2-(4-amino-2-nitroanilino)ethanol

Developmental toxicity / teratogenicity

Administrative data

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 16 July 2004 to 1 July 2005

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

GLP Study. Eight instead of twenty females were used in this study. Complete analysis was not performed as required in the OECD Guideline.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2005

Report date:

2005

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Deviations:

yes

Remarks:

8 females per condition were used instead of 20 required. Complete analysis was not performed

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

2-(4-amino-2-nitroanilino)ethanol

EC Number:

220-701-7

EC Name:

2-(4-amino-2-nitroanilino)ethanol

Cas Number:

2871-01-4

Molecular formula:

C8H11N3O3

IUPAC Name:

2-(4-amino-2-nitroanilino)ethanol

Test material form:

solid: particulate/powder

Remarks:

Crystalline

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: supplied by Procter & Gample, Batch No. L-35939
- Expiration date of the lot/batch: 8 July 2005
- Purity test date:8 July 2003

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: stored at room temperature
- Stability under test conditions: the test item was considered as stable refrigerated or at room temperature
- Solubility and stability of the test substance in the solvent/vehicle: The concentration verification samples were acceptable (test item in vahicle analysis), as they did not vary from the theorical value by more than 15%. For the homogeneity analysis, the overall relative standard deviation from all sampling locations for each respective concentration was within 5%.

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
Formulations were prepared at least every two weeks based on established stability of the dose formulations. Prepared formulations were stored refrigerated; the actual temperature range in the storage refrigerator from the day of the first preparation to the last day of dosage administration was 0°C to 8°C. Prepared formulations were stirred continuously using a magnetic stir bar and stir plate during sample collection and dosage.

Test animals

Species:

rat

Strain:

Crj: CD(SD)

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories
- Age at study initiation: 62 Days (females) ; 69 Days (males)
- Weight at study initiation: 228-254g (Females) ; 504-740g (Males, at cohabitation)
- Fasting period before study: Not specified
- Housing: rats were housed in stainless steel, wire-bottomed cages, except during the cohabitation period. During this period, rats were housed in the male rat's cage.
- Diet (e.g. ad libitum): Certified Rodent Diet #5002 (PMI Nutrition International, USA) ad libitum
- Water (e.g. ad libitum): Local water was provided by an automatic watering access system and/or individual water bottles attached to the cage.
- Acclimation period: 5 Days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18.8 to 23.1°C
- Humidity (%): 44.6 to 73.1% relative humidity
- Air changes (per hr): a minimum of ten changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours light / 12 hours dark

IN-LIFE DATES: From: 10 June 2004 (Females) ; 27 March 2004 (Males) To:9 September 2004 (Females)

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

polyethylene glycol

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
The test article was appropriately weighed onto a piece of weigh paper or into an quantitatively sized, labeled, pre calibrated beaker. A small amount of vehicle was added to the beaker and stirred using a spatula. Once the test substance was completely wetted, vehicle was added to the final volume. The dosing solution was continuously stirred using a magneic stirrer for homogeneity.

VEHICLE
- Justification for use and choice of vehicle (if other than water): No justification
- Concentration in vehicle: 0, 10, 40, 100, 200 mg/mL
- Amount of vehicle (if gavage): 5 mL/kg
- Batch No. A10622, A15618

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

A portion of the study was performed in order to determine the concentration and homogeneity of the test item in prepared formulation samples. The samples were quantified on a HPLC system utilizing a reverve-phase column with gradient elution and an ultraviolet detector at 254 nm). The mean concentration results for all samples analysed varied from 87.3 to 99.5% of the respective theorical value. For the homogeneity analyses, the overall relative standard deviation from all samples was within 5%.

