Table F.4.

Perfluoroalkane sulfonic acids (PFSAs): reproductive and developmental toxicity

Substance/ (Purity)	Species/Experimental design and doses	Observed effects	Highest dose with no effect (mg/kg bw per day)		Lowest dose with effect (mg/kg bw per day)		Serum/tissue levels of compound	Reference
			Mother	offspring	Mother	offspring
Perfluorobutane sulfonic acid (PFBS)
PFBS (97.9% pure, K+‐salt)	Sprague Dawley rats 2‐generation reproduction study according to OECD guideline 416 Exposure: 0, 30, 100, 300, 1,000 mg/kg bw per day, gavage. Parental (F0) animals (males and females, n = 29–30 per sex per group) dosed from 70 days prior to mating, females were continued through gestation and lactation. F1 offspring (n = 29‐30) was dosed from weaning (lactational day 22) onwards. F2 generation was exposed through placenta and lactation. Experiment was terminated at lactational day 22 of F2 generation animals	Increased liver weight Increased hepatocellular hypertrophy (P and F1 adult males) Increased incidence of mild microscopic findings in kidney No reproductive and developmental toxicity findings	100 100 100	1,000	300 300 300		Not determined	Lieder et al. (2009b)
PFBS (97% pure, K+‐salt)	ICR mice Dosing: 0, 50, 200 and 500 mg/kg bw per day from GD 1–20, orally. Only female offspring follow‐up reported. 30 dams per dose group, randomly allocated to three experimental subgroups per dose: group 1: perinatal survival and growth, pubertal onset, and ovarian and uterine development (10 dams, 50 female offspring per dose), group 2: hypothalamic–pituitary–gonadal hormone and hypothalamic– pituitary–thyroid hormone levels (10 dams, 30 PND 1 female offspring, 10 female PND 30 offspring, and 10 PND 60 female offspring), group 3: levels of serum PFBS (10 dams)	Decreased body weight from PND 1 to adulthood Delayed eye opening Delayed vaginal opening Impaired ovarian and uterine development Delayed oestrus cyclicity and reduced E2/increased LH Decreased T3/T4, increased TSH Decreased T3/T4, increased TSH at GD 20	50	50 50 50 50 50 50	200	200 200 200 200 200 200	Mean (10 dams) serum concentration (ng/mL) on GD 20, 12 h after last dosing, at 0, 50, 200 and 500 mg/kg bw per day: 1.73, 74, 332, 721	Feng et al. (2017)
Perfluorohexane sulfonic acid (PFHxS)
PFHxS (99.98% pure, K+‐salt)	Sprague‐Dawley rats, OECD guideline 422‐based design Exposure: 0, 0.3, 1, 3, 10 mg/kg bw per day by oral gavage to F0 (n = 15 + 3; main study + serum sampling) starting 14 days prior to mating until PND 22 or presumed gestation day 25 for rats without litter	No reproductive or developmental effect No treatment‐related effects in dams	10	10			Elaborated serum and liver values in dams and fetuses. Serum levels at study day 14 and GD 21 in dams were similar. For GD 21 females in serum and liver: 0 mg/kg per day: < LOQ of 0.1 μg/mL and 0.1 μg/g. 0.3 mg/kg per day: 3.32 μg/mL and 0.79 μg/g 1 mg/kg per day: 10.65 μg/mL and 2.61 μg/g 3 mg/kg per day: 32.75 μg/mL and 7.80 μg/g 10 mg/kg per day: 59.80 μg/mL and 16.53 μg/g	Butenhoff et al. (2009)
PFHxS, K+‐salt, purity > 98%	Wistar rats. Oral gavage from GD 7 until PND 22 (except for day of delivery). Range finding study (8 time mated rats per group): 0, 25, 46 mg/kg bw per day. Main study (16–20 time mated rats per group): 0, 0.05, 5, 25 mg/kg bw per day.	Increased liver weight. ‐ Males ‐ Females Pronounced reduction of T4 levels, detectable at different time points. Dams: Offspring: Mildly decreased body weight. Male pups Female pups	0.05	5 0.05 0.05 5 0.05	5	25 5 5 25 5	Serum levels in dam at PND 22 in the range finding study. 139 and 174 μg/mL in 25 and 45 mg/kg bw per day groups, respectively	Ramhøj et al. (2018)
K+PFHxS (98.9% pure)	Crl:CD1 (IRC) mice Study design according to OECD guideline 422, modified). Exposure: gavage at 0, 0.3, 1, 3 mg/kg bw per day (n=30+12 per treatment; main study + toxicokinetic arm). Main experiment: For F0 males treatment from 14 days prior to cohabitation for to at least 42 days total (one day post‐last dosing). Treatment of F0 females started 14 days prior to cohabitation with continuation through mating, gestation and lactation. F0 dams were sacrificed on lactation day 22 (one day after last dosing). F1 offspring, first exposure in utero and via lactation. After weaning (PND 22), F1 direct dosing for 14 days at maternal dose. Toxicokinetic experiment: 12 animals per sex and dose. Subset 1 (5/sex/dose group) daily oral gavage for 14 days prior to sacrifice. Subset 2 (7/sex/dose group) dosing for 14 days prior to cohabitation. Serum and liver samples were collected at study day 14 for both sexes, at study day 28 for males and GD 18 (for females). On GD 18, pooled fetal blood and fetal liver samples were collected	Reduced litter size (without impact on born pup to implant ratio) F0 animals: Increased mean and relative liver weight. F1 animals: Increased relative liver weight ♀ and ♂ Increased thyroid weight ♂	0.3 1 1	0.3	1 3 3	1	Serum and liver were measured at study day 14, GD 18 (toxicokinetic study arm) and lactational day 22 (main experiment) in dams and in F1 at GD 18 (pooled serum fetuses), PND 4 (pooled liver litter) and PND 21 and 36 (males and females). Serum and liver in dams, and serum from pooled fetus on GD18: 0 mg/kg per day: < 0.001 μg/mL, < 0.005 μg/g and < 0.001 μg/mL. 0.3 mg/kg per day: 16.8 μg/mL, 5.3 μg/g, and 20.8 μg/mL 1 mg/kg per day: 51.5 μg/mL, 15.1 μg/g and 62.3 μg/mL 3 mg/kg per day: 111.3 μg/mL, 88.4 μg/g and 137.7 μg/mL.	Chang et al. (2018)

bw: body weight; FSH: follicle stimulating hormone; GD: gestation day; LH: luteinizing hormone; LOQ: limit of quantification; PCNA: proliferating cell nuclear antigen; PND: postnatal day; T3: triiodothyronine; T4: thyroxine; TSH: thyroid stimulating hormone.