Test CAS RN Common Structure % CAS Name

15-25

Category Sub-Category Test Test Substance Report Result Summary

Mammalian Toxicity Acute Oral 

### ALD (rats) = > 5000 mg/kg 1988c).

A single dose of suspended in corn oil was administered by oral gavage to one male rat per dose level. Dose levels were 450, 670, 1000, 1500, 2300, 3400, 5000, 7500, and 11,000 mg/kg. Following administration of the test material, rats were observed for clinical signs of toxicity. Surviving rats were weighed and observed daily until signs of toxicity subsided, and then at least 3 times per week for 14 days after dosing. Observations for mortality were made daily throughout the study.

The lowest dose of test material which resulted in death was 2300 mg/kg. Deaths occurred up to 5 days after dosing. The animal dosed at 3400 mg survived. Animals dosed at 5000 mg/kg and greater died. The survival of the animal dosed at 3400 mg/kg was attributed to animal variability, and did not change the ALD. The only clinical sign of toxicity observed at nonlethal doses was yellow-stained perineum in the rat dosed at 3400 mg/kg 4 days after dosing. Moderate to severe weight losses were observed in all rats up to 4 days after dosing.

Under conditions of this study, the ALD for was 2300 mg/kg. This material was considered to be slightly toxic when administered as a single oral dose to male rats 1988c).

Mammalian Toxicity Acute Dermal 

### was evaluated for skin irritation potential
in New Zealand White rabbits. A 0.5 g aliquot of the test substance, moistened with dimethyl phthalate, was applied directly to a 2-inch gauze square that was then placed on the shaved test site. The patch was held in place with tape. Rubber sheeting was then wrapped around the animal and secured for an occlusive dressing. The rabbits were exposed to the test substance for 24 hours after which the test substance was removed by gentle washing with warm water. The test substance was moistened with dimethyl phthalate, but did not dissolve. Therefore, some of the compound may have fallen from the test site during the exposure period. Approximately 24 and 48 hours after application of the test material, the test sites were evaluated for erythema, edema and other evidence of dermal effects and were scored according the Draize scale. The adjacent areas of the untreated skin were used for comparison.

No erythema or edema was observed in any of the treated rabbits. Under conditions of this study, was not a skin irritant ( 1988b). Test Result Summary Toxicology Summary The toxicological information summarized below is NOT on the PMN substance, but on a related substance that may be of interest to the Agency. Full study reports can be provided upon request.Mammalian Toxicity Acute Eye 

### was evaluated for acute eye irritation potential in 2 male New Zealand White rabbits. A 0.01 g aliquot of test substance was administered to 1 eye of each animal. Both eyes of 1 animal remained unwashed. Approximately 20 seconds after the test material was administered, both eyes of the remaining animal were rinsed for 1 minute with lukewarm tap water. The conjunctiva, iris, and cornea of each eye were evaluated for evidence of eye irritation approximately 1 and 4 hours and 1, 2 and 3 days following administration of the test substance. At each observation, the treated eyes were examined using illumination and magnification and scored according to the Draize scale. The untreated eye of each animal was also examined and used for comparison. Fluorescein dye was used to evaluate corneal ulceration at the 1-, 2-, and 3-day observation. Biomicroscopic examinations for corneal injury were conducted at the 1-, 2-, and 3-day observation and scored according to a numerical scale.

produced no irritation in either treated rabbit eye. Fluorescein stain and biomicroscopic examinations were negative for corneal injury in both treated eyes. Under the condition of this study, was not an eye irritant ( 1988a).

Ecotoxicology Aquatic Studies Fish 

### 96-hour LC50 (fathead minnow, Pimephales promelas) = > 1000 mg/L ( 1989).

The acute toxicity of to the fathead minnow, Pimephales promelas was determined in an unaerated, 96-hour, static test. The study was conducted with 3 concentrations of test material and a dilution water control at a temperature of 21.7 - 22.6°C. Exposure of fathead
minnows to the dilution water control and nominal test material concentrations of 360, 600, and 1000 mg/L resulted in no mortalities at the end of 96 hours. All test solutions (from 360 to 1000 mg/L) were milky with undissolved test material floating on the surface and settling to the bottom. The solubility of was less than 360 mg/L, as there was
undissolved test material in all test concentrations. was found to exhibit no acute toxicity at test concentrations above its solubility in water ( 1989).

** Toxicological Studies also available for

1.

2.
3.

4.CAS RN Common Name Structure % CA Index Name 59587-38-1 Perfluorohexylethylsu
lfonic acid F(CF2) 6CH2CH2SO3

- K+ 100 1-Octanesulfonic acid, 3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluoro, potassium salt (1:1)

Category Sub-Category Test Result Summary Mammalian Toxicity Acute Oral 

### LD50 (rat; Probit method) = 2000 mg/kg

LD50 (rat; Arcsinus method) = 2400 mg/kg

5 male and 5 female rats per group were administered 0.5, 1, 2, or 4 g/kg of the test substance ( tested in solid form) in olive oil. They were observed during the first hours after intubation and then for 14 days. Clinical signs of toxicity observed included nasal and/or eye congestion and polyuria (>500 mg/kg), red-stained fur (>1000 mg/kg), ptosis (1000 and 2000 mg/kg), apathy (2000 mg/kg), prostration (>2000 mg/kg), difficult locomotion (2000 mg/kg), diarrhea (4000 mg/kg), weight loss (>2000 mg/kg), moribund (4000 mg/kg), and piloerection (4000 mg/kg) ( 1977a).

Mammalian Toxicity Acute Dermal 

### The test substance ( tested in solid form) was not a skin
irritant in rabbits.

The test substance (0.3 g) was applied to the surface of the scarified or non scarified skin of 6 male New Zealand white rabbits via gauze squares. The gauze squares were held in place with an occlusive adhesive patch. Then tape was wound around the rabbit to complete the fixing of the patches. The rabbits were then placed in restraining devices for 24 hours. After 23 hours, the patches were removed, and 1 hour later the primary irritation index was evaluated. Another reading was made 48 hours later. The primary cutaneous irritation score was 0 ( 1977a).

