United States Office of Water EPA-822-R-07-001

Environmental Protection 4304T February 2007

Agency

AQUATIC LIFE AMBIENT 

FRESHWATER QUALITY 

CRITERIA - COPPER

2007 Revision
AQUATIC LIFE AMBIENT FRESHWATER QUALITY CRITERIA - COPPER

2007 Revision

(CAS Registry Number 7440-50-8)

February 2007

U.S. Environmental Protection Agency

Office of Water

Office of Science and Technology

Washington, DC
ii

NOTICES

This document has been reviewed in accordance with U.S. EPA policy and approved for

publication. Mention of trade names or commercial products does not constitute endorsement or

recommendation for use. 

This document can be downloaded from EPA’s website at:

http://www.epa.gov/waterscience/criteria/aqlife.html 
iii
iv

ACKNOWLEDGMENTS

Document Update: 2007

Luis A. Cruz

(document coordinator and contributor)

U.S. EPA

Health and Ecological Effects Criteria

Division

Washington, DC

Cindy Roberts

(contributor)

U.S. EPA

Office of Research and Development

Washington, DC

Mary Reiley

(contributor)

U.S. EPA

Health and Ecological Effects Criteria

Division

Washington, DC

Robert Santore

(contributor)

HydroQual, Inc.

Syracuse, New York

Paul Paquin

(contributor)

HydroQual, Inc.

Syracuse, New York

Gary Chapman

(contributor)

Great Lakes Environmental Center

Columbus, Ohio

Jennifer Mitchell

(contributor)

U.S. EPA (formerly)

Health and Ecological Effects Criteria

Division

Washington, DC

Charles Delos

(contributor)

U.S. EPA

Health and Ecological Effects Criteria

Division

Washington, DC

Joseph Meyer

(contributor)

University of Wyoming

Laramie, Wyoming

Rooni Mathew

(contributor)

HydroQual, Inc.

Syracuse, New York

Tyler K. Linton

(contributor)

Great Lakes Environmental Center

Columbus, Ohio

Statistical Support and Contributor:

Russell Erickson

Office of Research and Development

Environmental Research Laboratory

Duluth, Minnesota
v

CONTENTS

Notices .................................................................... ii

Foreword .................................................................. iii

Acknowledgments ............................................................ iv

Contents ................................................................... v

Acronyms ................................................................. vii

1.0 INTRODUCTION ....................................................... 1

2.0 APPROACHES FOR EVALUATING COPPER BIOAVAILABILITY ............ 2

2.1 General Aspects of Copper Bioavailability .................................. 2

2.2 Existing Approaches .................................................. 4

2.3 The BLM and Its Application to Criteria Development ........................ 5

2.4 BLM Uncertainties and Performance ...................................... 7

3.0 INCORPORATION OF BLM INTO CRITERIA DEVELOPMENT PROCEDURES 11

3.1 General Final Acute Value (FAV) Procedures .............................. 11

3.2 BLM Input Parameters ............................................... 12

3.3 Data Acceptability and Screening Procedures .............................. 12

3.4 Conversion Factors .................................................. 14

3.5 Final Chronic Value (FCV) Procedures ................................... 14

4.0 DATA SUMMARY AND CRITERIA CALCULATION ........................ 14

4.1 Summary of Acute Toxicity to Freshwater Animals and Criteria Calculation .... 14

4.1.1 Comparison with Earlier Hardness-Adjusted Criteria ..................... 16

4.2 Formulation of the CCC ........................................... 17

4.2.1 Evaluation of Chronic Toxicity Data .................................. 17

4.2.2 Calculation of Freshwater CCC ..................................... 18

5.0 PLANT DATA ......................................................... 20

6.0 OTHER DATA ......................................................... 21

7.0 NATIONAL CRITERIA STATEMENT .................................... 22

8.0 IMPLEMENTATION ................................................... 22

9.0 REFERENCES ........................................................ 43
vi

FIGURES

Figure 1. Conceptual Diagram of Copper Speciation and Copper-Gill Model ............... 5

Figure 2. Effects of Increasing Ion Concentration on Acute Lethality To Fathead Minnows .... 9

Figure 3. Comparison of Predicted and Measured Acute Copper Toxicity to P. promelas ..... 10

Figure 4. Ranked Freshwater Genus Mean Acute Values (GMAVs) ..................... 15

Figure 5. Comparison of Hardness Based and BLM Based WQC (Alkalinity and pH Covary

 with Hardness) .................................................... 16

Figure 6. Relationship Between Freshwater Acute Copper Sensitivity (LC50 or EC50) 

 and Acute-Chronic Ratios ............................................. 19

TABLES

Table 1. Acute Toxicity of Copper to Freshwater Animals ........................... 24

Table 2a. Chronic Toxicity of Copper to Freshwater Animals ......................... 34

Table 2b. Chronic Toxicity of Copper to Saltwater Animals ........................... 36

Table 2c. Acute-Chronic Ratios ................................................ 37

Table 3a. Ranked Freshwater Genus Mean Acute Values 

with Species Mean Acute-Chronic Ratios ................................. 38

Table 3b. Freshwater Final Acute Value (FAV) and Criteria Calculations ................. 39

Table 4. Toxicity of Copper to Freshwater Plants.................................. 40

APPENDICES

Appendix A. Ranges in Calibration and Application Data Sets ........................ A-1

Appendix B. Other Data on Effects of Copper on Freshwater Organisms ............... B-1

Appendix C. Estimation of Water Chemistry Parameters for Acute Copper Toxicity Tests . . C-1

Appendix D. Saltwater Conversion Factors for Dissolved Values ..................... D-1

Appendix E. BLM Input Data and Notes ....................................... E-1

Appendix F. Regression Plots ................................................ F-1

Appendix G. Example WQC Values Using the BLM and the Hardness Equation ......... G-1

Appendix H. Unused Data ................................................... H-1
vii

