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Toxicological information

Developmental toxicity / teratogenicity

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Administrative data

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
Data from experimental study report

Data source

Reference
Reference Type:
review article or handbook
Title:
Teratologic Evaluation of Gentian Violet (CAS No. 548-62-9) in New Zealand White Rabbits
Author:
NTP
Year:
1982
Bibliographic source:
National Center for Toxicological Research, TER82080 (study conducted in 1981-82)

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
other: As mentioned below
Principles of method if other than guideline:
Teratogenicity of Gentian Violet (CAS No. 548-62-9) in New Zealand White Rabbits was evaluated following maternal exposure
GLP compliance:
not specified
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid
Details on test material:
- Name of test material (as cited in study report):Gentian violet
- IUPAC name: N-(4-{bis[4-(dimethylamino)phenyl]methylene}cyclohexa-2,5-dien-1-ylidene)-N-methylmethanaminium chloride
- Molecular formula : C25H30N3.Cl
Molecular weight : 407.986 g/mol
- Smiles notation: C(\c1ccc(N(C)C)cc1)(c1ccc(N(C)C)cc1)=C1\C=C\C(=[N+](/C)C)C=C1.[ClH-]
- InChl:1S/C25H30N3.ClH/c1-26(2)22-13-7-19(8-14-22)25(20-9-15-23(16-10-20)27(3)4)21-11-17-24(18-12-21)28(5)6;/h7-18H,1-6H3;1H/q+1;/p-1
- Substance type: Organic
- Physical state: Solid
Specific details on test material used for the study:
Name of test material (as cited in study report): Gentian violet
- IUPAC name: N-(4-{bis[4-(dimethylamino)phenyl]methylene}cyclohexa-2,5-dien-1-ylidene)-N-methylmethanaminium chloride
- Molecular formula : C25H30N3.Cl
Molecular weight : 407.986 g/mol
- Smiles notation: C(\c1ccc(N(C)C)cc1)(c1ccc(N(C)C)cc1)=C1\C=C\C(=[N+](/C)C)C=C1.[ClH-]
- InChl:1S/C25H30N3.ClH/c1-26(2)22-13-7-19(8-14-22)25(20-9-15-23(16-10-20)27(3)4)21-11-17-24(18-12-21)28(5)6;/h7-18H,1-6H3;1H/q+1;/p-1
- Substance type: Organic
- Physical state: Solid

Test animals

Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
Details on test animal & Environmental conditions

TEST ANIMALS
- Source:Dutchland Laboratory Animals, Inc. (Denver, PA)
- Age at study initiation:approximately six months 14 days.
- Weight at study initiation:Female : 3.5-4.6 kg
- Housing:stainless steel cages with mesh flooring Minimum cage dimensions were 18" x 24" x 18" and maximum cage dimensions were 24" x 24" x
20".
- Diet (e.g. ad libitum):Purina Certified Rabbit Chow ad libitum
- Water (e.g. ad libitum):deionized/filtered water ad libitum
- Acclimation period:14days

ENVIRONMENTAL CONDITIONS
- Temperature (°C):16.66-22.22°C
- Humidity (%):57.13 ± 2.86%
- Air changes (per hr):No data avaliable
- Photoperiod (hrs dark / hrs light):12 hr

IN-LIFE DATES: From: To:


Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
Gentian violet was dissolved in distilled water in concentrations of 0, 0.5, 1.0 and 2.0 mg/mlmg/ml for the conventional teratology study.
Dosing solutions were stored at 5°C except at the time of dosing when they were warmed to room temperature, and stirred continuously with a magnetic stirrer
Analytical verification of doses or concentrations:
yes
Remarks:
by spectrophotometry
Details on analytical verification of doses or concentrations:
yes,by spectrophotometry
Details on mating procedure:
Artificially inseminated rabbits
Duration of treatment / exposure:
13 days i.e. Gestational days 6 through 19
Frequency of treatment:
Daily
Duration of test:
30 days
Doses / concentrations
Remarks:
0.0, 0.5, 1.0 or 2.0 mg/kg/day
No. of animals per sex per dose:
Total:133
0.0mg/kg : 30
0.5mg/kg: 32
1.0mg/kg: 31
2.0mg/kg: 40
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: The doses were selected based on the pilot study

