Reaction mass of aluminium fluoride and chromium trifluoride and nickel difluoride

Toxicity to reproduction

Administrative data

Endpoint:

three-generation reproductive toxicity

Remarks:

based on test type (migrated information)

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

1960-1962 (in-life phase)

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study in accordance with generally accepted methods, but limited level of parameters examined and data reported

Data source

Reference

Reference Type:

publication

Title:

Unnamed

Year:

1976

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 416 (Two-Generation Reproduction Toxicity Study)

Deviations:

yes

Remarks:

lower number of animals used and parameters examined

GLP compliance:

no

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Nickel sulfate hexahydrate (NiS04.6H20)

IUPAC Name:

Nickel sulfate hexahydrate (NiS04.6H20)

Details on test material:

Nickel sulfate hexahydrate fines containing 22.3% nickel by analysis (theoretically 22.32%) served as the test material in this study.

Test animals

Species:

rat

Strain:

other: Wistar derived

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Albino Farms, Red Bank, New Jersey, USA
- Age at study initiation: (P) 4 wks
- Weight at study initiation: (P) 56-65 g
- Fasting period before study: No data
- Housing: individually caged
- Use of restrainers for preventing ingestion (if dermal): yes/no
- Diet: ad libitum, Radlston Purina, St. Louis, Missouri, USA
- Water: ad libitum
- Acclimation period: No data

ENVIRONMENTAL CONDITIONS
No details provided

IN-LIFE DATES: From: 1960 To: 1962

Administration / exposure

Route of administration:

oral: feed

Vehicle:

other: Radlston Purina feed

Details on exposure:

DIET PREPARATION
- Rate of preparation of diet (frequency): weekly
- Mixing appropriate amounts with (Type of food): Finely ground laboratory chow served as the basic diet. Stock aqueous solutions of nickel sulfate hexahydrate were prepared in appropriate concentrations so that the addition of 100 mL for each 6 kg of diet resulted in the desired nickel content, respectively. Additions of nickel solutions were thoroughly blended into the diet by mixing in a rotary-type blender.
- Storage temperature of food: No data

Details on mating procedure:

- M/F ratio per cage: 1/1
- Length of cohabitation: 7 days
- Proof of pregnancy: No data
- Further matings after two unsuccessful attempts: No
- After successful mating each pregnant female was caged: individually
- Any other deviations from standard protocol: On the 20th mating day all males were removed.

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

11 weeks prior to mating, during mating, gestation, and lactation

Frequency of treatment:

Daily

Details on study schedule:

Litters containing more than 10 offsprings were randomly reduced to 10 on day 5. All surviving F/1a siblings were sacrificed and autopsied at weaning. Approximately 10 days after weaning of F/1a litters, F/0 parent generation rats were remated for F/1b litters. Procedure and observations recorded were the same as those described for F/1a litters.

For the F/2 generation 30 male and 30 female F/1b offsprings from each diet level were continued on their respective parents' diet for 11 weeks at which time 20 of each sex within each group were mated and the same procedure followed as with the F/0 generation through production and weaning of F/2a and F/2b litters.

For the F/3 generation, the same procedure as with the previous generations was followed through the production of F/3a and F/3b litters. All matings in each generation were made with rats from different litters.

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 250, 500 and 1000 ppm
Basis:
nominal in diet
equivalent to 0, 22.5, 45 and 90 mg/kg bw nickel, respectively

No. of animals per sex per dose:

30

Control animals:

yes, plain diet

Details on study design:

No further details provided

Positive control:

Not included

Examinations

Parental animals: Observations and examinations:

Weekly body weight records were obtained, except during mating through weaning of litters.
Records were maintained of mating and number of pregnancies.

Oestrous cyclicity (parental animals):

Not included

Sperm parameters (parental animals):

Not included

Litter observations:

Records were maintained of litters cast (alive and dead), pups in litter at 1, 5, and 21 days (weaning), and total weight of the litter at weaning.

Postmortem examinations (parental animals):

Following weaning of F/1b litters, surviving F/0 rats were sacrificed and autopsied.