ACRONYMS

ACR Acute-Chronic Ratio

BL Biotic Ligand

BLM Biotic Ligand Model

CCC Criterion Continuous Concentration

CF Conversion Factors

CMC Criterion Maximum Concentration

CWA Clean Water Act

DIC Dissolved Inorganic Carbon

DOC Dissolved Organic Carbon

DOM Dissolved Organic Matter

EC Effect Concentration

EPA Environmental Protection Agency

FACR Final Acute-Chronic Ratio

FAV Final Acute Value

FCV Final Chronic Value

FIAM Free Ion Activity Model

GMAV Genus Mean Acute Value 

GSIM Gill Surface Interaction Model

LC50 Lethal Concentration at 50 Percent Effect Level

LOAEC Lowest Observed Adverse Effect Concentration

NASQAN National Stream Quality Accounting Network

NOAEC No Observed Adverse Effect Concentration

pH Negative logarithm of the concentration (mol/L) of the H3O

+

[H+] ion; scale range

from 0 to 14

SMAV Species Mean Acute Values

STORET EPA STOrage and RETrieval Data System

WER Water-Effect Ratio

WET Whole Effluent Toxicity

WQC Water Quality Criteria
1

1.0 INTRODUCTION

Copper is an abundant trace element found in the earth's crust and is a naturally occurring

element that is generally present in surface waters (Nriagu, 1979). Copper is a micronutrient for

both plants and animals at low concentrations and is recognized as essential to virtually all plants

and animals (Kapustka et al., 2004). However, it may become toxic to some forms of aquatic life at

elevated concentrations. Thus, copper concentrations in natural environments, and its biological

availability, are important. Naturally occurring concentrations of copper have been reported from

0.03 to 0.23 :g/L in surface seawaters and from 0.20 to 30 :g/L in freshwater systems (Bowen,

1985). Copper concentrations in locations receiving anthropogenic inputs can vary anywhere from

levels that approach natural background to 100 :g/L or more (e.g., Lopez and Lee, 1977; Nriagu,

1979; Hem, 1989) and have in some cases been reported in the 200,000 :g/L range in mining areas

(Davis and Ashenberg, 1989; Robins et al., 1997). Mining, leather and leather products, fabricated 

metal products, and electric equipment are a few of the industries with copper-bearing discharges

that contribute to anthropogenic inputs of copper to surface waters (Patterson et al., 1998). 

Over the past 20 years, the U.S. Environmental Protection Agency (EPA) has published a

number of guidance documents containing aquatic life criteria recommendations for copper (e.g.,

U.S. EPA 1980, 1985, 1986, 1996). The present document contains EPA's latest criteria

recommendations for protection of aquatic life in ambient freshwater from acute and chronic toxic

effects from copper. These criteria are based on the latest available scientific information,

supplementing EPA's previously published recommendations for copper. This criteria revision

incorporated new data on the toxicity of copper and used the biotic ligand model (BLM), a metal

bioavailability model, to update the freshwater criteria. With these scientific and technical revisions,

the criteria will provide improved guidance on the concentrations of copper that will be protective

of aquatic life. The BLM is not used in the saltwater criteria derivation because further development

is required before it will be suitable for use to evaluate saltwater data.

This document provides updated guidance to states and authorized tribes to establish water

quality standards under the Clean Water Act (CWA) to protect aquatic life from elevated copper

exposure. Under the CWA, states and authorized tribes are to establish water quality criteria to

protect designated uses. Although this document constitutes EPA's scientific recommendations

regarding ambient concentrations of copper, it does not substitute for the CWA or EPA's

regulations, nor is it a regulation itself. Thus, it cannot impose legally binding requirements on EPA,

states, tribes, or the regulated community, and might not apply to a particular situation based on the

circumstances. State and tribal decision makers retain the discretion in adopting approaches, on a

case-by-case basis, that differ from this guidance when appropriate. EPA may change this guidance

in the future.

Although the BLM has been used in place of the formerly applied hardness-based approach,

the updated freshwater criteria derivations in this document are still based on the principles set forth

in the Guidelines for Deriving Numerical Water Quality Criteria for the Protection of Aquatic Life

and Their Uses (Stephan et al. 1985, hereafter referred to as the Guidelines). Section 2 of this

document provides an overview of copper bioavailability and the BLM. Additional information on

the generalized BLM framework, theoretical background, model calibration, and application for the

BLM can be found in the published literature. Section 3 of this document discusses general
2

procedures and requirements for applying the BLM to criteria. Section 4 provides the derivation of

criteria Final Acute Value (FAV) and Final Chronic Value (FCV) for freshwater organisms. 

Section 5 discusses plant data and Section 6 discusses other data not included in the criteria

derivation. Sections 7 and 8 provide the final criteria statements and information on

implementation. Various supplementary information is provided in several appendices.

2.0 APPROACHES FOR EVALUATING COPPER BIOAVAILABILITY

2.1 General Aspects of Copper Bioavailability

The toxicity of a chemical to an aquatic organism requires the transfer of the chemical from

the external environment to biochemical receptors on or in the organism at which the toxic effects

are elicited. Often, this transfer is not simply proportional to the total chemical concentration in the

environment, but varies according to attributes of the organism, chemical, and exposure

environment so that the chemical is more or less "bioavailable". Definitions of bioavailability vary

markedly (e.g., National Research Council, 2003) and are often specific to certain situations, but a

useful generic definition is the relative facility with which a chemical is transferred from the

environment to a specified location in an organism of interest.

Of particular importance to bioavailability is that many chemicals exist in a variety of forms

(chemical species). Such chemical speciation affects bioavailability because relative uptake rates

can differ among chemical species and the relative concentrations of chemical species can differ

among exposure conditions. At equilibrium in oxygenated waters, "free" copper exists as cupric ion

- Cu(II) weakly associated with water molecules (Cu.nH2O

+2), but this species is usually a small

percentage of the total copper. Most dissolved copper is part of stronger complexes with various

ligands (complexing chemicals that interact with metals), including dissolved organic compounds,

hydroxides, carbonates, and other inorganic ligands. Substantial amounts of copper can also be

adsorbed to or incorporated into suspended particles. More information on copper speciation in

freshwater can be found in Kramer et al. (1997), Bryan et al. (2002), and Smith et al. (2002). 