Examinations

Maternal examinations:
CAGE SIDE OBSERVATIONS:No data available

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: on gestational days 0, 6-19

BODY WEIGHT: Yes
- Time schedule for examinations:Dams were weighed on gestational days 0, 6-19 (prior to daily dosing) and 30 (immediatelyprior to sacrifice)

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): No data
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data
- Time schedule for examinations:

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day # :sacrifice on gestational day 30,
- Organs examined: liver weight, gravid uterine weight and status of uterine implantation sites (i.e., implantation sites, resorptions, dead fetuses, live fetuses).

OTHER:
Ovaries and uterine content:
No data available The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
- Other:
Fetal examinations:
- External examinations: Yes:Live fetuses were dissected from the uterus and evaluated for live litter size, body weights, sex ratios and gross morphological abnormalities.

- Soft tissue examinations: Yes: All live fetuses were examined for visceral malformations employing the Staples' fresh tissue dissection method.

- Skeletal examinations: Yes: All fetal carcasses were cleared and stained with Alizarin Red S and examined for skeletal malformations.

- Head examinations: Yes: half per litter :Half of the fetuses were decapitated prior to dissection and theheads were fixed in Bouin's solution for free hand sectioning and examination (Wilson's Technique).
Statistics:
ANOVA was performed on selected response variables 30 employing General Linear Hodel (GLH) procedures for the Statistical Analysis System (SAS) Library .Prior to GLM analysis, the arcsine-square root transformation was performed on all maternal or litter-derived percentage data (Snedecor and Cochran, 1967), and Bartlett's test for homogeneity of variance (alpha level =0.001) (Winer, 1962) was performed on all data to be analyzed by GLM.The statistical tests that were performed on each parameter are described in Appendix II (protocol), Amendment II, Table 2 and Amendment1. Two dependent variables (i.e. average fetal body weight per litter, and percent malformed fetuses per litter), were analyzed in a three-way(dose x replicate x sex) ANOVA design with sex as a repeated measure within each litter. When either of these variables showed a significant (p<0.05) interaction between sex and dose as tested against the general error term, the data for that variable were examined separately for eachsex in a two-way (dose x replicate) design. When either variable showed a nonsignificant (p>0.05) dose x sex interaction.
Indices:
No data available
Historical control data:
Yes. -- Historical data on 1142 New Zealand White rabbit control matings have been summarized across 75 teratology studies from pharmaceutical companies or biological research organizations by the Middle Atlantic Reproduction and Teratology Association
(Woo and Hoar, 1982)

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Clinical signs were taken daily during the dosing period (gd 6 through 19), and at necropsy (gd 30). The following symptoms were seen in does in a dose-related manner: wheezing, diarrhea, congestion, wet nose, dyspnea, lacrimation, and anorexia and cyanosis (the latter two in those which died).

Dermal irritation (if dermal study):
not specified
Mortality:
mortality observed, treatment-related
Description (incidence):
Maternal mortality followed a clear dose-response trend with seven deaths (out of 31 retained in the study, 22.6%) in the 2.0 mg/kg/day dose group, four deaths (out of 26 retained in the study, 15.4%) in the 1.0 mg/k/day dose group and two deaths (out of 27 retained in the study, 7.4%) in the 0.5 mg/kg/day dose group. No deaths occurred in the control group (out of 27 does retained in the study).
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
A number of weight related parameters also indicated maternal toxicity. There was a significant downward trend for maternal body weight on gd 19 (at the end of the dosing period), and maternal weight gain (gestation period and treatment period).Significant pairwise comparisons were present only for maternal weight gain, where all gentian violet-exposed groups were significantly lower than controls for this parameter for the gestation period and treatment period.
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
There were no treatment- related effects on gravid uterine weight, liver weight or relative liver weight.
Gross pathological findings:
not specified
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
not specified
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Details on results:
Clinical signs and mortality
Clinical signs: in a dose-related manner, included wheezing, diarrhea, congestion, wet nose, dyspnea, lacrimation, and anorexia and cyanosis (the latter two in those does which died).