Postmortem examinations (offspring):

At weaning of F/2b rats, F/1b parents were sacrificed and autopsied.
Histopathologic examinations were performed on 10 male and 10 female F/3b weanlings from each diet level. Absolute and relative organ-to-body weight ratios for heart, spleen, kidneys, liver, and testes were analysed statistically. Tissues preserved in formalin for histopathologic study were in addition to the above: lung, urinary bladder, stomach, small and large intestine, skeletal muscle, brain, skin, bone marrow, pituitary, thyroid, adrenal, pancreas, and gonad.

Statistics:

No data

Reproductive indices:

Fertility (F.l.) = (pregnancies/matings) x 100
Gestation (G. I.) = (litters cast/pregnancies) x 100

Offspring viability indices:

Viability (V.I.) = (live pups at day 5/live pups born) x 100
Lactation (L.I.) = (weaned/live pups-discards on day 5) x 100

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

not specified

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

decreased bodyweight at 1000 ppm (90 mg/kg)

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

decreased bodyweight at 1000 ppm (90 mg/kg)

Organ weight findings including organ / body weight ratios:

not examined

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:

no effects observed

Details on results (P0)

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
Body weights for parent generation rats on 250 and 500 ppm diets (22.5 and 45 mg/kg nickel, resp.), before mating and at weaning of respective litters, were not adversely affected, but rats on 1000 ppm (90 mg/kg nickel) exhibited slightly lower body weights. The average decrease in body weight did not exceed 8% for females and 13% for males.

OTHER FINDINGS (PARENTAL ANIMALS)
On fertility and gestation indices, no adverse effects were noted at any of the dietary levels of nickel.

Effect levels (P0)

Dose descriptor:

NOAEL

Remarks:

reproductive toxicity

Effect level:

1 000 ppm (nominal)

Based on:

test mat.

Sex:

male/female

Results: F1 generation

General toxicity (F1)

Clinical signs:

not specified

Mortality / viability:

mortality observed, treatment-related

Description (incidence and severity):

higher incidence of stillborn in the first generation

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

decreased at 1000 ppm (90 mg/kg nickel)

Sexual maturation:

no effects observed

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

no effects observed

Histopathological findings:

no effects observed

Details on results (F1)

VIABILITY (OFFSPRING)
Data on the number of pups born dead showed higher incidence of stillborn in the first generation at all levels of nickel, not observed to any extent in subsequent generations. The number of siblings (alive and dead) cast per litter averaged 10.3, 10.6, 9.8, and 9.0 for 0, 250, 500, and 1000 ppm (0, 22.5, 45, and 90 mg/kg nickel, resp.) diets, respectively. The number of siblings weaned per litter were progressively fewer with increasing dietary level of nickel, averaging 8.1, 7.2, 6.8, and 6.4 for 0, 250, 500, and 1000 ppm (0, 22.5, 45, and 90 mg/kg nickel, resp.) diets, respectively.

BODY WEIGHT (OFFSPRING)
On average weaning body weight, a clear-cut adverse effect is only apparent in weanlings of females on 1000 ppm (90 mg/kg nickel) diet, averaging 73% of control. However, offsprings maintained on 1000 ppm (90 mg/kg nickel) diet from weaning to mating of succeeding generations recovered considerably from this deficit, averaging 92% of controls.

OTHER FINDINGS (OFFSPRING)
On viability and lactation indices, no adverse effects were noted at any of the dietary levels of nickel.

Effect levels (F1)

Dose descriptor:

LOAEL

Generation:

F1

Effect level:

250 ppm (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Higher incidence of stillborn

Overall reproductive toxicity

Reproductive effects observed:

not specified

Any other information on results incl. tables

Summary table of reproduction data for rats fed with nickel sulfate hexahydrate through 3 generations:

