Document nmQLXoEgyZxwbzpJ3GLg9nKm6

AR226-3385 Subchronic, Developmental, and Reproductive Toxic Fluoroalkylethyl Phosphate Surfactant J.C. Stadler, DA. Delker, G.T. Makovec, J.F. Hansen, S.M. Munley, and E. Mvlchreest DuPont Haskell Laboratory for Health and Environmental Sciences, Newark, Delaware, Abstract A fluoroalkylethyl phosphate surfactant was evaluated in rats in 90-day, one-generati reproduction, and developmental toxicity studies. In the 90-day study, the test substa as a suspension in isopropanol (IPA) at oral doses ofO, 10, 60 or 300 mg/kg/day (35% 60 mg/kg/day IPA. There were no compound effects on mortality, clinical signs or ne Adverse effects on body weights, food parameters, red cell mass, and hepatic enzyme the highest two doses, and increases in thyroid weight and cellular hypertrophy were high dose. Adverse microscopic changes in the liver (focal hepatocellular necrosis) oc dose levels in males and at only the high dose in females. These effects were not seen dosed alone. Other non-adverse physiological changes included increased liver and ki hepatocellular hypertrophy, and an increased rate ofperoxisome proliferation. The N female rats in this study was 10 mg/kg/day. In the reproductive study, where surfacta 0, 75, 500 and 3500 mg/kg/day (20% a.i. in water, no IPA), parental body weights, we food parameters were reduced at the high dose, with males more affected than female no effects on reproduction parameters at any dose level, except for F^ pup weights, w progressively reduced from birth to weaning (92% and 76% of control, respectively) dose level. In these rats dosed with the active in water, thyroid hypertrophy and liver not observed. The NOEL for reproductive parameters was 500 mg/kg/day. In the dev study, doses were 625, 1250 or 2500 mg/kg/day (20% a.i. in water). At 2500 mg/kg/d maternal weight, food consumption, and fetal weight occurred. At 1250 mg/kg/day, m weight reductions occurred. No effects were seen at 625 mg/kg/day. Overall, the mo target of toxicity was the liver, but the test substance was not considered a reproducti developmental toxin. Introduction The objective of these studies was to evaluate the potential subchr developmental, and reproductive toxicity of a fluorotelomer-based phosphate surfactant in male and female rats. The surfactant is ge sold as a dispersion in isopropanol (IPA) and is used in numerous coating applications. In the subchronic study, the telomer product administered as formulated product that contained approximately active ingredient in IPA. In the developmental and reproductive t studies, the telomer surfactant was suspended in water, with a 20% concentration of active ingredient. The test material was administ gavage, the route selected as the most efficient way to deliver an a dosage. Separate range-finding studies were conducted to select appropriate dosages for each study. Study Designs All studies: Crl:CD(SD)IGS BR rats 90-Day Subchronic Toxicity Dosages: 0, 10, 60 or 300 mg/kg/day (35% a.i. in IP 60 mg/kg/day IPA No. of animals: 10/sex/group - main study, 10/sex/group intermediate, low - 1-month recovery, 5/sex/group - 3-month recovery Dosing Regimen: Parameters: 90 days all groups, 1-month and 3-month U.S. EPA Health Effects Test Guideline O 870.3100(1998) Developmental Toxicity Dosages: 0, 625, 1250, or 2500 mg/kg/day (20% a.i water) No. of animals: 22 pregnant females/group Dosing regimen: Gestation Days 6-20 Parameters: U.S. EPA Health Effects Test Guidelines 870.3700 Prenatal Developmental Toxicit Study Designs (Continued) One-Generation Reproduction Dosages: 0, 75, 500, and 3500 mg/kg/day (20% a.i. water) No. of animals: 20/sex/group Dosing Regimen: 70 days prior to mating and throughout m males and females Parameters: During gestation and lactation - dams onl Special Design (Figure 1) Pretest Reproductive Evaluation? Figure 1 - Study Design PI Adult Dosing Period Subchronic Study Results In-Life Parameters No mortality, clinical signs oftoxicity, or behavioral abnormalities Significant lower body weight at 300 mg/kg/day - males only (Figure 2) Low food consumption and food efficiency at 300 mg/kg/day - males Figure 2 " Male Body Weights 35 42 49 56 Days on Test Subchronic Study Results Liver Effects Effects commonly observed in rats with many