Mammalian Toxicity Acute Eye 

### The test substance ( tested in solid form) was classified as very irritating to the rabbit eye.

Six male New Zealand white rabbits were tested. The test substance (0.1 g) was introduced into the rabbit’s left eye, and the right eye served as a control. The eyelid was held closed for several seconds, giving a slight massage to avoid loss of the substance. The eyes were not washed after administration. Evaluations were performed at 1 and 24 hours, and at 2, 3, 4, and 7 days after administration. The index of acute ocular irritation (IOA) was 15.67 and the average index of ocular irritation after 4 days (IOM) was 3.33. The test substance was classified as very irritating to the rabbit eye ( 1977a). The toxicological information summarized below is for PMN Substance #2

Toxicology Summary

(Test Substance)Mammalian Toxicity Subchronic Oral 

### A biopersistence screening 10-dose oral gavage study in rats with a recovery period was conducted. Two groups of 5 male rats each were given 25 mg/kg/day of the test substance ( tested in solid form) for 10 consecutive days. Approximately two hours after dosing on test days 1 and 5, blood was collected from the orbital sinus of each rat from one five-dose group. Approximately 2 hours after the last dose, these rats were euthanized and blood, livers, and fat were collected. Blood was collected from the remaining five rats on test days 13, 24, and 52. These rats were euthanized on test day 94, and blood, livers, and fat were collected. Body weight and clinical signs were recorded on each dosing day, and then weekly during the recovery period. On selected days (5, 10, 13, 24, 52, and 94), five rats per group were euthanized and the blood and livers collected. Body weights and clinical signs were recorded on each day of dosing and then approximately every other week during the recovery period. Additionally, one negative control, deionized water, and two positive controls were
tested.

No deaths occurred during the study. No compound-related clinical signs of toxicity were observed in the rats dosed with the test substance. The mean body weights and mean body weight gains of the test rats were comparable to the negative controls and equal to or greater than the positive controls. The mean relative liver weights of rats dosed with the test substance was 12% higher than the negative control at day 10 and 6% higher at day 94. The mean relative liver weights of rats dosed with the positive control materials were 38 and 88% higher on test day 10, but were comparable to control on test day 94. The increased liver weights appear to be associated with increased liver concentrations of fluorine equivalents of test or positive control substances, particularly on day 10.

The test substance normalized μm equivalents in rat blood increased during the dosing period and did not appear to reach steady-state. The
area under the curve (estimated to infinity)was calculated and normalized for fluorine content for the test substance and each positive control and further normalized to dose. The AUCINF/D for the fluorine component of the test substance was 17,865 compared to AUCINF/D values of 522,474 and 81,507 for the positive controls.

The concentrations in fluorine in the livers from rats dosed with the test substance was 423.64 μM equivalents on day 10. On day 10, the mean μM equivalent concentrations of fluorine in the fat from rats dosed with the positive control materials were approximately 24-fold and 7-fold greater than the fluorine concentration in fat from rats treated with the test substance. By day 94, excess fluorine was below the limit of quantification in fat of rats dosed with the test substance or one positive
control. Some retention of fluorine was evident in the fat from rats dosed with the other positive control material.

The mean μM equivalents of fluorine in the liver from animals dosed with the test substance were higher than levels in the blood and fat.

Under the conditions of this study, absorption of the test substance by rats following oral gavage administration was evident based on quantification of fluorine in the blood, liver, and fat. A very minimal increase in relative liver weights was observed for the test substance. Changes in relative liver weights were greater for the positive controls, and were associated with greater concentrations of fluorine in the liver particularly at day 10. The tissue concentrations declined after cessation of dosing with some retention of fluorine in the blood and liver based on quantification on the end (day 94) of the study. However, fluorine concentrations and the AUCINF/D were significantly lower than those in rats dosed with the positive control materials ( 2008a).Ecotoxicology Environmental
Fate Bioaccumulation/

Bioconcentration 

### An in vitro trout hepatocyte bioaccumulation screen was performed with the test substance ( tested in solid form). Two experiments with the test substance and 1 experiment with each control was performed. Two replicates per experiment for the test, positive control (4-nonylphenol), and dead cell control were performed. The cell concentration was 2x106 cells/mL, in a dose vehicle of acetonitrile, at a pH of 7.8, incubation temperature of 10-12°C, and final concentration of 5 μM (2.33 ppm). The time points examined were 5, 15, 30, 60, 120, and
240 minutes.

No apparent loss of the parent compound was observed within 4 hours of incubation. These results suggest that the bioaccumulation potential of this test substance would not be significantly affected by fish metabolism ( 2007i).

Ecotoxicology Environmental
Fate Bioaccumulation /

Bioconcentration 

### Rainbow trout, Oncorhynchus mykiss, were exposed to under flow-through conditions for 56 days during the uptake (exposure) phase of the study followed by a 28-day depuration phase in clean dilution water. Two nominal aqueous test substance concentrations (1 and 10 μg/L), a single nominal spiked diet concentration (10 μg/kg), and a dilution water control were used for testing with 60 fish per exposure treatment and control. An error in preparation of the aqueous test concentrations and delay in analytical confirmation of the test substance concentrations resulted in the day 1-13 uptake phase aqueous test substance concentrations being below the analytical method limit of detection of 0.06 μg/L. This error was corrected upon discovery as reflected in the aqueous test substance concentrations for study days 14 through 56.

The mean, measured concentrations of in the aqueous test solutions from days14-56 were 1.08 μg/L and 9.32 μg/L and the measured concentration in the spiked trout chow diet used for the duration of the dietary uptake phase of the study was 13.1 μg/Kg. One day after initiation of the depuration phase there were no detectable levels of present in the aqueous test solution test chambers.
The dilution water control and the control trout chow diet contained no
detectable concentrations of during the study. Fish tissue
residue concentrations of reached steady-state equilibrium within 42 days or earlier in both aqueous and dietary exposures. The k1 values (uptake rate constant) for days 14-56 of the aqueous exposures in whole fish based on measured tissue residues were 4.462 and 0.452 per day for the nominal 1 and 10 μg/L nominal aqueous test concentrations while the day 14-56 depuration rate constants, k2, in whole fish based on measured tissue residues were 0.125 and 0.177 per day, respectively. After a maximum of 24 days of depuration, the measured concentrations in the whole fish were approximately 95% or less of the whole fish tissue concentrations at the end of the uptake phase. However, measured concentrations in whole fish at the end of the uptake phase on day 56 were already less than 50% of the maximum uptake phase concentration in whole fish for both aqueous test concentrations and the dietary exposure concentration of The steady-state bioconcentration factors (BCF) for the nominal 1 and 10 μg/L aqueous test substance concentrations were 22 and 3, respectively. The respective kinetic bioconcentration factors were 36 and 3, respectively, for the nominal 1 and 10 μg/L aqueous test substance concentrations. The dietary assimilation efficiency for the growth corrected half-life, and the dietary biomagnification factor (BMF) for the dietary exposure were 0.435, 23.5 days, and 0.295, respectively. All test acceptance criteria were met ( 2009b).Ecotoxicology Aquatic Studies Fish 