Copper toxicity has been reported to vary markedly due to various physicochemical

characteristics of the exposure water (e.g., either laboratory or field), including temperature,

dissolved organic compounds, suspended particles, pH, and various inorganic cations and anions,

including those composing hardness and alkalinity (see reviews by Sprague, 1968; Hunt, 1987;

Campbell, 1995; Allen and Hansen, 1996; Paquin et al., 2002). Many of these physicochemical

factors affect copper speciation, and their effects on copper toxicity therefore could be due to

effects on copper bioavailability. That bioavailability is an important factor is evident from uptake

of copper by aquatic organisms being reduced by various organic compounds and inorganic ligands

known to complex copper (Muramoto, 1980; Buckley et al., 1984; Playle et al., 1993 a,b; MacRae

et al., 1999).

A "ligand" is a complexing chemical (ion, molecule, or molecular group) that interacts with a

metal like copper to form a larger complex. A “biotic ligand” is a complexing chemical that is a

component of an organism (e.g. chemical site on a fish gill). For certain ligands, some studies have

demonstrated that the concentration of free copper associated with a specified level of accumulation

or toxicity changes little as the ligand concentration is varied, despite major changes in the
3

proportion of copper bound to the ligand (see review by Campbell, 1995). This suggests that, even

at low concentrations, free copper is more important to bioavailability than the ligand-bound

copper. This is expected if accumulation and toxicity are dependent on the binding of copper to a

biochemical receptor "X" on the surface of the organism, forming a chemical species X-Cu

(receptor-bound metal) that is a first limiting step in accumulation and toxicity. By standard

chemical equilibrium expressions, the amount of such species and the consequent biological effects

would be a function of the activity of just free copper (Morel, 1983 a), a relationship commonly

referred to as the free ion activity model (FIAM). Ligand-bound copper (Cu-L) would contribute

to copper bioavailability if (a) a species X-Cu-L is formed that is important to copper

accumulation/toxicity, (b) the microenvironment near the organism surface is such that Cu-L

dissociates and increases the free copper activity interacting with "X", or (c) copper uptake is via

mechanisms that do not entail binding to such a receptor and can accommodate different copper

species. Some studies have indicated dissolved complexes of copper do contribute to bioavailability

(reviews by Sprague, 1968; Hunt, 1987; Campbell, 1995; Allen and Hansen, 1996; Paquin et al.,

2002).

The effects of physicochemical factors on copper toxicity are diverse and the specific

chemistry of the exposure water will determine whether or not there are appreciable effects on

copper speciation and a resulting strong relationship of toxicity to free copper. Usually copper

toxicity is reduced by increased water hardness (reviews by Sprague, 1968; Hunt, 1987; Campbell,

1995; Allen and Hansen, 1996; Paquin et al., 2002), which is composed of cations (primarily

calcium and magnesium) that do not directly interact with copper in solution so as to reduce

bioavailability. In some cases, the apparent effect of hardness on toxicity might be partly due to

complexation of copper by higher concentrations of hydroxide and/or carbonate (increased pH and

alkalinity) commonly associated with higher hardness. However, significant effects on toxicity

often are still present when hardness is increased in association with anions which do not interact

strongly with copper (Inglis and Davis, 1972; Chakoumakos et al., 1979; Miller and Mackay, 1980;

Erickson et al., 1987). Hardness cations could have some limited effect on copper speciation by

competing with copper for the same dissolved ligands, but increased hardness would then increase

free copper and thus increase, not decrease, toxicity. Sodium has also been reported to affect

copper toxicity (Erickson et al., 1996 b) and pH effects can be partly due to effects of hydrogen ion

other than on copper speciation (Peterson et al., 1984).

The effects of hardness cations could be explained by the competing with copper for the

biochemical receptor "X", thus reducing copper uptake (Zitko, 1976; Zitko et al., 1976; Pagenkopf,

1983). Reduced metal bioavailability due to increased hardness cations has been experimentally

demonstrated (Playle et al., 1992; Meyer et al., 1999, 2002), although this does not specifically

establish cation competition as the mechanism. Pagenkopf (1983) provided a mathematical

description of a Gill Surface Interaction Model (GSIM) that addressed the effects on metal toxicity

of both metal speciation and cations via the interactions of gill surface biochemical receptors with

the free toxic metal, other metal species, hardness cations, and hydrogen ion.

The empirical evidence demonstrates that copper toxicity is affected by exposure conditions

and that much of these effects is plausibly attributed to effects of ligands and cations on copper

bioavailability. However, it should not be presumed that all of the observed effects of the

physicochemical factors on copper toxicity reflect effects on bioavailability, or that bioavailability
4

effects are just due to ligand complexation and cation competition. For example, acute copper

toxicity in aquatic organisms has been related to disruption of osmoregulation, specifically

sodium/potassium exchange (Lauren and MacDonald, 1986; Wood, 1992; Wood et al., 1997;

Paquin et al., 2002), which can be affected by calcium other than by competition with copper for

the same biochemical receptor. Similarly, reported effects of sodium and potassium on copper

toxicity (Erickson et al., 1996 b) might simply reflect favorable or unfavorable ion exchange

gradients, rather than any effect on copper bioavailability. Nevertheless, the effects of ligand

complexation and cation competition on copper bioavailability provide a reasonable conceptual

framework for improved descriptions of how copper toxicity differs across exposure conditions.

2.2 Existing Approaches

EPA aquatic life criteria for metals address the reported effects of hardness on metal toxicity

using empirical regressions of toxic concentrations versus hardness for available toxicity data across

a wide range of hardness (Stephan et al., 1985). Such regressions provided the relative amount by

which the criteria change with hardness, but have certain limitations. The regressions were not just

of hardness, but of any other factor that was correlated with hardness in the toxicity data set used

for the regressions, particularly pH and alkalinity. Although these regressions therefore address

more bioavailability issues than hardness alone, they best apply to waters in which the correlations

among hardness, pH, and alkalinity are similar to the data used in the regressions. The separate

effects of these factors are not addressed for exposure conditions in which these correlations are

different. In addition, some physicochemical factors affecting metal toxicity, such as organic

carbon, are not addressed at all. 

Existing EPA metals criteria also address bioavailability by using dissolved metal as a better

approximation for metal bioavailability than total metal (U.S. EPA, 1993). Although this approach

accounts for the low bioavailability of metal on suspended particles, it does not address the major

effects of various dissolved species on bioavailability. This approach could conceivably be further

developed to include just part of the dissolved copper, but this not only requires resolving what

species to include, how to weight them, and how to assess their concentrations, but also would not

address the effects of cations and other factors that affect toxicity in addition to metal speciation. 

