oEPA

United States Environmental	Office of Water	EPA-822-R-24-009

Protection Agency	4304T	September 2024

Acute Freshwater Aquatic Life Benchmarks

for Eight Data-Limited PFAS:

PFBA, PFHxA, PFNA, PFDA, PFBS, PFHxS,
8:2 FTUCA, and 7:3 FTCA

September 2024

U.S. Environmental Protection Agency Office of Water,

Office of Science and Technology, Health and Ecological Criteria Division,

Ecological Risk Assessment Branch
Washington, D.C.


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Table of Contents

Table of Contents	ii

List of Appendices	iii

List of Tables	iii

List of Figures	v

Project Team	vii

Acronyms	viii

Notices	ix

Foreword	xi

Executive Summary	xii

1	Background	1

2	Introduction to ICE	5

3	Application of ICE with Selected PFAS	8

4	Effects Analysis for Aquatic Life	11

4.1	Summary and Evaluation of Available Empirical Toxicity Test Data	11

4.2	Prediction of Species Sensitivity Using ICE Models	15

5	Derivation of Acute Water Quality Benchmarks for Selected PFAS	17

5.1	Approach	17

5.1.1	Use of Empirical and ICE Data	17

5.1.2	Application Factor to Adjust for Additional Unrepresented Sensitive Species

Data	19

5.2	Derivation of Acute Water Benchmark for Perfluorobutanoic Acid (PFBA)	21

5.3	Derivation of Acute Water Benchmark for Perfluorohexanoic Acid (PFHxA)	27

5.4	Derivation of Acute Water Benchmark for Perfluorononanoic Acid (PFNA)	33

5.5	Derivation of Acute Water Benchmark for Perfluorodecanoic Acid (PFDA)	39

5.6	Derivation of Acute Water Benchmark for Perfluorobutanesulfonic Acid (PFBS)	48

5.7	Derivation of Acute Water Benchmark for Perfluorohexanesulfonic Acid (PFHxS).. 57

5.8	Derivation of Acute Water Benchmark for Hexadecafluoro-2-decenoic Acid

(8:2 I I I ( A)	63

5.9	Derivation of Acute Water Benchmark for Pentadecafluorodecanoic Acid

(7:3 FTC A)	74

5.10	Summary of Benchmarks for Evaluated PFAS Substances and Effects
Characterization	85

5.10.1	Summary and Evaluation of Benchmark Values	85

5.10.2	Comparison of Benchmarks to Other Calculation Methods for Data-Limited
PFAS	91

5.10.3	Summary and Conclusions	95

6	References	99

Appendices	102

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List of Appendices

Appendix A Quantitatively Acceptable Freshwater Acute Toxicity Studies	A-l

Appendix B Summary of Lowest Quantitatively Acceptable Freshwater Acute Toxicity

Studies	B-l

Appendix C Qualitative Freshwater Acute Toxicity Studies	C-l

Appendix D Unused Freshwater Acute Toxicity Studies	D-l

Appendix E ICE Models Available in Web-ICE v4.0 for Freshwater Predicted Species

Based on Surrogates with Measured Toxicities	E-l

Appendix F Derivation of PFAS Benchmarks using Scaled Data	F-l

Appendix G PFAS Acute Benchmark Calculations using a Data Binning Approach	G-l

List of Tables

Table Ex-1. Acute Freshwater Aquatic Life Benchmarks for Eight PFAS	xv

Table 3-1. Measured Surrogate Species Sensitivity Values and Number of Available ICE

Models for Eight PFAS	8

Table 4-1. Summary Table of PFAS Forms and Associated CAS Numbers Identified for the

Evaluation of Available Empirical Data	11

Table 4-2. Number of Acute Genus and Species Level Mean Values in the Empirical Toxicity
Datasets and Freshwater Minimum Data Requirements Fulfilled per the Aquatic

Life Criteria Guidelines for the Eight PFAS	15

Table 5-1. Application Factors Applied to Adjust for Additional Unrepresented Sensitive

Species Data	21

Table 5-2. Acceptable Models for ICE-estimated Species Sensitivity to PFBA	23

Table 5-3. Ranked PFBA Genus Mean Acute Values	25

Table 5-4. PFBA Protective Aquatic Life Acute Benchmark	26

Table 5-5. Acceptable Models for ICE-estimated Species Sensitivity of PFHxA	29

Table 5-6. Ranked PFHxA Genus Mean Acute Values	31

Table 5-7. PFHxA Final Acute Value and Protective Aquatic Acute Benchmark	32

Table 5-8. Acceptable Models for ICE-estimated Species Sensitivity to PFNA	35

Table 5-9. Ranked PFNA Genus Mean Acute Values	37

Table 5-10. PFNA Final Acute Value and Protective Aquatic Acute Benchmark	38

Table 5-11. Acceptable Models for ICE-estimated Species Sensitivity to PFDA	41

Table 5-12. Ranked PFDA Genus Mean Acute Values	45

Table 5-13. PFDA Final Acute Value and Protective Aquatic Acute Benchmark	47

Table 5-14. Acceptable Models for ICE-estimated Species Sensitivity to PFBS	51

Table 5-15. Ranked PFBS Genus Mean Acute Values	55

Table 5-16. PFBS Protective Aquatic Life Acute Benchmark	56

Table 5-17. Acceptable Models for ICE-estimated Species Sensitivity to PFHxS	59

Table 5-18. Ranked PFHxS Genus Mean Acute Values	61

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Table 5-19. PFHxS Final Acute Value and Protective Aquatic Acute Benchmark	62

Table 5-20. Acceptable Models for ICE-estimated Species Sensitivity to 8:2 FTUCA	66

Table 5-21. Ranked 8:2 FTUCA Genus Mean Acute Values	71

Table 5-22. 8:2 FTUCA Final Acute Value and Protective Aquatic Acute Benchmark	73

Table 5-23. Acceptable Models for ICE-estimated Species Sensitivity to 7:3 FTCA	77

Table 5-24. Ranked 7:3 FTCA Genus Mean Acute Values	82

Table 5-25. 7:3 FTCA Final Acute Value and Protective Aquatic Acute Benchmark	84

Table 5-26. Acute Freshwater Benchmarks for Eight PFAS (mg/L, ppm)	87

Table 5-27. Summary of Data Inputs, MDRs Fulfilled and Benchmark Outcomes Using the
Extrapolation and Scaled Approach when Web-ICE Input PFAS Values Exceed

Model Range	89

Table 5-28. Comparison of PFAS Chain Length to Perfluorocarboxylic and Perfluorosulfonic

Acid Benchmarks and PFOA and PFOS Acute Criteria Values	90

Table 5-29. Comparison of Benchmark Outcomes with Acute Benchmarks Calculated Based
on Approach by Giddings et al. (2019) (Aquatic Life Criteria Guidelines and SSD

Generator-Based Values)	93

Table 5-30. Comparison of Benchmark Outcomes with Water Quality-Based Ecological
Screening Values Calculated by Grippo et al. (2021) Using the Great Lakes

Initiative (GLI) Approach	95

Table 5-31. Summary of the EPA Acute Benchmark Values and Acute Values Calculated

Using Alternative Methods	98

Table E-l. All ICE Models Selected for the Derivation of PFAS Benchmarks	E-l

Table F-l. Acceptable models for ICE-estimated Species Sensitivity to PFBA using the

scaling approach for data outside of the model bounds as in cases indicated	F-4

Table F-2. Ranked PFBA Genus Mean Acute Values	F-8

Table F-3. PFBA Protective Aquatic Life Acute Benchmark	F-9

Table F-4. Acceptable models for ICE-estimated Species Sensitivity to PFHxA using the

scaling approach for data outside of the model bounds as in cases indicated	F-12

Table F-5. Ranked PFHxA Genus Mean Acute Values	F-16

Table F-6. PFHxA Final Acute Value and Protective Aquatic Acute Benchmark	F-17

Table F-7. Acceptable models for ICE-estimated Species Sensitivity to PFNA using the

scaling approach for data outside of the model bounds as in cases indicated	F-20

Table F-8. Ranked PFNA Genus Mean Acute Values	F-24

Table F-9. PFNA Final Acute Value and Protective Aquatic Acute Benchmark	F-25

Table F-10. Acceptable models for ICE-estimated Species Sensitivity to PFDA using the

scaling approach for data outside of the model bounds as in cases indicated	F-28

Table F-l 1. Ranked PFDA Genus Mean Acute Values	F-34

Table F-12. PFDA Final Acute Value and Protective Aquatic Acute Benchmark	F-36

Table F-13. Acceptable models for ICE-estimated Species Sensitivity to PFBS using the

scaling approach for data outside of the model bounds as in cases indicated	F-39

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Table F-14. Ranked PFBS Genus Mean Acute Values	F-48

Table F-15. PFBS Final Acute Value and Protective Aquatic Acute Benchmark	F-50

Table F-16. Acceptable models for ICE-estimated Species Sensitivity to PFHxS using the

scaling approach for data outside of the model bounds as in cases indicated	F-53

Table F-17. Ranked PFHxS Genus Mean Acute Values	F-55

Table F-18. PFHxS Final Acute Value and Protective Aquatic Acute Benchmark	F-56

Table F-19. Acceptable models for ICE-estimated Species Sensitivity to 8:2 FTUCA using

the scaling approach for data outside of the model bounds as in cases indicated. ..F-59

Table F-20. Ranked 8:2 FTUCA Genus Mean Acute Values	F-65

Table F-21. 8:2 FTUCA Final Acute Value and Protective Aquatic Acute Benchmark	F-67

Table F-22. Acceptable models for ICE-estimated Species Sensitivity to 7:3 FTCA using

the scaling approach for data outside of the model bounds as in cases indicated. ..F-70

Table F-23. Ranked 7:3 FTCA Genus Mean Acute Values	F-76

Table F-24. 7:3 FTCA Final Acute Value and Protective Aquatic Acute Benchmark	F-78

Table F-25. Acute Freshwater Benchmarks for Eight PFAS	F-80

Table G-l. Acute LCsos, SMAVs, normalized Danio rerio equivalent values and rank of

SMAV D. rerio equivalent values for the sulfonic acid PFAS	G-5

Table G-2. Acute LCsos, SMAVs, normalized Daphnia magna equivalent values and rank

of SMAV D. magna equivalent values for carboxylic acid PFAS	G-l

Table G-3. Results of lognormal regression analysis of combined PFAS group SSDs based

on key species equivalents and using the EPA's SSD Generator	G-10

Table G-4. Calculated freshwater FAV based on four lowest SMAV key species

equivalents for sulfonic acid PFAS	G-10

Table G-5. Calculated freshwater FAV based on four lowest SMAV key species

equivalents for carboxylic acid PFAS	G-10

Table G-6. PFAS acute benchmarks for individual PFAS within sulfonic and carboxylic
acid groups calculated using approach presented by Giddings et al. (2019):

Summary of values determined from SSDs derived using Aquatic Life Criteria
Guidelines-based and SSD Generator-based values	G-l 1

List of Figures

Figure 2-1. Example of ICE Model for Rainbow Trout (surrogate) and Atlantic Salmon

(predicted)	7

Figure 3-1. Example Calculation for Daphnia magna PFBA Toxicity Data Based on

Extrapolation and Scaled Approaches	10

Figure 4-1. Chemical Structures of Selected PFAS	12

Figure 5-1. Ranked Acute PFBA GMAVs Used for the Aquatic Life Acute Benchmark

Calculation	27

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Figure 5-2. Ranked Acute PFHxA GMAVs Used for the Aquatic Life Acute Benchmark

Calculations	33

Figure 5-3. Ranked Acute PFNA GMAVs Used for the Aquatic Life Acute Benchmark

Calculation	39

Figure 5-4. Ranked Acute PFDA GMAVs Used for the Aquatic Life Acute Benchmark

Calculation	48

Figure 5-5. Ranked Acute PFBS GMAVs Used for the Aquatic Life Acute Benchmark

Calculation	57

Figure 5-6. Ranked Acute PFHxS GMAVs Used for the Aquatic Life Acute Benchmark

Calculation	63

Figure 5-7. Ranked Acute 8:2 FTUCA GMAVs Used for the Aquatic Life Acute Benchmark

Calculation	74

Figure 5-8. Ranked Acute 7:3 FTCA GMAVs Used for the Aquatic Life Acute Benchmark

Calculation	85

Figure F-l. Ranked Acute PFBA GMAVs Used for the Aquatic Life Acute Benchmark

Calculation	F-10

Figure F-2. Ranked Acute PFHxA GMAVs Used for the Aquatic Life Acute Benchmark

Calculations	F-l 8

Figure F-3. Ranked Acute PFNA GMAVs Used for the Aquatic Life Acute Benchmark

Calculation	F-26

Figure F-4. Ranked Acute PFDA GMAVs Used for the Aquatic Life Acute Benchmark

Calculation	F-37

Figure F-5. Ranked Acute PFBS GMAVs Used for the Aquatic Life Acute Benchmark

Calculation	F-51

Figure F-6. Ranked Acute PFHxS GMAVs Used for the Aquatic Life Acute Benchmark

Calculation	F-57

Figure F-7. Ranked Acute 8:2 FTUCA GMAVs Used for the Aquatic Life Acute

Benchmark Calculation	F-68

Figure F-8. Ranked Acute 7:3 FTCA GMAVs Used for the Aquatic Life Acute Benchmark

Calculation	F-79

Figure G-l. Species sensitivity distributions for sulfonic acid PFAS based on Danio rerio
equivalents, using the EPA's SSD Generator (left panel - A) or the Aquatic Life

Criteria Guidelines procedure (right panel - B)	G-l2

Figure G-2. Species sensitivity distributions for carboxylic acid PFAS based on Daphnia
magna equivalents, using the EPA's SSD Generator (left panel - A) or the
Aquatic Life Criteria Guidelines procedure (right panel - B)	G-l3

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Project Team

Technical Analysis Leads:

Mike Elias, Office of Water, Office of Science and Technology, Health and Ecological Criteria
Division, Ecological Risk Assessment Branch Division, Washington, DC (Lead)*

Kathryn Gallagher, Branch Chief, Office of Water, Office of Science and Technology, Health
and Ecological Criteria Division, Ecological Risk Assessment Branch Division, Washington, DC

Interspecies Correlation Estimate Model Contributors:

Crystal Lilavois, Office of Research and Development, Center for Environmental Measuring and
Modeling, Gulf Ecosystem Measurement and Modeling Division, Gulf Breeze, FL

Sandy Raimondo, Office of Research and Development, Center for Environmental Measuring
and Modeling, Gulf Ecosystem Measurement and Modeling Division, Gulf Breeze, FL

Reviewers:

Colleen Flaherty, Division Director, Office of Water, Office of Science and Technology, Health
and Ecological Criteria Division, Washington, DC

*Please contact Mike Elias at Elias.mike@epa.gov for any questions.

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Acronyms

AWQC

CAS

CWA

DER

ECOTOX

ECx

EPA

FAV

7:3 FTCA

8:2 FTUCA

GMAV

hpf

ICE

LCx

LOECs

MDRs

MSE

NAMS

NOECs

OCSPP

ORD

OW

PFAS

PFBA

PFBS

PFDA

PFHxA

PFHxS

PFNA

PFOA

PFOS

SMAV

SOP

SSD

U.S.

web-ICE
WWTPs

National Recommended Ambient Water Quality Criteria

Chemical Abstracts Service

Clean Water Act

Data Evaluation Record

ECOTOXicology database

Effect concentration at x percent level

U.S. Environmental Protection Agency

Final Acute Value

2H,2H,3H,3H-Pefluorodecanoic acid

2H-Perfluoro-2-decenoic acid

genus mean acute value

hours post fertilization

Interspecies Correlation Estimation

Lethal concentration at x percent level

Lowest Observed Effect Concentrations

minimum data requirements

Mean square error

New Approach Methods

No Observed Effect Concentrations

Office of Chemical Safety and Pollution Prevention

Office of Research and Development

Office of Water

per- and polyfluorinated substances
Perfluorobutanoic acid
Perfluorobutanesulfonic acid
Perfluorodecanoic acid
Perfluorohexanoic acid
Perfluorohexanesulfonic acid
Perfluorononanoic acid

Perfluorooctanoic acid, pentadecafluoro-l-octanoic acid,
pentadecafluoro-n-octanoic acid, octanoic acid, pentadecafluoro-,
perfluorocaprylic acid, pentadecafluorooctanoic acid, perfluoroalkyl
carboxylic acid or perfluoroheptanecarboxylic acid
Perfluorooctane sulfonic acid or perfluorooctane sulfonate
Species Mean Acute Value
standard operating procedure
species sensitivity distribution
United States

Web-based Interspecies Correlation Estimation
wastewater treatment plants

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This document provides information that states and authorized tribes may consider in
their water quality protection programs under the Clean Water Act (CWA) to protect freshwater
aquatic life from the acute effects of eight individual per- and polyfluoroalkyl substances (PFAS)
substances: Perfluorobutanoic acid (PFBA), Perfluorohexanoic acid (PFHxA),

Perfluorononanoic acid (PFNA), Perfluorodecanoic acid (PFDA), Perfluorobutanesulfonic acid
(PFBS), Perfluorohexanesulfonic acid (PFHxS), Hexadecafluoro-2-decenoic acid (8:2 FTUCA),
and Pentadecafluorodecanoic acid (7:3 FTCA). Toxicity data for these eight PFAS are limited
relative to recommended data requirements traditionally used to develop ambient water quality
criteria (AWQC) for the protection of aquatic life. Therefore, the United States Environmental
Protection Agency (EPA) derived acute PFAS aquatic life freshwater benchmark values under
Section 304(a)(2) of the CWA using the best available data, as informational values that states
and Tribes may consider in their water quality protection programs, including establishing their
water quality criteria. While this document contains the EPA's scientific analyses regarding
benchmark values for ambient freshwater concentrations of these eight PFAS protective of
aquatic life, this document does not substitute for the CWA or the EPA's regulations; nor is this
document or the benchmarks values for the PFAS it presents a regulation itself. Thus, this
document does not establish or affect legal rights or obligations, or impose legally binding
requirements on the EPA, states, Tribes, or the regulated community. It cannot be finally
determinative of the issues addressed. This document has been approved for publication by the
Office of Science and Technology, Office of Water, U.S. Environmental Protection Agency.


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Mention of trade names or commercial products does not constitute endorsement or
recommendation for use. This document can be downloaded from:
https://www.epa.gov/svstem/files/documents/2024-08/pfas-report-2024.pdf

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Foreword

This document presents acute freshwater benchmark values for aquatic life in ambient
water based upon consideration of all available toxicity information relating to the acute effects
of eight individual per- and polyfluoroalkyl substances (PFAS) substances on freshwater aquatic
organisms: Perfluorobutanoic acid (PFBA), Perfluorohexanoic acid (PFHxA), Perfluorononanoic
acid (PFNA), Perfluorodecanoic acid (PFDA), Perfluorobutanesulfonic acid (PFBS),
Perfluorohexanesulfonic acid (PFHxS), Hexadecafluoro-2-decenoic acid (8:2 FTUCA), and
Pentadecafluorodecanoic acid (7:3 FTCA). The EPA developed this document to provide
information that states and authorized Tribes may consider in their water quality protection
programs.

Deborah G. Nagle

Director, Office of Science and Technology

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Executive Summary

Per- and polyfluoroalkyl substances (PFASs) are organic, human-made chemical
compounds composed of an alkyl chain bound to multiple fluorine atoms. There are estimated to
be over 3,000 PFAS present in the global marketplace, that are used in a variety of specialized
applications, including surface coatings and a variety of other industrial and commercial
products (Wang et al. 2017). In addition to Perfluorooctanoic acid (PFOA) and Perfluorooctane
sulfonic acid (PFOS), for which the U.S. Environmental Protection Agency (EPA) has recently
developed Aquatic Life Ambient Water Quality Criteria (U.S. EPA 2024b,c), the EPA
recognizes that there may be thousands of other PFAS present in the environment. The EPA
(2024b,c) provides a detailed discussion of the sources, occurrence, fate and transport of PFOA,
PFOS and other PFAS in the environment.

The EPA has developed acute freshwater aquatic life acute ambient water quality
benchmarks for eight selected polyfluorinated substances. The eight chemicals for which acute
benchmarks were developed are: Perfluorobutanoic acid (PFBA), Perfluorohexanoic acid
(PFHxA), Perfluorononanoic acid (PFNA), Perfluorodecanoic acid (PFDA),
Perfluorobutanesulfonic acid (PFBS), Perfluorohexanesulfonic acid (PFHxS), Hexadecafluoro-2-
decenoic acid (8:2 FTUCA), and Pentadecafluorodecanoic acid (7:3 FTCA). These eight
chemicals were selected based on interest by stakeholders and aquatic toxicity data availability.

The Clean Water Act (CWA) supports the protection of U.S. waters by helping to ensure
that aquatic species have clean water to live in. States, Tribes, or the EPA may establish
concentration limits of pollutants that will ensure that fish and other aquatic species can live,
grow, and reproduce. CWA Section 304(a) directs the EPA to develop and publish water quality
criteria that reflect the latest scientific knowledge.

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National recommended ambient water quality criteria for the protection of aquatic life
establish chemical concentrations (magnitude) that are averaged over a given time period
(duration) and should not be exceeded more than once during a specified time period
(frequency). The EPA's 304(a)(1) criteria recommendations generally follow the Guidelines for
Deriving Numerical National Water Quality Criteria for the Protection of Aquatic Organisms
and Their Uses (hereafter "Aquatic Life Criteria Guidelines" (U.S. EPA 1985). The Aquatic Life
Criteria Guidelines recommend toxicity data for a minimum of eight families of aquatic animals
be used in developing aquatic life criteria to protect aquatic ecosystems as a whole.

Aquatic life benchmarks, developed under 304(a)(2) of the CWA, are informational
values that the EPA generates when there are limited high quality toxicity data available and data
gaps exist for several families of aquatic organisms. Aquatic life benchmarks provide
information that states and Tribes may consider in their water quality protection programs. In
developing aquatic life benchmarks, data gaps may be filled using new approach methods
(NAMs), such as computer-based toxicity estimation tools (e.g., the EPA's Web-ICE; Version
4.0) or other new approach methods intended to reduce reliance on animal-based testing
(https://www.epa.gov/chemical-research/epa-new-approach-methods-work-plan-reducing-use-
vertebrate-animals-chemical). including the use of read-across estimates based on other
chemicals with similar structures. The EPA's aquatic life benchmark values are not regulatory,
nor do they automatically become part of a state's water quality standards.

Because direct empirical data are more limited for the eight PFAS evaluated within this
document and do not fulfill MDRs as described by the Aquatic Life Criteria Guidelines (U.S.
EPA 1985), the EPA developed acute protective benchmarks for these eight PFAS using
available freshwater species empirical test data in conjunction with the application of the EPA's


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peer-reviewed web-based Interspecies Correlation Estimate (ICE) tool (Web-ICE; Version 4.0;
https://www.epa.gov/webice/) (Raimondo et al. 2010). ICE models support the derivation of
acute values, and do not currently support the derivation of chronic values. This document
accordingly focuses on the derivation of acute aquatic life benchmarks. These acute aquatic life
benchmarks, developed using the direct empirical and ICE (estimated) data for these chemicals,
were calculated by applying methods described in the EPA Aquatic Life Criteria Guidelines
(U.S. EPA 1985).

In addition to addressing data limitations through NAMs, the EPA also developed
application factors to account for species that are known to show markedly greater sensitivity to
PFOA and PFOS. Specifically, Soucek et al. (2023) and Razak et al. (2023) indicated the mayfly,
Neocloeon tricmgulifer, and the cladoceran, Moina micrura, are markedly more sensitive to
PFOA and PFOS relative to other species tested. The EPA accordingly derived the adjustment
factors from the available PFOA and PFOS data and applied them to the eight PFAS benchmarks
to account for greater sensitivity of these species. The inclusion of the application factors protect
the many untested species in the environment, some of which may be similarly sensitive to these
PFAS as the mayfly and cladoceran species noted above. Development of these PFAS
benchmarks reflects goals in the EPA's PFAS Strategic Roadmap and aligns with the Agency's
intention to reduce the use of additional animal testing through application of NAMs. They
provide information to states and Tribes regarding protective values for aquatic life for these
eight data-limited PFAS.

This document describes the process used and results of a systematic review of available
direct empirical toxicity data for aquatic organisms identified via the EPA's literature search for
the eight selected PFAS substances, inclusive of their anionic, acid, and ammonium salt forms.

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The derived acute benchmark values are summarized for each of the eight evaluated PFAS in
Table Ex-1. These concentrations are expected to be protective of 95% of freshwater genera
exposed to the listed PFAS under short term conditions of one-hour of duration, if the one-hour
average magnitude is not exceeded more than once in three years. However, because only limited
toxicity test data were available for these eight chemicals and aquatic toxicity was estimated for
some taxa to develop these benchmarks, the values are somewhat less certain than national
recommended Aquatic Life AWQC developed using more robust empirical data sets (i.e., those
that meet the MDRs according to the EPA's Aquatic Life Criteria Guidelines). Further, while the
eight MDRs were fulfilled for six of the PFAS chemicals when direct empirical and ICE data
were combined, only seven of the eight MDRs were fulfilled for PFBS and only six of the eight
MDRs were fulfilled for PFHxS when data were combined, and there is considered to be greater
uncertainty associated with these benchmark values.

Table Ex-1. Acute Freshwater Aquatic Life Benchmarks for Eight PFAS.

Chemical

PFBA

PFHxA

PFNA

PFDA

PFBS

PFHxS

8:2
FTUCA

7:3
FTC A

Magnitude1

5.3

4.8

0.65

0.50

5.0

0.21

0.037

0.012

Duration

One hour average

Frequency

Not to be exceeded more than once in three years on average

1 Values expressed as mg/L, or ppm.

The EPA additionally explored the application of a data "binning" approach, used by
Giddings et al. (2019) for pyrethroids, as an exploratory approach for deriving protective acute
aquatic life values for grouped perfluoroalkyl carboxylic acids and grouped perfluoroalkyl
sulfonic acids. This approach is based on combining chemicals with similar modes of action to
provide more robust datasets to support value derivation in data-limited situations. The EPA
followed this methodology to explore the calculation of protective benchmark values for six of

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the PFAS, based on the combined perfluoroalkyl carboxylic acid (PFBA, PFHxA, PFNA, PFDA)
and perfluoroalkyl sulfonic acid (PFBS, PFHxS) groupings, that were used to support value
derivation.

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1 Background

This document describes the development of acute water quality benchmarks to protect
freshwater aquatic life from acute toxic effects of eight PFAS substances and their related forms:

•	Perfluorobutanoic acid (PFBA)(CAS# 375224, 45048622, 2218544)

•	Perfluorohexanoic acid (PFHxA)(CAS# 307244, 92612527)

•	Perfluorononanoic acid (PFNA)(CAS# 375951)

•	Perfluorodecanoic acid (PFDA)(CAS# 335762)

•	Perfluorobutanesulfonic acid (PFBS)(CAS# 375735, 108427527, 29420493)

•	Perfluorohexanesulfonic acid (PFHxS)(CAS# 355464, 108427538, 3871996, 82382125)

•	2H-Perfluoro-2-decenoic acid (8:2 FTUCA)(CAS# 70887842)

•	2H,2H,3H,3H-Pefluorodecanoic acid (7:3 FTCA)(CAS# 812704)

These eight chemicals were selected based on interest by stakeholders and aquatic toxicity test
data availability. Notably, the U.S. Department of Defense (DOD) published a set of draft
Ecological Screening Values (Grippo et al. 2021) for six of these substances (PFBA, PFHxA,
PFNA, PFDA, and PFHxS) based on stated concerns about the historic release of these
substances at DOD facilities. The ECOTOXicology Knowledgebase (ECOTOX;
https://cfpub.epa.gov/ecotox/). a curated source of high-quality toxicity data for aquatic life,
terrestrial plants, and wildlife, was also searched to identify PFAS, besides PFOA and PFOS,
that have some of the greatest numbers of empirical studies available to support benchmark
development.

In addition to PFOA and PFOS, for which the EPA has recently developed final Aquatic
Life AWQC, the EPA is aware of thousands of other PFAS chemicals that may be present in the
environment. The above eight chemicals were selected to represent a range of PFAS that are
present in aquatic ecosystems and of concern to stakeholders (e.g., states, Tribes, DOD. Another
important consideration for selection was the availability of both acute direct empirical and ICE
model toxicity data for these chemicals.

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The EPA typically follows the Guidelines for Deriving Numerical National Water
Quality Criteria for the Protection of Aquatic Organisms and Their Uses (U.S. EPA 1985),
which requires toxicity data for a minimum of eight families be available to fulfill taxonomic
MDRs to calculate freshwater Aquatic Life AWQC. The MDR groups represent taxa with
different ecological, trophic, taxonomic and functional characteristics in aquatic ecosystems, and
are intended to collectively represent the diversity and range of sensitivity of species within an
aquatic community. The MDRs, as defined by the Aquatic Life Criteria Guidelines, consist of
the following representative taxa:

a)	Family Salmonidae in the class Osteichthyes

b)	Second family in the class Osteichthyes, preferably a commercially or recreationally
important warmwater species (e.g., bluegill, channel catfish, etc.)

c)	Third family in the phylum Chordata (may be in the class Osteichthyes or may be an
amphibian, etc.)

d)	Planktonic crustacean (e.g., cladoceran, copepod, etc.)

e)	Benthic crustacean (e.g., ostracod, isopod, amphipod, crayfish, etc.)

f)	Insect (e.g., mayfly, dragonfly, damselfly, stonefly, caddisfly, mosquito, midge, etc.)

g)	Family in a phylum other than Arthropoda or Chordata (e.g., Rotifera, Annelida,
Mollusca, etc.)

h)	Family in any order of insect or any phylum not already represented

For some contaminants, particularly those with newer or novel chemistries and emerging
contaminants, these MDRs often cannot be met, and an alternative approach is necessary to
provide information to states and Tribes under CWA Section 304(a). In this case, empirical acute
data for these eight PFAS fulfill some but not all of these MDRs. Accordingly, the EPA applied
the peer reviewed Interspecies Correlation Estimation (ICE) models (Raimondo et al. 2010,
2024) to estimate acute toxicity data to fulfill MDRs for which direct empirical toxicity data are
not available. ICE models support the derivation of acute values, and do not currently support the
derivation of chronic values and/or the values associated with potential bioaccumulative effects.

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This document accordingly focuses on the derivation of acute aquatic life benchmarks. The EPA
has previously used this same approach to derive final acute estuarine/marine benchmarks for
PFOA and PFOS (U.S. EPA 2024b,c).

As noted above, the EPA used the acute empirical test data for these chemicals
supplemented with the ICE values derived for the missing MDRs to derive acute freshwater
benchmark recommendations for aquatic life using calculation procedures provided in the
Aquatic Life Criteria Guidelines (U.S. EPA 1985). The EPA also developed application factors
to account for species that are known to show markedly greater sensitivity to PFOA and PFOS.
Specifically, Soucek et al. (2023) and Razak et al. (2023) indicated the mayfly, Neocloeon
tricmgulifer, and the cladoceran, Moina mi crura, are markedly more sensitive to PFOA and
PFOS relative to other species tested. The EPA accordingly derived the adjustment factors from
the available PFOA and PFOS data and applied them to the eight PFAS benchmarks to account
for greater sensitivity of these species. The inclusion of the application factors protect the many
untested species in the environment, some of which may be similarly sensitive to these PFAS as
the mayfly and cladoceran species noted above. The above approaches are consistent with the
Aquatic Life Criteria Guidelines "good science" clause, the EPA's interest in providing useful
information to states and Tribes regarding protective benchmark values for aquatic life, and also
the EPA's intention to reduce the use of animal testing via application of NAMS in toxicity
assessment (https://www.epa.gov/chemical-research/epa-new-approach-methods-work-plan-
reducing-use-animals-chemical-testing).

The EPA additionally explored the application of a data "binning" approach, used by
Giddings et al. (2019), to derive protective aquatic life values for grouped carboxylic acid PFASs
and grouped sulfonic acid PFAS. This approach, which is detailed in Appendix G, is based on

3


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combining chemicals with similar modes of action to provide more robust datasets to support
value derivation in data-limited situations.

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2 Introduction to ICE

Interspecies Correlation Estimation (ICE) models are log-linear regressions of the acute
toxicity (EC50/LC50) of two species across a range of chemicals that represent the relationship of
inherent sensitivity between those species (Figure 2-1). ICE models are derived from an
extensive, standardized database of acute toxicity values by pairing each species with every other
species for which acceptable toxicity data are available (Raimondo et al. 2010, 2024). ICE
models are used to predict the sensitivity of an untested taxon (predicted taxa are represented by
the y-axis) from the known, measured sensitivity of a surrogate species (represented by the x-
axis).

ICE models have been developed from a broad range of chemicals (e.g., metals and other
inorganics, pesticides, solvents, and reactive chemicals) and across a wide range of toxicity
values and have been validated as accurate predictors of acute toxicity when model criteria
parameters are followed (Raimondo et al. 2024). The Web ICE application (v4.0,
https://www3.epa.gov/webice/) contains approximately 4,800 statistically-significant ICE
models for aquatic animal and plant species.

Validation and uncertainty analyses of ICE models are founded in both leave-one-out
cross validation and direct comparison of measured and predicted acute values (Raimondo et al.
2010, 2024; Willming et al. 2016). Acute toxicity predictions have high accuracy when values
are derived from models with robust parameters (e.g., mean square error and R2 of regression
between the tested and untested species), and variability of prediction accuracy is consistent with
that of empirical test data (Raimondo et al. 2024). Box 1 below outlines the parameter measures
used to define acceptable ICE models for evaluation. Models identified as acceptable based on

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the parameter measures identified in Box 1 were further evaluated for use as discussed in
Sections 4.2 and 5.1.

ICE models have undergone extensive peer review and their use has been supported
through external peer review for development of aquatic life benchmark values for saltwater and
estuarine species exposed to PFOA and PFOS (U.S. EPA 2024b,c). The application of ICE-
predicted values to develop protective aquatic life values by multiple independent, international
groups confirms that values developed from ICE-generated species sensitivity distributions
(SSDs) provide a level of protection that is consistent with using measured laboratory data (Dyer
et al. 2008; Feng et al. 2013; Fojut et al. 2012a,b; Palumbo et al. 2012; Wu et al. 2015, 2016;
Wang et al. 2020; Zhang et al. 2017). A recent review of ICE models additionally supports their
use in regulatory applications based on the reliability of underlying data, model transparency,
statistical robustness, predictive reliability, proof of principle, applicability to probabilistic
approaches, and reproducibility of model accuracy by numerous independent research teams
(Bejarano and Wheeler 2020).

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Figure 2-1. Example of ICE Model for Rainbow Trout (surrogate) and Atlantic Salmon
(predicted).

Each model datapoint is a common chemical that was tested with both species to develop a log-linear
model.

Box I. ICE Model User Parameter Criteria for Robust Predictions
(Willming et al. 2016, Raimondo et al. 2024):

•	Low mean squared error (MSE) (<~ 0.95)

•	High R2 value (>~0.6)

•	High slope (>~0.6; 0.66 - 1.33 when (ig/L input is beyond model domain)

•	Confidence interval range of 2 orders of magnitude between lower and
upper limit

•	For models between vertebrates and invertebrates, using those with lower
MSE or MOA-specific models (not available for PFAS) has been
recommended for listed species predictions

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3 Application of ICE with Selected PFAS

ICE models for freshwater species were used to predict acute toxicity of the eight PFAS
from measured sensitivity of surrogate species listed in Table 3-1. For the application of ICE
models, the type and number of predicted species available to fulfill missing MDRs in the
selected PFAS acute criteria dataset was based on the number of models available within ICE for
the surrogate species (Table 3-1). It is important to note, not all available ICE models met the
prediction robustness criteria outlined in Box 1 for quantitative use. Accordingly, an evaluation
of the predictions was conducted following the acquisition of all available predicted values from
all available surrogate species for each compound.

Table 3-1. Measured Surrogate Species Sensitivity Values and Number of Available ICE
Models for Eight PFAS.

The number of freshwater ICE models for each surrogate excludes models that predict values for species
for which measured acute toxicity data are already available. Available ICE models are presented in

Chemical

Taxa

Surrogate Species

Acute
Toxicity
(mg/L)

Number of
Freshwater ICE
Models

PFBA

Vertebrate

Danio rerio

13,779

18 (5 Juveniles,
13 Embryo)

Invertebrate

Daphnia magna

5,251

69

PFHxA

Invertebrate

Daphnia magna

1,048

69

Vertebrate

Lithobates catesbeianus

1,105

8

PFNA

Invertebrate

Chydorus sphaericus

28

2

Invertebrate

Daphnia magna

85

69

Vertebrate

Xenopus laevis

336

4

PFDA

Invertebrate

Chydorus sphaericus

41

2

Invertebrate

Daphnia magna

120

69

Invertebrate

Daphnia pulicaria

150

2

Vertebrate

Oncorhvnchus mykiss

32

78

Vertebrate

Xenopus laevis

77

4

Invertebrate

Daphnia magna

2,183

69

Vertebrate

Lepomis macrochirus

6,452

76

Vertebrate

Pimephales promelas

1,938

74

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Chemical

Taxa

Surrogate Species

Acute
Toxicity
(mg/L)

Number of
Freshwater ICE
Models

PFHxS

Vertebrate

Danio rerio

23

18 (5 Juveniles,
13 Embryo)

Vertebrate

Lithobates catesbeianus

1,105

8

8:2 FTUCA

Invertebrate

Daphnia magna

3

69

Vertebrate

Oncorhynchus mvkiss

81

78

7:3 FTC A

Invertebrate

Daphnia magna

1

69

Vertebrate

Oncorhynchus mvkiss

32

78

PFAS acute values (typically reported as mg/L) can be beyond the range of values used
to calibrate the ICE model (ICE database toxicity range lxl0"4and lxlO8 (J,g/L) such that the
input PFAS value of the surrogate would be outside the model domain. In situations where the
input PFAS value is beyond the model range, a user can either:

•	Enter the measured toxicity value (LC50/ EC50) into the ICE model as [j,g/L and allow the
regression to extrapolate beyond the model range; or

•	"Scale" the toxicity value by entering the measured LC50 value as mg/L, bringing the
input value within the model range.

Because it is recognized that extrapolating beyond the model range can result in large confidence
intervals around the prediction and can result in the acceptance of fewer ICE models based on
acceptability parameters (see Box 1), a "scaled" approach was considered by Raimondo et al.
(2024) as an alternative approach to predicting species sensitivity to place the input value within
the model domain. The scaled approach underwent detailed evaluation by Raimondo et al.
(2024), who compared the two approaches and reported that ICE models predict toxicity with the
same level of accuracy using scaled and extrapolated input values, with smaller confidence
intervals for the scaled approach (allowing acceptance of more ICE models). The first
("extrapolated") approach was selected as the primary approach for deriving the benchmark

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values. The "scaled" approach was applied as an alternative approach and serves as an additional
analysis to compare SSDs and HC5s with those based on extrapolated values and are
summarized in Section 5.10 and presented in detail in Appendix F.

Figure 3-1 provides an example of the approach taken to calculate a toxicity value when
an LC50 or EC50 value falls beyond the ICE model range, through the use of either an
"extrapolation" or "scaled" approach. The data plots and calculations below provide an
illustrative example using an EC50 value for Daphnia magna for PFBS.

EXTRAPOLATION APPROACH

SCALED APPROACH

Figure 3-1. Example Calculation for Daphnia magna PFBA Toxicity Data Based on
Extrapolation and Scaled Approaches.

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4 Effects Analysis for Aquatic Life

4.1 Summary and Evaluation of Available Empirical Toxicity Test Data

The EPA reviewed and considered all relevant empirical toxicity test data for the eight

PFAS. Information available for all relevant species and genera were reviewed to identify data
from acceptable tests that meet data quality standards. All available empirical studies relating to
the acute toxicological effects of each PFAS on aquatic life were considered in the derivation of
these freshwater acute benchmarks. Empirical data for possible inclusion in the derivation of the
benchmarks were obtained from published literature reporting acute exposures of the selected
PFAS that were associated primarily with mortality/survival. The development of benchmarks
included the review of a range of forms of each PFAS substance for which data were available,
as summarized in Table 4-1. Figure 4-1 presents representative chemical structures for these
PFAS. With the exception of 8:2 FTUCA, all of these PFAS can be analyzed using the EPA
Method 1633 (U.S. EPA 2024a).

Table 4-1. Summary Table of PFAS Forms and Associated CAS Numbers Identified for the
Evaluation of Available Empirical Data.			

PFAS

Form

Chemical Formula

CAS
Number

Perfluorobutanoic acid
(PFBA)

PFBA

C4HF7O2

375224

PFBA Ion

C4F7O2

45048622

PFBA-Na

C4F7Na02

2218544

Perfluorohexanoic Acid
(PFHxA)

PFHxA

c6hf„o2

307244

PFHxA Ion

C6F11O2"

92612527

Perfluorononanoic acid
(PFNA)

PFNA

C9HF17O2

375951

Perfluorodecanoic Acid
(PFDA)

PFDA

C10HF19O2

335762

Perfluorobutanesulfonic acid
(PFBS)

PFBS

C4HF9O3S

375735

T etrabutylammonium
perfluorobutanesulfonate

C20H36F9NO3S

108427527

PFBS.K

C4F9KO3S

29420493

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PFAS

Form

Chemical Formula

CAS
Number

Perfluorohexanesulfonic acid
(PFHxS)

PFHxS

CHF;=()=S

355464

Perfluorohexanesulfonate

C6F1303S

108427538

PFHxS.K

c6f13ko3s

3871996

PFHxS Na

C6Fi3Na03S

82382125

2H-Perfluoro-2-decenoic acid
(8:2 FTUCA)

8:2 FTUCA

C10H2F16O2

70887842

2H,2H,3H,3H-
Pefluorodecanoic acid (7:3
FTCA)

7:3 FTCA

C10H5F15O2

812704

/Ns
HO ^0

Perfluorobutanoic
Acid (PFBA)

o oh

Perfluorohexanoic
Acid (PFHxA)

f f f n

Perfluorononanoic
Acid (PFNA)

Perfluorodecanoic
Acid (PFDA)

O F F F F

OH F F F F



Perfluorobutane
sulfonic Acid
(PFBS)

Perfluorohexane
sulfonic acid
(PFHxS)

2H-Perfluoro-2-
decenoic acid
(8:2 FTUCA)

2H,2H,3H,BH-
Pefluorodecanoic
acid (7:3 FTCA)

Figure 4-1. Chemical Structures of Selected PFAS.

Empirical studies available for the eight PFAS were identified using ECOTOX. Toxicity
studies accessed from the ECOTOX database were further evaluated. Studies were evaluated for

data quality as described by the EPA OW's data quality standard operating procedure (SOP), and

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consistent with OW's data quality review approach (U.S. EPA 1985), and the EPA's Office of
Chemical Safety and Pollution Prevention (OCSPP)'s Ecological Effects Test Guidelines (U.S.
EPA 2016). The EPA completed a Data Evaluation Record (DER) for each species empirical
toxicity test from the studies identified by ECOTOX for the eight PFAS undergoing evaluation.
This in-depth review ensured the studies used for derivation would result in robust, scientifically-
defensible benchmarks. Studies that did not fully meet the data quality objectives outlined in the
EPA SOP were not considered for inclusion in the benchmark derivation, including some studies
with other PFAS exposures. Further, only single chemical toxicity tests (i.e., not tests with
chemical mixtures) were considered for possible inclusion in benchmark derivation. When
available, both controlled laboratory experiments and field observations/studies were evaluated
for inclusion.

Due to the EPA's interest in using all available quality data, particularly for these data-
limited chemicals, toxicity studies were considered for possible inclusion regardless of the test
species residential status in North America. Use of non-North American residential species is
also consistent with other published aquatic life criteria (U.S. EPA 2018). Non-North American
resident species also help to serve as taxonomically-related surrogate organisms for untested
resident species.

Toxicity data from tests reporting either measured or nominal concentrations were
considered for quantitative use in benchmark derivation. Toxicity tests used in many of the
EPA's previous aquatic life criteria documents are typically based only on measured chemical
concentrations. For PFOA and PFOS, the EPA examined the issue of whether nominal
(unmeasured) and measured concentrations are in close agreement with each other (U.S. EPA
2024b,c). For both PFOA and PFOS, pairs of nominal and corresponding measured

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concentrations were compared to one another through: (1) linear correlation analysis and; (2) an
assessment of measured concentrations as a percent of its paired nominal concentration. For
PFOA 83% of measured freshwater concentrations and 82% of the measured concentrations for
PFOS fell within 20% of paired nominal concentrations, which represent the test acceptability
threshold identified by the EPA's OCSPP's Ecological Effects Test Guidelines. Based on the
results for PFOA and PFOS (U.S. EPA 2024b,c), the EPA concluded that nominal test
concentrations adequately represent actual PFAS exposures in standard acute laboratory-based
toxicity tests and PFAS toxicity tests were not excluded from quantitative use in benchmark
derivation on the basis of unmeasured test concentrations alone. Resultingly, renewal, and flow-
through experiments were considered for inclusion for species mean acute values regardless of
whether the PFAS concentrations were measured.

Available empirical acute data meeting the quality objectives and test requirements were
utilized quantitatively in conjunction with ICE values in deriving benchmarks for aquatic life.
The number of acute genus and species level mean values and MDRs fulfilled by the
quantitatively acceptable empirical acute toxicity data available for each of the eight PFAS is
summarized in Table 4-2 and detailed in Appendix A: Quantitatively Acceptable Freshwater
Acute Toxicity Studies. Studies that were determined to be qualitatively acceptable as
supporting information, but not acceptable for quantitative use, are summarized in Appendix C:
Qualitative Freshwater Acute Toxicity Studies. Studies that were not considered acceptable
for quantitative or qualitative use are summarized in Appendix D: Unused Freshwater Acute
Toxicity Studies.

Acute values were presented as reported by the study authors for each individual study.
Author-reported toxicity values were included in each study summary included in corresponding

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appendices, as applicable. The results of all toxicity values (e.g., individual toxicity test
outcomes and species- and genus-mean values) are given to four significant figures to prevent
round-off error in subsequent calculations, not to reflect the precision of the value. The specific
toxicity value utilized in the derivation of the corresponding benchmarks is stated for each study
at the end of the summaries in Appendix A.

Table 4-2. Number of Acute Genus and Species Level Mean Values in the Empirical
Toxicity Datasets and Freshwater Minimum Data Requirements Fulfilled per the Aquatic
Life Criteria Guidelines for the

MDR

PFBA

PFHxA

PPFNA

PFDA

PFBS

PFHxS

8:2
FTUCA

7:3
FTCA

Family Salmonidae in the class
Osteichthyes

-

-

-

1

-

-

1

1

Second family in the class
Osteichthyes, preferably a
commercially or recreationally
important wannwater species

1

-

-

-

3

1

-

-

Third family in the phylum
Chordata (may be in the class
Osteichthyes or may be an
amphibian, etc.)

-

2

1

1

-

2

-

-

Planktonic Crustacean

3

1

2

3

1

-

1

1

Benthic Crustacean

-

-

-

-

-

-

-

-

Insect

-

-

-

-

-

-

-

-

Family in a phylum other than
Arthropoda or Chordata (e.g.,
Rotifera, Annelida, or Mollusca)

-

-

-

-

-

-

-

-

Family in any order of insect or
any phylum not already
represented

1

1

-

-

-

-

-

-

Total number of empirical
studies

5

4

3

5

4

3

2

2

MDRs fulfilled with
empirical data

3

3

2

3

2

2

2

2

4.2 Prediction of Species Sensitivity Using ICE Models

Values for the selected PFAS were first predicted with Web-ICE v4.0 for all possible

species. The resulting ICE models are summarized in Appendix E. These ICE models were then
evaluated using the acceptance parameters (e.g., MSE, R2, slope) summarized in Box 1, provided

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above under Section 2. Only ICE models based on freshwater species were considered for use.
Models that met these acceptance parameters were used to derive species toxicity data, which
were used in conjunction with empirical toxicity data to support the derivation of the benchmark
values. The ICE models selected for use are summarized in each of the chemical-specific
discussions presented in Section 5.

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5 Derivation of Acute Water Quality Benchmarks for Selected
PFAS

5.1 Approach

5.1.1 Use of Empirical and ICE Data

The following section describes the process used to derive the water quality benchmarks

for each of the eight PFAS. The first step encompassed development of the acute toxicity dataset

used to support the derivation of each benchmark, which involved combining available empirical

test data with ICE-derived toxicity data in order to both meet the eight MDRs, as described in the

Aquatic Life Criteria Guidelines (U.S. EPA 1985), and provide as robust of a dataset as possible

for benchmark derivation. The following approach was used to develop the dataset for the

derivation of each acute PFAS benchmark:

•	All acceptable empirical data (evaluated as described in Section 4.1 and summarized in
Appendix A and B) were used and prioritized for benchmark derivation. Accordingly, if
both acceptable empirical and ICE-estimated LCso/ ECso values were available for a
species, then only the empirical data were averaged to derive that SMAV, based on the
greater certainty assumed to be associated with quantitatively-acceptable empirical test
data.

•	ICE-derived toxicity data were derived from acceptable ICE models (as described in
Section 4.2 and summarized in Appendix E), and used to derive toxicity data for each
PFAS, with the objective of fulfilling gaps in the MDRs and creating an overall more
robust acute dataset for benchmark derivation. As noted, ICE-estimated LC50/EC50 values
were used to derive SMAVs, except when empirical data were available. If multiple
acceptable ICE models were available for a given species (based on prediction by

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multiple surrogates) then only the model with the closest taxonomic relationship was
used to support deriving the SMAV.

Following development of the toxicity database, values were then calculated using the
same basic procedure as described in the Aquatic Life Criteria Guidelines (U.S. EPA 1985) for
the derivation of acute criteria. Please refer to the Aquatic Life Criteria Guidelines for a detailed
description of this approach. The primary difference between the derivation of aquatic life
criteria versus the derivation of protective acute benchmarks in this document is in the
composition of the constituent dataset, which consists of both empirical test and ICE-derived
acute toxicity data, since the eight MDRs could not be fulfilled for these compounds using the
available empirical test data.

Consistent with the Aquatic Life Criteria Guidelines (U.S. EPA 1985), acceptable
empirical data were then used in conjunction with ICE estimated LC50 values to derive a
"preliminary" value as follows:

•	Species mean acute values (SMAVs) were derived based on the availability of multiple
acceptable empirical or the selected ICE-estimated values by combining the accepted
data into species-level groupings and calculating the means for each grouping.

•	Genus Mean Acute Values (GMAVs) were then calculated by combining the SMAVs
(comprised of both empirical data and ICE-estimated values) into genus-level groupings
and calculating the means for each grouping.

•	Sensitivity Distributions (SD) were developed from the GMAVs

•	The four GMAVs closest to the 5th percentile of the distribution were used to calculate
the Final Acute Value (FAV).

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•	The FAV is then divided by two, which is intended to estimate a minimal effect level and
provide an acute value that is protective of nearly all individuals in the distribution (U.S.
EPA 1985).

5.1.2 Application Factor to Adjust for Additional Unrepresented Sensitive Species Data

It was noted that no acceptable empirical data and limited ICE-estimated toxicity data

were available for some taxa. This is particularly notable as recent investigations with both

PFOA and PFOS indicate the mayfly, Neocloeon tricmgulifer (Soucek et al. 2023) and the

cladoceran, Moina mi crura (Razak et al. 2023) are among the most acutely sensitive of any

species tested to these substances amongst the acceptable data, as determined by the EPA:

•	For PFOA, the author-reported acute EC so (i.e., 0.4747 mg/L) for M mi crura from a new
study (Razak et al. 2023) was determined to be acceptable for quantitative use and was
averaged as a geometric mean with other quantitatively acceptable acute Moina data,
yielding the most sensitive genus mean acute value (8.885 mg/L). The EPA-calculated
acute LCso (i.e., 13.045 mg/L) for N. triangulifer from Soucek et al. (2023) was
acceptable for quantitative use and was the second most sensitive genus mean acute value
in the final PFOA acute freshwater criterion.

•	For PFOS, the EPA-calculated acute LC50 (i.e., 0.07617 mg/L) for N. triangulifer was
acceptable for quantitative use and was the most sensitive genus mean acute value in the
final PFOS acute freshwater criterion. The new author-reported acute LC50 (i.e., 0.5496
mg/L) forM mi crura (Razak et al. 2023) was acceptable for quantitative use and was
averaged as a geometric mean with other Moina data to yield the second most sensitive
genus mean acute value (3.075 mg/L) in the final acute PFOS criterion.

These data were not available when the draft freshwater PFOA and PFOS acute aquatic life
criteria were released (U.S. EPA 2022a,b), but were incorporated into the final freshwater acute

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criteria released for these compounds (U.S. EPA 2024b,c), significantly lowering the derived
freshwater acute values for these substances:

•	For PFOA, the final acute freshwater criterion is 3.1 mg/L, compared to the draft acute
criterion of 49 mg/L, and the addition of the acute value for N. triangulifer along with the
addition of the sensitive M. micrura data decreased the criterion by a factor of 15.8.

•	For PFOS, the final acute freshwater criterion is 0.071 mg/L, compared to the draft acute
criterion of 3.0 mg/L, and the addition decreased the criterion by a factor of 42.3.

Based on the limited available data and to account for this marked difference in sensitivity, the
EPA applied an application factor to the calculation of each PFAS benchmark value, to account
for the absence of empirical insect, and other sensitive invertebrate, data and the markedly
greater sensitivity of the mayfly N. triangulifer, and M micrura. The following application
factors were used:

•	For PFAS with carboxylate groups (PFBA, PFHxA, PFNA, PFDA, 8:2 FTUCA, and 7:3
FTC A), an application factor of 15.8 was used based on the difference in draft and final
PFOA acute criterion values when the additional sensitive species data were added.

•	For PFAS with sulfonate groups (PFBS and PFHxS), an application factor of 42.3 was
similarly used.

These application factors were used to convert the "preliminary" benchmark value to a final
benchmark value. Table 5-1 provides a summary of the application factors developed to adjust
for insect sensitivity.

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Table 5-1. Application Factors Applied to Adjust for Additional Unrepresented Sensitive
Species Data. 				

Chemical

Draft Acute
Freshwater
Criterion
(mg/L)

Final Acute
Freshwater
Criterion
(mg/L)

Application
Factor

PFAS Applied To

PFOA

49

3.1

15.8

PFBA, PFHxA, PFNA, PFDA,
8:2 FTUCA, 7:3 FTCA

PFOS

3.0

0.071

42.3

PFBS, PFHxS

Final benchmark values were calculated by dividing the value derived from the
sensitivity distribution using procedures presented in the Aquatic Life Criteria Guidelines (U.S.
EPA 1985) by the relevant application factors, to derive benchmark values protective of the
aquatic community, including sensitive aquatic insects. The following sections detail the
derivation of the acute benchmark values for each of the eight PFAS.

5.2 Derivation of Acute Water Benchmark for Perfluorobutanoic Acid
(PFBA)

Quantitatively acceptable empirical data for PFBA were available for five species
comprising four genera and fulfilling three MDR groups (Tables 4-2 and 5-3). Because only
three of the MDRs were met with empirical data, ICE-derived values were used to fulfill the
remaining five MDRs. A total of 22 ICE models were acceptable for use. Of these, three ICE
models were not selected for use because empirical data were available for the three genera (i.e.,
models for Daphniapulex, Chydorus sphaericus and Danio rerio). One other model was not
used because there was an acceptable model for a more closely-related surrogate species (i.e.,
Daphnia magna predicting for Pimephalespromelas). A total of 18 ICE models were selected
for use, resulting in ICE models predicting 18 SMAVs, representing 18 genera and seven MDRs
(Tables 5-2 and 5-3).

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The combined empirical and ICE data resulted in 22 GMAVs that collectively fulfill the
eight MDR groups for deriving a freshwater criterion as defined by the Aquatic Life Criteria
Guidelines (U.S. EPA 1985). The ranked GMAVs for these combined data along with the MDR
met by each GMAV are summarized in Table 5-3. GMAVs for the four most sensitive genera
were within a factor of eight of each other (Table 5-4). The freshwater FAV (the 5th percentile of
the genus sensitivity distribution) for PFBA is 166.1 mg/L (Table 5-4), which is lower than all of
the GMAVs except for one tested species (the rotifer, Brachionus calyciflorus, GMAV=110
mg/L). The FAV was divided by two to obtain a preliminary value of 83.03 mg/L PFBA and
then adjusted by the carboxylic acid application factor (15.8) to obtain the freshwater acute water
column benchmark magnitude of 5.3 mg/L PFBA (rounded to two significant figures). This
value is expected to be protective of 95% of freshwater genera exposed to PFBA under short-
term conditions of one-hour duration, if the one-hour average magnitude is not exceeded more
than once in three years (Figure 5-1).

22


-------
Table 5-2. Acceptable Models for ICE-estimated Species Sensitivity to PFBA.

Includes values that were extrapolated as indicated. Bold predicted ECsos used for SMAV calculations.

Predicted Species

Surrogate

ICE Model
(m

Value Range
B/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Minimum

Maximum

Amphipod (Gammarus fasciatus)

Daplinid (Daplinia magna)

5,251**

0.0002

5,000

878.8

(199.6-3,869.5)

Beaver-tail fairy shrimp (Thamnocephalus
platyurus)

Daplinid (Daplinia magna)

5,251

0.0003

8,694.5

2,248.7

(1,108.2-4,562.9)

Bluegill (Lepomis macrochirus)

Daplinid (Daplinia magna)

5,251

0.0001

46,278.3

959.0

(508.0-1,810.5)

Daplinid (Ceriodaplinia dubia)

Daplinid (Daplinia magna)

5,251

0.0003

46,278.3

4,100.3

(2,125.4-7,909.9)

Daphnid (Daplinia pulex)*

Daplinid (Daplinia magna)

5,251**

0.0002

4,894.7

4,297.8
(2,058.5-8,973.1)

Fathead minnow (Pimephales promelas)

Daplinid (Daplinia magna)

5,251

0.0002

46,500

l,174.4b
(655.0-2,105.8)

Fathead minnow (Pimephales promelas)

Zebrafish-embryo (Danio rerio -
embryo)

13,779

0.023

54,579

11,531.9

(6,189.6-21,485.3)

Fatmucket (Lampsilis siliquoidea)

Daplinid (Daplinia magna)

5,251

0.014

8,694.5

2,013.5

(809.4-5,008.9)

Flagfish (Jordanella floridae)

Zebrafish (Danio rerio)

13,779**

7.797

649.3

85,148.8

(12,990.4-558,127.6)

Goldfish (Carassius auratus)

Zebrafish-embryo (Danio rerio -
embryo)

13,779**

0.145

304.5

19,384.5

(4,544.6-82,682.6)

Midge (Paratanytarsus parthenogeneticus)

Daplinid (Daplinia magna)

5,251

0.37

14,500

12,920.8

(3,394.0-49,189.4)

Oriental river shrimp (Macrobrachium
nipponense)

Daplinid (Daplinia magna)

5,251**

0.011

281.6

7,457.2

(1,357.9-40,952.1)

Paper pondshell (Utterbackia imbecillis)

Daplinid (Daplinia magna)

5,251

0.014

8,694.5

1,748.6

(862.3-3,545.6)

Rainbow trout (Oncorhynchus mykiss)

Daplinid (Daplinia magna)

5,251

0.0001

14,500

677.7

(396.2-1,159.4)

Swamp lymnaea (Lymnaea stagnalis)

Daplinid (Daplinia magna)

5,251

0.014

8,694.5

3,490.7

(670.8-18,164.6)

23


-------
Predicted Species

Surrogate

ICE Model
(m

Value Range

B/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Minimum

Maximum

Tadpole physa (Physella gyrina)

Daphnid (Daphnia magna)

5,251

0.014

8,694.5

3,740.1

(1,061.7-13,176.1)

Threeridge (Amblema plicata)

Daphnid (Daphnia magna)

5,251**

0.014

4,894.7

881.4

(169.6-4,582.0)

Vernal pool fairy shrimp (Branchinecta
lynchi)

Daphnid (Daphnia magna)

5,251

0.014

8,694.5

2,852.6

(949.6-8,569.8)

Washboard (Megalonaias nervosa)

Daphnid (Daphnia magna)

5,251

0.014

8,694.5

1,759.2

(742.4-4,168.8)

Water flea (Chydorus sphaericus)*

Daphnid (Daphnia magna)

5,251**

0.009

977.6

1,522.7
(748.6-3,097.1)

Western pearlshell (Margaritifera falcata)

Daphnid (Daphnia magna)

5,251

0.014

8,694.5

1,528.3

(497.6-4,694.2)

Zebrafish-embryo (Danio rerio-embryo)*

Daphnid (Daphnia magna)

5,251

0.0001

46,500

1,391.4
(413.4-4,683.2)

* Acceptable models that were not used because genus level empirical data were available.

**Measured ECso falls outside range of ICE model and predicted SMAV is based on extrapolation beyond model bounds.
a Saltwater surrogate species were not used.

b Not used because other more closely taxonomically-related models were available.

0 Not used because other more closely taxonomically-related models where the measured surrogate ECso values fell within the ICE model range were available.

24


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Table 5-3. Ranked PFBA Genus Mean Acute Values.

Values in bold and highlighted are derived from empirical PFBA toxicity tests with the species.



MDR





SMAV

GMAV

Percentile

Rank

Group1

Name

Species

(mg/L)

(mg/L)

Rank

1

H

Rotifer

Brachionus calyciflorus

110.0

110.0

0.04

2

A

Rainbow trout

Oncorhvnchus mvkiss

677.7

677.7

0.09

3

E

Amphipod

Gammctrus fascia tus

878.8**

878.8

0.13

4

G

Threeridge

Amblema plicata

881.4**

881.4

0.17

5

B

Bluegill

Lepomis macrochirus

959.0

959.0

0.22

6

G

Western pearlshell

Margaritifera falcata

1,528

1,528

0.26

7

G

Paper pondshell

Utterbackia imbecillis

1,749

1,749

0.30

8

G

Washboard

Megalonaias nervosa

1,759

1,759

0.35

9

G

Fatmucket

Lampsilis siliquoidea

2,013

2,013

0.39

10

E

Beaver-tail fairy
shrimp

Thamnocephalus
platyurus

2,249

2,249

0.43

11

E

Vernal pool fairy
shrimp

Branchinecta lvnchi

2,853

2,853

0.48

12

G

Swamp lymnaea

Lymnaea stagnalis

3,491

3,491

0.52

13

G

Tadpole physa

Physella gyrina

3,740

3,740

0.57

14

D

Daphnid

Ceriodaphnia dubia

4,100

4,100

0.61

15

D

Daphnid

Chydorus sphaericus

>4,281

>4,281

0.65

16

D

Daphnid

Daphnia magna

4,741

4,741

0.70

D

Daphnid

Daphnia pulicaria

>l,006a

17

E

Oriental river shrimp

Macrobrachium
nipponense

7,457**

7,457

0.74

18

B

Fathead minnow

Pimephales promelas

11,532

11,532

0.78

19

F

Midge

Paratanv tarsus
parthenogeneticus

12,921

12,921

0.83

20

B

Zebrafish

Danio rerio

13,779

13,779

0.87

21

B

Goldfish

Carassius auratus

19,385**

19,385

0.91

22

C

Flagfish

Jordanella floridae

85,149**

85,149

0.96

a Not used in GMAV calculation, because values represents a greater than low value

**Measured EC50 falls outside range of ICE model and predicted SMAV is based on extrapolation beyond model

bounds.

1: Freshwater MDR Groups

A)	The family Salmonidae in the class Osteichthyes

B)	A second family in the class Osteichthyes, preferably a commercially or recreationally important

warmwater species (e.g., bluegill, channel catfish, etc.)

C)	A third family in the phylum Chordata (may be in the class Osteichthyes or may be an amphibian, etc.)

D)	A planktonic crustacean (e.g., cladoceran, copepod, etc.)

E)	Abenthic crustacean (e.g., ostracod, isopod, amphipod, crayfish, etc.)

F)	An insect (e.g., mayfly, dragonfly, damselfly, stonefly, caddisfly, mosquito, midge, etc.)

G)	A family in a phylum other than Arthropoda or Chordata (e.g., Rotifera, Annelida, Mollusca, etc.)

H)	A family in any order of insect or any phylum not already represented.

25


-------
Table 5-4. PFBA Protective Aquatic Life Acute Benchmark.

Bold values represent genera for which empirical toxicity data were available.