Mortality: The maternal mortality in the present study was 22.6% (7/31 does) in the 2 mg/kg bw dose group, 15.4% (4/26) in the 1 mg/kg bw dose group, 7.4% (2/27) in the 0.5 mg/kg bw dose group and 0.0 (0/27) in the control group.

Dermal irritation: No data

Body weight and weight gain: There was a significant downward trend for maternal body weight on gd 19

Food consumption and compound intake: No data

Food efficiency: No data

Water consumption and compound intake: No data

Opthalmoscopic examination: No data

Haematology: No data available

Clinical chemistry: No data available

Urinanalysis: No data available

Neurobehaviour: No data

Organ weights: There were no treatment- related effects on gravid uterine weight, liver weight or relative liver weight.

Gross pathology: No data available
Histopathology: No data available

Maternal developmental toxicity

Number of abortions:
no effects observed
Description (incidence and severity):
There were no treatment related effects on the number or percent of dead conceptuses per litter. There was a significant upward trend in the percentage of non-live (dead plus resorbed) and affected (non-live plus malformed) conceptuses per liter but no significant pairwise comparisons
Pre- and post-implantation loss:
no effects observed
Description (incidence and severity):
Examination of reproductive parameters indicated a significant increase in the number of implantation sites per litter in all compound-exposed groups versus controls. Percent resorptions per litter and number of litters with resorptions exhibited a dose-related upward trend but no significant pairwise comparisons
Total litter losses by resorption:
no effects observed
Description (incidence and severity):
Percent resorptions per litter and number of litters with resorptions exhibited a dose-related upward trend but no significant pairwise comparisons
Early or late resorptions:
not specified
Dead fetuses:
effects observed, treatment-related
Description (incidence and severity):
For litters with one or more live fetuses (i.e “live litters") the number of fetuses (male and/or female) did not differ among dose groups.
Changes in pregnancy duration:
not specified
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): not specified
Changes in number of pregnant:
not specified
Other effects:
not specified

Effect levels (maternal animals)

Dose descriptor:
NOEL
Effect level:
2 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
clinical signs
mortality
body weight and weight gain
maternal abnormalities

Results (fetuses)

Fetal body weight changes:
no effects observed
Description (incidence and severity):
Average fetal body weight per litter exhibited a significant dose-related downward trend with values from all gentian violet exposed groups significantly lower than from controls. When separated by sex, only the female foetuses exhibited the significant pairwise comparisons
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): no effects observed
Field "Description (incidence and severity)" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.DescriptionIncidenceAndSeverityFetalPupBodyWeightChanges): Average fetal body weight per litter exhibited a significant dose-related downward trend with values from all gentian violet exposed groups significantly lower than from controls. When separated by sex, only the female foetuses exhibited the significant pairwise comparisons
Reduction in number of live offspring:
not specified
Changes in sex ratio:
no effects observed
Description (incidence and severity):
For litters with one or more live fetuses (i.e “live litters") the number of fetuses (male and/or female) did not differ among dose groups
Changes in litter size and weights:
not specified
Changes in postnatal survival:
not specified
External malformations:
no effects observed
Description (incidence and severity):
There were no significant dose-related effects on incidence of gross malformations per litter, nor in the number (or percent) of males, females or foetuses malformed per litter, nor in the number or percent of litters with malformed fetuses
Skeletal malformations:
no effects observed
Description (incidence and severity):
There were no significant dose-related effects on incidence of skeletal malformations per litter, nor in the number (or percent) of males, females or foetuses malformed per litter, nor in the number or percent of litters with malformed fetuses
Visceral malformations:
effects observed, treatment-related
Description (incidence and severity):
There were no significant dose-related effects on incidence of visceral malformations per litter, nor in the number (or percent) of males, females or foetuses malformed per litter, nor in the number or percent of litters with malformed fetuses
Other effects:
not specified
Details on embryotoxic / teratogenic effects:
Fetal body weight changes: When separated by sex, only female fetal body weight per litter exhibited a significant downward trend and pairwise comparisons. When separated by sex, only female fetal body weight per litter exhibited a significant downward trend and pairwise comparisons.
Reduction in no of live offspring: The number of fetuses (male and/or female) per litter did not differ among dose groups
Changes in sex ratio: no effects observed
Fetal/pup body weight changes: Average fetal body weight per litter exhibited a significant downward trend with all dose group values significantly lower than controls.
Changes in litter size and weight: no effects observed
Changes in postnatal survival: no effects observed
External malformation: no effects observed
Skeletal malformation: no effects observed
Visceral malformation: no effects observed
Other effect: No data available