### 96-hour LC50 (rainbow trout, Oncorhynchus mykiss) >107 mg/L.

The acute toxicity of the test substance ( tested in solid form) to rainbow trout was determined in an unaerated, 96-hour toxicity test following OECD Guideline 203. Nominal concentrations tested were 0, 7.5, 15, 30, 60, and 120 mg/L. The mean water temperature was 12.1°C (range of 11.9 12.3°C). One test chamber was used per test concentration with 10 fish in each chamber. Control fish ranged from 3.0 to 3.3 cm in standard length and 0.260 to 0.352 g in wet weight, blotted dry, at test end. Dilution water loading at test end was
0.3041 g/L. Mean measured concentrations were none detected (limit of detection 0.0008 mg/L), 6.82, 14.6, 27.7, 53.4, and 107 mg/L for the 0, 7.5, 15, 30, 60, and 120 mg/L concentrations, respectively. No mortality was observed at any concentration. Dilution water quality was acceptable based on OECD criteria and specifications. All chemical and physical parameters were within the expected ranges. Total alkalinity, EDTA hardness, and conductivity of the dilution water control and highest test substance concentration at test start ranged from 48 to 49 mg/L as CaCO3, 133 to 137 mg/L as CaCO3, and 230 to 255 μmhos/cm, respectively. During the test, dissolved oxygen concentrations ranged from 8.5 to 10.4 mg/L, pH ranged from 7.4 to 7.6, and mean temperature was 12.1°C with a range of 11.9 to 12.3°C ( 2007e).

Ecotoxicology 

### 90-day early life-stage toxicity NOEC (rainbow trout, Oncorhynchus mykiss) = 2.62 mg/L.

The effect of the test substance ( tested in solid form) on hatching, growth, and survival of rainbow trout embryos, alevins,and fingerlings were assessed in an intermittent renewal 90-day early lifestage toxicity test in accordance with OECD Guideline 201 and US EPA OPPTS 850.1400. Nominal test concentrations were 0, 0.625, 1.25, 2.5, 5, and 10 mg/L. A total of 80 embryos were exposed per concentration (20 embryos per embryo cup, 2 cups per replicate, 2 replicates per concentration) at test start. On day 40, after swim-up had begun in the control, the fingerlings were thinned to a total of 30 fish per concentration (15 fish per replicate, 2 replicates per concentration). Analytical
verification of test substance concentrations were made on test solutions sampled on day -3, day 0, once weekly, and on day 90 at test end. Test solutions were supplied to each replicate test chamber at a rate of approximately 1.25 L of test solution per test chamber each hour, resulting in approximately 5 test solution volume additions of 6 L (30 L total) every 24 hours. Embryos and alevins were held in relative darkness until day 39, and then held under a photoperiod of 16 hours light and 8 hours darkness (which included 30 minutes of transitional light between light and dark intervals) for the remainder of the study. Test solutions were maintained between 11.9 and 12.9°C (mean 12.2°C). Daily observations were made for assessment of number of dead eggs at hatching, first and last day of hatching, first day of swim up, survival and abnormalities from hatching to thinning, and survival and abnormalities from thinning to test end. Standard length and blotted wet weight of surviving fingerlings were determined at test end.

Mean measured test concentrations were 0, 0.857, 1.66, 2.62, 4.85, and 8.70 mg/L at the 0, 0.625, 1.25, 2.5, 5, and 10 mg/L concentrations, respectively. The NOEC was 2.62 mg/l based on mean, measured concentrations and first day of hatching values. The LOEC (lowest observed effect concentration) and MATC (maximum acceptable toxicant concentration) for the same endpoint were 4.85 and 3.56, respectively. The NOEC values were 4.85 mg/L for percent eggs hatching and first day of swimup and >8.7 for the other test endpoints ( 2007f). Fish Aquatic StudiesEcotoxicology Aquatic Studies Invertebrates 

### The acute toxicity of the test substance ( tested in solid form) to unfed Daphnia magna neonates, less than 24 hours old at test start was determined in an unaerated, 48 hour toxicity test following OECD Guideline 202. Nominal concentrations tested were 0, 7.5, 15, 30, 60, and 120 mg/L. The mean water temperature was 19.9°C (range of 19.5-20.1°C). Four replicates with five daphnids per replicate were used per test substance concentration and control. Mean measured concentrations were none detected (limit of detection 0.01 mg/L), 7.09, 14.3, 27.8, 53.4, and 109 mg/L for the 0, 7.5, 15, 30, 60, and 120 mg/L concentrations, respectively. Immobility at the end of 48 hours was 0, 25, 15, 25, 30, and 50% at 0, 7.09, 14.3, 27.8, 53.4, and 109 mg/L, respectively. Dilution water quality was acceptable based on OECD criteria and specifications. All chemical and physical parameters were within the expected ranges. Total alkalinity, EDTA hardness, and conductivity of the dilution water control and highest test substance concentration at test start ranged from 50 to 52 mg/L as CaCO3, 134 to 136 mg/L as CaCO3, and 280 to 315 μmhos/cm, respectively. During the test, dissolved oxygen concentrations ranged from 7.5 to 8.7 mg/L, pH ranged from 7.6 to 8.1, and mean temperature was 19.9°C with a range of 19.5 to 20.1°C ( 2007g).

### 72-hour EbC50 (green alga, Pseudokirchneriella subcapitata) > 96 mg/L (cell count and area under the curve).

0-72-hour EbC50 (green alga, Pseudokirchneriella subcapitata) > 96 mg/L (area under the curve).

72-hour ErC50 (green alga, Pseudokirchneriella subcapitata) > 96 mg/L (growth rate).

The toxicity of the test substance ( tested a solid form) to the green algae was determined in a 72 hour, static toxicity test following OECD Guideline 201. The study was conducted with a blank control, an abiotic control, and 5 nominal concentrations of the test substance (6.25, 12.5, 25, 50, and 100 mg/L) at a mean lighting intensity of 8498 lux (range of 8200 to 8670 lux), a mean temperature of 23.5°C(range of 23.5-23.6°C), and a shaking speed of 104 rpm. Synthetic algalassay-procedure (AAP) nutrient medium was used as the test diluent and blank control. Test solutions were not renewed. Three replicates were used per test concentration and 6 replicates were used for the blank control. A single test flask was used for the abiotic (stability) control. Healthy cell count, area under the growth curve, and growth rate were
determined at 24-hour intervals over the 72-hour test.

