### SAT Report PMN Number: P-13-0674 SAT Date: 7/16/2013 Print Date: 5/5/2014

Related cases : Health related cases: Ecotox related cases : N o suitable analogs were found.

Concern levels :
Type of Concern: Health Eco Comments Level of Concern: 1-2 2

Persistence Bioaccum Toxicity Comments 1 1 2 PMN 1 1 Hyd Pdt

Exposure Based Review : Health: N o Exposure based testing (health): XB testing through developmental toxicity testing submitted with PMN Ecotox: N o

Routes of exposure : Health: Drinking Water Inhalation Ecotox: All releases to water Fate: ; Hyd Pdt moderate

P 2 Rec Comments : Comment: P2REC: SAT: Do Not Forward. This substance showed developmental toxicity and was judged by SAT to have a potential to cause cancer

Keywords : Keywords: DEVEL; SYSTEMIC; ONCO (UNCERT); AQUATOX

Summary of Assessment :

Fate : Fate Summary: P-13-0674 FATE: Estimations for hydrolysis product log Kow = 0.35 (E) log Koc = 2.21 (E)log Fish BCF = 0.50 (E) log Fish BAF = 0.04 (E) PMN Substance: Solid with MP = Dec. 156.9 C (M) log Kow = -3.05 at pH 5 (M) S = 424 g/L at pH 4 and 25 °C (M) S = Reacts at pH 7 and 9 (M) Hydrolysis Half-life = min-hr at pH 7 or 9, >mo at pH 4 VP < 1.0E-6 torr at 25 C (E) BP > 400 C (E) H < 1.00E-8 (E) POTW removal (%) = PMN 90-99 via hydrolysis; then Hyd Pdt 90 via sorption and biodeg; OECD 302C(MITI): 100%/27d; OECD NS(Closed Btl): 63.7-64.1%/28d; OECD 301F(Mano Resp): 100%/28d; OECD 111(Hydrolysis): t1/2( p H4,7,9):276d, 3.5hr, 10min. Time for complete ultimate aerobic biodeg = Hyd Pdt wk Sorption to soils/sediments = Hyd Pdt moderate PBT Potential: PMN P1B1; Hyd Pdt P1B1 *CEB FATE: Migration to ground water = Hyd Pdt moderate

Health : Health Summary: N ot absorbed through the skin, moderate absorption from the GI tract and good absorption from the lung ( p che m ). Concern for developmental toxicity and general systemic toxicity based on submitted test data. Uncertain concern for oncogenicity based on the possible mechanism of action for the indicated use. Test Data: (-) Salmonella with and without activation; (-) for chromosome aberrations in V79 cells with and without activation; (-) for gene mutations in V79 cells with and without activation; (-) ip mouse micronucleus assay; rat oral LD0 = 2000 mg/kg; rat dermal LD0 = 2000 mg/kg; rat inhalation LC0 = 5.75 mg/L (5750 mg/m3); no skin irritation in rabbits; not corrosive to human skin in vitro at 85 and 32% ai; not irritating to human skin in vitro at 85 and 32% ai; no eye irritation in rabbits; no serious eye damage in vitro Bovine Corneal Opacity and Permeability Test; (-) for skin sensitization in a mouse local lymph node assay up to 50% ai; rat oral developmental toxicity NOEL = 250 mg/kg; rat 28-day oral NOEL = 1000 mg/kg (highest dose tested); rat 90-day oral NOEL = 300 mg/kg, lower mean body weight and elevated bile acids in blood at 1000 mg/kg

Ecotox :

Test Organism Test Type Test End Point Predicted Measured Comments

fish 96-h LC50 >100 60.0 daphnid 48-h LC50 >100 24.7 green algal 96-h EC50 >100 12.9 48-hour test results; yield fish

_ chronic value >10 daphnid

_ chronic value >10algal

_ chronic value >10 0.106 48-hour test results; yield Sewage Sludge 3-h EC50

_ Sewage Sludge

_ Chronic Value _

Ecotox Values Comments: Predictions are based on SARs for anionic surfactants: SAR chemical class = surfactant-anionic ; MW 376; solid with mp = 150 C (dec., M); S = 424 g/L at 20 C (M); p H7; effective concentrations based on 100% active ingredients and nominal concentrations; hardness < 150.0 mg/L as CaCO3; and TOC <2.0 mg/L;

Ecotoxicity Test Data Results for P-13-0674:

