

Implantations-sites

Mean

P

15.6

14.3

14.7

13.1+

 

F1

14.0

14.9

15.1

12.6

Living pups at 1. litter check

Mean

P

15.3

13.9

14.2

12.9 +

 

F1

13.8

14.1

14.9

13.0

Postnatal loss days 0-4 p.p.

Total

P

11

41##

21#

25##

 

F1

18

38##

12

4##

Breeding loss days 5–25 p.p.

Total

P

1

3

6+

5

 

F1

0

12##

14##

3

Viability index

 

P

97

87.7##

93.8#

91.6##

 

F1

94.1

87.7

96.3

98.4

+: Steel-test significant at 5% level
# /##: Fisher’s exact test significant at 5% (#) or 1% (##) level
p.p. = post partum
a:
based on test substance as manufactured (20% aqueous solution)

 

 

Applicant's summary and conclusion

Conclusions:
In a two generation study, DOPA-Glycinate (20% a.i., as manufactured) was administered to groups of male and female Wistar rats at 0 (control), 10, 30 or 100 mg/kg bw/day in water, corresponding to 2, 6 and 20 mg a.i./kg bw/day, by gavage during the pre-mating and mating period and to females also during gestation and lactation.
No treatment-related mortality occurred during the study. Parental toxicity consisted of clinical signs (only P-animals), effects on body weights and food consumption for P- and F1-animals receiving 100 mg/kg bw/day. Reproductive toxicity consisted of a reduced number of implantation sites at necropsy and a reduced number of living pups on day 1 of lactation for P-animals receiving 100 mg/kg bw/day. Development of the pups was not affected up to the maximum dose of 100 mg/kg bw/day.
Based on the results of this study, the parental and reproductive NOAEL was established at 30 mg/kg bw/day, corresponding to 6 mg a.i./kg bw/day. The developmental NOAEL was established at 100 mg/kg bw/day, corresponding to 20 mg a.i./kg bw/day.
Executive summary:

DOPA-Glycinate (20% a.i., as manufactured) was administered to groups of male and female Wistar rats at 0 (control), 10, 30 or 100 mg/kg bw/day inwater, corresponding to 2, 6 and 20 mg a.i./kg bw/day,by gavage during the pre-mating and mating period and to females also during gestation and lactation. The two generation reproduction study was carried out in compliance with OECD guideline 416 (1983) and also with the recent version OECD 416 (2001), except that the examination of oestrus cycle and sperm parameters were not conducted in the study.

 

Parental toxicity

Parental toxicity was assessed by observing mortality, clinical signs, body weights, food consumption, and macroscopic examination for both generations. Mortality and macroscopic examination were unaffected in parental animals (P) and F1-generation up to 100 mg/kg/day. At 100 mg/kg body weight/day, P-females showed a slightly increased incidence in hunched posture, while P-animals displayed rales at a low incidence and on few occasions. No treatment-related clinical signs were observed in animals of the F1-generation. P- and F1-animals of the 100 mg/kg dose group showed a reduced body weight, body weight gain, food consumption, and relative food consumption during their treatment period. 

Parental NOAEL= 30 mg/kg bw/day, corresponding to 6 mg a.i./kg bw/day based on decreases in body weight, body weight gain and relative food consumption in male and female high dose animals (100 mg/kg bw/day). Absolute food consumption was significantly decreased in females only.

 

Reproductive toxicity

Reproductive toxicity was assessed by observing the number of implantations at birth, mating performance, fertility indices, and breeding data. Mating performance, fertility indices, and breeding data were unaffected in animals of the P- and F1-generations when treated up to 100 mg/kg bw/day. P-dams of the 100 mg/kg bw/day dose group showed a reduced number of implantation sites at necropsy and living pups at day 1 of lactation. The number of implantations at birth was not significant in dams of the F1-generation. 

The increase in postnatal loss in the 10 mg/kg bw/day dose group was due to three litters in which an increased loss was observed. In one of those litters all fifteen pups died spontaneously or were killedin extremisbecause the dam died spontaneously due to delivery difficulties. The increase in postnatal loss in the 30 mg/kg bw/day dose group was mainly due to one litter in which all fourteen pups died on or before day 5post partum. This was probably due to bad health of the dam (hunched posture and body weight loss). The increase in postnatal loss in the 100 mg/kg bw/day dose group was mainly due to two litters in which an increased loss was observed. In one of those litters all nine pups died on or before day 5post partum. This was probably due to poor dam health (hunched posture, piloerection, and reduced body weight). These statistically significant differences were due to single observations, the number of affected litters was similar for control and treatment groups, and no true treatment related effect was observed. There was no dose response associated with these effects. As a consequence of these observations on postnatal loss, the viability index was statistically significantly decreased in litters from all dose groups. These findings are not considered to be toxicologically relevant, they do not display a dose response and as such are not taken into account for determining the reproductive NOAEL. 

Reproductive NOAEL= 30 mg/kg bw/day, corresponding to 6 mg a.i./kg bw/day based on a reduced number (-16% relative to controls) of implantation sites at necropsy of P females. Reduced numbers of living pups on day 1 of lactation for parental animals receiving 10, 30 and 100 mg/kg bw/day were statistically significant but did not follow a dose response and are not taken into account for determining the reproductive NOAEL.

 

Developmental toxicity

Developmental toxicity was assessed by observing clinical signs, body weights and macroscopic examination of the pups during their lactation period. Clinical signs, body weights and macroscopic examination were unaffected in F1- and F2-pups with maternal treatment up to 100 mg/kg body weight/day. 

Developmental NOAEL= 100 mg/kg bw/day, corresponding to 20 mg a.i./kg bw/day. No treatment-related macroscopic pathological effects were observed for any dose tested. 

 

No treatment-related mortality occurred during the study. Parental toxicity consisted of clinical signs (only P-animals), effects on body weights and food consumption for P- and F1-animals receiving 100 mg/kg bw/day. Reproductive toxicity consisted of a reduced number of implantation sites at necropsy and living pups on day 1 of lactation for P-animals receiving 100 mg/kg bw/day.

 

In conclusion, gavage treatment of male and female Wistar rats with DOPA-Glycinate (20% a.i.) at dose levels of 10, 30 or 100 mg/kg bw/day, corresponding to 2, 6 and 20 mg a.i./kg bw/day, over two generations revealed parental, and reproductive toxicity in animals receiving 100 mg/kg bw/day. Development of the pups was not affected up to the maximum dose of 100 mg/kg bw/day. 

 

Based on the results of this study, the parental and reproductive NOAEL was established at 30 mg/kg bw/day, corresponding to 6 mg a.i./kg bw/day. The developmental NOAEL was established at 100 mg/kg bw/day, corresponding to 20 mg a.i./kg bw/day.