organofluoride materials, peroxisome proliferation, hepatomegaly, and increased liver weights wer observed in this study Increased hepatic B-oxidation activity at 60 and 300 mg/kg/day - and females (Figure 3) Elevated liver weights and hypertrophy at 60 and 300 mg/kg/day and at 300 mg/kg/day in females (Figure 3) Elevated liver enzymes at all doses - males; at 300 mg/kg/day - Figure 3 - Beta Oxidation -- Males Liver Effects Beta Oxidation -- Females g 10 10 Days 90 Days 1-Month Recovery 3-Month Rec Liver Wei^it Effects in Female Rats Day 93 1-Month Recovery 3-Month Rec Subchronic Study Results Clinical Pathology Red cell mass parameters were affected at 60 and 300 mg/kg/day -- male (Table 1). These parameters were not affected in the reproduction study. TABLE 1 PARAMETERS OF RED CELL MASS Dosage (mg/kg/day): 10 60 300 Red Blood Cells Day 38 Day 92 1-month Recovery 3-month Recovery 101% 99% ND 97% 99% 97% ND 94% 96% 94% 90% 97% Hemoglobin Day 38 Day 92 1-month Recovery 3-month Recovery 99% 95% ND 97% 97% 95% ND 94% 93% 91% 94% 92% Hematocrit Day 38 Day 92 1-month Recovery 3-month Recovery 0% 96% ND 98% 98% 95% ND 94% 94% 91% 93% 93% Values are percent of control. Those in bold italics are statistically significant. ND = No data Subchronic and Reproductive Study Histopathology In the subchronic study with active suspended in alcohol there were adve lesions (focal necrosis) that did not occur in the study with active ingredi suspended in water. Similarly, chronic progressive nephropathy was evi the kidneys of animals dosed with active in water (reproductive study), b the subchronic study. Other histopathological lesions were similar. Table 2 - Histopathology Effects Dosage (mg/kg/day) Subchronic Adult Males Subchronic Adult Females PI Adult Males Fl Ad (Male and 3.5 (active in IPA) 15 (active in water) Liver: focal necrosis None Thyroid: hypertrophy Thyroid: hype 20 (active in IPA) Liver: hypertrophy. focal necrosis Thyroid: hypertrophy None 100 (active in water or Liver: hypertrophy, in IPA) focal necrosis Thyroid: hypertrophy Kidney: tubular hypertrophy Thyroid: hypertrophy Liver: hypertrophy Thyroid: hypertrophy Kidney: chronic progressive nephropathy Thyroid: hyper 700 (active in water) Liver: hypertrophy, Thyroid: hypertrophy Kidney: chronic progressive nephropathy Thyroid: hype Developmental Toxicity Results Maternal Toxicity Reductions in body weight parameters at 1250 and 2500 mg/kg/day. (Table 3) Effects on food consumption at 2500 mg/kg/day. Fetal Toxicity Reduced body weight parameters at 2500 mg/kg/day No malformations and no-compound-related variations at any dose lev TABLE 3 DEVELOPMENTAL EFFECTS Maternal Effects 2500 mg/kg/day Mean body weight (^7%) Adjusted body weight (4-26%) Food consumption (4^8%) ____1250 mg/kg/day 625 mg/kg/d Mean body weight (^5%) No effects Adjusted body weight (4^22%) Fetnl Effects Mean body weight (4-4%) No effects No effects Reproductive Toxicity Results Reduced body weight parameters at 3500 mg/kg/day - Pi and F and females Reduced food consumption at 3500 mg/kg/day - Pi and Fi male females Reduced body weight parameters and food consumption at 500 mg/kg/day - Pi males No effect on Pi generation sperm or estrous cycle parameters No effect on Pi generation mating, fertility, gestation length or implantation No effect on Fi litter size, sex ratio or lactation pup survival ind Reduced Fi pup birth and lactation weights at 3500 mg/kg/day TABLE 4 Pi GENERATION LITTERS Dose (mg/kg/day) 0 N 18 75 500 19 19 3500 15 Mean Number of Pups/Litter Bom 12.9 Bom Alive 12.8 Day4Precullmg 12.7 Day 4 Postculling 7.8 Day 21 7.8 11.4 11.3 11.3 7.3 7.3 12.4 12.3 12.3 8.0 8.0 13.0 12.5 12.4 7.9 7.9 Mean Pup Weights DayO (grams)__________________ 6.9 7.1 6.8 6.3 Day4Precullmg 11.1 11.6 11.5 10.1 Day 4 Postculling 11.0 11.6 11.4 10.1 Day 7 17.4 17.9 18.1 15.0 Day 14 36.9 35.3 35.7 28.4 Day 21 58.8 58.4 57.2 44.6 a Percent litters delivered having at least one live pup. b Mean percent survival from Day 4 Postculling to Day 21. c Percent litters bom with at least one pup alive on Day 21. Values in bold italics are statistically significant. Summary Subchronic Study 9 Based on hepatocellular necrosis at all dose levels, no NOEL wa in male rats Based on elevated liver enzymes and thyroid gland hypertrophy observed at 300 