Ammonium undecafluorohexanoate

Toxicity to reproduction

Administrative data

Endpoint:

one-generation reproductive toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

12 Apr 2006 - 13 Jul 2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Reference

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

12 Apr 2006 - 13 Jul 2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)

Version / remarks:

adopted 21 Sept 1998

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.3100 (90-Day Oral Toxicity in Rodents)

Version / remarks:

(1998)

Deviations:

no

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

other: Crl:CD(SD)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Inc., Raleigh, North Carolina, U.S.A.
- Females nulliparous and non-pregnant: not specified
- Age at study initiation: 6 - 8 weeks
- Housing: individually in stainless steel, wire-mesh cages
- Diet: PMI® Nutrition International, LLC Certified Rodent LabDiet® 5002, ad libitum
- Water: tap water, ad libitum


DETAILS OF FOOD AND WATER QUALITY:
Water samples are analyzed for total bacterial counts, and the presence of coliforms, lead, and other contaminants. Certified animal feed is used, guaranteed by the manufacturer to meet specified nutritional requirements and not to exceed stated maximum concentrations of key contaminants,
including specified heavy metals, aflatoxin, chlorinated hydrocarbons, and organophosphates. The presence of these contaminants below the maximum concentration stated by the manufacturer would not be expected to impact the integrity of the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 - 26
- Humidity (%): 30 - 70
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:
Dose concentrations in vehicle: 4, 20 and 100 mg/mL
Dose volume: 5.0 mL/kg bw

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Concentration verification and stability of dosing formulations was measured from 4 different collecting time points during the study: near the beginning, near the middle (approximately test day 45) and near the end of the exposure phase of the study. Concurrent with dosing formulation analyses, recovery of the test substance from spiked NANOpure® water was tested at the low level (approximately 4 mg/mL), the middle level (approximately 20 mg/mL), and the high level (approximately 100 mg/mL) to confirm the analytical method.
Data from the analysis of the samples during the study indicate that the test substance was at the targeted concentrations, mixed uniformly, and stable under the conditions of the study. Test substance was not found in the control group samples.

Duration of treatment / exposure:

90 days

Frequency of treatment:

once daily, 7 days/weeks

Dose / conc.:

20 mg/kg bw/day (actual dose received)

Dose / conc.:

100 mg/kg bw/day (actual dose received)

Dose / conc.:

500 mg/kg bw/day (actual dose received)

No. of animals per sex per dose:

10 (main study and 90-day recovery group, respectively; and 30-day recovery group for control and high-dose only)

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Dose level selection was based on the results of a preliminary dose range-finding study, were animals were orally exposed to 0, 500 and 1000 mg/kg bw/day for 2 weeks. Test substance-related effects on hematology (decreased red cell mass, hemoglobin and hematocrit; increased reticulocytes and platelets) and clinical chemistry (increased AST and ALT) occurred in male rats dosed with 500 or 1000 mg/kg bw/day. In addition, an increase in absolut and relative liver weights was observed in animals receiving 500 or 1000 mg/kg bw/day test substance. Animals dosed with 500 and 1000 mg/kg bw/day test substance exhibited mildly decreased cholesterol and triglyceride levels and decreases in total protein due to decreases in globulin. Therefore, 500 mg/kg bw/day was selected as the highest dose level for the main study.
- Rationale for selecting satellite groups: 30- and 90-day recovery groups were included to investigate the reversibility of any observed toxicology effects.

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily for mortality/moribundity and daily for general condition; additionally 2 h post dosing ± 30 min for acute clinical signs of toxicity
- Cage side observations included: general condition and clinical signs, mortality/viability

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Day 0 and weekly thereafter
- Observations included: evaluation of fur, skin, eyes, mucous membranes, occurrence of secretions and excretions, autonomic nervous system activity (lacrimation, piloerection, and unusual respiratory pattern), changes in gait, posture, response to handling, presence of clonic, tonic, stereotypical, or bizarre behavior

BODY WEIGHT: Yes
- Time schedule for examinations: once/ week; additionally rats designated for neurobehavioral evaluations, undergoing functional observational battery and motor activity assessments, were weighed on the days of those observations

FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/day: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No

FOOD EFFICIENCY:
- Mean daily food efficiacy (g body weight gain/g food consumed) was calculated from food consumption and body weight data.

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No

OPHTHALMOSCOPIC EXAMINATION: Yes
- Dose groups that were examined: all rats (Day 10) and all suviving rats of subchronic toxicity and 30-day recovery group (Day 81)

HAEMATOLOGY: Yes
- Time schedule for collection of blood: all animals: Day 43/44 and 92/93; recovery animals: 120/121 and 183/184 (males/females)
- Anaesthetic used for blood collection: Yes (CO2)
- Animals fasted: Yes, for at least 15 h
- How many animals: all animals
- Parameters checked: red blood cell count, red cell distribution width, hemoglobin, absolute reticulocyte count, hematocrit, platelet count, mean corpuscular (cell) volume, white blood cell count, mean corpuscular (cell) hemoglobin, differential white blood cell count, mean corpuscular (cell) hemoglobin concentration, microscopic blood smear examination (not for recovery group on Day 183/184), prothrombin time, activated partial thromboplastin time, coagulation (not for recovery group on Day 183/184)

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: all animals: Day 43/44 and 92/93; recovery animals: 120/121 and 183/184 (males/females)
- Animals fasted: Yes, for at least 15 h
- How many animals: all animals
- Parameters checked: aspartate aminotransferase glucose, alanine aminotransferase, total protein, sorbitol dehydrogenase, albumin, alkaline phosphatase, globulin, total bilirubin, calcium, urea nitrogen, inorganic phosphorus, creatinine, sodium, cholesterol, potassium, triglycerides, chloride, fluoride (analysed on EDTA plasma)

URINALYSIS: Yes
- Time schedule for collection of urine: for all animals on Day 43/44 and 92/93; recovery animals: 120/121 and 183/184 (males/females)
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes, for at least 15 h
- Parameters checked: volume, appearance (quality, color, clarity), osmolality, pH, glucose, ketone, bilirubin, blood, urobilinogen, fluoride, protein, microscopic urine sediment examination