Calculated Freshwater FAY based on 4 lowest values; N=22 GMAVs total





Benchmark calculated by dividing the FAY by 2 and by the carboxylic acid application factor (15.8)





GMAV









Rank

Genus

(mg/L)

ln(GMAV)

ln(GMAV)2

P=R/(N+1)

sqrt(P)

1

Brachionus

110.0

4.70

22.09

0.043

0.209

2

Oncorhynchns

677.8

6.52

42.49

0.087

0.295

3

Gammarus

878.8

6.78

45.95

0.130

0.361

4

Amblema

881.4

6.78

45.99

0.174

0.417



£ (Sum):

24.78

156.53

0.43

1.28







P = cumulative probability











R = rank













N = number of GMAVs





S2 =

125.08



S = slope







L =

2.612



L = X-axis intercept





A =

5.112



A = InFAV







FAV =

166.1











FAV/2 =

83.03 mg/L (Preliminary Value)







Adjustment =

83.03 / 15.8 = 5.255 (Preliminary Value / Carboxylic Acid Application Factor)

Benchmark =

5.3 mg/L PFBA (rounded to two significant figures)





26


-------
1.00
0.90

g 0.80

CJ

CZ

«

QJ
>

0.70
0.60

x 0.50
a

ft

Pi

£ 0.40 -¦

QJ

5 0.30

Ph

0.20
0.10
0.00

•

Fish (Empirical)

o

Fish (WeblCE)

+

Insect (WeblCE)

~

Invertebrate (Empirical)

A

Invertebrate (WeblCE)

O

Mollusk (WeblCE)



Preliminary Value



- Acute Benchmark

Jordanella O
O Carassius
# Danio
+ Paratanytarsus
O Pimephales
A Macrobrachium
A Daphnia
A Chydorus
A Ceriodaphnia
O Physella
O Lymnaea
A Branchinecta
A Thamnocephalus
O Lampsilis
O Megalonaias
O Utterbackia
O Margaritifera
O Lepomis
O Amblema
A Gammarus
O Oncorhynchus

A Brachionus

10	100	1,000	10,000

Genus Mean Acute Value (mg/L PFBA)

100,000

Figure 5-1. Ranked Acute PFBA GMAVs Used for the Aquatic Life Acute Benchmark
Calculation.

5.3 Derivation of Acute Water Benchmark for Perfluorohexanoic Acid
(PFHxA^

Quantitatively acceptable empirical data for PFHxA were available for four species
comprising three genera and fulfilling three MDR groups (Tables 4-2 and 5-6). Because only
three of the MDRs were met with empirical data, ICE-derived values were used to fulfill the
remaining five MDRs. A total of 28 ICE models were accepted for use. Of these, two ICE
models were not selected for use because empirical data were available for the genus (i.e.,
models for Daphnia magna and Daphniapiilex). Three other ICE models were not used because

27


-------
there was an acceptable model for a more closely-related surrogate species (i.e., Daphnia magna
predicting for Lepomis macrochirus; Daphnia magna predicting for Pimephalespromelas; and
Daphnia magna predicting for Oncorhynchus mykiss). A total of 23 ICE models were selected
for use, resulting in ICE models predicting 23 SMAVs representing, 21 genera, and 6 MDRs
(Tables 5-5 and 5-6).

The combined empirical and ICE data resulted in 24 GMAVs that collectively fulfill the
eight MDR groups for deriving a freshwater criterion as defined by the Aquatic Life Criteria
Guidelines (U.S. EPA 1985). The ranked GMAVs for these combined data along with the MDR
met by each GMAV are summarized in Table 5-6. GMAVs for the four most sensitive genera
were within a factor of 2.4 of each other (Table 5-7). The freshwater FAV (the 5th percentile of
the genus sensitivity distribution) for PFHxA is 150.4 mg/L (Table 5-7), which is lower than all
of the GMAVs except for one tested species, the rotifer, Brachionus calyciflorus, GMAV=140
mg/L). The FAV was divided by two to obtain a preliminary value of 75.21 mg/L PFHxA and
then adjusted by the carboxylic acid application factor (15.8) to obtain the freshwater acute water
column benchmark magnitude of 4.8 mg/L PFHxA (rounded to two significant figures). This
value is expected to be protective of 95% of freshwater genera exposed to PFHxA under short-
term conditions of one-hour duration, if the one-hour average magnitude is not exceeded more
than once in three years (Figure 5-2).

28


-------
Table 5-5. Acceptable Models for ICE-estimated Species Sensitivity of PFHxA.

Includes values that were extrapolated as indicated. Bold predicted EC50S used for SMAY calculations.

Predicted Species

Surrogate

ICE Model
(m

Value Range
2/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Minimum

Maximum

Amphipod (Gammarus fasciatus)

Daplinid (Daplinia magna)

1,048

0.0002

5,000

234.7

(64.5-853.7)

Beaver-tail fairy shrimp
(Thamnocephalus platyurus)

Daplinid (Daplinia magna)

1,048

0.0003

8,694.5

517.3

(280.8-953.1)

Bluegill (Lepomis macrochirus)

Bullfrog (Lithobates
catesbeianus)

1,105**

0.003

233

1,576.9

(395.3-6,289.9)

Bluegill (Lepomis macrochirus)

Daplinid (Daplinia magna)

1,048

0.0001

46,278

303.5b
(176.0-523.2)

Channel catfish (Ictalurus punctatus)

Bullfrog (Lithobates
catesbeianus)

1,105

0.003

20,900

719.8

(362.4-1,429.7)

Daphnid (Ceriodaplinia dubia)

Daplinid (Daplinia magna)

1,048

0.0003

46,278

785.1

(446.2-1,381.6)

Daplinid (Daplinia magna)*

Bullfrog (Lithobates
catesbeianus)

1,105

0.003

20,900

424.1
(68.5-2,627.9)

Daphnid (Daplinia pulex)*

Daplinid (Daplinia magna)

1,048

0.0002

4,894.7

841.4
(444.8-1,591.6)

Fathead minnow (Pimephales
promelas)

Daplinid (Daplinia magna)

1,048

0.0002

46,500

384.7b
(235.8-627.7)

Fathead minnow (Pimephales
promelas)

Bullfrog (Lithobates
catesbeianus)

1,105

0.003

20,900

1,083.6

(468.5-2,506.4)

Fatmucket (Lampsilis siliquoidea)

Daplinid (Daplinia magna)

1,048

0.014

8,694.5

470.8

(220.2-1,006.8)

Goldfish (Carassius auratus)

Bullfrog (Lithobates
catesbeianus)

1,105

0.003

20,900

1,027.7

(405.9-2,602.1)

Midge (Paratanytarsus
parthenogeneticus)

Daplinid (Daplinia magna)

1,048

0.37

14,500

2,676.6

(917.7-7,806.3)

Mosquitofish (Gambusia affinis)

Bullfrog (Lithobates
catesbeianus)

1,105

18.6

20,900

805.1

(394.5-1,643.4)

Neosho mucket (Lampsilis
rafinesqueana)

Daplinid (Daplinia magna)

1,048**

0.042

166.8

404.3

(69.6-2,349.3)

29


-------
Predicted Species

Surrogate

ICE Model
(m

Value Range
2/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Minimum

Maximum

Oriental river shrimp (Macrobrachium
nipponense)

Daphnid (Daplinia magna)

1,048**

0.011

281.6

1,172.2

(290.4-4,732.5)

Paper pondshell (Utterbackia
imbecillis)

Daplinid (Daplinia magna)

1,048

0.014

8,694.5

418.5

(231.7-755.9)

Rainbow trout (Oncorhynchus
mykiss)

Daphnid (Daplinia magna)

1,048

0.0001

14,500

214.2b
(135.8-337.8)

Rainbow trout (Oncorhynchus
mykiss)

Bullfrog (Lithobates
catesbeianus)

1,105

0.003

20,900

713.5

(200.3-2,541.4)

Swamp lymnaea (Lymnaea stagnalis)

Daplinid (Daplinia magna)

1,048

0.014

8,694.5

762.0

(188.8-3,075.8)

Tadpole physa (Physella gyrina)

Daplinid (Daplinia magna)

1,048

0.014

8,694.5

794.5

(277.3-2,276.3)

Threeridge (Amblema plicata)

Daplinid (Daplinia magna)

1,048

0.014

4,894.7

191.3

(48.0-761.8)

Vernal pool fairy shrimp
(Branchinecta lynclii)

Daplinid (Daplinia magna)

1,048

0.014

8,694.5

623.8

(247.0-1,575.4)

Washboard (Megalonaias nervosa)

Daplinid (Daplinia magna)

1,048

0.014

8,694.5

372.3

(181.1-765.2)

Water flea (Chydorus sphaericus)

Daplinid (Daplinia magna)

1,048**

0.009

977.6

329.4

(183.8-590.5)

Wavyrayed lampmussel (Lampsilis
fasciola)

Daplinid (Daplinia magna)

1,048

166.8

4,894.7

686.2

(127.8-3,684.7)

Western pearlshell (Margaritifera
falcata)

Daplinid (Daplinia magna)

1,048

0.014

8,694.5

370.2

(145.8-940.1)

Zebrafish-embryo (Danio rerio-
embryo)

Daplinid (Daplinia magna)

1,048

0.0001

46,500

481.7

(172.4-1,346.1)

*	Acceptable models that were not used because genus level empirical data were available.

*	*Measured ECso falls outside range of ICE model and predicted SMAV is based on extrapolation beyond model bounds.
a Saltwater surrogate species were not used.

b Not used because other more closely taxonomically-related models were available.

0 Not used because other more closely taxonomically-related models where the measured surrogate ECso values fell within the ICE model range were available.

30


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Table 5-6. Ranked PFHxA Genus Mean Acute Values.

Values in bold and highlighted are derived from empirical PFHxA toxicity tests with the species.



MDR





SMAV

GMAV

Percentile

Rank

Group1

Name

Species

(mg/L)

(mg/L)

Rank

1

H

Rotifer

Brachionus calyciflorus

140.0

140.0

0.04

2

G

Threeridge

Amblema plicata

191.3

191.3

0.08

3

E

Amphipod

Gammctrus fascia tus

234.7

234.7

0.12

4

D

Water flea

Chydorus sphaericus

329.4**

329.4

0.16

5

G

Western pearlshell

Margaritifera falcata

370.2

370.2

0.20

6

G

Washboard

Megalonaias nervosa

372.3

372.3

0.24

7

G

Paper pondshell

Utterbackia imbecillis

418.5

418.5

0.28

8

B

Zebrafish

Danio rerio-embrvo

481.7

481.7

0.32



G

Wavyrayed lampmussel

Lampsilis fasciola

686.2





9

G

Neosho mucket

Lampsilis rafinesqueana

404.3**

507.4

0.36



G

Fatmucket

Lampsilis siliquoidea

470.8





10

E

Beaver-tail fairy shrimp

Thamnocephalus
platyurus

517.3

517.3

0.40

11

E

Vernal pool fairy shrimp

Branchinecta lvnchi

623.8

623.8

0.44

12

A

Rainbow trout

Oncorhvnchus mvkiss

713.5

713.5

0.48

13

B

Channel catfish

Ictaliirus piinctatus

719.8

719.8

0.52

14

G

Swamp lymnaea

Lymnaea stagnalis

762.0

762.0

0.56

15

D

Daphnid

Ceriodaphnia dubia

785.1

785.1

0.60

16

G

Tadpole physa

Physella gyrina

794.5

794.5

0.64

17

B

Mosquitofish

Gambusia affinis

805.1

805.1

0.68

18

C

Bullfrog

Lithobates catesbeianus

1,105

915.2

0.72

C

Green frog

Lithobates clamitans

758.0

19

B

Goldfish

Carassins auratus

1,028

1,028

0.76

20

D

Daphnia

Daphnia magna

1,048

1,048

0.80

21

B

Fathead minnow

Pimephales promelas

1,084

1,084

0.84

22

E

Oriental river shrimp

Macrobrachium
nipponense

1,172**

1,172

0.88

23

B

Bluegill

Lepomis macrochirns

1,577**

1,577

0.92

24

F

Midge

Paratanv tarsus
parthenogeneticus

2,677

2,677

0.96

**Measured EC50 falls outside range of ICE model and predicted SMAV is based on extrapolation beyond model
bounds.

1: Freshwater MDR Groups

A)	The family Salmonidae in the class Osteichthyes

B)	A second family in the class Osteichthyes, preferably a commercially or recreationally important

warmwater species (e.g., bluegill, channel catfish, etc.)

C)	A third family in the phylum Chordata (may be in the class Osteichthyes or may be an amphibian, etc.)

D)	A planktonic crustacean (e.g., cladoceran, copepod, etc.)

E)	Abenthic crustacean (e.g., ostracod, isopod, amphipod, crayfish, etc.)

F)	An insect (e.g., mayfly, dragonfly, damselfly, stonefly, caddisfly, mosquito, midge, etc.)

G)	A family in a phylum other than Arthropoda or Chordata (e.g., Rotifera, Annelida, Mollusca, etc.)

H)	A family in any order of insect or any phylum not already represented.

31


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Table 5-7. PFHxA Final Acute Value and Protective Aquatic Acute Benchmark.

Bold values represent genera for which empirical toxicity data were available.	

Calculated Freshwater FAY based on 4 lowest values; N=24 GMAVs total





Benchmark calculated by dividing the FAY by 2 and by the carboxylic acid application factor (15.8)





GMAV









Rank

Genus

(mg/L)

ln(GMAV)

ln(GMAV)2

P=R/(N+1)

sqrt(P)

1

Brachionus

140.0

4.94

24.42

0.040

0.200

2

Amblema

191.3

5.25

27.60

0.080

0.283

3

Gammarus

234.7

5.46

29.79

0.120

0.346

4

Chydorus

329.4

5.80

33.61

0.160

0.400



£ (Sum):

21.45

115.42

0.40

1.23







P = cumulative probability











R = rank













N = number of GMAVs





S2 =

17.41



S = slope







L =

4.080



L = X-axis intercept





A =

5.013



A = InFAV







FAV =

150.4











FAV/2 =

75.21 mg/L (Preliminary Value)







Adjustment =

75.21 / 15.8 = 4.760 (Preliminary Value / Carboxylic Acid Application Factor)

Benchmark =

4.8 mg/L PFHxA (rounded to two significant figures)





32


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1.00
0.90

-a 0.80

o

C5

is

QJ
>

«
3

E

3

u

-£

a
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Pi


-------
not used because there was an acceptable model for a more closely taxonomically-related model
where the measured surrogate EC50 value fell within the ICE model range (i.e., Daphnia magna
predicting for Danio rerio). A total of 24 ICE models were selected for use resulting in ICE
models predicting 24 SMAVs representing 22 genera and 7 MDRs (Tables 5-8 and 5-9).

The combined empirical and ICE data resulted in 25 GMAVs that collectively fulfill the
eight MDR groups for deriving a freshwater criterion as defined by the Aquatic Life Criteria
Guidelines (U.S. EPA 1985). The ranked GMAVs for these combined data along with the MDR
met by each GMAV are summarized in Table 5-9. GMAVs for the four most sensitive genera
were within a factor of 2.0 of each other (Table 5-10). The freshwater FAV (the 5th percentile of
the genus sensitivity distribution) for PFNA is 20.58 mg/L (Table 5-10), which is lower than all
of the GMAVs except for the one ICE-derived species value for the threeridge mussel, Amblema
plicata (GMAV=17.58 mg/L). The FAV was divided by two to obtain a preliminary value of
10.29 mg/L PFNA and then adjusted by the carboxylic acid application factor (15.8) to obtain
the freshwater acute water column benchmark magnitude of 0.65 mg/L PFNA (rounded to two
significant figures). This value is expected to be protective of 95% of freshwater genera exposed
to PFNA under short-term conditions of one-hour duration, if the one-hour average magnitude is
not exceeded more than once in three years (Figure 5-3).

34


-------
Table 5-8. Acceptable Models for ICE-estimated Species Sensitivity to PFNA.

Includes values that were extrapolated as indicated. Bold predicted EC50S used for SMAY calculations.

Predicted Species

Surrogate

ICE Model
(m

Value Range
2/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Minimum

Maximum

Amphipod (Gammarus fasciatus)

Daplinid (Daplinia magna)

84.51

0.0002

5,000

29.8

(10.9-81.6)

Amphipod (Gammarus
pseudolimnaeus)

Daplinid (Daplinia magna)

84.51**

0.0001

68.3

57.2

(12.4-263.1)

Beaver-tail fairy shrimp
(Thamnocephalus platyurus)

Daplinid (Daplinia magna)

84.51

0.0003

8,694.5

52.1

(32.2-84.2)

Bluegill (Lepomis macrochirus)

Daplinid (Daplinia magna)

84.51

0.0001

46,278

50.3

(33.3-75.9)

Daplinid (Ceriodaplinia dubia)

Daplinid (Daplinia magna)

84.51

0.0003

46,278

59.4

(38.2-92.1)

Daplinid (Daphnia magna)*

Water flea (Chydorus
sphaericus)

27.84

0.007

462.0

76.4
(48.0-121.5)

Daplinid (Daplinia pulex)*

Daplinid (Daplinia magna)

84.51

0.0002

4,894.7

65.8
(40.3-107.5)

Daplinid (Daplinia pulicaria)*

Daplinid (Daplinia magna)

84.51

0.014

281.6

84.2
(18.0-392.8)

Daplinid (Simocephalus serrulatus)

Daplinid (Daplinia magna)

84.51**

0.0002

7.2

39.7

(8.4-187.6)

Fathead minnow (Pimephales
promelas)

Daplinid (Daplinia magna)

84.51

0.0002

46,500

67.3

(47.1-96.2)

Fatmucket (Lampsilis siliquoidea)

Daplinid (Daplinia magna)

84.51

0.014

8,694.5

48.6

(27.6-85.6)

Isopod (Asellus aquaticus)

Daplinid (Daplinia magna)

84.51

0.0003

166.8

1,261.8

(217.7-7314.9)

Midge (Paratanytarsus
parthenogeneticus)

Daplinid (Daplinia magna)

84.51

0.37

14,500

228.8

(101.9-513.5)

Neosho mucket (Lampsilis
rafinesqueana)

Daplinid (Daplinia magna)

84.51

0.042

166.8

42.9

(12.1-152.7)

Oligochaete (Limnodrilus
hoffmeisteri)

Daplinid (Daplinia magna)

84.51

0.003

281.6

181.9

(43.1-768.2)

35


-------
Predicted Species

Surrogate

ICE Model
(m

Value Range
2/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Minimum

Maximum

Oriental river shrimp (Macrobrachium
nipponense)

Daphnid (Daplinia magna)

84.51

0.011

281.6

65.1

(24.3-174.3)

Paper pondshell (Utterbackia
imbecillis)

Daphnid (Daplinia magna)

84.51

0.014

8,694.5

44.8

(28.6-70.2)

Pheasantshell (Ortmanniana
pectorosa)

Daplinid (Daplinia magna)

84.51

0.042

545.9

54.4

(13.3-222.4)

Rainbow trout (Oncorhynchus
mykiss)

Daphnid (Daplinia magna)

84.51

0.0001

14,500

35.4

(25.2-49.7)

Swamp lymnaea (Lymnaea stagnalis)

Daplinid (Daplinia magna)

84.51

0.014

8,694.5

70.7

(24.0-207.9)

Tadpole physa (Physella gyrina)

Daplinid (Daplinia magna)

84.51

0.014

8,694.5

70.6

(31.3-159.2)

Threeridge (Amblema plicata)

Daplinid (Daplinia magna)

84.51

0.014

4,894.7

17.6

(6.1-50.8)

Vernal pool fairy shrimp
(Branchinecta lynclii)

Daplinid (Daplinia magna)

84.51

0.014

8,694.5

58.0

(28.6-117.6)

Washboard (Megalonaias nervosa)

Daplinid (Daplinia magna)

84.51

0.014

8,694.5

32.9

(18.5-58.5)

Water flea (Chydorus sphaericus)*

Daplinid (Daplinia magna)

84.51

0.009

977.5

30.1
(19.3-47.1)

Water flea (Moina macrocopa)

Daplinid (Daplinia magna)

84.51

0.009

281.6

44.3

(8.8-223.1)

Western pearlshell (Margaritifera
falcata)

Daplinid (Daplinia magna)

84.51

0.014

8,694.5

40.4

(19.8-82.5)

Zebrafish (Danio rerio)

Daplinid (Daplinia magna)

84.51**

0.0001

50.0

104.2°
(15.4-703.8)

Zebrafish-embryo (Danio rerio -
embryo)

Daplinid (Daplinia magna)

84.51

0.0001

46,500

91.8

(42.1-200.2)

*Acceptable models that were not used because genus level empirical data were available.

* *Measured ECso falls outside range of ICE model and predicted SMAV is based on extrapolation beyond model bounds.
a Saltwater surrogate species were not used.

b Not used because other more closely taxonomically-related models were available.

0 Not used because other more closely taxonomically-related models where the measured surrogate ECso values fell within the ICE model range were available.

36


-------
Table 5-9. Ranked PFNA Genus Mean Acute Values.

Values in bold and highlighted are derived from empirical PFNA toxicity tests with the species.



MDR





SMAV

GMAV

Percentile

Rank

Group1

Name

Species

(mg/L)

(mg/L)

Rank

1

G

Threeridge

Amblema plicata

17.58

17.58

0.04

2

D

Daphnid

Chydorus sphaericus

27.84

27.84

0.08

3

G

Washboard

Megalonaias nervosa

32.90

32.90

0.12

4

A

Rainbow trout

Oncorhvnchus mvkiss

35.40

35.40

0.15

5

D

Daphnid

Simocephalus serrulatus

39.74**

39.74

0.19

6

G

Western pearlshell

Margaritifera falcata

40.41

40.41

0.23



E

Amphipod

Gammctrus fascia tus

29.82





7

E

Amphipod

Gammarus
pseudolimnaeus

57.16**

41.28

0.27

8

D

Water flea

Moina macrocopa

44.31

44.31

0.31

9

G

Paper pondshell

Utterbackia imbecillis

44.82

44.82

0.35

10

G

Neosho mucket

Lampsilis rafinesqueana

42.93

45.69

0.38

G

Fatmucket

Lampsilis siliquoidea

48.63

11

B

Bluegill

Lepomis macrochirns

50.29

50.29

0.42

12

E

Beaver-tail fairy
shrimp

Thamnocephalus
platyurus

52.08

52.08

0.46

13

G

Pheasantshell

Ortmanniana pectorosa

54.41

54.41

0.50

14

E

Vernal pool fairy
shrimp

Branchinecta lvnchi

58.02

58.02

0.54

15

D

Daphnid

Ceriodaphnia dubia

59.35

59.35

0.58

16

E

Oriental river shrimp

Macrobrachium
nipponense

65.10

65.10

0.62

17

B

Fathead minnow

Pimephales promelas

67.29

67.29

0.65

18

G

Tadpole physa

Physella gyrina

70.62

70.62

0.69

19

G

Swamp lymnaea

Lymnaea stagnalis

70.68

70.68

0.73

20

D

Daphnid

Daphnia magna

84.51

84.51

0.77

21

B

Zebrafish

Danio rerio

91.85

91.85

0.81

22

H

Oligochaete

Limnodrilus hoffmeisteri

181.9

181.9

0.85

23

F

Midge

Paratanv tarsus
parthenogeneticus

228.8

228.8

0.88

24

C

Clawed frog

Xenopus sp.

335.8

335.8

0.92

25

E

Isopod

Asellus aquaticus

1,262

1,262

0.96

**Measured EC50 falls outside range of ICE model and predicted SMAV is based on extrapolation beyond model

bounds.

1: Freshwater MDR Groups

A)	The family Salmonidae in the class Osteichthyes

B)	A second family in the class Osteichthyes, preferably a commercially or recreationally important

warmwater species (e.g., bluegill, channel catfish, etc.)

C)	A third family in the phylum Chordata (may be in the class Osteichthyes or may be an amphibian, etc.)

D)	A planktonic crustacean (e.g., cladoceran, copepod, etc.)

E)	Abenthic crustacean (e.g., ostracod, isopod, amphipod, crayfish, etc.)

F)	An insect (e.g., mayfly, dragonfly, damselfly, stonefly, caddisfly, mosquito, midge, etc.)

37


-------
G)	A family in a phylum other than Arthropoda or Chordata (e.g., Rotifera, Annelida, Mollusca, etc.)

H)	A family in any order of insect or any phylum not already represented.

Table 5-10. PFNA Final Acute Value and Protective Aquatic Acute Benchmark.

Bold values represent genera for which empirical toxicity data were available.	

Calculated Freshwater FAY based on 4 lowest values; N=25 GMAVs total





Benchmark calculated by dividing the FAY by 2 and by the carboxylic acid application factor (15.8)





GMAV









Rank

Genus

(mg/L)

ln(GMAV)

ln(GMAV)2

P=R/(N+1)

sqrt(P)

1

Amblema

17.58

2.87

8.22

0.038

0.196

2

Chydorus

27.84

3.33

11.07

0.077

0.277

3

Megalonaias

32.90

3.49

12.20

0.115

0.340

4

Oncorhynchns

35.40

3.57

12.72

0.154

0.392



£ (Sum):

13.25

44.21

0.38

1.21







P = cumulative probability











R = rank













N = number of GMAVs





S2 =

13.85



S = slope







L =

2.192



L = X-axis intercept





A =

3.024



A = InFAV







FAV =

20.58











FAV/2 =

10.29 mg/L (Preliminary Value)







Adjustment =

10.29 / 15.8 = 0.6511 (Preliminary Value / Carboxylic Acid Application Factor)

Benchmark =

0.65 mg/L PFNA (rounded to two significant figures)





38


-------
JX
a
ss
C4


-------
there was an acceptable model for a more closely-related surrogate species (i.e., Daphnia magna
predicting for Lepomis macrochirus\ Daphnia magna predicting for Pimephalespromelas; and
Daphnia magna predicting for Danio rerio). A total of 46 ICE models were selected for use
resulting in ICE models predicting 46 SMAVs representing 38 genera and all eight MDRs
(Tables 5-11 and 5-12)

The combined empirical and ICE data resulted in 42 GMAVs that collectively fulfill the
eight MDR groups for deriving a freshwater criterion as defined by the Aquatic Life Criteria
Guidelines (U.S. EPA 1985). The ranked GMAVs for these combined data along with the MDR
met by each GMAV are summarized in Table 5-12. GMAVs for the four most sensitive genera
were within a factor of 2.9 of each other (Table 5-13). The freshwater FAV (the 5th percentile of
the genus sensitivity distribution) for PFDA is 15.71 mg/L (Table 5-13), which is lower than all
of the GMAVs except for the ICE-derived species values for the isopod, Caecidotea brevicauda,
GMAV= 8.17 mg/L. The FAV was divided by two to obtain a benchmark value of 7.856 mg/L
PFDA and then adjusted by the carboxylic acid application factor (15.8) to obtain the freshwater
acute water column benchmark magnitude of 0.50 mg/L PFDA (rounded to two significant
figures). This value is expected to be protective of 95% of freshwater genera exposed to PFDA
under short-term conditions of one-hour duration, if the one-hour average magnitude is not
exceeded more than once in three years (Figure 5-4).

40


-------
Table 5-11. Acceptable Models for ICE-estimated Species Sensitivity to PFDA.

Includes values that were extrapolated as indicated. Bold predicted EC50S used for SMAY calculations.

Predicted Species

Surrogate

ICE Model
(m

Value Range
2/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Minimum

Maximum

Amphipod (Gammarus fasciatus)

Daplinid (Daplinia magna)

119.7

0.0002

5,000

39.7

(14.0-112.7)

Amphipod (Gammarus
pseudolimnaeus)

Daplinid (Daplinia magna)

119.7**

0.0001

68.3

78.6

(16.1-384.0)

Apache trout (Oncorhynchus gilae)*

Rainbow trout (Oncorhynchus
mykiss)

32**

0.0041

1.625

39.4
(9.0-173.4)

Atlantic salmon (Salmo salar)

Rainbow trout (Oncorhynchus
mykiss)

32

0.0002

95.86

31.0

(15.6-61.3)

Beaver-tail fairy shrimp
(Thamnocephalus platyurus)

Daplinid (Daplinia magna)

119.7

0.0003

8,694.5

71.5

(43.6-117.5)

Bluegill (Lepomis macrocliirus)

Rainbow trout (Oncorhynchus
mykiss)

32

0.0001

8,341.5

33.1

(27.6-39.6)

Bluegill (Lepomis macrocliirus)

Daplinid (Daplinia magna)

119.7

0.0001

46,278

64.5b
(42.0-99.0)

Bonytail (Gila elegans)

Rainbow trout (Oncorhynchus
mykiss)

32

0.003

43.1

72.7

(12.6-420.2)

Brook trout (Salvelinus fontinalis)

Rainbow trout (Oncorhynchus
mykiss)

32

0.0006

96.7

31.4

(18.8-52.5)

Brown trout (Salmo trutta)

Rainbow trout (Oncorhynchus
mykiss)

32

0.0002

95.9

33.1

(19.8-55.3)

Bullfrog (Lithobates catesbeianus)

Rainbow trout (Oncorhynchus
mykiss)

32

0.0006

13,400

58.8

(23.7-145.4)

Cape Fear shiner (Notropis
mekistocholas)

Rainbow trout (Oncorhynchus
mykiss)

32**

0.004

1.625

114.0

(33.0-394.0)

Channel catfish (Ictalurus punctatus)

Rainbow trout (Oncorhynchus
mykiss)

32

<0.0001

13,400

31.6

(19.9-50.1)

Chinook salmon (Oncorhynchus
tshawytscha)*

Rainbow trout (Oncorhynchus
mykiss)

32

0.003

724.4

40.0
(24.0-66.9)

Coho salmon (Oncorhynchus
kisutch)*

Rainbow trout (Oncorhynchus
mykiss)

32

<0.0001

43.1

49.4
(38.4-63.6)

41


-------
Predicted Species

Surrogate

ICE Model
(m

Value Range
2/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Minimum

Maximum

Common carp (Cyprinus carpio)

Rainbow trout (Oncorhynchus
mykiss)

32

0.0002

198.4

35.6

(16.3-77.7)

Cutthroat trout (Oncorhynchus
clarkii)*

Rainbow trout (Oncorhynchus
mykiss)

32

<0.0001

198.4

27.1
(18.7-39.3)

Cuvier's foam froglet (Physalaemus
cuvieri)

Rainbow trout (Oncorhynchus
mykiss)

32

0.71

155.5

51.8

(30.8-87.1)

Daplinid (Ceriodaplinia dubia)

Daplinid (Daplinia magna)

119.7

0.0003

46,278

84.8

(53.8-133.7)

Daplinid (Daphnia magna)*

Water flea (Chydorus
sphaericus)

41.13

0.007

462.0

114.5
(70.9-184.8)

Daplinid (Daplinia magna)*

Daplinid (Daplinia pulicaria)

149.6

0.009

237.4

122.1
(27.4-544.5)

Daplinid (Daplinia pulex)*

Daplinid (Daplinia magna)

119.7

0.0002

4,894.7

93.7
(56.2-156.0)

Daplinid (Daplinia pulicaria)*

Daplinid (Daplinia magna)

119.7

0.014

281.6

122.0
(24.2-614.6)

Daplinid (Simocephalus serrulatus)

Daplinid (Daplinia magna)

119.7**

0.0002

7.2

55.1

(11.0-275.1)

Fathead minnow (Pimephales
promelas)

Rainbow trout (Oncorhynchus
mykiss)

32

0.0002

13,400

46.7

(36.0-60.7)

Fathead minnow (Pimephales
promelas)

Daplinid (Daplinia magna)

119.7

0.0002

46,500

85.6b
(58.9-124.4)

Fatmucket (Lampsilis siliquoidea)

Daplinid (Daplinia magna)

119.7

0.014

8,694.5

66.6

(37.0-119.8)

Goldfish (Carassius auratus)

Rainbow trout (Oncorhynchus
mykiss)

32

<0.0001

13,400

62.2

(31.6-122.5)

Green sunfish (Lepomis cyanellus)

Rainbow trout (Oncorhynchus
mykiss)

32**

<0.0001

19.9

45.8

(19.9-105.3)

Guppy (Poecilia reticulata)

Rainbow trout (Oncorhynchus
mykiss)

32

0.0006

198.4

28.3

(7.6-105.3)

Isopod (Asellus aquaticus)

Daplinid (Daplinia magna)

119.7

0.0003

166.8

1,699.9

(273.7-10,558.5)

Isopod (Caecidotea brevicauda)

Rainbow trout (Oncorhynchus
mykiss)

32

0.0006

82.0

8.2

(1.9-35.7)

42


-------
Predicted Species

Surrogate

ICE Model
(m

Value Range
2/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Minimum

Maximum

Lake trout (Salvelinus namaycush)

Rainbow trout (Oncorhynchus
mykiss)

32

0.0002

198.4

18.0

(11.4-28.3)

Largemouth bass (Micropteras
salmoides)

Rainbow trout (Oncorhynchus
mykiss)

32

<0.0001

95.9

19.5

(9.5-40.3)

Medaka (Oryzias latipes)

Rainbow trout (Oncorhynchus
mykiss)

32

0.007

54.4

52.1

(10.9-248.7)

Midge (Paratanytarsus dissimilis)

Rainbow trout (Oncorhynchus
mykiss)

32

0.0006

1,330

105.8

(35.6-314.4)

Midge (Paratanytarsus
parthenogeneticus)

Daphnid (Daphnia magna)

119.7

0.37

14,500

321.4

(140.5-735.3)

Neosho mucket (Lampsilis
rafinesqueana)

Daphnid (Daphnia magna)

119.7

0.042

166.8

58.5

(15.4-222.1)

Northern leopard frog (Lithobates
pipiens)

Rainbow trout (Oncorhynchus
mykiss)

32

0.002

198.4

64.7

(33.7-124.2)

Oligochaete (Limnodrilus
hoffmeisteri)

Daphnid (Daphnia magna)

119.7

0.003

281.6

232.8

(51.5-1,052.2)

Oriental river slirimp (Macrobrachium
nipponense)

Daphnid (Daphnia magna)

119.7

0.011

281.6

97.1

(34.6-272.8)

Paper pondshell (Utterbackia
imbecillis)

Daphnid (Daphnia magna)

119.7

0.014

8,694.5

61.0

(38.3-97.2)

Pheasantshell (Ortmanniana
pectorosa)

Daphnid (Daphnia magna)

119.7

0.042

545.9

76.0

(17.4-332.2)

Rainbow trout (Oncorhynchus
mykiss)*

Daphnid (Daphnia magna)

119.7

0.0001

14,500

45.4
(31.9-64.7)

Razorback sucker (Xyrauchen
texanus)

Rainbow trout (Oncorhynchus
mykiss)

32

0.003

43.1

26.6

(5.3-134.0)

Shortnose sturgeon (Acipenser
brevirostruin)

Rainbow trout (Oncorhynchus
mykiss)

32

0.03

95.9

45.7

(11.2-185.6)

Silver perch (Bidyanus bidyanus)

Rainbow trout (Oncorhynchus
mykiss)

32**

0.0014

7.075

73.0

(29.7-179.7)

Sockeye salmon (Oncorhynchus
nerka)*

Rainbow trout (Oncorhynchus
mykiss)

32**

0.03

7.5

25.7
(4.0-164.6)

Southern leopard frog (Lithobates
sphenocephaly)

Rainbow trout (Oncorhynchus
mykiss)

32**

0.03

9.7

312.8

(88.4-1,106.8)

43


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Predicted Species

Surrogate

ICE Model
(m

Value Range
2/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Minimum

Maximum

Swamp lymnaea (Lymnaea stagnalis)

Daplinid (Daphnia magna)

119.7

0.014

8,694.5

98.2

(32.2-299.3)

Tadpole physa (Physella gyrina)

Daplinid (Daplinia magna)

119.7

0.014

8,694.5

98.7

(42.7-227.9)

Threeridge (Amblema plicata)

Daplinid (Daplinia magna)

119.7

0.014

4,894.7

24.5

(8.2-73.1)

Vernal pool fairy shrimp
(Branchinecta lynchi)

Daplinid (Daplinia magna)

119.7

0.014

8,694.5

80.6

(38.8-167.4)

Washboard (Megalonaias nervosa)

Daplinid (Daplinia magna)

119.7

0.014

8,694.5

46.0

(25.6-82.8)

Water flea (Chydorus sphaericus)*

Daplinid (Daplinia magna)

119.7

0.009

977.6

41.9
(26.5-66.4)

Water flea (Moina macrocopa)

Daplinid (Daplinia magna)

119.7

0.009

281.6

56.9

(10.6-305.9)

Western pearlshell (Margaritifera
falcata)

Daplinid (Daplinia magna)

119.7

0.014

8,694.5

54.9

(26.3-114.5)

Yellow perch (Perca flavescens)

Rainbow trout (Oncorhynchus
mykiss)

32**

<0.0001

16.2

22.2

(5.1-95.7)

Zebrafish (Danio rerio)

Rainbow trout (Oncorhynchus
mykiss)

32

0.0002

229.1

17.6

(6.4-48.5)

Zebrafish-embryo (Danio rerio -
embryo)

Rainbow trout (Oncorhynchus
mykiss)

32

0.0002

8,341.5

34.4

(16.8-70.4)

Zebrafish-embryo (Danio rerio-
embryo)

Daplinid (Daplinia magna)

119.7

0.0001

46,500

115.5b
(51.4-259.4)

*Acceptable models that were not used because genus level empirical data were available.

**Measured ECso falls outside range of ICE model and predicted SMAV is based on extrapolation beyond model bounds.
a Saltwater surrogate species were not used.

b Not used because other more closely taxonomically-related models were available.

0 Not used because other more closely taxonomically-related models where the measured surrogate ECso values fell within the ICE model range were available.

44


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Table 5-12. Ranked PFDA Genus Mean Acute Values.

Values in bold and highlighted are derived from empirical PFDA toxicity tests with the species.

Rank

MDR
Group1

Name

Species

SMAV
(mg/L)

GMAV
(mg/L)

Percentile
Rank

1

E

Isopod

Caecidotect brevicauda

8.173

8.173

0.02

2

B

Largemouth bass

Micropterns scdmoides

19.52

19.52

0.05

3

B

Yellow perch

Percct flavescens

22.20**

22.20

0.07

4

A

Brook trout

Salve linns fontinalis

31.44

23.80

0.09

A

Lake trout

Salvelinus namaycush

18.01

5

G

Threeridge

Amblema plicata

24.46

24.46

0.12

6

B

Zebrafish

Danio rerio

24.56

24.56

0.14

7

B

Razorback sucker

Xvrctuchen texamis

26.57

26.57

0.16

8

B

Guppy

Poecilia reticulata

28.31

28.31

0.19

9

B

Channel catfish

Ictalurus punctatiis

31.55

31.55

0.21

10

A

Rainbow trout

Oncorhynchus mykiss

32.00

32.00

0.23

11

A

Atlantic salmon

Salmo salar

30.96

32.02

0.26

A

Brown trout

Salmo trutta

33.12

12

B

Common carp

Cyprinus carpio

35.58

35.58

0.28

13

B

Green sunfish

Lepomis cyanellus

45.79**

38.92

0.30

B

Bluegill

Lepomis macrochirns

33.08

14

D

Daphnid

Chydorus sphaericus

41.13

41.13

0.33

15

B

Shortnose sturgeon

Acipenser brevirostrum

45.67

45.67

0.35

16

G

Washboard

Megalonaias nervosa

46.01

46.01

0.37

17

B

Fathead minnow

Pimephales promelas

46.72

46.72

0.40

18

C

Cuvier's foam froglet

Phvsalaemus cuvieri

51.77

51.77

0.42

19

B

Medaka

Oryzias latipes

52.12

52.12

0.44

20

G

Western pearlshell

Margaritifera falcata

54.89

54.89

0.47

21

D

Daphnid

Simocephalus serndatus

55.07**

55.07

0.49

22

E

Amphipod

Gammarus fasciatas

39.66

55.84

0.51

E

Amphipod

Gammarus
pseudolimnaeus

78.62**

23

D

Water flea

Moina macrocopa

56.93

56.93

0.53

24

G

Paper pondshell

Utterbackia imbecillis

61.04

61.04

0.56

25

B

Goldfish

Carassins auratus

62.18

62.18

0.58

26

G

Neosho mucket

Lampsilis raflnesqueana

58.54

62.42

0.60

G

Fatmucket

Lampsilis siliquoidea

66.56

27

E

Beaver-tail fairy
shrimp

Thamnocephalus
platyurus

71.54

71.54

0.63

28

B

Bonytail

Gila elegans

72.68

72.68

0.65

29

B

Silver perch

Bidyanns bidyanns

73.03**

73.03

0.67

30

G

Pheasantshell

Ortmanniana pectorosa

76.02

76.02

0.70

31

C

Clawed frog

Xenopus sp.

76.50

76.50

0.72

45


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MDR





SMAV

GMAV

Percentile

Rank

Group1

Name

Species

(mg/L)

(mg/L)

Rank

32

E

Vernal pool fairy
shrimp

Brcmchinecta lvnchi

80.58

80.58

0.74

33

D

Daphnid

Ceriodaphnia dabia

84.82

84.82

0.77

34

E

Oriental river shrimp

Macrobrachium
nipponense

97.09

97.09

0.79

35

G

Swamp lymnaea

Lymnaea stagnalis

98.19

98.19

0.81

36

G

Tadpole physa

Physella gyrina

98.69

98.69

0.84



C

Bullfrog

Lithobates catesbeianus

58.75





37

C

Northern leopard frog

Lithobates pipiens

64.72

106.0

0.86

C

Southern leopard frog

Lithobates
sphenocephalus

312.8**

38

B

Cape Fear shiner

Notropis mekistocholas

114.0**

114.0

0.88

39

D

Daphnid

Daphnia magna

145.5

147.5

0.91

D

Daphnid

Daphnia pulicaria

149.6

40

F

Midge

Paratanv tarsus
dissimilis

105.8

184.4

0.93

F

Midge

P aratanv tarsus
parthenogeneticus

321.4

41

H

Oligochaete

Limnodrilus hoffmeisteri

232.8

232.8

0.95

42

E

Isopod

Asellus aquaticus

1,700

1,700

0.98

**Measured EC50 falls outside range of ICE model and predicted SMAV is based on extrapolation beyond model
bounds.

1: Freshwater MDR Groups

A)	The family Salmonidae in the class Osteichthyes

B)	A second family in the class Osteichthyes, preferably a commercially or recreationally important

warmwater species (e.g., bluegill, channel catfish, etc.)

C)	A third family in the phylum Chordata (may be in the class Osteichthyes or may be an amphibian, etc.)

D)	A planktonic crustacean (e.g., cladoceran, copepod, etc.)

E)	Abenthic crustacean (e.g., ostracod, isopod, amphipod, crayfish, etc.)

F)	An insect (e.g., mayfly, dragonfly, damselfly, stonefly, caddisfly, mosquito, midge, etc.)

G)	A family in a phylum other than Arthropoda or Chordata (e.g., Rotifera, Annelida, Mollusca, etc.)

H)	A family in any order of insect or any phylum not already represented.

46


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Table 5-13. PFDA Final Acute Value and Protective Aquatic Acute Benchmark.

Calculated Freshwater FAY based on 4 lowest values; N=42 GMAVs total





Benchmark calculated by dividing the FAY by 2 and by the carboxylic acid application factor (15.8)





GMAV









Rank

Genus

(mg/L)

ln(GMAV)

ln(GMAV)2

P=R/(N+1)

sqrt(P)

1

Caecidotea

8.17

2.10

4.41

0.023

0.152

2

Micropterus

19.52

2.97

8.83

0.047

0.216

3

Perca

22.20

3.10

9.61

0.070

0.264

4

Salve limis

23.80

3.17

10.05

0.093

0.305



£ (Sum):

11.34

32.90

0.23

0.94







P = cumulative probability











R = rank













N = number of GMAVs





S2 =

57.22



S = slope







L =

1.063



L = X-axis intercept





A =

2.754



A = InFAV







FAV =

15.71











FAV/2 =

7.856 mg/L (Preliminary Value)







Adjustment =

7.856 / 15.8 = 0.4972 (Preliminary Value / Carboxylic Acid Application Factor)

Benchmark =

0.50 mg/L PFDA (rounded to two significant figures)





47


-------
1.00 -T
0.90

g 0.80 4





s

3

u

0.70
0.60
0.50

u
a

rt

Pi

jj 0.40
a

| 0.30 4

Oh

0.20
0.10
0.00

0.1

¦

Amphibian (Empirical)

~

Amphibian (WeblCE)

•

Fish (Empirical)

o

Fish (WeblCE)

+

Insect (WeblCE)

~

Invertebrate (Empirical)

A

Invertebrate (WeblCE)

O

Mollusk (WeblCE)



Preliminary Value



-Acute Benchmark

A Limnodrilus
+ Paratanytarsus
~ Daphnia
O Notropis
~ Lithobates
O Physella
O Lvmnaea
A Macrobrachium
A Ceriodaphnia
A Branchinecta
¦ Xenopus
O Ortmanniana
O Bidvanus
O Gila

A Thamnocephalus
O Lampsilis
O Carassius
O Utterbackia
A Moina
A Gammarus
A Simocephalus
O Margaritifera
O Orvzias
~ Physalaemus
O Pimephales
O Megalonaias
O Acipenser

A Asellus

A Chvdorus
O Lepomis
O Cyprinus
O Salmo
# Oncorhvnchus
O Ictalurus
O Poecilia
O Xyrauchen
O Danio
O Amblema
O Sah'elinus
O Perca
O Micropterus
Caecidotea

	i	i	i	¦ ¦ ¦ ¦ ¦ ¦	

10

100

1000

10000

Genus Mean Acute Value (mg/L PFDA)

Figure 5-4. Ranked Acute PFDA GMAVs Used for the Aquatic Life Acute Benchmark
Calculation.

5.6 Derivation of Acute Water Benchmark for Perfluorobutanesulfonie
Acid (PFBS)

Quantitatively acceptable empirical data for PFBS were available for four species
comprising four genera and fulfilling two MDR groups (Tables 4-2 and 5-15). Because only two
of the MDRs were met with empirical data, ICE-derived values were used to fulfill the remaining
six MDRs. A total of 55 ICE models were acceptable for use. Of these, thirteen ICE models were
not selected for use because empirical data were available for the genera (e.g., models for
Lepomis macrochirus and Daphnia magna). Three ICE models were not selected for use because

48


-------
they were derived based on saltwater species surrogates (i.e., model for an amphipod, Hyalella
azteca; daphnid, Ceriodaphnia dubia; and rainbow trout, Oncorhynchus mykiss). Four other
models were not used because there were acceptable models for more closely-related surrogate
species (i.e., Lepomis macrochirus predicting for Cyprinus carpio; Lepomis macrochirus
predicting for Carassius auratus; Daphnia magna predicting for Oncorhynchus mykiss; and
Pimephales promelas predicting for Amble ma plicata). Two additional models (models for the
bullfrog, Lithobates catesbeianus and channel catfish, Ictaluruspunctatus) were not used
because the measured EC50 values fell outside of the ICE model range and other acceptable
models for taxonomically-related species with measured EC50 values within the ICE model range
were used. A total of 33 ICE models were selected for use, resulting in ICE models predicting 30
SMAVs representing 26 genera and seven MDRs (Tables 5-14 and 5-15). An ICE model was
not available to fulfill the MDR for a family in any order of insect or any phylum not already
represented (Tables 5-14 and 5-15).

The combined empirical and ICE data resulted in 30 GMAVs that collectively fulfill
seven MDR groups for deriving a freshwater criterion as defined by the Aquatic Life Criteria
Guidelines (U.S. EPA 1985). The ranked GMAVs for these combined data along with the MDR
met by each GMAV are summarized in Table 5-15. Surrogate species EC50 values fell outside
the ICE model range for 14 of the predicted SMAVs and the resultant GMAVs are less certain
than GMAVs based on ICE models where surrogate EC50 values were in the range of the model.
GMAVs for the four most sensitive genera were within a factor of 1.7 of each other (Table 5-
16). The freshwater FAV (the 5th percentile of the genus sensitivity distribution) for PFBS is
422.1 mg/L (Table 5-16), which is lower than all of the GMAVs except for the ICE-derived
species value for the threeridge mussel, Amblemaplicata (GMAV=383.6 mg/L). The FAV was

49


-------
divided by two to obtain a preliminary value of 211.0 mg/L PFBS and then adjusted by the
sulfonic acid application factor (42.3) to obtain the freshwater acute water column benchmark
magnitude of 5.0 mg/L PFBS (rounded to two significant figures). This value is expected to be
protective of 95% of freshwater genera exposed to PFBS under short-term conditions of one-
hour duration, if the one-hour average magnitude is not exceeded more than once in three years
(Figure 5-5).

50


-------
Table 5-14. Acceptable Models for ICE-estimated Species Sensitivity to PFBS.

Includes values that were extrapolated as indicated. Bold predicted EC50S used for SMAY calculations.

Predicted Species

Surrogate

ICE Model
(m

Value Range
B/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Minimum

Maximum

Amphipod (Gammarus fasciatus)

Daplinid (Daplinia magna)

2,183

0.0002

5,000

428.1

(107.9-1,697.9)

Amphipod (Hyalella azteca)

Mysid (Americamysis baliia)

372**

<0.0001

193.4

4494a

(93.4-2,162.8)

Atlantic salmon (Salmo salar)

Bluegill (Lepomis
macrocliirus)

6,452**

0.004

123.3

14,523.6

(2,825.4-74,655)

Beaver-tail fairy shrimp
(Thamnocephalus platyurus)

Daplinid (Daplinia magna)

2,183

0.0003

8,694.5

1,010

(525.1-1,942.9)

Bluegill (Lepomis macrocliirus)*

Daplinid (Daplinia magna)

2,183

0.0001

46,278

512.5
(285.3-920.4)

Bluegill (Lepomis macrocliirus)*

Fathead minnow (Pimephales
promelas)

1,938

0.0002

50,154

751.0
(390.0-1,446)

Bluegill (Lepomis macrocliirus)*

Mysid (Americamysis baliia)

372**

<0.0001

79.0

207.7
(107.0-403.2)

Brook trout (Salvelinus fontinalis)

Bluegill (Lepomis
macrocliirus)

6,452**

0.0004

123.3

3,505.7

(612.1-20,077)

Bullfrog (Lithobates catesbeianus)

Fathead minnow (Pimephales
promelas)

1,938

0.001

26,500

1,791.4

(756.1-4,244.6)

Bullfrog (Lithobates catesbeianus)

Bluegill (Lepomis
macrocliirus)

6,452**

0.0004

201.0

3,420.6°
(841.4-13,905)

Channel catfish (Ictalurus punctatus)

Fathead minnow (Pimephales
promelas)

1,938

0.001

26,500

1,617.3

(755.0-3,464.4)

Channel catfish (Ictalurus punctatus)

Bluegill (Lepomis
macrocliirus)

6,452**

0.0004

517.8

1,863.7°
(713.6-4,867.2)

Coho salmon (Oncorhynchus
kisutch)

Bluegill (Lepomis
macrocliirus)

6,452**

0.0004

150.8

1,609.5

(248.6-10,421)

Common carp (Cyprinus carpio)

Fathead minnow (Pimephales
promelas)

1,938**

0.005

133

1,952.5

(498.7-7,644.4)

Common carp (Cyprinus carpio)

Bluegill (Lepomis
macrocliirus)

6,452**

0.0009

180

2,962.4b
(569.2-15,418.0)

51


-------
Predicted Species

Surrogate

ICE Model
(m

Value Range

B/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Minimum

Maximum

Cutthroat trout (Oncorhynchus
clarkii)

Fathead minnow (Pimephales
promelas)

1,938**

0.001

140.2

602.6

(113.4-3,203.2)

Cutthroat trout (Oncorhynchus
clarkii)

Bluegill (Lepomis
macrochirus)

6,452**

0.0004

180

2,561.1

(548.4-11,961)

Daphnid (Ceriodaphnia dubia)

Mysid (Americamysis baliia)

372

<0.0001

4,100

171.7a
(36.6-804.9)

Daphnid (Ceriodaphnia dubia)

Daplinid (Daphnia magna)

2,183

0.0003

46,278

1,666.6

(909.0-3,055.6)

Daphnid (Daphnia magna)*

Mysid (Americamysis baliia)

372

<0.0001

11,000

243.9
(143.9-413.4)

Daphnid (Daphnia pulex)*

Daplinid (Daphnia magna)

2,183

0.0002

4,894.7

1,768.1
(893.9-3,497.1)

Fathead minnow (Pimephales
promelas)*

Mysid (Americamysis baliia)

372

<0.0001

11,000

158.6
(74.5-337.5)

Fathead minnow (Pimephales
promelas)*

Daplinid (Daphnia magna)

2,183

0.0002

46,500

639.5
(375.7-1,088.5)

Fathead minnow (Pimephales
promelas)*

Bluegill (Lepomis
macrochirus)

6,452

0.0001

27,540

3,399.2
(1,631.9-7080.3)

Fatmucket (Lampsilis siliquoidea)

Daplinid (Daphnia magna)

2,183

0.014

8,694.5

912.5

(399.0-2,086.8)

Goldfish (Carassius auratus)

Fathead minnow (Pimephales
promelas)

1,938

0.001

26,500

2,152.4

(1,075.2-4,309.0)

Goldfish (Carassius auratus)

Bluegill (Lepomis
macrochirus)

6,452**

0.0004

201

1,922.7 b
(312.9-11,813.0)

Green sunfish (Lepomis cyanellus)*

Bluegill (Lepomis
macrochirus)

6,452**

0.004

113

2,817.7
(563.5-14,089.5)

Guppy (Poecilia reticulata)

Fathead minnow (Pimephales
promelas)

1,938**

0.001

140.2

499.3

(131.8-1,892.4)

Guppy (Poecilia reticulata)

Bluegill (Lepomis
macrochirus)

6,452**

0.0004

180

687.3

(105.9-4,461.3)

Lake trout (Salvelinus namaycush)

Bluegill (Lepomis
macrochirus)

6,452**

0.003

180

320.8

(49.0-2,102.1)

Largemouth bass (Micropterus
salmoides)

Bluegill (Lepomis
macrochirus)

6,452**

0.0004

123.3

8,225.3

(2,938.4-23,024.8)

52


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Predicted Species

Surrogate

ICE Model
(m

Value Range

B/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Minimum

Maximum

Medaka (Oryzias latipes)

Fathead minnow (Pimephales
promelas)

1,938**

0.003

1,420

1,013.6

(496.2-2,070.7)

Midge (Paratanytarsus
parthenogeneticus)

Daphnid (Daplinia magna)

2,183

0.37

14,500

5,481.6

(1,674.6-17,943.7)

Neosho mucket (Lampsilis
rafinesqueana)

Daplinid (Daplinia magna)

2,183**

0.042

166.8

777.2

(115.1-5,248.1)

Nile tilapia (Oreochromis niloticus)

Bluegill (Lepomis
macrocliirus)

6,452**

0.002

6.969

11,873.0

(8,879.5-15,875.6)

Oriental river slirimp
(Macrobrachium nipponense)

Daplinid (Daplinia magna)

2,183**

0.011

281.6

2,722.2

(587.7-12,609.0)

Paper pondshell (Utterbackia
imbecillis)

Daplinid (Daplinia magna)

2,183

0.014

8,694.5

802.5

(422.2-1,525.5)

Rainbow trout (Oncorhynchus
mykiss)

Daplinid (Daplinia magna)

2,183

0.0001

14,500

361.9b
(221.2-592.1)

Rainbow trout (Oncorhynchus
mykiss)

Fathead minnow (Pimephales
promelas)

1,938

0.0002

26,500

788.9

(505.7-1,230.8)

Rainbow trout (Oncorhynchus
mykiss)

Bluegill (Lepomis
macrocliirus)

6,452

0.0001

7,100

3,208.6

(2,275.7-4,524.0)

Rainbow trout (Oncorhynchus
mykiss)

Mysid (Americamysis baliia)

372**

<0.0001

113

110.03
(58.8-205.9)

Redear sunfish (Lepomis
microlophus)*

Bluegill (Lepomis
macrocliirus)

6,452**

0.017

4.5

9,763.6
(7,469.9-12,761.7)

Swamp lymnaea (Lymnaea
stagnalis)

Daplinid (Daplinia magna)

2,183

0.014

8,694.5

1,523.7

(337.4-6,881.2)

Tadpole physa (Physella gyrina)

Daplinid (Daplinia magna)

2,183

0.014

8,694.5

1,608.6

(512.8-5,046.1)

Tlireeridge (Amblema plicata)

Fathead minnow (Pimephales
promelas)

1,938

0.814

10,831

250.3b
(39.4-1,592.2)

Tlireeridge (Amblema plicata)

Daplinid (Daplinia magna)

2,183

0.014

4,894.7

383.6

(85.6-1,717.6)

Vernal pool fairy slirimp
(Branchinecta lynclii)

Daplinid (Daplinia magna)

2,183

0.014

8,694.5

1,246.4

(457.0-3,399.4)

Washboard (Megalonaias nervosa)

Daplinid (Daplinia magna)

2,183

0.014

8,694.5

755.1

(345.4-1,650.5)

53


-------
Predicted Species

Surrogate

ICE Model
(m

Value Range

B/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Minimum

Maximum

Water flea (Chydorus sphaericus)

Daphnid (Daphnia magna)

2,183**

0.009

977.6

661.4

(349.2-1,252.8)

Western pearlshell (Margaritifera
falcata)

Daphnid (Daphnia magna)

2,183

0.014

8,694.5

706.1

(255.8-1,948.9)

Yellow perch (Perca flavescens)

Bluegill (Lepomis
macrochirus)

6,452**

0.0004

9.41

3,426.2

(710.8-16,515)

Zebrafish (Danio rerio)*

Fathead minnow (Pimephales
promelas)

1,938**

0.013

26.69

1,069.6
(242.1-4,726.4)

Zebrafish-embryo (Danio rerio -
embryo)*

Daphnid (Daphnia magna)

2,183

0.0001

46,500

780.8
(257.2-2,370.6)

Zebrafish-embryo (Danio rerio-
embryo)*

Fathead minnow (Pimephales
promelas)

1,938

0.004

70,200

1,707.9
(1,061.5-2,748)

* Acceptable models that were not used because genus level empirical data were available.

**Measured ECso falls outside range of ICE model and predicted SMAV is based on extrapolation beyond model bounds.
a Saltwater surrogate species were not used.

b Not used because other more closely taxonomically-related models were available.

0 Not used because other more closely taxonomically-related models where the measured surrogate ECso values fell within the ICE model range were available.

54


-------
Table 5-15. Ranked PFBS Genus Mean Acute Values.

Values in bold and highlighted are derived from empirical PFBS toxicity tests with the species.

Rank

MDR
Group1

Name

Species

SMAV
(mg/L)

GMAV
(mg/L)

Percentile
Rank

1

G

Threeridge

Amblema plicata

383.6

383.6

0.03

2

E

Amphipod

Gammctrus fascia tus

428.1

428.1

0.06

3

B

Guppy

Poecilia reticulata

585.8**

585.8

0.10

4

D

Water flea

Chydorus sphaericus

661.4**

661.4

0.13

5

G

Western pearlshell

Margaritifera falcata

706.1

706.1

0.16

6

G

Washboard

Megalonaias nervosa

755.1

755.1

0.19

7

G

Paper pondshell

Utterbackia imbecillis

802.5

802.5

0.23

8

G

Neosho mucket

Lampsilis raflnesqueana

777.2**

842.1

0.26

G

Fatmucket

Lampsilis siliquoidea

912.5

9

E

Beaver-tail fairy
shrimp

Thamnocephalus
platyurus

1,010

1,010

0.29

10

B

Medaka

Oryzias latipes

1,014**

1,014

0.32

11

A

Brook trout

Salve linns fontinalis

3,506**

1,060

0.35

A

Lake trout

Salvelinus namavcush

320.8**

12

E

Vernal pool fairy
shrimp

Branchinecta lvnchi

1,246

1,246

0.39

13

A

Cutthroat trout

Oncorhvnchiis clarkii

1,242**

1,471

0.42

A

Coho salmon

Oncorhvnchiis kisutch

1,610**

A

Rainbow trout

Oncorhvnchiis mykiss

1,591

14

G

Swamp lymnaea

Lymnaea stagnalis

1,524

1,524

0.45

15

G

Tadpole physa

Physella gyrina

1,609

1,609

0.48

16

B

Channel catfish

Ictalurus punctatus

1,617

1,617

0.52

17

D

Daphnid

Ceriodaphnia dubia

1,667

1,667

0.55

18

C

Bullfrog

Lithobates catesbeianus

1,791

1,791

0.58

19

B

Fathead minnow

Pimephales promelas

1,938

1,938

0.61

20

B

Common carp

Cypriniis carpio

1,952**

1,952

0.65

21

B

Goldfish

Carassins auratus

2,152

2,152

0.68

22

D

Daphnid

Daphnia magna

2,183

2,183

0.71

23

E

Oriental river shrimp

Macrobrachium
nipponense

2,722**

2,722

0.74

24

B

Zebrafish

Danio rerio

>3,000

>3,000

0.77

25

B

Yellow perch

Percaflavescens

3,426**

3,426

0.81

26

F

Midge

Paratanv tarsus
parthenogeneticus

5,482

5,482

0.84

27

B

Bluegill

Lepomis macrochirus

6,452

6,452

0.87

28

B

Largemouth bass

Micropterus salmoides

8,225**

8,225

0.90

29

B

Nile tilapia

Oreochromis niloticus

11,873**

11,873

0.94

30

A

Atlantic salmon

Salmo salar

14,524**

14,524

0.97

55


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**Measured EC50 falls outside range of ICE model and predicted SMAV is based on extrapolation beyond model

bounds.

1: Freshwater MDR Groups

A)	The family Salmonidae in the class Osteichthyes

B)	A second family in the class Osteichthyes, preferably a commercially or recreationally important

warmwater species (e.g., bluegill, channel catfish, etc.)

C)	A third family in the phylum Chordata (may be in the class Osteichthyes or may be an amphibian, etc.)

D)	A planktonic crustacean (e.g., cladoceran, copepod, etc.)

E)	Abenthic crustacean (e.g., ostracod, isopod, amphipod, crayfish, etc.)

F)	An insect (e.g., mayfly, dragonfly, damselfly, stonefly, caddisfly, mosquito, midge, etc.)

G)	A family in a phylum other than Arthropoda or Chordata (e.g., Rotifera, Annelida, Mollusca, etc.)

H)	A family in any order of insect or any phylum not already represented.

Table 5-16. PFBS Protective Aquatic Life Acute Benchmark.

Calculated Freshwater FAY based on 4 lowest values; N=30 GMAVs total





Benchmark calculated by dividing the FAY by 2 and by the sulfonic acid application factor (42.3)





GMAV









Rank

Genus

(mg/L)

ln(GMAV)

ln(GMAV)2

P=R/(N+1)

sqrt(P)

1

Amblema

383.6

5.95

35.40

0.032

0.180

2

Gammarus

428.1

6.06

36.72

0.065

0.254

3

Hyalella

585.8

6.37

40.61

0.097

0.311

4

Ceriodaphnia

661.4

6.49

42.18

0.129

0.359



£ (Sum):

24.88

154.90

0.32

1.10







P = cumulative probability











R = rank













N = number of GMAVs





S2 =

11.02



S = slope







L =

5.303



L = X-axis intercept





A =

6.045



A = InFAV







FAV =

422.1











FAV/2 =

211.0 mg/L (Preliminary Value)







Adjustment =

211.0 / 42.3 = 4.989 (Preliminary Value / Sulfonic Acid Application Factor)

Benchmark =

5.0 mg/L PFBS (rounded to two significant figures)





56


-------
1.00 -T
0.90

a 0.80 -¦
o

w

C3

0.70
0.60

u

X 0.50 -¦
s

C5

es

£ 0.40 4
a

2

t 0.30 4

0.20
0.10
0.00

1

~

o
+
~

A

O

Amphibian (WeblCE)
Fish (Empirical)

Fish (WeblCE)

Insect (WeblCE)
Invertebrate (Empirical)
Invertebrate (WeblCE)
Mollusk (WeblCE)
Preliminary V alue
¦ Acute Benchmark

O Salmo
O Oreochromis
O Micropterus
# Lepomis
+ Paratanvtarsus
O Perca
# Danio

A Macrobrachium
A Daphnia

O Carassius
O Cyprinus
# Pimephales
~ Lithobates
A Ceriodaphnia
O Ictalurus
O Physella
O Lymnaea
O Oncorhvnchus
A Branchinecta
O Salvelinus
O Oryzias
A Thamnocephalus
O Lampsilis
O Utterbackia
O Megalonaias
<0 Margaritifera
A Chydorus
O Poecilia
A Gammarus
O Amblema

10	100	1,000

Genus Mean Acute Value (mg/L PFBS)

10,000

100,000

Figure 5-5. Ranked Acute PFBS GMAVs Used for the Aquatic Life Acute Benchmark
Calculation.

5.7 Derivation of Acute Water Benchmark for Perfluorohexanesulfonic
Acid (PFHxS)

Quantitatively acceptable empirical data for PFHxS were available for three species
comprising two genera and fulfilling two MDR groups (Tables 4-2 and 5-18). Because only two
of the MDRs were met with empirical data, ICE-derived values were used to fulfill the remaining
six MDRs. A total of 17 ICE models were acceptable for use. Of these, four models were not
used because there were acceptable models for more closely-related surrogate species (i.e.,
Lithobates catesbeicmus predicting for Ictaluruspunctatus\ Lithobates catesbeianus predicting

57


-------
for Pimephalespromelas; Lithobates catesbeianus predicting for Carassius auratus; and
Lithobates catesbeianus predicting for Oncorhynchus mykiss). A total of 13 ICE models were
selected for use, resulting in ICE models predicting 10 SMAVs representing 10 genera and five
MDRs (Tables 5-17 and 5- 18). ICE models were not available to fulfill two MDRs: an insect,
and a family in a phylum other than Arthropoda or Chordata.