Mean measured concentrations were none detected (limit of detection 0.006 mg/L), 6.20, 12.4, 24.2, 47.6, 96, and 99.3 mg/L for the blank control, 6.25, 12.5, 25, 50, 100, and abiotic control (100 mg/L) concentrations, respectively. Inhibition of growth expressed as biomass (cell number) of Pseudokirchneriella subcapitata exposed to nominal concentrations of 6.25, 12.5, 25, 50, and 100 mg/L for 72 hours was -8, 2, 14, 3, and 20%, respectively. Inhibition of growth expressed as area under the growth curve was -9, 4, 19, -1, and 14%, respectively. Inhibition of growth expressed as the average specific growth rate was -1, 0, 2, 0, and 4%, respectively. Healthy cell counts increased in the blank control by at least a factor of 16 in 72 hours and the coefficient of variation of average specific growth rates during the whole test period (0- 72 hour) in replicate control cultures did not exceed 7%, thereby satisfying the appropriate test acceptance criteria ( 2007h). Ecotoxicology Aquatic Studies OtherTest Substance CAS RN Common Name Structure % CA Index Name

35.6

30

N/A

30

>99

100% (based on testing done on solid
form)
N/A

Category Sub-Category Test Test Substance Report Number Result Summary

Mammalian Toxicity Acute Oral LD50 (rat; Probit method) = 1871 mg/kg

LD50 (rat; Litchfield and Wilcoxon) = 1871 mg/kg

LD50 (rat; Arcsinus method) = 2000 mg/kg

5 male and 5 female rats per group were administered 0.5, 1, 2, or 4 g/kg of the test substance ( tested in solid form) in olive oil. They were observed during the first hours after intubation and then for 14 days. Clinical signs of toxicity observed included red-stained fur (>1000 mg/kg), prostration (>1000 mg/kg), nasal congestion (2000 mg/kg), moribund (4000 mg/kg), and piloerection (4000 mg/kg) ( 1977b).

Mammalian Toxicity Acute Dermal The test substance ( tested in solid form) was a slight skin
irritant in rabbits.

Undiluted test substance (0.3 g moistened with 0.5 mL water) was applied to
the surface of the scarified or non-scarified skin of 6 male New Zealand white rabbits via gauze squares. The gauze squares were held in place with an occlusive adhesive patch. Then tape was wound around the rabbit to complete the fixing of the patches. The rabbits were then placed in restraining devices for 24 hours. After 23 hours, the patches were removed, and 1 hour later the primary irritation index was evaluated. Another reading was made 48 hours later. The primary cutaneous irritation score was 0.71 ( 1977b). The toxicological information summarized below is NOT on the PMN substance, but on a related substance that may be of interest to the Agency. Full study reports are submitted with this PMN. Publically available literature studies will not be submitted.

(Test Substance)

Toxicology Summary *STN's CAS Registry Database has given a CAS RN to a product name. Test substance is CAS RNThe test substance ( tested in solution) was not a dermal sensitizer in the local lymph node (LLNA) assay in mice.

Five groups of 5 female CBA/JHsd mice were dosed for 3 consecutive days with 0, 5, 25, 50, or 75% of the test substance on both ears following OECD and US EPA OPPTS 870.2600 guidelines. N,N-dimethylformamide (DMF) was the diluting vehicle. One group of 5 female mice was dosed for 3 consecutive days with 25% hexylcinnamaldehyde (HCA) in DMF as a positive control. On test day 5, mice received 3H-thymidine by tail vein injection and were sacrificed approximately 5 hours later. The cell proliferation in the draining auricular lymph nodes of the ears was evaluated.

No differences in mean body weights or body weight gains were observed. No clinical signs of toxicity were observed. Statistically significant increases in cell proliferation measurements compared to control were observed at the 25, 50, and 75% test concentrations. Stimulation indices (SIs) of less than 3.0 were observed at all test concentrations. Therefore, the EC3 value (the estimated concentration required to induce a threshold positive response, i.e., SI = 3) for the test substance under the conditions of this study was not calculable. A 25% concentration of the positive control produced a dermal sensitization response in mice. Therefore, the test substance did not produce a dermal sensitization response in mice ( 2007a). Mammalian Toxicity Acute Dermal

Mammalian Toxicity Acute The test substance ( tested in solid form) produced irritation with necrosis of the epithelium in the rabbit eye.

Six male New Zealand white rabbits were tested. The test substance (0.1 g) was introduced into the rabbit’s left eye, and the right eye served as a control. The eyelid was held closed for several seconds, giving a slight massage to avoid loss of the substance. The eyes were not washed after administration. Macroscopic examination of the eyes was impossible because of the presence of large edemas of the palpebral conjunctiva. Therefore, the treated eyes of 2 rabbits per time point were removed at 1, 24, and 48 hours, and were fixed and examined microscopically. Marked irritation was observed on the level of the cornea after application of 0.1 g of the test substance to the surface of the eye. Necrosis followed by disappearance of the epithelial surface was observed and was accompanied by alteration in the corneal stroma. At the level of the stroma there was pyctonic cores of fibroblastic cells and polynuclear infiltrates. Irritation increased over time, as the test substance penetrated more deeply into the corneal stroma ( 1977b). EyeMammalian Toxicity Subchronic Oral The test substance ( was incorporated into nutritionally adequate chow and fed to male Crl:CD-1®BR mice for 14 consecutive days. Diets contained 0, 3, 30, 300, or 3000 ppm of the test substance. Mice were weighed and observed for clinical signs of toxicity once each week. At the end of the two-week feeding period, all mice were euthanized and necropsied. The livers were weighed, and liver weight/final body weight ratios were
calculated.

No deaths occurred during the study. Other than a body weight effect in mice from the 3000 ppm dose group, no adverse clinical signs of toxicity were observed during the study. Significant increases in mean absolute and relative liver weights were observed in mice from the 300 and 3000 ppm dose

groups.

The no-observable-adverse-effect level (NOAEL) was determined to be 30 ppm based on the increases in mean absolute and relative liver weight observed in the 300 and 3000 ppm dose groups and the decreases in body weight and body weight gains observed in the 3000 ppm dose group ( 1995c).