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: prior to dosing and during week 13 (over a 2-day period)
- Dose groups that were examined: all animals: The pretest evaluation included spare rats,and rats for a 30-day recovery subset. Based on the results of the week 13 evaluation, the 30-day recovery functional observational battery and motor activity assessment was not necessary.
- Battery of functions tested: Functional observational battery (FOB; Sensory and motor function assessments by evaluating grip strength, responses to approach/touch, sharp auditory stimulus, and tail pinch, pupillary constriction, fore- and hindlimb grip strength); motor activity (MA)

IMMUNOLOGY: No

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
Main group animals were sacrificed for gross examinations on Days 92/93 (males/females) and recovery animals on additional 30 or 90 days later

ORGAN WEIGHTS: Yes
The following tissues were weight from main groups and 30-day recovery groups: liver, kidneys, adrenal glands, thymus, brain, spleen, heart, thyroid gland (weighed after fixation), ovaries (with oviducts), uterus (with cervix), testes and epididymides. Group mean absolute organ weight values and organ weight ratios (% body weight and % brain weight) were calculated. For 90-day recovery groups only gross lesions were examined and liver, spleen, and thyroid gland were weighed.

HISTOPATHOLOGY: Yes
The following tissues were collected from the animals of the main groups and 30-day recovery groups: liver, esophagus, stomach, duodenum, jejunum, ileum, cecum, colon, rectum, salivary glands, pancreas, kidneys, urinary bladder, lungs, trachea, nose (4 sections), larynx, pharynx, heart, aorta, spleen, thymus, mandibular lymph node, mesenteric lymph node, bone marrow (femur and sternum), Peyer´s patches (intestine), pituitary gland, thyroid gland, parathyroid glands, adrenal glands, brain (cerebrum, midbrain, cerebellum, medulla/pons), spinal cord (cervical, mid-thoracic, lumbar), sciatic nerve, skeletal muscle, femur/knee joint, sternum, testes, epididymides, prostate, seminal vesicle, ovaries (incl. oviduct), uterus (incl. cervix), mammary glands, skin and eyes (incl. retina and optic nerve).
Testes, epididymides, and eyes were fixed in modified Davidson’s solution. All other tissues were fixed in 10% neutral buffered formalin. Processed tissues were embedded in paraffin, sectioned approximately 5-6 µm thick, stained with hematoxylin and eosin (H&E), and examined microscopically by a veterinary pathologist.
Target organs (liver, spleen, thyroid, bone marrow, and nose) identified and collected in the main study were evaluated microscopically in the 30-day recovery animals; target organs identified in the 30-day recovery groups were evaluated in the 90-day recovery groups. In addition, gross lesions identified in 30- or 90-day recovery animals were evaluated microscopically.

Statistics:

The statistic significance was calculated at p < 0.05. Separate analyses were performed on the data collected for each sex.
Body weight, body weight gain, food consumption, food efficiency, clinical pathology and organ weight were tested preliminary with Levene’s test for homogeneity and Shapiro-Wilk test for normality. If the Shapiro-Wilk test was not significant but Levene's test was significant, a robust version of Dunnett's test was used. If the Shapiro-Wilk test was significant, a Kruskal-Wallis test was followed by Dunn’s test.
If the preliminary test was not significant, One-way analysis of variance followed by Dunnett's test was used. If the preliminary test was significant Kruskal-Wallis test followed by Dunn's test was used.
Survival and incidence of FOB was tested by Cochran-Armitage test for trend. If the incidence was not significant, but a significant lack of fit occurred, then Fisher’s Exact test with a Bonferroni correction was used.
For motor activity (Test day and 10-minute interval were used as repeated-measure factors) and grip strength repeated measures analysis of variance followed by linear contrasts was used when the preliminary test was not significant. If the preliminary test was significant sequential application of the Jonckheere-Terpstra trend test was performed.

Clinical signs:

no effects observed

Description (incidence and severity):

No test substance-related clinical signs were observed. The clinical signs noted did not exhibit a dose response and no statistically significant increase in incidence was observed.

Mortality:

mortality observed, non-treatment-related

Description (incidence):

There were no test substance-related deaths. Two female rats died prior to their scheduled sacrifice. 1/10 females at 100 mg/kg bw/day was sacrificed in extremis on test day 50. The likely cause of death in this animal was cranial trauma, based on the gross finding of hemorrhage in the dorsal skull area. 1/10 females at 500 mg/kg bw/day was found dead on test day 5. The likely cause of death in this animal was renal papillary necrosis. Treatment-related kidney lesions were not observed in this study; therefore, the kidney change and associated early death in this animal were not considered to be treatment-related. All other rats survived until their respective scheduled terminal sacrifices.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

No test substance-related effects on mean body weight or mean body weight gain were observed in any female group after 90 days of dosing. Statistically significant decreases (95% to 91.3% of control group) in mean body weight were observed in main group and recovery group male rats in the high-dose group (500 mg/kg bw/day) beginning on study day 42 and remaining statistically decreased through study day 105 (2 weeks after ceasing dosing), compared with the concurrent control group. After 30 and 90 days of recovery, no statistical differences were observed in the mean body weight for any group of male or female rats.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

No adverse test substance-related effects on mean food consumption were observed in any male or female group.

Food efficiency:

effects observed, treatment-related

Description (incidence and severity):

No test substance-related effects on food efficiency were observed in female rats in any dose group. In male rats, mean overall food efficiency (test days 0-91) was significantly decreased in animals dosed with 500 mg/kg bw/day compared with the control group. This significant decrease in food efficiency was still exhibited following a 30-day recovery period, but revealed within 90-day recovery period. This effect seems to be related to the body weight decrease in male rats at same dose level, since the food consumption was not effected by the test substance.