Details on mating procedure:

- Impregnation procedure: cohoused
- If cohoused:
- M/F ratio per cage: 1/1
- Length of cohabitation: 5 Days
- After 5 days of unsuccessful pairing replacement of first male by another male with proven fertility.
- Further matings after two unsuccessful attempts: no
- Verification of same strain and source of both sexes: Not specified
- Proof of pregnancy: vaginal smear and/or copulatory plug referred to as day 0 of pregnancy

Duration of treatment / exposure:

Day 6 to Day 20 of pregnancy

Frequency of treatment:

Once daily

Duration of test:

Until the Day 20 of pregnancy

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

0 mg/kg bw/day (nominal)

Doses / concentrations 2

Dose / conc.:

50 mg/kg bw/day (nominal)

Doses / concentrations 3

Dose / conc.:

200 mg/kg bw/day (nominal)

Doses / concentrations 4

Dose / conc.:

500 mg/kg bw/day (nominal)

Doses / concentrations 5

Dose / conc.:

1 000 mg/kg bw/day (nominal)

No. of animals per sex per dose:

8 Females were used per condition

Control animals:

yes

yes, concurrent vehicle

Details on study design:

- Dose selection rationale:
Dosages were selected by the Sponsor on the basis of existing data on this test material. The highest dosage level was selected to induce some overt developmental and/or maternal toxicity (clinical signs or a decrease in body weight) but not death or severe suffering. At least one intermediate dosage level was selected to produce minimal observable toxicity. The lowest dosage level should not produce any evidence of either maternal or developmental toxicity. A descending sequence of dosage levels was selected with the purpose of demonstrating any
dosage-related response and to establish a no-observed-adverse-effect level (NOAEL).

- Rationale for animal assignment (if not random):
Upon arrival, rats were assigned to individual housing on the basis of computer-generated random units. Healthy, mated female rats were assigned to five dosage groups (Groups I through V), eight rats per dosage group, by using a computer-generated (weight-ordered) randomization procedure based on body weights recorded on DG 0.

Examinations

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes
Rats were observed for viability at least twice each day of the study and for clinical observations and general appearance weekly during the acclimation period and on DG 0.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily

BODY WEIGHT: Yes
- Time schedule for examinations: Body weights were recorded weekly during the acclimation period, on DG 0 and daily during the dosage and post-dosage periods.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes


POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day #21
- Organs examined: Caesarean-sectioned and a gross necropsy of the thoracic, abdominal and pelvic viscera was performed. The number and distribution of corpora lutea were recorded. The uterus of each rat was excised, weighed and examined for pregnancy, number and distribution of implantation sites, live and dead fetuses and early and late resorptions

Ovaries and uterine content:

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

Fetal examinations:

Fetuses were examined for sex and for gross external alterations

Statistics:

Averages and percentages were calculated. Litter values were used where appropriate. Additional procedures and/or analyses may be performed if deemed appropriate

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

Clinical observations considered to be related to the test substance included purple skin; purple perioral substance; dark purple, purple, orange or red urine; and purple or orange fur, which were observed in all of the groups treated with the test substance. These color changes are attributable to the presence of the test substance, a dye, and are not considered to be adverse. All other clinical signs were not considered to be related to the test substance because: 1) a similar number of rats were affected with the sign in the control group; 2) only one to three rats were affected with the sign in any dosage group; and 3) the number of affected rats in each dosage group did not occur in a dosage-dependent manner.
These clinical signs included soft or liquid feces; excess salivation; localized alopecia on the limbs or back; scab; sparse hair; incisors missing/broken; ungroomed coat; and/or red perivaginal substance.

Dermal irritation (if dermal study):

not examined

Mortality:

no mortality observed

Description (incidence):

All rats survived to scheduled sacrifice.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

Mean body weights were comparable among the dosage groups through DG 21; gravid uterine weights and corrected maternal body weights (DG 21 body weight minus the gravid uterine weight) were comparable among the dosage groups.
Mean body weight gains were reduced in the 200, 500 and 1000 mg/kg/day dosage groups on DGs 6 to 9, following the initiation of dosage, as well as on DGs 15 to 18, in comparison with the respective control group values; however, the reductions were not dosage dependent. Mean body weight gains were comparable among the dosage groups for the entire dosage period (calculated as DGs 6 to 21) and the gestation period (DGs 0 to 21), in comparison with the respective control group values. Values remained comparable among the dosage groups during these same intervals when these values were corrected for gravid uterine weights.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

Mean absolute and relative feed consumption values were comparable among the dosage groups for the entire dosage period (calculated as DGs 6 to 21) and the gestation period (DGs 0 to 21), in comparison with the respective control group values.