Mammalian Toxicity Subchronic Oral Groups of 20 male rats (Crl:CD®BR) were fed diets containing 0, 30, or 300 ppm of the test substance ( for 14 consecutive days. Rats were sacrificed at the end of the feeding period and on recovery days seven, 28, and 56. Livers were removed and weighed. Blood samples were taken for fluoride concentration analysis from rats sacrificed at the end of the feeding period and on recovery day seven.

No mortalities occurred during the study. No clinical signs of toxicity were observed. Significant increases in mean liver weights were observed in rats from the 30 and 300 ppm groups sacrificed at the end of the feeding period. No other liver effects were observed in these rats during the study.

Mean blood fluoride concentrations of rats sacrificed on recovery day 0 were 0.30 ppm in control rats, 5.30 ppm in the 30 ppm rats, and 8.19 ppm in the 300 ppm rats. On recovery day seven, mean blood fluoride concentrations were 0.94 ppm in control rats, 0.58 ppm in the 30 ppm rats, and 0.56 ppm in the 300 ppm rats ( 1995d).

Mammalian Toxicity Subchronic Dermal Three male rabbits were exposed dermally to the test substance ( tested in solution) for 90 days with a 7-day recovery period at a dose of 2000 mg/kg (30% aqueous solution). The test substance was applied daily (5 days out of 7) for 13 weeks on the shaved skin of the whole of the right flank
of each rabbit. The left flank served as a control. The test substance was applied and given a light massage of about 30 seconds to ensure maximal penetration. After the massage, excess material that had not penetrated the skin was wiped off with gauze. Body weights were recorded weekly. The skin was examined daily before application of the test substance, and a mean index of irritation was determined each week for each animal. After 90 days of repeated application, two samples of skin were removed by sterile biopsy and evaluated histologically. No effects were observed upon macroscopic or microscopic examination. No evidence of allergic reaction was observed. The average index of irritation for the 3 rabbits was 0.17 in the 1st, 5th, 6th, and 8th weeks. All other time points, the average index of irritation was 0. Under the conditions of this study, it was concluded that repeated dermal application of a 30% aqueous solution of the test substance for 90 days was well tolerated by rabbits ( 1977b).Mammalian Toxicity Gentic Toxicity In Vitro The test substance ( tested in solution) was not mutagenic in the bacterial reverse mutation test with Salmonella typhimurium.

A preliminary toxicity test was performed to define the dose levels to be used for the mutagenicity study. The test substance was then tested in two independent experiments, with and without a metabolic activation system, S9 mix, prepared from a liver microsomal fraction (S9 fraction) of rats induced with Aroclor 1254. Both experiments were performed according to the direct plate incorporation method except for the second test with S9 mix, which was performed according to the preincubation method. Five strains of Salmonella typhimurium (TA1535, TA1537, TA98, TA100, and TA102) were used. Each strain was exposed to five dose levels (three plates per dose). After 48-72 hours of incubation at 37°C, the revertant colonies were scored. The evaluation of the toxicity was performed on the basis of the observation of the decrease in the number of revertant colonies and/or a thinning of the bacterial
lawn. The test substance was dissolved in distilled water. Sodium azide (NaN3), 9-aminoacridine (9AA), 2-nitrofluorene (2NF), mitomycin C (MMC), and 2-anthramine (2AM) were used as positive controls.

The number of revertants for the vehicle and positive controls was as specified in the acceptance criteria. Since the test substance was freely soluble and almost non toxic in the preliminary test, the highest dose for the main test was 5000 μg/plate, according to the criteria specified in the international guidelines. The selected treatment levels were 312.5, 625, 1250, 2500, and 5000 μg/plate, for both mutagenicity experiments with and without S9 mix. No toxicity was observed, both with and without S9 mix. No increase in the number of revertants, both with and without S9 mix, in any of the five strains was observed. Therefore, under the conditions of this study, the test substance does not show any mutagenic activity in the bacterial reverse mutation test with Salmonella typhimurium, with and without S9 mix up to 5000 μg/plate as active material ( 2002).In Vitro Mammalian Toxicity Gentic Toxicity The test substance ( tested in solution) was positive for the induction of structural chromosome aberrations and negative for the induction of
numerical chromosome aberrations in CHO cells in both non-activated and S9 activated test systems, following OECD Guideline 473, EC Commission Directive 2000/32/EC Annex 4A-B10 no. L 136, and US EPA OPPTS Guideline 870.5375. A preliminary toxicity test was performed to establish the dose range for the chromosome aberration assay. The chromosome aberration assay was used to evaluate the clastogenic potential of the test substance.

Water was used as the test substance solvent based on the solubility of the test substance and compatibility with target cells. The test substance was soluble in water at a concentration of approximately 50 mg/mL, the maximum concentration tested for solubility.

In the preliminary toxicity assay, the maximum dose tested was 4280 μg/mL (10 mM). The test substance was soluble in water at all concentrations tested. Visible precipitate was observed in the treatment medium at dose levels >1284 μg/mL and dose levels >428 μg/mL were soluble in the treatment medium at the beginning and conclusion of the treatment period. Selection of dose levels for the chromosome aberration assay was based on cell growth inhibition relative to the solvent control. Substantial toxicity (i.e., at least 50% cell growth inhibition, relative to the solvent control) was observed at dose levels >428 μg/mL in all three treatment groups. Based on these findings, the doses chosen for the chromosome aberration assay ranged from 12.5 to 300 μg/mL for all three treatment groups.

In the initial chromosome aberration assay, the cells were treated for 4 and 20 hours in the non-activated test system and for 4 hours in the S9-activated test system. All
cells were harvested 20 hours after treatment initiation. The test substance was

soluble in water and in the treatment medium at all dose levels tested at the beginning and conclusion of the treatment period. Selection of doses for microscopic analysis was based on toxicity (the lowest dose with at least 50% reduction in cellgrowth and the next two lower doses) in the S9-activated 4-hour and the nonactivated 20-hour exposure groups.

The percentage of cells with structural aberrations in the S9-activated 4-hour exposure group was statistically increased (21.0%) above that of the solvent control at 300 μg/mL (p<0.01, Fisher's Exact test). The Cochran-Armitage test was also positive for a dose response (p<0.05). The percentage of cells with numerical aberrations was statistically increased above that of the solvent control at dose levels 50 and 150 μg/mL (p<0.01 and p<0.05, respectively, Fisher's Exact test). However, the percentages of cells with numerical aberrations at 50 and 150 μg/mL (6.5% and 6.0%, respectively) were within the historical solvent control range of 0.0% to 10.0%. Therefore, they ware not considered to be biologically significant. The percentage of cells with structural or numerical aberrations in the non-activated 20-hour exposure group was not significantly increased above that of the solvent control at any dose level (p>0.05, Fisher's Exact test).