Such a "bioavailable fraction" approach is not justified, because no fraction of metals species

provides a constant measure of toxicity.

To address more completely the modifying effects of water quality than the hardness

regressions achieve, EPA issued guidance in the early 1980s on the water-effect ratio (WER)

method (Carlson et al., 1984; U.S. EPA, 1983, 1992, 1994). The WER is "a biological method to

compare bioavailability and toxicity in receiving waters versus laboratory test waters" (U.S. EPA,

1992). A WER is calculated by dividing the acute LC50 of the metal, determined in water collected

from the receiving water of interest, by the LC50 of the metal determined in a standard laboratory

water, after adjusting both test waters to the same hardness. The standard laboratory water LC50 is

used as the denominator to reflect that this LC50 is measured in test water that has water quality

characteristics representative of the test waters used to develop the Water Quality Criteria (WQC)

toxicity database, at least as a good approximation. The national hardness-based acute criterion

concentration is then multiplied by this ratio (i.e., the WER) to establish a site-specific criterion that

reflects the effect of site water characteristics on toxicity. However, a WER accounts only for
5

interactions of water quality parameters and their effects on metal toxicity to the species tested and

in the water sample collected at a specific location and at a specific time. There is also significant

cost to generate a single WER. 

Because of the limitations of these past approaches for addressing bioavailability in metals

criteria, there is a need for an approach that (1) explicitly and quantitatively accounts for the effect

of individual water quality parameters that modify metal toxicity and (2) can be applied more

cost-effectively and easily, and hence more frequently across spatial and temporal scales. An

assessment framework that incorporates the bioavailability mechanisms discussed in Section 2.1 was

therefore used to address more comprehensively the effects of physicochemical exposure conditions

on copper toxicity with lower costs than required by the WER approach.

2.3 The Biotic Ligand Model and Its Application to Criteria Development

The interactions of toxic metal species and other exposure water constituents with biological

surface receptors described by Zitko (1976), Morel (1983), and Pagenkopf (1983) provided the

basic conceptual and mathematical structure for the bioavailability model to be used here (Figure 1). 

Subsequent experimental work has supported various model tenets by demonstrating the effects of

complexing ligands and competing cations on accumulation of toxic metals at fish gills and the

relationship of toxic effects to accumulation, and has also provided estimates of various model

parameters (Playle et al., 1992, 1993a,b; Janes and Playle, 1995; MacRae et al., 1999, Meyer et al.,

1999, 2002; McGeer et al., 2002). Various efforts in metal speciation modeling also have provided

the ability to do better speciation calculations, especially regarding complexation of metals by

organic matter (e.g., Tipping, 1994). This experimental work has supported further metal toxicity

model development (Meyer, 1999; Brown and Markich, 2000; McGeer et al., 2002; Di Toro et al.,

2001; Santore et al., 2001; Paquin et al., 2002). This bioavailability modeling approach is now

commonly termed “Biotic Ligand Models” to broaden the scope beyond gill surfaces and to

acknowledge that the biochemical receptor "X" discussed in Section 2.1 is a metal-binding ligand

that is treated similarly to ligands in the exposure water, except that it is on the organism and is the

keystone for metal accumulation and toxicity. 
6

Equation 1

Equation 2

Equation 3

Equation 4

Briefly, available evidence indicates that both free copper and copper monohydroxide bind to a

biotic ligand "Lb" on the organism's surface (Lb-Cu and Lb-CuOH) and that death occurs when a

certain amount of the total biotic ligand sites are occupied by copper. This ligand must be at the

organism surface because the model describes its interactions with the external exposure water. 

However, this does not mean that this ligand is the site of toxic action; rather it is only necessary to

assume that copper accumulation at the site(s) of toxic action is proportional to binding at the biotic

ligand (i.e., the biotic ligand controls bioavailability). Other cations also will bind to the biotic

ligand, affecting copper bioavailability because higher concentrations of copper are needed for

copper to reach toxic levels. The binding to the biotic ligand is considered to be at equilibrium,

with apparent (activity-corrected) equilibrium constants KLbCu, KLbCuOH, and KLbCj, respectively, for

free copper, copper hydroxide, and the "jth" competing cation. Chemical speciation in the exposure

water is also considered to be at equilibrium, and chemical speciation calculations are conducted to

compute the free copper, copper hydroxide, and competing cation activities to which the biotic

ligand is exposed. Because binding to the actual biotic ligand cannot be measured, it is expected

that accumulation relationships for some measurable variable (e.g., the total metal in gill tissue)

provide a reasonable surrogate for the actual biotic ligand. Because criteria deal with

concentrations eliciting a certain level of effects on groups of organisms (e.g., LC50s), model

calculations are for an organism with characteristics appropriate for such group-wide statistics. 

How the BLM is applied to criteria can be best discussed by starting with the following

general expression for the BLM:

where EC is the total dissolved copper concentration eliciting an effect, EC0

 is a baseline EC in the

absence of any complexing ligands and competing cations, fC should be a factor (<1) for how much

competing cations increase EC, and fL should be a factor (<1) for how much complexing ligands

increase EC. For the BLM used here: 

where fLbT is the fraction of the biotic ligand sites that must be occupied by copper to elicit the

toxicity of interest (e.g., a lethal accumulation divided by the accumulation capacity), m is the
7

Equation 5

number of competing cations included in the model, [Cj] is the concentration of the jth competing

cation, "Cu+2 is the ratio of free copper concentration to total dissolved copper concentration, "CuOH

is the ratio for the copper hydroxide complex, and the ratio KLbCuOH/KLbCu specifies the

bioavailability of CuOH relative to free copper. Thus, in the absence of complexing ligands and

competing cations, the toxic concentration is only a function of the binding strength of free copper

and the copper occupied fraction of biotic ligand sites needed to elicit toxicity. The increase in the

effect concentration due to competing cations is simply a sum of the products of their

concentrations and binding constants. The increase in the effect concentration due to complexing

ligands is the inverse of the sum of the products of the relative bioavailabilities and concentration

fractions of the species that bind to the biotic ligand (free copper and copper hydroxide).