The combined empirical and ICE data resulted in 12 GMAVs that collectively fulfill six
of the eight MDR groups for deriving a freshwater criterion as defined by the Aquatic Life
Criteria Guidelines (U.S. EPA 1985). The ranked GMAVs for these combined data along with
the MDR met by each GMAV are summarized in Table 5-18. GMAVs for the four most
sensitive genera were within a factor of 1.2 of each other (Table 5-19). The freshwater FAV (the
5th percentile of the genus sensitivity distribution) for PFHxS is 18.07 mg/L (Table 5-19), which
is lower than all of the GMAVs. The FAV was divided by two to obtain a preliminary value of
9.035 mg/L PFHxS and then adjusted by the sulfonic acid application factor (42.3) to obtain the
freshwater acute water column benchmark magnitude of 0.21 mg/L PFHxS (rounded to two
significant figures). This value is expected to be protective of 95% of freshwater genera exposed
to PFHxS under short-term conditions of one-hour duration, if the one-hour average magnitude is
not exceeded more than once in three years (Figure 5-6). However, because only six of the eight
MDRs were fulfilled for PFHxS (when both empirical test and ICE data were considered), there
is considered to be greater uncertainty associated with this benchmark value compared with other
PFAS benchmarks developed.

58


-------
Table 5-17. Acceptable Models for ICE-estimated Species Sensitivity to PFHxS.

Includes values that were extrapolated as indicated. Bold predicted EC50S used for SMAY calculations.

Predicted Species

Surrogate

ICE Model
(m

Value Range
2/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Minimum

Maximum

Bluegill (Lepomis macrochirus)

Bullfrog (Lithobates
catesbeianus)

1,105**

0.003

233.0

1,576.9

(395.3-6,289.9)

Channel catfish (Ictalurus punctatus)

Zebrafish-embryo (Danio rerio-
embryo)

22.5

0.145

304.5

19.5

(9.2-41.0)

Channel catfish (Ictalurus punctatus)

Bullfrog (Lithobates
catesbeianus)

1,105

0.003

20,900

719.8b
(362.4-1,429.7)

Daphnid (Daphnia magna)

Zebrafish (Danio rerio)

22.5

0.003

36.91

5.2

(1.2-22.7)

Daphnid (Daphnia magna)

Bullfrog (Lithobates
catesbeianus)

1,105

0.003

20,900

424.1

(68.5-2,627.9)

Fairy shrimp (Streptocephalus
rubricaudatus)

Zebrafish-embryo (Danio rerio-
embryo)

22.5

4.622

5,100.7

114.2

(33.4-390.2)

Fathead minnow (Pimephales
promelas)

Zebrafish-embryo (Danio rerio-
embryo)

22.5

0.023

54,579

18.3

(13.3-25.2)

Fathead minnow (Pimephales
promelas)

Zebrafish (Danio rerio)

22.5

0.018

25.08

26.0

(11.6-58.2)

Fathead minnow (Pimephales
promelas)

Bullfrog (Lithobates
catesbeianus)

1,105

0.003

20,900

1,083.6b
(468.5-2,506.4)

Flagfish (Jordanella floridae)

Zebrafish (Danio rerio)

22.5

7.797

649.3

33.9

(19.7-58.4)

Goldfish (Carassius auratus)

Zebrafish-embryo (Danio rerio-
embryo)

22.5

0.145

304.5

37.3

(23.2-59.9)

Goldfish (Carassius auratus)

Bullfrog (Lithobates
catesbeianus)

1,105

0.003

20,900

1,027.7b
(405.9-2,602.1)

Mosquitofish (Gambusia affinis)

Bullfrog (Lithobates
catesbeianus)

1,105

18.6

20,900

805.1

(394.5-1,643.4)

Oligochaete (Limnodrilus
hoffmeisteri)

Zebrafish-embryo (Danio rerio-
embryo)

22.5

0.145

239.3

63.2

(9.9-403.1)

Rainbow trout (Oncorhynchus
mykiss)

Zebrafish-embryo (Danio rerio-
embryo)

22.5

0.023

8,843.9

9.6

(4.2-22.0)

59


-------
Predicted Species

Surrogate

ICE Model
(m

Value Range
2/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Minimum

Maximum

Rainbow trout (Oncorhynchus
mykiss)

Zebrafish (Danio rerio)

22.5

0.003

26.39

38.3

(9.5-155.1)

Rainbow trout (Oncorhynchus
mykiss)

Bullfrog (Lithobates
catesbeianus)

1,105

0.003

20,900

713.5b
(200.3-2,541.4)

* *Measured ECso falls outside range of ICE model and predicted SMAV is based on extrapolation beyond model bounds.
a Saltwater surrogate species were not used.

b Not used because other more closely taxonomically-related models were available.

0 Not used because other more closely taxonomically-related models where the measured surrogate ECso values fell within the ICE model range were available.

60


-------
Table 5-18. Ranked PFHxS Genus Mean Acute Values.

Values in bold and highlighted are derived from empirical PFHxS toxicity tests with the species.



MDR





SMAV

GMAV

Percentile

Rank

Group1

Name

Species

(mg/L)

(mg/L)

Rank

1

A

Rainbow trout

Oncorhvnchus mvkiss

19.19

19.19

0.08

2

B

Channel catfish

Ictcdurus punctatiis

19.46

19.46

0.15

3

B

Fathead minnow

Pimephales promelas

21.79

21.79

0.23

4

B

Zebrafish

Danio rerio

22.50

22.50

0.31

5

B

Flagfish

Jordanellct floridae

33.90

33.90

0.38

6

B

Goldfish

Cctrctssius auratus

37.28

37.28

0.46

7

D

Daphnid

Daphnia magna

46.87

46.87

0.54

8

H

Oligochaete

Limnodrilus hoffmeisteri

63.23

63.23

0.62

9

E

Fairy shrimp

Streptocephalus
nibricaiidatiis

114.2

114.2

0.69

10

B

Mosquitofish

Gambusia afftnis

805.1

805.1

0.77

11

C

Bullfrog

Lithobates catesbeiana

1,105

915.2

0.85

C

Green frog

Lithobates clamitans

758.0

12

B

Bluegill

Lepomis macrochirns

1,577**

1,577

0.92

**Measured EC50 falls outside range of ICE model and predicted SMAV is based on extrapolation beyond model

bounds.

1: Freshwater MDR Groups

A)	The family Salmonidae in the class Osteichthyes

B)	A second family in the class Osteichthyes, preferably a commercially or recreationally important

warmwater species (e.g., bluegill, channel catfish, etc.)

C)	A third family in the phylum Chordata (may be in the class Osteichthyes or may be an amphibian, etc.)

D)	A planktonic crustacean (e.g., cladoceran, copepod, etc.)

E)	Abenthic crustacean (e.g., ostracod, isopod, amphipod, crayfish, etc.)

F)	An insect (e.g., mayfly, dragonfly, damselfly, stonefly, caddisfly, mosquito, midge, etc.)

G)	A family in a phylum other than Arthropoda or Chordata (e.g., Rotifera, Annelida, Mollusca, etc.)

H)	A family in any order of insect or any phylum not already represented.

61


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Table 5-19. PFHxS Final Acute Value and Protective Aquatic Acute Benchmark.

Bold values represent genera for which empirical toxicity data were available.	

Calculated Freshwater FAY based on 4 lowest values; N=12 GMAVs total





Benchmark calculated by dividing the FAY by 2 and by the

sulfonic acid application factor (42.3)





GMAV









Rank

Genus

(mg/L)

ln(GMAV)

ln(GMAV)2

P=R/(N+1)

sqrt(P)

1

Oncorhynchus

19.19

2.95

8.73

0.077

0.277

2

Ictalurus

19.46

2.97

8.81

0.154

0.392

3

Pimephales

21.79

3.08

9.50

0.231

0.480

4

Danio

22.50

3.11

9.69

0.308

0.555



£ (Sum):

12.12

36.73

0.77

1.70







P = cumulative probability











R = rank













N = number of GMAVs





S2 =

0.45



S = slope







L =

2.745



L = X-axis intercept





A =

2.894



A = InFAV







FAV =

18.07











FAV/2 =

9.035 mg/L (Preliminary Value)







Adjustment =

9.035 / 42.3 = 0.2136 (Preliminary Value / Sulfonic Acid Application Factor)

Benchmark =

0.21 mg/L PFHxS (rounded to two significant figures)





62


-------


E

9

1.00	-T

0.90	-J

0.80	-•

0.70	-¦
0.60

JZ 0.50 -
a

R

06

0.40 -¦


-------
acceptable models for more closely-related surrogate species (i.e., Daphnia magna predicting for
Lepomis macrochirus; Daphnia magna predicting for Pimephalespromelas; Daphnia magna
predicting for Danio rerio). A total of 53 ICE models were selected for use, resulting in ICE
models predicting 52 SMAVs representing 43 genera and all eight MDRs (Tables 5-20 and 5-
21).

The combined empirical and ICE data resulted in 45 GMAVs that collectively fulfill the
eight MDR groups for deriving a freshwater criterion as defined by the Aquatic Life Criteria
Guidelines (U.S. EPA 1985). The ranked GMAVs for these combined data along with the MDR
met by each GMAV is summarized in Table 5-21. GMAVs for the four most sensitive genera
were within a factor of 1.8 of each other (Table 5-22). The freshwater FAV (the 5th percentile of
the genus sensitivity distribution) for 8:2 FTUCA is 1.171 mg/L (Table 5-22), which is lower
than all of the GMAVs except for the ICE-derived species value for the threeridge mussel,
Amblemaplicata (GMAV=0.79 mg/L). The FAV was divided by two to obtain a preliminary
value of 0.5853 mg/L 8:2 FTUCA and then adjusted by the carboxylic acid application factor
(15.8) to obtain the freshwater acute water column benchmark magnitude of 0.037 mg/L 8:2
FTUCA (rounded to two significant figures). This value is expected to be protective of 95% of
freshwater genera exposed to 8:2 FTUCA under short-term conditions of one-hour duration, if
the one-hour average magnitude is not exceeded more than once in three years (Figure 5-7).

There is a visible gap in the data between the water flea (Miona macrocopa, SMAV = 4.2
mg/L) and bryozoan (Pectinatella magnifica, SMAV =15.2 mg/L) for these ICE-generated
values (see Figure 5-7). Although the exact reason is unclear for this large gap in values, it is
noted that the lower portion of the curve is primarily composed of the phyla Mollusca and
Arthropoda, while the upper portion of the curve is primarily composed of the phylum Chordata.

64


-------
The distinct difference in sensitivity indicated by the gap in these data is thus likely to result
from a greater estimated sensitivity of most invertebrates than vertebrates to 8:2 FTUCA.
Additionally, the ICE estimates for the lower portion of the curve are based primarily on the
surrogate Daphnia magna, while the ICE estimates for the upper portion of the curve are based
primarily on the surrogate Oncorhynchns mykiss. However, these differences fall within the
range of differences in acute response frequently observed between species. For example, within
the PFOA and PFOS acute criteria datasets, there was single species sensitivity that spanned over
two orders of magnitude, and the variation in acute toxicity values is frequently an order of
magnitude and can be as high as three orders of magnitude (Hrovat et al. 2009, Schiir et al. 2023,
Raimondo et al. 2024). It is, therefore, not unexpected to see the difference in sensitivity
observed between these species.

65


-------
Table 5-20. Acceptable Models for ICE-estimated Species Sensitivity to 8:2 FTUCA.

Includes values that were extrapolated as indicated. Bold predicted EC50S used for SMAY calculations.

Predicted Species

Surrogate

ICE Model
(m

Value Range
2/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Minimum

Maximum

Amphipod (Crangonyx
pseudogracilis)

Daphnid (Daplinia magna)

3.2

0.0008

166.8

28.6

(7.0-116.1)

Amphipod (Gammarus fasciatus)

Daplinid (Daplinia magna)

3.2

0.0002

5,000

2.0

(1.0-4.1)

Amphipod (Gammarus
pseudolimnaeus)

Daphnid (Daplinia magna)

3.2

0.0001

68.3

2.9

(1.0-7.8)

Apache trout (Oncorhynchus gilae)*

Rainbow trout (Oncorhynchus
mykiss)

81**

0.004

1.625

109.4
(20.3-589.1)

Atlantic salmon (Salmo salar)

Rainbow trout (Oncorhynchus
mykiss)

81

0.0002

95.86

79.3

(36.6-171.6)

Beaver-tail fairy shrimp
(Thamnocephalus platyurus)

Daplinid (Daplinia magna)

3.2

0.0003

8,694.5

2.6

(1.8-3.9)

Bluegill (Lepomis macrocliirus)

Daplinid (Daplinia magna)

3.2

0.0001

46,278

4.9b
(3.7-6.4)

Bluegill (Lepomis macrocliirus)

Rainbow trout (Oncorhynchus
mykiss)

81

0.0001

8,341.5

79.0

(64.3-97.1)

Brook trout (Salvelinus fontinalis)

Rainbow trout (Oncorhynchus
mykiss)

81

0.0006

96.69

79.4

(44.1-142.9)

Brown trout (Salmo trutta)

Rainbow trout (Oncorhynchus
mykiss)

81

0.0002

95.86

83.8

(47.0-149.4)

Bryozoan (Pectinatella magnifica)

Daplinid (Daplinia magna)

3 2**

0.014

0.232

15.2

(3.5-66.6)

Bullfrog (Lithobates catesbeianus)

Rainbow trout (Oncorhynchus
mykiss)

81

0.0006

13,400

132.5

(51.0-344.2)

Cape Fear shiner (Notropis
mekistocholas)

Rainbow trout (Oncorhynchus
mykiss)

81**

0.004

1.625

339.1

(82.8-1,388.7)

Channel catfish (Ictalurus punctatus)

Rainbow trout (Oncorhynchus
mykiss)

81

<0.0001

13,400

67.4

(39.8-114.0)

Cliinook salmon (Oncorhynchus
tshawytscha)*

Rainbow trout (Oncorhynchus
mykiss)

81

0.003

724.4

102.9
(57.1-185.4)

66


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Predicted Species

Surrogate

ICE Model
(m

Value Range
2/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Minimum

Maximum

Coho salmon (Oncorhynchus
kisutch)*

Rainbow trout (Oncorhynchus
mykiss)

81**

<0.0001

43.07

128.3
(96.6-170.5)

Common carp (Cyprinus carpio)

Rainbow trout (Oncorhynchus
mykiss)

81

0.0002

198.4

75.2

(31.0-182.2)

Cuttliroat trout (Oncorhynchus
clarkii)*

Rainbow trout (Oncorhynchus
mykiss)

81

<0.0001

198.4

65.5
(42.9-100.0)

Cuvier's foam froglet (Physalaemus
cuvieri)

Rainbow trout (Oncorhynchus
mykiss)

81

0.7099

155.5

91.1

(48.4-171.5)

Daphnid (Ceriodaplinia dubia)

Daplinid (Daplinia magna)

3.2

0.0003

46,278

2.1

(1.5-2.9)

Daplinid (Daplinia galeata)*

Daplinid (Daplinia magna)

3 2**

0.0001

0.646

3.1
(0.8-12.3)

Daplinid (Daplinia longispina)*

Daplinid (Daplinia magna)

3.2

0.009

10.36

4.9
(1.0-23.7)

Daplinid (Daplinia pulex)*

Daplinid (Daplinia magna)

3.2

0.0002

4,894.7

2.4
(1.7-3.3)

Daplinid (Daplinia pulicaria)*

Daplinid (Daplinia magna)

3.2

0.014

281.6

2.6
(0.7-9.5)

Daplinid (Simocephalus serrulatus)

Daplinid (Daplinia magna)

3.2

0.0002

7.2

1.8

(0.7-5.2)

Daplinid (Simocephalus vetulus)

Daplinid (Daplinia magna)

3.2

0.0001

166.8

1.7

(0.2-12.3)

Fathead minnow (Pimephales
promelas)

Daplinid (Daplinia magna)

3.2

0.0002

46,500

7.0b
(5.4-9.0)

Fathead minnow (Pimephales
promelas)

Rainbow trout (Oncorhynchus
mykiss)

81

0.0002

13,400

107.7

(79.8-145.3)

Fatmucket (Lampsilis siliquoidea)

Daplinid (Daplinia magna)

3.2

0.014

8,694.5

2.5

(1.6-4.1)

Goldfish (Carassius auratus)

Rainbow trout (Oncorhynchus
mykiss)

81

<0.0001

13,400

132.7

(62.2-283.3)

Green floater (Lasmigona subviridis)

Daplinid (Daplinia magna)

3.2

0.014

166.8

1.9

(0.4-9.4)

Green sunfish (Lepomis cyanellus)

Rainbow trout (Oncorhynchus
mykiss)

81**

<0.0001

19.93

105.8

(41.7-269)

67


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Predicted Species

Surrogate

ICE Model
(m

Value Range
2/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Minimum

Maximum

Guppy (Poecilia reticulata)

Rainbow trout (Oncorhynchus
mykiss)

81

0.0006

198.4

54.9

(12.3-243.7)

Isopod (Asellus aquaticus)

Daphnid (Daplinia magna)

3.2

0.0003

166.8

76.6

(23.0-255.1)

Isopod (Caecidotea brevicauda)

Rainbow trout (Oncorhynchus
mykiss)

81

0.0006

82

16.8

(3.2-89.7)

Lake trout (Salvelinus namaycush)

Rainbow trout (Oncorhynchus
mykiss)

81

0.0002

198.4

41.3

(24.5-69.7)

Largemouth bass (Micropterus
salmoides)

Rainbow trout (Oncorhynchus
mykiss)

81

<0.0001

95.86

46.8

(20.6-106.2)

Medaka (Oryzias latipes)

Rainbow trout (Oncorhynchus
mykiss)

81**

0.007

54.4

121.6

(20.2-733.2)

Midge (Paratanytarsus dissimilis)

Rainbow trout (Oncorhynchus
mykiss)

81

0.0006

1,330

241.0

(72.2-804.7)

Midge (Paratanytarsus
parthenogeneticus)

Daphnid (Daplinia magna)

3.2

0.37

14,500

9.3

(3.5-24.7)

Mississippi grass shrimp
(Palaemonetes kadiakensis)

Daphnid (Daplinia magna)

3.2

0.0003

58

1.3

(0.3-6.2)

Neosho mucket (Lampsilis
rafinesqueana)

Daphnid (Daplinia magna)

3.2

0.042

166.8

2.3

(1.1-5.1)

Northern leopard frog (Lithobates
pipiens)

Rainbow trout (Oncorhynchus
mykiss)

81

0.002

198.4

125.5

(62.3-252.8)

Oligochaete (Limnodrilus
hoffmeisteri)

Daphnid (Daplinia magna)

3.2

0.003

281.6

17.9

(6.9-46.2)

Oligochaete (Tubifex tubifex)

Daphnid (Daplinia magna)

3.2

0.0001

4,894.7

31.6

(7.4-134.7)

Oriental river shrimp (Macrobrachium
nipponense)

Daphnid (Daplinia magna)

3.2

0.011

281.6

1.5

(0.7-3.4)

Paper pondshell (Utterbackia
imbecillis)

Daphnid (Daplinia magna)

3.2

0.014

8,694.5

2.5

(1.6-3.7)

Pheasantshell (Ortmanniana
pectorosa)

Daphnid (Daplinia magna)

3.2

0.042

545.9

2.3

(0.8-7.0)

Rainbow trout (Oncorhynchus
my kiss)*

Daphnid (Daplinia magna)

3.2

0.0001

14,500

3.4
(2.7-4.3)

68


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Predicted Species

Surrogate

ICE Model
(m

Value Range
2/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Minimum

Maximum

Razorback sucker (Xyrauchen
texanus)

Rainbow trout (Oncorhynchus
mykiss)

81**

0.003

43.07

54.8

(8.5-352.2)

Shortnose sturgeon (Acipenser
brevirostrum)

Rainbow trout (Oncorhynchus
mykiss)

81

0.028

95.86

134.5

(26.7-677.1)

Silver perch (Bidyanus bidyanus)

Rainbow trout (Oncorhynchus
mykiss)

81**

0.0014

7.075

200.3

(73.6-544.8)

Southern leopard frog (Lithobates
sphenocephalus)

Rainbow trout (Oncorhynchus
mykiss)

81**

0.028

9.7

1,023.9

(231.9-4,520.4)

Swamp lymnaea (Lyinnaea stagnalis)

Daphnid (Daphnia magna)

3.2

0.014

8,694.5

3.2

(1.2-8.4)

Tadpole physa (Physella gyrina)

Daphnid (Daplinia magna)

3.2

0.014

8,694.5

3.0

(1.4-6.7)

Tlireeridge (Amblema plicata)

Daplinid (Daplinia magna)

3.2

0.014

4,894.7

0.8

(0.3-2.1)

Vernal pool fairy shrimp
(Branchinecta lynchi)

Daphnid (Daplinia magna)

3.2

0.014

8,694.5

2.6

(1.4-4.9)

Washboard (Megalonaias nervosa)

Daplinid (Daplinia magna)

3.2

0.014

8,694.5

1.4

(0.8-2.6)

Water flea (Chydorus sphaericus)

Daplinid (Daplinia magna)

3.2

0.009

977.6

1.3

(0.8-2.2)

Water flea (Moina macrocopa)

Daplinid (Daplinia magna)

3.2

0.009

281.6

4.2

(1.3-13.2)

Western pearlshell (Margaritifera
falcata)

Daplinid (Daplinia magna)

3.2

0.014

8,694.5

2.3

(1.1-4.6)

Yellow perch (Perca flavescens)

Rainbow trout (Oncorhynchus
mykiss)

81**

<0.0001

16.24

51.2

(10.0-262.6)

Zebrafish (Danio rerio)

Daplinid (Daplinia magna)

3.2

0.0001

50

8.3b
(2.5-28.0)

Zebrafish (Danio rerio)

Rainbow trout (Oncorhynchus
mykiss)

81

0.0002

229.1

34.6

(11.3-106.0)

Zebrafish-embryo (Danio rerio-
embryo)

Daplinid (Daplinia magna)

3.2

0.0001

46,500

10.6b
(5.7-19.9)

Zebrafish-embryo (Danio rerio-
embryo)

Rainbow trout (Oncorhynchus
mykiss)

81

0.0002

8,341.5

65.2

(29.0-146.3)

69


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*Acceptable models that were not used because genus level empirical data were available.

* *Measured EC50 falls outside range of ICE model and predicted SMAV is based on extrapolation beyond model bounds.
a Saltwater surrogate species were not used.

b Not used because other more closely taxonomically-related models were available.

0 Not used because other more closely taxonomically-related models where the measured surrogate EC50 values fell within the ICE model range were available.

70


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Table 5-21. Ranked 8:2 FTUCA Genus Mean Acute Values.

Rank

MDR
Group1

Name

Species

SMAV
(mg/L)

GMAV
(mg/L)

Percentile
Rank

1

G

Threeridge

Amblema plicata

0.7894

0.7894

0.02

2

E

Mississippi grass
shrimp

Palaemonetes
kadictkensis

1.327

1.327

0.04

3

D

Water flea

Chydorus sphaericus

1.344

1.344

0.07

4

G

Washboard

Megalonaias nervosa

1.403

1.403

0.09

5

E

Oriental river shrimp

Macrobrachium
nipponense

1.518

1.518

0.11

6

D

Daphnid

Simocephalus serrulatus

1.848

1.795

0.13

D

Daphnid

Simocephalus vetulus

1.743

7

G

Green floater

Lasmigona subviridis

1.900

1.900

0.15

8

D

Daphnid

Ceriodaphnia dubia

2.066

2.066

0.17

9

G

Western pearlshell

Margaritifera falcata

2.268

2.268

0.20

10

G

Pheasantshell

Ortmanniana pectorosa

2.347

2.347

0.22

11

E

Amphipod

Gammctrus fascia tus

2.040

2.412

0.24

E

Amphipod

Gammarus
pseiidolimnaens

2.853

12

G

Neosho mucket

Lampsilis rafinesqueana

2.325

2.430

0.26

G

Fatmucket

Lampsilis siliquoidea

2.540

13

G

Paper pondshell

Utterbackia imbecillis

2.454

2.454

0.28

14

E

Beaver-tail fairy
shrimp

Thamnocephalus
platyurus

2.632

2.632

0.30

15

E

Vernal pool fairy
shrimp

Branchinecta lvnchi

2.645

2.645

0.33

16

G

Tadpole physa

Physella gyrina

3.035

3.035

0.35

17

D

Daphnid

Daphnia magna

3.200

3.200

0.37

18

G

Swamp lymnaea

Lymnaea stagnalis

3.210

3.210

0.39

19

D

Water flea

Moina macrocopa

4.192

4.192

0.41

20

H

Bryozoan

Pectinatella magnifica

15.21**

15.21

0.43

21

E

Isopod

Caecidotea brevicauda

16.82

16.82

0.46

22

H

Oligochaete

Limnodrilus hoffmeisteri

17.89

17.89

0.48

23

E

Amphipod

Crangonyx
pseudogracilis

28.57

28.57

0.50

24

H

Oligochaete

Tubifex tubifex

31.57

31.57

0.52

25

B

Largemouth bass

Micropterns salmoides

46.83

46.83

0.54

26

B

Zebraflsh

Danio rerio

47.47

47.47

0.57

27

F

Midge

Paratanv tarsus
dissimilis

241.0

47.45

0.59

F

Midge

P aratanv tarsus
parthenogeneticus

9.343

28

B

Yellow perch

Perca flavescens

51.18**

51.18

0.61

29

B

Razorback sucker

Xyrauchen texanus

54.83**

54.83

0.63

i the species.

71


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Rank

MDR
Group1

Name

Species

SMAV
(mg/L)

GMAV
(mg/L)

Percentile
Rank

30

B

Guppy

Poecilia reticulata

54.85

54.85

0.65

31

A

Brook trout

Salve linns fontinalis

79.36

57.25

0.67

A

Lake trout

Salvelinus namavcush

41.30

32

B

Channel catfish

Ictalurus punctatiis

67.38

67.38

0.70

33

B

Common carp

Cyprinus carpio

75.18

75.18

0.72

34

E

Isopod

Asellus aquaticus

76.56

76.56

0.74

35

A

Rainbow trout

Oncorhynchus mykiss

81.00

81.00

0.76

36

A

Atlantic salmon

Salmo salar

79.29

81.49

0.78

A

Brown trout

Salmo trutta

83.76

37

C

Cuvier's foam froglet

Phvsalaemus cuvieri

91.14

91.14

0.80

38

B

Green sunfish

Lepomis cyanellus

105.8**

91.43

0.83

B

Bluegill

Lepomis macrochirns

78.99

39

B

Fathead minnow

Pimephales promelas

107.7

107.7

0.85

40

B

Medaka

Oryzias latipes

121.6**

121.6

0.87

41

B

Goldfish

Carassius auratus

132.7

132.7

0.89

42

B

Shortnose sturgeon

Acipenser brevirostrum

134.5

134.5

0.91

43

B

Silver perch

Bidvanus bidvanus

200.3**

200.3

0.93

44

C

Bullfrog

Lithobates catesbeianus

132.5

257.2

0.96

C

Northern leopard frog

Lithobates pipiens

125.5

C

Southern leopard frog

Lithobates
sphenocephalns

1,024**

45

B

Cape Fear shiner

Notropis mekistocholas

339.1**

339.1

0.98

**Measured EC50 falls outside range of ICE model and predicted SMAV is based on extrapolation beyond model

bounds.

1: Freshwater MDR Groups

A)	The family Salmonidae in the class Osteichthyes

B)	A second family in the class Osteichthyes, preferably a commercially or recreationally important

warmwater species (e.g., bluegill, channel catfish, etc.)

C)	A third family in the phylum Chordata (may be in the class Osteichthyes or may be an amphibian, etc.)

D)	A planktonic crustacean (e.g., cladoceran, copepod, etc.)

E)	Abenthic crustacean (e.g., ostracod, isopod, amphipod, crayfish, etc.)

F)	An insect (e.g., mayfly, dragonfly, damselfly, stonefly, caddisfly, mosquito, midge, etc.)

G)	A family in a phylum other than Arthropoda or Chordata (e.g., Rotifera, Annelida, Mollusca, etc.)

H)	A family in any order of insect or any phylum not already represented.

72


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Table 5-22. 8:2 FTUCA Final Acute Value and Protective Aquatic Acute Benchmark.

Calculated Freshwater FAY based on 4 lowest values; N=45 GMAVs total





Benchmark calculated by dividing the FAY by 2 and by the carboxylic acid application factor (15.8)





GMAV









Rank

Genus

(mg/L)

ln(GMAV)

ln(GMAV)2

P=R/(N+1)

sqrt(P)

1

Amblema

0.7894

-0.24

0.06

0.022

0.147

2

Palaemonetes

1.327

0.28

0.08

0.043

0.209

3

Chydorus

1.344

0.30

0.09

0.065

0.255

4

Megalonaias

1.403

0.34

0.11

0.087

0.295



£ (Sum):

0.68

0.34

0.22

0.91







P = cumulative probability











R = rank













N = number of GMAVs





S2 =

18.39



S = slope







L =

-0.801



L = X-axis intercept





A =

0.158



A = InFAV







FAV =

1.171











FAV/2 =

0.5853 mg/L (Preliminary Value)







Adjustment =

0.5853 / 15.8 = 0.03705 (Preliminary Value / Carboxylic Acid Application Factor)

Benchmark =

0.037 mg/L 8:2 FTUCA (rounded to two significant figures)



73


-------
1.00 T
0.90 ¦ -
0.80



C3 0.70 --



•3 0.60 »

s

3

0.50

~

Amphibian (WeblCE)

•

Fish (Empirical)

o

Fish (WeblCE)

+

Insect (WeblCE)

A

Invertebrate (Empirical)

A

Invertebrate (WeblCE)

©

Mollusk (WeblCE)



• Preliminary Value



-Acute Benchmark

A Moina
O Lymnaea
~ D'aphnia
O Physella
A Branchinecta
A Thamnocephalus
O Utterbackia
O Lampsilis
A Gammarus
O Ortmanniana
O Margaritifera
A Ceriodaphnia
O Lasmigona
A Simocephalus
A Macrobrachium
O Megalonaias
A Chvoorus
A Palaemonetes
Amblema

O Notropis
~ Lithobates
O Bidyanus
O Acipenser
O Carassius
O Oryzias
O Pimephales
O Lepomis
~ Phvsalaemus
O Salmo
• Oncorhynchus
A Asellus
O C\prinus
O Ictalurus
O Salvelinus
O Poecilia
O Xvrauchen
O Perca
+ Paratanvtarsus
O Danio
O Micropterus
A Tubifex
A Crangom'x
A Limnodrilus
A Caecidotea
A Pectinatella

0.01	0.1	1	10	100	1000

Genus Mean Acute Value (mg/L 2H-Pei fluoio-2-deceuoic acid (8:2 FTUCA))

Figure 5-7. Ranked Acute 8:2 FTUCA GMAVs Used for the Aquatic Life Acute
Benchmark Calculation.

5.9 Derivation of Acute Water Benchmark for Pentadecafluorodecanoic
Acid (7:3 IK A)

Quantitatively acceptable empirical data for 7:3 FTCA were available for two species
comprising two genera and fulfilling two MDR groups (Tables 4-2 and 5-24). Because only two
of the MDRs were met with empirical data, ICE-derived values were used to fulfill the remaining
six MDRs. A total of 70 ICE models were accepted for use. Of these, ten ICE models were not
selected for use because empirical data were available for the two genera (e.g., models for
Oncorhynchus gi/ae and Daphnia put ex). Four other models were not used because there were

74


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acceptable models for more closely-related surrogate species (i.e., Daphnia magna predicting for
Lepomis macrochirus; Daphnia magna predicting for Pimephalespromelas; Daphnia magna
predicting for Danio rerio). A total of 56 ICE models were selected for use, resulting in ICE
models predicting 55 SMAVs representing 46 genera and all eight MDRs (Tables 5-23 and 5-
24).

The combined empirical and ICE data resulted in 48 GMAVs that collectively fulfill the
eight MDR groups for deriving a freshwater criterion as defined by the Aquatic Life Criteria
Guidelines (U.S. EPA 1985). The ranked GMAVs for these combined data along with the MDR
met by each GMAV are summarized in Table 5-24. GMAVs for the four most sensitive genera
were within a factor of 1.7 of each other (Table 5-25). The freshwater FAV (the 5th percentile of
the genus sensitivity distribution) for 7:3 FTCA is 0.3727 mg/L (Table 5-25), which is lower
than all of the GMAVs except for the ICE-derived species value for the threeridge mussel,
Amblemaplicata (GMAV=0.2519 mg/L). The FAV was divided by two to obtain a preliminary
value of 0.1864 mg/L 7:3 FTCA and then adjusted by the carboxylic acid application factor
(15.8) to obtain the freshwater acute water column benchmark magnitude of 0.012 mg/L 7:3
FTCA (rounded to two significant figures). This value is expected to be protective of 95% of
freshwater genera exposed to 7:3 FTCA under short-term conditions of one-hour duration, if the
one-hour average magnitude is not exceeded more than once in three years (Figure 5-8).

There is a gap in the data between the water flea (Moina macrocopa, SMAV =1.76
mg/L) and midge (Chironomus tentans, SMAV = 4.63 mg/L) for these ICE-generated values
(see Figure 5-8). This trend was also noted for 8:2 FTUCA, the other fluorotelomer evaluated.
Although it is unclear the exact reason for this gap in values, it is noted that the lower portion of
the curve is primarily composed of the phyla Mollusca and Arthropoda, while the upper portion

75


-------
of the curve is primarily composed of the phylum Chordata. The distinct difference in sensitivity
indicated by the gap in these data is thus likely to result from a greater estimated sensitivity of
most invertebrates than vertebrates to 7:3 FTCA. Additionally, and as for 8:2 FTUCA, the ICE
estimates for the lower portion of the curve are based primarily on the surrogate Daphnia magna,
while the ICE estimates for the upper portion of the curve are based primarily on the surrogate
Oncorhynchns mykiss. However, these differences fall within the range of differences in acute
response frequently observed between species. For example, within the PFOA and PFOS acute
criteria datasets, there was single species sensitivity that spanned over two orders of magnitude,
and the variation in acute toxicity values is frequently an order of magnitude and can be as high
as three orders of magnitude (Hrovat et al. 2009, Schiir et al. 2023). It is, therefore, not
unexpected to see the difference in sensitivity observed between these species.

76


-------
Table 5-23. Acceptable Models for ICE-estimated Species Sensitivity to 7:3 FTCA.

Includes values that were extrapolated as indicated. Bold predicted EC50S used for SMAY calculations.

Predicted Species

Surrogate

ICE Model
(m

Value Range
2/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Minimum

Maximum

Amphipod (Crangonyx
pseudogracilis)

Daplinid (Daphnia magna)

0.9592

0.0008

166.8

9.5

(2.8-32.4)

Amphipod (Gammarus fasciatus)

Daplinid (Daplinia magna)

0.9592

0.0002

5,000

0.8

(0.4-1.4)

Amphipod (Gammarus
pseudolimnaeus)

Daplinid (Daplinia magna)

0.9592

0.0001

68.3

0.9

(0.4-2.2)

Apache trout (Oncorhynchus gilae)*

Rainbow trout (Oncorhynchus
mykiss)

32**

0.004

1.625

39.4
(9.0-173.4)

Atlantic salmon (Salmo salar)

Rainbow trout (Oncorhynchus
mykiss)

32

0.0002

95.9

31.0

(15.6-61.3)

Beaver-tail fairy shrimp
(Thamnocephalus platyurus)

Daplinid (Daplinia magna)

0.9592

0.0003

8,694.5

0.9

(0.6-1.3)

Bluegill (Lepomis macrocliirus)

Daplinid (Daplinia magna)

0.9592

0.0001

46,278

2.1b
(1.6-2.7)

Bluegill (Lepomis macrocliirus)

Rainbow trout (Oncorhynchus
mykiss)

32

0.0001

8,341.5

33.1

(27.6-39.6)

Bonytail (Gila elegans)

Rainbow trout (Oncorhynchus
mykiss)

32

0.003

43.07

72.7

(12.6-420.2)

Brook trout (Salvelinus fontinalis)

Rainbow trout (Oncorhynchus
mykiss)

32

0.0006

96.69

31.4

(18.8-52.5)

Brown trout (Salmo trutta)

Rainbow trout (Oncorhynchus
mykiss)

32

0.0002

95.86

33.1

(19.8-55.3)

Bryozoan (Pectinatella magnifica)

Daplinid (Daplinia magna)

0.9592**

0.014

0.232

5.3

(1.9-15.4)

Bullfrog (Lithobates catesbeianus)

Rainbow trout (Oncorhynchus
mykiss)

32

0.0006

13,400

58.8

(23.7-145.4)

Cape Fear shiner (Notropis
mekistocholas)

Rainbow trout (Oncorhynchus
mykiss)

32**

0.004

1.625

114.0

(33.0-394.0)

Channel catfish (Ictalurus punctatus)

Rainbow trout (Oncorhynchus
mykiss)

32

<0.0001

13,400

31.6

(19.9-50.1)

77


-------
Predicted Species

Surrogate

ICE Model
(m

Value Range
2/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Minimum

Maximum

Chinook salmon (Oncorhynchus
tshawytscha)*

Rainbow trout (Oncorhynchus
mykiss)

32

0.003

724.4

40.0
(24.0-66.9)

Coho salmon (Oncorhynchus
kisutch)*

Rainbow trout (Oncorhynchus
mykiss)

32

<0.0001

43.07

49.4
(38.4-63.6)

Common carp (Cyprinus carpio)

Rainbow trout (Oncorhynchus
mykiss)

32

0.0002

198.4

35.6

(16.3-77.7)

Cutthroat trout (Oncorhynchus
clarkii)*

Rainbow trout (Oncorhynchus
mykiss)

32

<0.0001

198.4

27.1
(18.7-39.3)

Cuvier's foam froglet (Physalaemus
cuvieri)

Rainbow trout (Oncorhynchus
mykiss)

32

0.7099

155.5

51.8

(30.8-87.1)

Daplinid (Ceriodaphnia dubia)

Daplinid (Daplinia magna)

0.9592

0.0003

46,278

0.6

(0.4-0.8)

Daplinid (Daplinia galeata)*

Daplinid (Daplinia magna)

0.9592**

0.0001

0.646

1.0
(0.3-3.2)

Daplinid (Daplinia longispina)*

Daplinid (Daplinia magna)

0.9592

0.0089

10.36

1.1
(0.3-4.3)

Daplinid (Daplinia pulex)*

Daplinid (Daplinia magna)

0.9592

0.0002

4,894.7

0.7
(0.5-0.9)

Daplinid (Daplinia pulicaria)*

Daplinid (Daplinia magna)

0.9592

0.014

281.6

0.7
(0.2-3.1)

Daplinid (Simocephalus serrulatus)

Daplinid (Daplinia magna)

0.9592

0.0002

7.2

0.6

(0.3-1.4)

Daplinid (Simocephalus vetulus)

Daplinid (Daplinia magna)

0.9592

0.0001

166.8

0.7

(0.1-3.8)

Fathead minnow (Pimephales
promelas)

Daplinid (Daplinia magna)

0.9592

0.0002

46,500

3.0b
(2.3-3.9)

Fathead minnow (Pimephales
promelas)

Rainbow trout (Oncorhynchus
mykiss)

32

0.0002

13,400

46.7

(36.0-60.7)

Fatmucket (Lampsilis siliquoidea)

Daplinid (Daplinia magna)

0.9592

0.014

8,694.5

0.9

(0.5-1.5)

Goldfish (Carassius auratus)

Rainbow trout (Oncorhynchus
mykiss)

32

<0.0001

13,400

62.2

(31.6-122.5)

Green floater (Lasmigona subviridis)

Daplinid (Daplinia magna)

0.9592

0.014

166.8

0.8

(0.2-3.5)

78


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Predicted Species

Surrogate

ICE Model
(m

Value Range
2/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Minimum

Maximum

Green sunfish (Lepomis cyanellus)

Rainbow trout (Oncorhynchus
mykiss)

32**

<0.0001

19.93

45.8

(19.9-105.3)

Guppy (Poecilia reticulata)

Rainbow trout (Oncorhynchus
mykiss)

32

0.0006

198.4

28.3

(7.6-105.3)

Isopod (Asellus aquaticus)

Daphnid (Daphnia magna)

0.9592

0.0003

166.8

27.3

(9.3-80.1)

Isopod (Caecidotea brevicauda)

Rainbow trout (Oncorhynchus
mykiss)

32

0.0006

82.0

8.2

(1.9-35.7)

Lake trout (Salvelinus namaycush)

Rainbow trout (Oncorhynchus
mykiss)

32

0.0002

198.4

18.0

(11.4-28.3)

Largemouth bass (Micropterus
salmoides)

Rainbow trout (Oncorhynchus
mykiss)

32

<0.0001

95.86

19.5

(9.5-40.3)

Leech (Nephelopsis obscura)

Daphnid (Daphnia magna)

0.9592

0.0008

4.369

131.2

(18.7-920.2)

Medaka (Oryzias latipes)

Rainbow trout (Oncorhynchus
mykiss)

32

0.007

54.4

52.1

(10.9-248.7)

Midge (Chironomus tentans)

Daphnid (Daphnia magna)

0.9592

0.0003

472

4.6

(0.7-32.5)

Midge (Paratanytarsus dissimilis)

Rainbow trout (Oncorhynchus
mykiss)

32

0.0006

1,330

105.8

(35.6-314.4)

Midge (Paratanytarsus
parthenogeneticus)

Daphnid (Daphnia magna)

0.9592

0.37

14,500

2.9

(0.9-9.1)

Mississippi grass shrimp
(Palaemonetes kadiakensis)

Daphnid (Daphnia magna)

0.9592

0.0003

58

0.5

(0.1-1.9)

Neosho mucket (Lampsilis
rafinesqueana)

Daphnid (Daphnia magna)

0.9592

0.042

166.8

0.8

(0.4-1.6)

Northern leopard frog (Lithobates
pipiens)

Rainbow trout (Oncorhynchus
mykiss)

32

0.002

198.4

64.7

(33.7-124.2)

Oligochaete (Limnodrilus
hoffmeisteri)

Daphnid (Daphnia magna)

0.9592

0.003

281.6

7.6

(3.1-18.5)

Oligochaete (Tubifex tubifex)

Daphnid (Daphnia magna)

0.9592

0.0001

4,894.7

10.8

(3.0-38.4)

Oriental river shrimp (Macrobrachium
nipponense)

Daphnid (Daphnia magna)

0.9592

0.011

281.6

0.4

(0.2-0.9)

79


-------
Predicted Species

Surrogate

ICE Model
(m

Value Range
2/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Minimum

Maximum

Paper pondshell (Utterbackia
imbecillis)

Daplinid (Daplinia magna)

0.9592

0.014

8,694.5

0.8

(0.5-1.3)

Pheasantshell (Ortmanniana
pectorosa)

Daphnid (Daplinia magna)

0.9592

0.042

545.9

0.7

(0.2-2.4)

Rainbow trout (Oncorhynchus
mykiss)*

Daplinid (Daplinia magna)

0.9592

0.0001

14,500

1.4
(1.2-1.8)

Razorback sucker (Xyrauchen
texanus)

Rainbow trout (Oncorhynchus
mykiss)

32

0.003

43.07

26.6

(5.3-134.0)

Shortnose sturgeon (Acipenser
brevirostruin)

Rainbow trout (Oncorhynchus
mykiss)

32

0.028

95.86

45.7

(11.2-185.6)

Silver perch (Bidyanus bidyanus)

Rainbow trout (Oncorhynchus
mykiss)

32**

0.001

7.075

73.0

(29.7-179.7)

Sockeye salmon (Oncorhynchus
nerka)*

Rainbow trout (Oncorhynchus
mykiss)

32**

0.028

7.5

25.7
(4.0-164.6)

Southern leopard frog (Lithobates
sphenocephalus)

Rainbow trout (Oncorhynchus
mykiss)

32**

0.028

9.7

312.8

(88.4-1,106.8)

Swamp lymnaea (Lymnaea stagnalis)

Daplinid (Daplinia magna)

0.9592

0.014

8,694.5

1.0

(0.4-2.9)

Tadpole physa (Physella gyrina)

Daplinid (Daplinia magna)

0.9592

0.014

8,694.5

1.0

(0.4-2.3)

Tlireeridge (Amblema plicata)

Daplinid (Daplinia magna)

0.9592

0.014

4,894.7

0.3

(0.1-0.7)

Vernal pool fairy shrimp
(Branchinecta lynchi)

Daplinid (Daplinia magna)

0.9592

0.014

8,694.5

0.8

(0.4-1.6)

Washboard (Megalonaias nervosa)

Daplinid (Daplinia magna)

0.9592

0.014

8,694.5

0.4

(0.2-0.9)

Water flea (Chydorus sphaericus)

Daplinid (Daplinia magna)

0.9592

0.009

977.6

0.4

(0.2-0.7)

Water flea (Moina macrocopa)

Daplinid (Daplinia magna)

0.9592

0.009

281.6

1.8

(0.6-5.2)

Western pearlshell (Margaritifera
falcata)

Daplinid (Daplinia magna)

0.9592

0.014

8,694.5

0.8

(0.4-1.7)

Yellow perch (Perca flavescens)

Rainbow trout (Oncorhynchus
mykiss)

32**

<0.0001

16.24

22.2

(5.1-95.7)

80


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Predicted Species

Surrogate

ICE Model
(m

Value Range
2/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Minimum

Maximum

Zebrafish (Danio rerio)

Daphnid (Daphnia magna)

0.9592

0.0001

50

3.3b
(1.1-10.0)

Zebrafish (Danio rerio)

Rainbow trout (Oncorhynchus
mykiss)

32

0.0002

229.1

17.6

(6.4-48.5)

Zebrafish-embryo (Danio rerio-
embryo)

Daphnid (Daphnia magna)

0.9592

0.0001

46,500

4.8b
(2.5-9.2)

Zebrafish-embryo (Danio rerio-
embryo)

Rainbow trout (Oncorhynchus
mykiss)

32

0.0002

8,341.5

34.4

(16.8-70.4)

*	Acceptable models that were not used because genus level empirical data were available.

*	*Measured ECso falls outside range of ICE model and predicted SMAV is based on extrapolation beyond model bounds.
a Saltwater surrogate species were not used.

b Not used because other more closely taxonomically-related models were available.

0 Not used because other more closely taxonomically-related models where the measured surrogate ECso values fell within the ICE model range were available.

81


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Table 5-24. Ranked 7:3 FTCA Genus Mean Acute Values.

Values in bold and highlighted are derived from empirical 7:3 FTCA toxicity tests with the species.

Rank

MDR
Group1

Name

Species

SMAV
(mg/L)

GMAV
(mg/L)

Percentile
Rank

1

G

Threeridge

Amblema plicata

0.2519

0.2519

0.02

2

E

Oriental river shrimp

Macrobrachium
nipponense

0.3806

0.3806

0.04

3

D

Water flea

Chydorus sphaericus

0.4278

0.4278

0.06

4

G

Washboard

Megalonaias nervosa

0.4393

0.4393

0.08

5

E

Mississippi grass
shrimp

Palaemonetes
kadictkensis

0.5324

0.5324

0.10

6

D

Daphnid

Ceriodaphnia dubia

0.6004

0.6004

0.12

7

D

Daphnid

Simocephalus serrulatus

0.5972

0.6489

0.14

D

Daphnid

Simocephalus vetulus

0.7051

8

G

Pheasantshell

Ortmanniana pectorosa

0.7379

0.7379

0.16

9

G

Western pearlshell

Margaritifera falcata

0.7857

0.7857

0.18

10

G

Neosho mucket

Lampsilis raflnesqueana

0.7949

0.8254

0.20

G

Fatmucket

Lampsilis siliquoidea

0.8571

11

G

Paper pondshell

Utterbackia imbecillis

0.8426

0.8426

0.22

12

E

Amphipod

Gammctrus fasciatus

0.7600

0.8483

0.24

E

Amphipod

Gammctrus
pseudolimncteus

0.9468

13

E

Vernal pool fairy
shrimp

Brcmchinecta lvnchi

0.8488

0.8488

0.27

14

G

Green floater

Lasmigona subviridis

0.8497

0.8497

0.29

15

E

Beaver-tail fairy
shrimp

Thamnocephcdus
platyurus

0.8773

0.8773

0.31

16

G

Tadpole physa

Physella gyrina

0.9531

0.9531

0.33

17

D

Daphnid

Daphnia magna

0.9592

0.9592

0.35

18

G

Swamp lymnaea

Lymnaea stagnalis

1.029

1.029

0.37

19

D

Water flea

Moina macrocopa

1.760

1.760

0.39

20

F

Midge

Chironomus tentans

4.630

4.630

0.41

21

H

Bryozoan

Pectinatella magnifica

5.340**

5.340

0.43

22

H

Oligochaete

Limnodrilus hoffmeisteri

7.621

7.621

0.45

23

E

Isopod

Caecidotea brevicauda

8.173

8.173

0.47

24

E

Amphipod

Crangonyx
pseudogracilis

9.512

9.512

0.49

25

H

Oligochaete

Tubifex tubifex

10.75

10.75

0.51

26

F

Midge

Paratany tarsus
dissimilis

105.75

17.45

0.53

F

Midge

Paratany tarsus
parthenogeneticus

2.880

27

B

Largemouth bass

Micropterus salmoides

19.52

19.52

0.55

28

B

Yellow perch

Perca flavescens

22.20**

22.20

0.57

29

A

Brook trout

Salve linns fontinalis

31.44

23.80

0.59

82


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Rank

MDR
Group1

Name

Species

SMAV
(mg/L)

GMAV
(mg/L)

Percentile
Rank



A

Lake trout

Salvelinus namavcush

18.01





30

B

Zebrafish

Danio rerio

24.56

24.56

0.61

31

B

Razorback sucker

Xvrctuchen texamis

26.57

26.57

0.63

32

E

Isopod

Asellus aquaticus

27.30

27.30

0.65

33

B

Guppy

Poecilia reticulata

28.31

28.31

0.67

34

B

Channel catfish

Ictalurus punctatiis

31.55

31.55

0.69

35

A

Rainbow trout

Oncorhynchus mykiss

32.00

32.00

0.71

36

A

Atlantic salmon

Salmo salar

30.96

32.02

0.73

A

Brown trout

Salmo trutta

33.12

37

B

Common carp

Cyprinus carpio

35.58

35.58

0.76

38

B

Green sunfish

Lepomis cyanellus

45.79**

38.92

0.78

B

Bluegill

Lepomis macrochirns

33.08

39

B

Shortnose sturgeon

Acipenser brevirostrum

45.67

45.67

0.80

40

B

Fathead minnow

Pimephales promelas

46.72

46.72

0.82

41

C

Cuvier's foam froglet

Phvsalaemus cuvieri

51.77

51.77

0.84

42

B

Medaka

Oryzias latipes

52.12

52.12

0.86

43

B

Goldfish

Carassius auratus

62.18

62.18

0.88

44

B

Bonytail

Gila elegans

72.68

72.68

0.90

45

B

Silver perch

Bidvanus bidvanus

73.03**

73.03

0.92

46

C

Bullfrog

Lithobates catesbeianus

58.75

106.0

0.94

C

Northern leopard frog

Lithobates pipiens

64.72

C

Southern leopard frog

Lithobates
sphenocephaly

312.8**

47

B

Cape Fear shiner

Notropis mekistocholas

114.0**

114.0

0.96

48

H

Leech

Nephelopsis obscura

131.2

131.2

0.98

**Measured EC50 falls outside range of ICE model and predicted SMAV is based on extrapolation beyond model

bounds.

1: Freshwater MDR Groups

A)	The family Salmonidae in the class Osteichthyes

B)	A second family in the class Osteichthyes, preferably a commercially or recreationally important

warmwater species (e.g., bluegill, channel catfish, etc.)

C)	A third family in the phylum Chordata (may be in the class Osteichthyes or may be an amphibian, etc.)

D)	A planktonic crustacean (e.g., cladoceran, copepod, etc.)

E)	Abenthic crustacean (e.g., ostracod, isopod, amphipod, crayfish, etc.)

F)	An insect (e.g., mayfly, dragonfly, damselfly, stonefly, caddisfly, mosquito, midge, etc.)

G)	A family in a phylum other than Arthropoda or Chordata (e.g., Rotifera, Annelida, Mollusca, etc.)

H)	A family in any order of insect or any phylum not already represented.

83


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Table 5-25. 7:3 FTCA Final Acute Value and Protective Aquatic Acute Benchmark.

Calculated Freshwater FAY based on 4 lowest values; N=48 GMAVs total





Benchmark calculated by dividing the FAY by 2 and by the carboxylic acid application factor (15.8)





GMAV









Rank

Genus

(mg/L)

ln(GMAV)

ln(GMAV)2

P=R/(N+1)

sqrt(P)

1

Amblema

0.2519

-1.38

1.90

0.020

0.143

2

Macrobrachium

0.3806

-0.97

0.93

0.041

0.202

3

Chydorus

0.4278

-0.85

0.72

0.061

0.247

4

Megalonaias

0.4393

-0.82

0.68

0.082

0.286



£ (Sum):

-4.02

4.23

0.20

0.88







P = cumulative probability











R = rank













N = number of GMAVs





S2 =

17.53



S = slope







L =

-1.923



L = X-axis intercept





A =

-0.987



A = InFAV







FAV =

0.3727











FAV/2 =

0.1864 mg/L (Preliminary Value)







Adjustment =

0.1864 / 15.8 = 0.01179 (Preliminary Value / Carboxylic Acid Application Factor)

Benchmark =

0.012 mg/L 7:3 FTCA (rounded to two significant figures)





84


-------
1.00 -T
0.90

a" 0.80 -¦
o

0.70 -

C3

O*
>

•s 0.60

# 0.50 -

S3

ct

g 0.40 4

*-w

a

I 0.30 4
(X

0.20 -¦
0.10
0.00

~

Amphibian (WeblCE)

•

Fish (Empirical)

o

Fish (WeblCE)

+

Insect (WeblCE)

A

Invertebrate (Empirical)

A

Invertebrate (WeblCE)

O

Mollusk (WeblCE)



• Preliminary Value



- Acute Benchmark

A Nephelopsis
O Notropis
~ Lithobates
O Bidvanus
O Gila
O Carassius
O Orvzias
~ Phvsalaemus
O Pim'ephales
O Acipenser
O Lepomis
O Cvprinus
O Safmo
• Oncorhynchus
O Ictalurus
O Poecilia
A Asellus
O Xwauchen
O Danio
O Salvelinus
O Perca
O Micropterus
+ Paratanytarsus
A Tubifex
A Cranjjonyx
A Caeciabtea
A Limnodrilus
A Pectinatella
4- Chironomus

A Moina
O Lvmnaea
~ Daphnia
O Physella
A Thainnocephalus
O Lasmigona
A Branchinecta
A Gammarus
O Utterbackia
O Lampsilis
O Margaritifera
O Ortmaimiana
A Simocephalus
A Ceriodaphnia
A Palaemonetes
O Meaalonaias
A Chydorus
A Macrobrachium
O Amblema

	I	I	I I I I I I	I	I	I	L

0.01	0.1	1	10	100	1000

Genus Mean Acute Value (mg/L 2H,2H,3H,3H-Pefluorodecanoic acid (7:3 FTCA))

Figure 5-8. Ranked Acute 7:3 FTCA GMAVs Used for the Aquatic Life Acute Benchmark
Calculation.

5.10 Summary of Benchmarks for Evaluated PFAS Substances and Effects
Characterization

5.10.1 Summary and Evaluation of Benchmark Values

Freshwater acute benchmarks for the eight selected PFAS are summarized in Table 5-26.

These concentrations are expected to be protective of 95% of freshwater genera exposed to these

PFAS under short-term conditions of one-hour of duration, if the one-hour average magnitude is

not exceeded more than once in three years. However, because only limited empirical toxicity

test data were available for the eight PFAS and the use of ICE models were required to fill in

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data gaps to derive the benchmarks, the recommended aquatic life acute ambient water quality
benchmarks are less certain than the standard Aquatic Life AWQC typically developed.

Quantitatively acceptable empirical toxicity data were coupled with ICE-predicted values
to try to fulfill the eight MDRs for deriving acute freshwater criteria per the Aquatic Life Criteria
Guidelines (U.S. EPA 1985). Using this approach, the eight MDRs were fulfilled for six of the
evaluated compounds (PFBA, PFHxA, PFNA, PFDA, 8:2 FTUCA, and 7:3 FTCA). Seven of the
eight MDRs were fulfilled for PFBS while six of the eight MDRs were fulfilled for PFHxS; thus,
there is somewhat greater uncertainty associated with these latter benchmarks. It is also noted
that ICE models for some chemicals were highly dependent on one or a couple of surrogate
species. For example, Daphnia magna comprised the majority of surrogate species for PFNA.
This heavy reliance on a surrogate potentially increases the uncertainty associated with these ICE
models. Conversely, there is somewhat less uncertainty associated with those PFAS benchmarks
with empirical test endpoints amongst the most sensitive test species. Notably, empirical test data
comprised the mean acute value for the most sensitive test species for PFBA (Brachionus
calyciflorus), PFHxA (Brachionus calyciflorus), and PFHxS (Danio rerio), and the second most
sensitive species for PFNA (Chydorus sphaericus). Mean acute values comprised of empirical
test data did not fall amongst the most sensitive test species for the remaining PFAS for which
acute benchmark values were derived. The resulting acute water column-based benchmark
magnitudes range from 0.012 mg/L for 7:3 FTCA to 5.3 mg/L for PFBA.

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Chemical

PFBA

PFHxA

PFNA

PFDA

PFBS

PFHxS

8:2
FTUCA

7:3
FTC A

Magnitude

5.3

4.8

0.65

0.50

5.0

0.21

0.037

0.012

Duration

One hour average

Frequency

Not to be exceeded more than once in three years on average

Table 5-27 compares the model inputs and freshwater acute benchmarks where the ICE
input PFAS value (surrogate value) was beyond the model range and the model prediction used
for deriving the EPA's acute PFAS benchmark values was based on the extrapolation approach
or using an alternative scaled approach, as discussed in Section 3 (see also Appendix F). The
alternative scaled approach adjusts the toxicity values to bring them within the ICE model range,
when necessary, by changing the measured toxicity units from mg/L to |ig/L so the values fall
within the range of ICE toxicity values, rather than allowing regressions to extrapolate beyond
the model range. This alternative approach underwent detailed evaluation by Raimondo et al.
(2024) and values were calculated using the scaled approach to compare with values calculated
using the extrapolation approach.

As shown in Table 5-27, the scaled approach allowed for the acceptance of up to two
times more models than the extrapolation approach, based primarily on the greater number of
scaled inputs and associated output values meeting the confidence interval acceptability
parameters presented in Box 1 (Section 2). Although there was a meaningful increase in the
number of models accepted with the scaled approach, there was only a slight increase in the
number of MDRs fulfilled. Using the scaled approach, MDRs were met for PFBS (an increase of
one MDR) and one additional MDR was met for PFHxS (seven versus six MDRs met). The
differences between the resulting acute benchmarks, although consistently higher using the
scaled approach, were also small, with each of the benchmarks falling within a factor of < 2.1 of

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one another, indicating close agreement between benchmark values calculated using either
approach. Raimondo et al. (2024) compared the extrapolated and scaled approaches and found
no significant differences in prediction accuracy between the two approaches. It is likely the
small but generally higher benchmark values (values were the same for PFHxS) resulting from
the scaled approach result from the increase in the number of GMAVs available using the
approach (see in Table 5-27), which will tend to mathematically increase the resulting values
because of the effect of increasing the number of genera (n) in the Aquatic Life Criteria
Guidelines calculation method.

Several investigations have indicated a trend of increasing toxicity with increasing carbon
chain length of PFAS (Cousins et al. 2020, Chambers et al. 2021). This pattern was observed for
both carboxylic acid and sulfonic acid PFAS in this document, as demonstrated by the
decreasing benchmark values with an increasing number of carbons (i.e., increasing chain
length) based on both the extrapolation and scaled approaches, as shown in Table 5-28. Further,
the final acute aquatic life criterion recently derived by the EPA (U.S. EPA 2024b) for the eight
carbon carboxylic acid PFOA is 3.1 mg/L, which falls between the benchmark values for PFHxA
with six carbons (4.8 mg/L with the extrapolated approach; 6.0 mg/L with the scaled approach)
and PFNA with nine carbons (0.65 mg/L with the extrapolated approach; 0.73 mg/L with the
scaled approach) (see Table 5-28). Similarly, the acute aquatic life criterion recently derived by
the EPA (U.S. EPA 2024c) for the eight carbon sulfonic acid PFOS is 0.071 mg/L, which falls
below the benchmark values for PFHxS with six carbons (0.21 mg/L for both the extrapolated
and scaled approaches). The trend of increasing toxicity with increasing chain length for
carboxylic acid and sulfonic acid PFAS thus holds for both the calculated acute benchmark
values and with the final acute criteria for PFOA and PFOS.

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Table 5-27. Summary of Data Inputs, MDRs Fulfilled and Benchmark Outcomes Using the Extrapolation and Scaled
Approach when Web-ICE Input PFAS Values Exceed Model Range. 		

Chemical

MDRs
Met with
Empirical
Data

Number of Accepted ICE
Predictions

MDRs Filled with ICE
Models

Total MDRs Met
(Number of GMAVs)

Acute Benchmark (mg/L)

Extrapolation
Approach

Scaled
Approach

Extrapolation
Approach

Scaled
Approach

Extrapolation
Approach

Scaled
Approach

Extrapolation
Approach

Scaled
Approach

Carboxylic Acids

PFBA

3

22

43

5

5

8(22)

8(33)

5.3

11

PFHxA

3

28

43

5

5

8(24)

8(33)

4.8

6.0

PFNA

2

29

42

6

6

8(25)

8(33)

0.65

0.73

PFDA

3

61

85

5

5

8(42)

8(60)

0.50

0.65

Sulfonic Acids

PFBS

2

55

119

5

6

7(30)

8(58)

5.0

7.6

PFHxS

2

17

18

4

5

6(12)

7(13)

0.21

0.21

Fluorotelomers

8:2
FTUCA

2

66

80

6

6

8(45)

8(55)

0.037

0.041

7:3 FTCA

2

70

81

6

6

8(48)

8(59)

0.012

0.015

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Table 5-28. Comparison of PFAS Chain Length to Perfluorocarboxylic and Perfluorosulfonic Acid Benchmarks and PFOA
and PFOS Acute Criteria Values.

Chemical

Chain Length:
Number of
Carbons

Acute Benchmark (mg/L)

PFOA and PFOS Aquatic Life
Criteria (mg/L)1

Extrapolation Approach

Scaled Approach

Carboxylic Acids

PFBA

4

5.3

11

NA

PFHxA

6

4.8

6.0

NA

PFOA

8

NA

NA

3.1

PFNA

9

0.65

0.73

NA

PFDA

10

0.50

0.65

NA

Sulfonic Acids

PFBS

4

5.0

7.6

NA

PFHxS

6

0.21

0.21

NA

PFOS

8

NA

NA

0.071

NA = Not Applicable

1 Final aquatic life criteria for PFOA and PFOS are presented in U.S. EPA (2024b) and U.S. EPA (2024c), respectively.

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5.10.2 Comparison of Benchmarks to Other Calculation Methods for Data-Limited PFAS

5.10.2.1 Comparison with Giddings et al. (2019) Approach of Combining Data Across Chemical
Class

In addition to calculating PFAS benchmarks using a combination of empirical and
predicted (ICE model) values for each individual PFAS, the EPA explored the application of an
approach used by Giddings et al. (2019) to derive protective values for data-limited synthetic
pyrethroid insecticides (detailed in Appendix G). In brief, Giddings et al. (2019) combined acute
toxicity data across several synthetic pyrethroid insecticide active ingredients (e.g., bifenthrin,
cypermethrin, permethrin, etc.) into a single pyrethroid species sensitivity distribution (SSD) to
provide a more robust dataset for the derivation of protective aquatic life values. The Giddings et
al. (2019) approach thus involved combining or "binning" data within a general chemical class,
and expected similar mode of action, to develop a larger data set for their analyses. The EPA
followed the same binning approach to estimate protective aquatic life values for PFAS, using
quantitatively acceptable empirical data (Appendix A) for combined carboxylic acid PFAS
(PFBA, PFHxA, PFNA, and PFDA) and combined sulfonic acid PFAS (PFBS and PFHxS)
datasets. The EPA additionally incorporated quantitatively acceptable empirical data used for
derivation of the acute PFOA and PFOS criteria (U.S. EPA 2024b,c) in the carboxylic acid and
sulfonic acid data groupings, respectively, to further support derivation of these benchmark
values. Data for carboxylic acid and sulfonic acid groupings were each then normalized by a
common species and pooled according to PFAS grouping to derive group-specific SSDs. Group-
specific SSDs were then generated using both the EPA's SSD Generator (as was done by
Giddings et al. 2019) and following procedures described in the Aquatic Life Criteria Guidelines
(U.S. EPA 1985). A more detailed description of the process used to derive these benchmark
values is presented in Appendix G.

