Document 66RZNw6Y4yOJNX56ENp6XN1o

ARX&0357 CHRONIC TOXICITY TO FISH TEST SUBSTANCE Identity: N-ethylperfluorooctane sulfonamidoethanol; may also be referred to as N-EtFOSE Alcohol or FM-3422. (1-Octanesulfonamide, N-ethyl1 I 1I2 , 2 I3 ,3 )4 ,4 ,5 ,5 ,6 I6,7,7,8>8I8-heptadecafluoro-N-(2-hydroxyethyl)-, C A S# 1691-99-2) Remarks: Material is an off-white, waxy solid of uncharacterized purity. The laboratory conducting the study refers to N-EtFOSE Alcohol as METHOD: Method Followed: The methodology for the egg and fry exposure closely followed that presented in "Proposed recommended bioassay procedure for egg and fry stages of freshwater fish" (U.S. EPA, 1972). Type (test type): Flow-through GLP: No Year study performed: 1978 Species/Strain/Supplier: Fathead minnow (Pimephales promelas) eggs from the brood stock of the U.S. Environmental Protection Agency's Environmental Research Laboratory in Duluth, Minnesota. Analytical monitoring: Temperature, dissolved oxygen concentration, and pH were monitored daily, alternating between aquaria such that each aquarium was measured once each week. Remarks: One thousand ml water samples were taken from each aquarium at the initiation of the test (day 0), when hatching was completed (day 4), and weekly thereafter (days 11,18, 25, and 32) for determination of N-EtFOSE Alcohol concentration. Samples were taken with a beaker from a point approximately midway between the surface, bottom and sides of each aquarium, and stored in 1000 ml amber glass bottles with foil lined caps. All samples taken during the test were stored at room temperature and shipped on May 3 1 ,1 9 7 8 to the 3M Company. Exposure period: 4, 11, 18, 25, and 32 days Statistical Methods: Means of measured biological parameters from duplicate aquaria were subjected to analysis of variance (Steel and Torrie, 1960, completely randomized block design, P=0.05). Data for percentage survival and 006388 percentage hatch were transformed to arc sin square root of percentage prior to analysis. Test Condition Remarks: Exposures were initiated within 48-hours after egg fertilization and continued through 30 days post-hatch. The diluent water was well water pumped to a concrete reservoir where it was aerated before flowing to the exposure system through aged PVC pipe, th e pH of this water ranged from 7.0-7.2; total hardness, 31-38 mg/L as CaC03; alkalinity, 26-29 mg/L as C a C 0 3; and specific conductance, 149-170 umhos/cm. Five nominal concentrations of N-EtFOSE Alcohol, ranging from 1.3-20 pg/L, control water (well water), and control water containing solvent (Dimethylsulfoxide (DMSO) at 24 mg/L) were delivered to duplicate test aquaria. A 50 ml gas-tight syringe was used to deliver 46 pi of a 0.84 mg/ml stock solution of N-EtFOSE Alcohol in DMSO through a stainless steel needle and polyethylene tubing to the mixing chamber of the diluter. Each glass test aquarium measured 40 X 20 X 25 cm with a 19 cm high standpipe drain which maintained a constant test water volume of 15.2 liters. The diluter delivered 0.50 liters of test water to each aquarium 185 times per day, yielding a 90% test water replacement time of approximately 9 hours. The aquaria rested in a water bath containing circulating water heated by immersion coil heater and regulated by a mercury column thermoregulator designed to maintain the test water temperature at 25C. Upon arrival at E G & G , Bionomics, the eggs were allowed to acclimate from 17.5C to the test temperature of 25C over a period of two hours. Sixty eggs were then randomly distributed to each of 14 egg cups which were then dipped in a 60 mg/l malachite green solution for 15 seconds to prevent fungal growth. One egg cup was then suspended in each of the 14 test aquaria. Egg incubation cups were glass jars (5 cm O.D., 8 cm high) with 40 mesh Nitex*screen bottoms. An egg cup rocker arm apparatus, as described by Mount (1968), was used to gently oscillate the egg cups in the test water. Dead eggs were removed and counted daily until hatching was completed. Percentage hatch was calculated based on the number of live fry per aquarium after hatching was completed compared to the number of eggs per aquarium (60) at the initiation of the exposure. To initiate the 30-day fry exposure, forty fry were randomly selected from each egg cup and transferred to the respective aquaria. Upon transfer of fry to the aquaria, the fry were fed newly hatched San Francisco Bay variety brine shrimp nauplii, ad libitum, three times daily throughout the exposure period. The aquaria were brushed and siphoned to remove excess 006389 food and fecal material three times per week. At 30 days post-hatch, percentage survival, mean total length, and mean wet weight were determined for fry from each aquarium. The fry were measured individually to calculate a mean and standard deviation total length while each fry group (fry from one aquarium) was wet weighed to calculate a mean wet weight. At the termination of the test, the fry