If toxicity to all the biological species in the criteria (at least the most sensitive ones) were

determined based on measured accumulation properties and the relationship of toxicity to

accumulation, the above model equations would be directly applied in criteria calculations. 

However, this is not the case. Although gill accumulation properties and lethal accumulations have

been measured for certain species and conditions, and this has been useful in validating BLM

assumptions and formulations, the data that must be applied to the criteria consists of water effect

concentration (ECs) for biological species for which this accumulation information is generally not

available. The BLM therefore is needed, not to make absolute calculations regarding toxic

concentrations, but to extrapolate toxic concentrations from one exposure condition to another:

where the A and B subscripts refer to different exposure conditions. The general procedure that

was followed for criteria development here was to use the above equation to normalize all available

toxicity data to a reference exposure condition, calculate criteria values at the reference condition,

and again use the above equation to compute criteria at other conditions. 

This means that the BLM assumptions and parameters that just pertain to EC0

 are not

important to its application to criteria, which actually simplifies model validation and

parameterization needs. In particular, there is no need to estimate fLbT, or the lethal accumulations

and accumulation capacities that define this fraction. Furthermore, the absolute values of KLbCu and

KLbCuOH do not need to be known, only their relative value (and if copper binding to the biotic ligand

was dependent only on free copper, the value of KLbCu would not be needed at all). Absolute values

are only needed for the binding constants for the competing cations, as well as the various constants

needed in speciation calculations to estimate "Cu+2 and "CuOH. For BLM application to criteria, the

important concern is whether fC

 and fL

 are suitably formulated and parameterized, and not with

issues that relate to lethal accumulations and accumulation capacities.

2.4 BLM Uncertainties and Performance

The BLM employed here uses equilibrium reactions of copper and other cations with a single,

simple type of surface ligand as the focus for all the effects of physicochemical exposure conditions

on toxicity, and thus is a simple, approximate representation for the complex set of chemical
8

reactions and transfers involved with environmental copper concentrations eliciting toxicity. As

already noted, cation effects might involve mechanisms other than competition for a surface ligand. 

The microenvironment at the gill might change copper speciation. Multiple mechanisms that do not

react the same to external conditions might be involved in copper bioavailability and toxicity. 

Accumulation parameters based on bulk gill measurements will likely not be the same as those for

the biotic ligand. Nonequilibrium processes might be important, especially regarding the

relationship of copper-binding on a surface ligand to toxic action.

However, any model is a simplification of reality and the existence of uncertainties does not

preclude a model from being useful and justified. Despite its simplicity, the BLM used here

provides a reasonable mechanistic framework for the well-established effects of copper speciation,

explicitly addressing the relative bioavailability of different copper species. It also includes a

plausible mechanism that allows the effects of cations to be addressed and uses a comprehensive

model for calculating the required concentrations of various chemical species. Even if the

mechanistic descriptions are incomplete, this model allows the major empirical effects of

complexing ligands and competing cations to be described in a more comprehensive and reasonable

fashion than other approaches. 

Because this model is used in criteria to predict relative effects of physicochemical exposure

factors, its utility for criteria can be judged based on how well it predicts the relative effects of these

factors in copper toxicity studies. Examples of BLM performance for various exposure factors and

studies are provided in the technical support document for this criteria. Figure 2 shows one

example from a study on the effects of various exposure conditions on the acute lethality of copper

to fathead minnows. This set of exposures consisted of synthetic exposure solutions of various

total ion concentrations with fixed ratios of the major cations and anions, at a fixed pH (8.0) and

low dissolved organic matter (< 0.5 mg/L). Observed dissolved LC50s (solid circles with

uncertainty bars) varied by 24-fold for only a 9-fold change in total ions. These large effects reflect

the combined influences of increased alkalinity (copper carbonate complex formation), hardness,

and sodium. Considering the wide range of the observed LC50s and that the model was not fitted

to these data, BLM-predicted LC50s (open symbols) were rather accurate, ranging from 55 to 87%

(average 75%) of the observed value. More importantly for criteria, the predicted relative change

across the range of total ion concentration was 20-fold, very close to that observed.
9

Model performance can also be judged across a variety of factors as in Figure 3, which shows

predicted versus observed LC50s for a large number of exposures in the cited study, which varied

hardness, alkalinity, sodium, and pH together and separately over a wide range. Observed LC50s

varied by about 60-fold, but predicted values deviated from observed values by only 0.12 log units

(a factor of 1.3) on average, and at worst only slightly more than a factor of 2. Again, more

information on model performance is provided in the Technical Support Document and the figures

here just provide some examples demonstrating the utility of this model for use in criteria.
10

The use of the BLM to predict the bioavailability and toxicity of copper to aquatic organisms

under site-specific conditions is a significant change from the previous Criterion Maximum

Concentration (CMC) derivation methodology. Previous aquatic life criteria documents for copper

(e.g., U.S. EPA, 1980, 1985, 1996) expressed the CMC as a function of water hardness. Now, EPA

chooses to utilize the BLM to update its freshwater acute criterion because the BLM accounts for

all important inorganic and organic ligand interactions of copper while also considering competitive

interactions that influence binding of copper at the site of toxicity, or the "biotic ligand." The BLM's

ability to incorporate metal speciation reactions and organism interactions allows prediction of

metal effect levels to a variety of organisms over a wide range of water quality conditions.

Accordingly, the BLM is an attractive tool for deriving water quality criteria. Application of the

BLM has the potential to substantially reduce the need for site-specific modifications, such as Water

Effect Ratio, to account for site-specific chemistry influences on metal toxicity.

The updated BLM-based WQC will in some cases be more stringent and in other cases less

stringent than the hardness based WQC. As there is not a single WQC value to use for comparison

purposes, it will only be possible to provide illustrative examples of each situation. It is the 

judgement of the EPA that the BLM-based WQC for Cu will provide an improved framework for

evaluating a level of protection (LOP) that is consistent with the LOP that was intended by the

1985 Guidelines (i.e., a 1-in-3 year exceedance frequency that will be protective of 95% of the

genera).

While the BLM is currently considered appropriate for use to derive an updated freshwater

CMC for the acute WQC, further development is required before it will be suitable for use to

+2

-2
11

Equation 6

Equation 7

evaluate a saltwater CMC or a Criterion Continuous Concentration (CCC) or chronic value

(freshwater or saltwater WQC).