Generation		Dose (ppm)	Number of rats			Total number of pups					Weanlings mean body weight (g)	Indices (%)*
			Mated **	Pregnant ***	Whelped^§	Born alive	Born dead	Alive on day 5^§§	Discarded^§§§	Weaned		F.I.	G.I.	V.I.	L.I.
F0 =>	F1a	0	20(1)	15(1)	14	113	5	113(10)	2	89	37.5	79	93	100	87
		250	20(1)	11	11(1)	72	17	69	5	60	40.1	58	100	96	94
		500	19	14	14(1)	96	13	95	5	72	34.4	74	100	99	80
		1000	20	12	12(1)	93	16	91	5	83	27.8	60	100	98	97
	F1b	0	17	14	14	143	3	139	0(8)	137	33.8	82	100	97	99
		250	19	16	16	164	6	150	0(12)	134	31.6	84	100	91	89
		500	19	14	14(3)	109	27	106	0(9)	98	28.1	74	100	97	93
		1000	20(1)	15	15	93	31	77	0(3)	67	25.1	79	100	83	87
F1b =>	F2a	0	20	15	15	142	0	138	13	115	35.9	75	100	97	92
		250	20	18	18(1)	188	2	181	30	119	31.1	90	100	96	79
		500	20	17	17	171	0	141	10	129	34.6	85	100	82	98
		1000	20	16(2)	14	127	6	120	8	96	25.9	80	88	94	86
	F2b	0	20	14	14(1)	149	1	146	30	116	37.9	70	100	98	100
		250	20	17	17(1)	181	16	177	39	130	35.7	85	100	98	94
		500	20	16(1)	15	166	0	160	22	131	36.9	80	94	96	95
		1000	18	11	11(1)	109	3	103	10	90	26.9	61	100	94	97
F2b =>	F3a	0	20	18(1)	17	180	5	175	20	125	36.7	90	94	97	81
		250	20	19	19	207	4	201	21	121	34.5	95	100	97	67
		500	20	19(1)	18(1)	184	5	172	20	102	35.6	95	95	93	67
		1000	20	20(1)	19	168	7	162(10)	6	107	280.	100	95	96	73
	F3b	0	18	18	18	216	0	206	38	158	41.9	100	100	95	94
		250	20	18	18	204	2	198	32	161	41.4	90	100	97	97
		500	18	16	16(1)	142	6	138	22	102	41.2	89	100	97	88
		1000	17	17	17	139	7	138	13	114	29.7	100	100	99	91

* Indices: F.I., G.I., V.I., and L.I. refer to Fertility, Gestation, Viability, and Lactation Index, respectively.

** Numbers within parentheses refers to non-pregnant dams found dead during mating period, not included in calculation of F.I.

*** Within parentheses, dams found dead and autopsied to confirm pregnancies.

^§ Numbers within parentheses refers to litters born dead

^§§ Numbers within parentheses refers to number of siblings sacrificed after day 5 and before weaning due to death of dam, not used in calculation of L.I.

^§§§ To reduce litter size to 10 on day 5; and within parentheses are totals in excess of 10/litter not discarded as planned, due to oversight, and carried to weaning.

Applicant's summary and conclusion

Conclusions:

Based on the only occurrence of slightly decreased body weight at 1000 ppm (90 mg/kg nickel) in the parental generation, the NOAEL for parental reproductive toxicity can be set at 90 mg nickel/kg/day.
Based on increased lethality in F1 pups, the low dose of 250 ppm (22.5 mg/kg nickel) can be considered as a LOAEL.

Executive summary:

In a 3-generation study, Wistar-derived rats were fed with 0, 250, 500, or 1000 ppm of nickel sulfate hexahydrate (equivalent to 0, 22.5, 45, or 90 mg/kg of nickel, respectively).

In the parental generation, body weights for rats on 250 and 500 ppm diets (22.5 and 45 mg/kg nickel, resp.), before mating and at weaning of respective litters, were not adversely affected, but rats on 1000 ppm (90 mg/kg nickel) exhibited slightly lower body weights. The average decrease in body weight did not exceed 8% for females and 13% for males. On fertility and gestation indices, no adverse effects were noted at any of the dietary levels of nickel.

A higher incidence of stillborn was observed only in the first generation at all dietary levels of nickel and decreased body weights of weanings on 1000 ppm diet in all generations. On viability and lactation indices, no adverse effects were noted at any of the dietary levels of nickel.