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Exploratory acute benchmarks derived using the Giddings binning approach and SSD
Generator-based values are summarized in Table 5-29 and compared to those calculated using
the Aquatic Life Criteria Guidelines-based values. Exploratory benchmark values for both
carboxylic acids (5.6 times) and sulfonic acids (5.0 times) were consistently higher when
calculated using the SSD Generator-based values versus the Aquatic Life Criteria Guidelines-
based values. However, these benchmarks did not show a consistent pattern in magnitude of
differences when compared to the EPA's recommended acute benchmark values (see Table 5-
29). For carboxylic acid PFAS, for example, exploratory benchmark values calculated using the
Aquatic Life Criteria Guidelines-based approach were between 0.5 and 3.7 times different (both
higher and lower) compared to the EPA's recommended acute benchmarks. For sulfonic acid
PFAS, exploratory benchmark values calculated using the Aquatic Life Criteria Guidelines-
based values were between 0.5 and 3.0 times different (both higher and lower) compared to
recommended benchmarks. Although the differences between these outcomes cannot be
definitively determined, outcomes are likely to reflect inherent differences in the derivation
processes being used. For example, use of the EPA's Aquatic Life Criteria Guidelines procedure
to derive an SSD and calculate a protective value relies on a single distribution based on
averaging the data for the four most sensitive genera around the fifth centile of the distribution,
whereas the EPA's SSD Generator (used by Giddings et al. 2019) supports the calculation of
four different distributions (normal, logistic, triangular, and Gumbel), with the protective value
selected from the best-fit distribution. The limited size of the constituent empirical datasets
available for the binning approach used to generate SSDs based on Aquatic Life Criteria
Guidelines-based and SSD Generator-based values for the specific PFAS (24 studies total for
PFBA, PFNA, PFDA, and PFHxA datasets and seven studies total for PFBS and PFHxS

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datasets) and the limited number of datapoints available for the species used for normalization
(one to three datapoints for PFBA, PFHxA, PFNA, and PFDA datasets and seven studies total
for PFBS and PFHxS datasets) are also likely to influence the individual benchmark outcomes.

Table 5-29. Comparison of Benchmark Outcomes with Acute Benchmarks Calculated
Based on Approach by Giddings et al. (2019) (Aquatic Life Criteria Guidelines and SSD
Generator-Based Values).	

Chemical

EPA
Recommended
Acute
Benchmark
(mg/L)

Acute Benchmarks Calculated
Using Binning Approaches1

Factor Difference Between EPA
Acute Benchmark and Binning-
based Benchmark Values

Aquatic Life

Criteria
Guidelines-
Based Values
(mg/L)

SSD Generator-
Based Values
(mg/L)

Aquatic Life

Criteria
Guidelines-
Based Values

SSD Generator-
Based Values

Carboxylic Acids

PFBA

5.3

19

110

3.6

21

PFHxA

4.8

4.3

24

0.9

5.0

PFNA

0.65

0.35

2.0

0.5

3.0

PFDA

0.50

0.49

2.8

1.0

5.6

Sulfonic Acids

PFBS

5.0

15

75

3.0

15

PFHxS

0.21

0.11

0.56

0.5

2.7

1 Acute bene

imarks-binning approach is presented in Appendix G.

5.10.2.2 Comparison with Grippo et al. (2021) Approach of Deriving Water Quality-Based
Ecological Screening Values

Grippo et al. (2021) calculated aquatic life surface water ecological screening values for
each of the carboxylic acid and sulfonic acid PFAS for which the EPA developed benchmarks.
Grippo et al. (2021) employed the Great Lakes Initiative (GLI) Tier II water quality value
derivation approach (U.S. EPA 1995a,b,c), which uses secondary acute factors to account for the
increased uncertainty associated with the reduced number of tested genera, when they fall below
the eight MDRs. A comparison of the resulting screening values to the EPA's recommended

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benchmark values is presented in Table 5-30. The Tier IIGLI values calculated by Grippo et al.
(2021) using the Tier II water quality value derivation approach were generally in agreement
with the EPA benchmarks for carboxylic acid PFAS (within a factor of < 8 of one another),
whereas the Tier II GLI value calculated by Grippo et al. (2021) for sulfonic acid PFAS trended
towards a greater difference (factor of 8 for PFHxS and 130 for PFBS). This trend towards a
larger difference in Tier II GLI value versus the EPA benchmark for sulfonic acid PFAS is likely
primarily due to differences between the application factor used by the EPA for calculation of
the acute benchmark (42.3) and the application factor used by Grippo et al. (2021) for calculation
of the Tier II GLI values (13), and some differences in the data used for the calculation of the
Tier II values. If, for example, the Tier II GLI application factor of 13, used by Grippo et al.
(2021) for PFBS and PFHxS, were applied to the EPA calculated benchmarks for these
compounds (instead of the application factor used by the EPA) the difference between the two
values decreases to a factor difference of 9.2 for PFBS and 2.4 for PFHxS). Overall, the EPA's
acute benchmark values are lower than the Tier GLI values. This is primarily because the EPA
used application factors to account for the high sensitivity of some invertebrate species to PFAS,
as represented by the mayfly Neocloeon triangulifer (Soucek et al. 2023) and the cladoceran
Moina micrura (Razak et al. 2023) to PFOA and PFOS. These sensitive species were not
represented in the derived Tier II GLI values.

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Table 5-30. Comparison of Benchmark Outcomes with Water Quality-Based Ecological
Screening Values Calculated by Grippo et al. (2021) Using the Great Lakes Initiative (GLI)

Chemical

EPA Recommended
Acute Benchmark
(mg/L)

Grippo et al. (2021)
Calculated Tier II
GLI Values
(mg/L)

Factor Difference
Between EPA Acute
Benchmark and
Calculated Tier II GLI
Values

Carboxylic Acids

PFBA

5.3

13.75

2.6

PFHxA

4.8

17.5

3.6

PFNA

0.65

2.14

3.3

PFDA

0.50

4

8

Sulfonic Acids

PFBS

5.0

149.1

30

PFHxS

0.21

1.7

8

5.10,3 Summary and Conclusions

The EPA developed recommended aquatic life acute ambient water quality benchmarks

for eight PFAS with limited empirical test data in accordance with the provisions of section
304(a) of the CWA. Benchmarks were developed for the following eight chemicals:
Perfluorobutanoic acid (PFBA), Perfluorohexanoic acid (PFHxA), Perfluorononanoic acid
(PFNA), Perfluorodecanoic acid (PFDA), Perfluorobutanesulfonic acid (PFBS),
Perfluorohexanesulfonic acid (PFHxS), 2H-Perfluoro-2-decenoic acid (8:2 FTUCA), and
2H,2H,3H,3H-Pefluorodecanoic acid (7:3 FTCA).

Because of limitations in the available direct empirical test data, the EPA developed acute
protective benchmarks for these PFAS using available freshwater species empirical test data in
conjunction with the application of the EPA, Office of Research and Development's (ORD's)
peer-reviewed Web-based Interspecies Correlation Estimate tool (Web-ICE; Version 4.0;
https://www.epa.gov/webice/) (Raimondo et al. 2024). Two approaches were used for

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benchmark development because PFAS acute values can extend beyond the range of values used
to calibrate the ICE model. One approach allowed extrapolation of the data beyond the calibrated
range of the model, while the other (presented in Appendix F) "scaled" the toxicity values to
bring them within the ICE model range, when necessary.

Table 5-27 summarizes and compares the model inputs and aquatic life ambient water
quality benchmark values between the extrapolation approach and alternative scaled approach.
Benchmark values calculated using the scaled approach were consistently higher than those
calculated using the extrapolation approach, though the difference was small (within a factor of
<2.1 of one another), indicating close agreement between the two approaches. It is likely this
small but consistent difference results from an increase in the number of GMAVs available when
using the scaled approach, which tends to mathematically increase the resulting values.

Several investigations have indicated a trend of increasing toxicity with increasing carbon
chain length of PFAS compounds (Cousins et al. 2020, Chambers et al. 2021). This pattern was
also observed for carboxylic acid and sulfonic acid PFAS, as demonstrated by the decreasing
benchmark values with an increasing number of carbons (i.e., increasing chain length) for both
the extrapolation and scaled approaches (Table 5-28).

Table 5-31 compares the EPA benchmark values with those calculated using alternative
approaches. The EPA explored the application of an alternative data "binning" approach,
developed by Giddings et al. (2019) (see Table 5-31), which combines data for multiple related
compounds to provide a more robust dataset. This approach supported derivation of protective
aquatic life values for grouped perfluoroalkyl carboxylic acids and grouped perfluoroalkyl
sulfonic acids. SSDs were derived using both Aquatic Life Criteria Guidelines -based
procedures, which derive a single distribution and determine the benchmark value based on

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averaging the data for the four most sensitive genera around the fifth centile, and by using the
EPA's SSD Generator, which was used by Giddings et al. (2019) and supports the calculation of
four different distributions and selects a protective value from the best-fit distribution using all of
the genera. Exploratory benchmark values for both carboxylic acids and sulfonic acids were
consistently higher (5.6 to 5.8 times and 5.0 times, respectively) when derived using the SSD
Generator versus the Aquatic Life Criteria Guidelines procedures but did not show a consistent
pattern in magnitude when compared to the EPA's recommended benchmark values (Tables 5-
31 and 5-29). For carboxylic acid PFAS, for example, exploratory benchmark values calculated
using Aquatic Life Criteria Guidelines-based values were between 1.02 and 3.6 times different
(both higher and lower) compared to EPA's recommended acute benchmarks. For sulfonic acid
PFAS, exploratory benchmark values calculated using Aquatic Life Criteria Guidelines-based
values were between 1.9 and 3.0 times different (both higher and lower) compared to
recommended benchmarks. These mostly small differences likely reflect inherent differences in
the derivation processes being used and the constituent datasets.

Grippo et al. (2021) employed the Great Lakes Initiative (GLI) Tier II water quality value
derivation approach (U.S. EPA 1995a,b,c) to derive draft ecological screening values for the
same carboxylic acid and sulfonic acid PFAS evaluated in this document. This GLI approach
uses secondary acute factors to account for the increased uncertainty associated with the reduced
number of tested genera, when the number of genera for which data is available falls below the
Aquatic Life Criteria Guidelines eight MDRs. The Grippo et al. (2021) calculated draft Tier II
GLI values were in generally close agreement with the EPA's acute benchmarks for both
carboxylic acid PFAS (within a factor of < 8 of one another) and sulfonic acid PFAS (30 for
PFBS and 8 for PFHxS) (Table 5-31). These differences likely reflect the use of different

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application factors and some differences in the data used for the calculation of the draft Grippo et
al. (2021) Tier II values, which did not include the sensitive mayfly (N. Iriangiilifer) data as the
Grippo et al. (2021) publication preceded the publication of the mayfly data.

These acute aquatic life benchmarks, developed under Section 304(a)(2) of the CWA,
are informational values that the EPA generated because there are limited high quality toxicity
data available and data gaps exist for several aquatic organism families, such that 304(a)(1)
criteria could not be developed. The EPA developed these aquatic life benchmarks to provide
information that states and Tribes may consider in their water quality protection programs,
including establishing their water quality criteria. However, the EPA's aquatic life benchmark
values are not regulatory, nor do they automatically become part of a state's or Tribe's water
quality standards.

Table 5-31. Summary of the EPA Acute Benchmark Values and Acute Values Calculated
Using Alternative Methods 		

Chemical

EPA Acute
Benchmarks -
Extrapolation
Approach
(mg/L)

Acute Benchmarks
Binning A

Calculated Using
pproaches

Grippo et al (2021)
Calculated Draft Tier
II GLI Values
(mg/L)

Aquatic Life Criteria
Guidelines-Based
Values (mg/L)

SSD Generator-
Based Values
(mg/L)

PFBA

5.3

19

110

13.75

PFHxA

4.8

4.3

24

17.5

PFNA

0.65

0.35

2.0

2.14

PFDA

0.50

0.49

2.8

4

PFBS

5.0

15

75

149.1

PFHxS

0.21

0.11

0.56

1.7

8:2 FTUCA

0.037

-

-

-

7:3 FTCA

0.012

-

-

-

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for machine learning in ecotoxicology. Sci. Data. 10:718. https://doi.org/10.1038/s41597-Q23-
02612-2

Soucek, D.J., R.A. Dorman, E.L. Pulster, B.G. Perrotta, D.M. Walters and J.A. Steevens. 2023.
Perfluorooctanesulfonate adversely affects a mayfly {Neocloeon tricmgulifer) at environmentally
realistic concentrations. Environ. Sci. Technol. Lett. DOI: DOI: 10.1021/acs.estlett.3c00056
<3March2023>.

U.S. EPA (United States Environmental Protection Agency). 1985. Guidelines for deriving
numerical national water quality criteria for the protection of aquatic organisms and their uses.
EPA Report 822/R-85-100 (NTIS Report PB85-227049).

U.S. EPA (United States Environmental Protection Agency). 1995a. Final water quality guidance
for the Great Lakes system. 60 CFR Part 60, No. 56.

U.S. EPA (United States Environmental Protection Agency). 1995b. Water quality guidance for
the Great Lakes system. 40 CFR Part 132.

U.S. EPA (United States Environmental Protection Agency. 1995c. Great Lakes water quality
initiative technical support document for wildlife criteria. EPA-820-B-95-009, Office of Water,
Washington, D.C.

U.S. EPA (United States Environmental Protection Agency. 2016. OCSPP 850.1000 Background
and Special Considerations-Tests with Aquatic and Sediment-Dwelling Fauna and Aquatic
Microcosms (EPA-HQ-OPPT-2009-0154-0042). Office of Chemical Safety and Pollution
Prevention. Washington, D.C.

U.S. EPA (United States Environmental Protection Agency). 2018. Final aquatic life ambient
water quality criteria for aluminum - 2018 (EPA-822-R-18-001). Office of Water. Washington,
D.C.

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U.S. EPA (United States Environmental Protection Agency). 2022a. Draft aquatic life ambient
water quality criteria for perfluorooctanoic acid (PFOA). EPA-842-D-22-001. U.S. EPA, Office
of Water, Washington, D.C.

U.S. EPA (United States Environmental Protection Agency). 2022b. Draft Aquatic Life Ambient
Water Quality Criteria for Perfluorooctane Sulfonate (PFOS). EPA-842-D-22-002. U.S. EPA,
Office of Water, Washington, D.C.

U.S. EPA (United States Environmental Protection Agency). 2024a. Method 1633 analysis of
per- and polyfluoroalkyl substances (PFAS) in aqueous, solid, biosolids, and tissue samples by
LC-MS/MS. EPA 821-R-24-001. U.S. EPA, Office of Water, Washington, D.C.

U.S. EPA (United States Environmental Protection Agency). 2024b. Freshwater aquatic life
ambient water quality criteria and acute saltwater aquatic life benchmark for perfluorooctanoic
acid (PFOA). EPA-842-R-24-002. U.S. EPA, Office of Water, Washington, D.C.

U.S. EPA (United States Environmental Protection Agency). 2024c. Freshwater aquatic life
ambient water quality criteria and acute saltwater aquatic life benchmark for perfluorooctane
sulfonate (PFOS). EPA-842-R-24-003. U.S. EPA, Office of Water, Washington, D.C.

Wang, W.J., C. DeWitt, C.P. Higgins and I.T. Cousins. 2017. A never-ending story of per- and
polyfluoroalkyl substances (PFASs)? Environ. Sci. Technol. 5(51): 2508-2518.

Wang, X., B. Fan, M. Fan, S. Belanger, J. Li, J. Chen, X. Gao and Z. Liu. 2020. Development
and use of interspecies correlation estimation models in China for potential application in water
quality criteria. Chemosphere. 240: 124848.

Willming, M.M., C.R. Lilavois, M.G. Barron and S. Raimondo. 2016. Acute toxicity prediction
to threatened and endangered species using interspecies correlation estimation (ICE) models.
Environ. Sci. Technol. 50: 10700-10707.

Wu, J., Z. Liu, Z. Yan and X. Yi. 2015. Derivation of water quality criteria of phenanthrene
using interspecies correlation estimation models for aquatic life in China. Environ. Sci. Pollut.
Res. 22: 9457-9463.

Wu, J., Z. Yan, X. Yi, Y. Lin, J. Ni, X. Goa., Z. Liu and X. Shi. 2016. Comparison of species
sensitivity distributions constructed with predicted acute toxicity data from interspecies
correlation estimation models and measured acute data for Benzo[a]pyrene. Chemosphere. 144:
2183-2188.

Zhang, S., L. Wang, Z. Wang, D. Fan, L. Shi and J. Lui. 2017. Derivation of freshwater water
quality criteria for dibutyltin dilaurate from measured data and data predicted using interspecies
correlation estimation models. Chemosphere. 171: 142-148.

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Appendices

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Appendix A Quantitatively Acceptable Freshwater Acute Toxicity Studies

Species (lifestage)

Method"

Test
Duration

Chemical /
Purity

i)H

Temp.

(°C)

Effect

Author Reported
Effect Cone.
(mg/L)

Species Mean
Acute Value
(mg/L)

Reference

Perfluorobutanoic acid (PFBA)

Rotifer (neonate, <2 h),

Brachionus calvciflorus

S,U

24 hr

PFBA

98%

7.0

20

LCso

110

110

Wang et al. 2017



Cladoceran (neonate, <24
hours), Chvdorus sphaericus

S, M

48 hr

PFBA

98%

-

20

ECso

(immobilization)

>4,280.8

>4,280.8

Ding et al. 2012



Zebrafish (embryo, 60 mpf),

Danio rerio

S,U

96 hr

PFBA

>97%

7.2-
7.5

26

LCso

>3,000b

-

Hagenaars et al. 2011

Zebrafish (gastrula, 4.5 hpf),

Danio rerio

R, U

96 hr

PFBA
NR

7.0 -
7.5

28

LCso

13,779

13,779

Godfrey et al. 2017



Cladoceran (neonate, <24
hr), Daphnia magna

S,U

48 hr

PFBA

>97%

NR

21

ECso

(immobilization)

>l,006b

-

Boudreau 2002

Cladoceran (neonate, <24
hr), Daphnia magna

S, M

48 hr

PFBA

98%

8.0

20

ECso

(immobilization)

>4,280.8

-

Ding et al. 2012

Cladoceran (neonate, <24
hr), Daphnia magna

S, M

48 hr

PFBA

>98%

7.0-
7.82

20

ECso

(immobilization)

5,251

4,741

Bannentlo et al. 2015



Cladoceran (neonate, <24
hr), Daphnia pulicaria

S,U

48 hr

PFBA

>97%

-

21

ECso

(immobilization)

>1,006

>1,006

Boudreau 2002

Perfluorobutanesulfonic acid (PFBS)

Cladoceran (neonate, <24
hr), Daphnia magna

S, M

48 hr

PFBS
97.3-97.9%

8.1-
8.5

19.8-
20.1

ECso

(immobilization)

2,183

2,183

Wildlife International Ltd.
2001



Fathead minnow (juvenile),

Pimephales promelas

S, M

96 hr

PFBS

97.3 - 97.9%

8.4-
8.5

20.7-
22.8

LCso

1,938

1,938

Wildlife International Ltd.
2001



Bluegill (juvenile), Lepomis
macrochirus

S, M

96 hr

PFBS

97.3 - 97.9%

8.1-
8.4

22.1-
23.9

LCso

6,452

6,452

Drottar and Krueger 2001



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Species (lifestage)

Method3

Test
Duration

Chemical /
Purity

pH

Temp.

(°C)

Effect

Author Reported
Effect Cone.
(mg/L)

Species Mean
Acute Value
(mg/L)

Reference

Zebrafish (embryo, 60 mpf),

Danio rerio

S,U

96 lir

PFBS

>97%

7.2-
7.5

26

LCso

>3,000

>3,000

Hagenaars et al. 2011

Perfluorononanoic acid (PFNA)

Cladoceran (<24 hours),
Chvdorus sphaericus

S, M

48 lir

PFNA
97%

-

20

ECso

(immobilization)

27.84

27.84

Ding et al. 2012



Cladoceran (neonate, 24
hours), Daphnia magna

S,U

48 lir

PFNA
98%

7.7-
8.4

20

ECso

(immobilization)

43.42

-

Lu et al. 2016

Cladoceran (neonate, 24
hours), Daphnia magna

S,U

48 lir

PFNA
>97%

-

21

ECso

(immobilization)

91.89

-

Boudreau 2002

Cladoceran (neonate, 24
hours), Daphnia magna

S, M

48 lir

PFNA
97%

-

20

ECso

(immobilization)

151.29

84.51

Ding et al. 2012



Clawed frog (blastula),

Xenopus sp.

R, U

96 lir

PFNA
NR

-

23

LCso

335.8

335.8

Kim et al. 2013

Perfluorodecanoic acid (PFDA)

Cladoceran (neonate, <24
hours), Chvdorus sphaericus

S,U

48 lir

PFDA

98%

-

20

ECso

(immobilization)

41.13

41.13

Ding et al. 2012



Cladoceran (neonate, <24
hours), Daphnia magna

S,U

48 lir

PFDA

99.6%

-

20

ECso

>100b

-

Hoke et al. 2012

Cladoceran (neonate, <24
hours), Daphnia magna

S,U

48 lir

PFDA

>97%

-

21

ECso

(immobilization)

129.54

-

Boudreau 2002

Cladoceran (neonate, <24
hours), Daphnia magna

s,u

48 lir

PFDA

-

20

ECso

(immobilization)

163.48

145.5

Ding et al. 2012



Cladoceran (neonate, <24
hours), Daphnia pulicaria

s,u

48 lir

PFDA,

>97%

-

21

ECso

(immobilization)

149.59

149.59

Boudreau 2002



Clawed frog (blastula),

Xenopus sp.

R, U

96 lir

PFDA
NR

-

23

LCso

76.5

76.5

Kim et al. 2013



A-2


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Species (lifestage)

Method3

Test
Duration

Chemical /
Purity

pH

Temp.

(°C)

Effect

Author Reported
Effect Cone.
(mg/L)

Species Mean
Acute Value
(mg/L)

Reference

Rainbow trout (not reported,
28-61 dph), Oncorhynchus
mvkiss

S,U

96 lir

PFDA

99.6%



12

LCso

32

32

Hoke et al. 2012

Perfluorohexanoic acid (PFHxA)

Rotifer (neonate, <2 h),

Brachionus calvciflorus

s,u

24 lir

PFHxA
98%

7.0

20

LCso

140

140

Wang et al. 2017



Cladoceran (neonate, <24
hours), Daphnia magna

S, M

48 lir

PFHxA
>97%

7.0-
7.82

20

ECso

(immobilization)

1,048

1,048

Bannentlo et al. 2015



American bullfrog (larva, 26
go), Lithobates catesbeianus
(formerly, Rana catesbeiana)

S,U

96 lir

PFHxA
>98%

-

21

LCso

1,105

1,105

Tornabene et al. 2021



Green frog (larva, 26 gosner
stage), Lithobates clamitans
(formerly, Rana clamitans)

S,U

96 lir

PFHxA
NR

-

21

LCso

758

758

Tornabene et al. 2021

Perfluorohexanesulfonic acid (PFHxS)

Zebrafish (embryo, 3 hpf),

Danio rerio

RU

96 lir

PFHxS
NR

7.2

28

NOEC

(growth, length)

22.5

22.5

Annunziato et al. 2020



American bullfrog (larva, 26
gosner stage), Lithobates
catesbeianus

(formerly, Rana catesbeiana)

S,U

96 lir

PFHxS
NR

-

21

LCso

1,105

1,105

Tornabene et al. 2021



Green frog (larva, 26 gosner
stage), Lithobates clamitans
(formerly, Rana clamitans)

S,U

96 lir

PFHxS
NR

-

21

LCso

758

758

Tornabene et al. 2021

H-Perfluoro-2-decenoic acid (8:2 FTUCA)

Cladoceran (neonate, 24 lir),

Daphnia magna

s,u

48 lir

8:2 FTUCA
99.8%

-

20

ECso

(immobilization)

3.2

3.2

Hoke et al. 2012

A-3


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Species (lifestage)

Method3

Test
Duration

Chemical /
Purity

pH

Temp.

(°C)

Effect

Author Reported
Effect Cone.
(mg/L)

Species Mean
Acute Value
(mg/L)

Reference

Rainbow trout (not reported,
28-61 dph), Oncorhynchus
mvkiss

S,U

96 hr

8:2 FTUCA
99.8%



12

LCso

81

81

Hoke et al. 2012

2H,2H,3H,3H-Pefluorodecanoic acid (7:3 FTCA)

Cladoceran (neonate, <24
hr), Daphnia magna

S, M

48 hr

7:3 FTCA

>99.%

-

20

ECso

(immobilization)

0.4

-

Hoke et al. 2012

Cladoceran (neonate, <24
hr), Daphnia magna

S,U

48 hr

7:3 FTCA
>99%

-



ECso

(immobilization)

2.3

0.9592

Hoke et al. 2012



Rainbow trout (not reported,
28-61 dph), Oncorhynchus
mvkiss

S,U

96 hr

7:3 FTCA
>99%

-

12

LCso

32

32

Hoke et al. 2012

a S=static, R=renewal, F=flow-through, U=unmeasured, M=measured.
b Not used in species mean calculation because the value is greater than low value.

A-4


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A.l References

Annunziato, K.M., J. Doherty, J. Lee, J.M. Clark, W. Liang, C.W. Clark, M. Nguyen, M.A. Roy
and A.R. Timme-Laragy. 2020. Chemical characterization of a legacy aqueous film-forming
foam sample and developmental toxicity in zebrafish (Danio rerio). Environ. Health
Perspect. 128(9): 97006-97006.

Barmentlo, S.H., J.M. Stel, M. Van Doom, C. Eschauzier, P. De Voogt and M.H.S. Kraak. 2015.
Acute and chronic toxicity of short chained perfluoroalkyl substances to Daphnia magna.
Environ. Pollut. 198:47-53.

Boudreau, T.M. 2002. Toxicity of perfluorinated organic acids to selected freshwater organisms
under laboratory and field conditions. M.S. Thesis, University of Guelph, Ontario, Canada: 145
P-

Ding, G.H., T. Fromel, E.J. Van den Brandhof, R. Baerselman and W.J.G.M. Peijnenburg. 2012.
Acute toxicity of poly- and perfluorinated compounds to two cladocerans, Daphnia magna and
Chydorus sphaericus. Environ. Toxicol. Chem. 31(3): 605-610.

Drottar, K.R. and H.O. Krueger. 2001. Perfluoro butane sulfonate, potassium salt (PFBS): A 96-
hour static acute toxicity test with bluegill (Lepomis macrochirus). Project 454A-114. Wildlife
International Ltd.:43 p.

Godfrey, A., A. Abdel-Moneim and M.S. Sepulveda. 2017. Acute mixture toxicity of
halogenated chemicals and their next generation counterparts on zebrafish embryos.
Chemosphere. 181:710-712.

Hagenaars, A., L. Vergauwen, W. De Coen and D. Knapen. 2011. Structure-activity relationship
assessment of four perfluorinated chemicals using a prolonged zebrafish early life stage test.
Chemosphere. 82:764-772.

Hoke, R.A., L.D. Bouchelle, B.D. Ferrell and R.C. Buck. 2012. Comparative acute freshwater
hazard assessment and preliminary PNEC development for eight fluorinated acids.

Chemosphere. 87(7): 725-733.

Kim, M., J. Son, M.S. Park, Y. Ji, S. Chae, C. Jun, J.S. Bae, T.K. Kwon, Y.S. Choo, H. Yoon, D.
Yoon, J. Ryoo and S.H. Kim. 2013. In vivo evaluation and comparison of developmental toxicity
and teratogenicity of perfluoroalkyl compounds using Xenopus embryos. Chemosphere. 93(6):
1153-1160.

Lu, G.H., B.H. Ma, S. Li, and L.S. Sun. 2016. Toxicological Effects of Perfluorooctanoic Acid
(PFOA) on Daphnia magna. Mat. Sci. Environ. Eng.: 559-564

A-5


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Tornabene, B.J., M.F. Chislock, M.E. Gannon, M.S. Sepulveda and J.T. Hoverman. 2021.
Relative acute toxicity of three per- and polyfluoroalkyl substances on nine species of larval
amphibians. Integr. Environ. Assess. Manag.l7(4): 684-689.

Wang, Y., J. Niu, L. Zhang and J. Shi. 2014. Toxicity assessment of perfluorinated carboxylic
acids (PFCAs) towards the rotifer Brachionus calyciflorus. Sci. Total Environ. 491/492:266-270.

Wang, W.J., C. DeWitt, C.P. Higgins and I.T. Cousins. 2017. A never-ending story of per- and
polyfluoroalkyl substances (PFASs)? Environ. Sci. Technol. 5(51): 2508-2518.

Wildlife International Ltd.. 2001. Perfluorobutane Sulfonate, Potassium Salt: (PFBS): A 48-hour
static acute toxicity test with the cladoceran (Daphnia magna). Wildlife International Ltd.

Project No. 454A-118A: 42 p.

A-6


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Appendix B Summary of Lowest Quantitatively Acceptable Freshwater
Acute Toxicity Studies

B.l Summary of Lowest Quantitatively Acceptable Empirical Toxicity Studies Used to

Derive Aquatic Life Benchmarks

Quantitatively acceptable empirical acute toxicity data were available for 27 freshwater
species across the 8 PFAS: PFBA (5 species), PFBS (4 species), PFNA (3 species), PFDA (4
species), PFHxA (4 species), PFHxS (3 species), 8:2 FTUCA (2 species), and 7:3 FTCA (2
species) (Table 4-2; Appendix A). The following presents summaries of the freshwater toxicity
studies with empirical effect values that were used quantitatively to derive acute benchmarks,
which consist of toxicity values falling amongst the four most sensitive values. These values are
used quantitatively to derive the benchmarks consistent with procedures presented in the Aquatic
Life Criteria Guidelines (U.S. EPA 1985) designed to protect 95% of aquatic taxa. These "most
sensitive" empirical studies consist of one study each for PFBA, PFNA, PFHxA, and PFHxS.
Study summaries for the most sensitive taxa are presented below for each of these substances.
Acute values are presented as reported by the study authors, unless indicated otherwise.

B.2 Summary of Lowest Quantitatively Acceptable Empirical Toxicity Studies for

Perfluorobutanoic acid (PFBA)

Wang et al. (2014) exposed the rotifer, Brachionus calyciflorus, to perfluorobutanoic
acid (PFBA, CAS # 375-22-4, 98% purity) in a 24-hour acute toxicity test. The test was
performed under static, non-renewal conditions beginning with <2-hour old neonates. Exposures
consisted of 10 test organisms per 15 mL well in 6-well cell culture plates containing 10 mL of
test medium. The control and each of nine PFBA test concentrations were replicated three times
for a total of 30 neonates per treatment. Nominal test concentrations were 0.0 mg/L for the
control and treatments ranged from 40-140 mg/L PFBA. Neonates were exposed at pH 7.0 and
20 ± 1°C under darkness. After 24 h of exposure, the number of dead and live rotifers was

B-l


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enumerated under a microscope at 10x magnification. Rotifers were deemed inactive if they did
not move in a period of 30s. The author-reported acute LC50 was 110 mg/L. This value was
acceptable for quantitative use in deriving the recommended acute freshwater PFBA benchmark.

B.3 Summary of Lowest Quantitatively Acceptable Empirical Toxicity Studies for
Perfluorohexanoic acid (PFHxA)

Wang et al. (2014) exposed the rotifer, Brachionus calyciflorus, to perfluorohexanoic
acid (PFHxA, CAS # 307-24-4, 98% purity) in a 24-hour acute toxicity test. The test was
performed under static, non-renewal conditions beginning with <2-hour old neonates. Exposures
consisted of 10 test organisms per 15 mL well in 6-well cell culture plates containing 10 mL of
test medium. The control and each of nine PFHxA test concentrations were replicated three times
for a total of 30 neonates per treatment. Nominal test concentrations were 0.0 mg/L for the
control and treatments ranged from 40-140 mg/L PFHxA. Neonates were exposed at pH 7.0 and
20 ± 1°C under darkness. After 24 h of exposure, the number of dead and live rotifers was
enumerated under a microscope at 10x magnification. Rotifers were deemed inactive if they did
not move in a period of 30s. The author-reported acute LC50 was 140 mg/L. This value was
acceptable for quantitative use in deriving the recommended acute freshwater PFHxA
benchmark.

B.4 Summary of Lowest Quantitatively Acceptable Empirical Toxicity Studies for
Perfluorononanoic acid (PFNA)

Ding et al. (2012) exposed the cladoceran, Chydorus sphaericus, to perfluorononanoic
acid (PFNA, CAS # 375-95-1, 97% purity) in a 48-hour acute toxicity test. The test was
performed under static, non-renewal conditions beginning with <24-hour old neonates.

Exposures consisted of five test organisms per 2 ml vial to which 250 |il test solution was added.
Because the water solubility of PFNA was low, dimethylsulfoxide (DMSO) was used for
preparing the stock solution. The concentration of DMSO in test solutions did not exceed 0.2%.

B-2


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A solvent control was tested simultaneously with a negative (dilution water) control. The two
controls and each of six PFNA test concentrations were replicated four times for a total of 20
neonates per treatment. Nominal test concentrations were 0.0 (controls), 0.05, 0.07, 0.1, 0.15, 0.2
and 0.25 mMPFNA, or 0, 23.20, 32.49, 69.61, 92.82 and 116.0 mg/L after conversion using a
molecular weight for PFNA of 464.08 mg/mmol. Samples containing the highest and the lowest
test concentrations were analyzed to confirm the nominal concentrations. Although partial
chemical analysis was conducted, only nominal concentrations were reported. To assess the test
concentration variability, all samples at a selected nominal concentration were analyzed, with a
relative standard deviation of 6.5%. The measured concentrations were between 82 and 91% of
the nominal concentration. Neonates were exposed at 20 ± 1°C under a 16:8-hour light:dark
cycle. The animals were not fed during the test period and were checked under a reverse
dissecting microscope at 24 and 48 h. Immobilization was determined based on inactivation of
the animals after slightly tapping with a finger to the vial and monitoring them for 30s. The
author-reported acute immobilization LCso was 0.060 mM (or 27.84 mg/L). This value was
acceptable for quantitative use in deriving the recommended acute freshwater PFNA benchmark.

B.5 Summary of Lowest Quantitatively Acceptable Empirical Toxicity Studies for

Perfluorohexanesulfonic acid (PFHxS)

Annunziato et al. (2020) exposed the wildtype AB strain of zebrafish, Danio rerio, to
perfluorohexanesulfonic acid (PFHxS, CAS # 355-46-4, purity not reported) in a 93-hour acute
toxicity test following OECD Test No. 236 (OECD 2013). The test was performed under static-
renewal conditions beginning with 3-hour post fertilization (hpf) embryos. Fifteen embryos were
assigned at random to each treatment or control group, with each embryo placed into an
individual glass vial containing 1 mL of solution. Test solutions were prepared and refreshed
daily. Because of the low water solubility of PFHxS, dimethylsulfoxide (DMSO) was used for

B-3


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preparing the stock solution. The concentration of DMSO in test solutions did not exceed 0.01%.
A solvent control and each of seven PFHxS test concentrations were replicated three times for a
total of 45 organisms per treatment. Nominal test concentrations were 0.0 (control), 0.07, 1.41,
2.81, 5.63, 11.25, 22.50, and 45.00 mg/L PFHxS. Embryos were exposed at pH 7.2, 28 ± 1°C
under a 14:10-hour light:dark cycle and monitored daily through 96 hpf. There was >90% health
and survival in the control group. Test concentrations of PFHxS were too low to derive an LCso
value. Larval growth (length) was assessed at test termination. The author-reported a growth
(weight) NOEC of 22.5 mg/L. This value was acceptable for quantitative use in deriving the
recommended acute freshwater PFHxS benchmark.

B.6 References

Annunziato, K.M., J. Doherty, J. Lee, J.M. Clark, W. Liang, C.W. Clark, M. Nguyen, M.A. Roy
and A.R. Timme-Laragy. 2020. Chemical characterization of a legacy aqueous film-forming
foam sample and developmental toxicity in zebrafish (Danio rerio). Environ. Health
Perspect. 128(9): 97006-97006.

Ding, G.H., T. Fromel, E.J. Van den Brandhof, R. Baerselman and W.J.G.M. Peijnenburg. 2012.
Acute toxicity of poly- and perfluorinated compounds to two cladocerans, Daphnia magna and
Chydorus sphaericus. Environ. Toxicol. Chem.31(3): 605-610.

U.S. EPA (United States Environmental Protection Agency). 1985. Guidelines for deriving
numerical national water quality criteria for the protection of aquatic organisms and their uses.
EPA Report 822/R-85-100 (NTIS Report PB85-227049).

Wang, Y., J. Niu, L. Zhang and J. Shi. 2014. Toxicity assessment of perfluorinated carboxylic
acids (PFCAs) towards the rotifer Brachionus calyciflorus. Sci. Total Environ. 491/492: 266-
270.

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Appendix C Qualitative Freshwater Acute Toxicity Studies

Species (lifestage)

Method3

Test
Duration

Chemical /
Purity

PH

Temp.
(°C)

Effect

Author
Reported
Effect
Concentration
(mg/L)

Deficiencies

Reference

Perfluorobutanoic acid (PFBA)

Green alga, Raphidocelis
subcapitata

(formerly, Pseudokirchneriella
subcapitata and Selenastrum
capriconnituni)

S,U

4.5 hr

Perfluorobutanoic
acid/

98%

NR

NR

ECso
(population
photosynthesis)

262.2b

Test duration too
short

Ding et al. 2012



Duckweed,
Lenma gibba

s,u

7 d

Perfluorobutyric
acid/

>97%

NR

25

IC50

(population/
weight)

>l,006b

Test duration too
long

Boudreau 2002



Japanese medaka (embryo, <6

hpf),

Orvzias latipes

R,U

48 hr post
hatch

Perfluorobutyric
acid/

NR

7.0-

7.5

25

LOEC
(velocity)

137

Only one
concentration
tested; atypical
endpoint

Godfrey 2017

Japanese medaka,
Orvzias latipes

S,U

96 hr

Perfluorobutyric
acid/

NR

7.0-

7.5

25

NOEC
(mortality)

>13,795

Only one

concentration tested

Godfrey et al. 2019

Japanese medaka (embryo, <6

hpf),

Orvzias latipes

R,U

10 d

Perfluorobutyric
acid/

NR

7.0-

7.5

25

NOEC
(growth -
length)

137

Only one
concentration
tested; duration too
short for a chronic
test

Godfrey et al. 2019



Zebrafish (embryo, 6-8 hpf),
Danio retio

S,U

144 hr
post fert.

Perfluorobutyric
acid/

NR

7.2-

7.6

26

ECso
(heart rate,
hatching time)

2,200

Duration too long
for an acute test and
too short for a
chronic test

Ulhaqetal. 2013

Zebrafish (embryo, 5-6 hpf),
Danio retio

R, U

90-91 hr

Perfluorobutyric
acid/

>99%

7

28

LCso

9,703

Test duration too
short

Wasel et al. 2020

C-l


-------
Species (lifestage)

Method3

Test
Duration

Chemical /
Purity

pH

Temp.
(°C)

Effect

Author
Reported
Effect
Concentration
(mg/L)

Deficiencies

Reference

Zebrafish (embryo, 6-8 hpf),
Danio retio

S,U

112-114
hr

Perfluoro-n-
butanoic acid /

>98%

NR

28

NOEC
(mortality)

0.2504b

Only one
concentration
tested; duration too
long for an acute
test and too short for
a chronic test

Rericha et al. 2021

Perfluorobutanesulfonic acid (PFBS)

Green alga, Raphidocelis
subcapitata

(formerly, Pseudokirchneriella
subcapitata and Selenastnim
capriconnituni)

S, M

96 hr

Perfluorobutane

sulfonate
potassium salt /
97.3%

00

23.8-
24.5

ECso
(population
abundance)

2,146



Wildlife

International 2001

Green alga,

Raphidocelis subcapitata

S,U

48 hr

1 -Butanesulfonic

acid,
1,1,2,2,3,3,4,4,4-

Nonafluoro-,
Potassium salt /
>95%

7.2

20

ECio

(population
growth rate)

299

Test duration too
short

Kusk et al. 2018



Zebrafish (embryo),
Danio retio

S,U

144 hr
post fert.

Perfluorobutane
sulfonic acid / NR

7.2-

7.6

26

LCso

1,500

Duration too long
for an acute test and
too short for a
chronic test

Ulhaqetal. 2013

Zebrafish (embryo, 6 hpf),
Danio retio

S,U

114 hr

Potassium
nonafluoro-1-
butanesulfonate /
NR

NR

28

NOEC

(growtli

stunting)

20.84b

Duration too long
for an acute test and
too short for a
chronic test

Truong et al. 2014

Zebrafish (embryo),
Danio retio

R,U

168 hr
post fert.

Nonafluorobutane-
1-sulfonic acid /
NR

7.5

28-
28.5

LCso

921.9

Duration too long
for an acute test and
too short for a
chronic test

Stinckens et al.
2018

Zebrafish (embryo, 3 hpf),
Danio retio

R,U

5 d post
fert.

Nonafluorobutane-
1-sulfonic acid /
97%

NR

NR

LCso

406.2b

Test duration too
long for an acute
test and too short for
a chronic test

Sant et al. 2019

C-2


-------
Species (lifestage)

Method3

Test
Duration

Chemical /
Purity

pH

Temp.
(°C)

Effect

Author
Reported
Effect
Concentration
(mg/L)

Deficiencies

Reference

Zebrafish (embryo, 6 hpf),
Danio retio

S,U

114 hr

Potassium
nonafluoro-1-
butanesulfonate /
NR

NR

NR

BMDio
(mortality)
(benchmark
dose)

27.75b

Test duration too
long for an acute
test and too short for
a chronic test

Thomas et al. 2019

Zebrafish (embryo, 5-6 hpf),
Danio retio

R,U

90-91 hr

Potassium
perfluorobutane-
sulfonate / >99%

NR

28

LCso

1,394

Test duration too
short

Wasel et al. 2020

Zebrafish (embryo, 6-8 hpf),
Danio retio

S,U

112-114
hr

Potassium
perfluoro-1-
butanesulfonate /

>98%

NR

28

NOEC
(mortality)

0.2295b

Only one
concentration
tested; duration too
long for an acute
test and too short for
a chronic test

Rericha et al. 2021

Zebrafish (3-yr old adults),
Danio retio

R,U

14 d

Perfluorobutanesul
fonate /

>98%

NR

NR

LOEC
(multiple
genetic and
biochemistry
endpoints)

0.100

Only non-apical
endpoints evaluated

Liu et al. 2022

Perfluorohexanoic acid (PFHxA)

Rainbow trout (oocyte, ova),
Oncorhvnchiis mykiss

S, M

3 hr

Perfluorohexanoic
acid / >97%

8.5

6

NOEC
(accumulation
residue)

10.83

Test duration too
short

Raine et al. 2021



Zebrafish (embryo, 6 hpf),
Danio retio

S,U

114 hr

Perfluorohexanoic
acid / NR

NR

28

NOEC

(growth

stunting)

20.10b

Duration too long
for an acute test and
too short for a
chronic test

Truong et al. 2014

Zebrafish (embryo, 6 hpf),
Danio retio

S,U

114 hr

Perfluorohexanoic
acid / NR

NR

NR

BMDio
(mortality)
(benchmark
dose)

22.18b

Duration too long
for an acute test and
too short for a
chronic test

Thomas et al. 2019

Zebrafish (embryo, 5-6 hpf),
Danio retio

R, U

90-91 hr

Perfluorohexanoic
acid / >99%

7

28

LCso

6,486

Test duration too
short

Wasel et al. 2020

Zebrafish (embryo, 6-8 hpf),
Danio retio

S,U

120 hr

Perfluoro-n-
hexanoic acid /

>98%

NR

28

MATC
(mortality)

5.367

Duration too long
for an acute test and
too short for a
chronic test

Guo et al. 2021

C-3


-------
Species (lifestage)

Method3

Test
Duration

Chemical /
Purity

pH

Temp.
(°C)

Effect

Author
Reported
Effect
Concentration
(mg/L)

Deficiencies

Reference

Zebrafish (embryo, 6-8 hpf),
Danio retio

S,U

112-114
hr

Perfluoro-n-
hexanoic acid /

>98%

NR

28

NOEC
(mortality)

0.2512b

Duration too long
for an acute test and
too short for a
chronic test

Rericha et al. 2021

Zebrafish (embryo, 0.5-2 hpf),
Danio retio

R,U

94-95.5 hr

Perfluorohexanoic
acid / >99%

NR

28

MATC
(mortality)

5.367

Test duration too
short

Zhang et al. 2022

Perfluorohexanesulfonic acid (PFHxS)

Zebrafish (embryo, 6 hpf),
Danio retio

S,U

114 hr

Potassium
perfluorohexanesul
fonate / NR

NR

28

MATC
(mortality)

8.889b

Duration too long
for an acute test and
too short for a
chronic test

Truong et al. 2014

Zebrafish (embryo, 6 hpf),
Danio retio

S,U

114 lir

Potassium
perfluorohexanesul
fonate / NR

NR

NR

BMDio
(mortality)
(benchmark
dose)

1.968b

Duration too long
for an acute test and
too short for a
chronic test

Thomas et al. 2019

Zebrafish (embryo, 6-8 hpf),
Danio retio

s,u

112-114
hr

Sodium perfluoro-
1 -hexanesulfonate

/ >98%

NR

28

NOEC
(mortality)

0.2503b

Duration too long
for an acute test and
too short for a
chronic test

Rericha et al. 2021

Perfluorononanoic acid (PFNA)

Green alga, Raphidocelis
subcapitata

(formerly, Pseudokirchneriella
subcapitata and Selenastnim
capriconnituni)

s,u

4.5 hr

Perfluorononanoic
acid/97%

NR

NR

ECso
(population
photosyntliesis)

481.7b

Test duration too
short

Ding et al. 2012



Duckweed,
Lemna gibba

s,u

7 d

Perfluorononanoic
acid / >97%

NR

25

IC50

(population/
weight)

89.10b

Test duration too
long

Boudreau 2002



Cladoceran (<24 h old),
Daphnia pulicaria

R,U

21 d

Perfluorononanoic
acid / >97%

NR

21

MATC
(reproduction:
young/brood)

8.694b

Lack of procedural
details cannot judge
against data quality
objectives

Boudreau 2002



Zebrafish (embryo, 0 hpf),
Danio retio

S,U

72 lir post
fert.

Perfluorononanoic
acid/97%

8.3

28.5

LCso

84

Test duration too
short

Zheng et al. 2012

C-4


-------
Species (lifestage)

Method3

Test
Duration

Chemical /
Purity

pH

Temp.
(°C)

Effect

Author
Reported
Effect
Concentration
(mg/L)

Deficiencies

Reference

Zebrafish (embryo),
Danio retio

S,U

144 hr
post fert.

Perfluorononanoic
acid / NR

7.2-

7.6

26

LCso

>10

Duration too long
for an acute test and
too short for a
chronic test

Ulhaqetal. 2013

Zebrafish (embryo, 6 hpf),
Danio retio

s,u

114 hr

Perfluorononanoic
acid / NR

NR

28

MATC
(mortality)

0.9392b

Duration too long
for an acute test and
too short for a
chronic test

Truong et al. 2014

Zebrafish (embryo, 1 hpf),
Danio retio

s,u

23 hr

Perfluorononanoic
acid/97%

7.2-

8.0

28

LCso

140.2b

Test duration too
short

Liu et al. 2015

Zebrafish (embryo, 3 hpf),
Danio retio

s,u

117 hr

Perfluorononanoic
acid / NR

7.2-

7.7

26-28

MATC
(growth -
length)

0.2935b

Duration too long
for an acute test and
too short for a
chronic test

Jantzen et al. 2016

Zebrafish (embryo, 72 hpf),
Danio retio

S, M

48 hr

Perfluorononanoic
acid / NR

NR

27

LCso

108.6

Test duration too
short

Rainieri et al. 2017

Zebrafish (embryo, 6 hpf),
Danio retio

S,U

114 hr

Perfluorononanoic
acid / NR

NR

NR

BMDio
(mortality)
(benchmark
dose)

17.10b

Duration too long
for an acute test and
too short for a
chronic test

Thomas et al. 2019

Zebrafish (embryo, 6-8 hpf),
Danio retio

s,u

112-114
hr

Perfluoro-n-
nonanoic acid /
97%

NR

28

LOEC
(development
deformation)

45.02b

Duration too long
for an acute test and
too short for a
chronic test

Rericha et al. 2021



African clawed frog (embryo, 8-
11 stage),

Xenopiis laevis

R,U

96 hr

Perfluorononanoic
acid / NR

6.5-

7.5

24

NOEC
(mortality,
growth)

3.5

Only one

concentration

evaluated

Mitchell 2010

Perfluorodecanoic acid (PFDA)

Green alga, Raphidocelis
subcapitata

(formerly, Pseudokirchneriella
subcapitata and Selenastnim
capriconmtum)

S,U

4.5 hr

Perfluorodecanoic
acid/

98%

NR

NR

ECso
(population
photosynthesis)

437.5b

Test duration too
short

Ding et al. 2012

Green alga,

Raphidocelis subcapitata

s,u

72 hr

n-

Perfluorodecanoic
acid/
99.6%

-7.5

24

ECso
(population
biomass)

10.6

Test duration too
short

Hoke et al. 2012

C-5


-------
Species (lifestage)

Method3

Test
Duration

Chemical /
Purity

PH

Temp.
(°C)

Effect

Author
Reported
Effect
Concentration
(mg/L)

Deficiencies

Reference



Duckweed,
Lemna gibbet

S,U

7 d

Perfluorodecanoic
acid/

>97%

NR

25

IC50

(population/
weight)

99.22b

Test duration too
long

Boudreau 2002



Zebrafish (embryo),
Danio retio

s,u

144 hr
post fert.

Perfluorodecanoic
acid / NR

7.2-

7.6

26

LCso

8.4

Duration too long
for an acute test and
too short for a
chronic test

Ulhaqetal. 2013

Zebrafish (embryo, 6 hpf),
Danio retio

s,u

114 hr

Perfluorodecanoic
acid / NR

NR

28

NOEC

(growth

stunting)

32.90b

Duration too long
for an acute test and
too short for a
chronic test

Truong et al. 2014

Zebrafish (embryo, 6 hpf),
Danio retio

s,u

114 hr

Perfluorodecanoic
acid / NR

NR

NR

BMDio
(mortality)
(benclimark
dose)

5,141b

Duration too long
for an acute test and
too short for a
chronic test

Thomas et al. 2019

Zebrafish (embryo, 6-8 hpf),
Danio retio

s,u

112-114
hr

Perfluoro-n-
decanoic acid /
98%

NR

28

NOEC
(mortality)

50.38b

Only one
concentration
tested; duration too
long for an acute
test and too short for
a chronic test

Rericha et al. 2021



African clawed frog (embryo, 8-
11 stage),

Xenopiis laevis

R,U

96 hr

Perfluorodecanoic
acid / NR

6.5-

7.5

24

NOEC
(mortality,
growth)

3.5

Only one

concentration

evaluated

Mitchell 2010

Hexadecafluoro-2-decenoic acid (8:2 FTUCA)

Green alga,

Raphidocelis subcapitata
(formerly, Pseudokirchneriella
subcapitata and Selenastnim
capriconnituni)

S, M

72 hr

8:2 Fluorotelomer
unsaturated
carboxylate /
>98%

6.5-

7.5

24

ECio

(population
abundance)

2.3

Test duration too
short

Mitchell et al. 2011

Green alga, Raphidocelis
subcapitata

S,U

72 hr

8:2 FTUCA /
99.8%

-7.5

24

ECso
(population
biomass)

7.5

Test duration too
short

Hoke et al. 2012



C-6


-------
Species (lifestage)

Method3

Test
Duration

Chemical /
Purity

pH

Temp.
(°C)

Effect

Author
Reported
Effect
Concentration
(mg/L)

Deficiencies

Reference

Green alga (exponential growth
phase (log), 3-7 d),

Chlorella vulgaris

S, M

72 hr

8:2 Fluorotelomer
unsaturated
carboxylate /
>98%

6.5-

7.5

24

ECio

(population
abundance)

2.2

Test duration too
short

Mitchell et al. 2011



Amphipod,
Hvalella azteca

R,M

10 d

8:2 Fluorotelomer
unsaturated
carboxylate /
>98%

6.5-

7.5

23

NOEC
(mortality and
growth)

6.1

Duration too long
for an acute test and
too short for a
chronic test

Mitchell et al. 2011



Zebrafish (embryo, 6-8 hpf),
Danio retio

S,U

112-114
hr

2H-Perfluoro-2-
decenoic acid /

>98%

NR

28

NOEC
(mortality)

0.2520b

Only one
concentration
tested; duration too
long for an acute
test and too short for
a chronic test

Rericha et al. 2021

Pentadecafluorodecanoic acid (7:3 FTCA)

Green alga, Raphidocelis
subcapitata

(formerly, Pseudokirchneriella
subcapitata and Selenastrum
capriconmtum)

S,U

72 hr

4,4,5,5,6,6,7,7,8,8,

9,9,10,10,10-
Pentadecafluorode
canoic acid / >99%

-7.5

24

ECso
(population
biomass)

2.1

Test duration too
short

Hoke et al. 2012



Zebrafish (embryo, 6-8 hpf),
Danio retio

S,U

112-114
hr

3-Perfluoroheptyl
propanoic acid /

>98%

NR

28

NOEC
(mortality)

0.2520b

Only one
concentration
tested; duration too
long for an acute
test and too short for
a chronic test

Rericha et al. 2021

a S=static, R=renewal, F=flow-through, U=unmeasured, M=measurec.
b Converted to mg/L based on molecular weight.

C-7


-------
C.l References

Perfluorobutanoic acid (PFBA)

Boudreau, T.M. 2002. Toxicity of perfluorinated organic acids to selected freshwater organisms
under laboratory and field conditions. M.S. Thesis, University of Guelph, Ontario, Canada: 145 p.

Ding, G., M. Wouterse, R. Baerselman and W.J.G.M. Peijnenburg. 2012. Toxicity of
polyfluorinated and perfluorinated compounds to lettuce {Lactuca sativa) and green algae
(Pseudokirchneriella subcapitata). Arch. Environ. Contam. Toxicol. 62(1): 49-55.

Godfrey, A.E.. 2017. Endocrine disrupting effects of halogenated chemicals on fish. Ph.D.
Thesis, Purdue University, West Lafayette, IN: 125 p.

Godfrey, A., B. Hooser, A. Abdelmoneim, and M.S. Sepulveda. 2019. Sex-Specific Endocrine-
Disrupting Effects of Three Halogenated Godfrey,A., B. Hooser, A. Abdelmoneim, and M.S.
Sepulveda. 2019. Sex-Specific Endocrine-Disrupting Effects of Three Halogenated Chemicals in
Japanese Medaka. J. Appl. Toxicol. 39(8): 1215-1223.

Rericha, Y., D. Cao, L. Truong, M. Simonich, J.A. Field and R.L. Tanguay. 2021. Behavior
effects of structurally diverse per- and polyfluoroalkyl substances in zebrafish. Chem. Res.
Toxicol. 34(6): 1409-1416.

Ulhaq, M., G. Carlsson, S. Orn and L. Norrgren. 2013. Comparison of developmental toxicity of
seven perfluoroalkyl acids to zebrafish embryos. Environ. Toxicol. Pharmacol. 36: 423-426.

Wasel, O., K.M. Thompson, Y. Gao, A.E. Godfrey, J. Gao, C.T. Mahapatra, L.S. Lee, M.S.
Sepulveda and J.L. Freeman. 2020. Comparison of zebrafish in vitro and in vivo developmental
toxicity assessments of perfluoroalkyl acids (PFAAs). J. Toxicol. Environ. Health Part A. 84:
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Perfluorobutanesulfonic acid (PFBS)

Kusk, K.O., A.M. Christensen and N. Nyholm. 2018. Algal growth inhibition test results of 425
organic chemical substances. Chemosphere. 204: 405-412.

Liu, M.Y., B.L. Sun, X.Z. Zhou and L.G. Chen. 2022. Disturbed glucose metabolism by
perfluorobutanesulfonate pollutant and benefit of young fecal transplantation in aged zebrafish.
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Rericha, Y., D. Cao, L. Truong, M. Simonich, J.A. Field and R.L. Tanguay. 2021. Behavior
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Sant, K.E., O.L. Venezia, P.P. Sinno and A.R. Timme-Laragy. 2019. Perfluorobutanesulfonic
acid disrupts pancreatic organogenesis and regulation of lipid metabolism in the zebrafish, Danio
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Stinckens, E., L. Vergauwen, G.T. Ankley, R. Blust, V.M. Darras, D.L. Villeneuve, H. Witters,
D.C. Volz and D. Knapen. 2018. An AOP-based alternative testing strategy to predict the impact
of thyroid hormone disruption on swim bladder inflation in zebrafish. Aquat. Toxicol. 200: 1-12.

Thomas, D.G., H. Shankaran, L. Truong, R.L. Tanguay andK.M. Waters. 2019. Time-dependent
behavioral data from zebrafish reveals novel signatures of chemical toxicity using point of
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Truong, L., D.M. Reif, L. St Mary, M.C. Geier, H.D. Truong and R.L. Tanguay. 2014.
Multidimensional in vivo hazard assessment using zebrafish. Toxicol. Sci. 137(1): 212-233.

Ulhaq, M., G. Carlsson, S. Orn and L. Norrgren. 2013. Comparison of developmental toxicity of
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Wasel, O., K.M. Thompson, Y. Gao, A.E. Godfrey, J. Gao, C.T. Mahapatra, L.S. Lee, M.S.
Sepulveda and J.L. Freeman. 2020. Comparison of zebrafish in vitro and in vivo developmental
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Guo, X., S. Zhang, X. Liu, S. Lu, Q. Wu and P. Xie. 2021. Evaluation of the acute toxicity and
neurodevelopmental inhibition of perfluorohexanoic acid (PFHxA) in zebrafish embryos.
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Raine, J.C., S. Su, E. Lin, Z.L. Yang, J.P. Giesy and P.D. Jones. 2021. Prefertilization exposure
of rainbow trout eggs to per- and polyfluoroalkyl substances to simulate accumulation during
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Rericha, Y., D. Cao, L. Truong, M. Simonich, J.A. Field and R.L. Tanguay. 2021. Behavior
effects of structurally diverse per- and polyfluoroalkyl substances in zebrafish. Chem. Res.
Toxicol. 34(6): 1409-1416.

Thomas, D.G., H. Shankaran, L. Truong, R.L. Tanguay andK.M. Waters. 2019. Time-dependent
behavioral data from zebrafish reveals novel signatures of chemical toxicity using point of
departure analysis. Comput. Toxicol. 9: 50-60.

Truong, L., D.M. Reif, L. St Mary, M.C. Geier, H.D. Truong and R.L. Tanguay. 2014.
Multidimensional in vivo hazard assessment using zebrafish. Toxicol. Sci. 137(1): 212-233.

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Wasel, O., K.M. Thompson, Y. Gao, A.E. Godfrey, J. Gao, C.T. Mahapatra, L.S. Lee, M.S.
Sepulveda and J.L. Freeman. 2020. Comparison of zebrafish in vitro and in vivo developmental
toxicity assessments of perfluoroalkyl acids (PFAAs). J. Toxicol. Environ. Health Part A. 84:
125-136.

Zhang, S.N., X. Guo, S. Lu, J. He, Q. Wu, X. Liu, Z.Y. Han and P. Xie. 2022. Perfluorohexanoic
acid caused disruption of the hypothalamus-pituitary-thyroid axis in zebrafish larvae. Ecotoxicol.
Environ. Saf. 2022 Mar 1:232: 113283.

Perfluorohexanesulfonic acid (PFHxS)

Rericha, Y., D. Cao, L. Truong, M. Simonich, J.A. Field and R.L. Tanguay. 2021. Behavior
effects of structurally diverse per- and polyfluoroalkyl substances in zebrafish. Chem. Res.
Toxicol. 34(6): 1409-1416.

Thomas, D.G., H. Shankaran, L. Truong, R.L. Tanguay and K.M. Waters. 2019. Time-dependent
behavioral data from zebrafish reveals novel signatures of chemical toxicity using point of
departure analysis. Comput. Toxicol. 9: 50-60.

Truong, L., D.M. Reif, L. St Mary, M.C. Geier, H.D. Truong and R.L. Tanguay. 2014.
Multidimensional in vivo hazard assessment using zebrafish. Toxicol. Sci. 137(1): 212-233.

Perfluorononanoic acid (PFNA)

Boudreau, T.M. 2002. Toxicity of perfluorinated organic acids to selected freshwater organisms
under laboratory and field conditions. M.S. Thesis, University of Guelph, Ontario, Canada: 145 p.

Ding, G., M. Wouterse, R. Baerselman and W.J.G.M. Peijnenburg. 2012. Toxicity of
polyfluorinated and perfluorinated compounds to lettuce (Lactuca sativa) and green algae
(Pseudokirchneriella subcapitata). Arch. Environ. Contam. Toxicol. 62(1): 49-55.

Jantzen, C.E., K.A. Annunziato, S.M. Bugel and K.R. Cooper. 2016. PFOS, PFNA, and PFOA
sub-lethal exposure to embryonic zebrafish have different toxicity profiles in terms of
morphometries, behavior and gene expression. Aquat. Toxicol. 175: 160-170.

Liu, H., N. Sheng, W. Zhang and J. Dai. 2015. Toxic effects of perfluorononanoic acid on the
development of zebrafish (Danio rerio) embryos. J. Environ. Sci. (China). 32: 26-34.

Mitchell, R.J. 2010. The toxicity of fluorotelomer acids to freshwater organisms and a
preliminary evaluation of mechanism of action. M.S. Thesis, University of Guelph, Ontario,
Canada: 118 p.

Rainieri, S., N. Conlledo, T. Langerholc, E. Madorran, M. Sala and A. Barranco. 2017. Toxic
effects of perfluorinated compounds at human cellular level and on a model vertebrate. Food
Chem. Toxicol. 104: 14-25.

C-10


-------
Rericha, Y., D. Cao, L. Truong, M. Simonich, J.A. Field and R.L. Tanguay. 2021. Behavior
effects of structurally diverse per- and polyfluoroalkyl substances in zebrafish. Chem. Res.
Toxicol. 34(6): 1409-1416.

Thomas, D.G., H. Shankaran, L. Truong, R.L. Tanguay andK.M. Waters. 2019. Time-dependent
behavioral data from zebrafish reveals novel signatures of chemical toxicity using point of
departure analysis. Comput. Toxicol. 9: 50-60.

Truong, L., D.M. Reif, L. St Mary, M.C. Geier, H.D. Truong and R.L. Tanguay. 2014.
Multidimensional in vivo hazard assessment using zebrafish. Toxicol. Sci. 137(1): 212-233.

Ulhaq, M., G. Carlsson, S. Orn and L. Norrgren. 2013. Comparison of developmental toxicity of
seven perfluoroalkyl acids to zebrafish embryos. Environ. Toxicol. Pharmacol. 36: 423-426.

Zheng, X.M., H.L. Liu, W. Shi, S. Wei, J.P. Giesy and H.X. Yu. 2012. Effects of perfluorinated
compounds on development of zebrafish embryos. Environ. Sci. Pollut. Res. 19(7): 2498-2505.

Perfluorodecanoic acid (PFDA)

Boudreau, T.M. 2002. Toxicity of perfluorinated organic acids to selected freshwater organisms
under laboratory and field conditions. M.S. Thesis, University of Guelph, Ontario, Canada: 145
P-

Ding, G., M. Wouterse, R. Baerselman and W.J.G.M. Peijnenburg. 2012. Toxicity of
polyfluorinated and perfluorinated compounds to lettuce (Lactuca sativa) and green algae
(Pseudokirchneriella subcapitata). Arch. Environ. Contam. Toxicol. 62(1): 49-55.

Hoke, R.A., L.D. Bouchelle, B.D. Ferrell and R.C. Buck. 2012. Comparative acute freshwater
hazard assessment and preliminary PNEC development for eight fluorinated acids.

Chemosphere. 87(7): 725-733.

Mitchell, R.J. 2010. The toxicity of fluorotelomer acids to freshwater organisms and a
preliminary evaluation of mechanism of action. M.S. Thesis, University of Guelph, Ontario,
Canada: 118 p.

Rericha, Y., D. Cao, L. Truong, M. Simonich, J.A. Field, and R.L. Tanguay. 2021. Behavior
effects of structurally diverse per- and polyfluoroalkyl substances in zebrafish. Chem. Res.
Toxicol. 34(6): 1409-1416.

Thomas, D.G., H. Shankaran, L. Truong, R.L. Tanguay andK.M. Waters. 2019. Time-dependent
behavioral data from zebrafish reveals novel signatures of chemical toxicity using point of
departure analysis. Comput. Toxicol. 9:50-60.

Truong, L., D.M. Reif, L. St Mary, M.C. Geier, H.D. Truong, and R.L. Tanguay. 2014.
Multidimensional in vivo hazard assessment using zebrafish. Toxicol. Sci. 137(1): 212-233.

C-ll


-------
Ulhaq, M., G. Carlsson, S. Orn, and L. Norrgren. 2013. Comparison of developmental toxicity of
seven perfluoroalkyl acids to zebrafish embryos. Environ. Toxicol. Pharmacol. 36: 423-426.