Due to lack of a dose level with close to 50% reduction in cell growth or mitotic index, the chromosome aberration assay was repeated in the non-activated 4-hour exposure group. Although the dose levels selected for the repeat assay were 25, 100, 300, 325, 350, 375, 400 and 415 μg/mL, the repeat assay was inadvertently tested at the same doses used in the initial assay (12.5, 25, 50, 100, 150, 200, 250 and 300 μg/mL). Since the toxicity data from the repeat assay showed a shift in toxicity (50% reduction in cell growth and mitotic index was achieved), the study director concluded that the alternate doses tested in the repeat assay had no adverse effect on the outcome of the study. In the repeat chromosome aberration assay, the cells were treated for 4 hours in the non-activated test system. All cells
were harvested 20 hours after treatment initiation. The test substance was soluble in water and in the treatment medium at all dose levels tested at the beginning and conclusion of the treatment period. Selection of doses for microscopic analysis was based on toxicity (the lowest dose with at least 50% reduction in cell growth and the next two lower doses) in the non-activated 4-hour exposure group.

The percentage of cells with structural aberrations in the non-activated 4-hour exposure group was statistically increased (23.0%) above that of the solvent control at 300 μg/mL (p<0.01, Fisher's Exact test). The Cochran-Armitage test was also positive for a dose response (p<0.05). The percentage of cells with numerical aberrations in the test substance-treated group was not significantly increased above that of the solvent control at any dose level (p>0.05, Fisher's Exact test) ( 2007b).The test substance ( tested in solution) was negative in the Unscheduled DNA Synthesis (UDS) test in mammalian cells in vivo, following OECD Guideline 486; EC Annex V to Directive 67/548/EEC, Directive 2000/32/EC, B.39; OPPTS Guideline 870.5550; ICH S2A document April 24, 1996; and ICH S2B document November 21, 1997.

For the pilot toxicity assay, the test substance was administered via oral gavage to male rats at 10, 100, 1000, and 2000 mg/kg body weight in a total volume of 10 mL/kg bw. All animals treated with <1000 mg/kg appeared normal less than 4 hours following dosing, and 1, 2, and 3 days following dosing. All animals treated with 2000 mg/kg appeared normal less than 4 hours following dosing. However, on Day 1 following administration, they were lethargic, with 3 animals having crusty noses and 1 animal having crusty eyes. On Day 2, one animals was found dead, while the remaining appeared normal on Days 2 and 3. Based on the information from the pilot toxicity assay, the high dose for the UDS assay was set at the maximum tolerated dose, which was 2000 mg/kg, accompanied by two lower doses of 500 and 1000 mg/kg.

In the UDS test, the test substance was administered to 5 male rats per dose for 2 to 4 hours and 12 to 16 hours at doses of 500 and 1000 mg/kg. The test
substance was administered to 7 male rats for 2 to 4 hours and 12 to 16 hours at 2000 mg/kg to ensure that at least three animals would be available for UDS evaluation. Two additional groups of 5 rats each, for each time point, received a single oral dose of distilled water or 35 mg/kg dimethylnitrosamine (DMN), which served as the vehicle and positive control, respectively. The vehicle control, positive control, and test substance were administered at a constant volume of 10 mL/kg bw by a single oral gavage injection. No mortality or clinical signs were observed in any of the test substance or control

- treated harvested animals immediately following dosing or prior to harvest.

The group mean net nuclear grain (NG) counts for animals treated with the test substance were not increased when compared to the vehicle control. Forthe 2 to 4 hour time point, the group mean NG counts for the test substancetreated animals were -5.1, -6.5, and -7.2 with 1% of cells in repair (cells with >5 NG) for the 500, 1000, and 2000 mg/kg animals, respectively. The group mean NG count for the vehicle control was -5.8 with 0% of cells in repair. For the 12- to 16-hour time point, the group mean NG counts for the test substance-treated animals were -4.5, 4.6, and -4.2 with <3% of cells in repair (cells with >5 NG) for the 500, 1000, and 2000 mg/kg animals, respectively. The group mean NG count for the vehicle control was -7.1 with 1% of cells in repair. The positive control group mean NG counts were 14.3 and 13.8 for the 2 to 4 and the 12 to 16 hour, respectively. The percentage of cells in repair for the positive control group was 92% and 93% for the 2 to 4 hour and 12 to 16 hour, respectively. The test substance did not induce a significant increase in the mean number of net nuclear grain counts (i.e., an increase of at least 5 counts over the vehicle control group) in hepatocytes isolated either 2 to 4 hours or 12 to 16 hours after dose administration ( 2007c). In Vivo Gentic Toxicity Mammalian ToxicityIn Vivo Gentic Toxicity Mammalian Toxicity The test substance ( tested in solution) was negative for clastogenicity in the micronucleus and chromosome aberration assays, following OECD Guidelines 474 and 475; EC Commission Directive 2000/32/EC Annex 4B B11 and 4C-B12 No. L 136; and US EPA OPPTS Guideline 870.5395 and 870.5385.

The study was performed in two phases. The first phase, the pilot toxicity study, was designed to assess the toxicity of the test substance and to set dose levels for the definitive phase of the study. The second phase, the definitive study, was designed to evaluate the potential of the test substance to increase the incidence of chromosome aberrations in bone marrow cells (chromosome aberration assay) and the incidence of micronucleated polychromatic erythrocytes in bone marrow (micronucleus assay) of male and female ICR mice. Sterile water was used as the test substance vehicle. In the definitive phase of the study, water was used as the vehicle (negative) control substance and cyclophosphamide monohydrate (CP), at a dose of 50 mg/kg, as the positive control substance. In both phases of the study, test or control substances were administered as a single oral gavage.

In the pilot toxicity study, two male mice each were exposed to the test substance at 1, 10, 100, or 1000 mg/kg while five male and five female mice were exposed at 2000 mg/kg. No mortality was observed. Piloerection was seen in all males at doses >10 mg/kg and all females at 2000 mg/kg. Also at 2000 mg/kg lethargy (males and females) and hunched position (females) were observed. All males at 1 mg/kg appeared normal following dose administration and throughout the observation period. Reductions in the mean group animal body weights were observed in some of the groups during the course of the study. In the absence of mortality in the pilot toxicity study, the high dose for the definitive study was set at 2000 mg/kg.