Effect levels (fetuses)

Dose descriptor:
NOAEL
Effect level:
2 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
changes in sex ratio
changes in litter size and weights
changes in postnatal survival
external malformations
skeletal malformations
visceral malformations

Overall developmental toxicity

Developmental effects observed:
no
Lowest effective dose / conc.:
2 mg/kg bw/day (nominal)
Treatment related:
no
Relation to maternal toxicity:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified

Any other information on results incl. tables


Assignment of CD Rats in Replicate and Dose Groups in the Teratological Evaluation of Gentian Violet

 

Gentian Violet (mg/kg/day, po)

Replicate

0.0

2.5

5

10

I

14

13

14

14

II

18

17

17

18

Total Treated

32

30

31

32

Summary of Analysis of Teratological Defects Observed in CD Rat Fetuses Following Maternal Exposure to Various Dose Levels of Gentian Violet by Oral Gavage on Gestational Days 6 Through 15

 

Gentian Violet (mg/kg/day, po)

 

0

2.5

5

10

 fetuses examineda

264

299

264

235

 litter examinedb

25

26

24

22

Gross malformations per litterc

0

0

0

0

Visceral malformations

 

 

 

0

Foetusesc

0

2

1

5f

Litterd

0

1

1

4

Skeletal malformations

 

 

 

 

Foetusesc

1

2

5

5

Litterd

1

2

3

5

Fetuses malformed per litterc

0.04

±0.04

0.15

±0.09

0.25

±0.14

0.45**

±0.14

Percent fetuses malformed per litterc

0.31

±0.31

1.50

±0.86

1.91

±1.02

6.70**

±2.70

No. d fetuses malformed

1

4

6

10

Percent fetuses malformedd

0.4

1.3

2.3

4.3

No. litters with malformationse

1

3

4

8

Percent litters with malformationse

4.0

11.5

18.7

36.4

Males malformed per litterc

0.04h

±0.04

0.08h

±0.06

0.09h

±0.06

0.29h

±0.12

Females malformed per litterc

0.0

1.41

0.98

2.56

1.53

8.30

5.00

Percent males malformed per litterc

0.52

±0.52

1.20

±0.88

1.24

±0.86

4.54

±1.95

Percent females malformed per litterC

0.0

1.41

±0.98

2.56

±1.53

8.30

±5.00

aOnly live fetuses were examined for malformations.

blncludes only litters with live feluses.

cfetuses with one or more malformations.

dLitters with one or more malformed fetuses.

eReported as mean + S.E.H. for all live litters.

fOne litter ofhead(7) was lost.

gOne fetus was lost during staining process.

hOne litter had no male fetuses:

iOne litter had no female fetuses.

jTwolitter. had no female fetuse

§p<0.05 Jonckheere's Test.

§§p<0.01Jonckheere's Test.