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

not examined

Ophthalmological findings:

no effects observed

Description (incidence and severity):

No test substance-related ophthalmology findings were observed in any male or female group. The observations noted in 1/20 males and 1/20 females at 20 mg/kg bw/day (keratitis of the cornea, retinal degeneration, shrunken globe-phthisis bulbi) did not exhibit a dose response and no statistically significant increase in incidence was observed. The effect were regarded as incidental.

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

Toxicologically relavant hematologic effects were observed at 500 mg/kg bw/day, manifested by mild to moderate decreases (75%, 73%, and 78% of control, respectively, for males; 92%, 94%, and 96% of control, respectively, for females) in red blood cell mass parameters (red blood cell count, hemoglobin and hematocrit) on test days 43/44 and futher decreased (69%, 64%, and 69% of control, respectively, for males; 82%, 85%, and 87% of control, respectively, for females) on day 92/93 (males/females). An erythroid regenerative response was observed at 500 mg/kg bw/day by increased reticulocyte count on test days 43/44 and 92/93 (males/females). Red blood mass parameters were essentially comparable to values for concurrent controls following 30-day and 90-day treatment-free recovery periods, indicating the reversibility of the treatment-related hematology changes at 500 mg/kg bw/day.
(see Table 1 and 2 in "any other information on results")

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

There were no changes in clinical chemistry parameters indicative of treatment-related organ toxicity. Aspartate aminotransferase (AST) was minimally increased at test day 43 in males administered 500 mg/kg bw/day (133% of control) and at test day 92 in males administered 100 and 500 mg/kg bw/day (125% and 139% of control, respectively). Alanine aminotransferase (ALT) was minimally increased at test day 43 in males administered 20 and 500 mg/kg bw/day (125% and 138% of control, respectively). However, there was no dose-related pattern for the increase in ALT in treated males at test day 43. Changes in ALT and AST werereversible following the 30- and 90-day recovery period. There were no treatment-related adverse findings at doses of 100 mg/kg bw/day or below.
The increases in group mean AST and ALT in males at 100 and 500 mg/kg bw/day at test days 43 and 92 are considered treatment related but were not toxicologically relevant in view of the generally minimal nature of the differences from controls and also as the underlying pathophysiologic process likely relates to an adaptive response in the liver (i.e., enzyme induction).

Urinalysis findings:

no effects observed

Behaviour (functional findings):

no effects observed

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

Test substance-related effects on organ weights were present in the liver and kidney.
Liver: Statistically significant increased weights (relative to body weight) were present in main study males and females at 500 mg/kg bw/day (163% and 137% for males and females, respectively) and in males at 100 mg/kg bw/day (111%), when compared to the control group. Liver weight effects in the 500 mg/kg bw/day groups showed some, but not complete, recovery after 30- and 90-day treatment-free periods; these changes were reversible after 90 days recovery in the 100 mg/kg bw/day males. These liver effects were consistent with secondary responses to the xenobiotic induction of metabolizing enzymes and/or peroxisome proliferation and were not considered adverse. No test-substance related liver effects were observed in male or female rats at 20 mg/kg bw/day.
Kidney: Weights (relative to body weight) were statistically significant increased in 500 mg/kg bw/day main study males and females (117% and 1116% for males and females, respectively) and in 100 mg/kg bw/day males (113%), when compared to the control group. In 500 mg/kg bw/day group, kidney weight effects were reversible following the 30-day recovery period in females but not in males. In 100 mg/kg bw/day group effects were reversible following the 30-day recovery period in males. Kidney weights were not determined in the 90-day recovery groups. Kidney weight changes were not associated with microscopic or clinical pathology changes indicative of renal toxicity. Therefore, these changes were considered to be treatment related, but non-adverse. No test-substance related kidney effects were observed in male or female rats at 20 mg/kg bw/day.

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

Pale discoloration of the liver was present in 3/10 male rats in the 500 mg/kg bw/day main group (sacrificed at the end of the dosing period). All other gross observations occurred in low incidences and were consistent with normal background lesions in rats of this age and stock.

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

Test substance-related microscopic findings were present in the nose, liver, and thyroid gland. In addition, changes occurring secondary to effects on red blood cells were observed in bone marrow and spleen.

Nose: Test substance-related adverse findings were present in the nose of male and female rats administered 100 or 500 mg/kg bw/day. Nasal lesions were characterized by degeneration and atrophy of olfactory epithelium, turbinate adhesions and respiratory metaplasia. Minimal changes, including turbinate adhesions, respiratory metaplasia, and microcysts within olfactory epithelium, were present in the nose following 30 and 90 days recovery. No test-substance related adverse findings were present in the nose of male or female rats at 20 mg/kg bw/day. (see Table 3 in "any other information on results")
Nasal lesions are considered to be a local effect based on irritant/corrosive properties of the test substance. Respiratory tissues might incidentally be exposed to the test substance following gavage procedure and/or by reflux of test substance after gavage.

Liver: Microscopic hepatocellular hypertrophy was present in males and females at 500 mg/kg bw/day and in males at 100 mg/kg bw/day. These liver effects were consistent with secondary responses to the xenobiotic induction of metabolizing enzymes and/or peroxisome proliferation and were considered to be treatment related but not toxicologically relevant.

Thyroid gland: Potentially adverse findings of follicular cell hypertrophy were present at the 500 mg/kg bw/day dose in the thyroid gland of male and female rats at the end of dosing and after 30-days recovery. After 90-days recovery, changes were limited to equivocal hypertrophy in 2/10 males in the 500 mg/kg bw/day group. No test-substance related effects on follicular cell hypertrophy were observed in male or female rats at 100 mg/kg bw/day and at 20 mg/kg bw/day.
Thyroid hypertrophy is a common finding in rats in association with induction of hepatic microsomal enzymes, and in this study was seen only at doses that also produce liver hypertrophy. Therefore, thyroid follicular cell hypertrophy in the current study was considered potentially adverse in the species tested.