Hexadecafluoro-2-decenoic acid (8:2 FTUCA)

Hoke, R.A., L.D. Bouchelle, B.D. Ferrell and R.C. Buck. 2012. Comparative acute freshwater
hazard assessment and preliminary PNEC development for eight fluorinated acids.

Chemosphere. 87(7): 725-733.

Mitchell, R.J., A.L. Myers, S.A. Mabury, K.R. Solomon and P.K. Sibley. 2011. Toxicity of
fluorotelomer carboxylic acids to the algae Pseudokirchneriella subcapitata and Chlorella
vulgaris, and the amphipod Hyalella azteca. Ecotoxicol. Environ. Saf. 74(8): 2260-2267.

Rericha, Y., D. Cao, L. Truong, M. Simonich, J.A. Field and R.L. Tanguay. 2021. Behavior
effects of structurally diverse per- and polyfluoroalkyl substances in zebrafish. Chem. Res.
Toxicol. 34(6): 1409-1416.

Pentadecafluorodecanoic acid (7:3 FTCA)

Hoke, R.A., L.D. Bouchelle, B.D. Ferrell and R.C. Buck. 2012. Comparative acute freshwater
hazard assessment and preliminary PNEC development for eight fluorinated acids.

Chemosphere. 87(7): 725-733.

Rericha, Y., D. Cao, L. Truong, M. Simonich, J.A. Field and R.L. Tanguay. 2021. Behavior
effects of structurally diverse per- and polyfluoroalkyl substances in zebrafish. Chem. Res.
Toxicol. 34(6): 1409-1416.

C-12


-------
Appendix D Unused Freshwater Acute Toxicity Studies

Author

Citation

Reason Unused

Perfluorobutanoic acid (PFBA)

Blanc, M., J. Ruegg, N.
Scherbak and S.H. Keiter

2019. Environmental chemicals differentially affect
epigenetic-related mechanisms in the zebrafish liver
(ZF-L) cell line and in zebrafish embryos. Aquat.
Toxicol. 215: 105272-9999.

No control
information

Perfluorononanoic acid (PFNA)

Dale, K., F. Yadetie, T.
Horvli, X. Zhang, H.G.
Froysa, OA. Karlsen and A.
Goksoyr

2022. Single PFAS and PFAS mixtures affect
nuclear receptor- and oxidative stress-related
pathways in precision-cut liver slices of Atlantic cod
(Gctdus morhaa). Sci. Total Environ. 814: 1-12.

In vitro exposure

Garoche, C., A. Boulahtouf,
M. Grimaldi, B. Chiavarina,
L. Toporova, M.J. Den
Breeder, J. Legler, W.
Bourguet and P. Ba

2021. Interspecies differences in activation of
peroxisome proliferator-activated receptor gamma
by pharmaceutical and environmental chemicals.
Environ. Sci. Technol. 55(24): 16489-16501.

In vitro exposure

D-l


-------
Appendix E ICE Models Available in Web-ICE v4.0 for Freshwater Predicted Species Based on
Surrogates with Measured Toxicities

Table E-l. All ICE Models Selected for the Derivation of PFAS Benchmarks.

Model parameters are used to evaluate prediction robustness. Cross-validation success is the percentage of all model data that were predicted within five-fold of
the measured value through leave-one-out cross-validation (Raimondo et al. 2024). Taxonomic distance describes the relationship between surrogate and predicted

Predicted Species

Surrogate Species

Slope

Intercept

Degrees
of

Freedom
(N-2)

R2

p-value

Mean
Square
Error
(MSE)

Surrogate

model
minimum
value
(Hg/L)

Surrogate

model
maximum
value (ng/L)

Cross-
validation
Success
(%)

Taxonomic
Distance

PFAS
Compound

African clawed frog
(Xenopus laevis)

Fathead minnow

(Pimephales

promelas)

0.76

0.77

4

0.91

0.00

0.14

18.0

140,225.3

100

5

PFBS

Amphipod

(Crangonyx

pseudogracilis)

Daphnid (Daphnia
magna)

0.91

1.26

12

0.75

0.00

0.76

0.8

166,819.4

50

5

PFNA, PFHxA,
PFDA, PFBS,
PFBA, 7:3
FTCA, 8:2
FTUCA

Amphipod

(Gammaras

fasciatus)

Daphnid (Daphnia
magna)

0.82

0.44

51

0.75

0.00

0.76

0.2

5,000,000.0

49

5

PFNA, PFHxA,
PFDA, PFBS,
PFBA, 7:3
FTCA, 8:2
FTUCA

Amphipod

(Gammaras

fasciatus)

Mysid

(Americamysis
bahia)

0.91

0.58

26

0.70

0.00

0.81

0.0

4,400.0

57

4

PFBS

Amphipod
(Gammaras minus)

Fathead minnow

(Pimephales

promelas)

0.73

1.50

2

0.96

0.02

0.05

239.0

107,584.8

75

6

PFBS

Amphipod

(Gammaras

pseudolimnaeus)

Daphnid (Daphnia
magna)

0.92

0.25

30

0.75

0.00

0.75

0.1

68,300.0

63

5

PFNA, PFHxA,
PFDA, PFBS,
PFBA, 7:3
FTCA, 8:2
FTUCA

Amphipod

(Gammaras

pseudolimnaeus)

Mysid

(Americamysis
bahia)

0.82

0.22

18

0.80

0.00

0.56

0.0

7,170.0

55

4

PFBS

Amphipod (Hyalella
azteca)

Mysid

(Americamysis
bahia)

1.03

-0.10

30

0.88

0.00

0.68

0.0

193,390.8

63

4

PFBS

E-l


-------
Predicted Species

Surrogate Species

Slope

Intercept

Degrees
of

Freedom
(N-2)

R2

p-value

Mean
Square
Error
(MSE)

Surrogate

model
minimum
value
(Hg/L)

Surrogate

model
maximum
value (ng/L)

Cross-
validation
Success
(%)

Taxonomic
Distance

PFAS
Compound

Apache trout
(Oncorhynchus gilae)

Rainbow trout

(Oncorhynchus

mykiss)

1.10

-0.35

3

0.99

0.00

0.02

4.1

1,624.7

100

1

PFDA, 7:3
FTCA, 8:2
FTUCA

Arctic grayling
(Thymallus arcticus)

Bluegill (Lepomis
macrochirus)

1.14

-0.59

1

1.00

0.02

0.01

37.9

150,750.0

N/A

4

PFBS

Atlantic salmon
(Salmo salar)

Bluegill (Lepomis
macrochirus)

1.15

-0.64

11

0.95

0.00

0.13

4.3

123,267.3

92

4

PFBS

Atlantic salmon
(Salmo salar)

Rainbow trout

(Oncorhynchus

mykiss)

1.01

-0.07

12

0.96

0.00

0.11

0.2

95,857.7

86

2

PFDA, 7:3
FTCA, 8:2
FTUCA

Atlantic salmon
(Salmo salar)

Fathead minnow

(Pimephales

promelas)

1.28

-1.51

6

0.68

0.01

0.84

44.4

140,225.3

63

4

PFBS

Beaver-tail fairy
shrimp

(Thamnocephalus
platyurus)

Daphnid (Daphnia
magna)

0.91

0.22

18

0.97

0.00

0.12

0.3

8,694,505.9

95

4

PFNA, PFHxA,
PFDA, PFBS,
PFBA, 7:3
FTCA, 8:2
FTUCA

Black bullhead
(Ameiurns melas)

Rainbow trout

(Oncorhynchus

mykiss)

0.74

1.21

12

0.60

0.00

0.95

0.0

9,809.6

43

4

PFDA, 7:3
FTCA, 8:2
FTUCA

Black bullliead
(Ameiurus melas)

Fathead minnow

(Pimephales

promelas)

1.07

-0.10

3

0.97

0.00

0.17

1.2

20,922.0

80

4

PFBS

Bluegill (Lepomis
macrochirus)

Daphnid (Daphnia
magna)

0.71

1.18

270

0.65

0.00

0.84

0.1

46,278,306.5

57

6

PFNA, PFHxA,
PFDA, PFBS,
PFBA, 7:3
FTCA, 8:2
FTUCA

Bluegill (Lepomis
macrochirus)

Mysid

(Americamysis
bahia)

0.71

1.38

150

0.71

0.00

0.55

0.0

79,000.0

72

6

PFBS

Bluegill (Lepomis
macrochirus)

Rainbow trout

(Oncorhynchus

mykiss)

0.94

0.30

314

0.89

0.00

0.22

0.1

8,341,462.7

90

4

PFDA, 7:3
FTCA, 8:2
FTUCA

Bluegill (Lepomis
macrochirus)

Fathead minnow

(Pimephales

promelas)

0.94

-0.02

108

0.81

0.00

0.43

0.2

50,154,461.4

82

4

PFBS

E-2


-------
Predicted Species

Surrogate Species

Slope

Intercept

Degrees
of

Freedom
(N-2)

R2

p-value

Mean
Square
Error
(MSE)

Surrogate

model
minimum
value
(Hg/L)

Surrogate

model
maximum
value (ng/L)

Cross-
validation
Success
(%)

Taxonomic
Distance

PFAS
Compound

Bluegill (Lepomis
macrochiriis)

Bullfrog

(Lithobates

catesbeianus)

1.10

-0.48

5

0.98

0.00

0.10

2.5

233,000.0

100

5

PFHxS, PFHxA

Bonytail (Gila
elegans)

Bluegill (Lepomis
macrochiriis)

0.96

-0.37

6

0.69

0.01

0.62

53.9

150,750.0

50

4

PFBS

Bonytail (Gila
elegans)

Rainbow trout

(Oncorhynchus

mykiss)

0.92

0.70

6

0.89

0.00

0.22

3.3

43,073.0

75

4

PFDA, 7:3
FTCA, 8:2
FTUCA

Bonytail (Gila
elegans)

Fathead minnow

(Pimephales

promelas)

1.01

-0.39

5

0.62

0.04

0.50

58.3

8,320.3

43

2

PFBS

Brook trout
(Salvelinus
fontinalis)

Bluegill (Lepomis
macrochiriis)

0.97

-0.06

21

0.86

0.00

0.27

0.4

123,267.3

83

4

PFBS

Brook trout
(Salvelinus
fontinalis)

Rainbow trout

(Oncorhynchus

mykiss)

1.00

0.01

25

0.95

0.00

0.11

0.6

96,692.7

93

2

PFDA, 7:3
FTCA, 8:2
FTUCA

Brook trout
(Salvelinus
fontinalis)

Fatliead minnow

(Pimephales

promelas)

0.95

-0.43

17

0.70

0.00

0.75

1.2

140,225.3

68

4

PFBS

Brown trout (Salmo
tratta)

Bluegill (Lepomis
macrochiriis)

0.91

-0.10

16

0.73

0.00

0.68

2.3

123,267.3

78

4

PFBS

Brown trout (Salmo
tratta)

Rainbow trout

(Oncorhynchus

mykiss)

1.00

0.02

20

0.97

0.00

0.07

0.2

95,857.7

100

2

PFDA, 7:3
FTCA, 8:2
FTUCA

Bryozoan

(Pectinatella

magnifica)

Daphnid (Daphnia
magna)

0.87

1.14

2

0.98

0.01

0.01

14.1

231.7

100

6

PFNA, PFDA,
7:3 FTCA, 8:2
FTUCA

Bullfrog (Lithobates
catesbeianus)

Bluegill (Lepomis
macrochiriis)

0.89

0.48

5

0.98

0.00

0.08

0.4

201,000.0

100

5

PFBS

Bullfrog (Lithobates
catesbeianus)

Rainbow trout

(Oncorhynchus

mykiss)

0.88

0.82

9

0.95

0.00

0.31

0.6

13,400,000.0

73

5

PFDA, 7:3
FTCA, 8:2
FTUCA

Bullfrog (Lithobates
catesbeianus)

Fatliead minnow

(Pimephales

promelas)

0.97

0.15

7

0.98

0.00

0.10

1.2

26,500,000.0

100

5

PFBS

E-3


-------
Predicted Species

Surrogate Species

Slope

Intercept

Degrees
of

Freedom
(N-2)

R2

p-value

Mean
Square
Error
(MSE)

Surrogate

model
minimum
value
(Hg/L)

Surrogate

model
maximum
value (ng/L)

Cross-
validation
Success
(%)

Taxonomic
Distance

PFAS
Compound

Cape Fear shiner

(Notropis

mekistocholas)

Rainbow trout

(Oncorhynchus

mykiss)

1.17

-0.23

3

0.99

0.00

0.01

4.1

1,624.7

100

4

PFDA, 7:3
FTCA, 8:2
FTUCA

Catla (Gibelion catla)

Fathead minnow

(Pimephales

promelas)

1.09

-0.36

2

0.97

0.02

0.02

333.0

8,320.3

100

2

PFBS

Channel catfish
(Ictalurus punctatus)

Zebrafish-embryo
(Danio rerio-
embryo)

0.77

0.95

5

0.91

0.00

0.07

144.8

304,541.0

86

4

PFHxS, PFBA

Channel catfish
(Ictalurus punctatus)

Bluegill (Lepomis
macrochirus)

0.81

0.74

77

0.77

0.00

0.43

0.4

517,824.6

82

4

PFBS

Channel catfish
(Ictalurus punctatus)

Rainbow trout

(Oncorhynchus

mykiss)

0.82

0.82

98

0.76

0.00

0.50

0.0

13,400,000.0

76

4

PFDA, 7:3
FTCA, 8:2
FTUCA

Channel catfish
(Ictalurus punctatus)

Fathead minnow

(Pimephales

promelas)

0.98

0.06

52

0.87

0.00

0.30

1.2

26,500,000.0

85

4

PFBS

Channel catfish
(Ictalurus punctatus)

Bullfrog

(Lithobates

catesbeianus)

1.02

-0.28

7

0.99

0.00

0.08

2.5

20,900,000.0

100

5

PFHxS, PFHxA

Channel catfish
(Ictalurus punctatus)

African clawed
frog (Xenopus
laevis)

1.00

0.47

3

0.93

0.01

0.20

18.4

122,473.4

100

5

PFNA

Chinook salmon

(Oncorhynchus

tshawytscha)

Bluegill (Lepomis
macrochirus)

0.91

-0.10

13

0.65

0.00

0.95

4.1

150,750.0

67

4

PFBS

Chinook salmon

(Oncorhynchus

tshawytscha)

Rainbow trout

(Oncorhynchus

mykiss)

1.02

0.02

18

0.96

0.00

0.11

3.3

724,430.8

95

1

PFDA, 7:3
FTCA, 8:2
FTUCA

Chinook salmon

(Oncorhynchus

tshawytscha)

Fathead minnow

(Pimephales

promelas)

1.16

-1.22

9

0.71

0.00

0.82

10.5

140,225.3

45

4

PFBS

Coho salmon

(Oncorhynchus

kisutch)

Bluegill (Lepomis
macrochirus)

0.91

0.01

25

0.79

0.00

0.47

0.4

150,750.0

78

4

PFBS

Coho salmon

(Oncorhynchus

kisutch)

Rainbow trout

(Oncorhynchus

mykiss)

1.03

0.06

36

0.99

0.00

0.04

0.0

43,073.0

100

1

PFDA, 7:3
FTCA, 8:2
FTUCA

E-4


-------
Predicted Species

Surrogate Species

Slope

Intercept

Degrees
of

Freedom
(N-2)

R2

p-value

Mean
Square
Error
(MSE)

Surrogate

model
minimum
value
(Hg/L)

Surrogate

model
maximum
value (ng/L)

Cross-
validation
Success
(%)

Taxonomic
Distance

PFAS
Compound

Coho salmon

(Oncorhynchus

kisutch)

Fathead minnow

(Pimephales

promelas)

0.91

-0.23

19

0.65

0.00

0.69

1.2

20,922.0

67

4

PFBS

Colorado squawfish

(Ptychocheilus

lucius)

Rainbow trout

(Oncorhynchus

mykiss)

0.91

0.92

7

0.73

0.00

0.56

3.3

43,073.0

67

4

PFDA, 7:3
FTCA, 8:2
FTUCA

Colorado squawfish

(Ptychocheilus

lucius)

Fatliead minnow

(Pimephales

promelas)

1.07

-0.58

6

0.68

0.01

0.49

58.3

20,922.0

63

2

PFBS

Common carp
(Cyprinus carpio)

Zebrafish-embryo
(Danio rerio-
embryo)

0.83

0.45

5

0.66

0.03

0.37

86.0

34,190.0

71

2

PFBA

Common carp
(Cyprinus carpio)

Bluegill (Lepomis
macrochirus)

0.86

0.60

30

0.79

0.00

0.47

0.9

180,000.0

81

4

PFBS

Common carp
(Cyprinus carpio)

Rainbow trout

(Oncorhynchus

mykiss)

0.81

0.92

47

0.74

0.00

0.52

0.2

198,443.9

76

4

PFDA, 7:3
FTCA, 8:2
FTUCA

Common carp
(Cyprinus carpio)

Fatliead minnow

(Pimephales

promelas)

0.99

0.06

25

0.84

0.00

0.21

4.7

133,000.0

89

2

PFBS

Cutthroat trout
(Oncorhynchus
clarkii)

Bluegill (Lepomis
macrochirus)

0.88

0.42

32

0.79

0.00

0.36

0.4

180,000.0

71

4

PFBS

Cutthroat trout
(Oncorhynchus
clarkii)

Rainbow trout

(Oncorhynchus

mykiss)

0.95

0.15

36

0.96

0.00

0.08

0.0

198,443.9

97

1

PFDA, 7:3
FTCA, 8:2
FTUCA

Cutthroat trout
(Oncorhynchus
clarkii)

Fatliead minnow

(Pimephales

promelas)

0.99

-0.45

22

0.82

0.00

0.35

1.2

140,225.3

79

4

PFBS

Cuvier's foam froglet

(Physalaemus

cuvieri)

Rainbow trout

(Oncorhynchus

mykiss)

0.61

1.97

2

0.99

0.01

0.01

709.9

155,541.7

100

5

PFDA, 7:3
FTCA, 8:2
FTUCA

Daphnid

(Ceriodaphnia dubia)

Daphnid (Daplinia
magna)

1.03

-0.28

40

0.95

0.00

0.24

0.3

46,278,306.5

83

2

PFNA, PFHxA,
PFDA, PFBS,
PFBA, 7:3
FTCA, 8:2
FTUCA

E-5


-------
Predicted Species

Surrogate Species

Slope

Intercept

Degrees
of

Freedom
(N-2)

R2

p-value

Mean
Square
Error
(MSE)

Surrogate

model
minimum
value
(Hg/L)

Surrogate

model
maximum
value (ng/L)

Cross-
validation
Success
(%)

Taxonomic
Distance

PFAS
Compound

Daplinid

(Ceriodaplinia dubia)

Mysid

(Americamysis
bahia)

0.98

-0.21

25

0.84

0.00

0.77

0.0

4,100,000.0

52

5

PFBS

Daplinid

(Ceriodaplinia

reticulata)

Daplinid (Daplinia
magna)

0.87

-0.03

7

0.70

0.01

0.24

0.8

231.7

89

2

PFNA, PFDA,
8:2 FTUCA

Daplinid (Daphnia
galeata)

Daplinid (Daplinia
magna)

0.92

0.27

4

0.96

0.00

0.08

0.1

646.2

83

1

PFNA, PFDA,
7:3 FTCA, 8:2
FTUCA

























PFNA, PFHxA,

Daplinid (Daplinia
longispina)

Daplinid (Daplinia
magna)

1.22

-0.57

2

0.99

0.01

0.06

8.9

10,359.2

100

1

PFDA, PFBS,
PFBA, 7:3
FTCA, 8:2
FTUCA

Daplinid (Daplinia
magna)

Water flea
(Chydorus
sphaericus)

1.04

0.27

7

0.98

0.00

0.06

7.1

462,000.0

100

3

PFNA, PFDA

Daplinid (Daplinia
magna)

Zebrafish (Danio
rerio)

0.92

-0.31

11

0.71

0.00

0.74

3.1

36,910.0

54

6

PFHxS, PFBA

Daplinid (Daplinia
magna)

Daplinid (Daplinia
pulicaria)

0.88

0.51

4

0.94

0.00

0.19

9.3

237,367.4

83

1

PFDA

Daplinid (Daplinia
magna)

Mysid

(Americamysis
bahia)

0.82

0.83

246

0.73

0.00

0.76

0.0

11,000,000.0

70

5

PFBS

Daplinid (Daplinia
magna)

Bullfrog

(Litliobates

catesbeianus)

0.82

0.68

9

0.84

0.00

0.79

2.5

20,900,000.0

73

6

PFHxS, PFHxA

























PFNA, PFHxA,

Daplinid (Daplinia
pulex)

Daplinid (Daplinia
magna)

1.01

-0.17

40

0.96

0.00

0.15

0.2

4,894,739.3

95

1

PFDA, PFBS,
PFBA, 7:3
FTCA, 8:2
FTUCA

























PFNA, PFHxA,

Daplinid (Daplinia
pulicaria)

Daplinid (Daplinia
magna)

1.07

-0.32

4

0.94

0.00

0.23

14.1

281,641.4

83

1

PFDA, PFBS,
PFBA, 7:3
FTCA, 8:2
FTUCA

E-6


-------








Degrees
of





Mean
Square

Surrogate

model
minimum

Surrogate
model

Cross-
validation













Freedom





Error

value

maximum

Success

Taxonomic

PFAS

Predicted Species

Surrogate Species

Slope

Intercept

(N-2)

R2

p-value

(MSE)

(Hg/L)

value (ng/L)

(%)

Distance

Compound

























PFNA, PFHxA,

Daplinid

(Simocephalus

semilatus)

Daplinid (Daphnia
magna)

0.94

-0.02

21

0.86

0.00

0.28

0.2

7,200.0

87

2

PFDA, PFBS,
PFBA, 7:3
FTCA, 8:2
FTUCA

Daplinid

(Simocephalus

vetulus)

Daplinid (Daphnia
magna)

0.75

0.61

5

0.88

0.00

0.35

0.1

166,819.4

57

2

PFNA, PFHxA,
PFDA, PFBS,
7:3 FTCA, 8:2
FTUCA

Fairy shrimp

(Streptocephalus

riibricaudatus)

Zebrafish-embryo
(Danio rerio-
embryo)

0.60

2.45

2

0.95

0.03

0.06

4622.4

5,100,709.2

75

6

PFHxS

Fathead minnow

(Pimephales

promelas)

Zebrafish (Danio
rerio)

1.07

-0.25

4

0.97

0.00

0.05

18.0

25,076.0

100

2

PFHxS, PFBA

Fathead minnow

Zebrafish-embryo























(Pimephales

(Danio rerio-

1.00

-0.11

41

0.93

0.00

0.22

23.0

54,578,562.0

84

2

PFHxS, PFBA

promelas)

embryo)















































PFNA, PFHxA,

Fathead minnow
(Pimephales

Daplinid (Daphnia
magna)

0.69

1.42

236

0.64

0.00

0.76

0.2

46,500,000.0

66

6

PFDA, PFBS,
PFBA, 7:3

promelas)





















FTCA, 8:2
FTUCA

Fathead minnow

(Pimephales

promelas)

Bluegill (Lepomis
macrochiriis)

0.87

0.62

108

0.81

0.00

0.40

0.1

27,540,000.0

81

4

PFBS

Fathead minnow

Mysid























(Pimephales
promelas)

(Americamysis
bahia)

0.62

1.74

104

0.62

0.00

0.75

0.0

11,000,000.0

60

6

PFBS

Fathead minnow

Rainbow trout





















PFDA, 7:3

(Pimephales
promelas)

(Oncorhynchus
mykiss)

0.90

0.62

146

0.87

0.00

0.27

0.2

13,400,000.0

86

4

FTCA, 8:2
FTUCA

Fathead minnow

Bullfrog























(Pimephales

(Litliobates

1.01

-0.10

7

0.98

0.00

0.10

2.5

20,900,000.0

100

5

PFHxS, PFHxA

promelas)

catesbeianus)























Fathead minnow

African clawed























(Pimephales
promelas)

frog (Xenopus
laevis)

1.20

-0.62

4

0.91

0.00

0.22

44.7

122,473.4

67

5

PFNA

E-7


-------
Predicted Species

Surrogate Species

Slope

Intercept

Degrees
of

Freedom
(N-2)

R2

p-value

Mean
Square
Error
(MSE)

Surrogate

model
minimum
value
(Hg/L)

Surrogate

model
maximum
value (ng/L)

Cross-
validation
Success
(%)

Taxonomic
Distance

PFAS
Compound

Fatmucket (Lampsilis
siliquoidea)

Daphnid (Daphnia
magna)

0.90

0.24

17

0.94

0.00

0.19

14.1

8,694,505.9

79

6

PFNA, PFHxA,
PFDA, PFBS,
PFBA, 7:3
FTCA, 8:2
FTUCA

Fatmucket (Lampsilis
siliquoidea)

Fathead minnow

(Pimephales

promelas)

0.94

-0.40

13

0.65

0.00

0.93

13.4

10,830,770.1

47

6

PFBS

Flagfish (Jordanella
floridae)

Zebrafish (Danio
rerio)

1.22

-0.78

2

0.99

0.00

0.01

7797.4

649,307.3

100

4

PFHxS, PFBA

Flagfish (Jordanella
floridae)

Bluegill (Lepomis
macrochirus)

0.86

0.47

7

0.89

0.00

0.30

0.4

113,000.0

78

4

PFBS

Flagfish (Jordanella
floridae)

Fathead minnow

(Pimephales

promelas)

0.91

0.11

8

0.84

0.00

0.46

1.2

32,537.5

80

4

PFBS

Goldfish (Carassius
auratus)

Zebrafish-embryo
(Danio rerio-
embryo)

0.97

0.33

7

0.96

0.00

0.06

144.8

304,541.0

100

2

PFHxS, PFBA

Goldfish (Carassius
auratus)

Bluegill (Lepomis
macrochirus)

0.78

0.98

27

0.71

0.00

0.51

0.4

201,000.0

72

4

PFBS

Goldfish (Carassius
auratus)

Rainbow trout

(Oncorhynchus

mykiss)

0.82

1.12

35

0.83

0.00

0.49

0.0

13,400,000.0

81

4

PFDA, 7:3
FTCA, 8:2
FTUCA

Goldfish (Carassius
auratus)

Fathead minnow

(Pimephales

promelas)

0.98

0.19

26

0.94

0.00

0.14

1.2

26,500,000.0

93

2

PFBS

Goldfish (Carassius
auratus)

Bullfrog

(Lithobates

catesbeianus)

1.04

-0.30

7

0.98

0.00

0.14

2.5

20,900,000.0

89

5

PFHxS, PFHxA

Green floater

(Lasmigona

subviridis)

Daplinid (Daplinia
magna)

0.67

0.94

2

0.97

0.02

0.08

14.1

166,819.4

75

6

PFNA, PFHxA,
PFDA, PFBS,
PFBA, 7:3
FTCA, 8:2
FTUCA

Green floater

(Lasmigona

subviridis)

Rainbow trout

(Oncorhynchus

mykiss)

0.60

1.44

4

0.73

0.03

0.36

3.3

95,857.7

67

6

PFDA

Green sunfish
(Lepomis cyanellus)

Bluegill (Lepomis
macrochirus)

0.85

0.63

13

0.92

0.00

0.14

4.1

113,000.0

93

1

PFBS

E-8


-------
Predicted Species

Surrogate Species

Slope

Intercept

Degrees
of

Freedom
(N-2)

R2

p-value

Mean
Square
Error
(MSE)

Surrogate

model
minimum
value
(Hg/L)

Surrogate

model
maximum
value (ng/L)

Cross-
validation
Success
(%)

Taxonomic
Distance

PFAS
Compound

Green simfish
(Lepomis cyaiiellus)

Rainbow trout

(Oncorhynchus

mykiss)

0.90

0.60

13

0.95

0.00

0.13

0.0

19,926.0

100

4

PFDA, 7:3
FTCA, 8:2
FTUCA

Green simfish
(Lepomis cyanellus)

Fathead minnow

(Pimephales

promelas)

0.90

-0.09

6

0.64

0.02

0.89

10.5

26,685.4

50

4

PFBS

Greentliroat darter
(Etheo stoma
lepidum)

Rainbow trout

(Oncorhynchus

mykiss)

1.05

-0.04

3

0.93

0.01

0.11

4.1

1,624.7

100

4

PFDA, 7:3
FTCA, 8:2
FTUCA

Greentliroat darter
(Etheo stoma
lepidum)

Fatliead minnow

(Pimephales

promelas)

0.85

-0.09

3

0.92

0.01

0.13

5.7

8,320.3

100

4

PFBS

Guppy (Poecilia
reticulata)

Bluegill (Lepomis
macrochirus)

0.70

1.04

22

0.75

0.00

0.39

0.4

180,000.0

67

4

PFBS

Guppy (Poecilia
reticulata)

Rainbow trout

(Oncorhynchus

mykiss)

0.71

1.24

24

0.63

0.00

0.68

0.6

198,443.9

69

4

PFDA, 7:3
FTCA, 8:2
FTUCA

Guppy (Poecilia
reticulata)

Fatliead minnow

(Pimephales

promelas)

0.85

0.35

27

0.78

0.00

0.39

1.2

140,225.3

66

4

PFBS

Indian bullfrog

(Euphlyctis

hexadactylus)

Bluegill (Lepomis
macrochirus)

0.88

0.02

4

0.86

0.01

0.29

37.9

113,000.0

67

5

PFBS

Indian bullfrog

(Euphlyctis

hexadactylus)

Rainbow trout

(Oncorhynchus

mykiss)

1.25

-0.12

5

0.78

0.01

0.59

28.3

19,926.0

57

5

8:2 FTUCA

Isopod (Asellus
aquaticus)

Daplinid (Daphnia
magna)

0.86

1.88

7

0.91

0.00

0.31

0.3

166,819.4

67

5

PFNA, PFHxA,
PFDA, PFBS,
PFBA, 7:3
FTCA, 8:2
FTUCA

Isopod (Asellus
aquaticus)

Bluegill (Lepomis
macrochirus)

1.09

-0.19

5

0.91

0.00

0.37

2.8

123,267.3

43

6

PFBS

Isopod (Asellus
aquaticus)

Mysid

(Americamysis
bahia)

0.79

1.69

4

0.89

0.00

0.31

0.0

6,982.6

83

4

PFBS

Isopod (Caecidotea
brevicauda)

Bluegill (Lepomis
macrochirus)

0.86

0.28

17

0.74

0.00

0.48

0.4

26,700.0

68

6

PFBS

E-9


-------
Predicted Species

Surrogate Species

Slope

Intercept

Degrees
of

Freedom
(N-2)

R2

p-value

Mean
Square
Error
(MSE)

Surrogate

model
minimum
value
(Hg/L)

Surrogate

model
maximum
value (ng/L)

Cross-
validation
Success
(%)

Taxonomic
Distance

PFAS
Compound

Isopod (Caecidotea
brevicauda)

Rainbow trout

(Oncorhynchus

mykiss)

0.78

0.41

19

0.70

0.00

0.52

0.6

82,000.0

57

6

PFDA, 7:3
FTCA, 8:2
FTUCA

Isopod (Caecidotea
brevicauda)

Fathead minnow

(Pimephales

promelas)

0.76

-0.05

8

0.73

0.00

0.49

1.2

20,922.0

60

6

PFBS

Lake trout

(Salvelinus

namaycush)

Bluegill (Lepomis
macrochirus)

0.69

0.80

20

0.72

0.00

0.27

2.8

180,000.0

77

4

PFBS

Lake trout

(Salvelinus

namaycush)

Rainbow trout

(Oncorhynchus

mykiss)

0.89

0.23

26

0.94

0.00

0.07

0.2

198,443.9

96

2

PFDA, 7:3
FTCA, 8:2
FTUCA

Lake trout

(Salvelinus

namaycush)

Fatliead minnow

(Pimephales

promelas)

0.77

0.04

12

0.66

0.00

0.34

44.4

133,000.0

79

4

PFBS

Largemouth bass

(Micropterus

sahnoides)

Bluegill (Lepomis
macrochirus)

1.03

-0.10

33

0.93

0.00

0.12

0.4

123,267.3

94

2

PFBS

Largemouth bass

(Micropterus

sahnoides)

Rainbow trout

(Oncorhynchus

mykiss)

0.94

0.04

32

0.89

0.00

0.21

0.0

95,857.7

91

4

PFDA, 7:3
FTCA, 8:2
FTUCA

Leech (Nephelopsis
obscura)

Daphnid (Daphnia
magna)

1.16

1.67

2

0.98

0.01

0.13

0.8

4,369.3

75

6

PFNA, PFDA,
7:3 FTCA

Mayfly (Drunella
grandis)

Rainbow trout

(Oncorhynchus

mykiss)

1.01

0.64

3

0.98

0.00

0.13

0.6

95,857.7

100

6

PFDA, 7:3
FTCA, 8:2
FTUCA

Medaka (Oryzias
latipes)

Rainbow trout

(Oncorhynchus

mykiss)

0.91

0.61

3

0.95

0.01

0.16

6.6

54,399.6

80

4

PFDA, 7:3
FTCA, 8:2
FTUCA

Medaka (Oryzias
latipes)

Fatliead minnow

(Pimephales

promelas)

0.86

0.60

6

0.98

0.00

0.05

2.7

1,420,000.0

88

4

PFBS

Midge (Chironomus
plumosus)

Mysid

(Americamysis
bahia)

0.74

0.93

13

0.78

0.00

0.66

0.0

87,600.0

67

5

PFBS

E-10


-------
Predicted Species

Surrogate Species

Slope

Intercept

Degrees
of

Freedom
(N-2)

R2

p-value

Mean
Square
Error
(MSE)

Surrogate

model
minimum
value
(Hg/L)

Surrogate

model
maximum
value (ng/L)

Cross-
validation
Success
(%)

Taxonomic
Distance

PFAS
Compound

Midge (Chironomus
tentans)

Daplinid (Daphnia
magna)

0.97

0.76

6

0.89

0.00

0.82

0.3

472,000.0

50

5

PFNA, PFDA,
7:3 FTCA

Midge (Chironomus
tentans)

Bluegill (Lepomis
macrochirus)

1.03

0.11

5

0.94

0.00

0.39

0.1

517,824.6

71

6

PFBS

Midge

(Paratanytarsus
dissiniilis)

Bluegill (Lepomis
macrochirus)

0.85

1.02

5

0.89

0.00

0.49

0.4

201,000.0

57

6

PFBS

Midge

(Paratanytarsus
dissiniilis)

Rainbow trout

(Oncorhynchus

mykiss)

0.89

1.03

8

0.90

0.00

0.33

0.6

1,330,000.0

80

6

PFDA, 7:3
FTCA, 8:2
FTUCA

Midge

(Paratanytarsus
dissiniilis)

Fatliead minnow

(Pimephales

promelas)

0.86

0.97

8

0.85

0.00

0.52

1.2

1,430,000.0

80

6

PFBS

Midge

(Paratanytarsus
dissiniilis)

Bullfrog

(Litliobates

catesbeianus)

1.09

-0.27

3

0.97

0.00

0.25

2.5

3,019,983.4

40

6

PFHxS, PFHxA

Midge

(Paratanytarsus
partlienogeneticus)

Daplinid (Daphnia
magna)

0.98

0.55

3

0.99

0.00

0.06

370.0

14,500,000.0

100

5

PFNA, PFHxA,
PFDA, PFBS,
PFBA, 7:3
FTCA, 8:2
FTUCA

Midge

(Paratanytarsus
partlienogeneticus)

Fatliead minnow

(Pimephales

promelas)

0.99

0.57

2

0.98

0.01

0.09

280.0

10,600,000.0

100

6

PFBS

Mississippi grass
shrimp

(Palaemonetes
kadiakensis)

Daplinid (Daphnia
magna)

0.76

0.47

20

0.63

0.00

0.72

0.3

58,000.0

64

5

PFNA, PFHxA,
PFDA, PFBS,
PFBA, 7:3
FTCA, 8:2
FTUCA

Mosquitofish
(Gambusia affmis)

Fatliead minnow

(Pimephales

promelas)

0.95

0.01

2

0.99

0.01

0.12

5.7

26,500,000.0

100

4

PFBS

Mosquitofish
(Gambusia affmis)

Bullfrog

(Litliobates

catesbeianus)

0.97

0.07

1

1.00

0.01

0.00

18600.0

20,900,000.0

N/A

5

PFHxS, PFHxA

E-ll


-------
Predicted Species

Surrogate Species

Slope

Intercept

Degrees
of

Freedom
(N-2)

R2

p-value

Mean
Square
Error
(MSE)

Surrogate

model
minimum
value
(Hg/L)

Surrogate

model
maximum
value (ng/L)

Cross-
validation
Success
(%)

Taxonomic
Distance

PFAS
Compound

Mozambique tilapia

(Oreochromis

mossambicus)

Fathead minnow

(Pimephales

promelas)

0.86

0.36

9

0.73

0.00

0.33

44.4

140,225.3

64

4

PFBS

Mrigal carp
(Cirrhinus mrigala)

Rainbow trout

(Oncorhynchus

mykiss)

0.76

1.03

4

0.71

0.04

0.43

0.6

1,624.7

67

4

PFDA, 7:3
FTCA, 8:2
FTUCA

Mrigal carp
(Cirrhinus mrigala)

Fathead minnow

(Pimephales

promelas)

1.11

-0.59

3

0.98

0.00

0.01

196.9

8,320.3

100

2

PFBS

Neosho mucket

(Lampsilis

rafmesqueana)

Daplinid (Daphnia
magna)

0.89

0.24

2

0.99

0.00

0.02

42.0

166,819.4

100

6

PFNA, PFHxA,
PFDA, PFBS,
PFBA, 7:3
FTCA, 8:2
FTUCA

Nile tilapia

(Oreochromis

niloticus)

Bluegill (Lepomis
macrochirus)

1.04

-0.04

1

1.00

0.00

0.00

2.3

6,969.1

N/A

3

PFBS

Northern leopard
frog (Lithobates
pipiens)

Bluegill (Lepomis
macrochirus)

0.80

1.13

1

1.00

0.02

0.01

7.9

180,000.0

N/A

5

PFBS

Northern leopard
frog (Lithobates
pipiens)

Rainbow trout

(Oncorhynchus

mykiss)

0.71

1.60

1

1.00

0.01

0.00

2.3

198,443.9

N/A

5

PFDA, 7:3
FTCA, 8:2
FTUCA

Northern pike (Esox
lucius)

Rainbow trout

(Oncorhynchus

mykiss)

1.04

-0.30

6

0.80

0.00

0.27

4.7

1,822.3

88

4

PFDA, 7:3
FTCA, 8:2
FTUCA

Oligochaete

(Limnodrilus

hoffmeisteri)

Zebrafish-embryo
(Danio rerio-
embryo)

1.17

-0.31

3

0.92

0.01

0.24

144.8

239,250.6

60

6

PFHxS, PFBA

Oligochaete

(Limnodrilus

hoffmeisteri)

Daplinid (Daplinia
magna)

0.71

1.77

10

0.81

0.00

0.36

3.2

281,641.4

67

6

PFNA, PFHxA,
PFDA, PFBS,
PFBA, 7:3
FTCA, 8:2
FTUCA

Oligochaete

(Limnodrilus

hoffmeisteri)

Rainbow trout

(Oncorhynchus

mykiss)

0.71

1.60

6

0.67

0.01

0.76

3.3

707,000.0

50

6

8:2 FTUCA

E-12


-------
Predicted Species

Surrogate Species

Slope

Intercept

Degrees
of

Freedom
(N-2)

R2

p-value

Mean
Square
Error
(MSE)

Surrogate

model
minimum
value
(Hg/L)

Surrogate

model
maximum
value (ng/L)

Cross-
validation
Success
(%)

Taxonomic
Distance

PFAS
Compound

Oligochaete

(Lumbriculus

variegatus)

Fathead minnow

(Pimephales

promelas)

0.94

0.28

14

0.82

0.00

0.43

1.2

140,225.3

75

6

PFBS

























PFNA, PFHxA,

Oligochaete (Tubifex
tubifex)

Daphnid (Daplinia
magna)

0.89

1.37

13

0.80

0.00

0.78

0.1

4,894,739.3

47

6

PFDA, PFBS,
PFBA, 7:3
FTCA, 8:2
FTUCA

Oligochaete (Tubifex
tubifex)

Bluegill (Lepomis
macrochirus)

0.72

0.67

7

0.72

0.00

0.84

0.1

113,000.0

56

6

PFBS

Oligochaete (Tubifex
tubifex)

Fathead minnow

(Pimephales

promelas)

1.04

-0.10

8

0.87

0.00

0.53

15.3

10,830,770.1

50

6

PFBS

Oriental river shrimp

(Macrobrachium

nipponense)

Daphnid (Daplinia
magna)

1.15

-0.84

3

0.99

0.00

0.06

11.0

281,641.4

100

5

PFNA, PFHxA,
PFDA, PFBS,
PFBA, 7:3
FTCA, 8:2
FTUCA

Paper pondshell

(Utterbackia

imbecillis)

Daphnid (Daplinia
magna)

0.89

0.28

11

0.98

0.00

0.08

14.1

8,694,505.9

100

6

PFNA, PFHxA,
PFDA, PFBS,
PFBA, 7:3
FTCA, 8:2
FTUCA

Paper pondshell

(Utterbackia

imbecillis)

Fatliead minnow

(Pimephales

promelas)

0.86

-0.16

9

0.65

0.00

0.85

134.2

10,830,770.1

18

6

PFBS

Peppered loach

(Lepidocephalichthys

guntea)

Fatliead minnow

(Pimephales

promelas)

0.78

0.72

1

1.00

0.03

0.00

196.9

43,785.6

N/A

3

PFBS

Pheasantshell
(Ortmanniana
pectorosa)

Daphnid (Daplinia
magna)

0.96

0.00

3

0.97

0.00

0.11

42.0

545,927.5

100

6

PFNA, PFHxA,
PFDA, PFBS,
PFBA, 7:3
FTCA, 8:2
FTUCA

Rainbow trout

(Oncorhynchus

mykiss)

Zebrafish (Danio
rerio)

1.31

-1.12

5

0.96

0.00

0.24

3.1

26,390.0

71

4

PFHxS, PFBA

E-13


-------
Predicted Species

Surrogate Species

Slope

Intercept

Degrees
of

Freedom
(N-2)

R2

p-value

Mean
Square
Error
(MSE)

Surrogate

model
minimum
value
(Hg/L)

Surrogate

model
maximum
value (ng/L)

Cross-
validation
Success
(%)

Taxonomic
Distance

PFAS
Compound

Rainbow trout

(Oncorhynchus

mykiss)

Zebrafish-embryo
(Danio rerio-
embryo)

1.16

-1.06

27

0.80

0.00

0.61

23.0

8,843,890.0

66

4

PFHxS, PFBA

Rainbow trout

(Oncorhynchus

mykiss)

Daplinid (Daphnia
magna)

0.71

1.03

377

0.63

0.00

0.84

0.1

14,500,000.0

60

6

PFNA, PFHxA,
PFDA, PFBS,
PFBA, 7:3
FTCA, 8:2
FTUCA

Rainbow trout

(Oncorhynchus

mykiss)

Bluegill (Lepomis
macrochirus)

0.95

0.03

314

0.89

0.00

0.22

0.1

7,100,000.0

91

4

PFBS

Rainbow trout

(Oncorhynchus

mykiss)

Mysid

(Americamysis
bahia)

0.66

1.35

201

0.62

0.00

0.72

0.0

113,000.0

68

6

PFBS

Rainbow trout

(Oncorhynchus

mykiss)

Fathead minnow

(Pimephales

promelas)

0.97

-0.20

146

0.87

0.00

0.29

0.2

26,500,000.0

87

4

PFBS

Rainbow trout

(Oncorhynchus

mykiss)

Bullfrog

(Lithobates

catesbeianus)

1.09

-0.71

9

0.95

0.00

0.38

2.5

20,900,000.0

73

5

PFHxS, PFHxA

Razorback sucker
(Xyrauchen texanus)

Bluegill (Lepomis
macrochirus)

0.81

0.01

6

0.67

0.01

0.48

53.9

150,750.0

75

4

PFBS

Razorback sucker
(Xyrauchen texanus)

Rainbow trout

(Oncorhynchus

mykiss)

0.78

0.91

6

0.87

0.00

0.19

3.3

43,073.0

88

4

PFDA, 7:3
FTCA, 8:2
FTUCA

Razorback sucker
(Xyrauchen texanus)

Fatliead minnow

(Pimephales

promelas)

0.98

-0.36

5

0.66

0.03

0.40

58.3

8,320.3

71

3

PFBS

Redear sunfish

(Lepomis

microlophus)

Bluegill (Lepomis
macrochirus)

1.03

-0.03

2

1.00

0.00

0.00

17.0

4,500.0

100

1

PFBS

Riceland prawn

(Macrobrachium

lanchesteri)

Rainbow trout

(Oncorhynchus

mykiss)

0.85

0.97

2

0.95

0.03

0.09

3.3

2,040.4

100

6

PFDA, 7:3
FTCA, 8:2
FTUCA

Rohu (Labeo rohita)

Bluegill (Lepomis
macrochirus)

0.94

0.77

4

0.94

0.00

0.21

0.8

7,326.2

83

4

PFBS

Rohu (Labeo rohita)

Rainbow trout

(Oncorhynchus

mykiss)

1.07

0.86

4

0.89

0.00

0.37

0.6

1,624.7

67

4

PFDA, 7:3
FTCA, 8:2
FTUCA

E-14


-------
















Surrogate

















Degrees





Mean

model

Surrogate

Cross-













of





Square

minimum

model

validation













Freedom





Error

value

maximum

Success

Taxonomic

PFAS

Predicted Species

Surrogate Species

Slope

Intercept

(N-2)

R2

p-value

(MSE)

(Hg/L)

value (ng/L)

(%)

Distance

Compound



Fathead minnow























Rohu (Labeo rohita)

(Pimephales
promelas)

0.98

0.32

4

0.87

0.01

0.30

2.3

8,320.3

83

2

PFBS

Shortiiose sturgeon

Rainbow trout





















PFDA, 7:3

(Acipenser

(Oncorhynchus

1.16

-0.58

3

0.98

0.00

0.06

28.3

95,857.7

100

5

FTCA, 8:2

brevirostriim)

mykiss)





















FTUCA

Silver perch
(Bidyanus bidyanus)

Rainbow trout

(Oncorhynchus

mykiss)

1.09

-0.03

1

1.00

0.01

0.00

1.4

7,075.2

N/A

4

PFDA, 7:3
FTCA, 8:2
FTUCA

Snipefly (Atherix
variegata)

Rainbow trout

(Oncorhynchus

mykiss)

0.94

0.88

1

1.00

0.02

0.00

0.6

68.4

N/A

6

PFDA, 7:3
FTCA

Sockeye salmon

Rainbow trout





















PFDA, 7:3

(Oncorhynchus

(Oncorhynchus

0.85

0.60

5

0.85

0.00

0.14

28.3

7,500.0

71

1

FTCA, 8:2

nerka)

mykiss)





















FTUCA

Southern leopard

Rainbow trout





















PFDA, 7:3

frog (Lithobates

(Oncorhynchus

1.28

-0.26

4

0.98

0.00

0.04

28.3

9,700.0

100

5

FTCA, 8:2

sphenocephalus)

mykiss)





















FTUCA

Spotfm chub

Rainbow trout





















PFDA, 7:3

(Erirnonax

(Oncorhynchus

1.20

-0.41

3

0.97

0.00

0.06

4.1

1,624.7

100

4

FTCA, 8:2

monachus)

mykiss)





















FTUCA

























PFNA, PFHxA,

Swamp lymnaea
(Lymnaea stagnalis)

Daphnid (Daphnia
magna)

0.94

0.20

6

0.96

0.00

0.23

14.1

8,694,505.9

88

6

PFDA, PFBS,
PFBA, 7:3
FTCA, 8:2
FTUCA

























PFNA, PFHxA,

Tadpole physa
(Phy sella gyrina)

Daphnid (Daphnia
magna)

0.96

0.11

8

0.96

0.00

0.20

14.1

8,694,505.9

80

6

PFDA, PFBS,
PFBA, 7:3
FTCA, 8:2
FTUCA

Tadpole physa
(Physella gyrina)

Fathead minnow

(Pimephales

promelas)

1.22

-1.60

6

0.84

0.00

0.58

813.9

10,830,770.1

25

6

PFBS

























PFNA, PFHxA,

Threeridge
(Amblema plicata)

Daphnid (Daphnia
magna)

0.95

-0.43

4

0.97

0.00

0.13

14.1

4,894,739.3

100

6

PFDA, PFBS,
PFBA, 7:3
FTCA, 8:2
FTUCA

E-15


-------
Predicted Species

Surrogate Species

Slope

Intercept

Degrees
of

Freedom
(N-2)

R2

p-value

Mean
Square
Error
(MSE)

Surrogate

model
minimum
value
(Hg/L)

Surrogate

model
maximum
value (ng/L)

Cross-
validation
Success
(%)

Taxonomic
Distance

PFAS
Compound

Threeridge
(Amblema plicata)

Fathead minnow

(Pimephales

promelas)

1.30

-2.81

3

0.98

0.00

0.13

813.9

10,830,770.1

100

6

PFBS

Vernal pool fairy
shrimp (Branchinecta
lynchi)

Daplinid (Daphnia
magna)

0.94

0.12

5

0.98

0.00

0.07

14.1

8,694,505.9

100

4

PFNA, PFHxA,
PFDA, PFBS,
PFBA, 7:3
FTCA, 8:2
FTUCA

Walleye (Sander
vitreus)

Bluegill (Lepomis
macrochirns)

0.74

0.54

7

0.78

0.00

0.22

4.3

6,722.1

67

3

PFBS

Walleye (Sander
vitreus)

Rainbow trout

(Oncorhynchus

mykiss)

0.68

0.60

7

0.69

0.01

0.28

4.7

16,235.6

56

4

PFDA, 7:3
FTCA, 8:2
FTUCA

Washboard

(Megalonaias

nervosa)

Daplinid (Daplinia
magna)

0.96

-0.23

9

0.97

0.00

0.13

14.1

8,694,505.9

91

6

PFNA, PFHxA,
PFDA, PFBS,
PFBA, 7:3
FTCA, 8:2
FTUCA

Water flea (Chydorns
sphaericus)

Daplinid (Daplinia
magna)

0.95

-0.20

7

0.98

0.00

0.06

8.9

977,589.9

100

3

PFNA, PFHxA,
PFDA, PFBS,
PFBA, 7:3
FTCA, 8:2
FTUCA

Water flea (Moina
macrocopa)

Daplinid (Daplinia
magna)

0.72

1.10

3

0.97

0.00

0.11

8.9

281,641.4

100

3

PFNA, PFHxA,
PFDA, PFBS,
PFBA, 7:3
FTCA, 8:2
FTUCA

Wavy rayed
lampmussel
(Lampsilis fasciola)

Daplinid (Daplinia
magna)

1.18

-1.28

1

0.99

0.05

0.01

166819.4

4,894,739.3

N/A

6

PFHxA

Western pearlshell

(Margaritifera

falcata)

Daplinid (Daplinia
magna)

0.88

0.27

8

0.96

0.00

0.16

14.1

8,694,505.9

80

6

PFNA, PFHxA,
PFDA, PFBS,
PFBA, 7:3
FTCA, 8:2
FTUCA

E-16


-------
Predicted Species

Surrogate Species

Slope

Intercept

Degrees
of

Freedom
(N-2)

R2

p-value

Mean
Square
Error
(MSE)

Surrogate

model
minimum
value
(Hg/L)

Surrogate

model
maximum
value (ng/L)

Cross-
validation
Success
(%)

Taxonomic
Distance

PFAS
Compound

Western toad
(Anaxyrus boreas)

Rainbow trout

(Oncorhynchus

mykiss)

1.33

-0.51

3

0.90

0.01

0.15

28.3

1,624.7

80

5

PFDA, 7:3
FTCA, 8:2
FTUCA

White sucker
(Catostomus
commersonii)

Bluegill (Lepomis
macrochirus)

1.05

-0.32

7

0.92

0.00

0.18

35.0

123,267.3

89

4

PFBS

White sucker
(Catostomus
commersonii)

Fathead minnow

(Pimephales

promelas)

1.15

-0.37

5

0.93

0.00

0.21

43.9

140,225.3

86

3

PFBS

Yellow perch (Perca
flavescens)

Bluegill (Lepomis
macrochirus)

0.98

-0.11

17

0.92

0.00

0.15

0.4

9,409.7

89

3

PFBS

Yellow perch (Perca
flavescens)

Rainbow trout

(Oncorhynchus

mykiss)

0.90

0.29

23

0.77

0.00

0.58

0.0

16,235.6

84

4

PFDA, 7:3
FTCA, 8:2
FTUCA

Yellow perch (Perca
flavescens)

Fatliead minnow

(Pimephales

promelas)

0.94

-0.38

12

0.74

0.00

0.54

1.2

20,922.0

64

4

PFBS

Zebrafish (Danio
rerio)

Daphnid (Daphnia
magna)

0.77

1.22

11

0.71

0.00

0.62

0.1

50,000.0

62

6

PFNA, PFHxA,
PFDA, PFBS,
PFBA, 7:3
FTCA, 8:2
FTUCA

Zebrafish (Danio
rerio)

Mysid

(Americamysis
bahia)

0.73

1.91

7

0.68

0.01

0.83

0.0

5,666.0

33

6

PFBS

Zebrafish (Danio
rerio)

Rainbow trout

(Oncorhynchus

mykiss)

0.73

0.96

5

0.96

0.00

0.14

0.2

229,100.0

100

4

PFDA, 7:3
FTCA, 8:2
FTUCA

Zebrafish (Danio
rerio)

Fatliead minnow

(Pimephales

promelas)

0.91

0.31

4

0.97

0.00

0.04

13.4

26,685.4

100

2

PFBS

Zebrafish-embryo
(Danio rerio-embryo)

Daphnid (Daphnia
magna)

0.66

1.72

43

0.67

0.00

0.87

0.1

46,500,000.0

56

6

PFNA, PFHxA,
PFDA, PFBS,
PFBA, 7:3
FTCA, 8:2
FTUCA

Zebrafish-embryo
(Danio rerio-embryo)

Rainbow trout

(Oncorhynchus

mykiss)

0.69

1.43

27

0.80

0.00

0.37

0.2

8,341,462.7

76

4

PFDA, 7:3
FTCA, 8:2
FTUCA

E-17


-------
Predicted Species

Surrogate Species

Slope

Intercept

Degrees
of

Freedom
(N-2)

R2

p-value

Mean
Square
Error
(MSE)

Surrogate

model
minimum
value
(Hg/L)

Surrogate

model
maximum
value (ng/L)

Cross-
validation
Success
(%)

Taxonomic
Distance

PFAS
Compound

Zebrafish-embryo
(Daiiio rerio-embryo)

Fathead minnow

(Pimephales

promelas)

0.93

0.38

41

0.93

0.00

0.20

4.0

70,200,000.0

81

2

PFBS

E-18


-------
Appendix F Derivation of PFAS Benchmarks using Scaled Data

As discussed in Section 3.0, PFAS acute values (typically reported as mg/L) can be
greater than those used to develop an ICE model (ICE database toxicity range lxlO"4 and 1x10s
(j,g/L) such that the input PFAS value of the surrogate would be outside the model domain. In
these situations, a user can either enter the measured toxicity value (LC50/EC50) into the ICE
model as [j,g/L and allow the regression to extrapolate beyond the range of the model or enter a
"scaled" toxicity value (i.e., enter the measured LC50 value as mg/L) (Raimondo et al. 2024).
Figure 3-1 provides an example data plots and calculation of an EC50 value for Daphnia magna
for PFBS for a value that falls beyond the model range, when calculated using an "extrapolation"
and "scaled" approach. Calculations presented in Section 5 of this document allowed
extrapolation beyond the range of the regression model, recognizing that such an extrapolation
has inherent uncertainties and can result in larger confidence intervals around the prediction than
would typically occur using the scaled approach. Within this section, we apply the alternative
scaled approach to explore calculating benchmark values for the eight PFAS using this
alternative method (Raimondo et al. 2024).

For the scaled approach, values are directly entered as [j,g/L in situations when the
surrogate species measured PFAS value falls within the range of the ICE model. In cases where
the surrogate species measured value is beyond the range of the ICE model, the value is then
entered as mg/L, so that it falls within the range of the model data. For example, if a hypothetical
concentration of 15 [j,g/L is beyond the range of the ICE model data for a chemical, it is then
entered as 15 mg/L to bring it within range of the model data. Values were also scaled in some
situations where the measured value falls within the model range but where the confidence
interval was large (>50 fold), and use of scaling resulted in acceptable confidence intervals.

F-l


-------
The use of scaled values has been validated through a comprehensive analysis of ICE
data, where it has been demonstrated that 3,104 values predicted in this manner had the same
level of accuracy as values entered into ICE models as |ig/L within and beyond the model
domain for models with slopes ranging from 0.66 - 1.33 (Raimondo et al. 2024). Raimondo et al.
(2024) also demonstrated that using ICE models in this manner does not violate any of the model
assumptions, which are: 1) they represent the relationship of inherent sensitivity between two
species, which is conserved across chemicals, mechanisms of action, and ranges of toxicity and
2) the nature of a contaminant that was tested on the surrogate reflects the nature of the
contaminant in the predicted species (e.g., effect concentration [ECso] or lethal concentration
[LCso]), percentage of active ingredient, technical grade, toxicity unit). Additionally, there are no
statistical assumptions of linear regressions (linear relationship, normality, homoscedasticity, no
auto-correlation, no or little multicollinearity; Dowdy et al. 2011) violated by this application.
The following sections present the calculations and resulting alternative benchmark values for
each of eight selected PFAS using the scaled approach. ICE models selected for use for the
derivation of recommended PFAS benchmarks are presented in Table E-l and the empirical data
used for this evaluation are presented in Appendix A.

F.l Derivation of Acute Water Benchmark for Perfluorobutanoic Acid (PFBA)

Quantitatively acceptable empirical data for PFBA were available for five species

comprising four genera and fulfilling three MDR groups (Table F-2). A total of 43 ICE models
were accepted for use. Of these, seven ICE models were not selected for use because empirical
data were available for the genera (e.g., models for Chydorus sphaericus and Daphnia magna).
Three other models were not used because there were acceptable models for more closely-related
surrogate species (e.g., Daphnia magna predicting for Pimephalespromelas). One additional
model (Danio rerio predicting for Pimephales promelas) was not used because the measured

F-2


-------
EC50 values fell outside of the ICE model range and other acceptable models for taxonomically-
related species with measured EC50 values within the ICE model range were used. A total of 32
ICE models were selected for use, resulting in ICE models predicting 31 SMAVs, representing
29 genera (Tables F-l and F-2).

The combined empirical and ICE data resulted in 33 GMAVs that collectively fulfill the
eight MDR groups for deriving a freshwater criterion as defined by the Aquatic Life Criteria
Guidelines (U.S. EPA 1985). The ranked GMAVs for these combined data along with the MDR
met by each GMAV are summarized in Table F-2. GMAVs for the four most sensitive genera
were within a factor of 13.9 of each other (Table F-3). The freshwater FAV (the 5th percentile of
the genus sensitivity distribution) for PFBA is 347.1 mg/L (Table F-3), which is lower than all
of the GMAVs except for one tested species (the rotifer, Brachionus calyciflorus, GMAV=110
mg/L). The FAV was divided by two to obtain a preliminary value of 173.5 mg/L PFBA and
then adjusted by the carboxylic acid application factor (15.8) to obtain the alternative freshwater
acute water column benchmark magnitude of 11 mg/L PFBA (rounded to two significant
figures). This value is expected to be protective of 95% of freshwater genera exposed to PFBA
under short-term conditions of one-hour duration, if the one-hour average magnitude is not
exceeded more than once in three years (Figure F-l).

F-3


-------
Table F-l. Acceptable models for ICE-estimated Species Sensitivity to PFBA using the scaling approach for data outside of the
model bounds as in cases indicated.

Bold predicted EC50S used for SMAY calculations.			

Predicted Species

Surrogate

ICE Model Value Range
(mg/L)

Predicted EC50
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Entered
ECso
(units)

Minimum

Maximum

Amphipod (Crangonyx
pseudogracilis)

Daphnid (Daphnia
magna)

5,251**

5,251
(mg/L)

0.0008

166.8

44,908.2

(10,018-201,313)

Amphipod (Gammarus
fasciatus)

Daphnid (Daphnia
magna)

5,251**

5,251
(mg/L)

0.0002

5,000

3,060.4

(1,474.4-6,352.5)

Amphipod (Gammarus
pseudolimnaeus)

Daphnid (Daphnia
magna)

5,251**

5,251
(mg/L)

0.0001

68.3

4,490.4

(1,523.0-13,240)

Beaver-tail fairy shrimp
(Thamnocephalus platyurus)

Daphnid (Daphnia
magna)

5,251

5,251,000
(Hg/L)

0.0003

8,694.5

2,248.7

(1,108-4,563)

Bluegill (Lepomis
macrochirus)

Daphnid (Daphnia
magna)

5,251

5,251,000
(Ug/L)

0.0001

46,278.3

959.02

(507.98-1,810.6)

Channel catfish (Ictalurus
punctatus)

Zebrafish-embryo (Danio
rerio-embryo)

13,779**

13,779
(mg/L)

0.145

304.5

13,361.7

(6,856.1-26,037)

Common carp (Cyprinus
carpio)

Zebrafish-embryo (Danio
rerio-embryo)

13,779**

13,779
(mg/L)

0.086

34.19

7,908.4

(1,225.4-51,039)

Daphnid (Ceriodaphnia dubia)

Daphnid (Daphnia
magna)

5,251

5,251,000
(Hg/L)

0.0003

46,278.3

4,100.3

(2,125.4-7,909.9)

Daphnid (Daphnia
longispina)*

Daphnid (Daphnia
magna)

5,251**

5,251
(mg/L)

0.009

10.36

8,995.3
(1,641.2-49,301)

Daphnid (Daphnia magna)*

Zebrafish (Danio rerio)

13,779**

13,779
(mg/L)

0.003

36.91

3,291.4
(831.19-13,034)

Daphnid (Daphnia pulex)*

Daphnid (Daphnia
magna)

5,251**

5,251
(mg/L)

0.0002

4,894.7

3,957.7
(2,788.6-5,616.9)

Daphnid (Daphnia pulicaria)*

Daphnid (Daphnia
magna)

5,251**

5,251
(mg/L)

0.014

281.6

4,361.5
(1,223.6-15,546)

Daphnid (Simocephalus
serrulatus)

Daphnid (Daphnia
magna)

5,251**

5,251
(mg/L)

0.0002

7.2

2,939.0

(964.77-8,953.2)

F-4


-------
Predicted Species

Surrogate

ICE Model Value Range
(mg/L)

Predicted EC50
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Entered
ECso
(units)

Minimum

Maximum

Fathead minnow (Pimephales
promelas)

Daphnid (Daphnia
magna)

5,251

5,251,000
(Hg/L)

0.0002

46,500

l,174.4b
(655.01-2,105.8)

Fathead minnow (Pimephales
promelas)

Zebrafish (Danio rerio)

13,779**

13,779
(mg/L)

0.018

25.08

15,343.0C
(7,383.2-31,884)

Fathead minnow (Pimephales
promelas)

Zebrafish-embryo (Danio
rerio-embryo)

13,779

13,779,000
(Hg/L)

0.023

54,579

11,532.0

(6,189.6-21,485)

Fatmucket (Lampsilis
siliquoidea)

Daphnid (Daphnia
magna)

5,251

5,251,000
(Ug/L)

0.014

8,694.5

2,013.5

(809.37-5,008.9)

Flagfish (Jordanella floridae)

Zebrafish (Danio rerio)

13,779**

13,779
(mg/L)

7.797

649.3

18,640.2

(10,316-33,681)

Goldfish (Carassius auratus)

Zebrafish-embryo (Danio
rerio-embryo)

13,779**

13,779
(mg/L)

0.145

304.5

23,117.1

(14,833-36,028)

Green floater (Lasmigona
subviridis)

Daphnid (Daphnia
magna)

5,251**

5,251
(mg/L)

0.014

166.8

2,645.9

(481.11-14,552)

Isopod (Asellus aquaticus)

Daphnid (Daphnia
magna)

5,251**

5,251
(mg/L)

0.0003

166.8

116,979.2

(32,795-417,264)

Midge (Paratanytarsus
parthenogeneticus)

Daphnid (Daphnia
magna)

5,251

5,251,000
(Hg/L)

0.37

14,500

12,921.0

(3,394.0-49,189)

Mississippi grass shrimp
(Palaemonetes kadiakensis)

Daphnid (Daphnia
magna)

5,251**

5,251
(mg/L)

0.0003

58

1,932.2

(372.75-10,016)

Neosho mucket (Lampsilis
rafinesqueana)

Daphnid (Daphnia
magna)

5,251**

5,251
(mg/L)

0.042

166.8

3,613.7

(1,572.1-8,306.4)

Oligochaete (Limnodrilus
hoffmeisteri)

Daphnid (Daphnia
magna)

5,251**

5,251
(mg/L)

0.003

281.6

25,412.0

(9,374.1-68,889)

Oligochaete (Limnodrilus
hoffmeisteri)

Zebrafish-embryo (Danio
rerio-embryo)

13,779**

13,779
(mg/L)

0.145

239.3

35,536.4b
(6,336.9-199,284)

Oligochaete (Tubifex tubifex)

Daphnid (Daphnia
magna)

5,251**

5,251
(mg/L)

0.0001

4,894.7

49,159.1

(10,555-228,948)

Oriental river shrimp
(Macrobrachium nipponense)

Daphnid (Daphnia
magna)

5,251**

5,251
(mg/L)

0.011

281.6

2,680.1

(1,224.4-5,866.5)

F-5


-------
Predicted Species

Surrogate

ICE Model Value Range
(mg/L)

Predicted EC50
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Entered
ECso
(units)

Minimum

Maximum

Paper pondshell (Utterbackia
imbecillis)

Daphnid (Daphnia
magna)

5,251

5,251,000
(Hg/L)

0.014

8,694.5

1,748.6

(862.33-3,545.6)

Pheasantshell (Ortmanniana
pectorosa)

Daphnid (Daphnia
magna)

5,251**

5,251
(mg/L)

0.042

545.9

3,775.7

(1,269.6-11,229)

Rainbow trout (Oncorhynchus
mykiss)

Daphnid (Daphnia
magna)

5,251

5,251,000
(Hg/L)

0.0001

14,500

677.8b
(396.18-1,159.4)

Rainbow trout (Oncorhynchus
mvkiss)

Zebrafish (Danio rerio)

13,779**

13,779
(mg/L)

0.003

26.39

20,148.4

(5,451.2-74,471)

Rainbow trout (Oncorhynchus
mykiss)

Zebrafish-embryo (Danio
rerio-embryo)

13,779**

13,779
(mg/L)

0.023

8,843.9

5,447.6

(2,518.2- 11,785)

Swamp lymnaea (Lymnaea
stagnalis)

Daphnid (Daphnia
magna)

5,251

5,251,000
(Ug/L)

0.014

8,694.5

3,490.7

(670.81-18,165)

Tadpole physa (Physella
gyrina)

Daphnid (Daphnia
magna)

5,251

5,251,000
(Hg/L)

0.014

8,694.5

3,740.1

(1,061.7-13,176)

Threeridge (Amblema plicata)

Daphnid (Daphnia
magna)

5,251**

5,251
(mg/L)

0.014

4,894.7

1,262.4

(480.17-3,318.7)

Vernal pool fairy shrimp
(Branchinecta lynchi)

Daphnid (Daphnia
magna)

5,251

5,251,000
(Hg/L)

0.014

8,694.5

2,852.6

(949.6-8,569.8)

Washboard (Megalonaias
nervosa)

Daphnid (Daphnia
magna)

5,251

5,251,000
(Ug/L)

0.014

8,694.5

1,759.2

(742.37-4,168.8)

Water flea (Chydorus
sphaericus)*

Daphnid (Daphnia
magna)

5,251**

5,251
(mg/L)

0.009

977.6

2,151.3
(1,364.7-3391.2)

Water flea (Moina
macrocopa)

Daphnid (Daphnia
magna)

5,251**

5,251
(mg/L)

0.009

281.6

5,989.5

(1,810.5-19,815)

Western pearlshell
(Margaritifera falcata)

Daphnid (Daphnia
magna)

5,251

5,251,000
(Hg/L)

0.014

8,694.5

1,528.3

(497.59-4,694.2)

Zebrafish (Danio rerio)*

Daphnid (Daphnia
magna)

5,251**

5,251
(mg/L)

0.0001

50

12,209.2
(3,373.7-44,185)

Zebrafish-embryo (Danio
rerio-embryo)*

Daphnid (Daphnia
magna)

5,251

5,251,000
(Hg/L)

0.0001

46,500

1,391.4
(413.39-4,683.2)

* Acceptable models that were not used because genus level empirical data were available.