Acute fish, daphnia, and algae toxicity studies were completed for PMN P-13-0674 in 2009 by A second acute fish study was completed for PMN P-13-0674 by but was submitted as a robust summary without full study details. The PMN material is described as a solid in the neat form, as a solid mixture

The molecular weight is 376.32 g/mol. Based on an experimental water solubility of 424 g/L, an estimated Log Kow of 0.35 and an estimated Henry’s Law Constant of < 1.00 x 10-8 atm-m 3/mole this PMN is expected to nonvolatile and highly soluble in water. The PMN material also reacts at pH 7 and 9 based on experimental results and has experimental hydrolysis half lives at 25 °C and pH 4, p H 7 and pH 9 of 276 days, 3.5 hours and 10 minutes, respectively. The PMN material is The pH is controlled at > 8

– 9 during processing. The PMN material is used as

Studies followed the respective guidelines of OECD 203 (Acute Toxicity for Fish), Council Regulation (EC) N o 440/2008, Method C.1 'Acute toxicity for Fish' (2008), OECD 202 (Daphnia sp., Acute Immobilization Test), Council Regulation (EC) N o 440/2008, Method C.2 ‘Acute toxicity for Daphnia’ (2008), OECD 201 (Freshwater Alga Growth Inhibition Test) and Commission Regulation (EC) N o 761/2009 amending Regulation No 440/2008, Method C.3 ‘Freshwater Algae and Cyanobacteria, Growth inhibition test’ (2009) and were GLP compliant. Study methods concurred with methods specified in the respective guidelines except where specified below.

Fish Ecotoxicity Test: (1) conducted a 96-hour acute toxicity test in zebra fish (Brachydanio rerio) with P-13-0674 under semi-static conditions with 24-hour renewal. The substance tested was a solution containing 65% water and 35% solid. The solid is the “drying residue” and contains 31.95% of the PMN substance and is considered to be the active ingredient. Following a range-finding test, single replicates of seven B. rerio were exposed to adilution water control (reconstituted water) or the test substance at nominal test concentrations of 9.4, 20.7, 45.5 and 100 mg/L. Effective concentrations ranged from 97.9

– 111.4% of nominal values in the freshly prepared media and from 44.1

– 82.8% of nominal values in the media after 24 hours of exposure. Concentrations were only measured for the 20.7 and 100 mg/L nominal test concentrations. The test item concentrations for the 9.4 and 45.5 mg/L test concentrations were calculated based on the mean recovery rate measured at the next highest concentration, respectively. Thus, the recovery rate for the 20.7 mg/L nominal test concentration was used to calculate the concentration of the 9.4 mg/L nominal test concentration. Measured concentrations were determined by HPLC analysis and MS detection with a limit of quantification (LOQ) of 0.021 mg/L. The analytical values refer to the drying residue of the test substance with a content of 35.05%. Stock solutions were prepared to give the desired series of test concentrations based on a drying residue of 35.05% (according to data of the sponsor). To prepare a stock solution, a certain amount of test substance was added daily to 1 L of Millipore water and shortly treated on a magnetic stirrer. Appropriate amounts of the stock solution were then diluted with water according ISO 7346 and adjusted to a pH of 6 to obviate rapid hydrolysis. Test vessels were covered with glass plates. Over the course of testing, temperature ranged from 19.9

– 21.5 °C,
p H ranged fro m 6.0

– 6.7 and dissolved oxygen ranged from 7.6

– 9.0 mg/L. Dilution water hardness was 264 mg CaCO3/L which exceeds both the OECD 203 guideline water hardness value of 250 mg CaCO3/L and the OPPTS 1075 water hardness value of 180 mg CaCO3/L. A loading rate of 0.54 g fish/L was reported. Observations were made on the numbers of dead fish and the incidence of sub-lethal effects after 2, 24, 48, 72 and 96 hours of exposure. N o mortalities occurred at nominal concentrations ≤ 45.5 mg/L. However, there was abnormal swimming behavior documented after 2 hours at the nominal concentrations of 45.5 mg/L and 100 mg/L. Cumulative mortality in the 100 mg/L group was 86%. Based on geometric mean measured concentrations, the 96-hour LC50 is 65.86 mg/L for the drying residue and 60.01 mg/L for the active ingredient. Although the water hardness exceeded the test guideline values, this test was considered to be adequate. 96-hour LC50 = 65.86 mg/L (drying residue) 96-hour LC50 = 60.01 mg/L (active ingredient)