Hematopoetic tissues: Increased incidences of minimal to mild splenic extramedullary hematopoiesis and /or erythroid hyperplasia in bone marrow were present in 500 mg/kg bw/day males and females sacrificed at the end of the dosing period. These changes were correlative to changes in red cell mass parameters (see Clinical Biochemistry findings) and are an expected finding as part of a regenerative red cell response. As such, these findings were not considered to be primary adverse effects. Consistent with the hematological findings, no compound-related changes were present in spleen or bone marrow in the male or female 500 mg/kg bw/day groups following 30-days recovery. Therefore, these tissues were not evaluated in the 90-day recovery groups.

Histopathological findings: neoplastic:

no effects observed

Key result

Dose descriptor:

NOAEL

Remarks:

local

Effect level:

20 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No adverse effects observed at 20 mg/kg bw/day.

Dose descriptor:

LOAEL

Remarks:

local

Effect level:

100 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

histopathology: non-neoplastic

Key result

Dose descriptor:

NOAEL

Remarks:

systemic

Effect level:

100 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No adverse systemic effcts observed at 100 mg/kg bw/day.

Dose descriptor:

LOAEL

Remarks:

systemic

Effect level:

500 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

body weight and weight gain

food efficiency

haematology

histopathology: non-neoplastic

Key result

Critical effects observed:

yes

Lowest effective dose / conc.:

500 mg/kg bw/day (actual dose received)

System:

haematopoietic

Organ:

bone marrow

spleen

Treatment related:

yes

Dose response relationship:

yes

Relevant for humans:

not specified

Table 1. Effects on haematological values in male rats (upper rows: mean value; lower rows: SD(n))

)Dose (mg/kg bw/day)	0	20	100	500
Red blood cell count (x10E6/µL)
Day 43	8.39	8.52	8.22	6.32*
	0.40(10)	0.27(10)	0.40(10)	1.22(10)
Day 92	8.89	8.95	8.46	6.09*
	0.36(10)	0.34(10)	0.41(10)	1.27(10)
Day 120	8.60	-	-	8.87@
	0.48(10)			0.26(10)
Day 183	8.57	8.51	8.35	8.11
	0.29(10)	0.49(10)	0.34(10)	1.07(10)
Hemoglobin (g/dL)
Day 43	15.3	15.6	14.7	11.2@
	0.7(10)	0.4(10)	0.7(10)	2.7(10)
Day 92	15.4	15.5	14.5	9.9@
	0.5(10)	0.4(10)	0.7(10)	2.8(10)
Day 120	15.5	-	-	15.7
	1.0(10)	-	-	0.4 (10)
Day 183	15.2	15.1	14.8	14.6
	0.3(10)	0.7(10)	0.5(10)	1.4(10)
Hematocrit (%)
Day 43	48.2	48.9	46.4	37.8*
	2.5(10)	1.5(10)	2.4(10)	7.1(10)
Day 92	49.0	49.2	46.2	34.0@
	1.4(10)	1.7(10)	1.8(10)	7.8(10)
Day 120	45.7	-	-	47.7*
	2.2(10)	-	-	1.4(10)
Day 183	45.7	45.9	45.1	44.6
	1.2(10)	2.6(10)	2.0(10)	3.4(10)
Absolute reticulocyte count (x10E3/µL)
Day 43	186.1	184.5	197.9	515.8@
	27.1(10)	30.6(10)	37.2(10)	201.1(10)
Day 92	174.0	149.7	167.3	539.8@
	30.4(10)	24.0(10)	23.6(10)	212.0(10)
Day 120	202.3	-	-	118.4*
	41.3(10)			23.7(10)
Day 183	171.5	165.9	169.8	233.4
	21.6(10)	22.2(10)	25.9(10)	182.2(10)

* Statistically significant difference from control at p < 0.05 by Dunnett/Tamhane-Dunnett test.

@ Statistically significant difference from control at p < 0.05 by Dunn's test.

Table 2. Effects on haematological values in female rats (upper rows: mean value; lower rows: SD(n))

Dose (mg/kg bw/day)	0	20	100	500
Red blood cell count (x106/µL)
Day 44	8.13	8.27	8.10	7.38*
	0.31(10)	0.31(10)	0.40(10)	0.47(9)
Day 93	8.34	8.53	8.32	6.85*
	0.43(10)	0.52(10)	0.27(10)	0.63(9)
Day 121	8.25	-	-	8.28
	0.43(10)	-	-	0.42(10)
Day 184	8.02	7.67	7.85	7.85
	0.24(10)	0.36(10)	0.33(9)	0.35(10)
Hemoglobin (g/dL)
Day 44	15.2	15.5	15.2	14.3*
	0.5(10)	0.4(10)	0.5(10)	0.6(9)
Day 93	15.6	15.8	15.6	13.3*
	0.7(10)	0.8(10)	0.4(10)	0.9(9)
Day 121	15.7	-	-	16.2
	0.7(10)	-	-	0.5(10)
Day 184	15.0	14.8	14.7	15.0
	0.5(10)	0.6(10)	0.6(9)	0.8(10)
Hematocrit (%)
Day 44	46.2	46.8	46.3	44.5
	1.7(10)	1.2(10)	1.9(10)	2.1(9)
Day 93	47.2	47.7	47.1	41.2*
	2.3(10)	2.5(10)	1.4(10)	2.4(9)
Day 121	45.5	-	-	46.4
	2.2(10)			1.5(10)
Day 184	45.6	44.3	44.4	45.1
	1.6(10)	2.6(10)	1.4(9)	2.0(10)
Absolute reticulocyte count (x10E3/µL)
Day 44	183.1	184.8	184.9	319.0*
	40.5(10)	28.9(10)	39.4(10)	70.3(9)
Day 93	142.9	153.4	161.8	401.7@
	34.3(10)	36.8(10)	43.5(10)	158.9(9)
Day 121	158.8	-	-	119.9*
	43.3(10)	-	-	33.3(10)
Day 184	149.5	151.5	152.7	138.3
	26.7(10)	26.4(10)	38.2(9)	35.0(10)

* Statistically significant difference from control at p < 0.05 by Dunnett/Tamhane-Dunnett test.