F-6


-------
**Measured EC50 falls outside range of ICE model and a "scaled" EC50 was therefore entered in ICE model.
a Saltwater surrogate species were not used.

b Not used because other more closely taxonomically-related models were available.

0 Not used because other more closely taxonomically-related models where the measured surrogate EC50 values fell within the ICE model range were available.

F-7


-------
Table F-2. Ranked PFBA Genus Mean Acute Values.

Values in bold and highlighted are derived from empirical PFBA toxicity tests with the species.

Rank

MDR

Group1

Name

Species

SMAV

GMAV

Percentile
Rank

1

H

Rotifer

Brachionus calyciflorus

110

110

0.03

2

B

Bluegill

Lepomis macrochirus

959

959

0.06

3

G

Threeridge

Amblema plicata

1,262**

1,262

0.09

4

G

Western pearlshell

Margaritifera falcata

1,528

1,528

0.12

5

G

Paper pondshell

Utterbackia imbecillis

1,749

1,749

0.15

6

G

Washboard

Megalonaias nen'osa

1,759

1,759

0.18

7

E

Mississippi grass shrimp

Palaemonetes kadiakensis

1,932**

1,932

0.21

8

E

Beaver-tail fairy shrimp

Thamnocephalus platvurus

2,249

2,249

0.24

9

G

Green floater

Lasmigona subviridis

2,646**

2,646

0.26

10

E

Oriental river shrimp

Macrobrachium
nipponense

2,680**

2,680

0.29

11

G

Neosho mucket

Lampsilis rafinesqueana

3,614**

2,697

0.32

G

Fatmucket

Lampsilis siliquoidea

2,013

12

E

Vernal pool fairy shrimp

Brcmchinecta Ivnchi

2,853

2,853

0.35

13

D

Daphnid

Simocephalus serrulatus

2,939**

2,939

0.38

14

G

Swamp lymnaea

Lymnaea stagnalis

3,491

3,491

0.41

15

E

Amphipod

Gammarus fasciatus

3,060**

3,707

0.44

E

Amphipod

Gammarus pseudolimnaeus

4,490**

16

G

Tadpole physa

Phvsella gvrina

3,740

3,740

0.47

17

G

Pheasantshell

Ortmanniana pectorosa

3,776**

3,776

0.50

18

D

Daphnid

Ceriodaphnia dubia

4,100

4,100

0.53

19

D

Daphnid

Chydorus sphaericus

>4,280.8

>4,280.8

0.56

20

D

Daphnid

Daphnia magna

4,741

4,741

0.59

D

Daphnid

Daphnia pulicaria

>l,006a

21

D

Water flea

Moina macrocopa

5,989**

5,989

0.62

22

B

Common carp

Cyprinus carpio

7,908**

7,908

0.65

23

A

Rainbow trout

Oncorhvnchus mvkiss

10,477**

10,477

0.68

24

B

Fathead minnow

Pimephales promelas

11,532

11,532

0.71

25

F

Midge

Paratanytarsus
parthenogeneticus

12,921

12,921

0.74

26

C

Channel catfish

Ictalurus punctatus

13,361**

13,361

0.76

27

B

Zebrafish

Danio rerio

13,779

13,779

0.79

28

C

Flagfish

Jordanella floridae

18,640**

18,640

0.82

29

B

Goldfish

Carassius auratus

23,117**

23,117

0.85

30

H

Oligochaete

Linmodrilus hoffmeisteri

25,412**

25,412

0.88

31

E

Amphipod

Crangonyx pseudogracilis

44,908**

44,908

0.91

32

H

Oligochaete

Tubifex tubifex

49,159**

49,159

0.94

33

E

Isopod

Asellus aquaticus

116,979**

116,979

0.97

**Measured EC50 falls outside range of ICE model and a "scaled" EC50 was therefore entered in ICE model,
a Not used in GMAV calculation, because value represents a greater than low value
1: Freshwater MDR Groups

A)	The family Salmonidae in the class Osteichthyes

B)	A second family in the class Osteichthyes, preferably a commercially or recreationally important

warmwater species (e.g., bluegill, channel catfish, etc.)

C)	A third family in the phylum Chordata (may be in the class Osteichthyes or may be an amphibian, etc.)

F-8


-------
D)	A planktonic crustacean (e.g., cladoceran, copepod, etc.)

E)	Abentliic crustacean (e.g., ostracod, isopod, amphipod, crayfish, etc.)

F)	An insect (e.g., mayfly, dragonfly, damselfly, stonefly, caddisfly, mosquito, midge, etc.)

G)	A family in a phylum other than Arthropoda or Chordata (e.g., Rotifera, Annelida, Mollusca, etc.)

H)	A family in any order of insect or any phylum not already represented.

Table F-3. PFBA Protective Aquatic Life Acute Benchmark.

Bold values represent genera for which empirical toxicity data were available.

Calculated Freshwater FAY based on 4 lowest values; N=33 GMAVs total





Benchmark calculated by dividing the FAY by 2 and by the carboxylic acid application factor (15.8)





GMAV









Rank

Genus

(mg/L)

ln(GMAV)

ln(GMAV)2

P=R/(N+1)

sqrt(P)

1

Brachionus

110.0

4.70

22.09

0.029

0.171

2

Lepomis

959.0

6.87

47.14

0.059

0.243

3

Amblema

1262.4

7.14

50.99

0.088

0.297

4

Margaritifera

1528.3

7.33

53.76

0.118

0.343



£ (Sum):

26.04

173.98

0.29

1.05







P = cumulative probability











R = rank













N = number of GMAVs





S2 =

273.69



S = slope







L =

2.150



L = X-axis intercept





A =

5.849



A = InFAV







FAV =

347.1











FAV/2 =

173.5 mg/L (Preliminary Value)







Adjustment =

173.5 / 15.8 = 10.98 (Preliminary Value / Carboxylic Acid Application Factor)

Benchmark =

11 mg/L PFBA (rounded to two significant figures)





F-9


-------
1.00 -r

0.90

.2 0.80 4

*2 0.70 -
>

0.60
0.50

5

«

06

J 0.40 4
| 0.30 4

OI

Ph

0.20 4
0.10
0.00

1

o
+
~

A

O

Fish (Empirical)

Fish (WeblCE)

Insect (WeblCE)
Invertebrate (Empirical)
Invertebrate (WeblCE)
Mollusk (WeblCE)
Preliminary Value
¦Acute Benchmark

A Asellus
A Tubifex
A Crangonyx
A Limnodrilus
O Carassius
o Jordanella
9 Danio
O Ictalurus
+ Paratanytarsus
O Pimephales
O Oncorhynchus
O Cyprinus
A Moina
A Daphnia
A Chydorus
A Ceriodaphnia
O Ortmaimiana
O Physella
A Gammarus
O Lymnaea
A Simocephalus
A Branchinecta
O Lampsilis
A Macrobrachium
O Lasmigona
A Thamnocephalus
A Palaemonetes
O Megalonaias
O Utterbackia
O Margaritifera
O Amblema
O Lepomis

10	100	1,000	10,000

Genus Mean Acute Value (mg/L PFBA)

100,000

1,000,000

Figure F-l. Ranked Acute PFBA GMAVs Used for the Aquatic Life Acute Benchmark
Calculation.

F.2 Derivation of Acute Water Benchmark for Perfluorohexanoic Acid (PFHxA)

Quantitatively acceptable empirical data for PFHxA were available for four species

comprising three genera and fulfilling three MDR groups (Table F-5). A total of 43 ICE models
were accepted for use. Of these, four ICE models were not selected for use because empirical
data were available for the genus (e.g., models for Daphnia longispina and Daphnia magna).
Three other models were not used because there were acceptable models for more closely-related
surrogate species (e.g., Daphnia magna predicting for Lepomis macrochirus). One additional
model (Daphnia magna predicting for Danio rerio) was not used because the measured EC so
values fell outside of the ICE model range and other acceptable models for taxonomically-related

F-10


-------
species with measured EC50 values within the ICE model range were used. A total of 35 ICE
models were selected for use, resulting in ICE models predicting 35 SMAVs representing 30
genera (Tables F-4 and F-5).

The combined empirical and ICE data resulted in 33 GMAVs that collectively fulfill the
eight MDR groups for deriving a freshwater criterion as defined by the Aquatic Life Criteria
Guidelines (U.S. EPA 1985). The ranked GMAVs for these combined data along with the MDR
met by each GMAV are summarized in Table F-5. GMAVs for the four most sensitive genera
were within a factor of 2.7 of each other (Table F-6). The freshwater FAV (the 5th percentile of
the genus sensitivity distribution) for PFHxA is 189.1 mg/L (Table F-6), which is lower than all
of the GMAVs except for one tested species (the rotifer, Brachionus calyciflorus, GMAV=140
mg/L). The FAV was divided by two to obtain a preliminary value of 94.57 mg/L PFHxA and
then adjusted by the carboxylic acid application factor (15.8) to obtain the freshwater acute water
column benchmark magnitude of 6.0 mg/L PFHxA (rounded to two significant figures). This
value is expected to be protective of 95% of freshwater genera exposed to PFHxA under short-
term conditions of one-hour duration, if the one-hour average magnitude is not exceeded more
than once in three years (Figure F-2).

F-ll


-------
Table F-4. Acceptable models for ICE-estimated Species Sensitivity to PFHxA using the scaling approach for data outside of
the model bounds as in cases indicated.

Bold predicted EC50S used for SMAY calculations.			

Predicted Species

Surrogate

ICE Model Value Range
(mg/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Entered
ECso
(units)

Minimum

Maximum

Amphipod (Crangonyx
pseudogracilis)

Daphnid (Daphnia magna)

1,048**

1,048
(mg/L)

0.0008

166.8

10,313.0

(3,000.6-35,445)

Amphipod (Gammarus
fasciatus)

Daphnid (Daphnia magna)

1,048

1,048,000
(Hg/L)

0.0002

5,000

234.7

(64.5-853.7)

Amphipod (Gammarus
pseudolimnaeus)

Daphnid (Daphnia magna)

1,048**

1,048
(mg/L)

0.0001

68.3

1,026.8

(431.3-2,444.8)

Beaver-tail fairy shrimp
(Thamnocephalus platyurus)

Daphnid (Daphnia magna)

1,048

1,048,000
(Hg/L)

0.0003

8,694.5

517.3

(280.8-953.1)

Bluegill (Lepomis
macrochirus)

Bullfrog (Lithobates
catesbeianus)

1,105**

1,105
(mg/L)

0.003

233

766.4

(378-1,553.6)

Bluegill (Lepomis
macrochirus)

Daphnid (Daphnia magna)

1,048

1,048,000
(Hg/L)

0.0001

46,278

303.5b
(176-523.2)

Channel catfish (Ictalurus
punctatus)

Bullfrog (Lithobates
catesbeianus)

1,105

1,105,000
(Ug/L)

0.003

20,900

719.8

(362.4-1,429.7)

Daphnid (Ceriodaphnia
dubia)

Daphnid (Daphnia magna)

1,048

1,048,000
(Hg/L)

0.0003

46,278

785.2

(446.2-1,381.6)

Daphnid (Daphnia
longispina)*

Daphnid (Daphnia magna)

1,048**

1,048
(mg/L)

0.009

10.36

1,267.8
(334.3-4,808.1)

Daphnid (Daphnia magna)*

Bullfrog (Lithobates
catesbeianus)

1,105

1,105,000
(Hg/L)

0.003

20,900

424.1
(68.5-2,627.9)

Daphnid (Daphnia pulex)*

Daphnid (Daphnia magna)

1,048

1,048,000
(Hg/L)

0.0002

4,894.7

841.4
(444.8-1,591.6)

Daphnid (Daphnia pulicaria)*

Daphnid (Daphnia magna)

1,048**

1,048
(mg/L)

0.014

281.6

783.4
(181.5-3,381.5)

Daphnid (Simocephalus
serrulatus)

Daphnid (Daphnia magna)

1,048**

1,048
(mg/L)

0.0002

7.2

648.9

(269.3-1,563.9)

F-12


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Predicted Species

Surrogate

ICE Model Value Range
(mg/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Entered
ECso
(units)

Minimum

Maximum

Daphnid (Simocephalus
vetulus)

Daphnid (Daphnia magna)

1,048**

1,048
(mg/L)

0.0001

166.8

753.7

(136-4,175.6)

Fathead minnow (Pimephales
promelas)

Bullfrog (Lithobates
catesbeianus)

1,105

1,105,000
(Ug/L)

0.003

20,900

1,083.6

(468.5-2,506.4)

Fathead minnow (Pimephales
promelas)

Daphnid (Daphnia magna)

1,048

1,048,000
(Hg/L)

0.0002

46,500

384.7b
(235.8-627.7)

Fatmucket (Lampsilis
siliquoidea)

Daphnid (Daphnia magna)

1,048

1,048,000
(Ug/L)

0.014

8,694.5

470.8

(220.2-1,006.8)

Goldfish (Carassius auratus)

Bullfrog (Lithobates
catesbeianus)

1,105

1,105,000
(Hg/L)

0.003

20,900

1,027.7

(405.9-2,602.1)

Green floater (Lasmigona
subviridis)

Daphnid (Daphnia magna)

1,048**

1,048
(mg/L)

0.014

166.8

901.5

(216-3,761.6)

Isopod (Asellus aquaticus)

Daphnid (Daphnia magna)

1,048**

1,048
(mg/L)

0.0003

166.8

29,446.1

(9,967-86,994)

Midge (Paratanytarsus
dissimilis)

Bullfrog (Lithobates
catesbeianus)

1,105A

1,105
(mg/L)

0.003

3,020.0

1,069.7

(176.6-6,480.9)

Midge (Paratanytarsus
parthenogeneticus)

Daphnid (Daphnia magna)

1,048

1,048,000
(Hg/L)

0.37

14,500

2,676.6

(917.7-7,806.3)

Mississippi grass shrimp
(Palaemonetes kadiakensis)

Daphnid (Daphnia magna)

1,048**

1,048
(mg/L)

0.0003

58

569.4

(155.9-2,080.1)

Mosquitofish (Gambusia
affinis)

Bullfrog (Lithobates
catesbeianus)

1,105

1,105,000
(Hg/L)

18.6

20,900

805.1

(394.5-1,643.4)

Neosho mucket (Lampsilis
rafinesqueana)

Daphnid (Daphnia magna)

1,048**

1,048
(mg/L)

0.042

166.8

860.1

(421.3-1,755.9)

Oligochaete (Limnodrilus
hoffineisteri)

Daphnid (Daphnia magna)

1,048**

1,048
(mg/L)

0.003

281.6

8,114.2

(3,337.6-19,727)

Oligochaete (Tubifex tubifex)

Daphnid (Daphnia magna)

1,048A

1,048
(mg/L)

0.0001

4,894.7

11,637.0

(3,223.3-42,013)

Oriental river shrimp
(Macrobrachium nipponense)

Daphnid (Daphnia magna)

1,048**

1,048
(mg/L)

0.011

281.6

421.3

(176.7-1004.7)

F-13


-------
Predicted Species

Surrogate

ICE Model Value Range
(mg/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Entered
ECso
(units)

Minimum

Maximum

Paper pondshell (Utterbackia
imbecillis)

Daphnid (Daphnia magna)

1,048

1,048,000
(Hg/L)

0.014

8,694.5

418.5

(231.7-755.9)

Pheasantshell (Ortmanniana
pectorosa)

Daphnid (Daphnia magna)

1,048**

1,048
(mg/L)

0.042

545.9

803.4

(252.6-2,555.1)

Rainbow trout
(Oncorhynchus mykiss)

Bullfrog (Lithobates
catesbeianus)

1,105

1,105,000
(Hg/L)

0.003

20,900

713.5

(200.3-2,541.4)

Rainbow trout
(Oncorhynchus mykiss)

Daphnid (Daphnia magna)

1,048

1,048,000
(ug/L)

0.0001

14,500

214.2b
(135.8-337.8)

Swamp lymnaea (Lymnaea
stagnalis)

Daphnid (Daphnia magna)

1,048

1,048,000
(Hg/L)

0.014

8,694.5

762.0

(188.8-3,075.8)

Tadpole physa (Physella
gyrina)

Daphnid (Daphnia magna)

1,048

1,048,000
(ug/L)

0.014

8,694.5

794.5

(277.3-2,276.3)

Threeridge (Amblema
plicata)

Daphnid (Daphnia magna)

1,048

1,048,000
(Hg/L)

0.014

4,894.7

191.3

(48-761.8)

Vernal pool fairy shrimp
(Branchinecta lynchi)

Daphnid (Daphnia magna)

1,048

1,048,000
(ug/L)

0.014

8,694.5

623.8

(247-1,575.4)

Washboard (Megalonaias
nervosa)

Daphnid (Daphnia magna)

1,048

1,048,000
(Hg/L)

0.014

8,694.5

372.3

(181.1-765.2)

Water flea (Chydorus
sphaericus)

Daphnid (Daphnia magna)

1,048**

1,048
(mg/L)

0.009

977.6

465.4

(271.8-796.9)

Water flea (Moina
macrocopa)

Daphnid (Daphnia magna)

1,048**

1,048
(mg/L)

0.009

281.6

1,876.3

(633.2-5,560.2)

Wavyrayed lampmussel
(Lampsilis fasciola)

Daphnid (Daphnia magna)

1,048

1,048,000
(Ug/L)

166.8

4,894.7

686.2

(127.8-3,684.7)

Western pearlshell
(Margaritifera falcata)

Daphnid (Daphnia magna)

1,048

1,048,000
(Hg/L)

0.014

8,694.5

370.2

(145.8-940.1)

Zebrafish (Danio rerio)

Daphnid (Daphnia magna)

1,048**

1,048
(mg/L)

0.0001

50

3,520.9C
(1,155.2-10,731)

Zebrafish-embryo (Danio
rerio-embryo)

Daphnid (Daphnia magna)

1,048

1,048,000
(Hg/L)

0.0001

46,500

481.7

(172.4-1,346.1)

* Acceptable models that were not used because genus level empirical data were available.

F-14


-------
**Measured EC50 falls outside range of ICE model and a "scaled" EC50 was therefore entered in ICE model.

A |ig/L CI greater than 50-fold. Used mg/L with acceptable CI.
a Saltwater surrogate species were not used.

b Not used because other more closely taxonomically-related models were available.

0 Not used because other more closely taxonomically-related models where the measured surrogate EC50 values fell within the ICE model range were available.

F-15


-------
Table F-5. Ranked PFHxA Genus Mean Acute Values.

Values in bold and highlighted are derived from empirical PFHxA toxicity tests with the species.

Rank

MDR

Group1

Name

Species

SMAV

GMAV

Percentile
Rank

1

H

Rotifer

Brachionus calyciflorus

140.0

140.0

0.03

2

G

Threeridge

Amblema plicata

191.3

191.3

0.06

3

G

Western pearlshell

Margaritifera falcata

370.2

370.2

0.09

4

G

Washboard

Megalonaias nen'osa

372.3

372.3

0.12

5

G

Paper pondshell

Utterbackia imbecillis

418.5

418.5

0.15

6

E

Oriental river shrimp

Macrobrachium
nipponense

421.3**

421.3

0.18

7

D

Water flea

Chydorus sphaericus

465.4**

465.4

0.21

8

B

Zebrafish

Danio rerio

481.7

481.7

0.24

9

E

Amphipod

Gammarus fasciatus

234.7

490.9

0.26

E

Amphipod

Gammarus pseudolimnaeus

1,027**

10

E

Beaver-tail fairy shrimp

Thamnocephalus platvurus

517.3

517.3

0.29

11

E

Mississippi grass shrimp

Palaemonetes kadiakensis

569.4**

569.4

0.32

12

E

Vernal pool fairy shrimp

Brcmchinecta Ivnchi

623.8

623.8

0.35

13

G

Wavyrayed lampmussel

Lampsilis fasciola

686.2

652.6

0.38

G

Neosho mucket

Lampsilis rafinesqueana

860.1**

G

Fatmucket

Lampsilis siliquoidea

470.8

14

D

Daphnid

Simocephalus serrulatus

648.9**

699.3

0.41

D

Daphnid

Simocephalus vetulus

753.7**

15

A

Rainbow trout

Oncorhvnchus mvkiss

713.5

713.5

0.44

16

B

Channel catfish

Ictalurus punctatus

719.8

719.8

0.47

17

G

Swamp lymnaea

Lymnaea stagnalis

762.0

762.0

0.50

18

B

Bluegill

Lepomis macrochirus

766.4**

766.4

0.53

19

D

Daphnid

Ceriodaphnia dubia

785.2

785.2

0.56

20

G

Tadpole physa

Phvsella gyrina

794.5

794.5

0.59

21

G

Pheasantshell

Ortmanniana pectorosa

803.4**

803.4

0.62

22

B

Mosquitofish

Gambusia affmis

805.1

805.1

0.65

23

G

Green floater

Lasmigona subviridis

901.5**

901.5

0.68

24

C

Bullfrog

Lithobates catesbeianus

1,105

915.2

0.71

C

Green frog

Lithobates clamitans

758.0

25

B

Goldfish

Carassius auratus

1,028

1,028

0.74

26

D

Daphnia

Daphnia magna

1,048

1,048

0.76

27

B

Fathead minnow

Pimephales promelas

1,084

1,084

0.79

28

F

Midge

Paratanvtarsus dissimilis

1,070

1,692

0.82

F

Midge

Paratanytarsus
parthenogeneticus

2,677

29

D

Water flea

Moina macrocopa

1,876**

1,876

0.85

30

H

Oligochaete

Limnodrilus hoffmeisteri

8,114**

8,114

0.88

31

E

Amphipod

Crangonyx pseudogracilis

10,313**

10,313

0.91

32

H

Oligochaete

Tubifex tubifex

11,637

11,637

0.94

33

E

Isopod

Asellus aquaticus

29,446**

29,446

0.97

**Measured EC50 falls outside range of ICE model and a "scaled" EC50 was therefore entered in ICE model.
1: Freshwater MDR Groups

A) The family Salmonidae in the class Osteichthyes

F-16


-------
B)	A second family in the class Osteichthyes, preferably a commercially or recreationally important

warmwater species (e.g., bluegill, channel catfish, etc.)

C)	A third family in the phylum Chordata (may be in the class Osteichthyes or may be an amphibian, etc.)

D)	A planktonic crustacean (e.g., cladoceran, copepod, etc.)

E)	Abenthic crustacean (e.g., ostracod, isopod, amphipod, crayfish, etc.)

F)	An insect (e.g., mayfly, dragonfly, damselfly, stonefly, caddisfly, mosquito, midge, etc.)

G)	A family in a phylum other than Arthropoda or Chordata (e.g., Rotifera, Annelida, Mollusca, etc.)

H)	A family in any order of insect or any phylum not already represented.

Table F-6. PFHxA Final Acute Value and Protective Aquatic Acute Benchmark.

Bold values represent genera for which empirical toxicity data were available.	

Calculated Freshwater FAY based on 4 lowest values; N=33 GMAVs total





Benchmark calculated by dividing the FAY by 2 and by the carboxylic acid application factor (15.8)





GMAV









Rank

Genus

(mg/L)

ln(GMAV)

ln(GMAV)2

P=R/(N+1)

sqrt(P)

1

Brachionus

140.0

4.94

24.42

0.029

0.171

2

Amblema

191.3

5.25

27.60

0.059

0.243

3

Margaritifera

370.2

5.91

34.98

0.088

0.297

4

Megalonaias

372.3

5.92

35.04

0.118

0.343



£ (Sum):

22.03

122.04

0.29

1.05







P = cumulative probability











R = rank













N = number of GMAVs





S2 =

44.02



S = slope







L =

3.759



L = X-axis intercept





A =

5.242



A = InFAV







FAV =

189.1











FAV/2 =

94.57 mg/L (Preliminary Value)







Adjustment =

94.57 / 15.8 = 5.985 (Preliminary Value / Carboxylic Acid Application Factor)

Benchmark =

6.0 mg/L PFHxA (rounded to two significant figures)





F-17


-------
1.00 -r

o.9o
0.80 -f

0.70

s 0.60 -+
S
s

u

a

«

41

0.50
0.40

g 0.30

Oh

0.20

o.io 4

0.00
1

~

A

o

Amphibian (Empirical)
Fish (WeblCE)

Insect (WeblCE)
Invertebrate (Empirical)
Invertebrate (WeblCE)
Mollusk (WeblCE)
Preliminary Value
-Acute Benchmark

A Asellus
A Tubifex
A Crangonyx
A Limnodrilus

A Moina
+ Paratanvtarsus
O Pimephales
~ Daphnia
O Carassius
¦ Lithobates
O Lasmigona
O Gambusia
O Ortmamiiana
O Physella
A Ceriodaphnia
O Lepomis
O Lymnaea
O Ictalurus
O Oncorhynchus
A Simocephalus
O Lampsilis
A Branchinecta
A Palaemonetes
A Thamnocephalus
A Gammarus
O Danio
A Chydorus
A Macrobrachium
O Utterbackia
O Megalonaias
O Margaritifera
O Amblema
A Brachionus

—i	i	i	i	i i i 11

_i	i	i	i	i i i i i

—i	i i i	i i i 11

10	100	1,000	10,000

Genus Mean Acute Value (mg/L PFHxA)

100,000

Figure F-2. Ranked Acute PFHxA GMAVs Used for the Aquatic Life Acute Benchmark
Calculations.

F.3 Derivation of Acute Water Benchmark for Perfluorononanoic Acid (PFNA)

Quantitatively acceptable empirical data for PFNA were available for three species

comprising three genera and fulfilling two MDR groups (Table F-8). A total of 42 ICE models
were accepted for use. Of these, six ICE models were not selected for use because empirical data
were available for the genus (e.g., models for Daphnia longispina and Daphnia magna). One
other model was not used because there was an acceptable model for more closely-related
surrogate species (i.e., Daphnia magna predicting tor Pimephalespromelas). One additional
model (i.e., Daphnia magna predicting for Danio rerio) was not used because the measured EC so
values fell outside of the ICE model range and other acceptable models for taxonomically-related

F-18


-------
species with measured EC50 values within the ICE model range were used. A total of 34 ICE
models were selected for use, resulting in ICE models predicting 34 SMAVs representing 30
genera (Tables F-7 and F-8).

The combined empirical and ICE data resulted in 33 GMAVs that collectively fulfill the
eight MDR groups for deriving a freshwater criterion as defined by the Aquatic Life Criteria
Guidelines (U.S. EPA 1985). The ranked GMAVs for these combined data along with the MDR
met by each GMAV are summarized in Table F-8. GMAVs for the four most sensitive genera
were within a factor of 2.0 of each other (Table F-9). The freshwater FAV (the 5th percentile of
the genus sensitivity distribution) for PFNA is 23.18 mg/L (Table F-9), which is lower than all
of the GMAVs except for the one ICE-derived species value (the threeridge mussel, Amblema
plicata, GMAV=17.58 mg/L). The FAV was divided by two to obtain a preliminary value of
11.59 mg/L PFNA and then adjusted by the carboxylic acid application factor (15.8) to obtain
the freshwater acute water column benchmark magnitude of 0.73 mg/L PFNA (rounded to two
significant figures). This value is expected to be protective of 95% of freshwater genera exposed
to PFNA under short-term conditions of one-hour duration, if the one-hour average magnitude is
not exceeded more than once in three years (Figure F-3).

F-19


-------
Table F-7. Acceptable models for ICE-estimated Species Sensitivity to PFNA using the scaling approach for data outside of the
model bounds as in cases indicated.

Bold predicted EC50S used for SMAY calculations.			

Predicted Species

Surrogate

ICE Model Value Range
(mg/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Entered
ECso
(units)

Minimum

Maximum

Amphipod (Crangonyx
pseudogracilis)

Daphnid (Daphnia magna)

84.51A

84.51
(mg/L)

0.0008

166.8

1,035.5

(296.9-3,611.9)

Amphipod (Gammarus
fasciatus)

Daphnid (Daphnia magna)

84.51

84,510
(Hg/L)

0.0002

5,000

29.8

(10.9-81.6)

Amphipod (Gammarus
pseudolimnaeus)

Daphnid (Daphnia magna)

84.51**

84.51
(mg/L)

0.0001

68.3

102.4

(51.2-204.7)

Beaver-tail fairy shrimp
(Thamnocephalus platyurus)

Daphnid (Daphnia magna)

84.51

84,510
(Hg/L)

0.0003

8,694.5

52.1

(32.2-84.2)

Bluegill (Lepomis
macrochirus)

Daphnid (Daphnia magna)

84.51

84,510
(Ug/L)

0.0001

46,278

50.3

(33.3-75.9)

Bryozoan (Pectinatella
magnifica)

Daphnid (Daphnia magna)

84.51**

84.51
(mg/L)

0.014

0.232

648.8

(426-988.3)

Channel catfish (Ictalurus
punctatus)

African clawed frog
(Xenopus laevis)

335.8**

335.8
(mg/L)

0.018

122.5

977.6

(162.9-5,867.9)

Daphnid (Ceriodaphnia
dubia)

Daphnid (Daphnia magna)

84.51

84,510
(Hg/L)

0.0003

46,278

59.4

(38.2-92.1)

Daphnid (Ceriodaphnia
reticulata)

Daphnid (Daphnia magna)

84.51**

84.51
(mg/L)

0.0008

0.232

44.5

(16.1-123.4)

Daphnid (Daphnia galeata)*

Daphnid (Daphnia magna)

84.51**

84.51
(mg/L)

0.0001

0.646

109.2
(49.2-242.4)

Daphnid (Daphnia
longispina)*

Daphnid (Daphnia magna)

84.51**

84.51
(mg/L)

0.009

10.36

59.4
(16.2-217.7)

Daphnid (Daphnia magna)*

Water flea (Chydorus
sphaericus)

27.84

27,840
(Hg/L)

0.007

462

76.4
(48-121.5)

Daphnid (Daphnia pulex)*

Daphnid (Daphnia magna)

84.51

84,510
(Hg/L)

0.0002

4,894.7

65.8
(40.3-107.5)

F-20


-------
Predicted Species

Surrogate

ICE Model Value Range
(mg/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Entered
ECso
(units)

Minimum

Maximum

Daphnid (Daphnia pulicaria)*

Daphnid (Daphnia magna)

84.51

84,510
(Hg/L)

0.014

281.6

84.2
(18-392.8)

Daphnid (Simocephalus
serrulatus)

Daphnid (Daphnia magna)

84.51**

84.51
(mg/L)

0.0002

7.2

61.3

(33.8-111)

Daphnid (Simocephalus
vetulus)

Daphnid (Daphnia magna)

84.51A

84.51
(mg/L)

0.0001

166.8

113.7

(29.4-439.6)

Fathead minnow (Pimephales
promelas)

African clawed frog
(Xenopus laevis)

335.8**

335.8
(mg/L)

0.045

122.5

260.9

(48.9-1,393.3)

Fathead minnow (Pimephales
promelas)

Daphnid (Daphnia magna)

84.51

84,510
(Hg/L)

0.0002

46,500

67.3b
(47.1-96.2)

Fatmucket (Lampsilis
siliquoidea)

Daphnid (Daphnia magna)

84.51

84,510
(Ug/L)

0.014

8,694.5

48.6

(27.6-85.6)

Green floater (Lasmigona
subviridis)

Daphnid (Daphnia magna)

84.51A

84.51
(mg/L)

0.014

166.8

167.6

(38.9-721.5)

Isopod (Asellus aquaticus)

Daphnid (Daphnia magna)

84.51

84,510
(Ug/L)

0.0003

166.8

1,261.8

(217.7-7,314.9)

Leech (Nephelopsis obscura)

Daphnid (Daphnia magna)

84.51**

84.51
(mg/L)

0.0008

4.369

7,904.0

(1,279-48,846)

Midge (Chironomus tentans)

Daphnid (Daphnia magna)

84.51A

84.51
(mg/L)

0.0003

472

435.6

(71.7-2,647.8)

Midge (Paratanytarsus
parthenogeneticus)

Daphnid (Daphnia magna)

84.51

84,510
(Hg/L)

0.37

14,500

228.8

(101.9-513.5)

Mississippi grass shrimp
(Palaemonetes kadiakensis)

Daphnid (Daphnia magna)

84.51**

84.51
(mg/L)

0.0003

58

84.4

(33.9-210)

Neosho mucket (Lampsilis
rafinesqueana)

Daphnid (Daphnia magna)

84.51

84,510
(Hg/L)

0.042

166.8

42.9

(12.1-152.7)

Oligochaete (Limnodrilus
hoffmeisteri)

Daphnid (Daphnia magna)

84.51

84,510
(Ug/L)

0.003

281.6

181.9

(43.1-768.2)

Oligochaete (Tubifex tubifex)

Daphnid (Daphnia magna)

84.51A

84.51
(mg/L)

0.0001

4,894.7

1,225.2

(393.2-3,817.5)

F-21


-------
Predicted Species

Surrogate

ICE Model Value Range
(mg/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Entered
ECso
(units)

Minimum

Maximum

Oriental river shrimp
(Macrobrachium nipponense)

Daphnid (Daphnia magna)

84.51

84,510
(Hg/L)

0.011

281.6

65.1

(24.3-174.3)

Paper pondshell (Utterbackia
imbecillis)

Daphnid (Daphnia magna)

84.51

84,510
(Ug/L)

0.014

8,694.5

44.8

(28.6-70.3)

Pheasantshell (Ortmanniana
pectorosa)

Daphnid (Daphnia magna)

84.51

84,510
(Hg/L)

0.042

545.9

54.4

(13.3-222.4)

Rainbow trout
(Oncorhynchus mykiss)

Daphnid (Daphnia magna)

84.51

84,510
(Ug/L)

0.0001

14,500

35.4

(25.2-49.7)

Swamp lymnaea (Lymnaea
stagnalis)

Daphnid (Daphnia magna)

84.51

84,510
(Hg/L)

0.014

8,694.5

70.7

(24-208)

Tadpole physa (Physella
gyrina)

Daphnid (Daphnia magna)

84.51

84,510
(Ug/L)

0.014

8,694.5

70.6

(31.3-159.2)

Threeridge (Amblema
plicata)

Daphnid (Daphnia magna)

84.51

84,510
(Hg/L)

0.014

4,894.7

17.6

(6.1-50.8)

Vernal pool fairy shrimp
(Branchinecta lynchi)

Daphnid (Daphnia magna)

84.51

84,510
(Ug/L)

0.014

8,694.5

58.0

(28.6-117.6)

Washboard (Megalonaias
nervosa)

Daphnid (Daphnia magna)

84.51

84,510
(Hg/L)

0.014

8,694.5

32.9

(18.5-58.5)

Water flea (Chydorus
sphaericus)*

Daphnid (Daphnia magna)

84.51

84,510
(Ug/L)

0.009

977.6

30.1
(19.3-47.1)

Water flea (Moina
macrocopa)

Daphnid (Daphnia magna)

84.51

84,510
(Hg/L)

0.009

281.6

44.3

(8.8-223.1)

Western pearlshell
(Margaritifera falcata)

Daphnid (Daphnia magna)

84.51

84,510
(Ug/L)

0.014

8,694.5

40.4

(19.8-82.5)

Zebraflsh (Danio rerio)

Daphnid (Daphnia magna)

84.51**

84.51
(mg/L)

0.0001

50

504.6C
(137.4-1,853.4)

Zebraflsh-embryo (Danio
rerio-embryo)

Daphnid (Daphnia magna)

84.51

84,510
(Ug/L)

0.0001

46,500

91.9

(42.1-200.2)

*Acceptable models that were not used because genus level empirical data were available.

**Measured ECso falls outside range of ICE model and a "scaled" ECso was therefore entered in ICE model.

A |ig/L CI greater than 50-fold. Used mg/L with acceptable CI.

F-22


-------
a Saltwater surrogate species were not used.

b Not used because other more closely taxonomically-related models were available.

0 Not used because other more closely taxonomically-related models where the measured surrogate EC50 values fell within the ICE model range were available.

F-23


-------
Table F-8. Ranked PFNA Genus Mean Acute Values.

Values in bold and highlighted are derived from empirical PFNA toxicity tests with the species.

Rank

MDR

Group1

Name

Species

SMAV

GMAV

Percentile
Rank

1

G

Threeridge

Amblema plicata

17.58

17.58

0.03

2

D

Daphnid

Chydorus sphaericus

27.84

27.84

0.06

3

G

Washboard

Megalonaias nen'osa

32.90

32.90

0.09

4

A

Rainbow trout

Oncorhvnchus mvkiss

35.40

35.40

0.12

5

G

Western pearlshell

Margaritifera falcata

40.41

40.41

0.15

6

D

Water flea

Moina macrocopa

44.31

44.31

0.18

7

G

Paper pondshell

Utterbackia imbecillis

44.82

44.82

0.21

8

G

Neosho mucket

Lampsilis rafinesqueana

42.93

45.69

0.24

G

Fatmucket

Lampsilis siliquoidea

48.63

9

B

Bluegill

Lepomis macrochirus

50.29

50.29

0.26

10

D

Daphnid

Ceriodaphnia dubia

59.35

51.41

0.29

D

Daphnid

Ceriodaphnia reticulata

44.54**

11

E

Beaver-tail fairy shrimp

Thamnocephalus platvurus

52.08

52.08

0.32

12

G

Pheasantshell

Ortmanniana pectorosa

54.41

54.41

0.35

13

E

Amphipod

Gammarus fasciatus

29.82

55.26

0.38

E

Amphipod

Gammarus pseudolimnaeus

102.4**

14

E

Vernal pool fairy shrimp

Branchinecta Ivnchi

58.02

58.02

0.41

15

E

Oriental river shrimp

Macrobrachium
nipponense

65.1

65.10

0.44

16

G

Tadpole physa

Phvsella gvrina

70.62

70.62

0.47

17

G

Swamp lymnaea

Lymnaea stagnalis

70.68

70.68

0.50

18

D

Daphnid

Simocephalus serrulatus

61.28**

83.48

0.53

D

Daphnid

Simocephalus vetulus

113.7

19

E

Mississippi grass shrimp

Palaemonetes kadiakensis

84.41**

84.41

0.56

20

D

Daphnid

Daphnia magna

84.51

84.51

0.59

21

B

Zebrafish

Danio rerio

91.85

91.85

0.62

22

G

Green floater

Lasmigona subviridis

167.6

167.6

0.65

23

H

Oligochaete

Limnodrilus hoffmeisteri

181.9

181.9

0.68

24

F

Midge

Paratanytarsus
parthenogeneticus

228.8

228.8

0.71

25

B

Fathead minnow

Pimephales promelas

260.9**

260.9

0.74

26

C

Clawed frog

Xenopus sp.

335.8

335.8

0.76

27

F

Midge

Chironomus teutons

435.6

435.6

0.79

28

H

Bryozoan

Pectinate/la magnifica

648.8**

648.8

0.82

29

B

Channel catfish

Ictalurus punctatus

977.6**

977.6

0.85

30

E

Amphipod

Crangonvx pseudogracilis

1,035

1,035

0.88

31

H

Oligochaete

Tubifex tubifex

1,225

1,225

0.91

32

E

Isopod

Asellus aquaticus

1,262

1,262

0.94

33

H

Leech

Nephelopsis obscura

7,904**

7,904

0.97

**Measured EC50 falls outside range of ICE model and a "scaled" EC50 was therefore entered in ICE model.
1: Freshwater MDR Groups

A)	The family Salmonidae in the class Osteichthyes

B)	A second family in the class Osteichthyes, preferably a commercially or recreationally important

warmwater species (e.g., bluegill, channel catfish, etc.)

C)	A third family in the phylum Chordata (may be in the class Osteichthyes or may be an amphibian, etc.)

F-24


-------
D)	A planktonic crustacean (e.g., cladoceran, copepod, etc.)

E)	Abentliic crustacean (e.g., ostracod, isopod, amphipod, crayfish, etc.)

F)	An insect (e.g., mayfly, dragonfly, damselfly, stonefly, caddisfly, mosquito, midge, etc.)

G)	A family in a phylum other than Arthropoda or Chordata (e.g., Rotifera, Annelida, Mollusca, etc.)

H)	A family in any order of insect or any phylum not already represented.

Table F-9. PFNA Final Acute Value and Protective Aquatic Acute Benchmark.

Bold values represent genera for which empirical toxicity data were available.	

Calculated Freshwater FAV based on 4 lowest values; N=33 GMAVs total





Benchmark calculated by dividing the FAV by 2 and by the carboxylic acic

application factor (15.8)





GMAV









Rank

Genus

(mg/L)

ln(GMAV)

ln(GMAV)2

P=R/(N+1)

sqrt(P)

1

Amblema

17.58

2.87

8.22

0.029

0.171

2

Chydorus

27.84

3.33

11.07

0.059

0.243

3

Megalonaias

32.90

3.49

12.20

0.088

0.297

4

Oncorhynchns

35.40

3.57

12.72

0.118

0.343



£ (Sum):

13.25

44.21

0.29

1.05







P = cumulative probability











R = rank













N = number of GMAVs





S2 =

18.12



S = slope







L =

2.192



L = X-axis intercept





A =

3.143



A = InFAV







FAV =

23.18











FAV/2 =

11.59 mg/L (Preliminary Value)







Adjustment =

11.59 / 15.8 = 0.7337 (Preliminary Value / Carboxylic Acid Application Factor)

Benchmark =

0.73 mg/L PFNA (rounded to two significant figures)





F-25


-------
s

9

u

-C

a

S3

0£

<3J

1.00
0.90
0.80
0.70
0.60
0.50
0.40

P. 0.30

i.
su
Ph

0.20
0.10
0.00

¦

Amphibian (Empirical)

o

Fish (WeblCE)

+

Insect (WeblCE)

~

Invertebrate (Empirical)

A

Invertebrate (WeblCE)

O

Mollusk (WeblCE)



Preliminary Value



-Acute Benchmark

0.1

Nephelopsis A
A Asellus
A Tubifex
A Crangonvx
O Ictalurus
A Pectinatella
+ Chironomus
I Xenopus
O Pimephales
+ Paratanytarsus
A Limnodrilus
O Lasmigona
O Danio
A Daphnia
A Palaemonetes
A Simocephalus
O Lymnaea
 Physella
A Macrobrachium
A Branchinecta
A Gammarus
O Ortmanniana
A Thamnocephalus
A Ceriodaphnia
O Lepomis
O Lampsilis
O Utterbackia
A Moina
O Margaritifera
O Oncorhynchus
O Megalonaias
A Chydorus
O Amblema

10	100	1000

Genus Mean Acute Value (mg/L PFNA)

10000

Figure F-3. Ranked Acute PFNA GMAVs Used for the Aquatic Life Acute Benchmark
Calculation.

F.4 Derivation of Acute Water Benchmark for Perfluorodecanoic Acid (PFDA)

Quantitatively acceptable empirical data for PFDA were available for five species

comprising four genera and fulfilling three MDR groups (Table F-ll). A total of 85 ICE models
were accepted for use. Of these, thirteen ICE models were not selected for use because empirical
data were available for the genus (e.g., models for Chydorus sphaericus and Daphnia magna).
Four other models were not used because there were acceptable models for more closely-related
surrogate species (e.g., Daphnia magna predicting tor Pimephalespromelas). A total of 68 ICE
models were selected for use, resulting in ICE models predicting 67 SMAVs representing 56
genera (Table F-10 and F- 11)

F-26


-------
The combined empirical and ICE data resulted in 60 GMAVs that collectively fulfill the
eight MDR groups for deriving a freshwater criterion as defined by the Aquatic Life Criteria
Guidelines (U.S. EPA 1985). The ranked GMAVs for these combined data along with the MDR
met by each GMAV are summarized in Table F-ll. GMAVs for the four genera used in the
FAV calculation were within a factor of 1.3 of each other (Table F-12). The freshwater FAV
(the 5th percentile of the genus sensitivity distribution) for PFDA is 20.53 mg/L (Table F-12),
which is lower than all of the GMAVs except for three of the ICE-derived species values (the
isopod, Caecidotea brevicauda, GMAV= 8.170 mg/L the northern pike, Esox lucius,
GMAV=18.36 mg/L, and the largemouth bass, Micropterus salmoides, GMAV=19.52 mg/L).
The FAV was divided by two to obtain a preliminary value of 10.27 mg/L PFDA and then
adjusted by the carboxylic acid application factor (15.8) to obtain the freshwater acute water
column benchmark magnitude of 0.65 mg/L PFDA (rounded to two significant figures). This
value is expected to be protective of 95% of freshwater genera exposed to PFDA under short-
term conditions of one-hour duration, if the one-hour average magnitude is not exceeded more
than once in three years (Figure F-4).

F-27


-------
Table F-10. Acceptable models for ICE-estimated Species Sensitivity to PFDA using the scaling approach for data outside of
the model bounds as in cases indicated.

Predicted Species

Surrogate

ICE Model Value Range
(mg/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Entered
ECso (units)

Minimum

Maximum

Amphipod (Crangonyx
pseudogracilis)

Daphnid (Daphnia
magna)

119.7A

119.7
(mg/L)

0.0008

166.8

1,422.9

(423.1-4,784.7)

Amphipod (Gammarus
fasciatus)

Daphnid (Daphnia
magna)

119.7

119,700
(W$/L)

0.0002

5,000

39.7

(14-112.7)

Amphipod (Gammarus
pseudolimnaeus)

Daphnid (Daphnia
magna)

119.7**

119.7
(mg/L)

0.0001

68.3

140.9

(70-283.6)

Apache trout (Oncorhynchus
gilae)*

Rainbow trout
(Oncorhynchus mykiss)

32**

32
(mg/L)

0.004

1.625

20.0
(12.3-32.6)

Atlantic salmon (Salmo
salar)

Rainbow trout
(Oncorhynchus mvkiss)

32

32,000
(ug/L)

0.0002

95.86

31.0

(15.7-61.3)

Beaver-tail fairy shrimp
(Thamnocephalus platyurus)

Daphnid (Daphnia
magna)

119.7

119,700
(W$/L)

0.0003

8,694.5

71.5

(43.6-117.5)

Black bullhead (Ameiurus
melas)

Rainbow trout
(Oncorhynchus mvkiss)

32**

32
(mg/L)

<0.0001

9.810

207.8

(56.1-770)

Bluegill (Lepomis
macrochirus)

Daphnid (Daphnia
magna)

119.7

119,700
(W$/L)

0.0001

46,278

64.5b
(42-99)

Bluegill (Lepomis
macrochirus)

Rainbow trout
(Oncorhynchus mvkiss)

32

32,000
(ug/L)

0.0001

8,341.5

33.1

(27.6-39.6)

Bonytail (Gila elegans)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(kg/L)

0.003

43.07

72.7

(12.6-420.2)

Brook trout (Salvelinus
fontinalis)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(kg/L)

0.0006

96.69

31.4

(18.8-52.5)

Brown trout (Salmo trutta)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(kg/L)

0.0002

95.86

33.1

(19.8-55.3)

Bryozoan (Pectinatella
magnifica)

Daphnid (Daphnia
magna)

119.7**

119.7
(mg/L)

0.014

0.232

877.8

(553.4-1,392.2)

F-28


-------
Predicted Species

Surrogate

ICE Model Value Range
(mg/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Entered
ECso (units)

Minimum

Maximum

Bullfrog (Lithobates
catesbeianus)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(kg/L)

0.0006

13,400

58.8

(23.7-145.4)

Cape Fear shiner (Notropis
mekistocholas)

Rainbow trout
(Oncorhynchus mykiss)

32**

32
(mg/L)

0.0041

1.625

34.4

(22.8-51.9)

Channel catfish (Ictalurus
punctatus)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(kg/L)

<0.0001

13,400

31.6

(19.9-50.1)

Chinook salmon

(Oncorhynchus

tshawytscha)*

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(Mg/L)

0.003

724.4

40.0
(24-66.9)

Coho salmon (Oncorhynchus
kisutch)*

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(kg/L)

<0.0001

43.07

49.4
(38.4-63.6)

Colorado squawfish
(Ptychocheilus lucius)

Rainbow trout
(Oncorhynchus mykiss)

32A

32
(mg/L)

0.0033

43.07

195.1

(34.5-1,103.6)

Common carp (Cyprinus
carpio)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(kg/L)

0.0002

198.4

35.6

(16.3-77.7)

Cutthroat trout
(Oncorhynchus clarkii)*

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(kg/L)

<0.0001

198.4

27.1
(18.7-39.3)

Cuvier's foam froglet
(Physalaemus cuvieri)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(kg/L)

0.7099

155.5

51.8

(30.8-87.1)

Daphnid (Ceriodaphnia
dubia)

Daphnid (Daphnia
magna)

119.7

119,700
(kg/L)

0.0003

46,278

84.8

(53.8-133.7)

Daphnid (Ceriodaphnia
reticulata)

Daphnid (Daphnia
magna)

119.7**

119.7
(mg/L)

0.0008

0.232

60.4

(19.8-184.4)

Daphnid (Daphnia galeata)*

Daphnid (Daphnia
magna)

119.7**

119.7
(mg/L)

0.0001

0.646

150.2
(65.6-344.1)

Daphnid (Daphnia
longispina)*

Daphnid (Daphnia
magna)

119.7**

119.7
(mg/L)

0.009

10.36

90.7
(25.8-318.7)

Daphnid (Daphnia magna)*

Daphnid (Daphnia
pulicaria)

149.6

149,600
(kg/L)

0.009

237.4

122.1
(27.4-544.5)

Daphnid (Daphnia magna)*

Water flea (Chydorus
sphaericus)

41.13

41,130
(kg/L)

0.007

462

114.5
(70.9-184.8)

F-29


-------
Predicted Species

Surrogate

ICE Model Value Range
(mg/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Entered
ECso (units)

Minimum

Maximum

Daphnid (Daphnia pulex)*

Daphnid (Daphnia
magna)

119.7

119,700
(W$/L)

0.0002

4,894.7

93.7
(56.2-156)

Daphnid (Daphnia
pulicaria)*

Daphnid (Daphnia
magna)

119.7

119,700
(ug/L)

0.014

281.6

122.0
(24.2-614.7)

Daphnid (Simocephalus
serrulatus)

Daphnid (Daphnia
magna)

119.7**

119.7
(mg/L)

0.0002

7.2

84.9

(45.5-158.4)

Daphnid (Simocephalus
vetulus)

Daphnid (Daphnia
magna)

119.7A

119.7
(mg/L)

0.0001

166.8

147.7

(37.1-587.8)

Fathead minnow
(Pimephales promelas)

Daphnid (Daphnia
magna)

119.7

119,700
(W$/L)

0.0002

46,500

85.6b
(58.9-124.4)

Fathead minnow
(Pimephales promelas)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(ug/L)

0.0002

13,400

46.7

(36-60.7)

Fatmucket (Lampsilis
siliquoidea)

Daphnid (Daphnia
magna)

119.7

119,700
(kg/L)

0.014

8,694.5

66.6

(37-119.8)

Goldfish (Carassius auratus)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(Ug/L)

<0.0001

13,400

62.2

(31.6-122.5)

Green floater (Lasmigona
subviridis)

Daphnid (Daphnia
magna)

119.7A

119.7
(mg/L)

0.014

166.8

211.5

(51.2-874.2)

Green sunfish (Lepomis
cyanellus)

Rainbow trout
(Oncorhynchus mykiss)

32**

32
(mg/L)

<0.0001

19.93

90

(54.2-149.3)

Greenthroat darter
(Etheostoma lepidum)

Rainbow trout
(Oncorhynchus mykiss)

32**

32
(mg/L)

0.0041

1.625

34.9

(11-110.9)

Guppy (Poecilia reticulata)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(Ug/L)

0.0006

198.4

28.3

(7.6-105.3)

Isopod (Asellus aquaticus)

Daphnid (Daphnia
magna)

119.7

119,700
(kg/L)

0.0003

166.8

1,699.9

(273.7-10,559)

Isopod (Caecidotea
brevicauda)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(Ug/L)

0.0006

82

8.2

(1.9-35.7)

Lake trout (Salvelinus
namaycush)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(kg/L)

0.0002

198.4

18.0

(11.4-28.4)

F-30


-------
Predicted Species

Surrogate

ICE Model Value Range
(mg/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Entered
ECso (units)

Minimum

Maximum

Largemouth bass
(Micropterus salmoides)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(W$/L)

<0.0001

95.86

19.5

(9.5-40.3)

Leech (Nephelopsis obscura)

Daphnid (Daphnia
magna)

119.7**

119.7
(mg/L)

0.0008

4.369

11,822

(1,982-70,511)

Mayfly (Drunella grandis)

Rainbow trout
(Oncorhynchus mykiss)

32A

32
(mg/L)

0.0006

95.86

143.5

(43.1-477.3)

Medaka (Oryzias latipes)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(ug/L)

0.007

54.4

52.1

(10.9-248.7)

Midge (Chironomus tentans)

Daphnid (Daphnia
magna)

119.7A

119.7
(mg/L)

0.0003

472

611.2

(100.7-3,709.3)

Midge (Paratanytarsus
dissimilis)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(ug/L)

0.0006

1,330

105.8

(35.6-314.4)

Midge (Paratanytarsus
parthenogeneticus)

Daphnid (Daphnia
magna)

119.7

119,700
(kg/L)

0.37

14,500

321.5

(140.5-735.3)

Mississippi grass shrimp
(Palaemonetes kadiakensis)

Daphnid (Daphnia
magna)

119.7**

119.7
(mg/L)

0.0003

58

109.9

(42.7-282.7)

Mrigal carp (Cirrhinus
mrigala)

Rainbow trout
(Oncorhynchus mykiss)

32**

32
(mg/L)

0.0006

1.625

147.7

(24.4-896.1)

Neosho mucket (Lampsilis
rafinesqueana)

Daphnid (Daphnia
magna)

119.7

119,700
(Ug/L)

0.042

166.8

58.5

(15.4-222.1)

Northern leopard frog
(Lithobates pipiens)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(kg/L)

0.002

198.4

64.7

(33.7-124.2)

Northern pike (Esox lucius)

Rainbow trout
(Oncorhynchus mykiss)

32**

32
(mg/L)

0.005

1.822

18.4

(6.5-52)

Oligochaete (Limnodrilus
hoffmeisteri)

Daphnid (Daphnia
magna)

119.7

119,700
(kg/L)

0.003

281.6

232.8

(51.5-1,052.2)

Oligochaete (Tubifex
tubifex)

Daphnid (Daphnia
magna)

119.7A

119.7
(mg/L)

0.0001

4,894.7

1,672.6

(538.3-5,196.9)

Oriental river shrimp

(Macrobrachium

nipponense)

Daphnid (Daphnia
magna)

119.7

119,700
(Mg/L)

0.011

281.6

97.1

(34.6-272.8)

F-31


-------
Predicted Species

Surrogate

ICE Model Value Range
(mg/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Entered
ECso (units)

Minimum

Maximum

Paper pondshell (Utterbackia
imbecillis)

Daphnid (Daphnia
magna)

119.7

119,700
(W$/L)

0.014

8,694.5

61.0

(38.4-97.2)

Pheasantshell (Ortmanniana
pectorosa)

Daphnid (Daphnia
magna)

119.7

119,700
(ug/L)

0.042

545.9

76.0

(17.4-332.2)

Rainbow trout
(Oncorhynchus mykiss)*

Daphnid (Daphnia
magna)

119.7

119,700
(W$/L)

0.0001

14,500

45.4
(31.9-64.7)

Razorback sucker
(Xyrauchen texanus)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(ug/L)

0.0033

43.07

26.6

(5.3-134)

Riceland prawn
(Macrobrachium lanchesteri)

Rainbow trout
(Oncorhynchus mykiss)

32**

32
(mg/L)

0.003

2.04

177.6

(34.2-922.7)

Rohu (Labeo rohita)

Rainbow trout
(Oncorhynchus mykiss)

32**

32
(mg/L)

0.0006

1.625

302

(61.5-1,484.6)

Shortnose sturgeon
(Acipenser brevirostrum)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(kg/L)

0.028

95.86

45.7

(11.2-185.7)

Silver perch (Bidyanus
bidyanus)

Rainbow trout
(Oncorhynchus mykiss)

32**

32
(mg/L)

0.0014

7.075

40.3

(25.8-62.8)

Snipefly (Atherix variegata)

Rainbow trout
(Oncorhynchus mykiss)

32**

32
(mg/L)

0.0006

0.068

194.9

(89.6-424)

Sockeye salmon
(Oncorhynchus nerka)*

Rainbow trout
(Oncorhynchus mykiss)

32**

32
(mg/L)

0.028

7.5

74.6
(17.1-325.1)

Southern leopard frog
(Lithobates sphenocephalus)

Rainbow trout
(Oncorhynchus mykiss)

32**

32
(mg/L)

0.028

9.7

46.3

(19.9-107.9)

Spotfin chub (Erimonax
monachus)

Rainbow trout
(Oncorhynchus mykiss)

32**

32
(mg/L)

0.0041

1.625

24.5

(10.4-57.6)

Swamp lymnaea (Lymnaea
stagnalis)

Daphnid (Daphnia
magna)

119.7

119,700
(kg/L)

0.014

8,694.5

98.2

(32.2-299.3)

Tadpole physa (Physella
gyrina)

Daphnid (Daphnia
magna)

119.7

119,700
(Ug/L)

0.014

8,694.5

98.7

(42.7-227.9)

Threeridge (Amblema
plicata)

Daphnid (Daphnia
magna)

119.7

119,700
(kg/L)

0.014

4,894.7

24.5

(8.2-73.1)

F-32


-------
Predicted Species

Surrogate

ICE Model Value Range
(mg/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Entered
ECso (units)

Minimum

Maximum

Vernal pool fairy shrimp
(Branchinecta lynchi)

Daphnid (Daphnia
magna)

119.7

119,700
(kg/L)

0.014

8,694.506

80.6

(38.8-167.4)

Walleye (Sander vitreus)

Rainbow trout
(Oncorhynchus mykiss)

32**

32
(mg/L)

0.005

16.24

41.8

(12.1-144.6)

Washboard (Megalonaias
nervosa)

Daphnid (Daphnia
magna)

119.7

119,700
(kg/L)

0.014

8,694.5

46.0

(25.6-82.8)

Water flea (Chydorus
sphaericus)*

Daphnid (Daphnia
magna)

119.7

119,700
(Ug/L)

0.009

977.6

41.9
(26.5-66.4)

Water flea (Moina
macrocopa)

Daphnid (Daphnia
magna)

119.7

119,700
(kg/L)

0.009

281.6

56.9

(10.6-305.9)

Western pearlshell
(Margaritifera falcata)

Daphnid (Daphnia
magna)

119.7

119,700
(Ug/L)

0.014

8,694.5

54.9

(26.3-114.5)

Western toad (Anaxyrus
boreas)

Rainbow trout
(Oncorhynchus mykiss)

32**

32
(mg/L)

0.028

1.625

31.3

(4.6-211.6)

Yellow perch (Perca
flavescens)

Rainbow trout
(Oncorhynchus mykiss)

32**

32
(mg/L)

<0.0001

16.24

44.5

(21-94.5)

Zebrafish (Danio rerio)

Daphnid (Daphnia
magna)

H9 7**

l,19.7(mg/L)

0.0001

50

660. lb
(190-2,293.1)

Zebrafish (Danio rerio)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(Ug/L)

0.0002

229.1

17.6

(6.4-48.5)

Zebrafish-embryo (Danio
rerio-embryo)

Daphnid (Daphnia
magna)

119.7

119,700
(kg/L)

0.0001

46,500

115.5b
(51.4-259.4)

Zebrafish-embryo (Danio
rerio-embryo)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(Ug/L)

0.0002

8,341.5

34.4

(16.8-70.4)

*Acceptable models that were not used because genus level empirical data were available.

**Measured ECso falls outside range of ICE model and a "scaled" ECso was therefore entered in ICE model.

A |ig/L CI greater than 50-fold. Used mg/L with acceptable CI.

F-33


-------
Table F-ll. Ranked PFDA Genus Mean Acute Values.

Values in bold and highlighted are derived from empirical PFDA toxicity tests with the species.