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

not examined

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

effects observed, non-treatment-related

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

not examined

Histopathological findings: neoplastic:

not examined

Other effects:

not examined

Maternal developmental toxicity

Number of abortions:

no effects observed

Pre- and post-implantation loss:

no effects observed

Total litter losses by resorption:

no effects observed

Early or late resorptions:

no effects observed

Dead fetuses:

no effects observed

Changes in pregnancy duration:

not examined

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 examined

Changes in number of pregnant:

not examined

Other effects:

not examined

Details on maternal toxic effects:

Pregnancy occurred in all eight rats in the 0 (Vehicle), 50, 200, 500 and 1000 mg/kg/day dosage groups. No Caesarean-sectioning or litter parameters were affected by dosages of the test substance as high as 1000 mg/kg/day. The litter averages for corpora lutea, implantations, litter sizes, live fetuses, fetal body weights, percent resorbed conceptuses, and percent live male fetuses were comparable among the five dosage groups. No dam had a litter consisting of only resorbed conceptuses, and there were no dead fetuses. All placentae appeared normal. The average numbers of early resorptions in the 200, 500 and 1000 mg/kg/day dosage groups were slightly increased, and the average number of late resorptions in the 1000 mg/kg/day was slightly increased, but the values were within the range of historical control data for this Testing Facility and not considered treatment related.

Effect levels (maternal animals)

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Basis for effect level:

other: overall observations

Results (fetuses)

Fetal body weight changes:

no effects observed

Description (incidence and severity):

Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): not examined

Reduction in number of live offspring:

not examined

Changes in sex ratio:

no effects observed

Changes in litter size and weights:

no effects observed

Changes in postnatal survival:

not examined

External malformations:

no effects observed

Skeletal malformations:

not examined

Visceral malformations:

not examined

Other effects:

not examined

Details on embryotoxic / teratogenic effects:

Totals of 125, 113, 117, 114 and 120 fetuses from 8, 8, 8, 8 and 8 litters in the 0 (Vehicle), 50, 200, 500 and 1000 mg/kg/day dosage groups, respectively, were examined for gross external alterations. There were no fetal gross external alterations.

Effect levels (fetuses)

Key result

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

changes in sex ratio

external malformations

Overall developmental toxicity

Developmental effects observed:

no

Any other information on results incl. tables

Table 1 :Clinical Signs–Summary

GROUP	I			II			III			IV			V
DOSAGE (MG/KG/DAY)a	0			50			200			500			1000
MAXIMUM POSSIBLE INCIDENCE		128/	8		128/	8		128/	8		128/	8		128/		8
MORTALITY		0			0			0			0			0
PURPLE SKIN		0/	0		94/	8		122/	8		128/	8		127/		8
PURPLE PERIORAL SUBSTANCE		0/	0		46/	7		95/	8		120/	8		107/		8
DARK PURPLE OR PURPLE OR ORANGE OR RED URINE b		0/	0		27/	5		25/	7		49/	8		62/		8
PURPLE OR ORANGE: FUR		0/	0		11/	2		47/	6		101/	8		119/		8
SOFT OR LIQUID FECES		2/	2		4/	3		0/	0		3/	2		2/		1
EXCESS SALIVATION		0/	0		0/	0		0/	0		0/	0		1/		1
LOCALIZED ALOPECIA:TOTAL		12/	1		16/	1		0/	0		23/	2		0/		0
LIMB(S)		12/	1		16/	1		0/	0		9/	1		0/		0
BACK		0/	0		0/	0		0/	0		14/	1		0/		0
RIGHT FORELIMB: SHOULDER,SCAB		0/	0		0/	0		0/	0		5/	1		0/		0
SPARSE HAIR		0/	0		0/	0		14/	1		0/	0		0/		0
INCISOR(S):MISSING/BROKEN		0/	0		1/	1		0/	0		0/	0		0/		0
UNGROOMED COAT		3/	2		7/	1		0/	0		0/	0		0/		0
RED PERIVAGINAL SUBSTANCE		1/	1		0/	0		0/	0		0/	0		0/		0

 

 

MAXIMUM POSSIBLE INCIDENCE = (DAYS x RATS)/NUMBER OF RATS EXAMINED PER GROUP ON DAYS 6 THROUGH 20 OF GESTATION.