@ Statistically significant difference from control at p < 0.05 by Dunn's test.

Table 3. Incidences of test substance-related nasal lesions in male and female rats

	Male				Female
Dose (mg/kg bw/day)	0	20	100	500	0	20	100	500
Number examined	10	10	10	10	10	10	10a	10
Degeneration/Atrophy, OE
Main study	0	0	4	7	0	0	5	4
30-day recovery	0	-	-	0	0	-	-	0
90-day recovery	0	0	0	0	0	0	0	0
Adhesions, turbinates
Main study	0	0	0	3	0	0	0	3
30-day recovery	0	-	-	2	0	-	-	3
90-day recovery	0	0	0	5	0	0	0	2
Respiratory Metaplasia
Main study	0	0	0	4	0	0	1	7
30-day recovery	0	-	-	8	0	-	-	8
90-day recovery	0	0	0	8	0	0	0	4
Intraepithial microcysts, OE
Main study	0	0	0	0	0	0	0	0
30-day recovery	0	-	-	3	0	-	-	1
90-day recovery	0	0	0	4	0	0	0	0

a    In 100 mg/kg bw/day females, the number examined was 11 and 9 for the main study and 90-day recovery groups, respectively.

OE = olfactory epithelium

Underlined values were interpreted to be test substance-related effects by the study director.

- not evaluated

Conclusions:

Under the conditions of this study, the local NOAEL for subchronic toxicity was 20 mg/kg bw/day, based on microscopic pathology of the nasal tissue (olfactory epithelium degeneration and atrophy) observed at 100 and 500 mg/kg bw/day. Considering the irritating properties of the test substance, local effects observed are assumed to result from gavage procedure and/or reflux of the test substance after gavage. The systemic NOAEL was 100 mg/kg bw/day based on decreased red blood cell parameters, splenic extramedullary hematopoiesis and erythroid hyperplasia in bone marrow and follicular cell hypertrophy in the thyroid gland observed at 500 mg/kg bw/day.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Report date:

2009

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 415 [One-Generation Reproduction Toxicity Study (before 9 October 2017)]

Version / remarks:

adopted in 1983

Deviations:

no

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Sodium undecafluorohexanoate

EC Number:

220-881-7

EC Name:

Sodium undecafluorohexanoate

Cas Number:

2923-26-4

Molecular formula:

C6HF11O2.Na

IUPAC Name:

sodium undecafluorohexanoate

Test animals

Species:

rat

Strain:

other: Crl:CD(SD)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Inc., Raleigh, North Carolina, U.S.A.
- Females nulliparous and non-pregnant: not specified
- Age at study initiation: 6 - 8 weeks
- Housing: Acclimation period and pre-mating: 1 animal per cage; Mating: 1:1; Lactation: neonates were kept with the dam in polycarbonate cages; post-weaning period/non-mating period: amimals were kept individually in stainless wire mesh cages and in polycarbonate cages
- Diet: PMI® Nutrition International, LLC Certified Rodent LabDiet® 5002, ad libitum
- Water: tap water, ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 - 26
- Humidity (%): 30 - 70
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
Dose concetrations in vehicle: 4, 20 and 100 mg/mL
Dose volume: 5.0 mL/kg bw

Details on mating procedure:

- M/F ratio per cage: 1:1
- Length of cohabitation: 2 weeks
- Proof of pregnancy: vaginal plug or sperm in vaginal smear referred to as day 0 of pregnancy
- After successful mating each pregnant female was caged individually

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Concentration verification and stability of dosing formulations was measured from 4 different collecting time points during the study: near the beginning, near the middle (approximately test day 45) and near the end of the exposure phase of the study and after completion of dosing to lactating females as described in the previous section (approximately test day 117). Concurrent with dosing formulation analyses, recovery of the test substance from spiked NANOpure® water was tested at the low level (approximately 4 mg/mL), the middle level (approximately 20 mg/mL), and the high level (approximately 100 mg/mL) to confirm the analytical method.
Data from the analysis of the samples during the study indicate that the test substance was at the targeted concentrations, mixed uniformly, and stable under the conditions of the study. Test substance was not found in the control group samples.

Duration of treatment / exposure:

P0 females: approx. 126 days
P0 males: approx. 110 days

Frequency of treatment:

once daily, 7 days/weeks

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

20 mg/kg bw/day (actual dose received)

Doses / concentrations 2

Dose / conc.:

100 mg/kg bw/day (actual dose received)

Doses / concentrations 3

Dose / conc.:

500 mg/kg bw/day (actual dose received)

No. of animals per sex per dose:

20

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Dose level selection was based on the results of a preliminary dose range-finding study, were animals were orally exposed to 0, 500 and 1000 mg/kg bw/day for 2 weeks. Test substance-related effects on hematology (decreased red cell mass, hemoglobin and hematocrit; increased reticulocytes and platelets) and clinical chemistry (increased AST and ALT) occurred in male rats dosed with 500 or 1000 mg/kg bw/day. In addition, an increase in absolut and relative liver weights was observed in animals receiving 500 or 1000 mg/kg bw/day test substance. Animals dosed with 500 and 1000 mg/kg bw/day test substance exhibited mildly decreased cholesterol and triglyceride levels and decreases in total protein due to decreases in globulin. Therefore, 500 mg/kg bw/day was selected as the highest dose level for the main study.

Examinations

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily for mortality/moribundity and general condition; additionally 2 h post dosing ± 30 min for acute clinical signs of toxicity
- Cage side observations included: general condition and clinical signs, mortality/viability

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: weekly
- Observations included: evaluation of fur, skin, eyes, mucous membranes, occurrence of secretions and excretions, autonomic nervous system activity (lacrimation, piloerection, and unusual respiratory pattern), changes in gait, posture, response to handling, presence of clonic, tonic, stereotypical, or bizarre behavior

BODY WEIGHT: Yes
- Time schedule for examinations: once/ week; Female rats were weighed weekly during pre-mating and mating and on gestation days 0, 7, 14, and 21 and lactation days 0, 7, 14, and 21.