from the control and the high concentration (20 (j,g/L) were preserved in 10% buffered formalin while the fry from the other test aquaria were frozen. Ten formalin-preserved fry (5 from each replicate) from the control and the high concentration underwent histopathological examination of a transverse section of the nares and cephalic extension of the lateral line (See Table 3). The remaining preserved fry and frozen fry were analyzed at a later date (by 3M Company) for N-EtFOSE Alcohol concentrations. RESULTS______________________________________________________________ The biological data generated in this study indicate that hatchability of eggs, and percentage survival, mean total length and mean wet weight of fry were not adversely affected at any N-EtFOSE Alcohol concentration tested. Remarks: Water quality parameters measured during the egg and fry exposure exhibited little variation between test days and test chambers. Dissolved oxygen ranged from 8.2-10.2 mg/L. Temperature ranged from 23.5-26.5C and pH ranged from 6.9-7.5. 006390 TABLE 1 PERCENTAGE HATCH OF EGGS, PERCENTAGE SURVIVAL, MEAN AND STANDARD DEVIATION TOTAL LENGTH, AND MEAN WET WEIGHT OF FATHEAD MINNOW (Pimephales promelas) FRY EXPOSED TO N-ETFOSE ALCOHOL FOR 30 DAYS POST-HATCH. Nominal concentration (ng/i) Control Solvent Control 1.3 2.5 5.0 10 20 Replicate A B A B A B A B A B A B A B Hatch % Survival % 93 82 92 92 92 82 93 90 90 92 92 92 97 90 95 95 88 100 92 92 93 92 92 88 92 98 87 98 30 Days Mean length in mm (SD) 21(3) 21(2) 20(3) 20(4) 21(3) 21(3) 21(3) 21(2) 20(4) 21(2) 21(3) 23(2) 21(2) 21(2) Mean weight in mg 69 71 64 66 65 70 65 64 58 69 60 79 64 66 The 1000 ml water samples taken during the study (described in "Analytical Monitoring") were analyzed at 3M Company. Results indicated excellent recovery. See Table 2. 006391 TABLE 2 ANALYSES FOR N-ETFOSE ALCOHOL IN BIONOMICS SAMPLES TO VERIFY RECOVERY CAPABILITIES Theoretical Cone, Sample I.D. (ppb) A5619 Control A A5620 Control B A5621 Solvent Control A A5622 Solvent Control B A5617 1.25 A5618 1.25 A5670 1.25 A5670 1.25 A5615 2.5 A5616 2.5 A5613 5 A5614 5 A5611 10 A5612 10 A5609 20 A5610 20 (a) received broken in shipment (b) replaces original samples A5617; A5618 Determined (ppb) <0.1 <0.1 <0.1 <0.1 (a) (a) 1.1; 1.0(b) 1.0; 1.6(b) 2.5; 2.6 1.2; 1.3 5.2; 5.2 4.8; 5.1 10.2; 9.9 9.7; 10.1 20.8; 19.8 19.8; 20.6 TABLE 3 RESULTS OF HISTOPATHOLOGICAL EXAMINATION OF FRY OF FATHEAD MINNOW (PIMEPHALES PROMELAS) EXPOSED TO 20 i^g/l OF N-ETFOSE ALCOHOL3 Test Material Number of Histopathological Observations Findings Control 10 3/10 Normal 6/10 Liver Fatty Change 3/10 Gill Hyperplasia (Epithelium) I N-EtFOSE Alcohol 10 9/10 Normal 1/10 Gill Hyperplasia a. Work performed under contract to EG & G Bionomics Laboratory NOTE: "Only those tissues which were missing or contained demonstrable change are listed. The only tissue changes observed were hyperplasia of gill lamellar epithelium and fatty change of the liver. These changes were judged to be minimal and consistent with changes seen routinely in healthy fish. Autolipis of gill tissue was observed in several fish. This change was probably due to the poor penetration of the buffered formalin to the posterior dorsal portion of the gill space." 006392 CONCLUSIONS The No Effect Concentration (NOEC) of N-EtFOSE alcohol for hatchability, growth, histopathology and survival of Fathead minnow fry is greater than 19.820.8 yg/L measured (20 yg/L nominal), the highest concentration tested. DATA QUALITY______________________________________________________ _ Reliability: Klimisch ranking = 2. This study meets all the criteria for quality testing, but has several deficiencies. It lacks information on purity of the test substance, the production lot number from which the test sample was taken, and was not done at the reported solubility limit for the test substance. REFERENCES_______________________________________ _________________ This study was conducted by E G & G, Bionomics, Wareham, Massachusetts, 1978 on the request of 3M Company. Research Report "THE EFFECTS OF CONTINUOUS AQUEOUS EXPOSURE TO 78.01 (N-ETFOSE ALCOHOL) ON HATCHABILITY OF EGGS AND GROWTH AND SURVIVAL OF FRY OF FATHEAD MINNOW (Pimephales promelas)." Report # BW-78-6-195, E G & G, Bionomics, Aquatic Toxicology Laboratory, 790 Main Street, Wareham, Massachusetts, June 1978. Research Report "SUMMARY OF HISTOPATHOLOGICAL EXAMINATION OF FATHEAD MINNOW (Pimephales promelas) EXPOSED TO 78.01 (N-EtFOSE ALCOHOL) FOR 30 DAYS." Report # BW-78-6-195, E G & G, Bionomics, Aquatic Toxicology Laboratory, 790 Main Street, Wareham, Massachusetts, June 1978. 3M Technical Report Summary, "Analytical Methodology and Support", Fate of Fluorochemicals, Project Number 9970612643, Report Number 008, Arthur Mendel, January 17,1979 O T H E R __________________________________________________________ Submitter: 3M Company, Environmental Laboratory, P.O. Box 33331, St. Paul, Minnesota, 55133 Last changed: 5/18/00 006393