N/N = TOTAL NUMBER OF OBSERVATIONS/NUMBER OF RATS WITH OBSERVATION.

a.   Dosage occurred on days 6 through 20 ofgestation.

b.   Observation was located on the head, both ears, neck, chest, both forelimbs, both forepaws, lower midline, rightflank, left flank, back, right side of back, left side of back, left and right axilla, bothhindlimbs, bothhindpaws, tail and/or wholebody.

 

TABLE 2(PAGE 1): NECROPSY OBSERVATIONS - SUMMARY

 

--------------------------------------------------- GROUP   I            II                                  III         IV          V

DOSAGE(MG/KG/DAY)a               0           50          200          5001000

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RATS EXAMINED b	N	8	8	8	8	8
MORTALITY	N	0	0	0	0	0
APPEARED NORMAL	N	6	8	8	8	7
UMBILICAL HERNIA: SKIN SURROUNDING UMBILICUS	N	2	0	0	0	1

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a.   Dosage occurred on days 6 through 20 ofgestation.

b.   Refer to the individual clinical observations table (Table 10) for external observations confirmed atnecropsy.

 

TABLE 3: MATERNAL BODY WEIGHTS AND GRAVID UTERINE WEIGHTS -SUMMARY

 

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GROUP	I	II	III	IV	V
DOSAGE (MG/KG/DAY)a	0	50	200	500	1000

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RATS TESTED			N	8		8			8			8			8
PREGNANT			N	8		8			8			8			8
MATERNAL BODY WEIGHT		(G)
DAY	0	MEAN±S.D.		240.5	±	7.7	241.0	±	8.2	240.5	±	6.8	243.0	±	8.4	240.4	±	7.4
DAY	6	MEAN±S.D.		274.2	±	8.7	277.5	±	6.4	274.4	±	8.4	283.5	±	14.9	273.1	±	8.8
DAY	7	MEAN±S.D.		277.8	±	13.5	277.4	±	5.9	271.6	±	12.0	280.1	±	14.3	273.9	±	9.6
DAY	8	MEAN±S.D.		280.1	±	12.1	281.2	±	7.5	275.5	±	9.6	289.2	±	18.0	278.0	±	7.3
DAY	9	MEAN±S.D.		285.8	±	12.9	288.2	±	9.4	281.9	±	11.9	291.8	±	15.2	282.9	±	10.1
DAY	10	MEAN±S.D.		289.2	±	13.6	294.0	±	8.7	288.2	±	10.7	298.9	±	16.7	286.2	±	9.7
DAY	11	MEAN±S.D.		295.4	±	15.1	299.1	±	11.4	294.5	±	14.4	304.8	±	16.8	297.1	±	11.8
DAY	12	MEAN±S.D.		300.2	±	15.6	306.2	±	12.3	297.8	±	13.8	311.0	±	16.7	302.0	±	12.3
DAY	13	MEAN±S.D.		303.2	±	13.3	307.9	±	11.1	306.4	±	15.2	320.1	±	19.1	310.0	±	12.7
DAY	14	MEAN±S.D.		310.8	±	13.6	313.6	±	10.0	311.6	±	15.3	326.9	±	19.8	313.8	±	13.9
DAY	15	MEAN±S.D.		320.0	±	13.2	322.1	±	11.1	319.2	±	15.8	337.2	±	24.2	323.6	±	15.8
DAY	16	MEAN±S.D.		332.2	±	15.0	333.8	±	12.4	332.2	±	15.6	346.0	±	19.9	333.8	±	15.3
DAY	17	MEAN±S.D.		348.0	±	16.8	348.9	±	15.6	343.0	±	18.6	360.8	±	22.6	346.1	±	15.0
DAY	18	MEAN±S.D.		364.9	±	17.3	366.2	±	17.2	358.9	±	18.6	376.8	±	25.6	363.2	±	17.0
DAY	19	MEAN±S.D.		382.2	±	17.7	380.5	±	19.4	377.0	±	17.9	396.6	±	27.9	380.2	±	19.8

 

DAY = DAY OF GESTATION

a. Dosageoccurred on days 6 through 20 of gestation.