FOOD CONSUMPTION: Yes
Females:
Premating dosing period: weekly, ending approx. test day 70; cohabitation period: food consumption was not determined; gestation period: on gestation days 0, 7, 14, and 21; Lactation period: food consumption was not determined
Males: Premating dosing period: weekly, ending approx. test day 70; cohabitation period: food consumption was not determined

FOOD EFFICIENCY: Yes
- Mean daily food efficiency (g body weight gain/g food consumed) was calculated from food consumption and body weight data.

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No

Oestrous cyclicity (parental animals):

Vaginal smears were collected daily from all parental female rats in order to determine the stages of the estrous cycle. Vaginal smears were collected beginning 3 weeks prior to mating, and continuing until copulation was confirmed, or the cohabitation period has ended. Vaginal smears were also collected from all parental female rats at the time of sacrifice to determine the stage of estrous cycle. Vaginal lavage samples were examined microscopically for determination of the stage of the estrous cycle (diestrus, estrus, proestrus).

Sperm parameters (parental animals):

Parameters examined in P0 male parental generations:
Sperm motility, morphology, and concentration in the cauda epididymis, and spermatid concentration in the testis for first 10 surviving parental rats in each group.

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 offspring:
number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, weight gain, food consumption, physical or behavioural abnormalities, vaginal patency, preputial separation

GROSS EXAMINATION OF DEAD PUPS: yes, for external and internal abnormalities

Postmortem examinations (parental animals):

SACRIFICE
All parental rats were examined grossly, including females that died, were sacrificed in extremis, or were accidentally killed prior to the end of lactation, and those for which mating did not result in production of offspring.

GROSS NECROPSY
Gross necropsy consisted of external and internal examinations including the cervical, thoracic, and abdominal viscera; The uteri of all cohabited females were examined for the presence and number of implantation sites.

HISTOPATHOLOGY / ORGAN WEIGHTS
The following organs were collected and weighed: liver, brain, thyroid gland, ovaries (with oviducts), uterus (with cervix), testes, prostate, seminal vesicles and epididymides
Reproductive organs of animals with impaired reproductive performance (e.g., failure to conceive or deliver healthy offspring, or effects on estrous cyclicity or sperm parameters) were evaluated microscopically.

Postmortem examinations (offspring):

SACRIFICE
Three F1 weanlings/sex/litter (randomly selected), litter size permitting, were sacrificed by carbon dioxide asphyxiation on postnatal day 21. All remaining F1 generation adult rats were sacrificed by carbon dioxide asphyxiation and exsanguination on postnatal day 60.

GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations including the cervical, thoracic, and abdominal viscera.

HISTOPATHOLOGY / ORGAN WEIGTHS
The following tissues from F1 adults were weighed: testes, epididymides, seminal vesicles (with coagulating glands and their fluids), prostate, uterus (with cervix), thyroid gland, liver, brain, and gross lesions. No microscopic examination was conducted on tissues from F1 adults.

Statistics:

Male and female parental data were evaluated separately. For litter parameters, the proportion of affected pups per litter or the litter mean were used as the experimental unit for statistical evaluation. For each parameter analyzed with a trend test, the test was applied to the data sequentially. If a significant dose-response was detected, data from the top dose group was excluded and the test repeated until no significant trend was detected. The level of significance selected was p < 0.05.
For further information see "Any other information on material and methods" Table 1.

Reproductive indices:

Mating index (%) = (number of females with confirmed mating / number cohabited) x 100
Female fertility index (%) = (number of pregnant females* / number of females with confirmed mating) x 100
Gestation index (%) = (number of litters with at least 1 live pup / number of litters) x 100
Implantation efficiency (%)** = (number of pups born / number of implantation sites) x 100

*Including delivery of a litter or uterine implantation sites.
**To be determined for each litter. Mean and standard deviation for each dose level was calculated.

Offspring viability indices:

Pups born alive (%)** = (number of pups born alive on postnatal day 0 / total number of pups born) x 100
Viability index (%)**# = (number of pups surviving on postnatal day 4 preculling / number of pups born alive on postnatal day 0) x 100
Lactation index (%)**# = number of pups alive at weaning (day 21 postpartum) / number of pups alive day 4 postculling x 100
Litter survival (%)# = number of litters weaned / number of viable litters delivered x 100

**To be determined for each litter. Mean and standard deviation for each dose level was calculated.
# Excluding litters sacrificed due to death of dam during lactation.

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

Clinical signs of toxicity were observed in parental males and females at 500 mg/kg bw/day and consisted of increased incidence of stained skin/fur in 13/20 (vs. 0 in control) males and 3/20 (vs. 0 in control) females (during daily cage site observations only).

Dermal irritation (if dermal study):

not examined

Mortality:

mortality observed, non-treatment-related

Description (incidence):

There was no test substance-related mortality in parental females at any level tested. Mortality that occurred within the first 2 weeks of study was not considered test substance related since the exposure duration was minimal and there were no specific causes of death determined at pathology. One male rat of 20 mg/kg bw/day group was found dead on Day 23. Gross pathology examinations revealed kidney, urinary bladder, and prostate abnormalities for this male. The mortality observed at 20 mg/kg bw/day was not considered treatment related since it occurred at a low incidence and there was no other evidence of toxicological significance at this treatment level.
Mortality occurred in 5 females at 500 mg/kg bw/day. Two females died during the first week (Day 4 and 9) of test substance administration and the other died during lactation (day 114 / lactation day 18). Prior to death, one animals was hunched over and one animal was dehydrated and had stained fur. One animals (lactation day 18) exhibited signs of dehydration prior to death. This death was not considered toxicologically relevant since it occurred for only one animal after approximately 16 weeks of treatment and there were no specific causes of death observed at pathology. In addition, 2 deaths on Day 52 and 84 were attributed to mechanical trauma (paw and esophagus, respectively).