The definitive phase of the study consisted of four treatment groups, each containing 10 male and 10 female mice with the exception of the high dose group that had total of 15 animals/sex. Doses were 0, 500, 1000, or 2000 mg/kg. Five male and five female mice per each treatment group were euthanized 24-hour post-dose and the remaining five animals per sex and treatment were euthanized at 48 hour post-dose. An additional 5 male and 5 female mice were dosed with the positive control substance (CP) and were euthanized 24-hour post-dose. From each animal, at the time of euthanasia, bone marrow from one femur was collected and processed for micronucleus analysis and the bone marrow from the other femur was processed for analysis of chromosome aberrations.

Bone marrow cells [polychromatic erythrocytes (2000 PCEs/animal)] from all 24 hour groups and the vehicle and the high dose 48-hour groups were examined microscopically for the presence of micronuclei (MPCEs). One thousand total erythrocytes were evaluated for cytotoxicity. Numerical and structural chromosome aberrations were analyzed in 100 metaphase cells per animal from all 24-hour groups and the vehicle and high dose 48-hour groups. In addition, the mitotic index was recorded as the percentage of cells in mitosis based upon a total of 1000 cells per animal.

No mortality was observed during the definitive study. Piloerection was seen in 3/15 males at 2000 mg/kg. All other males at 2000 mg/kg and all males and females treated with the lower doses or the control substances appeared normal following dose administration and throughout the course of the study. No statistically significant or biologically relevant reductions in the PCEs/ECs ratio or incidence of structural or numerical chromosome aberrations in test substance-treated groups relative to the respective vehicle control groups in male or female mice at 24 or 48 hours after dose administration was observed. Dose-dependant reductions in the mitotic index relative to the vehicle control groups of up to 54.2% and 51.7% in the male and female 24-hour test substance-treated groups, respectively, were observed. At 48-hour post-dose, a reduction in the mitotic index of 25.4% was observed in the male test substance-treated group. The observed reductions in mitotic index suggest that the test substance reached the target cells. CP, the positive control, induced a statistically significant increase in the incidence of micronucleated PCEs and a statistically significant increase in the incidence of
structural chromosome aberrations in both male and female mice. The incidence of micronucleated PCEs and chromosome aberrations in the vehicle control groups did not exceed the historical vehicle control range. Based upon this, all criteria for a valid test were met as specified in the protocol.

In conclusion, under the conditions described in this report, a single oral administration of the test substance at doses up to and including 2000 mg/kg, did not induce a significant increase in the incidence of micronucleated polychromatic erythrocytes or structural or numerical chromosome aberrations in bone marrow of male or female ICR mice. This indicates that the test substance was negative for
induction of chromosome aberrations in bone marrow cells and micronucleated polychromatic erythrocytes in bone marrow and therefore was concluded to have no genotoxic/clastogenic potential. The test substance was concluded to be negative in this in vivo study ( 2007d).Mammalian Toxicity Metabolism
Studies Male and female rats were given single oral doses (10 or 30 mg/kg) the test
substance ; tested in solution) in a biopersistence and pharmacokinetic screen. Three rats per sex per dose level were used. Blood was collected at 0, 0.25, 0.5, 1, 2, 4, 8, 12, 24, 48, 72, 96, 120, 144, and 168 hours. Fat and liver were analyzed for parent compound to provide an estimate of tissue:plasma ratio.

At sacrifice, fat:plasma ratio in females was below the limit of quantification (LOQ). The ratios in males given 10 or 30 mg/kg were <0.1 and 0.1, respectively. At sacrifice, liver:plasma ratios in females were calculated since plasma values were below the LOQ. The ratios in males given 10 or 30 mg/kg were 3 and 3.1, respectively ( 2007j).

Mammalian Toxicity Metabolism
Studies The rate and extent of urinary elimination of the test substance tested in solution) found in male rat urine 96 hours after dosing was studied. Three male rats were given a single oral dose of 73 μM/kg. Urine was sampled at 6, 24, 48, 72, and 96 hours. Recovery was 68% by LC/MS and 65% by NMR. The urine t1/2 was 23.7 and 20.9 hours determined by LC/MS and NMR, respectively ( 2007k).

Mammalian Toxicity Metabolism
Studies Live and heat inactivated male rat liver S9 (2 mg/mL protein concentration) fractions were exposed to 2.5uM ( for a total of two hours. At the end of two hours, the reaction was quenched by the addition of acetonitrile (1:2, sample:acetonitrile) and analyzed for parent compound by
LC/MS. The final concentration of was 129.1 (+/- 22.6) and 106.7 (+/- 6.0) mg/mL from the live and heat inactivated control samples respectively. was not metabolized by male rat liver S9 under the conditions of this assay ( 2009a).

Ecotoxicology Environmental

Fate Biodegradation The ready biodegradability was determined with a non-adapted activated sludge for over a test period of 29 days in the Modified Sturm Test. was tested at a concentration of 150 mg/L in duplicates, corresponding to a carbon content (TOC) of 10.1 mgC/L in the test vessels. The biodegradation of was followed by titrimetric analyses of the quantity of CO2 produced by the respiration of bacteria. The degradation was finished on day 28 by acidification, the last titration was made on day 29, after the soluble CO2 was turned out over a period of 24 hours. The percentage CO2 production was calculated in relation to the theoretical CO2 (ThCO2) of The biodegradation was calculated for each titration time. To check the activity of the test system, sodium acetate was used as the functional control. For sodium acetate, the pass level of 60% was reached after 8 days therefore showing the validity of the test system.

For the 10% level (beginning of biodegradation) was reached after 13 days. The pass level of 60% was not reached, neither in the 10-day window nor after 28 days. The mean biodegradation of came to a maximum of 42% after 28 days and therefore, should be regarded as not readily biodegradable ( 2006).Ecotoxicology Environmental

Fate Biodegradation Not Applicable Environ. Sci.

Technol ., 32:2283-
2287. Partial defluorination was observed in an isolate of Pseudomonas sp. strain D2 under aerobic, sulfur limiting conditions in mineral medium in a laboratory study (Key et al., 1998).

Key BD, Howell RD, and Criddle CS (1998). Defluorination of organofluorine sulfur compounds by Pseudomonas sp. strain D2. Environ. Sci. Technol.,
32:2283-2287.

Ecotoxicology Aquatic
Studies Fish 96-hour LC50 (rainbow trout, Oncorhynchus mykiss) >108 mg/L.