*p<0.05 Kruskal-Wallis.

**p<0.01 Mann-Whitney U.

***p<0.05 Fisher's Exact Test.

Specific Teratological Defects Observed Following Maternal Exposure to Various Dose Levels of Gentian Violet by Oral Gavage on Gestational Days 6 Through 15

 

Gentian Violet (mg/kg/day, po)

 

00

2.5

5.0

10

Total fetuses examinedb

264

299

264

235

Total litters examinedc

25

26

24

22

GROSS MALFORMATIONS f

0

0

0

0

VISCERAL MALFORMATIONSg

 

 

 

 

Hydronephrosis: left

 

1

 

 

right

 

 

 

2

Hydroureter: bilateral

 

 

 

1

right

 

 

 

2

left

 

2

1

1

Extra liver lobe

 

 

 

1

SKELETAL Malformations

 

 

 

 

Lumbar centra misaligned

 

 

1

 

Thnracic centra off center

 

1

1

 

Short rib

1

1

3

5

Missing ribf

 

 

1

 

VARIATIONSf,g,h

 

 

 

 

Hematoma: jaw

 

1

 

 

neck

1

 

 

 

lower limb

1

 

 

 

back

 

1

 

 

No. innominate

 

1

1

1

wavy rib

3

 

 

 

Misaligned sternebrae

2

2

2

3

Extra ossification sites -

 

6

 

 

spinal cord and tail

 

 

 

 

Doubled centra

 

5

3

1

Clubbed limb w/o bone change

 

 

1

 

Bilateral papilla 1/4 normalsize or less

1

 

 

1

Left papilla 1/4 normalsize or less

 

 

 

1

Right papilla 1/4 normalsize or less

2

 

1

1

Very soft tissue - kidney

 

1

 

 

Incomplete ossification

 

1

1

 

Distended ureter(s)

21

18

8

15

aA single fetus may have more than one malformation or variation,

bAll live fetuses were examined for gross malformations.

cAll live fetuses were examined for visceral malformations of the trunk. Fifty percent of the heads were examined for soft tissue malformations.

dFifty percent decapitated carcasses and50%fetal preparations with heads intact were examined for skeletal malformations.

eThirteenth rib was two-thirds or less normal length, unilaterally or bilaterally.

fone fetus had both twelfth and thirteenth ribs missing.

 

Reproductive Parameters after Exposure of Pregnant CD Rats to Various Dose Levels of Gentian Violet by Oral Gavage on Gestational Days 6 Through 15

 

 

Gentian Violet (mg/kg/day, po)

 

0.0

2.5

5.0

10

ALL LITTERSa

 

 

 

 

(Pregnant Dams)

25

26

24

25

Implantation sites per 1itterb

11.48

±0.70

12.35

±0.53

11.50

±0.86

11.52

±0.77

Resorptions per litterb

0.92

±0.38

0.85

±0.51

0.50

±0.17

2.12

±0.93

Percent Resorptions per litterb

9.3

±3.8

5.6

±3.1

5.4

±2.1

17.5

±6.6

No. Litters with resorptions

8

7

8

11

Percent litters with resorptions

32.0

26.9

33.3

44.0f

Dead per litterb

0

0

0

0

Percent dead per litterb

0

0

0

0

No. Litters with dead

0

0

0

0

Percent Litters with dead

0

0

0

0

0Non-live per litterb,c

0.92

±0.38

0.85

±0.51

0.50

±0.17

 

2.12

±0.93

Percent non-live per litterb, c

9.3

±3.8

5.8

±3.1

 

5.4

±2.1

17.5

±6.6

No. litters with non-live

8

7

8

11

Percent litters with non-live

32.0

26.9

33.3

44.0

Affected per 1itterb ,d

0.96

±0.39

1.00

±0.51

0.75

±0.21

 

2.52

±0.91

Percent affected per litterb

9.5

±3.8

7.2

±3.2

7.2

±2.3

 