(2) conducted a 96-hour acute toxicity test in zebra fish (Brachydanio rerio) with P-13-0674 (98.2 ± 1.2% p urity) under semi-static conditions with 24-hour renewal. The test temperature was 23 ± 1°C. Only a study summary is available; no additional study details were provided. The 96-hour LC50 is 67.2 mg/L (it is not specified whether this value is based on nominal or measured concentrations). This test is not considered to be adequate due to the lack of information on the test conditions and results. 96-hour LC50 = 67.2 mg/L

Invertebrate Ecotoxicity Test: conducted a 48-hour acute toxicity test in water fleas (Daphnia magna) with P-13-0674 under semi-static conditions with 24-hour renewal. The substance tested was a solution containing 65% water and 35% solid. The solid is the “drying residue” and contains 31.95% of the PMN substance and is considered to be the active ingredient. Following a range-finding study, two replicates of 10 D. magna were exposed to a dilution water control or the test substance at nominal concentrations of 1.9, 4.3, 9.4, 20.7, 45.5 and 100 mg/Lcorresponding to geometric mean concentrations of 0.922, 1.61, 4.49, 10.9, 27.1 and 63.3 mg/L, respectively. Recovery rates ranged from 83.0

– 98.7% of nominal values in freshly prepared media, and from 14.8

– 56.5% of nominal values in media after 24 hours of exposure, respectively. Measured concentrations were determined by HPLC analysis and MSD-detection with a limit of quantification (LOQ) of 0.021 mg/L. The analytical values refer to the drying residue of the test substance with a content of 35.05%. Stock solutions were prepared to give the desired series of test concentrations based on a drying residue of 35.05% (according to data of the sponsor). To prepare a stock solution, a certain amount of test substance was added to 1 L of distilled (Millipore) water. Appropriate amounts of the stock solution were then diluted with M4 Medium which was adjusted to a pH of 6 to obviate rapid hydrolysis of the test item. Test vessels were covered with glass plates. Over the course of testing, temperature ranged from 19.5

–

21.9 °C, p H ranged fro m 6.0

– 7.0 and dissolved oxygen ranged from 8.0

– 8.9 mg/L. Dilution water hardness was 266 mg CaCO3/L which exceeds both the OECD 203 guideline water hardness value of 250 mg CaCO3/L and the OPPTS 1010 water hardness value of 180 mg CaCO3/L. A loading rate of 500 daphnia/L was calculated exceeding the loading rate of 40 daphnids/L in the OPPTS guideline, p otentially causing overcrowding. N o immobilization occurred at concentrations ≤ 10.9 mg/L. Percent immobilization at 27.1 and 63.3 mg/L was 50% and 100%, respectively. Additionally, abnormal swimming behavior was observed at 63.3 mg/L. Based on geometric mean measured concentrations, the 48-hour EC50 is 27.1 mg/L for the drying residue and 24.7 mg/L for the active ingredient. Although the water hardness exceeded the test guideline values and the daphnid loading exceeded the OPPTS guideline, this test was considered to be adequate. 48-hour EC50 = 27.1 mg/L (drying residue) 48-hour EC50 = 24.7 mg/L (active ingredient)

Algal Ecotoxicity Test: conducted a 72-hour toxicity test in green algae (Desmodesmus subspicatus) with P-13-0674 under static conditions. The substance tested was a solution containing 65% water and 35% solid. The solid is the “drying residue” and contains 31.95% of the PMN substance and is considered to be the active ingredient. While the study was conducted for 72 hours, the data were analyzed on a 48 hour basis since the algal cell numbers increased well above 16-fold over 48 hours and all other validity criteria were met. This option is cited to be covered by OECD 201. The OPPTS guideline requires a 96-hour algal test. The substance tested in this study was not fully characterized and its identity was not provided. The active ingredient was described and confirmed to be the PMN substance. Following a range-finding test, three replicates of D. subspicatus (5000 cells/mL) were exposed to a dilution water control or the test substance at nominal concentrations of 0.32, 1.0, 3.2, 10 and 32 mg/L. Corresponding geometric mean measured concentrations of 0.071, 0.189, 0.467, 1.755 and 8.413 mg/L were determined via HPLC analysis and MSD-detection with a limit of quantification (LOQ) of 0.021 mg/L. The analytical values refer to the drying residue of the test substance with a content of 35.05%. Effective concentrations ranged from 76.0