3.0 INCORPORATION OF THE BLM INTO CRITERIA DERIVATIONS 

PROCEDURES

3.1 General Final Acute Value (FAV) Procedures

Application of the acute copper BLM to the derivation of the copper FAV is analogous to

procedures already described in the Guidelines for metals criteria using empirical hardness

regressions. For these hardness-dependent metals criteria, LC50s at various hardness are

normalized to a reference hardness using the regression slopes. The normalized LC50s for each

biological species are averaged to derive Species Mean Acute Values (SMAVs) at the reference

hardness. The SMAVs within each genus are then averaged to derive Genus Mean Acute Values

(GMAVs) at the reference hardness. The Guidelines’ procedures for estimating the fifth percentile

of the GMAVs are then used to derive the FAV at the reference hardness. FAVs for other hardness

can then be derived using the hardness regression slope, and these FAVs are used to calculate the

Criterion Maximum Concentration (CMC) by dividing the FAV by 2.0 and the Final Chronic Values

(FCV) by dividing the FAV by the Final Acute-Chronic Ratio (FACR). Following the Guidelines,

the Criterion Continuous Concentration (CCC) is set to the FCV unless other data justifies a lower

value.

Extending this procedure to apply the BLM simply involves normalizing the LC50s to a

reference exposure condition that includes all the physicochemical exposure factors important to the

BLM, not just hardness. For this normalization, the BLM provides the factors fC

 and fL

 discussed

in Section 2.3, these factors serving the same purpose as the hardness regression slope described

above. Each LC50 to be used in criteria derivation would be normalized to the reference exposure

conditions by the equation:

where the subscript A refers to the exposure conditions for the observed LC50 and the subscript R

refers to the reference exposure conditions to which the LC50 is being normalized. These

normalized LC50s are then used to derive the SMAVs, GMAVs, and FAV at the reference

exposure condition as described above for the hardness-corrected criteria. The BLM is then used

to derive FAVs at other exposures by the equation:
12

where the subscript B refers to the exposure conditions for which an FAV is desired. These

BLM-derived FAVs are then used to derive CMCs and CCCs following standard Guidelines

procedures.

For the criteria in this document, the reference exposure conditions to which LC50s are

normalized and at which the reference FAV is calculated are as follows (see also footnote f in Table

1). The water chemistry used in the normalization was based on the EPA formulation for

moderately-hard reconstituted water, but any other water chemistry could have been used. In this

formulation the parameters included: temperature = 20oC, pH = 7.5, DOC = 0.5 mg/L, Ca = 14.0

mg/L, Mg = 12.1 mg/L, Na = 26.3 mg/L, K = 2.1 mg/L, SO4 =81.4 mg/L, Cl = 1.90 mg/L, 

Alkalinity = 65.0 mg/L and S = 0.0003 mg/L.

3.2 BLM Input Parameters

For applying an LC50 to criteria derivations and for determining an FAV at exposure

conditions of interest, the necessary water quality input parameters for BLM calculations are

temperature, pH, dissolved organic carbon, major geochemical cations (calcium, magnesium,

sodium, and potassium), dissolved inorganic carbon (DIC, the sum of dissolved carbon dioxide,

carbonic acid, bicarbonate, and carbonate), and other major geochemical anions (chloride, sulfate). 

DIC measurements are typically not made in the environment, and an alternative input parameter is

alkalinity, which can be used with pH and temperature to estimate DIC. There is some evidence

that other metals such as iron and aluminum can have an effect on copper toxicity to aquatic

organisms, which might be due to interactions of these metals with the biotic ligand, effects of these

metals on organic carbon complexation of copper, or adsorption of copper to iron and aluminum

colloids which are present in filtrates used to measure dissolved copper. These metals are not

currently included in routine BLM inputs, but users are encouraged to measure dissolved iron and

aluminum as part of monitoring efforts to support possible future criteria applications.

A number of fixed parameters are also used in the BLM but are not required user inputs in

criteria derivations. These include the variety of equilibrium constants used in copper speciation

calculations, and also the binding constants for copper and various cations to the biotic ligand. The

values for these constants were obtained from work by Playle and coworkers (Playle et al., 1992,

1993a,b) and also by inference from the relationship of toxicity to various water quality

characteristics. More information about these parameters can be obtained from the technical

support document. 

3.3 Data Screening Procedures

To use a toxicity test in the derivation of BLM-based criteria, information must be available 

for the various water quality parameters described in Section 3.2. This is in contrast to past metals

criteria, for which the only necessary water quality parameter was hardness. Many of these

parameters are not routinely measured in toxicity tests and, if measured, are not necessarily

reported in the primary literature for the test, especially for older toxicity tests. However, this

information might be available from supplemental sources or be estimated based on other

information. Therefore, in addition to reviewing the primary sources for relevant information,
13

additional efforts were made to obtain or estimate the necessary water quality parameters for as

many of the available LC50s as possible.

A detailed description of these efforts is provided in Appendix C, Estimation of Water

Chemistry Parameters for Acute Copper Toxicity Tests, and are summarized as follows. Reports of

acute copper toxicity tests identified in literature searches were reviewed to identify LC50s for

possible inclusion in the criteria derivation. In addition to test acceptability standards specified in

the Guidelines, the current effort also required that the LC50s be based on measured copper

concentrations. LC50s based on nominal concentrations have been used in previous criteria, but

there are enough measured LC50s for copper that this was considered to be no longer warranted,

especially considering the more advanced bioavailability assessments represented by the BLM. For

the identified LC50s, the primary reports were reviewed to record all reported information on

dilution and test water chemistry. Any additional references specified by the authors were also

obtained and reviewed. If test waters were synthetically prepared based on specified formulas,

these were used to estimate parameters as appropriate. When critical water chemistry parameters

were not available, authors were contacted regarding unpublished information or to measure

missing water chemistry parameters in dilution source waters. If primary or corresponding authors

could not be contacted, an attempt was made to contact secondary authors or personnel from the

laboratories where the studies were conducted. Where actual water chemistry data were

unavailable, data from other studies with the same water source were used as surrogate values if

appropriate. Absent this, the U.S. Geological Survey's National Stream Quality Accounting

Network (NASQAN) and the EPA STOrage and RETrieval (STORET) were used to obtain data

for ambient surface waters which were the source of water for a test. In some instances other

available sources were contacted to obtained water chemistry data (e.g., city drinking water

treatment personnel). The acquired data were scrutinized for representativeness and usefulness for

estimating surrogate values to complete the water quality information for the dilution and/or test

water that was used in the original studies. When the above sources could not be used,

geochemical ion inputs were based on reported hardness measurements and regressions

relationships constructed for the relationship of various ions to hardness from NASQAN data.