22.3

±6.7

No. litters with affected

8

9

11

17

Percent litters with affected

32.0

34.6

45.8

68.0

LIVE LITTERSe

 

 

 

 

(No. litters with live fetuses)

25

26

24

22

Live fetuses per litterb

10.56

±0.85

11.50

±0.59

11.00

±0.89

10.68

±0.82

Males per litterb

5.56

±0.57

6.04

±0.53

5.25

±0.49

 

5.18

±0.52

Percent males per litterb

53.1

±4.2

52.0

±3.9

 

50.2

±4.5

48.2

±3.9

Average fetal body weight (g) per litter

3.686

±0.077

3.642

±0.059

3.486

±0.072

3.606

±0.090

Average male fetal body weight (g) per litter

3.741

±0.078

3.688

±0.067

3.587

±0.080

3.767

±0.061

Average female fetal body weight (g) per litter

3.560

±0.069

3.553

±0.073

3.422

±0.073

3.510

±0.088

 

aincludes all dams pregnant at sacrifice; litter size = no. implantation sites per dam.

bReported as mean! S.E.M.

cNon-live =dead plus resorbed.

dAffected =non-live plus malformed.

eincludes only dams with live fetuses; litter size =no. live fetuses per dam.

fTwo dams had all resorptions.

gOne litter had no males.

hOne litter had no females.

iOne litter had no females.

ip,0.05 Jonckhecre'. Test.

'p<0.05Fisher exact test

Maternal Parameters after Exposure to Pregnant CD Rats to Various Dose Levels of Gentian Violet by Oral Gavage on Gestational Days 6 Through 15

 

 

Gentian Violet (mg/kg/day, po)

 

0.0

2.5

5

10

SUBJECTS (Dams)

 

 

 

 

Total treated

32

30

31

32

Removed

0

0

0

0

Deaths

0

0

0

0

Non-pregnant at sacrifice

7

4

7

4

Pregnant (%) at sacrifice

25 (78.1)

26(86.7)

24(77.4)

25(86.2)

MATERNAL BODY WEIGHT(g)

 

 

 

 

Gestational day 0a

228.55

±3.11

232.87

±3.51

227.90

±2.97

234.17

±4.14

Gestational day 6a

250.20

±3.51

 

251.83

±3.96

248.47

±3.10

255.58

±3.73

Gestational day 11

272.25

±3.52

272.08

±3.86

262.70

±3.81

250.72

±5.69

Gestational day 15

290.12

±4.26

288.99

±3.97

277.00

±4.39

268.39

±6.93

Gestational day 20

345.00

±6.03

347.83

±6.52

334.89

±6.53

319.01

±9.21

Maternal Weight Gain (g)

 

 

 

 

Gestation period

116.45

±4.38

114.96

±5.85

106.98

±5.34

84.84

±9.35

Treatment period

39.92

±1.36

37.15

±2.16

27.29

±3.27

12.81

±2.13

Absolute weight gain

54.08

±3.36

47.74

±4.38

46.01

±3.30

30.09

±6.13

Gravid Uterine Weight (g)a

62.37

±4.38

67.22

±3.23

60.98

±4.64

54.75

±5.49

Maternal Liver Weight (g)a

14.15

±0.26

14.40

±0.29

13.74

±0.35

13.46

±0.47

Relative Maternal Liver Weight

4.10

±0.08

4.10

±0.09

4.11

±0.07

4.18

±0.08


aincludes alldams pregnant at sacrifice;

bweight gain during gestation minus gravid uterine weight.

CThree dams hadall resorptions.

dOne liver weight had not recorded.

eOne liver weight was incorrectly recorded.

ip<0.05Jonckheeres̔ Test.

iip<0.01Jonckheeres̔ Test.

iiip0.001Jonckheeres̔ Test.

tp<0.05 Kruskal-wallis.

ttp<0.01 Kruskal-Wallis.

tttp<0.001 Kruskal-walli&

 p<0.01Mann-whintlleyU

 p<0.01Mann-whintlleyU.