The acute toxicity of the test substance ( tested in solution) to rainbow trout was determined in an unaerated, static acute 96- hour limit test following OECD Guideline 203. Nominal concentrations tested were 0 and 120 mg/L. The mean water temperature was 12.4°C (range of 12.3-12.5°C). One test chamber was used per test concentration with 10 fish in each chamber. Control fish ranged from 3.3 to 4.3 cm in standard length (mean 3.93 cm) and 0.380 to 0.938 g in wet weight, blotted dry (mean 0.707 g) at test end. Control loading at test end was 0.354 g/L.

Mean measured concentrations were none detected (limit of detection 0.4 μg/L) and 108 mg/L for the 0 and 120 mg/L concentrations, respectively. No mortality or sublethal effects were observed at any concentration. Dilution water quality was acceptable based on OECD criteria and specifications. All chemical and physical parameters were within the expected ranges. Total alkalinity, EDTA hardness, and conductivity of the dilution water control and highest test substance concentration at test start ranged from 47 to 48 mg/L as CaCO3, 102 to 121 mg/L as CaCO3, and 245 to 255 μmhos/cm, respectively. During the test, dissolved oxygen concentrations ranged from 7.6 to 9.9 mg/L, pH ranged from 7.0 to 7.2, and mean temperature was 12.4°C with a range of 12.3 to 12.5°C ( 2008b).

Ecotoxicology Aquatic
Studies Invertebrates 48-hour EC50 (Daphnia magna) > 112 mg/L.

The acute toxicity of the test substance ( tested in solution) to unfed Daphnia magna neonates, less than 24 hours old at test start was determined in an unaerated, static, acute, 48 hour limit test following OECD Guideline 202. Nominal concentrations tested were 0 and 120 mg/L. The mean water temperature was 19.8°C (range of 19.6-19.9°C). Four replicates with five daphnids per replicate were used per test substance
concentration and control.

Mean measured concentrations were none detected (limit of detection 0.7 μg/L) and 112 mg/L for the 0 and 120 mg/L concentrations, respectively. No immobility or sublethal effects were observed at any concentration at the end of the 48-hour limit test. Dilution water quality was acceptable based on OECD criteria and specifications. All chemical and physical parameters were within the expected ranges. Total alkalinity, EDTA hardness, and conductivity of the dilution water control and highest test substance concentration at test start ranged from 49 to 50 mg/L as CaCO3, 121 to 123 mg/L as CaCO3, and 245 to 250 μmhos/cm, respectively. During the test, dissolved oxygen concentrations ranged from 8.5 to 9.2 mg/L, pH ranged from 7.2 to 8.0, and mean temperature was 19.8°C with a range of 19.6 to 19.9°C ( 2008c).Ecotoxicology Terrestrial
Studies Soil Dwelling Organisms 6:2 fluorotelomer sulfonate (6:2 FTS) Not Applicable BCF were determined for both adults and offspring and was 3.0 and 2.7, respectively (SFT, 2006).

Statens forurensningstilsyn (Norwegian Pollution Control Authority) [SFT] (2006). Økotoksikologiske effekter av PFOS, PFOA og 6:2 FTS på meitemark (Eisenia fetida), TA-2212/2006 (December) (http://www.sft.no/publikasjoner/kjemikalier/2212/ta2212.pdf ; accessed January 7 2009).

Ecotoxicology Terrestrial
Studies Soil Dwelling Organisms 6:2 fluorotelomer sulfonate (6:2 FTS) Not Applicable Exposing earthworms to soils from the four fire training facilities resulted in mean bioaccumulation factors (BAFs) for 6:2 fluoroteleomer sulphonate (6:2 FTS), PFOS and PFOA of 2.4 (0.76-6.7), 2.6 (0.49-6.4) and 5.9 (0.31-14.0), respectively (SFT, 2008).

Statens forurensningstilsyn (Norwegian Pollution Control Authority) [SFT] (2008). Screening of polyfluorinated organic compounds at four fire training facilities in Norway (http://www.klif.no/publikasjoner/2444/ta2444.pdf).

Ecotoxicology Terrestrial
Studies Soil Dwelling Organisms Not Applicable Accumulation experiments using standard soil (OECD-soil) have shown that earthworms bioaccumulate PFOS, PFOA and 6:2 FTS encountering BAFs of about 1-3. 6:2 FTS had somewhat higher BAFs than PFOS and PFOA (Stubberud, 2006).

Stubberud, H. 2006. Økotoksikologiske effecter av PFOS, PFOA og 6:2 FTS på meitemark (Eisenia fetida). (in Norwegian, English summary). Report TA 2212/2006. Norwegian State Pollution Control Authority, PB 8100 Dep, 0032 Oslo (cited in SFT, 2008). 72-hour EbC50 (green alga, Pseudokirchneriella subcapitata) > 125 mg/L (cell count). 0-72-hour EbC50 (green alga, Pseudokirchneriella subcapitata) > 125 mg/L (area under the curve). 72-hour ErC50 (green alga, Pseudokirchneriella subcapitata) > 125 mg/L (growth rate).

The toxicity of the test substance ( tested in solution) to the green algae was determined in a 72 hour, static limit test following OECD Guideline 201. The study was conducted with a blank control, an abiotic control, and 1 nominal concentration of the test substance (125 mg/L) at a mean lighting intensity of 7250 lux (range of 6760 to 7570 lux), a mean temperature of 24.0°C (range of 24.0-24.1°C), and a shaking speed of 104 rpm. Synthetic algal-assay-procedure (AAP) nutrient medium was used as the
test diluent and blank control. Test solutions were not renewed. Six replicates were used for the limit test concentration and blank control. A single test flask was used for the abiotic (stability) control. Healthy cell count, area under the growth curve, and growth rate were determined at 24-hour
intervals over the 72-hour test.

Mean measured concentrations were none detected (limit of detection 0.2 μg/L), 125, and 121 mg/L for the blank control, 120 mg/L, and abiotic control (120 mg/L) concentrations, respectively. Inhibition of growth expressed as cell count (density), area under the growth curve (biomass), and average specific growth rate of Pseudokirchneriella subcapitata exposed to a mean, measured limit test concentration of 125 mg/L for 72 hours was 20, 22, and 4%, respectively. Healthy cell counts increased in the blank control by at least a factor of 16 in 72 hours and the coefficient of variation of average specific growth rates during the whole test period (0-72 hour) in replicate controlcultures did not exceed 7%, and the mean coefficient of variation for sectionby-section specific growth rates (days 0-1, 1-2, and 2-30 in the blank control replicates did not exceed 35%, thereby satisfying the appropriate test acceptance criteria ( 2008d). Ecotoxicology Aquatic
Studies Other