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Males:
Test substance-related effects on body weight parameters were observed in males at 500 mg/kg bw/day. At 500 mg/kg bw/day, body weight was 7-15% lower than control on test days 49-105; contributing to an overall body weight gain reduction (29%) as compared to the control group. At 100 mg/kg bw/day, body weights were 7% lower than the control group on test days 91-105. Overall body weight gain (TD 0 to 105) was reduced 12%; compared to control. The body weight decrease was considered toxicologically relevant in 500 mg/kg bw/day group only, since the difference was significant and >10% compared with control.

Females:
There were no test substance-related effects on body weights or weight gains in females during the premating period at any level tested. Instances of statistically significant increases in body weight or body weight gain that occurred at an intermediate level, but not the high level, were considered unrelated to the test substance and therefore not toxicologically significant.
During the gestation period, there were was a test substance-related reduction in mean maternal body weight gain at 500 mg/kg bw/day. This reduction was observed during the first week of gestation when mean body weight gain was 29% lower than the control group. Maternal body weight was unaffected throughout gestation for all treatment levels. This reduction in weight gain was considered not toxicologically relevant, since it was transient over first week of gestation only.
During the lactation period, there were test substance-related effects on body weight gains at 100 and 500 mg/kg bw/day. In both levels, a lack of expected reduced body weight gain was observed throughout lactation. Animals at 100 and 500 mg/kg bw/day gained an average of 20.4 and 24.9 g (respectively) throughout lactation compared with a gain of 5.1 g in the control group. Instances of statistically significant increases in body weight that occurred at 100 mg/kg bw/day, but not at 500 mg/kg bw/day, were considered unrelated to the test substance and therefore not toxicologically significant. There were no statistically significant, test substance- related effects on body weight at any dose level tested throughout lactation.

(refer to table 2)

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

Males: There were no test substance-related effects on food consumption in male rats at any dose level tested.

Females: Food consumption was reduced (11%) compared with the control group at 500 mg/kg bw/day during the first week of gestation. There were no statistically significant, test substance-related effects on food consumption parameters during premating at any level tested.

Instances of statistically significant increases in food consumption parameters that occurred at 100 mg/kg bw/day, but not at 500 mg/kg bw/day, were considered unrelated to the test substance and therefore not toxicologically significant.

Food efficiency:

effects observed, treatment-related

Description (incidence and severity):

Males: Overall food efficiency was reduced (24% lower than control) at 500 mg/kg bw/day compared with the control as a result of reduced body weight gain throughout the premating period. Instances of statistically significant decreases in food consumption that occurred in the 100 mg/kg bw/day group, but not in the 500 mg/kg bw/day group, were considered unrelated to the test substance and therefore not toxicologically significant.

Females: Food efficiency was reduced (19%) compared to the control group at 500 mg/kg bw/day during the first week of gestation as a result of reduced body weight gain for this interval.

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:

not examined

Histopathological findings: non-neoplastic:

effects observed, non-treatment-related

Description (incidence and severity):

4/20, 4/20, 1/20 and 0/20 females in the 0, 20, 100, and 500 mg/kg bw/day groups, respectively, were not pregnant. Three females (each one of control group, 20 mg/kg bw/day and 100 mg/kg bw/day group, respectively) had ovaries with an absence of corpora lutea. Therefore, these females did not ovulate. A control male had severe diffuse testicular atrophy and severe oligospermia in the epididymus. For the remaining animals, there were no microscopic findings correlative to their reproductive failure.

Histopathological findings: neoplastic:

no effects observed

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

no effects observed

Reproductive function: sperm measures:

no effects observed

Reproductive performance:

effects observed, non-treatment-related

Description (incidence and severity):

There was a statistically significant increase in the mean number of implantation sites observed in females administred 100 mg/kg bw/day. However, no dose relationship was evident, and this effect was therefore not considered test substance related.
Litter size, sex ratio and pup survival: There were no test substance-related effects on litter size, sex ratio, or pup survival at birth at any level tested. Statistical significant increases in litter size that occurred at 100 mg/kg bw/day, but not at 500 mg/kg bw/day, were considered incidental and unrelated to the test substance treatment.

Effect levels (P0)

1

Key result

Dose descriptor:

NOAEL

Remarks:

fertility

Effect level:

500 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No adverse effects on parental fertility observed at highest dose tested.

2

Key result

Dose descriptor:

NOAEL

Remarks:

systemic

Effect level:

100 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male

Basis for effect level:

other: No adverse effects observed at 100 mg/kg bw/day.

3

Dose descriptor:

LOAEL

Remarks:

systemic

Effect level:

500 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male

Basis for effect level:

body weight and weight gain

4

Key result

Dose descriptor:

NOAEL

Remarks:

systemic

Effect level:

500 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

female

Basis for effect level:

other: No adverse effects observed up to the highest dose tested..

Target system / organ toxicity (P0)

Key result

Critical effects observed:

no

Results: F1 generation

General toxicity (F1)

Clinical signs:

no effects observed

Description (incidence and severity):

There were no test substance-related clinical signs of toxicity observed in F1 males or females at any level tested throughout postweaning.

Dermal irritation (if dermal study):

not examined

Mortality / viability:

mortality observed, non-treatment-related

Description (incidence and severity):

In control group, mortality occurred for 1/20 male (FD 17) and 1/20 female (TD1). Prior to death, one animal had a swollen head and abnormal gait and both animals in the control group revealed cranial fluid and soft brain at pathology.
In 20 mg/kg bw/day group, mortality was limited to 1/20 female (TD 18). There were no adverse clinical signs or pathology findings observed for this animal.
In 500 mg/kg bw/day group, 3/20 males (TD 2, 6 and 7) and 1/20 female (TD 6) died. 2 animals revealed the same pathology findings as the control animals along with dilated urinary bladder and heart cysts and apex. Both of these animals originated from the same litter. The other 2 deaths at 500 mg/kg bw/day also originated from the same litter. Prior to death, one animal had a missing eye and one was immobile, cold to touch, and had enopthalmus. Pathology findings included a small eye and optic nerve for one animal.
In conclusion, there were no test substance-related mortality observed in F1 males or females at any level tested throughout postweaning. The cases of mortality were incidental and not considered treatment related.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Lactation period:
Pup weights during lactation period: Statistically significant reductions in pup weights were observed (17-18% lower than control group) at 500 mg/kg/day throughout lactation (PND 0-21).