 

 

 

 

 

 

Applicant's summary and conclusion

Conclusions:
NOAEL for maternal toxicity and fetal toxicity was considered to be 2mg/kg bw /day i.e highest tested dose. When New Zealand white rabbits were treated with Gentian violet (548-62-9) orally.
Executive summary:

The teratogenicity study of Gentian violet (548-62-9) was performed in femaleNew Zealand white rabbits. Total 133 animals used for study in which 30 female animals in control group. The doses were selected base upon preliminary toxicity study. All the animals provided with food and water adlibitum.Gentian violet was dissolved in distilled water in concentrations of 0, 0.5,1. 0 and 2.0 mg/ml for the conventional teratology study. Dosing solutions were stored at 5°C except at the time of dosing when they were warmed to room temperature, and stirred continuously with a magnetic stirrer and analytical verification done using spectrophotometry.

 

Artificially inseminated rabbits were used .Timed-pregnant CD rats were given gentian violet at dose levels of 0.0, 0.5, 1.0 or 2.0 mg/kg/day in distilled water. Dams were weighed on gestational days 0, 6-19 (prior to daily dosing) and 30 (immediately prior to sacrifice), and were also observed for clinical signs of toxicity. At sacrifice on gestational day 30, dams were evaluated for body weight, liver weight, gravid uterine weight and status of uterine implantation sites (i.e., implantation sites, resorptions, dead fetuses, live fetuses). Live fetuses were dissected from the uterus and evaluated for live litter size, body weights, sex ratios and gross morphological abnormalities.All live fetuses were examined for visceral malformations employing the Staples' fresh tissue dissection method. Half of the fetuses were decapitated immediately after dissection and the heads were fixed in Bouin's solution for free hand sectioning and examination (Wilson's Technique). All fetal carcasses were cleared and stained with Alizarin Red S and examined for skeletal malformations.

 

The maternal mortality in the present study was 22.6% (7/31 does) in the 2mg/kg bw dose group, 15.4% (4/26) in the 1mg/kg bw dose group, 7.4% (2/27) in the0. 5mg/kg bw dose group and 0.0 (0/27) in the control group. A significant trend was seen toward reduction in maternal body weight on gestational day 19 (end of dosing), and in maternal weight gain (gestational period and treatment period). For maternal weight gain, all GV-exposed groups were significantly lower than for controls for both treatment and gestation period. Clinical signs, seen in a dose-related manner, included wheezing, diarrhea, congestion, wet nose, dyspnea, lacrimation, and anorexia and cyanosis (the latter two in those does which died). All GV-exposed groups exhibited a significant increase in the number of implantation sites per litter versus controls. Percentage of resorptions per litter and number of litters with resorptions, as well as the percentage of non-live (dead plus resorbed) and affected (non- live plus malformed) per litter exhibited a dose-related upward trend, but no significant pairwise comparisons. For live litters, the number of fetuses (male and/or female) per litter did not differ among dose groups. Average fetal body weight per litter exhibited a significant downward trend with all dose group values significantly lower than controls. When separated by sex, only female fetal body weight per litter exhibited a significant downward trend and pairwise comparisons.

There were no significant dose-related effects on the incidence of gross, visceral or skeletal malformations per litter, nor in the number (or percent) of fetuses, males or females, malformed per litter nor in the number or percent of litters with malformed fetuses. Examination of malformation incidence by category indicated no malformations unique to or with a higher incidence in any of the GV-exposed groups relative to control. No evidence of teratogenicity of gentian violet was seen when administered by gavage to pregnant NZW rabbits during organogenesis at doses which produced evidence of maternal and fetal mortality and toxicity. Hence NOAEL for maternal toxicity and fetal toxicity was considered to be 2mg/kg bw /day i.e highest tested dose. When New Zealand white rabbits were treated with Gentian violet (548-62-9) orally.