As with any modeling effort, the reliability of model output depends on the reliability of model

inputs. Although the input data have been closely scrutinized, the reliability of the BLM-normalized

LC50s are subject to the uncertainties of the estimation procedures described above. Therefore, a

ranking system was devised to rank the quality of the chemical characterization of the test water. 

Studies with a rank of 1 contain all of the necessary parameters for BLM input based on

measurements from either the test chambers or the water source. In general, studies in which the

BLM input parameters were reported for test chamber samples take precedence over studies in

which the parameters were reported only for the source water. A characterization ranking of 2

denotes those studies where not all parameters were measured, but reliable estimates of the

requisite concentrations could be made. Similarly, a rank of 3 denotes studies in which all

parameters except DOC were measured, but reliable estimates of DOC could be made. For the

majority of the tests, a chemical characterization of 4+ was assigned because hardness, alkalinity,

and pH were measured, and the ionic composition could be reliably estimated or calculated. A 4-

was assigned to those studies conducted using standard reconstituted water in which hardness,

alkalinity, or pH was either measured or referenced, and the recipe for the water is known (ASTM,

2000; U.S. EPA, 1993). The chemical characterization rank of 5 was ascribed to studies in which
14

one of the key parameters (DOC, Ca, pH, alkalinity) was not measured, and when it could not be

reliably estimated. If two or more key parameters (DOC, Ca, pH, alkalinity) were not measured and

could not be reliably estimated, a study was given a chemical characterization rank of 6. Studies

receiving a quality rating of greater than 4+ (i.e., higher than 4) were not used in the criteria

development procedures because the estimates for some of the key input parameters were not

thought to be reliable, all other studies were used.

3.4 Conversion Factors

The LC50s used in deriving previous EPA metals criteria were based on total metal

concentration (measured or nominal) and the criteria were consequently for total metals

concentration. EPA afterwards made the decision that metals criteria should be based on dissolved

metal because it was thought to better represent the bioavailable fraction of the metal (U.S. EPA,

1993). It was thus necessary to convert the criteria to a dissolved concentration basis. However, at

that time, most toxicity tests reported only total concentration, so that a procedure was necessary to

estimate the likely fractions of metals that were dissolved in typical toxicity tests. Studies were

therefore conducted to determine these fractions under a variety of test conditions that mimicked

the conditions in the tests used to derive the metals criteria (University of Wisconsin-Superior,

1995). These tests demonstrated high fractions of dissolved copper and resulted in a conversion

factor (CF) of 0.96 for converting both the CMC and CCC for copper from a total to dissolved

basis (Stephan, 1995). The BLM-derived criteria developed here also uses dissolved copper as the

basis for criteria, assuming a negligible bioavailability for particulate copper. The conversion factor

of 0.96 was also used to convert total to dissolved copper for any toxicity test for which dissolved

copper measurements were not available.

3.5 Final Chronic Value (FCV) Procedures

Because the minimum eight family data requirements for chronic toxicity data were not met in

order to calculate the FCV by the fifth percentile method used for the FAV and because insufficient

information was available to develop a chronic BLM, EPA derived the CCC utilizing the Acute to

Chronic Ratio (ACR) approach from the Guidelines (Stephan et al., 1985). To calculate the FCV at

a specific water chemistry, the FAV at that chemistry is divided by the FACR. This entails the

assumption that the acute BLM reasonably approximates the bioavailability relationships for chronic

toxicity. Limited data available regarding effects of water chemistry on sublethal effects and

chronic lethality do show substantial effects of organic matter, alkalinity, pH, and sodium (Winner,

1985; Erickson et al., 1996 a,b) similar to those in the acute BLM used here. For hardness,

apparent effects are limited and uncertain, but the use of the acute BLM does not introduce major

uncertainties in this regard because the effects of hardness by itself in the acute BLM are also

limited.

4.0 DATA SUMMARY AND CRITERIA CALCULATION

4.1 Summary of Acute Toxicity to Freshwater Animals and Criteria Calculation

The screening procedure outlined in Sec. 3.3 (high quality data = 1, low quality data > 4, e.g.

4+) identified approximately 600 acute freshwater toxicity tests with aquatic organisms and copper
15

potentially acceptable for deriving criteria. Of these tests, approximately 100 were eliminated from

the criteria derivation process because they did not report measured copper concentrations. Nearly

150 additional tests were eliminated from the calculation of the FAV because they received a quality

rating of greater than 4 in the quality rating scheme described in section 3.3 described above. 

Data from approximately 350 tests were used to derive normalized LC50 values, including 15

species of invertebrates, 22 species of fish, and 1 amphibian species (Table 1), representing 27

different genera. Species Mean Acute Values (SMAVs) at the reference chemistry were calculated

from the normalized LC50s and Genus Mean Acute Values (GMAVs) at the normalization

chemistry were calculated from the SMAVs. 

SMAVs ranged from 2.37 μg/L for the most sensitive species, Daphnia pulicaria, to 107,860

μg/L for the least sensitive species, Notemigonus crysoleucas. Cladocerans were among the most

sensitive species, with D. pulicaria, D. magna, Ceriodaphnia dubia, and Scapholeberis sp. being

four out of the six most sensitive species. Invertebrates in general were more sensitive than fish,

representing the 10 lowest SMAVs.

The 27 GMAVs calculated from the above-mentioned SMAVs ranged from 4.05 μg/L for

Daphnia to 107,860 μg/L for Notemigonus (Table 3a). Nine of the 10 most sensitive genera were

invertebrates. The salmonid genus Oncorhynchus was the most sensitive fish genus, with a GMAV

of 31.39 μg/L and an overall GMAV ranking of 10. 