Rank

MDR

Group1

Name

Species

SMAV

GMAV

Percentile
Rank

1

E

Isopod

Caecidotea brevicauda

8.170

8.170

0.02

2

B

Northern pike

Esox lucius

18.36**

18.36

0.03

3

B

Largemouth bass

Micropterus salmoides

19.52

19.52

0.05

4

A

Brook trout

Salvelimis fontinalis

31.44

23.80

0.07

A

Lake trout

Salvelimis namavcush

18.01

5

G

Threeridge

Amblema plicata

24.46

24.46

0.08

6

B

Spotfin chub

Erimonax monachus

24.47**

24.47

0.10

7

B

Zebrafish

Danio rerio

24.57

24.57

0.11

8

B

Razorback sucker

Xyrauchen texcmus

26.57

26.57

0.13

9

B

Guppy

Poecilia reticulata

28.31

28.31

0.15

10

C

Western toad

Anaxvrus boreas

31.29**

31.29

0.16

11

B

Channel catfish

Ictalurus punctatus

31.55

31.55

0.18

12

A

Rainbow trout

Oncorhynchus mykiss

32.00

32.00

0.20

13

A

Atlantic salmon

Salmo salar

30.96

32.02

0.21

A

Brown trout

Salmo trutta

33.12

14

B

Cape Fear shiner

Notropis mekistocholas

34.42**

34.42

0.23

15

B

Greenthroat darter

Etheostoma lepidum

34.87**

34.87

0.25

16

B

Common carp

Cyprinus carpio

35.58

35.58

0.26

17

B

Silver perch

Bidvamis bidvanus

40.28**

40.28

0.28

18

D

Daphnid

Chydorus sphaericus

41.13

41.13

0.30

19

B

Walleye

Sander vitreus

41.80**

41.80

0.31

20

B

Yellow perch

Percaflavescens

44.48**

44.48

0.33

21

B

Shortnose sturgeon

Acipenser brevirostrum

45.67

45.67

0.34

22

G

Washboard

Megalonaias nen'osa

46.01

46.01

0.36

23

B

Fathead minnow

Pimephales promelas

46.72

46.72

0.38

24

C

Cuvier's foam froglet

Phvsalaemus cuvieri

51.77

51.77

0.39

25

B

Medaka

Oryzias latipes

52.12

52.12

0.41

26

B

Green sunfish

Lepomis cyanellus

89.98**

54.56

0.43

B

Bluegill

Lepomis macrochirus

33.08

27

G

Western pearlshell

Margaritifera falcata

54.89

54.89

0.44

28

C

Bullfrog

Lithobates catesbeianus

58.75

56.04

0.46

C

Northern leopard frog

Lithobates pipiens

64.72

C

Southern leopard frog

Lithobates sphenocephalus

46.28**

29

D

Water flea

Moina macrocopa

56.93

56.93

0.48

30

G

Paper pondshell

Utterbackia imbecillis

61.04

61.04

0.49

31

B

Goldfish

Carassius auratus

62.18

62.18

0.51

32

G

Neosho mucket

Lampsilis rafinesqueana

58.54

62.42

0.52

G

Fatmucket

Lampsilis siliquoidea

66.56

33

E

Beaver-tail fairy shrimp

Thamnocephalus platvurus

71.54

71.54

0.54

34

D

Daphnid

Ceriodaphnia dubia

84.82

71.55

0.56

D

Daphnid

Ceriodaphnia reticulata

60.35**

35

B

Bonytail

Gila elegans

72.68

72.68

0.57

36

E

Amphipod

Gammarus fasciatus

39.66

74.75

0.59

E

Amphipod

Gammarus pseudolimnaeus

140.9**

F-34


-------
Rank

MDR

Group1

Name

Species

SMAV

GMAV

Percentile
Rank

37

G

Pheasantshell

Ortmanniana pectorosa

76.02

76.02

0.61

38

C

Clawed frog

Xenopus sp.

76.50

76.50

0.62

39

E

Vernal pool fairy shrimp

Brcmchinecta Ivnchi

80.58

80.58

0.64

40

G

Swamp lymnaea

Lymnaea stagnalis

98.19

98.19

0.66

41

G

Tadpole physa

Phvsella gvrina

98.69

98.69

0.67

42

E

Mississippi grass shrimp

Palaemonetes kadiakensis

109.9**

109.9

0.69

43

D

Daphnid

Simocephalus serrulatus

84.92**

112.0

0.70

D

Daphnid

Simocephalus vetulus

147.7

44

E

Riceland prawn

Macrobrachium
lanchesteri

177.6**

131.3

0.72

E

Oriental river shrimp

Macrobrachium
nipponense

97.09

45

F

Mayfly

Drunella grandis

143.5

143.5

0.74

46

D

Daphnid

Daphnia magna

145.5

147.5

0.75

D

Daphnid

Daphnia pulicaria

149.6

47

B

Mrigal carp

Cirrhinus mrigala

147 7**

147.7

0.77

48

F

Midge

Paratanvtarsus dissimilis

105.8

184.4

0.79

F

Midge

Paratanytarsus
parthenogeneticus

321.5

49

F

Snipefly

Atherix variegata

194 9**

194.9

0.80

50

B

Colorado squawfish

Ptvchocheilus lucius

195.1

195.1

0.82

51

B

Black bullhead

Ameiurus melas

207.8**

207.8

0.84

52

G

Green floater

Lasmigona subviridis

211.5

211.5

0.85

53

H

Oligochaete

Limnodrilus hoffmeisteri

232.8

232.8

0.87

54

B

Rohu

Labeo rohita

302.0**

302.0

0.89

55

F

Midge

Chironomus teutons

611.2

611.2

0.90

56

H

Bryozoan

Pectinate/la magnifica

877.8**

877.8

0.92

57

E

Amphipod

Crangonvx pseudogracilis

1,423

1,423

0.93

58

H

Oligochaete

Tubifex tubifex

1,673

1,673

0.95

59

E

Isopod

Asellus aquaticus

1,700

1,700

0.97

60

H

Leech

Nephelopsis obscura

11,822**

11,822

0.98

**Measured EC50 falls outside range of ICE model and a "scaled" EC50 was therefore entered in ICE model.

1: Freshwater MDR Groups

A)	The family Salmonidae in the class Osteichthyes

B)	A second family in the class Osteichthyes, preferably a commercially or recreationally important

warmwater species (e.g., bluegill, channel catfish, etc.)

C)	A third family in the phylum Chordata (may be in the class Osteichthyes or may be an amphibian, etc.)

D)	A planktonic crustacean (e.g., cladoceran, copepod, etc.)

E)	Abenthic crustacean (e.g., ostracod, isopod, amphipod, crayfish, etc.)

F)	An insect (e.g., mayfly, dragonfly, damselfly, stonefly, caddisfly, mosquito, midge, etc.)

G)	A family in a phylum other than Arthropoda or Chordata (e.g., Rotifera, Annelida, Mollusca, etc.)

H)	A family in any order of insect or any phylum not already represented.

F-35


-------
Table F-12. PFDA Final Acute Value and Protective Aquatic Acute Benchmark

Calculated Freshwater FAV based on 4 values closest to 0.05 percentile; N=60 GMAVs total

Benchmark calculated by dividing the FAV by 2 and by the carboxylic acid application factor (15.8)





GMAV









Rank

Genus

(mg/L)

ln(GMAV)

ln(GMAV)2

P=R/(N+1)

sqrt(P)

2

Esox

18.36

2.91

8.47

0.033

0.181

3

Micropterus

19.52

2.97

8.83

0.049

0.222

4

Salve limis

23.80

3.17

10.05

0.066

0.256

5

Amblema

24.46

3.20

10.22

0.082

0.286



£ (Sum):

12.25

37.57

0.23

0.95







P = cumulative probability











R = rank













N = number of GMAVs





S2 =

9.92



S = slope







L =

2.318



L = X-axis intercept





A =

3.022



A = InFAV







FAV =

20.53











FAV/2 =

10.27 mg/L (Preliminary Value)







Adjustment =

10.27 / 15.8 = 0.6498 (Preliminary Value / Carboxylic Acid Application Factor)

Benchmark =

0.65 mg/L PFDA (rounded to two significant figures)





F-36


-------
a

©

SJ

cs

;>

s

s

u
G

£ 0.40 4
—

| 0.30 4
s

Oh

^ Asellus

A Nephelopsis

O Labeo
k Lin

	 Tubifex

A Crangonyx
A Pectinatella
+ Chironomus

Carassius
Utterbackia
Moina
Lithobates
Margaritifera
Lepomis
Oryzias
. Physalaemus
Pimephales
Megalonaias
Acipenser
Perca
Sander
Chydorus
, Bidyanus
Cvprinus
Etheo stoma
Notropis
Salmo
Oncorhynchus
Ictalums
. Anaxyrus
Poecilia
Xyrauchen
Dahio
Erimonax
Amblema
Salvelinus

A Limnodnlus
Lasmigona
O Amehmis
Q Ptychodieilus
+ Atherix
+ Paratanytarsus
Q Cirrhinus
A Daphnia
-r Dninella
A Macrobraclium
A Simocephalus
A Palaemonetes
Phy sella
O Lynmaea
Brarichinecta
Xenopus
Ortmanmana
Gammarus
Gila

Cerio daphnia
Thamno cephalus
Lamps ills

O Micropterus
O Esox

¦

Amphibian (Empirical)

~

Amphibian (WeblCE)

•

Fish (Empirical)

o

Fish (WeblCE)

+

Insect (WeblCE)

~

Invertebrate (Empirical)

A

Invertebrate (WeblCE)

O

Mollusk (WeblCE)



• Preliminary Value



-Acute Benchmark

10	100	1000	10000

Genus Mean Acute Value (mg/L PFDA)

100000

Figure F-4. Ranked Acute PFDA GMAVs Used for the Aquatic Life Acute Benchmark
Calculation.

F.5 Derivation of Acute Water Benchmark for Perfluorobutanesulfonic Acid (PFBS)

Quantitatively acceptable empirical data for PFBS were available for four species

comprising four genera and fulfilling two MDR groups (Table F-14). A total of 119 ICE models
were accepted for use. Of these, seventeen ICE models were not selected for use because
empirical data were available for the genera (e.g., models for Lepomis macrochirus and Daphnia
magna). Seven ICE models were not selected for use because they were derived based on
saltwater species surrogates (e.g., Americamysis bahia predicting for Gammams fasciatus).
Sixteen other models were not used because there were acceptable models for more closely-
related surrogate species (e.g., Lepomis macrochirus predicting for Cyprinus carpio). Two

F-37


-------
additional models (e.g., Ictaluruspunctatus predicting for Lithobates catesbeianus) were not
used because the measured EC50 values fell outside of the ICE model range and other acceptable
models for taxonomically-related species with measured EC50 values within the ICE model range
were used. A total of 77 ICE models were selected for use, resulting in ICE models predicting 66
SMAVs, representing 54 genera (Tables F-13 and F- 14)

The combined empirical and ICE data resulted in 58 GMAVs that collectively fulfill the
eight MDR groups for deriving a freshwater criterion as defined by the Aquatic Life Criteria
Guidelines (U.S. EPA 1985). The ranked GMAVs for these combined data along with the MDR
met by each GMAV are summarized in Table F-14. GMAVs for the four most sensitive genera
were within a factor of 1.9 of each other (Table F-15). The freshwater FAV (the 5th percentile of
the genus sensitivity distribution) for PFBS is 646.3 mg/L (Table F-15), which is lower than all
of the GMAVs except for two of the ICE-derived species values (the threeridge mussel,
Amblemaplicata, GMAV=383.6 mg/L and the greenthroat darter, Etheostoma lepidum,
GMAV=518.0 mg/L). The FAV was divided by two to obtain a preliminary benchmark value of
323.1 mg/L PFBS and then adjusted by the sulfonic acid application factor (42.3) to obtain the
freshwater acute water column benchmark magnitude of 7.6 mg/L PFBS (rounded to two
significant figures). This value is expected to be protective of 95% of freshwater genera exposed
to PFBS under short-term conditions of one-hour duration, if the one-hour average magnitude is
not exceeded more than once in three years (Figure F-5).

F-38


-------
Table F-13. Acceptable models for ICE-estimated Species Sensitivity to PFBS using the scaling approach for data outside of
the model bounds as in cases indicated.

Bold predicted EC50S used for SMAY calculations.			

Predicted Species

Surrogate

ICE Model Value Range
(mg/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Entered
ECso
(units)

Minimum

Maximum

African clawed frog
(Xenopus laevis)

Fathead minnow
(Pimephales promelas)

1,938**

1,938
(mg/L)

0.018

140.2

1,868.8

(685.1-5,097.5)

Amphipod (Crangonyx
pseudogracilis)

Daphnid (Daphnia magna)

2,183**

2,183
(mg/L)

0.0008

166.8

20,152

(5,302.8-76,585)

Amphipod (Gammarus
fasciatus)

Daphnid (Daphnia magna)

2,183

2,183,000
(Ug/L)

0.0002

5,000

428.1

(107.9-1,697.9)

Amphipod (Gammarus
fasciatus)

Mysid (Americamysis
bahia)

372**

372
(mg/L)

<0.0001

4.4

826.2a
(280.5-2,433.7)

Amphipod (Gammarus
minus)

Fathead minnow
(Pimephales promelas)

1,938**

1,938
(mg/L)

0.239

107.6

7,872.6

(1,934.1-32,046)

Amphipod (Gammarus
pseudolimnaeus)

Daphnid (Daphnia magna)

2,183**

2,183
(mg/L)

0.0001

68.3

2,010.4

(770.9-5,242.9)

Amphipod (Gammarus
pseudolimnaeus)

Mysid (Americamysis
bahia)

372**

372
(mg/L)

<0.0001

7.17

220.9a
(87-560.8)

Amphipod (Hyalella azteca)

Mysid (Americamysis
bahia)

372**

372
(mg/L)

<0.0001

193.4

251.T
(156.5-817.5)

Arctic grayling (Thymallus
arcticus)

Bluegill (Lepomis
macrochirus)

6,452**

6,452
(mg/L)

0.038

150.8

5,722.4

(958.9-34,148)

Atlantic salmon (Salmo salar)

Bluegill (Lepomis
macrochirus)

6,452**

6,452
(mg/L)

0.004

123.3

5,330.1

(2,929.8-9,696.8)

Atlantic salmon (Salmo salar)

Fathead minnow
(Pimephales promelas)

1,938**

1,938
(mg/L)

0.044

140.2

493.4

(76.8-3,169.5)

Beaver-tail fairy shrimp
(Thamnocephalus platyurus)

Daphnid (Daphnia magna)

2,183

2,183,000
(Hg/L)

0.0003

8,694.5

1,010.1

(525.1-1,942.9)

Black bullhead (Ameiurus
melas)

Fathead minnow
(Pimephales promelas)

1,938**

1,938
(mg/L)

0.001

20.92

2,628.4

(591-11,689)

F-39


-------
Predicted Species

Surrogate

ICE Model Value Range
(mg/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Entered
ECso
(units)

Minimum

Maximum

Bluegill (Lepomis
macrochirus)*

Daphnid (Daphnia magna)

2,183

2,183,000
(Hg/L)

0.0001

46,278

512.5
(285.3-920.4)

Bluegill (Lepomis
macrochirus)*

Fathead minnow
(Pimephales promelas)

1,938

1,938,000
(Ug/L)

0.0002

50,154

751.0
(390-1,446)

Bluegill (Lepomis
macrochirus)*

Mysid (Americamysis
bahia)

372**

372
(mg/L)

<0.0001

79

1,574.6
(1,174.5-2,111)

Bonytail (Gila elegans)

Bluegill (Lepomis
macrochirus)

6,452**

6,452
(mg/L)

0.054

150.8

1,895,4b
(374.7-9,588.1)

Bonytail (Gila elegans)

Fathead minnow
(Pimephales promelas)

1,938**

1,938
(mg/L)

0.058

8.32

880.6

(168.2-4,610.9)

Brook trout (Salvelinus
fontinalis)

Bluegill (Lepomis
macrochirus)

6,452**

6,452
(mg/L)

0.0004

123.3

4,301.9

(2,141.8-8,640.5)

Brook trout (Salvelinus
fontinalis)

Fathead minnow
(Pimephales promelas)

1,938**

1,938
(mg/L)

0.001

140.2

506.7

(192.5-1,334)

Brown trout (Salmo trutta)

Bluegill (Lepomis
macrochirus)

6,452**

6,452
(mg/L)

0.002

123.3

2,390.2

(732.9-7,795.5)

Bullfrog (Lithobates
catesbeianus)

Bluegill (Lepomis
macrochirus)

6,452**

6,452
(mg/L)

0.0004

201

7,340c
(3,632.2-14,833)

Bullfrog (Lithobates
catesbeianus)

Fathead minnow
(Pimephales promelas)

1,938

1,938,000
(Ug/L)

0.001

26,500

1,791.4

(756.1-4,244.6)

Catla (Gibelion catla)

Fathead minnow
(Pimephales promelas)

1,938**

1,938
(mg/L)

0.333

8.32

1,718.3

(810.4-3,643.3)

Channel catfish (Ictalurus
punctatus)

Bluegill (Lepomis
macrochirus)

6,452**

6,452
(mg/L)

0.0004

517.8

6,851.9C
(4,607-10,191)

Channel catfish (Ictalurus
punctatus)

Fathead minnow
(Pimephales promelas)

1,938

1,938,000
(Hg/L)

0.001

26,500

1,617.3

(755.1-3,464.4)

Chinook salmon
(Oncorhynchus tshawytscha)

Bluegill (Lepomis
macrochirus)

6,452**

6,452
(mg/L)

0.004

150.8

2,417.4

(631.8-9,249.7)

Chinook salmon
(Oncorhynchus tshawytscha)

Fathead minnow
(Pimephales promelas)

1,938**

1,938
(mg/L)

0.011

140.2

397.2

(94.8-1,664.3)

F-40


-------
Predicted Species

Surrogate

ICE Model Value Range
(mg/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Entered
ECso
(units)

Minimum

Maximum

Coho salmon (Oncorhynchus
kisutch)

Bluegill (Lepomis
macrochirus)

6,452**

6,452
(mg/L)

0.0004

150.8

3,010.5

(1,399.5-6,475.7)

Coho salmon (Oncorhynchus
kisutch)

Fathead minnow
(Pimephales promelas)

1,938**

1,938
(mg/L)

0.001

20.92

589.1

(232.4-1,493.1)

Colorado squawfish
(Ptychocheilus lucius)

Fathead minnow
(Pimephales promelas)

1,938**

1,938
(mg/L)

0.058

20.92

868.0

(215.4-3,498.7)

Common carp (Cyprinus
carpio)

Bluegill (Lepomis
macrochirus)

6,452**

6,452
(mg/L)

0.0009

180

7,708.8b
(3,816-15,573)

Common carp (Cyprinus
carpio)

Fathead minnow
(Pimephales promelas)

1,938**

1,938
(mg/L)

0.005

133

2,074.7

(1,359.3-3,166.5)

Cutthroat trout
(Oncorhynchus clarkii)

Bluegill (Lepomis
macrochirus)

6,452**

6,452
(mg/L)

0.0004

180

5,874.5

(3,202.5-10,776)

Cutthroat trout
(Oncorhynchus clarkii)

Fathead minnow
(Pimephales promelas)

1,938**

1,938
(mg/L)

0.001

140.2

645.0

(354.7-1,172.7)

Daphnid (Ceriodaphnia
dubia)

Daphnid (Daphnia magna)

2,183

2,183,000
(Ug/L)

0.0003

46,278

1,666.6

(909-3,055.6)

Daphnid (Ceriodaphnia
dubia)

Mysid (Americamysis
bahia)

372

372,000
(Hg/L)

<0.0001

4,100

171.7a
(36.6-804.9)

Daphnid (Daphnia
longispina)*

Daphnid (Daphnia magna)

2,183**

2,183
(mg/L)

0.009

10.36

3,094.2
(706.1-13,559)

Daphnid (Daphnia magna)*

Mysid (Americamysis
bahia)

372

372,000
(Hg/L)

<0.0001

11,000

243.9
(143.9-413.4)

Daphnid (Daphnia pulex)*

Daphnid (Daphnia magna)

2,183

2,183,000
(Ug/L)

0.0002

4,894.7

1,768.1
(893.9-3,497.1)

Daphnid (Daphnia pulicaria)*

Daphnid (Daphnia magna)

2,183**

2,183
(mg/L)

0.014

281.6

1,712.1
(445.5-6,580.5)

Daphnid (Simocephalus
serrulatus)

Daphnid (Daphnia magna)

2,183**

2,183
(mg/L)

0.0002

7.2

1,291.0

(482.8-3,451.9)

Daphnid (Simocephalus
vetulus)

Daphnid (Daphnia magna)

2,183**

2,183
(mg/L)

0.0001

166.8

1,307.9

(202.3-8,457.1)

F-41


-------
Predicted Species

Surrogate

ICE Model Value Range
(mg/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Entered
ECso
(units)

Minimum

Maximum

Fathead minnow (Pimephales
promelas)*

Bluegill (Lepomis
macrochirus)

6,452

6,452,000
(Hg/L)

0.0001

27,540

3,399.2
(1,631.9-7,080.3)

Fathead minnow (Pimephales
promelas)*

Daphnid (Daphnia magna)

2,183

2,183,000
(Ug/L)

0.0002

46,500

639.5
(375.7-1,088.5)

Fathead minnow (Pimephales
promelas)*

Mysid (Americamysis
bahia)

372

372,000
(Hg/L)

<0.0001

11,000

158.6
(74.6-337.5)

Fatmucket (Lampsilis
siliquoidea)

Daphnid (Daphnia magna)

2,183

2,183,000
(Ug/L)

0.014

8,694.5

912.5

(399-2,086.8)

Fatmucket (Lampsilis
siliquoidea)

Fathead minnow
(Pimephales promelas)

1,938A

1,938
(mg/L)

0.013

10,831

487.4b
(125.7-1,889.9)

Flagfish (Jordanella floridae)

Bluegill (Lepomis
macrochirus)

6,452**

6,452
(mg/L)

0.0004

113

5,648.7

(1,648.7-19,354)

Flagfish (Jordanella floridae)

Fathead minnow
(Pimephales promelas)

1,938**

1,938
(mg/L)

0.001

32.54

1,232.7

(388.5-3,911.1)

Goldfish (Carassius auratus)

Bluegill (Lepomis
macrochirus)

6,452**

6,452
(mg/L)

0.0004

201

8,832.2b
(4,286.5-18,199)

Goldfish (Carassius auratus)

Fathead minnow
(Pimephales promelas)

1,938

1,938,000
(Hg/L)

0.001

26,500

2,152.4

(1,075.2-4,309)

Green floater (Lasmigona
subviridis)

Daphnid (Daphnia magna)

2,183**

2,183
(mg/L)

0.014

166.8

1,471.9

(318.2-6,807.8)

Green sunfish (Lepomis
cyanellus)*

Bluegill (Lepomis
macrochirus)

6,452**

6,452
(mg/L)

0.004

113

7,710.8
(3,925.7-15,146)

Green sunfish (Lepomis
cyanellus)*

Fathead minnow
(Pimephales promelas)

1,938**

1,938
(mg/L)

0.011

26.69

720.9
(107.3-4,841.9)

Greenthroat darter
(Etheostoma lepidum)

Fathead minnow
(Pimephales promelas)

1,938**

1,938
(mg/L)

0.006

8.32

518.0

(105.9-2,533.1)

Guppy (Poecilia reticulata)

Bluegill (Lepomis
macrochirus)

6,452**

6,452
(mg/L)

0.0004

180

5,295.7

(2,358.2-11,892)

Guppy (Poecilia reticulata)

Fathead minnow
(Pimephales promelas)

1,938**

1,938
(mg/L)

0.001

140.2

1,407.4

(816.4-2,426.3)

F-42


-------
Predicted Species

Surrogate

ICE Model Value Range
(mg/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Entered
ECso
(units)

Minimum

Maximum

Indian bullfrog (Euphlyctis
hexadactylus)

Bluegill (Lepomis
macrochirus)

6,452**

6,452
(mg/L)

0.038

113

2,330.8

(410.6-13,232)

Isopod (Asellus aquaticus)

Bluegill (Lepomis
macrochirus)

6,452**

6,452
(mg/L)

0.003

123.3

9,389.6b
(2,238.7-39,383)

Isopod (Asellus aquaticus)

Daphnid (Daphnia magna)

2,183**

2,183
(mg/L)

0.0003

166.8

55,185

(17,354-175,483)

Isopod (Asellus aquaticus)

Mysid (Americamysis
bahia)

372**

372
(mg/L)

<0.0001

6.983

5,240.6a
(964.7-28,468.1)

Isopod (Caecidotea
brevicauda)

Bluegill (Lepomis
macrochirus)

6,452**

6,452
(mg/L)

0.0004

26.7

3,753.7

(1,068.9-13,183)

Isopod (Caecidotea
brevicauda)

Fathead minnow
(Pimephales promelas)

1,938**

1,938
(mg/L)

0.001

20.92

286.2

(83.9-976.8)

Lake trout (Salvelinus
namaycush)

Bluegill (Lepomis
macrochirus)

6,452**

6,452
(mg/L)

0.003

180

2,719.2

(1,374-5,381.5)

Lake trout (Salvelinus
namavcush)

Fathead minnow
(Pimephales promelas)

1,938**

1,938
(mg/L)

0.044

133

374.7

(171.7-817.4)

Largemouth bass
(Micropterus salmoides)

Bluegill (Lepomis
macrochirus)

6,452**

6,452
(mg/L)

0.0004

123.3

6,684.3

(4,375-10,213)

Medaka (Oryzias latipes)

Fathead minnow
(Pimephales promelas)

1,938**

1,938
(mg/L)

0.003

1,420

2,670.2

(1,679.9-4,244.3)

Midge (Chironomus
plumosus)

Mysid (Americamysis
bahia)

372**

372
(mg/L)

<0.0001

87.6

672. la
(228.5-1,977.3)

Midge (Chironomus tentans)

Bluegill (Lepomis
macrochirus)

6,452**

6,452
(mg/L)

0.0001

517.8

10,675

(1,696.1-67,189)

Midge (Paratanytarsus
dissimilis)

Bluegill (Lepomis
macrochirus)

6,452**

6,452
(mg/L)

0.0004

201

18,234

(3,711.6-89,583)

Midge (Paratanytarsus
dissimilis)

Fathead minnow
(Pimephales promelas)

1,938**

1,938
(mg/L)

0.001

1,430

6,295.4

(1,795.6-22,072)

Midge (Paratanytarsus
parthenogeneticus)

Daphnid (Daphnia magna)

2,183

2,183,000
(Hg/L)

0.37

14,500

5,481.6

(1,674.6-17,944)

F-43


-------
Predicted Species

Surrogate

ICE Model Value Range
(mg/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Entered
ECso
(units)

Minimum

Maximum

Midge (Paratanytarsus
parthenogeneticus)

Fathead minnow
(Pimephales promelas)

1,938A

1,938
(mg/L)

0.28

10,600

6,562.8b
(1,034.9-41,616)

Mississippi grass shrimp
(Palaemonetes kadiakensis)

Daphnid (Daphnia magna)

2,183**

2,183
(mg/L)

0.0003

58

993.2

(233.2-4,230.7)

Mosquitofish (Gambusia
affinis)

Fathead minnow
(Pimephales promelas)

1,938A

1,938
(mg/L)

0.006

26,500

1,349.6

(216.3-8,421.7)

Mozambique tilapia
(Oreochromis mossambicus)

Fathead minnow
(Pimephales promelas)

1,938**

1,938
(mg/L)

0.044

140.2

1,578

(636.6-3,911.6)

Mrigal carp (Cirrhinus
mrigala)

Fathead minnow
(Pimephales promelas)

1,938**

1,938
(mg/L)

0.1969

8.32

1,132.5

(742.8-1,726.6)

Neosho mucket (Lampsilis
rafinesqueana)

Daphnid (Daphnia magna)

2,183**

2,183
(mg/L)

0.042

166.8

1,653.6

(781.8-3,497.4)

Nile tilapia (Oreochromis
niloticus)

Bluegill (Lepomis
macrochirus)

6,452**

6,452
(mg/L)

0.002

6.969

8,705.2

(7,662.2-9,890.2)

Northern leopard frog
(Lithobates pipiens)

Bluegill (Lepomis
macrochirus)

6,452**

6,452
(mg/L)

0.008

180

14,976.0

(2,672.6-83,915)

Oligochaete (Limnodrilus
hoffmeisteri)

Daphnid (Daphnia magna)

2,183**

2,183
(mg/L)

0.003

281.6

13,646.0

(5,444.3-34,203)

Oligochaete (Lumbriculus
variegatus)

Fathead minnow
(Pimephales promelas)

1,938**

1,938
(mg/L)

0.001

140.2

2,368.1

(1,042.6-5,378.4)

Oligochaete (Tubifex tubifex)

Bluegill (Lepomis
macrochirus)

6,452**

6,452
(mg/L)

0.0001

113

2,696.5b
(423.5-17,169)

Oligochaete (Tubifex tubifex)

Daphnid (Daphnia magna)

2,183A

2,183
(mg/L)

0.0001

4,894.7

22,428.0

(5,594.7-89,907)

Oligochaete (Tubifex tubifex)

Fathead minnow
(Pimephales promelas)

1,938A

1,938
(mg/L)

0.015

10,831

2,039.0b
(591.9-7,024)

Oriental river shrimp
(Macrobrachium nipponense)

Daphnid (Daphnia magna)

2,183**

2,183
(mg/L)

0.011

281.6

978.3

(434.4-2,203.5)

Paper pondshell (Utterbackia
imbecillis)

Daphnid (Daphnia magna)

2,183

2,183,000
(Hg/L)

0.014

8,694.5

802.5

(422.2-1,525.5)

F-44


-------
Predicted Species

Surrogate

ICE Model Value Range
(mg/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Entered
ECso
(units)

Minimum

Maximum

Paper pondshell (Utterbackia
imbecillis)

Fathead minnow
(Pimephales promelas)

1,938A

1,938
(mg/L)

0.134

10,831

478.6b
(99.6-2,299.1)

Peppered loach
(Lepidocephalichthys guntea)

Fathead minnow
(Pimephales promelas)

1,938**

1,938
(mg/L)

0.1969

43.79

1,937.8

(699.6-5,367.4)

Pheasantshell (Ortmanniana
pectorosa)

Daphnid (Daphnia magna)

2,183**

2,183
(mg/L)

0.042

545.9

1,625.4

(540-4,892.6)

Rainbow trout
(Oncorhynchus mykiss)

Bluegill (Lepomis
macrochirus)

6,452**

6,452,000
(Ug/L)

0.0001

7,100

3,208.6

(2,275.7-4,524)

Rainbow trout
(Oncorhynchus mykiss)

Daphnid (Daphnia magna)

2,183

2,183,000
(Hg/L)

0.0001

14,500

361.9b
(221.2-592.1)

Rainbow trout
(Oncorhynchus mykiss)

Fathead minnow
(Pimephales promelas)

1,938

1,938,000
(Ug/L)

0.0002

26,500

788.9

(505.7-1,230.8)

Rainbow trout
(Oncorhynchus mykiss)

Mysid (Americamysis
bahia)

372**

372
(mg/L)

<0.0001

113

l,127.7a
(854.1-1,489)

Razorback sucker
(Xyrauchen texanus)

Bluegill (Lepomis
macrochirus)

6,452**

6,452
(mg/L)

0.054

150.8

l,223b
(295.2-5,067.6)

Razorback sucker
(Xyrauchen texanus)

Fathead minnow
(Pimephales promelas)

1,938**

1,938
(mg/L)

0.058

8.32

742.7

(170.7-3,231.1)

Rohu (Labeo rohita)

Bluegill (Lepomis
macrochirus)

6,452**

6,452
(mg/L)

0.0008

7.326

22,073b
(3,315.4-146,959)

Rohu (Labeo rohita)

Fathead minnow
(Pimephales promelas)

1,938**

1,938
(mg/L)

0.002

8.32

3,518.1

(750.6-16,489)

Swamp lymnaea (Lymnaea
stagnalis)

Daphnid (Daphnia magna)

2,183

2,183,000
(Ug/L)

0.014

8,694.5

1,523.7

(337.4-6,881.2)

Tadpole physa (Physella
gyrina)

Daphnid (Daphnia magna)

2,183

2,183,000
(Hg/L)

0.014

8,694.5

1,608.6

(512.8-5,046.1)

Tadpole physa (Physella
gyrina)

Fathead minnow
(Pimephales promelas)

1,938A

1,938
(mg/L)

0.814

10,831

263,4b
(41-1,694.5)

Threeridge (Amblema
plicata)

Daphnid (Daphnia magna)

2,183

2,183,000
(Hg/L)

0.014

4,894.7

383.6

(85.7-1,717.6)

F-45


-------
Predicted Species

Surrogate

ICE Model Value Range
(mg/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Entered
ECso
(units)

Minimum

Maximum

Threeridge (Amblema
plicata)

Fathead minnow
(Pimephales promelas)

1,938

1,938,000
(Hg/L)

0.814

10,831

250.3b
(39.4-1,592.2)

Vernal pool fairy shrimp
(Branchinecta lynchi)

Daphnid (Daphnia magna)

2,183

2,183,000
(Ug/L)

0.014

8,694.5

1,246.4

(457-3,399.4)

Walleye (Sander vitreus)

Bluegill (Lepomis
macrochirus)

6,452**

6,452
(mg/L)

0.004

6.722

2,272.6

(535.8-9,639.4)

Washboard (Megalonaias
nervosa)

Daphnid (Daphnia magna)

2,183

2,183,000
(Ug/L)

0.014

8,694.5

755.1

(345.4-1,650.5)

Water flea (Chydorus
sphaericus)

Daphnid (Daphnia magna)

2,183**

2,183
(mg/L)

0.009

977.6

934.5

(569.8-1,532.6)

Water flea (Moina
macrocopa)

Daphnid (Daphnia magna)

2,183**

2,183
(mg/L)

0.009

281.6

3,183.1

(1,038.3-9,758.3)

Western pearlshell
(Margaritifera falcata)

Daphnid (Daphnia magna)

2,183

2,183,000
(Hg/L)

0.014

8,694.5

706.1

(255.8-1,948.9)

White sucker (Catostomus
commersonii)

Bluegill (Lepomis
macrochirus)

6,452**

6,452
(mg/L)

0.035

123.3

4,920.4b
(2,000.4-12,103)

White sucker (Catostomus
commersonii)

Fathead minnow
(Pimephales promelas)

1,938**

1,938
(mg/L)

0.044

140.2

2,552.9

(896.1-7,273)

Yellow perch (Perca
flavescens)

Bluegill (Lepomis
macrochirus)

6,452**

6,452
(mg/L)

0.0004

9.41

4,050.0

(2,122-7,729.6)

Yellow perch (Perca
flavescens)

Fathead minnow
(Pimephales promelas)

1,938**

1,938
(mg/L)

0.001

20.92

504.lb
(168.3-1,509.3)

Zebrafish (Danio rerio)*

Daphnid (Daphnia magna)

2,183**

2,183
(mg/L)

0.0001

50

6,202.4
(1,930.8-19,924)

Zebrafish (Danio rerio)*

Fathead minnow
(Pimephales promelas)

1,938**

1,938
(mg/L)

0.013

26.69

2,005.9
(1,169.6-3,440.3)

Zebrafish (Danio rerio)*

Mysid (Americamysis
bahia)

372**

372
(mg/L)

<0.0001

5.666

6,109.6
(1,057.2-35,308)

Zebrafish-embryo (Danio
rerio-embryo)*

Daphnid (Daphnia magna)

2,183

2,183,000
(Hg/L)

0.0001

46,500

780.8
(257.2-2,370.6)

F-46


-------
Predicted Species

Surrogate

ICE Model Value Range
(mg/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Entered
ECso
(units)

Minimum

Maximum

Zebrafish-embryo (Danio
rerio-embryo)*

Fathead minnow
(Pimephales promelas)

1,938

1,938,000
(Hg/L)

0.004

70,200

1,707.9
(1,061.5-2,748)

* Acceptable models that were not used because genus level empirical data were available.

**Measured ECso falls outside range of ICE model and a "scaled" ECso was therefore entered in ICE model.

A |ig/L CI greater than 50-fold. Used mg/L with acceptable CI.
a Saltwater surrogate species were not used.

b Not used because other more closely taxonomically-related models were available.

0 Not used because other more closely taxonomically-related models where the measured surrogate ECso values fell within the ICE model range were available.

F-47


-------
Table F-14. Ranked PFBS Genus Mean Acute Values.

Values in bold and highlighted are derived from empirical PFBS toxicity tests with the species.

Rank

MDR

Group1

Name

Species

SMAV

GMAV

Percentile
Rank

1

G

Threeridge

Amblema plicata

383.6

383.6

0.02

2

B

Greenthroat darter

Etheostoma lepidum

518.0**

518.0

0.03

3

G

Western pearlshell

Margaritifera falcata

706.1

706.1

0.05

4

B

Razorback sucker

Xvrauchen texanus

742.7**

742.7

0.07

5

G

Washboard

Megalonaias nen'osa

755.1

755.1

0.08

6

G

Paper pondshell

Utterbackia imbecillis

802.5

802.5

0.10

7

B

Colorado squawfish

Ptvchocheilus lucius

868.0**

868.0

0.12

8

B

Bonytail

Gila elegans

880.6**

880.6

0.14

9

D

Water flea

Chydorus sphaericus

934.5**

934.5

0.15

10

E

Oriental river shrimp

Macrobrachium
nipponense

978.3**

978.3

0.17

11

E

Mississippi grass shrimp

Palaemonetes kadiakensis

993.2**

993.2

0.19

12

E

Beaver-tail fairy shrimp

Thamnocephalus platvurus

1,010

1,010

0.20

13

E

Isopod

Caecidotea brevicauda

1,036**

1,036

0.22

14

B

Mrigal carp

Cirrhinus mrigala

1,132**

1,132

0.24

15

A

Brook trout

Salvelimis fontinalis

1,476**

1,221

0.25

A

Lake trout

Salvelimis namavcush

1,009**

16

G

Neosho mucket

Lampsilis rafinesqueana

1,654**

1,228

0.27

G

Fatmucket

Lampsilis siliquoidea

912.5

17

E

Vernal pool fairy shrimp

Brcmchinecta Ivnchi

1,246

1,246

0.29

18

D

Daphnid

Simocephalus serrulatus

1,291**

1,299

0.31

D

Daphnid

Simocephalus vetulus

1,308**

19

B

Mosquitofish

Gambusia qffinis

1,350

1,350

0.32

20

A

Cutthroat trout

Oncorhvnchus clarkii

1,946**

1,418

0.34

A

Coho salmon

Oncorhvnchus kisutch

1,332**

A

Rainbow trout

Oncorhvnchus mvkiss

1,591

A

Chinook salmon

Oncorhvnchus tshawvtscha

979 9**

21

G

Green floater

Lasmigona subviridis

1,472**

1,472

0.36

22

G

Swamp lymnaea

Lymnaea stagnalis

1,524

1,524

0.37

23

G

Tadpole physa

Phvsella gvrina

1,609

1,609

0.39

24

B

Channel catfish

Ictalurus punctatus

1,617

1,617

0.41

25

G

Pheasantshell

Ortmanniana pectorosa

1,625**

1,625

0.42

26

D

Daphnid

Ceriodaphnia dubia

1,667

1,667

0.44

27

B

Catla

Gibelion catla

1,718**

1,718

0.46

28

C

African clawed frog

Xenopus laevis

1,869**

1,869

0.47

29

E

Amphipod

Gammarus fasciatus

428.1

1,892

0.49

E

Amphipod

Gammarus minus

7,873**

E

Amphipod

Gammarus pseudolimnaeus

2,010**

30

B

Peppered loach

Lepidocephalichthvs
guntea

1,938**

1,938

0.51

31

B

Fathead minnow

Pimephales promelas

1,938

1,938

0.53

32

A

Atlantic salmon

Salmo salar

1,622**

1,969

0.54

A

Brown trout

Salmo trutta

2,390**

33

B

Common carp

Cvprinus carpio

2,075**

2,075

0.56

F-48


-------


MDR









Percentile

Rank

Group1

Name

Species

SMAV

GMAV

Rank

34

B

Goldfish

Carassius auratus

2,152

2,152

0.58

35

D

Daphnid

Daphnia magna

2,183

2,183

0.59

36

B

Walleye

Sander vitreus

2,273**

2,273

0.61

37

C

Indian bullfrog

Euphlyctis hexadactylus

2,331**

2,331

0.63

38

H

Oligochaete

Lumbriculus variegatus

2,368**

2,368

0.64

39

B

White sucker

Catostomus commersonii

2,553**

2,553

0.66

40

B

Black bullhead

Ameiurus melas

2,628**

2,628

0.68

41

B

Flagfish

Jordcmella floridae

2,639**

2,639

0.69

42

B

Medaka

Oryzias latipes

2,670**

2,670

0.71

43

B

Guppy

Poecilia reticulata

2,730**

2,730

0.73

44

B

Zebrafish

Danio rerio

>3,000

>3,000

0.75

45

D

Water flea

Moina macrocopa

3,183**

3,183

0.76

46

B

Rohu

Labeo rohita

3,518**

3,518

0.78

47

B

Mozambique tilapia

Oreochromis mossambicus

1,578**

3,706

0.80

B

Nile tilapia

Oreochromis niloticus

8,705**

48

B

Yellow perch

Percaflavescens

4,050**

4,050

0.81

49

C

Bullfrog

Lithobates catesbeianus

1,791

5,180

0.83

C

Northern leopard frog

Lithobates pipiens

14,976**

50

A

Arctic grayling

Thvmallus arcticus

5,722**

5,722

0.85

51

B

Bluegill

Lepomis macrochirus

6,452

6,452

0.86

52

B

Largemouth bass

Micropterus salmoides

6,684**

6,684

0.88



F

Midge

Paratanvtarsus dissimilis

10,714**





53

F

Midge

Paratanytarsus
parthenogeneticus

5,482

7,664

0.90

54

F

Midge

Chironomus teutons

10,675**

10,675

0.92

55

H

Oligochaete

Limnodrilus hoffmeisteri

13,646**

13,646

0.93

56

E

Amphipod

Crangonvx pseudogracilis

20,152**

20,152

0.95

57

H

Oligochaete

Tubifex tubifex

22,428**

22,428

0.97

58

E

Isopod

Asellus aquaticus

55,185**

55,185

0.98

**Measured EC50 falls outside range of ICE model and a "scaled" EC50 was therefore entered in ICE model.

1: Freshwater MDR Groups

A)	The family Salmonidae in the class Osteichthyes

B)	A second family in the class Osteichthyes, preferably a commercially or recreationally important

warmwater species (e.g., bluegill, channel catfish, etc.)

C)	A third family in the phylum Chordata (may be in the class Osteichthyes or may be an amphibian, etc.)

D)	A planktonic crustacean (e.g., cladoceran, copepod, etc.)

E)	Abenthic crustacean (e.g., ostracod, isopod, amphipod, crayfish, etc.)

F)	An insect (e.g., mayfly, dragonfly, damselfly, stonefly, caddisfly, mosquito, midge, etc.)

G)	A family in a phylum other than Arthropoda or Chordata (e.g., Rotifera, Annelida, Mollusca, etc.)

H)	A family in any order of insect or any phylum not already represented.

F-49


-------
Table F-15. PFBS Final Acute Value and Protective Aquatic Acute Benchmark.

Calculated Freshwater FAY based on 4 lowest values; N=58 GMAVs total





Benchmark calculated by dividing the FAY by 2 and by the sulfonic acid application factor (42.3)





GMAV









Rank

Genus

(mg/L)

ln(GMAV)

ln(GMAV)2

P=R/(N+1)

sqrt(P)

1

Amblema

383.6

5.95

35.40

0.017

0.130

2

Etheostoma

518.0

6.25

39.06

0.034

0.184

3

Margaritifera

706.1

6.56

43.03

0.051

0.225

4

Xyrcnichen

742.7

6.61

43.70

0.068

0.260



£ (Sum):

25.37

161.18

0.17

0.80







P = cumulative probability











R = rank













N = number of GMAVs





S2 =

29.92



S = slope







L =

5.248



L = X-axis intercept





A =

6.471



A = InFAV







FAV =

646.3











FAV/2 =

323.1 mg/L (Preliminary Value)







Adjustment =

323.1 / 42.3 = 7.639 (Preliminary Value / Sulfonic Acid Application Factor)

Benchmark =

7.6 mg/L PFBS (rounded to two significant figures)





F-50


-------
1.00 T
0.90 -j
0.80

« 0.70 --
>

B

3

u

£6
S
«
Pi

-------
Criteria Guidelines (U.S. EPA 1985). The ranked GMAVs for these combined data along with
the MDR met by each GMAV are summarized in Table F-17. GMAVs for the four most
sensitive genera were within a factor of 1.2 of each other (Table F-18). The freshwater FAV (the
5th percentile of the genus sensitivity distribution) for PFHxS is 18.17 mg/L (Table F-18), which
is lower than all of the GMAVs. The FAV was divided by two to obtain a preliminary value of
9.084 mg/L PFHxS and then adjusted by the sulfonic acid application factor (42.3) to obtain the
freshwater acute water column benchmark magnitude of 0.21 mg/L PFHxS (rounded to two
significant figures). This value is expected to be protective of 95% of freshwater genera exposed
to PFHxS under short-term conditions of one-hour duration, if the one-hour average magnitude is
not exceeded more than once in three years (Figure F-6).

F-52


-------
Table F-16. Acceptable models for ICE-estimated Species Sensitivity to PFHxS using the scaling approach for data outside of
the model bounds as in cases indicated.

Bold predicted EC50S used for SMAY calculations.			

Predicted Species

Surrogate

ICE Model Value Range
(mg/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Entered
ECso
(units)

Minimum

Maximum

Bluegill (Lepomis
macrochirus)

Bullfrog (Lithobates
catesbeianus)

1,105**

1,105
(mg/L)

0.003

233

766.4

(378-1,553.6)

Channel catfish (Ictalurus
punctatus)

Bullfrog (Lithobates
catesbeianus)

1,105

1,105,000
(Hg/L)

0.003

20,900

719.8b
(362.4-1,429.7)

Channel catfish (Ictalurus
punctatus)

Zebrafish-embryo (Danio
rerio-embryo)

22.5

22,500
(Ug/L)

0.145

304.5

19.5

(9.3-41)

Daphnid (Daphnia magna)

Bullfrog (Lithobates
catesbeianus)

1,105

1,105,000
(Hg/L)

0.003

20,900

424.1

(68.5-2,627.9)

Daphnid (Daphnia magna)

Zebrafish (Danio rerio)

22.5

22,500
(Ug/L)

0.003

36.91

5.2

(1.2-22.7)

Fairy shrimp

(Streptocephalus

rubricaudatus)

Zebrafish-embryo (Danio
rerio-embryo)

22.5

22,500
(Mg/L)

4.622

5,100.7

114.2

(33.4-390.3)

Fathead minnow (Pimephales
promelas)

Bullfrog (Lithobates
catesbeianus)

1,105

1,105,000
(Hg/L)

0.003

20,900

l,083.6b
(468.5-2,506.4)

Fathead minnow (Pimephales
promelas)

Zebrafish (Danio rerio)

22.5

22,500
(Ug/L)

0.018

25.08

26.0

(11.6-58.2)

Fathead minnow (Pimephales
promelas)

Zebrafish-embryo (Danio
rerio-embryo)

22.5

22,500
(Hg/L)

0.023

54,579

18.3

(13.3-25.2)

Flagfish (Jordanella floridae)

Zebrafish (Danio rerio)

22.5

22,500
(Ug/L)

7.797

649.3

33.9

(19.7-58.4)

Goldfish (Carassius auratus)

Bullfrog (Lithobates
catesbeianus)

1,105

1,105,000
(Hg/L)

0.003

20,900

l,027.7b
(405.9-2,602.1)

Goldfish (Carassius auratus)

Zebrafish-embryo (Danio
rerio-embryo)

22.5

22,500
(Ug/L)

0.145

304.5

37.3

(23.2-59.9)

Midge (Paratanytarsus
dissimilis)

Bullfrog (Lithobates
catesbeianus)

1,105

1,105,000
(Hg/L)

0.003

3,020.0

1,069.7

(176.6-6,480.9)

F-53


-------
Predicted Species

Surrogate

ICE Model Value Range
(mg/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Entered
ECso
(units)

Minimum

Maximum

Mosquitofish (Gambusia
affinis)

Bullfrog (Lithobates
catesbeianus)

1,105

1,105,000
(Hg/L)

18.6

20,900

805.1

(394.5-1,643.4)

Oligochaete (Limnodrilus
hoffmeisteri)

Zebrafish-embryo (Danio
rerio-embryo)

22.5

22,500
(ug/L)

0.145

239.3

63.2

(9.9-403.1)

Rainbow trout
(Oncorhynchus mykiss)

Bullfrog (Lithobates
catesbeianus)

1,105

1,105,000
(Hg/L)

0.003

20,900

713.5b
(200.3-2,541.4)

Rainbow trout
(Oncorhynchus mykiss)

Zebrafish (Danio rerio)

22.5

22,500
(UR/L)

0.003

26.39

38.3

(9.5-155.1)

Rainbow trout
(Oncorhynchus mykiss)

Zebrafish-embryo (Danio
rerio-embryo)

22.5

22,500
(Hg/L)

0.023

8,843.9

9.6

(4.2-22)

**Measured ECso falls outside range of ICE model and a "scaled" ECso was therefore entered in ICE model.
a Saltwater surrogate species were not used.

b Not used because other more closely taxonomically-related models were available.

0 Not used because other more closely taxonomically-related models where the measured surrogate ECso values fell within the ICE model range were available.

F-54


-------
Table F-17. Ranked PFHxS Genus Mean Acute Values.

Values in bold and highlighted are derived from empirical PFHxS toxicity tests with the species.

Rank

MDR

Group1

Name

Species

SMAV

GMAV

Percentile
Rank

1

A

Rainbow trout

Oncorhvnchus mvkiss

19.19

19.19

0.07

2

B

Channel catfish

Ictalurus punctatus

19.46

19.46

0.14

3

B

Fathead minnow

Pimephales promelas

21.80

21.80

0.21

4

B

Zebrafish

Danio rerio

22.50

22.50

0.29

5

B

Flagfish

Jordcmella floridae

33.90

33.90

0.36

6

B

Goldfish

Carassius auratus

37.28

37.28

0.43

7

D

Daphnid

Daphnia magna

46.87

46.87

0.50

8

H

Oligochaete

Limnodrilus hoffmeisteri

63.23

63.23

0.57

9

E

Fairy shrimp

Streptocephalus
proboscideus

114.2

114.2

0.64

10

B

Bluegill

Lepomis macrochirus

766.4**

766.4

0.71

11

B

Mosquitofish

Gambusia affmis

805.1

805.1

0.79

12

C

Bullfrog

Lithobates catesbeiana

1,105

915.2

0.86

C

Green frog

Lithobates clamitans

758.0

13

F

Midge

Paratanvtarsus dissimilis

1,070

1,070

0.93

**Measured EC50 falls outside range of ICE model and a "scaled" EC50 was therefore entered in ICE model.
1: Freshwater MDR Groups

A)	The family Salmonidae in the class Osteichthyes

B)	A second family in the class Osteichthyes, preferably a commercially or recreationally important

warmwater species (e.g., bluegill, channel catfish, etc.)

C)	A third family in the phylum Chordata (may be in the class Osteichthyes or may be an amphibian, etc.)

D)	A planktonic crustacean (e.g., cladoceran, copepod, etc.)

E)	Abenthic crustacean (e.g., ostracod, isopod, amphipod, crayfish, etc.)

F)	An insect (e.g., mayfly, dragonfly, damselfly, stonefly, caddisfly, mosquito, midge, etc.)

G)	A family in a phylum other than Arthropoda or Chordata (e.g., Rotifera, Annelida, Mollusca, etc.)

H)	A family in any order of insect or any phylum not already represented.

F-55


-------
Table F-18. PFHxS Final Acute Value and Protective Aquatic Acute Benchmark.

Bold values represent genera for which empirical toxicity data were available.

Note: Missing MDR Group: G	

Calculated Freshwater FAY based on 4 lowest values; N=13 GMAVs total

Benchmark calculated by dividing the FAY by 2 and by the sulfonic acid application factor (42.3)





GMAV









Rank

Genus

(mg/L)

ln(GMAV)

ln(GMAV)2

P=R/(N+1)

sqrt(P)

1

Oncorhynchns

19.19

2.95

8.73

0.071

0.267

2

Ictalurus

19.46

2.97

8.81

0.143

0.378

3

Pimephales

21.80

3.08

9.50

0.214

0.463

4

Danio

22.50

3.11

9.69

0.286

0.535



£ (Sum):

12.12

36.73

0.71

1.64

S2 =
L =
A =
FAV =
FAV/2 =
Adjustment =
Benchmark =

P = cumulative probability
R = rank

N = number of GMAVs
0.48	S = slope

2.744	L = X-axis intercept

2.900	A = InFAV

18.17

9.084 mg/L (Preliminary Value)

9.084 / 42.3 = 0.2147 (Preliminary Value / Sulfonic Acid Application Factor)
0.21 mg/L PFHxS (rounded to two significant figures)	

F-56


-------
s

3

u

-£

a

C3
<&

-------
The combined empirical and ICE data resulted in 55 GMAVs that collectively fulfill the
eight MDR groups for deriving a freshwater criterion as defined by the Aquatic Life Criteria
Guidelines (U.S. EPA 1985). The ranked GMAVs for these combined data along with the MDR
met by each GMAV is summarized in Table F-20. GMAVs for the four most sensitive genera
were within a factor of 1.8 of each other (Table F-21). The freshwater FAV (the 5th percentile of
the genus sensitivity distribution) for 8:2 FTUCA is 1.292 mg/L (Table F-21), which is lower
than all of the GMAVs except for the ICE-derived species value for the threeridge mussel,
Amblemaplicata, GMAV = 0.79 mg/L. The FAV was divided by two to obtain a preliminary
value of 0.6458 mg/L 8:2 FTUCA and then adjusted by the carboxylic acid application factor
(15.8) to obtain the freshwater acute water column benchmark magnitude of 0.041 mg/L 8:2
FTUCA (rounded to two significant figures). This value is expected to be protective of 95% of
freshwater genera exposed to 8:2 FTUCA under short-term conditions of one-hour duration, if
the one-hour average magnitude is not exceeded more than once in three years (Figure F-7).

F-58


-------
Table F-19. Acceptable models for ICE-estimated Species Sensitivity to 8:2 FTUCA using the scaling approach for data
outside of the model bounds as in cases indicated.

Bold predicted EC50S used for SMAY calculations.			

Predicted Species

Surrogate

ICE Model Value Range
(mg/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Entered
ECso
(units)

Minimum

Maximum

Amphipod (Crangonyx
pseudogracilis)

Daphnid (Daphnia magna)

3.2

3,200
(ug/L)

0.0008

166.8

28.6

(7-116.1)

Amphipod (Gammarus
fasciatus)

Daphnid (Daphnia magna)

3.2

3,200
(Hg/L)

0.0002

5,000

2.0

(1-4.1)

Amphipod (Gammarus
pseudolimnaeus)

Daphnid (Daphnia magna)

3.2

3,200
(ug/L)

0.0001

68.3

2.9

(1-7.8)

Apache trout (Oncorhynchus
gilae)*

Rainbow trout
(Oncorhynchus mykiss)

81**

81
(mg/L)

0.004

1.625

55.5
(35-87.9)

Atlantic salmon (Salmo salar)

Rainbow trout
(Oncorhynchus mykiss)

81

81,000
(ug/L)

0.0002

95.86

79.3

(36.6-171.6)

Beaver-tail fairy shrimp
(Thamnocephalus platyurus)

Daphnid (Daphnia magna)

3.2

3,200
(n.g/L)

0.0003

8,694.5

2.6

(1.8-3.9)

Black bullhead (Ameiurus
melas)

Rainbow trout
(Oncorhynchus mykiss)

81**

81
(mg/L)

<0.0001

9.81

411.3

(105.2-1,607.8)

Bluegill (Lepomis
macrochirus)

Daphnid (Daphnia magna)

3.2

3,200
(n.g/L)

0.0001

46,278

4.9b
(3.7-6.4)

Bluegill (Lepomis
macrochirus)

Rainbow trout
(Oncorhynchus mykiss)

81

81,000
(ug/L)

0.0001

8,341.5

79.0

(64.3-97.1)

Bonytail (Gila elegans)

Rainbow trout
(Oncorhynchus mykiss)

81**

81
(mg/L)

0.003

43.07

288.8

(102.7-811.8)

Brook trout (Salvelinus
fontinalis)

Rainbow trout
(Oncorhynchus mykiss)

81

81,000
(n.g/L)

0.0006

96.69

79.4

(44.1-142.9)

Brown trout (Salmo trutta)

Rainbow trout
(Oncorhynchus mykiss)

81

81,000
(n.g/L)

0.0002

95.86

83.8

(47-149.4)

Bullfrog (Lithobates
catesbeianus)

Rainbow trout
(Oncorhynchus mykiss)

81

81,000
(n.g/L)

0.0006

13,400

132.5

(51-344.2)

F-59


-------
Predicted Species

Surrogate

ICE Model Value Range
(mg/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Entered
ECso
(units)

Minimum

Maximum

Cape Fear shiner (Notropis
mekistocholas)

Rainbow trout
(Oncorhynchus mykiss)

81**

81
(mg/L)

0.004

1.625

102.3

(69.6-150.5)

Channel catfish (Ictalurus
punctatus)

Rainbow trout
(Oncorhynchus mykiss)

81

81,000
(ug/L)

<0.0001

13,400

67.4

(39.8-114)

Chinook salmon

(Oncorhynchus

tshawytscha)*

Rainbow trout
(Oncorhynchus mykiss)

81

81,000
(Mg/L)

0.003

724.4

102.9
(57.1-185.4)

Coho salmon (Oncorhynchus
kisutch)*

Rainbow trout
(Oncorhynchus mykiss)

81**

81
(mg/L)

<0.0001

43.07

106.1
(91.6-122.9)

Colorado squawfish
(Ptychocheilus lucius)

Rainbow trout
(Oncorhynchus mykiss)

81**

81
(mg/L)

0.003

43.07

453.5

(101.9-2,018.3)

Common carp (Cyprinus
carpio)

Rainbow trout
(Oncorhynchus mykiss)

81

81,000
(Hg/L)

0.0002

198.4

75.2

(31-182.2)

Cutthroat trout
(Oncorhynchus clarkii)*

Rainbow trout
(Oncorhynchus mykiss)

81

81,000
(Hg/L)

<0.0001

198.4

65.5
(42.9-100)

Cuvier's foam froglet
(Physalaemus cuvieri)

Rainbow trout
(Oncorhynchus mykiss)

81

81,000
(Hg/L)

0.7099

155.5

91.1

(48.4-171.5)

Daphnid (Ceriodaphnia
dubia)

Daphnid (Daphnia magna)

3.2

3,200
(Hg/L)

0.0003

46,278

2.1

(1.5-2.9)

Daphnid (Ceriodaphnia
reticulata)

Daphnid (Daphnia magna)

3 2**

3.2
(mg/L)

0.0008

0.232

2.6

(0.6-11.3)

Daphnid (Daphnia galeata)*

Daphnid (Daphnia magna)

3 2**

3.2
(mg/L)

0.0001

0.646

5.4
(2.1-14.2)

Daphnid (Daphnia
longispina)*

Daphnid (Daphnia magna)

3.2

3,200
(Hg/L)

0.009

10.36

4.9
(1-23.7)

Daphnid (Daphnia pulex)*

Daphnid (Daphnia magna)

3.2

3,200
(Hg/L)

0.0002

4,894.7

2.4
(1.7-3.3)

Daphnid (Daphnia pulicaria)*

Daphnid (Daphnia magna)

3.2

3,200
(Hg/L)

0.014

281.6

2.6
(0.7-9.5)

Daphnid (Simocephalus
serrulatus)

Daphnid (Daphnia magna)

3.2

3,200
(Hg/L)

0.0002

7.2

1.9

(0.7-5.2)

F-60


-------
Predicted Species

Surrogate

ICE Model Value Range
(mg/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Entered
ECso
(units)

Minimum

Maximum

Daphnid (Simocephalus
vetulus)

Daphnid (Daphnia magna)

3.2

3,200
(Hg/L)

0.0001

166.8

1.7

(0.3-12.3)

Fathead minnow (Pimephales
promelas)

Daphnid (Daphnia magna)

3.2

3,200
(ug/L)

0.0002

46,500

7.0b
(5.4-9)

Fathead minnow (Pimephales
promelas)

Rainbow trout
(Oncorhynchus mykiss)

81

81,000
(Hg/L)

0.0002

13,400

107.7

(79.8-145.3)

Fatmucket (Lampsilis
siliquoidea)

Daphnid (Daphnia magna)

3.2

3,200
(ug/L)

0.014

8,694.5

2.5

(1.6-4.1)

Goldfish (Carassius auratus)

Rainbow trout
(Oncorhynchus mykiss)

81

81,000
(n.g/L)

<0.0001

13,400

132.7

(62.2-283.3)

Green floater (Lasmigona
subviridis)

Daphnid (Daphnia magna)

3.2

3,200
(ug/L)

0.014

166.8

1.9

(0.4-9.4)

Green sunfish (Lepomis
cyanellus)

Rainbow trout
(Oncorhynchus mykiss)

81**

81
(mg/L)

<0.0001

19.93

208

(129.4-334.4)

Greenthroat darter
(Etheostoma lepidum)

Rainbow trout
(Oncorhynchus mykiss)

81**

81
(mg/L)

0.004

1.625

92.6

(31.2-274.7)

Guppy (Poecilia reticulata)

Rainbow trout
(Oncorhynchus mykiss)

81

81,000
(n.g/L)

0.0006

198.4

54.9

(12.3-243.7)

Indian bullfrog (Euphlyctis
hexadactylus)

Rainbow trout
(Oncorhynchus mykiss)

81**

81
(mg/L)

0.028

19.93

184.0

(26-1,301.7)

Isopod (Asellus aquaticus)

Daphnid (Daphnia magna)

3.2

3,200
(n.g/L)

0.0003

166.8

76.6

(23-255.1)

Isopod (Caecidotea
brevicauda)

Rainbow trout
(Oncorhynchus mykiss)

81

81,000
(ug/L)

0.0006

82

16.8

(3.2-89.7)

Lake trout (Salvelinus
namaycush)

Rainbow trout
(Oncorhynchus mykiss)

81

81,000
(n.g/L)

0.0002

198.4

41.3

(24.5-69.8)

Largemouth bass
(Micropterus salmoides)

Rainbow trout
(Oncorhynchus mykiss)

81

81,000
(ug/L)

<0.0001

95.86

46.8

(20.7-106.2)

Mayfly (Drunella grandis)

Rainbow trout
(Oncorhynchus mykiss)

81A

81
(mg/L)

0.0006

95.86

365.0

(106.1-1,255.8)

F-61


-------
Predicted Species

Surrogate

ICE Model Value Range
(mg/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Entered
ECso
(units)

Minimum

Maximum

Midge (Paratanytarsus
dissimilis)

Rainbow trout
(Oncorhynchus mykiss)

81

81,000
(Hg/L)

0.0006

1,330

241.0

(72.2-804.7)

Midge (Paratanytarsus
parthenogeneticus)

Daphnid (Daphnia magna)

3.2

3,200
(ug/L)

0.37

14,500

9.3

(3.5-24.7)

Mississippi grass shrimp
(Palaemonetes kadiakensis)

Daphnid (Daphnia magna)

3.2

3,200
(Hg/L)

0.0003

58

1.3

(0.3-6.2)

Mrigal carp (Cirrhinus
mrigala)

Rainbow trout
(Oncorhynchus mykiss)

81**

81
(mg/L)

0.0006

1.625

298.8

(53.6-1,666.3)

Neosho mucket (Lampsilis
rafinesqueana)

Daphnid (Daphnia magna)

3.2

3,200
(Hg/L)

0.042

166.8

2.3

(1.1-5.1)

Northern leopard frog
(Lithobates pipiens)

Rainbow trout
(Oncorhynchus mykiss)

81

81,000
(Ug/L)

0.002

198.4

125.5

(62.3-252.8)

Northern pike (Esox lucius)

Rainbow trout
(Oncorhynchus mykiss)

81**

81
(mg/L)

0.005

1.822

48.1

(16.4-141.3)

Oligochaete (Limnodrilus
hoffmeisteri)

Daphnid (Daphnia magna)

3.2

3,200
(Ug/L)

0.003

281.6

17.9

(6.9-46.2)

Oligochaete (Limnodrilus
hoffmeisteri)

Rainbow trout
(Oncorhynchus mykiss)

81A

81
(mg/L)

0.003

707

889.5b
(128.1-6,176.8)

Oligochaete (Tubifex tubifex)

Daphnid (Daphnia magna)

3.2

3,200
(Ug/L)

0.0001

4,894.7

31.6

(7.4-134.7)

Oriental river shrimp
(Macrobrachium nipponense)

Daphnid (Daphnia magna)

3.2

3,200
(Hg/L)

0.011

281.6

1.5

(0.7-3.4)

Paper pondshell (Utterbackia
imbecillis)

Daphnid (Daphnia magna)

3.2

3,200
(Ug/L)

0.014

8,694.5

2.5

(1.6-3.7)

Pheasantshell (Ortmanniana
pectorosa)

Daphnid (Daphnia magna)

3.2

3,200
(Hg/L)

0.042

545.9

2.4

(0.8-7)

Rainbow trout
(Oncorhynchus mykiss)*

Daphnid (Daphnia magna)

3 2**

3200
( Ug/L)

0.0001

14,500

3.4
(2.7-4.3)

Razorback sucker
(Xyrauchen texanus)

Rainbow trout
(Oncorhynchus mykiss)

81**

81
(mg/L)

0.003

43.07

250.4

(96.5-649.5)

F-62


-------
Predicted Species

Surrogate

ICE Model Value Range
(mg/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Entered
ECso
(units)

Minimum

Maximum

Riceland prawn
(Macrobrachium lanchesteri)

Rainbow trout
(Oncorhynchus mykiss)

81**

81
(mg/L)

0.003

2.04

392.0

(87.1-1,764.3)

Rohu (Labeo rohita)

Rainbow trout
(Oncorhynchus mykiss)

81**

81
(mg/L)

0.0006

1.625

818.9

(162.1-4,136.2)

Shortnose sturgeon
(Acipenser brevirostrum)

Rainbow trout
(Oncorhynchus mykiss)

81

81,000
(Hg/L)

0.028

95.86

134.5

(26.7-677.1)

Silver perch (Bidyanus
bidyanus)

Rainbow trout
(Oncorhynchus mykiss)

81**

81
(mg/L)

0.001

7.075

110.5

(70.9-172.1)

Sockeye salmon
(Oncorhynchus nerka)*

Rainbow trout
(Oncorhynchus mykiss)

81**

81
(mg/L)

0.028

7.5

163.6
(50.5-529.6)

Southern leopard frog
(Lithobates sphenocephalus)

Rainbow trout
(Oncorhynchus mykiss)

81**

81
(mg/L)

0.028

9.7

151.5

(76.9-298.6)

Spotfin chub (Erimonax
monachus)

Rainbow trout
(Oncorhynchus mykiss)

81**

81
(mg/L)

0.004

1.625

74.3

(33.2-166.2)

Swamp lymnaea (Lymnaea
stagnalis)

Daphnid (Daphnia magna)

3.2

3,200
(Ug/L)

0.014

8,694.5

3.2

(1.2-8.4)

Tadpole physa (Physella
gyrina)

Daphnid (Daphnia magna)

3.2

3,200
(Hg/L)

0.014

8,694.5

3.0

(1.4-6.7)

Threeridge (Amblema
plicata)

Daphnid (Daphnia magna)

3.2

3,200
(Ug/L)

0.014

4,894.7

0.8

(0.3-2.1)

Vernal pool fairy shrimp
(Branchinecta lynchi)

Daphnid (Daphnia magna)

3.2

3,200
(Hg/L)

0.014

8,694.5

2.6

(1.4-4.9)

Walleye (Sander vitreus)

Rainbow trout
(Oncorhynchus mykiss)

81*

81
(mg/L)

0.005

16.24

78.4

(27.6-222.5)

Washboard (Megalonaias
nervosa)

Daphnid (Daphnia magna)

3.2

3,200
(Hg/L)

0.014

8,694.5

1.4

(0.8-2.6)

Water flea (Chydorus
sphaericus)

Daphnid (Daphnia magna)

3.2

3,200
(Ug/L)

0.009

977.6

1.3

(0.8-2.2)

Water flea (Moina
macrocopa)

Daphnid (Daphnia magna)

3.2

3,200
(Hg/L)

0.009

281.6

4.2

(1.3-13.2)

F-63


-------
Predicted Species

Surrogate

ICE Model Value Range
(mg/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Entered
ECso
(units)

Minimum

Maximum

Western pearlshell
(Margaritifera falcata)

Daphnid (Daphnia magna)

3.2

3,200
(Hg/L)

0.014

8,694.5

2.3

(1.1-4.6)

Western toad (Anaxyrus
boreas)

Rainbow trout
(Oncorhynchus mykiss)

81**

81
(mg/L)

0.028

1.625

107.9

(25.9-450.6)

Yellow perch (Perca
flavescens)

Rainbow trout
(Oncorhynchus mykiss)

81**

81
(mg/L)

<0.0001

16.24

102.6

(49.6-212)

Zebrafish (Danio rerio)

Daphnid (Daphnia magna)

3.2

3,200
(ug/L)

0.0001

50

8.3b
(2.5-28)

Zebrafish (Danio rerio)

Rainbow trout
(Oncorhynchus mykiss)

81

81,000
(Hg/L)

0.0002

229.1

34.6

(11.3-106)

Zebrafish-embryo (Danio
rerio-embryo)

Daphnid (Daphnia magna)

3.2

3,200
(ug/L)

0.0001

46,500

10.7b
(5.7-19.9)

Zebrafish-embryo (Danio
rerio-embryo)

Rainbow trout
(Oncorhynchus mykiss)

81

81,000
(Hg/L)

0.0002

8,341.5

65.2

(29-146.3)

* Acceptable models that were not used because genus level empirical data were available.