– 103.4% of nominal values at 0 hours, from 16.2

– 35.1% of nominal values at 24 hours, and from 2.5

– 9.6% of nominal values at 48 hours, respectively. Stock solutions were prepared to give the desired series of test concentrations based on a drying residue of 35.05% (according to data of the sponsor). To prepare a stock solution, 356.8 mg of test substance was added to 1 L of dilutionwater and adjusted to a pH of 6 to obviate rapid hydrolysis of the test item. To produce the different test concentrations, the prepared stock solution was diluted with dilution water to a volume of 80 mL. This was followed by the addition of 10 mL of nutrient medium and 10 mL of algal inoculum with a cell density of 5000 cells/mL to achieve the final nominal concentrations. Algal cultures were illuminated at a light intensity of 4000

– 8000 lux with constant shaking. Over the course of testing, temperature ranged from 20.0

– 21.5 °C and pH ranged from 6.1

– 7.2. Dilution water hardness was 22.5 mg CaCO3/L. Mean cell density in control cultures increased by a factor of 108.7 within 72 hours. The test was carried out for 72 hours, but only 48-hour results are reported. Based on geometric mean measured concentrations, the 48-hour EC50 values for growth rate and yield are > 100 and 12.9 mg/L, respectively, for the active ingredient. The NOEC and LOEC values for the active ingredient are 0.065 and 0.172 mg/L, respectively. The ChV for the active ingredient is 0.106 mg/L. Because concentrations of the test item markedly decreased over time, the study data were analyzed on a 48-hour basis which reflects exposure better than the 72-hour values based on comments from the testing facility. Minimally, the results for the 72-hour test duration should have been reported, but the 48-hour results are considered to be adequate since algae is the most sensitive species using the 48-hour results. Drying Residue (35.05%): 48-hour ErC50 (growth rate) > 100 mg/L 48-hour EyC50 (yield) = 14.1 mg/L 48-hour NOEC (growth rate and yield) = 0.071 mg/L 48-hour LOEC (growth rate and yield) = 0.189 mg/L

Active Ingredient (31.94%): 48-hour ErC50 (growth rate) > 100 mg/L 48-hour EyC50 (yield) = 12.9 mg/L 48-hour NOEC (growth rate and yield) = 0.065 mg/L 48-hour LOEC (growth rate and yield) = 0.172 mg/L

The 96-hour fish (LC50 = 60.01 mg/L), 48-hour daphnid (EC50 = 24.7 mg/L) and 48-hour algal (EyC50 = 12.9 mg/L; N OEC = 0.065 mg/L and LOEC = 0.172 mg/L) toxicity tests were considered to be adequate for P-13-0674 (the active ingredient). For comparative purposes, the 96-hour fish LC50, the 48-hour daphnid LC50, and the 96-hour algae EC50 of >100 mg/L, and the fish, daphnid, and algae chronic values (ChV) of > 10 mg/L, respectively were predicted using the SAR for anionic surfactants. Available experimental data indicate algae to be the most sensitive species for calculating the acute and chronic concentrations of concern (CoC). The 48-hour algae results are being used to calculate the acute and chronic CoCs, however, if risk is confirmed from environmental releases based on the CoC values below, EPA recommends the submittal of the results for the full 72-hour test or for the algae test to be conducted again for the full 96-hours with the PMN compound and hydrolysis product measured over the duration of the test. Based on the experimental 48-hour algae EyC50 of 12.9 mg/L (12,900 p pb), and the application of an assessment factor of 4, the acute CoC for P-13-0674 is 3225 p pb. The ChV for P-13-0674 is calculated by determining the geometric mean of the NOEC and LOEC to yield a ChV of 0.106 mg/L (106 p pb). The chronic CoC for P-13-0674 is then derived by dividing the ChV of 0.106 mg/L (106 p pb) by a factor of 10 (uncertainty factor) yielding a chronic CoC of10.6 p pb.

Acute CoC = 3225 p pb Chronic CoC = 11 p pb

Ecotox Study Reviewer: K. Eisenreich July 24, 2013

Factors Values Comments Assessment Factor 10 Concentration of Concern ( p pb) 11

SARs anionic surfactants SAR Class surfactant-anionic Ecotox Category Anionic Surfactants

Ecotox Factors Comments:

SAT Chair: L Keifer 564-8916