 

TABLE 3:MATERNAL BODY WEIGHTS AND GRAVID UTERINE WEIGHTS - SUMMARY

 

----------------------------------------------- GROUP    I            II                               III         IV          V

DOSAGE(MG/KG/DAY)a            0           50          200          500  1000

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RATSTESTED         N         8            8           8            8                    8

PREGNANT            N         8            8           8            8      8

 

MATERNAL BODY WEIGHT (G)

 

DAY20       MEAN±S.D.401.4±18.4399.6±19.9 393.4±22.3412.9±27.3              392.6 ±19.5

DAY21       MEAN±S.D.424.1±21.3424.5±26.5 418.4±21.1435.4±29.7          419.0±18.7 GRAVIDUTERINE

WEIGHT(G)    MEAN±S.D.115.7±7.6 108.4±30.0 110.5±12.3108.6±10.1              113.9 ±11.1

 

DAY21Cb    MEAN±S.D.308.4±15.4316.1±14.2 307.9±13.6326.8±22.8              305.1 ±14.1

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DAY = DAY OF GESTATION

a.   Dosage occurred on days 6 through 20 ofgestation.

b.   21C = Corrected maternal body weight (day 21 of gestation body weight minus the gravid uterineweight).

 

TABLE 4:CAESAREAN-SECTIONING OBSERVATIONS - SUMMARY

 

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GROUP	I	II	III	IV	V
DOSAGE (MG/KG/DAY)a	0	50	200	500	1000

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RATS TESTED	N	8	8		8		8		8
PREGNANT	N(%)	8(100.0)	8(100.0)		8(100.0)		8(100.0)		8(100.0)
RATS PREGNANT AND CAESAREAN-SECTIONED
ON DAY 21OFGESTATION                  N		8		8		8		8		8
CORPORALUTEA                 MEAN±S.D.		17.1 ±	1.2	17.8 ±	3.0	16.2 ±	2.0	16.4 ±	1.4	17.5 ±	1.3
IMPLANTATIONS                 MEAN±S.D.		16.2 ±	1.0	14.5 ±	3.7	15.5 ±	1.4	15.2 ±	1.7	16.1 ±	1.1
LITTERSIZES                 MEAN±S.D.		15.6 ±	0.5	14.1 ±	3.5	14.6 ±	2.0	14.2 ±	1.7	15.0 ±	1.6
LIVEFETUSES               N		125		113		117		114		120
MEAN±S.D.		15.6 ±	0.5	14.1 ±	3.5	14.6 ±	2.0	14.2 ±	1.7	15.0 ±	1.6
DEADFETUSES               N		0		0		0		0		0
RESORPTIONS                 MEAN±S.D.		0.6 ±	0.7	0.4 ±	0.7	0.9 ±	0.8	1.0 ±	1.4	1.1 ±	1.4
EARLYRESORPTIONS               N		5		3		7		8		7
MEAN±S.D.		0.6 ±	0.7	0.4 ±	0.7	0.9 ±	0.8	1.0 ±	1.4	0.9 ±	1.4
LATERESORPTIONS               N		0		0		0		0		2
MEAN±S.D.		0.0 ±	0.0	0.0 ±	0.0	0.0 ±	0.0	0.0 ±	0.0	0.2 ±	0.5
DAMS WITH ANYRESORPTIONS	N(%)	4( 50.0)		2( 25.0)		5( 62.5)		4( 50.0)		5( 62.5)
DAMS WITH ALLCONCEPTUSES RESORBED	N(%)	0(0.0)		0(0.0)		0(0.0)		0(0.0)		0(0.0)
DAMS WITH VIABLE FETUSES	N(%)	8(100.0)		8(100.0)		8(100.0)		8(100.0)		8(100.0)
PLACENTAE APPEAREDNORMAL	N(%)	8(100.0)		8(100.0)		8(100.0)		8(100.0)		8(100.0)

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a. Dosageoccurred on days 6 through 20 of gestation.