Post-weaning period:
Males: Body weights were significantly reduced compared with the control group throughout the post-weaning period at 500 mg/kg bw/day.This was due to a lower body weight of the high-dose group on Day 0 post-weaning; the body weight gain of the high-dose pups was comparable to the control group. During the first 4 weeks post-weaning, the body weights were 7-16% lower than control group. Body weight gain during the first week after weaning was 16% lower than control group at 500 mg/kg bw/day and the difference was reduced over the following three weeks. Body weight of this group was similar to the control group from Day 28 after weaning. Overall body weight gain (post-weaning days 0-39) was comparable across dose levels.
Females: Body weights were reduced (11%-12% lower than control group) during the first week (post weaning) at 500 mg/kg bw/day. Body weight of this group was similar to the control group from Day 14 after weaning. There were no test substance-related effects on body weight gains in F1 adult female rats at any level tested.
In conclusion, the differences in body weight and body weight gain observed between the high-dose group and control group are not considered to be toxicologically relevant, since the effects were transient. During the postweaning period, the high-dose offspring increased the body weight gain until the body weights were comparable with the control.

(refer to table 2)

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

Males: At 500 mg/kg bw/day, food consumption was statistically significantly reduced (13% lower than control group) during the first week after weaning.

Females: There were no test substance-related effects on food consumption parameters at any level tested in F1 adult female rats.

Food efficiency:

no effects observed

Description (incidence and severity):

There were no effects on food efficiency at any level tested in F1 adult male and female rats.

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

Sexual maturation:

no effects observed

Organ weight findings including organ / body weight ratios:

effects observed, non-treatment-related

Description (incidence and severity):

There were no test substance-related organ weight changes in F1 adult males or females. Testes weight and relative testis weight (relative to bw) were decreased by 7 and11% (statistically significant) in the 20 and 500 mg/kg bw/day males, respectively, compared with the control. However, this effect was not dose related, as there were no statistically significant changes in testes weight in the 100 mg/kg bw/day group. In addition, these effects in 100 and 500 mg/kg bw/day group are not considered toxicologically relevant, since they are < 20% different from control.
Statistically significant decreases in epididymides weights relative to body weight (in 20 mg/kg bw/day males) and relative to brain weight (in 100 mg/kg bw/day males) were not dose related and were thus not considered treatment related.

Gross pathological findings:

no effects observed

Description (incidence and severity):

There were no treatment-related gross findings. All gross observations in the respective generations and life stages occurred in low incidences and were consistent with normal background lesions in rats of this strain.

Histopathological findings:

not examined

Developmental neurotoxicity (F1)

Behaviour (functional findings):

not examined

Developmental immunotoxicity (F1)

Developmental immunotoxicity:

not examined

Effect levels (F1)

Key result

Dose descriptor:

NOAEL

Remarks:

development

Generation:

F1

Effect level:

500 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No adverse effects on development observed at 500 mg/kg bw/day.

Target system / organ toxicity (F1)

Key result

Critical effects observed:

no

Overall reproductive toxicity

Key result

Reproductive effects observed:

no

Any other information on results incl. tables

 Table 2. Mean (±SD) body weight gains (g) in P1 and F1 rats and body weights (g) of F1 pups.

Group	Day		Dosage (mg/kg bw/day)
			Control	20	100	500
P1 males	Test	0 - 105	340 ± 44	329 ± 47	299 ± 43*	241 ± 40*
P1 females	Gestation	0 - 7	36 ± 10	38 ± 8	37 ± 10	25 ± 8*
		0 - 21	140 ± 25	145 ± 16	147 ± 23	134 ± 19
	Lactation	0 - 21	5.1 ± 26	7.4 ± 20	20 ± 15	25 ± 12*
F1 pups	Postnatal	0	7.1 ± 0.9	6.8 ± 0.6	6.3 ± 0.4	5.8 ± 0.4^#
		7	18 ± 2.7	18 ± 2.2	17 ± 1.3	15 ± 1.4^#
		14	36 ± 3.4	37 ± 3.0	34 ± 2.6	30.0 ± 2.5^#
		21	59.6 ± 5.3	62 ± 5.0	57 ± 5.3	49 ± 4.1^#
F1 males	Postweaning^a	0 - 39	320 ± 25	327 ± 42	320 ± 27	321 ± 25
F1 females	Postweaning^a	0 - 39	183 ± 21	178 ± 18	173 ± 21	183 ± 24

^aAge of animals at postweaning day 0 = 21 days old.

* Statistically significant difference from control at p < 0.05 by Dunnett/Tamhane–Dunnett.

# Statistically significant difference from control at p < 0.05 by analysis of covariance and Dunnett–Hsu.

For detailed result tables of mean body weight and body weight gain of offspring during lactation and after weaning see 'attached back ground material'.

Applicant's summary and conclusion

Conclusions:

Under the conditions of this study, the NOAEL for systemic toxicity was considered to be 500 mg/kg bw/day for P0 females and 100 mg/kg bw/day for P0 males based on reduced body weight and body weight weight gain in males at 500 mg/kg bw/day. No adverse effects on parental fertility occurred up to the highest dose tested, therefore the NOAEL for fertility was 500 mg/kg bw/day. A transient reduction in body weight and body weight gain was observed in male and female F1 rats at 500 mg/kg bw/day during lactation period and first weeks post-weaning. These effects were considered not to be toxicologically relevant since they were transient. Further, no effects on other developmental parameters or teratogenicity were observed. Therefore, the NOAEL for developmental toxicity was considered to be 500 mg/kg bw/day. Sodium perfluorohexanoate is therefore concluded not to present a reproductive or developmental hazard and no classification is justified.