mg/kg/day, a NOEL of 60 mg/kg/day was estab for females Developmental Toxicity Study Based on decreases in body weight parameters at 1250 mg/kg/d NOEL for maternal effects was 625 mg/kg/day Based on decreases in fetal body weights at 2500 mg/kg/day, th NOEL was 1250 mg/kg/day Not a selective developmental toxin One-Generation Reproductive Study Based on reductions in mean pup weights throughout the entire period for F^ litters at 3500 mg/kg/day, the NOEL for reproduct effects was 500 mg/kg/day. However, based on thyroid follicula hypertrophy in P^ and F^ adult males at >75 mg/kg/day, a NOEL not determined for this study. Conclusions The target organs oftoxicity in the subchronic study were the li thyroid and red blood cells The phosphate surfactant is not a selective developmental toxin The reproductive toxicity of this material was limited to in utero lactational offspring growth impairment Acknowledgments We wish to thank the following individuals for their technical assi Nancy E. Everds for clinical pathology evaluations Linda A. Malley for neuropathology evaluations John C. 0'Connor for Biochemical Evaluations Primary Technicians: Robert E. Walker, Jr., Kellie Mcllhatton, Gregory L. Poindexter Poster Preparation: Maryanne M. Wilford Subchronic, Developmental and Reproductive Toxkity of a Fluoroalkylethyl Phosphate J.C. Stadler, D.A. Delker, G.T. Makovec, J.F. Hansen, S.M. Munley, and E. Mylchreest The DuPont Company, Haskell Laboratory for Health and Environmental Sciences, Newark, Delaware, USA ISSSAi b s t r a c t A fliioroalkylcttiyf phosphate surfaciant was evaluated in rate in 90-day, onegeneration reproduction, and developmental toxicity studies. In the 90-day study, the lest substance was given as a suspension in isopropanol <IPA) at oral doses ofO. 10,60 or 300 nig/kg/day (35% a.i.) and 60 mg/kg/day IPA. Tlicre were no compound effects on mortality, clinical signs or neurobehavior. Adverse effects on body weights, food parameters. red cell mass, and hepatic enzymes were seen at the highest two doses, Mid increases in thyroid weight Mid cellular hypertrophy were observed at the high dose. Adverse microscopic changes in the liver (focal hepalocelluiar necrosis) occurred al all dose levels in males and at only the high dose in females- These effects were not seen with IPA dosed alone. Other nonadverse physiological ctianges included increased liver and kidney weights, hepatocellular hypertrophy, and an increased rate of peroxisome proliferation. The NOEL for female rats in this study was 10 ing/kg/day. In the reproductive study, where surfactant doses were 0.75.300 and 3500 ing/kg/day (20% a.i- in water, no IPA), parental body weights, weight gains and food parameters were reduced al the high dose, with males more affected than females. There were no effects on reproduciion parameters at any dose level, except for F( pup weights, which were progressively reduced from birth 10 weaning (92% and 76% of control, respectively) at the high dose level. In these rats dosed with Hie active in walcr, thyroid hypertrophy and liver necrosis were not observed. The NOEL for reproductive parameters was 500 mg/kg/day. In the developmental study, doses were 625.1250 or 2500 mg/kg/day (20% a.i. in water). At 2500 mg/hg/day, effects on maternal weight, food consumption, and fetal weight occurred. At 1250 mg/kg/day, maternal body weight reductions occurred. No effects were seen al 625 mg/kg/day- Overall, Hie most sensitive target of toxicity was the liver, but the test substance was not considered a reproductive or developmental toxin, fUSSIntroduction The objective of these studies was to evaluate the potential subchronic, developmental, and reproductive toxicity of a fluorotelomer-bascd phosphate surfaciani in male and female rats. The surfactant is generally sold as a dispersion in isopropanol (IPA) and is used in numerous surface coating applications. In the subchronic study, the Iclomer product was administered as formulaicd product lliat contained appro?, imaiety 35% active ingredient in IPA. In the developmental and reproductive toxicity sludies, the telomer surfaciant was suspended in water, with a gavage. die route selected as the most efficient way to deliver an accurate dosage. Separate range-finding studies were conducted to select appropriate dosages for each study, HIS Study Designs All studies: Crl:CD(SD)IGS BR rats 90-Day Subchronic Toxicity Dosages: 0, 10,60 or 300 mg/kg/day (35% a.i, in IPA). 