 

 

 

TABLE 5: LITTER OBSERVATIONS (CAESAREAN-DELIVERED FETUSES) - SUMMARY

 

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GROUP	I		II		III		IV		V
DOSAGE(MG/KG/DAY)a            0           50          200         500       1000 ------------------------------------------------------------------------------------------------------------------------------------
LITTERS WITH ONE OR MORE LIVE FETUSES	N	8		8		8		8		8
IMPLANTATIONS	MEAN±S.D.	16.2 ±	1.0	14.5 ±	3.7	15.5 ±	1.4	15.2 ±	1.7	16.1 ±	1.1
LIVE FETUSES	N	125		113		117		114		120
	MEAN±S.D.	15.6 ±	0.5	14.1 ±	3.5	14.6 ±	2.0	14.2 ±	1.7	15.0 ±	1.6
LIVE MALE FETUSES	N	58		63		58		53		61
% LIVE MALE FETUSES/LITTER	MEAN±S.D.	46.2 ±	10.8	55.2 ±	11.7	49.8 ±	17.0	46.5 ±	15.5	51.3 ±	13.9
LIVE FETAL BODY WEIGHTS
(GRAMS)/LITTER	MEAN±S.D.	5.49 ±	0.24	5.52 ±	0.72	5.66 ±	0.41	5.65 ±	0.26	5.47 ±	0.23
MALE FETUSES	MEAN±S.D.	5.63 ±	0.22	5.64 ±	0.66	5.80 ±	0.42	5.82 ±	0.24	5.61 ±	0.27
FEMALE FETUSES	MEAN±S.D.	5.39 ±	0.24	5.38 ±	0.76	5.53 ±	0.40	5.50 ±	0.26	5.32 ±	0.31

% RESORBED

CONCEPTUSES/LITTER         MEAN±S.D.   3.6±4.2    2.3±4.3    5.9±5.8               6.2±     8.8  6.9±  8.4

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a. Dosageoccurred on days 6 through 20 of gestation.

 

 

TABLE 6: FETAL GROSS EXTERNAL ALTERATIONS - SUMMARY

 

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GROUP	I	II	III	IV	V
DOSAGE (MG/KG/DAY)a	0	50	200	500	1000

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LITTERS EVALUATED	N	8	8	8	8	8
FETUSES EVALUATED	N	125	113	117	114	120
LIVE	N	125	113	117	114	120
DEAD	N	0	0	0	0	0

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NO FETAL ALTERATIONS WERE IDENTIFIED AT GROSS EXTERNAL EXAMINATION

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a. Dosageoccurred on days 6 through 20 of gestation.

 

 

 

 

 

 

Applicant's summary and conclusion

Conclusions:

Under the experimental codition of the study, the Test Article HC Red No. 3 induced no significant differences in body weight between treated and control animals. No significant differences the litter sizes were observed between treated and control animals. No significant differences in the number of live or resorbed foetuses were observed between treated and control animals. No differences in foetal abnormalities were observed between treated and control groups. The NOAEL for materno and embryo-toxicity was considered to be 1000 mg/kg/day. Based on the results of this range-finding study, the doses selected for the main study were 0, 50, 200 and 1000 mg/kg bw/d.

Executive summary:

The purpose of this GLP compliant study was to provide information for the selection of dosages to be used in the developmental toxicity study.

Forty presumed pregnant Crl:CD rats, eight per group, were randomly assigned to five dosage groups. Formulations of the test substance, HC Red 3, and/or the vehicle, 100% polyethylene glycol 400 (PEG 400), were administered orally once daily to rats on days 6 through 20 of gestation (DGs 6 through 20) at dosages of 0, 50, 200, 500 and 1000 mg/kg/day. The dosage volume was 5 mL/kg. Viabilities, clinical observations, body weights and feed consumption values were recorded. All rats were sacrificed on DG 21. The gravid uterus was weighed and examined for the number and distribution of corpora lutea, implantation sites and uterine contents. Fetuses were weighed and examined for gross external alterations and sex. Caesarean-sectioning and subsequent fetal observations were conducted without knowledge of dosage group.

Under the experimental codition of the study, the Test Article HC Red No. 3 induced no significant differences in body weight  between treated and control animals. No significant differences the litter sizes were observed between treated and control animals. No significant differences in the number of live or resorbed foetuses were observed between treated and control animals. No differences in foetal abnormalities were observed between treated and control groups. The NOAEL for materno and embryo-toxicity was considered to be 1000 mg/kg/day. Based on the results of this range-finding study, the doses selected for the main study were 0, 50, 200 and 1000 mg/kg bw/d.