**Measured EC50 falls outside range of ICE model and a "scaled" ECso was therefore entered in ICE model.

A |ig/L CI greater than 50-fold. Used mg/L with acceptable CI.
a Saltwater surrogate species were not used.

b Not used because other more closely taxonomically-related models were available.

0 Not used because other more closely taxonomically-related models where the measured surrogate ECso values fell within the ICE model range were available.

F-64


-------
Table F-20. Ranked 8:2 FTUCA Genus Mean Acute Values.

Values in bold and highlighted are derived from empirical 8:2 FTUCA

1 the species

Rank

MDR

Group1

Name

Species

SMAV

GMAV

Percentile
Rank

1

G

Threeridge

Amblema plicata

0.7900

0.7900

0.02

2

E

Mississippi grass shrimp

Palaemonetes kadiakensis

1.330

1.330

0.04

3

D

Daphnid

Chydorus sphaericus

1.340

1.340

0.05

4

G

Washboard

Megalonaias nen'osa

1.400

1.400

0.07

5

D

Daphnid

Simocephalus serrulatus

1.850

1.794

0.09

D

Daphnid

Simocephalus vetulus

1.740

6

G

Green floater

Lasmigona subviridis

1.900

1.900

0.11

7

G

Western pearlshell

Margaritifera falcata

2.270

2.270

0.13

8

D

Daphnid

Ceriodaphnia reticulata

2.560**

2.302

0.14

D

Daphnid

Ceriodaphnia dubia

2.070

9

G

Pheasantshell

Ortmanniana pectorosa

2.350

2.350

0.16

10

E

Amphipod

Gammarus fasciatus

2.040

2.411

0.18

E

Amphipod

Gammarus pseudolimnaeus

2.850

11

G

Neosho mucket

Lampsilis rafinesqueana

2.320

2.428

0.20

G

Fatmucket

Lampsilis siliquoidea

2.540

12

G

Paper pondshell

Utterbackia imbecillis

2.450

2.450

0.21

13

E

Beaver-tail fairy shrimp

Thamnocephalus platvurus

2.630

2.630

0.23

14

E

Vernal pool fairy shrimp

Branchinecta Ivnchi

2.640

2.640

0.25

15

G

Tadpole physa

Phvsella gvrina

3.030

3.030

0.27

16

D

Daphnid

Daphnia magna

3.200

3.200

0.29

17

G

Swamp lymnaea

Lymnaea stagnalis

3.210

3.210

0.30

18

D

Water flea

Moina macrocopa

4.190

4.190

0.32

19

E

Isopod

Caecidotea brevicauda

16.82

16.82

0.34

20

H

Oligochaete

Limnodrilus hoffmeisteri

17.89

17.89

0.36

21

E

Riceland prawn

Macrobrachium
lanchesteri

392.0**

24.41

0.375

E

Oriental river shrimp

Macrobrachium
nipponense

1.520

22

E

Amphipod

Crangonyx pseudogracilis

28.57

28.57

0.39

23

H

Oligochaete

Tubifex tubifex

31.57

31.57

0.41

24

B

Largemouth bass

Micropterus salmoides

46.83

46.83

0.43

25

F

Midge

Paratanvtarsus dissimilis

241.0

47.44

0.45

F

Midge

Paratanytarsus
parthenogeneticus

9.340

26

B

Zebrafish

Danio rerio

47.48

47.48

0.46

27

B

Northern pike

Esox lucius

48.13**

48.13

0.48

28

B

Guppy

Poecilia reticulata

54.85

54.85

0.50

29

A

Brook trout

Salvelinus fontinalis

79.36

57.25

0.52

A

Lake trout

Salvelinus namavcush

41.30

30

B

Channel catfish

Ictalurus punctatus

67.38

67.38

0.54

31

B

Spotfin chub

Erimonax monachus

74.30**

74.30

0.55

32

B

Common carp

Cyprinus carpio

75.18

75.18

0.57

33

E

Isopod

Asellus aquaticus

76.56

76.56

0.59

34

B

Walleye

Sander vitreus

78.36**

78.36

0.61

F-65


-------
Rank

MDR

Group1

Name

Species

SMAV

GMAV

Percentile
Rank

35

A

Rainbow trout

Oncorhynchus mykiss

81.00

81.00

0.63

36

A

Atlantic salmon

Salmo salar

79.29

81.49

0.64

A

Brown trout

Salmo trutta

83.76

37

C

Cuvier's foam froglet

Phvsalaemus cuvieri

91.14

91.14

0.66

38

B

Greenthroat darter

Etheostoma lepidum

92.55**

92.55

0.68

39

B

Cape Fear shiner

Notropis mekistocholas

102.3**

102.3

0.70

40

B

Yellow perch

Percaflavescens

102.6**

102.6

0.71

41

B

Fathead minnow

Pimephales promelas

107.7

107.7

0.73

42

C

Western toad

Anaxvrus boreas

107.9**

107.9

0.75

43

B

Silver perch

Bidvamis bidvanus

110.5**

110.5

0.77

44

B

Green sunfish

Lepomis cyanellus

208.0**

128.2

0.79

B

Bluegill

Lepomis macrochirus

78.99

45

B

Goldfish

Carassius auratus

132.7

132.7

0.80

46

B

Shortnose sturgeon

Acipenser brevirostrum

134.5

134.5

0.82

47

C

Bullfrog

Lithobates catesbeianus

132.5

136.0

0.84

C

Northern leopard frog

Lithobates pipiens

125.5

C

Southern leopard frog

Lithobates sphenocephalus

151.5**

48

C

Indian bullfrog

Euphlyctis hexadactvlus

184.0**

184.0

0.86

49

B

Razorback sucker

Xwauchen texanus

250.4**

250.4

0.88

50

B

Bonytail

Gila elegans

288.8**

288.8

0.89

51

B

Mrigal carp

Cirrhinus mrigala

298.8**

298.8

0.91

52

F

Mayfly

Drunella grandis

365.0

365.0

0.93

53

B

Black bullhead

Ameiurus melas

411.3**

411.3

0.95

54

B

Colorado squawfish

Ptvchocheilus lucius

453.5**

453.5

0.96

55

B

Rohu

Labeo rohita

818.9**

818.9

0.98

**Measured EC50 falls outside range of ICE model and a "scaled" EC50 was therefore entered in ICE model.

1: Freshwater MDR Groups

A)	The family Salmonidae in the class Osteichthyes

B)	A second family in the class Osteichthyes, preferably a commercially or recreationally important

warmwater species (e.g., bluegill, channel catfish, etc.)

C)	A third family in the phylum Chordata (may be in the class Osteichthyes or may be an amphibian, etc.)

D)	A planktonic crustacean (e.g., cladoceran, copepod, etc.)

E)	Abenthic crustacean (e.g., ostracod, isopod, amphipod, crayfish, etc.)

F)	An insect (e.g., mayfly, dragonfly, damselfly, stonefly, caddisfly, mosquito, midge, etc.)

G)	A family in a phylum other than Arthropoda or Chordata (e.g., Rotifera, Annelida, Mollusca, etc.)

H)	A family in any order of insect or any phylum not already represented.

F-66


-------
Table F-21. 8:2 FTUCA Final Acute Value and Protective Aquatic Acute Benchmark.

Calculated Freshwater FAY based on 4 lowest values; N=55 GMAVs total





Benchmark calculated by dividing the FAY by 2 and by the carboxylic acid application factor (15.8)





GMAV









Rank

Genus

(mg/L)

ln(GMAV)

ln(GMAV)2

P=R/(N+1)

sqrt(P)

1

Amblema

0.79

-0.24

0.06

0.018

0.134

2

Palaemonetes

1.33

0.29

0.08

0.036

0.189

3

Chydorus

1.34

0.29

0.09

0.054

0.231

4

Megalonaias

1.40

0.34

0.11

0.071

0.267



£ (Sum):

0.68

0.34

0.18

0.82







P = cumulative probability











R = rank













N = number of GMAVs





S2 =

22.23



S = slope







L =

-0.798



L = X-axis intercept





A =

0.256



A = InFAV







FAV =

1.292











FAV/2 =

0.6458 mg/L (Preliminary Value)







Adjustment =

0.6458 / 15.8 = 0.04087 (Preliminary Value / Carboxylic Acid Application Factor)

Benchmark =

0.041 mg/L 8:2 FTUCA (rounded to two significant figures)



F-67


-------
1.00 -r

0.90

0.80

« 0.70 -
>

s

3

u

s

rt

06


-------
The combined empirical and ICE data resulted in 59 GMAVs that collectively fulfill the
eight MDR groups for deriving a freshwater criterion as defined by the Aquatic Life Criteria
Guidelines (U.S. EPA 1985). The ranked GMAVs for these combined data along with the MDR
met by each GMAV are summarized in Table F-23. GMAVs for the four most sensitive genera
were within a factor of 2.1 of each other (Table F-24). The freshwater FAV (the 5th percentile of
the genus sensitivity distribution) for 7:3 FTCA is 0.4609 mg/L (Table F-24), which is lower
than all of the GMAVs except for three of the ICE-derived species values (the threeridge mussel,
Amblemaplicata, GMAV=0.25 mg/L, the water flea Chydorus sphaericus, GMAV=0.43 mg/L,
and the washboard, Megalonaias nervosa, GMAV=0.44 mg/L). The FAV was divided by two to
obtain a preliminary value of 0.2305 mg/L 7:3 FTCA and then adjusted by the carboxylic acid
application factor (15.8) to obtain the freshwater acute water column benchmark magnitude of
0.015 mg/L 7:3 FTCA (rounded to two significant figures). This value is expected to be
protective of 95% of freshwater genera exposed to 7:3 FTCA under short-term conditions of one-
hour duration, if the one-hour average magnitude is not exceeded more than once in three years
(Figure F-8).

F-69


-------
Table F-22. Acceptable models for ICE-estimated Species Sensitivity to 7:3 FTCA using the scaling approach for data outside
of the model bounds as in cases indicated.

Bold predicted EC50S used for SMAY calculations.			

Predicted Species

Surrogate

ICE Model Value Range
(mg/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Entered
ECso
(units)

Minimum

Maximum

Amphipod (Crangonyx
pseudogracilis)

Daphnid (Daphnia magna)

0.9592

959.2
(ug/L)

0.0008

166.8

9.5

(2.8-32.4)

Amphipod (Gammarus
fasciatus)

Daphnid (Daphnia magna)

0.9592

959.2
(Hg/L)

0.0002

5,000

0.8

(0.4-1.4)

Amphipod (Gammarus
pseudolimnaeus)

Daphnid (Daphnia magna)

0.9592

959.2
(ug/L)

0.0001

68.3

1.0

(0.4-2.2)

Apache trout (Oncorhynchus
gilae)*

Rainbow trout
(Oncorhynchus mykiss)

32**

32
(mg/L)

0.0041

1.625

20.0
(12.3-32.6)

Atlantic salmon (Salmo salar)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(ug/L)

0.0002

95.86

31.0

(15.7-61.3)

Beaver-tail fairy shrimp
(Thamnocephalus platyurus)

Daphnid (Daphnia magna)

0.9592

959.2
(n.g/L)

0.0003

8,694.5

0.9

(0.6-1.3)

Black bullhead (Ameiurus
melas)

Rainbow trout
(Oncorhynchus mykiss)

32**

32
(mg/L)

<0.0001

9.81

207.8

(56.1-770)

Bluegill (Lepomis
macrochirus)

Daphnid (Daphnia magna)

0.9592

959.2
(n.g/L)

0.0001

46,278

2.1b
(1.6-2.7)

Bluegill (Lepomis
macrochirus)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(ug/L)

0.0001

8,341.5

33.1

(27.6-39.6)

Bonytail (Gila elegans)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(n.g/L)

0.003

43.07

72.7

(12.6-420.2)

Brook trout (Salvelinus
fontinalis)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(n.g/L)

0.0006

96.69

31.4

(18.8-52.5)

Brown trout (Salmo trutta)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(n.g/L)

0.0002

95.86

33.1

(19.8-55.3)

Bullfrog (Lithobates
catesbeianus)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(n.g/L)

0.0006

13,400

58.8

(23.7-145.4)

F-70


-------
Predicted Species

Surrogate

ICE Model Value Range
(mg/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Entered
ECso
(units)

Minimum

Maximum

Cape Fear shiner (Notropis
mekistocholas)

Rainbow trout
(Oncorhynchus mykiss)

32**

32
(mg/L)

0.004

1.625

34.4

(22.8-51.9)

Channel catfish (Ictalurus
punctatus)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(ug/L)

<0.0001

13,400

31.6

(19.9-50.1)

Chinook salmon

(Oncorhynchus

tshawytscha)*

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(Mg/L)

0.003

724.4

40.0
(24-66.9)

Coho salmon (Oncorhynchus
kisutch)*

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(Hg/L)

<0.0001

43.07

49.4
(38.4-63.6)

Colorado squawfish
(Ptychocheilus lucius)

Rainbow trout
(Oncorhynchus mykiss)

32A

32
(mg/L)

0.003

43.07

195.1

(34.5-1,103.6)

Common carp (Cyprinus
carpio)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(Hg/L)

0.0002

198.4

35.6

(16.3-77.7)

Cutthroat trout
(Oncorhynchus clarkii)*

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(Hg/L)

<0.0001

198.4

27.1
(18.7-39.3)

Cuvier's foam froglet
(Physalaemus cuvieri)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(Hg/L)

0.7099

155.5

51.8

(30.8-87.1)

Daphnid (Ceriodaphnia
dubia)

Daphnid (Daphnia magna)

0.9592

959.2
(Hg/L)

0.0003

46,278

0.6

(0.4-0.9)

Daphnid (Daphnia galeata)*

Daphnid (Daphnia magna)

0.9592**

0.9592
(mg/L)

0.0001

0.646

1.8
(0.6-5.8)

Daphnid (Daphnia
longispina)*

Daphnid (Daphnia magna)

0.9592

959.2
(Hg/L)

0.009

10.36

1.1
(0.3-4.3)

Daphnid (Daphnia pulex)*

Daphnid (Daphnia magna)

0.9592

959.2
(Hg/L)

0.0002

4,894.7

0.7
(0.5-1)

Daphnid (Daphnia pulicaria)*

Daphnid (Daphnia magna)

0.9592

959.2
(Hg/L)

0.014

281.6

0.7
(0.2-3.1)

Daphnid (Simocephalus
serrulatus)

Daphnid (Daphnia magna)

0.9592

959.2
(Hg/L)

0.0002

7.2

0.6

(0.3-1.4)

Daphnid (Simocephalus
vetulus)

Daphnid (Daphnia magna)

0.9592

959.2
(Hg/L)

0.0001

166.8

0.7

(0.1-3.8)

F-71


-------
Predicted Species

Surrogate

ICE Model Value Range
(mg/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Entered
ECso
(units)

Minimum

Maximum

Fathead minnow (Pimephales
promelas)

Daphnid (Daphnia magna)

0.9592

959.2
(Hg/L)

0.0002

46,500

3.0b
(2.3-3.9)

Fathead minnow (Pimephales
promelas)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(Ug/L)

0.0002

13,400

46.7

(36-60.7)

Fatmucket (Lampsilis
siliquoidea)

Daphnid (Daphnia magna)

0.9592

959.2
(Hg/L)

0.014

8,694.5

0.9

(0.5-1.5)

Goldfish (Carassius auratus)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(Ug/L)

<0.0001

13,400

62.2

(31.6-122.5)

Green floater (Lasmigona
subviridis)

Daphnid (Daphnia magna)

0.9592

959.2
(Hg/L)

0.014

166.8

0.9

(0.2-3.5)

Green sunfish (Lepomis
cyanellus)

Rainbow trout
(Oncorhynchus mykiss)

32**

32
(mg/L)

<0.0001

19.93

90.0

(54.2-149.3)

Greenthroat darter
(Etheostoma lepidum)

Rainbow trout
(Oncorhynchus mykiss)

32**

32
(mg/L)

0.004

1.625

34.9

(11-110.9)

Guppy (Poecilia reticulata)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(Ug/L)

0.0006

198.444

28.3

(7.6-105.3)

Isopod (Asellus aquaticus)

Daphnid (Daphnia magna)

0.9592

959.2
(Hg/L)

0.0003

166.8

27.3

(9.3-80.1)

Isopod (Caecidotea
brevicauda)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(Ug/L)

0.0006

82

8.2

(1.9-35.7)

Lake trout (Salvelinus
namaycush)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(Hg/L)

0.0002

198.4

18.0

(11.4-28.4)

Largemouth bass
(Micropterus salmoides)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(Ug/L)

<0.0001

95.86

19.5

(9.5-40.3)

Leech (Nephelopsis obscura)

Daphnid (Daphnia magna)

0.9592

959.2
(Hg/L)

0.0008

4.369

131.2

(18.7-920.2)

Mayfly (Drunella grandis)

Rainbow trout
(Oncorhynchus mykiss)

32A

32
(mg/L)

0.0006

95.86

143.5

(43.1-477.3)

Medaka (Oryzias latipes)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(Hg/L)

0.007

54.400

52.1

(10.9-248.7)

F-72


-------
Predicted Species

Surrogate

ICE Model Value Range
(mg/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Entered
ECso
(units)

Minimum

Maximum

Midge (Chironomus tentans)

Daphnid (Daphnia magna)

0.9592

959.2
(Hg/L)

0.0003

472

4.6

(0.7-32.5)

Midge (Paratanytarsus
dissimilis)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(ug/L)

0.0006

1,330

105.8

(35.6-314.4)

Midge (Paratanytarsus
parthenogeneticus)

Daphnid (Daphnia magna)

0.9592

959.2
(Hg/L)

0.37

14,500

2.9

(0.9-9.1)

Mississippi grass shrimp
(Palaemonetes kadiakensis)

Daphnid (Daphnia magna)

0.9592

959.2
(ug/L)

0.0003

58

0.5

(0.2-1.9)

Mrigal carp (Cirrhinus
mrigala)

Rainbow trout
(Oncorhynchus mykiss)

32**

32
(mg/L)

0.0006

1.625

147.7

(24.4-896.1)

Neosho mucket (Lampsilis
rafinesqueana)

Daphnid (Daphnia magna)

0.9592

959.2
(ug/L)

0.042

166.8

0.8

(0.4-1.6)

Northern leopard frog
(Lithobates pipiens)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(n.g/L)

0.002

198.4

64.7

(33.7-124.2)

Northern pike (Esox lucius)

Rainbow trout
(Oncorhynchus mykiss)

32**

32
(mg/L)

0.005

1.822

18.4

(6.5-52)

Oligochaete (Limnodrilus
hoffmeisteri)

Daphnid (Daphnia magna)

0.9592

959.2
(n.g/L)

0.003

281.6

7.6

(3.1-18.5)

Oligochaete (Tubifex tubifex)

Daphnid (Daphnia magna)

0.9592

959.2
(ug/L)

0.0001

4,894.7

10.8

(3-38.4)

Oriental river shrimp
(Macrobrachium nipponense)

Daphnid (Daphnia magna)

0.9592

959.2
(n.g/L)

0.011

281.6

0.4

(0.2-0.9)

Paper pondshell (Utterbackia
imbecillis)

Daphnid (Daphnia magna)

0.9592

959.2
(ug/L)

0.014

8,694.5

0.8

(0.5-1.3)

Pheasantshell (Ortmanniana
pectorosa)

Daphnid (Daphnia magna)

0.9592

959.2
(n.g/L)

0.042

545.9

0.7

(0.2-2.4)

Rainbow trout
(Oncorhynchus mykiss)*

Daphnid (Daphnia magna)

0.9592

959.2
(ug/L)

0.0001

14,500

1.4
(1.2-1.8)

Razorback sucker
(Xyrauchen texanus)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(n.g/L)

0.003

43.07

26.6

(5.3-134)

F-73


-------
Predicted Species

Surrogate

ICE Model Value Range
(mg/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Entered
ECso
(units)

Minimum

Maximum

Riceland prawn
(Macrobrachium lanchesteri)

Rainbow trout
(Oncorhynchus mykiss)

32**

32
(mg/L)

0.003

2.04

177.6

(34.2-922.7)

Rohu (Labeo rohita)

Rainbow trout
(Oncorhynchus mykiss)

32**

32
(mg/L)

0.0006

1.625

302.0

(61.5-1,484.6)

Shortnose sturgeon
(Acipenser brevirostrum)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(Hg/L)

0.028

95.86

45.7

(11.2-185.7)

Silver perch (Bidyanus
bidyanus)

Rainbow trout
(Oncorhynchus mykiss)

32**

32
(mg/L)

0.001

7.075

40.3

(25.8-62.8)

Snipefly (Atherix variegata)

Rainbow trout
(Oncorhynchus mykiss)

32**

32
(mg/L)

0.0006

0.068

194.9

(89.6-424)

Sockeye salmon
(Oncorhynchus nerka)*

Rainbow trout
(Oncorhynchus mykiss)

32**

32
(mg/L)

0.028

7.5

74.6
(17.1-325.1)

Southern leopard frog
(Lithobates sphenocephalus)

Rainbow trout
(Oncorhynchus mykiss)

32**

32
(mg/L)

0.028

9.7

46.3

(19.9-107.9)

Spotfin chub (Erimonax
monachus)

Rainbow trout
(Oncorhynchus mykiss)

32**

32
(mg/L)

0.004

1.625

24.5

(10.4-57.6)

Swamp lymnaea (Lymnaea
stagnalis)

Daphnid (Daphnia magna)

0.9592

959.2
(Hg/L)

0.014

8,694.5

1.0

(0.4-2.9)

Tadpole physa (Physella
gyrina)

Daphnid (Daphnia magna)

0.9592

959.2
(Ug/L)

0.014

8,694.5

1.0

(0.4-2.3)

Threeridge (Amblema
plicata)

Daphnid (Daphnia magna)

0.9592

0.9592
(mg/L)

0.014

4,894.7

0.3

(0.1-0.7)

Vernal pool fairy shrimp
(Branchinecta lynchi)

Daphnid (Daphnia magna)

0.9592

959.2
(Ug/L)

0.014

8,694.5

0.9

(0.4-1.6)

Walleye (Sander vitreus)

Rainbow trout
(Oncorhynchus mykiss)

32**

32
(mg/L)

0.005

16.24

41.8

(12.1-144.6)

Washboard (Megalonaias
nervosa)

Daphnid (Daphnia magna)

0.9592

959.2
(Ug/L)

0.014

8,694.5

0.4

(0.2-0.9)

Water flea (Chydorus
sphaericus)

Daphnid (Daphnia magna)

0.9592

959.2
(Hg/L)

0.009

977.6

0.4

(0.3-0.7)

F-74


-------
Predicted Species

Surrogate

ICE Model Value Range
(mg/L)

Predicted ECso
(Confidence Limits)
(mg/L)

Species

Measured
ECso
(mg/L)

Entered
ECso
(units)

Minimum

Maximum

Water flea (Moina
macrocopa)

Daphnid (Daphnia magna)

0.9592

959.2
(Hg/L)

0.009

281.6

1.8

(0.6-5.2)

Western pearlshell
(Margaritifera falcata)

Daphnid (Daphnia magna)

0.9592

959.2
(Ug/L)

0.014

8,694.5

0.8

(0.4-1.7)

Western toad (Anaxyrus
boreas)

Rainbow trout
(Oncorhynchus mykiss)

32**

32
(mg/L)

0.028

1.625

31.3

(4.6-211.6)

Yellow perch (Perca
flavescens)

Rainbow trout
(Oncorhynchus mykiss)

32**

32
(mg/L)

<0.0001

16.24

44.5

(21-94.5)

Zebrafish (Danio rerio)

Daphnid (Daphnia magna)

0.9592

959.2
(Hg/L)

0.0001

50.000

3.3b
(1.1-10)

Zebrafish (Danio rerio)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(Ug/L)

0.0002

229.1

17.6

(6.4-48.5)

Zebrafish-embryo (Danio
rerio-embryo)

Daphnid (Daphnia magna)

0.9592

959.2
(Hg/L)

0.0001

46,500

4.8b
(2.5-9.2)

Zebrafish-embryo (Danio
rerio-embryo)

Rainbow trout
(Oncorhynchus mykiss)

32

32,000
(Ug/L)

0.0002

8,341.5

34.4

(16.8-70.4)

*Acceptable models that were not used because genus level empirical data were available.

**Measured ECso falls outside range of ICE model and a "scaled" ECso was therefore entered in ICE model.

A |ig/L CI greater than 50-fold. Used mg/L with acceptable CI.
a Saltwater surrogate species were not used.

b Not used because other more closely taxonomically-related models were available.

0 Not used because other more closely taxonomically-related models where the measured surrogate ECso values fell within the ICE model range were available.

F-75


-------
Table F-23. Ranked 7:3 FTCA Genus Mean Acute Values.

Values in bold and highlighted are derived from empirical 7:3 FTCA toxicity tests with the species.

Rank

MDR

Group1

Common Name

Species

SMAV
(mg/L)

GMAV

Percentile
Rank

1

G

Threeridge

Amblema plicata

0.2500

0.2500

0.02

2

D

Daphnid

Chydorus sphaericus

0.4300

0.4300

0.03

3

G

Washboard

Megalonaias nen'osa

0.4400

0.4400

0.05

4

E

Mississippi grass shrimp

Palaemonetes kadiakensis

0.5300

0.5300

0.07

5

D

Daphnid

Ceriodaphnia dubia

0.6000

0.6000

0.08

6

D

Daphnid

Simocephalus serrulatus

0.6000

0.6527

0.10

D

Daphnid

Simocephalus vetulus

0.7100

7

G

Pheasantshell

Ortmanniana pectorosa

0.7400

0.7400

0.12

8

G

Western pearlshell

Margaritifera falcata

0.7900

0.7900

0.13

9

G

Neosho mucket

Lampsilis rafinesqueana

0.7900

0.8243

0.15

G

Fatmucket

Lampsilis siliquoidea

0.8600

10

G

Paper pondshell

Utterbackia imbecillis

0.8400

0.8400

0.17

11

E

Amphipod

Gammarus fasciatus

0.7600

0.8497

0.18

E

Amphipod

Gammarus pseudolimnaeus

0.9500

12

G

Green floater

Lasmigona subviridis

0.8500

0.8500

0.20

13

E

Vernal pool fairy shrimp

Brcmchinecta Ivnchi

0.8500

0.8500

0.22

14

E

Beaver-tail fairy shrimp

Thamnocephalus platvurus

0.8800

0.8800

0.23

15

G

Tadpole physa

Physella gvrina

0.9500

0.9500

0.25

16

D

Daphnid

Daphnia magna

0.9592

0.9592

0.27

17

G

Swamp lymnaea

Lymnaea stagnalis

1.030

1.030

0.28

18

D

Water flea

Moina macrocopa

1.760

1.760

0.30

19

F

Midge

Paratanytarsus
parthenogeneticus

2.880

2.880

0.32

20

F

Midge

Chironomus teutons

4.630

4.630

0.33

21

H

Oligochaete

Limnodrilus hoffmeisteri

7.620

7.620

0.35

22

E

Isopod

Caecidotea brevicauda

8.170

8.170

0.37

23

E

Riceland prawn

Macrobrachium
lanchesteri

177.6**

8.214

0.38

E

Oriental river shrimp

Macrobrachium
nipponense

0.3800

24

E

Amphipod

Crangonvx pseudogracilis

9.510

9.510

0.40

25

H

Oligochaete

Tubifex tubifex

10.75

10.75

0.42

26

B

Northern pike

Esox lucius

18.36**

18.36

0.43

27

B

Largemouth bass

Micropterus salmoides

19.52

19.52

0.45

28

A

Brook trout

Salvelinus fontinalis

31.44

23.80

0.47

A

Lake trout

Salvelinus namavcush

18.01

29

B

Spotfin chub

Erimonax monachus

24.47**

24.47

0.48

30

B

Zebrafish

Danio rerio

24.57

24.57

0.50

31

B

Razorback sucker

Xvrauchen texanus

26.57

26.57

0.52

32

E

Isopod

Asellus aquaticus

27.30

27.30

0.53

33

B

Guppy

Poecilia reticulata

28.31

28.31

0.55

34

C

Western toad

Anaxvrus boreas

31.29**

31.29

0.57

35

B

Channel catfish

Ictalurus punctatus

31.55

31.55

0.58

36

A

Rainbow trout

Oncorhynchus mykiss

32.00

32.00

0.60

F-76


-------
Rank

MDR

Group1

Common Name

Species

SMAV
(mg/L)

GMAV

Percentile
Rank

37

A

Atlantic salmon

Salmo salar

30.96

32.02

0.62

A

Brown trout

Salmo trutta

33.12

38

B

Cape Fear shiner

Notropis mekistocholas

34.42**

34.42

0.63

39

B

Greenthroat darter

Etheostoma lepidum

34.87**

34.87

0.65

40

B

Common carp

Cyprinus carpio

35.58

35.58

0.67

41

B

Silver perch

Bidvamis bidvanus

40.28**

40.28

0.68

42

B

Walleye

Sander vitreus

41.80**

41.80

0.70

43

B

Yellow perch

Percaflavescens

44.48**

44.48

0.72

44

B

Shortnose sturgeon

Acipenser brevirostrum

45.67

45.67

0.73

45

B

Fathead minnow

Pimephales promelas

46.72

46.72

0.75

46

C

Cuvier's foam froglet

Phvsalaemus cuvieri

51.77

51.77

0.77

47

B

Medaka

Oryzias latipes

52.12

52.12

0.78

48

B

Green sunfish

Lepomis cyanellus

89.98**

54.56

0.80

B

Bluegill

Lepomis macrochirus

33.08

49

C

Bullfrog

Lithobates catesbeianus

58.75

56.04

0.82

C

Northern leopard frog

Lithobates pipiens

64.72

C

Southern leopard frog

Lithobates sphenocephaly

46.28**

50

B

Goldfish

Carassius auratus

62.18

62.18

0.83

51

B

Bonytail

Gila elegans

72.68

72.68

0.85

52

F

Midge

Paratanvtarsus dissimilis

105.8

105.8

0.87

53

H

Leech

Nephelopsis obscura

131.2

131.2

0.88

54

F

Mayfly

Drunella grandis

143.5

143.5

0.90

55

B

Mrigal carp

Cirrhinus mrigala

147 7**

147.7

0.92

56

F

Snipefly

Atherix variegata

194 9**

194.9

0.93

57

B

Colorado squawfish

Ptvchocheilus lucius

195.1

195.1

0.95

58

B

Black bullhead

Ameiurus melas

207.8**

207.8

0.97

59

B

Rohu

Labeo rohita

302.0**

302.0

0.98

**Measured EC50 falls outside range of ICE model and a "scaled" EC50 was therefore entered in ICE model.

1: Freshwater MDR Groups

A)	The family Salmonidae in the class Osteichthyes

B)	A second family in the class Osteichthyes, preferably a commercially or recreationally important

warmwater species (e.g., bluegill, channel catfish, etc.)

C)	A third family in the phylum Chordata (may be in the class Osteichthyes or may be an amphibian, etc.)

D)	A planktonic crustacean (e.g., cladoceran, copepod, etc.)

E)	Abenthic crustacean (e.g., ostracod, isopod, amphipod, crayfish, etc.)

F)	An insect (e.g., mayfly, dragonfly, damselfly, stonefly, caddisfly, mosquito, midge, etc.)

G)	A family in a phylum other than Arthropoda or Chordata (e.g., Rotifera, Annelida, Mollusca, etc.)

H)	A family in any order of insect or any phylum not already represented.

F-77


-------
Table F-24. 7:3 FTCA Final Acute Value and Protective Aquatic Acute Benchmark.

Calculated Freshwater FAY based on 4 lowest values; N=59 GMAVs total





Benchmark calculated by dividing the FAY by 2 and by the carboxylic acid application factor (15.8)





GMAV









Rank

Genus

(mg/L)

ln(GMAV)

ln(GMAV)2

P=R/(N+1)

sqrt(P)

1

Amblema

0.25

-1.39

1.92

0.017

0.129

2

Chydorus

0.43

-0.84

0.71

0.033

0.183

3

Megalonaias

0.44

-0.82

0.67

0.050

0.224

4

Palaemonetes

0.53

-0.63

0.40

0.067

0.258



£ (Sum):

-3.69

3.71

0.17

0.79







P = cumulative probability











R = rank













N = number of GMAVs





S2 =

33.93



S = slope







L =

-2.077



L = X-axis intercept





A =

-0.775



A = InFAV







FAV =

0.4609











FAV/2 =

0.2305 mg/L (Preliminary Value)







Adjustment =

0.2305 / 15.8 = 0.01459 (Preliminary Value / Carboxylic Acid Application Factor)

Benchmark =

0.015 mg/L 7:3 FTCA (rounded to two significant figures)





F-78


-------
0>
>

-2
"3

s

3

-J

J*

s

CB
<&

41

1.00 -r

0.90	V

0.80	-j
0.70

0.60	-¦

o.5o

0.40

g 0.30 4

41

04

0.20
0.10
0.00

Amphibian (WeblCE)
Fish (Empirical)

Fish (WeblCE)

Insect (WeblCE)
Invertebrate (Empirical)
Invertebrate (WeblCE)
Mollusk (WeblCE)
Preliminary Value
¦Acute Benchmark

$

O Labeo
Amehmis
Ptychpcheilus
Athens

O Cirrhinus
+ Drunella
A Nephelopsis
+ Par at anytarsus
O Gila.

J Carassius
Lithobates
Lepomis
Oryzias
Physalaemus
Pimephales
Acipenser
Perca
Sander
_ Bidyanus
1 Cvpnnus
Etheo stoma
Notropis
Safino

Oncorh^Tichjs
Ictalurus
_ Anax\Tus
Poecilia
Asellus
Xyrauchen
Dahio
Erimonax
w Salvelinus
Q Micropterus
O Esox
A Tubifex

A Cianaom'x

i \h -

\lacrobracfium
Caecidotea

		 Limnodrilus

+ Chironomus
+ Paratan\rtarsus
A Moina
O Lymnaea

^ Daphma

.	^ ChySorus

t & |-Am)ble|na| ", , , |

Phy sella
Thamno c ephalus
A Branchinecta
y Lasmigona
A Gammams
O Uttetbackia
O Lampsitis
O ^-arsaritifera

O Ortmahmana
A Simocephalus
A Ceriodaphnia
A Palaemonetes
Aleealonaias

0.01	0.1	1	10	100	1000

Genus Mean Acute Value (mg/L 2H,2H,3H,3H-Pefluorodecanoic acid (7:3 FTC'A))

Figure F-8. Ranked Acute 7:3 FTCA GMAVs Used for the Aquatic Life Acute Benchmark
Calculation.

F.9 Summary of Benchmarks for Evaluated PFAS Substances

Acute freshwater Aquatic Life Ambient Water Quality Benchmarks for the eight selected

PFAS are summarized in Table F-25. These concentrations are expected to be protective of 95%
of freshwater genera exposed to the listed PFAS under short term conditions of one-hour of
duration, if the one-hour average magnitude is not exceeded more than once in three years.
Quantitatively-acceptable empirical toxicity data were coupled with ICE predicted values to
fulfill the eight MDRs for deriving acute freshwater criteria per the Aquatic Life Criteria
Guidelines (U.S. EPA 1985). Using this approach, the eight MDRs were fulfilled for seven of the
evaluated compounds (PFBA, PFHxA, PFNA, PFDA, PFBS, 8:2 FTUCA, and 7:3 FTCA).

F-79


-------
Seven of the eight MDRs were fulfilled for PFHxS and there is considered to be greater
uncertainty associated with this benchmark value compared to benchmarks developed with all
MDRs met. The resulting acute water column-based benchmark magnitudes range from 0.015
mg/L for 7:3 FTC A to 11 mg/L for PFBA.

Table F-25. Acute Freshwater Benchmarks for Eight PFAS.















8:2

7:3

Chemical

PFBA

PFHxA

PFNA

PFDA

PFBS

PFHxS

FTUCA

FTC A

Magnitude1

11

6.0

0.73

0.65

7.6

0.21

0.041

0.015

Duration

One hour average

Frequency

Not to be exceeded more than once in three years on average

1 Values expressed as mg/L, or ppm.

F-80


-------
F.l References

Dowdy, S., S. Wearden and D. Chilko. 2011. Statistics for research. John Wiley & Sons.

Raimondo, S., C. Lilavois and S.A. Nelson. 2024. Uncertainty analysis and updated user
guidance for interspecies correlation estimation (ICE) models and low toxicity compounds.
Integr. Environ. Assess. Manag. Accepted Author Manuscript, https://doi.org/10.1002/ieam.4884

U.S. EPA. (United States Environmental Protection Agency). 1985. Guidelines for deriving
numerical national water quality criteria for the protection of aquatic organisms and their uses.
EPA Report 822/R-85-100 (NTIS Report PB85-227049).

F-81


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Appendix G PFAS Acute Benchmark Calculations using a Data Binning
Approach

The EPA investigated the utility of applying an approach described in Giddings et al.
(2019) for synthetic pyrethroid insecticides as another alternative for deriving benchmark values
for data-limited PFAS. For this approach, Giddings et al. (2019) combined acute toxicity data
available for multiple data-limited synthetic pyrethroid insecticide active ingredients (e.g.,
bifenthrin, cypermethrin, permethrin, etc.) to create a more robust combined species sensitivity
distribution (SSD) with the grouped pyrethroids. Before combining, the available toxicity data
were normalized across the individual pyrethroid active ingredients. This was done by
normalizing the data across the individual pyrethroid active ingredients based on the relative
sensitivity of a single selected "key" species, that has data for each constituent active ingredient.
Giddings et al. (2019) then combined the normalized species data (i.e., key species equivalents)
for the individual pyrethroids and calculated an HCs (hazard concentration at the 5% level) based
on the combined pyrethroid SSD composed of normalized acute toxicity values. Estimated HCss
were then back-calculated for individual constituent pyrethroids based on the relative sensitivity
of the species used for the normalization.

The EPA followed the same methodology to calculate predicted HCs values for selected
PFAS combined into two groups: carboxylic acids and sulfonic acids. Constituent PFAS for
these groups are as follows:

G-l


-------
•	Carboxylic acid PFAS substances for which acceptable acute toxicity data were

available:

o PFOA (perfluorooctanoic acid or perfluorooctanoate)
o PFBA (perfluorobutanoic acid or perfluorobutanoate)
o PFHxA (perfluorohexanoic acid or perfluorohexanoate)
o PFNA (perfluorononanoic acid or perfluorononanoate)
o PFDA (perfluorodecanoic acid or perfluorodecanoate)

•	Sulfonic acid PFAS substances for which acceptable acute toxicity data were available:

o PFOS (perfluorooctane sulfonate or perfluorooctane sulfonate acid)
o PFBS (perfluorobutane sulfonic acid or perfluorobutane sulfonate)
o PFHxS (perfluorohexane sulfonic acid or perfluorohexane sulfonate)

The EPA conducted the following steps to complete this evaluation:

1.	High quality acute toxicity data for each PFAS were compiled. These toxicity data
consist of the same quantitatively acceptable empirical data used for derivation of the
ICE-based values (see Appendix A) for PFBA, PFBS, PFHxA, PFHxS, PFNA,
PFDA, and the same quantitatively acceptable data used for derivation of the PFOA
and PFOS criteria (U.S. EPA 2024a,b). All data were converted into mg/L.

2.	Species mean acute values (SMAVs) were identified. If multiple acute toxicity values
were available for a single species and PFAS substance, then the geometric mean of
those values was calculated.

3.	"Key" species to be used for normalizing the data across the constituent substances
within each PFAS grouping (sulfonic acid PFAS and carboxylic acid PFAS grouping)
were identified and species equivalent values were calculated. Zebrafish (Danio
rerio) was identified as the key species for sulfonic acid PFAS, and the planktonic
crustacean (Daphnia magna) was identified as the key species for carboxylic acid

G-2


-------
PFAS. The SMAV for each constituent substance within each PFAS grouping was
then divided by the corresponding normalization value for the key species to calculate
species equivalent values. The following examples illustrate the procedure for
calculating key species equivalents for daphnids for PFOA within the carboxylic acid
PFAS group:

•	Equivalent for Daphnia magna:

o D. magna PFOA SMAV of 213.9 mg/L I). magna PFOA SMAV of
213.9 mg/L PFOA = 1.00 D. magna equivalent

•	Equivalent for Daphnia pulicaria:

o D. pnlicaria PFOA SMAV of 203.7 mg/L PFOA I). magna PFOA

SMAV of 213.9 mg/L PFOA = 0.9523 D. magna equivalent
The species equivalent values were then ranked. The key species equivalent acute
toxicity values and their rankings are shown in Table G-l for sulfonic acids and
Table G-2 for carboxylic acids.

4. Species equivalent values for constituent substances were pooled for each PFAS
grouping to derive SSDs for sulfonic acid PFAS and for carboxylic acid PFAS. If a
key species equivalent value was available for a species having two or more
constituent PFAS within a group, then the geometric mean of those values was
calculated as the SMAV equivalent for that species. These SMAVs representing key
species equivalent values were then ranked to create SSDs for their respective PFAS
group (sulfonic acid and carboxylic acid PFAS). SSDs were derived for sulfonic acid
PFAS and carboxylic acid PFAS using both the EPA SSD Generator1, as was done by
Giddings et al. (2019), and procedures described in the Aquatic Life Criteria

1 Available online at: Download Software I US EPA. Use of this tool results in the estimation of an HC\.

G-3


-------
Guidelines (U.S. EPA 1985), as is typically done for the derivation of aquatic life
criteria. The SSDs generated by the EPA's SSD Generator are shown in Figure G-la
for sulfonic acid PFAS and Figure G-2a for carboxylic acid PFAS. The SSDs
generated using procedures described in the Aquatic Life Criteria Guidelines are
shown in Figure G-lb for sulfonic acid PFAS and Figure G-2b for carboxylic acid
PFAS.

5. HCs values were calculated for sulfonic acid PFAS and carboxylic acid PFAS from
the SSDs using both the EPA SSD Generator2 and procedures described in the
Aquatic Life Criteria Guidelines (termed Final Acute Value [FAV] for the Aquatic
Life Criteria Guidelines calculations [U.S. EPA 1985]). For any given species within
the PFAS group-specific SSD, a key species equivalent greater than 1.0 indicated it
was less sensitive than the key species, and a value less than 1.0 indicated it was more
sensitive than the key species. Once the key species equivalent HC5 (or FAV) was
calculated, the HC5 (or FAV) species equivalent value calculated from the SSD was
multiplied by the key species toxicity value (i.e., SMAV) for the PFAS of interest
within that group (sulfonic or carboxylic PFAS) to generate a substance-specific HC5
(or FAV). For example:

•	FAV for PFHxS:

o FAV for Sulfonic Acid PFAS of 0.01004 x D. rerio PFHxS SMAV of
22.5 mg/L = PFHxS FAV of 0.2260 mg/L

•	FAV for PFBA:

o FAV for Carboxylic Acid PFAS of 0.008213 xD. magna PFBA SMAV of
4,741 mg/L = PFBA FAV of 38.94 mg/L
The calculations and values are summarized in Tables G-3 through G-6.

2 Available online at: Download Software I US EPA. Use of this tool results in the estimation of an HCs.

G-4


-------
Table G-l. Acute LCsos, SMAVs, normalized Danio rerio equivalent values and rank of SMAV D. rerio equivalent values for
the sulfonic acid PFAS.

folding indicates key species used for normalizing data across constituent substances.				

PFAS

Species

Species Common Name

SMAV3
(mg/L)

Danio rerio
equivalentb

SMAV
D. rerio
equivalent0

Rank of

SMAV
D. rerio
equivalent1*

PFOS

Amby stoma jeffersonianum

Jefferson salamander

51.71

1.86

1.86

17

PFOS

Ambvstoma texanum

Small-mouthed salamander

30.00

1.08

1.08

14

PFOS

Amby stoma tigrinum

Eastern tiger salamander

68.63

2.46

2.46

23

PFOS

Anaxyrus americanns

American toad

56.49

2.03

2.03

18

PFOS

Brachionus calycifloriis

Rotifer

61.80

2.22

2.22

21

PFBS

Danio rerio

Zebrafish

>3,000

1.00

1.00

13

PFHxS

Danio rerio

Zebrafish

22.50

1.00

PFOS

Danio rerio

Zebrafish

27.86

1.00

PFOS

Daphnia carinata

Daphnid

11.56

0.41

0.41

5

PFBS

Daphnia magna

Daphnid

2,183

0.73

1.16

15

PFOS

Daphnia magna

Daphnid

51.86

1.86

PFOS

Daphnia piilicaria

Daphnid

134.0

4.81

4.81

27

PFOS

Dugesia japonica

Planaria

22.48

0.81

0.81

12

PFOS

Elliptio complanata

Eastern elliptio

64.35

2.31

2.31

22

PFOS

Hvla versicolor

Gray treefrog

19.88

0.71

0.71

11

PFOS

Lampsilis siliquoidea

Fatmucket

16.50

0.59

0.59

9

PFBS

Lepomis macrochirns

Bluegill

6,452

2.15

2.15

20

PFOS

Ligumia recta

Black sandshell

13.50

0.48

0.48

6

PFHxS

Lithobates catesbeianus

Bullfrog

1,105

49.11

15.33

30

PFOS

Lithobates catesbeianus

American bullfrog

133.3

4.78

PFOS

Lithobates clamitans

Green frog

113.0

4.06

4.06

25

PFOS

Lithobates pipiens

Northern leopard frog

72.72

2.61

2.61

24

PFOS

Lithobates sylvatica

Wood frog

130.0

4.67

4.67

26

G-5


-------












Rank of











SMAV

SMAV







SMAV3

Danio rerio

D. rerio

D. rerio

PFAS

Species

Species Common Name

(mg/L)

equivalentb

equivalent0

equivalent1*

PFOS

Moinct macrocopct

Daphnid

17.20

0.62

0.62

10

PFOS

Moinct mi crura

Daphnid

0.5496

0.02

0.02

2

PFOS

Neocaridina denticulata

Japanese swamp shrimp

15.61

0.56

0.56

7

PFOS

Neocleon triangulifer

Mayfly

0.07617

0.003

0.003

1

PFOS

Oncorhvnchiis mykiss

Rainbow trout

7.515

0.27

0.27

3

PFOS

Phvsella acuta

Bladder snail

183.0

6.57

6.57

29

PFOS

Physella heterostropha pomilia

Snail

161.8

5.81

5.81

28

PFBS

Pimephales promelas

Fathead minnow

1,938

0.65

0.40

4

PFOS

Pimephales promelas

Fathead minnow

6.950

0.25

PFOS

Pontastacus leptodactylus

Crayfish

48.81

1.75

1.75

16

PFOS

Procambarus fallax f. virginalis

Crayfish

59.87

2.15

2.15

19

PFOS

Xenopus laevis

African clawed frog

15.99

0.57

0.57

8

a Data used for PFBS and PFHxS are presented in Appendix A; data used for PFOS are the same as the quantitatively acceptable data used for derivation of the
PFOS criteria (US EPA 2024b).

b Danio rerio equivalents determined according to the following equation: Species x SMAV for PFAS D. rerio SMAV for PFAS x = D. rerio equivalent for
species.

0 SMAV Danio rerio equivalents are the geometric means of the D. rerio equivalents for that species.
d Rank order of the D. rerio equivalents.

G-6


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Table G-2. Acute LCsos, SMAVs, normalized Daphnia magna equivalent values and rank of SMAV D. magna equivalent
values for carboxylic acid PFAS.

Bolding indicates key species used for normalizing data across constituent substances			

PFAS

Species

Species Common Name

LCso
(mg/L)

SMAV3
(mg/L)

Daphnia
magna
equivalentb

SMAV D.

magna
equivalent0

Rank of

SMAV
D. magna
equivalent1*

PFOA

Ambystoma jeffersonictnum

Jefferson salamander



1,070

5.00

5.00

27

PFOA

Ambystoma texcmum

Small-mouthed salamander



407.3

1.90

1.90

14

PFOA

Ambystoma tigrinum

Eastern tiger salamander



752.0

3.52

3.52

24

PFOA

Anaxyrus americanus

American toad



793.9

3.71

3.71

25

PFBA

Brachionus calycifloriis

Rotifer

110

110.0

0.02

0.13

3

PFHxA

Brachionus ccdycifloriis

Rotifer

140

140.0

0.13

PFOA

Brachionus calycifloriis

Rotifer



150.0

0.70

PFBA

Chydorus sphaericus

Daphnid

>4,280.8

>4,280.8

0.90

0.46

5

PFDA

Chydorus sphaericus

Daphnid

41.13

41.13

0.34

PFNA

Chydorus sphaericus

Daphnid

27.84

27.84

0.33

PFOA

Chydorus sphaericus

Daphnid



93.17

0.44

PFBA

Danio rerio

Zebrafish

>3,000®

13,779

2.91

2.47

18

PFBA

Danio rerio

Zebrafish

13,779

PFOA

Danio rerio

Zebrafish



450.4

2.11

PFOA

Daphnia carinata

Daphnid



66.80

0.31

0.31

4

PFBA

Daphnia magna

Daphnid

>1,006

4,741

1.00

1.00

10

PFBA

Daphnia magna

Daphnid

>4,280.8

PFBA

Daphnia magna

Daphnid

5,251

PFDA

Daphnia magna

Daphnid

81

119.7

1.00

PFDA

Daphnia magna

Daphnid

129.54

PFDA

Daphnia magna

Daphnid

163.48

PFHxA

Daphnia magna

Daphnid

1048

1,048

1.00

PFNA

Daphnia magna

Daphnid

43.42

84.51

1.00

PFNA

Daphnia magna

Daphnid

91.89

G-7


-------














Rank of











Daphnia

SMAV D.

SMAV

PFAS

Species

Species Common Name

LCso
(mg/L)

SMAVa
(mg/L)

magna
equivalentb

magna
equivalent0

D. magna
equivalent1*

PFNA

Daphnia magna

Daphnid

151.29









PFOA

Daphnia magna

Daphnid



213.9

1.00





PFBA

Daphnia piilicaria

Daphnid

>1,006

>1,006

0.21





PFDA

Daphnia piilicaria

Daphnid

149.59

149.6

1.25

0.63

6

PFOA

Daphnia piilicaria

Daphnid



203.7

0.95





PFOA

Dugesia japonica

Planaria



383.6

1.79

1.79

12

PFOA

Hvla versicolor

Gray treefrog



646.2

3.02

3.02

20

PFOA

Lampsilis siliquoidea

Fatmucket



164.4

0.77

0.77

8

PFOA

Lepomis macrochims

Bluegill



664.0

3.10

3.10

21

PFOA

Ligumia recta

Black sandshell



161.0

0.75

0.75

7

PFHxA

Lithobates catesbeiana

Bullfrog

1,105

1,105

1.05

2.24

17

PFOA

Lithobates catesbeiana

American bullfrog



1,020

4.77

PFHxA

Lithobates clamitans

Green frog

758

758.0

0.72

1.90

13

PFOA

Lithobates clamitans

Green frog



1,070

5.00

PFOA

Lithobates pipiens

Northern leopard frog



751.7

3.51

3.51

23

PFOA

Lithobates sylvatica

Wood frog



999.0

4.67

4.67

26

PFOA

Moina macrocopa

Daphnid



166.3

0.78

0.78

9

PFOA

Moina mi crura

Daphnid



0.4747

0.0022

0.0022

1

PFOA

Neocaridina denticulata

Green neon shrimp



431.5

2.02

2.02

15

PFOA

Neocloeon triangulifer

Mayfly



13.05

0.061

0.061

2

PFDA

Oncorhynchus mykiss

Rainbow trout

32

32.00

0.27

2.24

16

PFOA

Oncorhvnchus mvkiss

Rainbow trout



4,001

18.71

PFOA

Physella acuta

Bladder snail



681.1

3.18

3.18

22

PFOA

Pimephales promelas

Fathead minnow



593.6

2.78

2.78

19

PFDA

Xenopus sp.

Clawed frog

76.5

76.50

0.64

1.65

11

PFNA

Xenopus sp.

Clawed frog

335.8

335.8

3.97

G-8


-------
PFAS

Species

Species Common Name

LCso
(mg/L)

SMAV3
(mg/L)

Daphnia
magna
equivalentb

SMAV D.

magna
equivalent0

Rank of

SMAV
D. magna
equivalent1*

PFOA

Xenopus sp.

Frog



377.0

1.76





a Data used for PFBA, PFNA, PFDA, and PFHxA are presented in Appendix A; data used for PFOA are the same as the quantitatively acceptable data used for

derivation of the PFOA criteria (US EPA 2024a).
b Daphnia magna equivalents determined according to the following equation: Species x SMAV for PFAS D. magna SMAV for PFAS x = I), magna

equivalent for species.

0 SMAV Daphnia magna equivalents are the geometric means of the D. magna equivalents for that species.
d Rank order of the D. magna equivalents.
e Non-definitive value, not used in SMAV calculation.

G-9


-------
Table G-3. Results of lognormal regression analysis of combined PFAS group SSDs based
on key species equivalents and using the EPA's SSD Generator.

HC5 value expressed as mg/L.				

PFAS Group

Na

Intercept

Slope

R2

HCs (95% prediction interval)

Sulfonic acidb

30

4.970

1.241

0.827

0.0500 (0.0120-0.2079)

Carboxylic acid0

27

4.946

1.193

0.732

0.04636 (0.00705 - 0.30478)

a Number of species in SSD.

b Danio rerio equivalents.
0 Daphnia magna equivalents.

Table G-4. Calculated freshwater FAV based on four lowest SMAV key species equivalents
for sulfonic acid PFAS.





SMAV









Rank

Species

(mg/L)

ln(SMAV)

ln(SMAV)2

P=R/(N+1)

sqrt(P)

1

Neocloeon triangulifer

0.003

-5.90

34.83

0.032

0.180

2

Moina mi crura

0.02

-3.93

15.41

0.065

0.254

3

Oncorhvnchus mvkiss

0.27

-1.31

1.72

0.097

0.311

4

Pimephales promelas

0.40

-0.91

0.83

0.129

0.359







E(Sum):

-12.05

52.79

0.32

1.10



N

30











S2 =

919.63



S = slope







L =
A =

-11.382
-4.601



L = X-axis intercept
A = InFAV





FAV =

0.01004



P = cumulative probability



Table G-5. Calculated freshwater FAV based on four lowest SMAV key species equivalents
for carboxylic acid PFAS. 				i	





SMAV









Rank

Species

(mg/L)

ln(SMAV)

ln(SMAV)2

P=R/(N+1)

sqrt(P)

1

Moina mi crura

0.0022

-6.11

37.34

0.036

0.189

2

Neocloeon triangulifer

0.061

-2.80

7.82

0.071

0.267

3

Brachionus calvciflorus

0.13

-2.04

4.18

0.107

0.327

4

Daphnia carinata

0.31

-1.16

1.35

0.143

0.378







E(Sum):

-12.11

50.69

0.36

1.16



N

27











S2 =

705.20



S = slope







L =
A =

-10.740
-4.802



L = X-axis intercept
A = InFAV





FAV =

0.008213



P = cumulative probability



G-10


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Table G-6. PFAS acute benchmarks for individual PFAS within sulfonic and carboxylic
acid groups calculated using approach presented by Giddings et al. (2019): Summary of
values determined from SSDs derived using Aquatic Life Criteria Guidelines-based and
SSD Generator-based values.

PFAS

Substance

Group

Aquatic Life Cri
Based Va

teria Guidelines-
ues, mg/L

SSD Generator-Based Values, mg/L

FAV

Benchmark3

HCS,

Benchmark3

PFBS FAV

30.13

15.07

150.0

74.98

PFHxS FAV

0.2260

0.1130

1.125

0.5623

PFOS FAV

0.2798

0.1399

1.393

0.6963

PFBA FAV

38.94

19.47

219.8

109.9

PFNA FAV

0.6941

0.3471

3.918

1.959

PFDA FAV

0.983

0.4916

5.550

2.775

PFHxA FAV

8.607

4.304

48.59

24.29

PFOA FAV

1.757

0.8784

9.917

4.959

a Benchmarks by dividing the FAV or HCs by two, consistent with approach described in the Aquatic Life Criteria
Guidelines (U.S. EPA 1985) for derivation of acute aquatic life criteria.

G-ll


-------
A 1 -

5 0.5 -

Q.

(A

O

C 0.4 -
o

X
o

o 0.3
£

02 -



0 Lithobates catesbeiana
0 Physelia
° PhyseJltflTeterostropfia'pomilia
Daphfrfapulicaria y*

° Litja^oates sylvatica

	ites clam^tans

;' ° Litigates pipiens/'
jystoma tigriotim
. Iiptio complanaia
rachionus caljkiflorus
y'	/Lepomis macfochirus

Procambarus fallax f. virginalis
Anaxyrus imericanus
~ Arnbystqma jeffersonianum

Pontastacus leptodactylus
Daphnia magna
Ambystonia texanum
° Daniorerki)

Ougesia japonica
Hyla versicolor
Moina macrocopa
Lampsilis siliquoidea
Xenopus laevis
5 Neijcaridina denticulata
0 Ligumia recta
° D.aphma carinata
yPimephales promelas
©ncorhynchus mykiss

' Neocleon triangulifer

Stressor Intensity

1.0

0.9

0.8 -¦

0.7

0.6

0.5

0.4

0.3

0.2

0.1

o

Artliropoda

~

Chordata

A

Mollusca

O

Platylielmiiillies

A

Rotifer a

	

Guidelines FAV
SSD Generator HC5

0.0
0.001

O Moina micrura
O Neocleon triangulifer !

|~] Lithobates catesbeiana
A Physelia acuta
A Physelia heterostropha pomilia
O Daphnia pulicaria
~ Lithobates sylvatica
Q Lithobates claniiians
~ Lithobates pipiens
f~] Anibystonia tigrinum
A Elliptio complanata
A Brachionus calyciflorus
Q Lepomis macrochirus
O Procambarus fallax f. virginalis
[~~1 Anaxyrus americanus
Q Ambystonia jeffersonianum
O Pontastacus leptodactylus
O Daplmia magna
~ Ambystoma texanum
~ Daniorerio
O Dugesia japonica
~ Hyla versicolor
O Moina macrocopa
A Lampsilis siliquoidea
~ Xenopus laevis
O Neocaridina denticulata
A Ligumia recta
O Daphnia carinata
Q Pimephales promelas
f~[ Oncorhynchus mykiss

0.01	0.1	1

Species Mean Value (unitless, Danio rerio equivalent)

Figure G-l. Species sensitivity distributions for sulfonic acid PFAS based on Danio rerio equivalents, using the EPA's SSD
Generator (left panel - A) or the Aquatic Life Criteria Guidelines procedure (right panel - B).

G-12


-------
° Ambystoma jeffersonianum
¦*' ° Lithobatessyttatica
° Anaxyru^sfnericanus , ./•''
° AmbysJ^ma tigrinum
Litholwtes pipiens

ella acuta

lomis macrochfrus
fiyla versicolor^
f Pimephales Pamelas
f Danio rerio f
1 Lithobates catesbeiana
• OncorhyncHus mykiss
° Neocaridina denticulata
° Ambystoma texanum
- Lithobates clamitans
° DugeS|i& japonica
« Xenopus sp.
0 Daphnif'magna
• Moina mjfcrocopa
° Lampsjlis siliquoidea
a Liguijita recta
« Dapbrlia pulicaria
° Chydofus sphaericus
' Daphra^ carinata
• Brachiongs ^alyciflorus
Neocloeon triafigulifer

0 -

0.00	0.01	0.10	1.00	10.00

Stressor Intensity

I 10
0.9 -
0.8
i 0.7
I 0.6

u

I

; 0.4
j 0.3
0.2
0.1 -

0.0 •
0.001

o

Arthropoda

~

Chordata

A

Mollusca

O

Platyhelminthes

~

Rotifera

	

Guidelines FAV
SSD Generator HC5

O Moina micrura

Neocloeon triangulifer Q

Ambystoma jeffersonianum ~

Lithobates sylvatica ~

Anaxyras ainericanns ~

Ambystoma tigrinum Q

~ Lithobates pipiens
A Physella acuta
Q Leponus macrochirus
~ Hyla versicolor
~ Pimephales promelas
Q Danio rerio
O Lithobates catesbeiana
~ Oncorhynchus myfciss
O Neocaridina denticulata

~	Ambystoma texanum

~	Lithobates clamitans
O Dtigesia japonica

~ Xenopus sp.

O Dapluiia magna
O Moina macrocopa
A Lampsilis siliquoidea
A Liaumis recta
O Dapluiia pulicaria
O Cliydorus sphaericus
O Daphnia carinata
Brachionus calyci floras

0.01

0.1

Species Mean Value (uniiless, Dapluiia magna equivalent)

Figure G-2. Species sensitivity distributions for carboxylic acid PFAS based on Daphnia magna equivalents, using the EPA's
SSD Generator (left panel - A) or the Aquatic Life Criteria Guidelines procedure (right panel - B)

G-13


-------
G.l References

Giddings, J.M., J. Wirtz, D. Campana and M. Dobbs. 2019. Derivation of combined species
sensitivity distributions for acute toxicity of pyrethroids to aquatic animals. Ecotoxicol. 28: 242-
250.

U.S. EPA (United States Environmental Protection Agency). 1985. Guidelines for deriving
numerical national water quality criteria for the protection of aquatic organisms and their uses.
EPA Report 822/R-85-100 (NTIS Report PB85-227049).

U.S. EPA (United States Environmental Protection Agency). 2024a. Aquatic life ambient water
quality criteria for perfluorooctanoic acid (PFOA). EPA-842-D-22-001. U.S. EPA, Office of
Water, Washington, D.C.

U.S. EPA (United States Environmental Protection Agency). 2024b. Aquatic life ambient water
quality criteria for perfluorooctane sulfonate (PFOS). EPA-842-D-22-002. U.S. EPA, Office of
Water, Washington, D.C.

G-14


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