60 mg/kg/day IPA No. of animals: 10/sex/group -- main study, 10/sex/group intermediate, tow 1-montli recovery, 5/sex/group 3-month recovery Dosing Regimen: 90 days all groups, I-rnonth and 3-mondi recovery Parameters; U.S. EPA Health Effecis Test Guideline OPPTS 870,3100 Developmental Toxicity Dosages: 0.625. 1250, or 2500 mg/kg/day (20% a-i. in water) No. of animals: 22 pregnant females/group Dosing regimen: Gestation Days 6-20 Parameters: U.S. EPA Health Effects Tcsi Guidelines OPPTS 870.3700 Prenatal Developmental Toxicity Slady One-Generation Reproduction Dosages: 0. 75. 500. and 3500 mg/kg/day (20% a.i, in water) No. of animals: 20/sex/group Dosing Regimen: 70 days prior to mating and throughout mating -- females During gestation and laciaiion - dams only Parameters: Special Design (Fgurc I) males ani Figure 1 - Study Design I Subchronic Study Results la-Life Parameters No monality, clinical signs of toxicity, or behavioral abnormalities Significant lower body weight at 300 mg/kg/day - males only (Figure 2) Low food consumption and food efficiency at 300 ing/kg/day - ina!cs only Figure 2 Male Body Wtiglits Liver Effects Effects commonly observed in rats with many organofluoride materials, such as peroxisome proliferation, hepatomegaly, and increased liver weights were observed ill this study Increased hepatic B-oxidalion activity at 60 and 300 mg/kg/day - males and females (Figure 3) Elevated liver weights and hypertrophy at 60 and 300 mg/kg/day in males and at 300 mg/kg/day in females (Figure 3) Elevated liver enzymes al ail doses - males; at 300 mg/kg/day -- females Figure 3 Liver Effects Clinical Pathology Red cell mass parameters we (Table 1). These parameters affecied at 60 and 300 mg/kg/day - male rats only 'ere not affected in die reproduction study. Table 1 - Parameters of Red Cell Mass Dosaee (me/kss/da'f'): 10 60 300 Red Blood Cells Day 38 Day 93 3-nionili Recovery 101% 99% ND 97% 99% 97% ND 94% 96% 94% 90% 97% Hemoglobin Duy 38 Day 92 1-niiiiuli Recovery 3-liHMlll Recovery 39% 95% ND 97% 97% 95% ND 94% 93% 91% 94% 92% Heniarocril Day 38 Day 92 3-iiiuinli Rccuwiy 0% 90% ND 98% 98% 35% ND 34% 94% 91% 93% 93% jSy^Subchronic and Reproductive Study Results Hislopalhology In Ac subchronic study with active suspended in alcohol there were adverse liver tesions (focal necrosis) that did not occur in (he study with active ingredient suspended in water. Similarly, chronic progressive nephropathy was evident in the kidneys of animals dosed with active in water (reproductive study), but not in the subchronic study. Other histopathoiogical lesions were similar. EXB.EC 35 (^liv. ,, PA) Table 2 - Histopatiloiogy Effecis SBbdiniiilcAdiitI FuMte Lfvcr {^1TM(, NOB, PI Adill sp/re niyroidhyfnmph)1W focal necrojil Livo- hwenroiilif ' ' Kidassy fhioair pmensslvf hipcnmplnW li'llirnmsili\WFI HH|fDevelopmental Toxicity Results Maternal Toxicity * Reductions in body weight parameters al 1250 and 2500 mg/kg/day, (Table 3) Effects on food consumption at 2500 mg/kg/day. Fetal Toxkily * Reduced body weight parameters at 2500 mg/kg/day No malformations and no-compound-relaied variations at any cose Ie' Table 3 - Developmental Effects Maternal Eficcis Fetal EfTecli Mean body wcighl (l7) Adjuslsd liudy weiElH(i36t) Food consumption tiS'S') Mean body wciglil ti4l Mean liody wcielil (l5t) Adjusted body weight ti32t) No cITccIs | Reproduc Reduced body weigh females Reduced food consum Reduced body weigh males No effect on P] gener No effect on P| gener No effect on F] litter Reduced F] pup binh Do N M B Da Da Da Da Da Da Da Da Da } Summar Subchronic Study Based on liepatoc male rats ' Based on elevate 300 mi/kg/day, a Developmental Toxici Based on decreas for maternal effec Based on decreas was 1250 mg/kg/ Not a selective d One-Generation Repr Based on reducti period for F, litte was 5QO mg/kg/d and F[ adult male study- | Conclus The target organs of red blood cells The phosphate surfac The reproductive tox offspring growth imp VS Acknow We wish to iliank the f Nancy E. Everds for Linda A- Malley for ' John C, O'Connor fo Primary Technicians Gregory L. Poindext Poster Preparation-, M