The ranked GMAVs are presented in Figure 4. Pursuant to procedures used to calculate the

FAV, a FAV of 4.67 :g/L was derived from the four GMAVs with cumulative probabilities closest

to the 5th percentile toxicity value for all the tested genera (Table 3b). The presumption is that this 

Figure 4. Ranked Freshwater Genus Mean Acute Values (GMAVs)
16

acute toxicity value represents the LC50 for an organism that is sensitive at the 5th percentile of the

GMAV distribution. The CMC is the FAV divided by two. Therefore, the freshwater dissolved

copper CMC for the reference chemistry presented is 2.337 μg/L.

Site-water chemistry parameters are needed to evaluate a criterion. This is analogous to the

situation that previously existed for the hardness-based WQC, where a hardness concentration was

necessary in order to derive a criterion. Examples of CMC calculations at various water chemistry

conditions are presented in Figure 5 and Appendix G.

4.1.1 Comparison With Earlier Hardness-Adjusted Criteria

EPA’s earlier freshwater copper criteria recommendations were hardness-dependent values.

One would expect a BLM-based criterion calculation procedure to yield the more appropriate

criterion—appropriate in the sense that it accounts for the important water chemistry factors that

affect toxicity, including DOC complexation, where the hardness correction does not. Application

of the BLM in field situations where DOC is expected to be present at higher concentrations than

those observed in laboratory studies would likely improve the performance of the BLM compared

with the hardness adjustment. The reason is that the BLM would reasonably account for the

typically observed increase in effect levels under such conditions, while the hardness-based

approach would not (Figure 5).

As a comparison between the hardness typical of the previous copper criterion and this revised

criterion using the BLM, both procedures were used to calculate criterion values for waters with a

range in hardness as specified by the standard EPA recipes (U.S. EPA, 1993). The EPA

formulations specify the concentration of various salts and reagents to be used in the synthesis of
17

laboratory test waters with specific hardness values (e.g., very soft, soft, moderately hard, hard, or

very hard). As the water hardness increases in these recipes, pH and alkalinity also increase. This

has implications for the BLM because the bioavailability of copper would be expected to decrease

with increasing pH and alkalinity due to the increasing degree of complexation of copper with

hydroxides and carbonates and decreasing proton competition with the metal at both DOM and

biotic ligand binding sites. The BLM criterion for these waters agrees very well with that calculated

by the hardness equation used in previous copper criterion documents (Figure 5). However,

alkalinity and pH change as hardness changes in the EPA recipes. The BLM prediction is taking all

of these changes in water quality into account. 

It is possible to use the BLM to look only at the change in predicted WQC with changes in

hardness (e.g., alkalinity and pH remaining constant). The hardness equation is based on waters

where changes in hardness are accompanied by changes in pH and alkalinity. However, there are

many possible natural waters where changes in hardness are not accompanied by changes in pH and

alkalinity (such as water draining a region rich in gypsum). In these cases, the hardness equation

based criterion will still assume a response that is characteristic of waters where hardness, alkalinity,

and pH co-vary, and will likely be underprotective relative to the level of protection intended by the

Guidelines, in high hardness waters. Conversely, in waters where the covariation between hardness,

pH, and alkalinity is greater than is typical for data in Table 1, the hardness equation based criteria

may be overprotective. Appendix G shows representative water quality criteria values using both

the BLM and the hardness equation approaches for waters with a range in pH, hardness, and DOC

concentrations. The hardness approach does not consider pH and DOC while the BLM approach

takes those water quality parameters into consideration.

4.2 Formulation of the CCC

4.2.1 Evaluation of Chronic Toxicity Data

In aquatic toxicity tests, chronic values are usually defined as the geometric mean of the

highest concentration of a toxic substance at which no adverse effect is observed (highest no

observed adverse effect concentration, or NOAEC) and the lowest concentration of the toxic

substance that causes an adverse effect (lowest observed adverse effect concentration, or LOAEC).

The significance of the observed effects is determined by statistical tests comparing responses of

organisms exposed to low-level and control concentrations of the toxic substance against responses

of organisms exposed to elevated concentrations. Analysis of variance is the most common test

employed for such comparisons. This approach, however, has the disadvantage of resulting in

marked differences between the magnitudes of the effects corresponding to the individual chronic

values, because of variation in the power of the statistical tests used, the concentrations tested, and

the size and variability of the samples used (Stephan and Rogers, 1985). 

An alternative approach to calculating chronic values focuses on the use of point estimates

such as from regression analysis to define the dose-response relationship. With a regression

equation or probit analysis, which defines the level of adverse effects as a function of increasing

concentrations of the toxic substance, it is possible to determine the concentration that causes a

specific small effect, such as a 5 to 30 percent reduction in response. To make chronic values reflect

a uniform level of effect, regression and probit analyses were used, where possible, both to

demonstrate that a significant concentration-effect relationship was present and to estimate chronic
18

values with a consistent level of effect. The most precise estimates of effect concentrations can

generally be made for 50 percent reduction (EC50); however, such a major reduction is not

necessarily consistent with criteria providing adequate protection. In contrast, a concentration that

causes a low level of reduction, such as an EC5 or EC10, might not be statistically significantly

different from the control treatment. As a compromise, the EC20 is used here to represent a low

level of effect that is generally significantly different from the control treatment across the useful

chronic datasets that are available for copper. The EC20 was also viewed as providing a level of

protection similar to the geometric mean of the NOEC and LOEC. Since the EC20 is not directly

dependent on the tested dilution series, similar EC20s should be expected irrespective of the tested

concentrations, provided that the range of tested concentrations is appropriate.

Regression or probit analysis was utilized to evaluate a chronic dataset only in cases where the

necessary data were available and the dataset met the following conditions: (1) it contained a

control treatment (or low exposure data point) to anchor the curve at the low end, (2) it contained

at least three concentrations, and (3) two of the data points had effect variable values below the

control and above zero (i.e., “partial effects”). Control concentrations of copper were estimated in

cases where no measurements were reported. These analyses were performed using the Toxicity

Relationship Analysis Program software (version 1.0; U.S. EPA, Mid-Continental Ecology

Division, Duluth, MN, USA). Additional detail regarding the aforementioned statistical procedures

is available in the cited program. 

When the data from an acceptable chronic test met the conditions for the logistic regression or

probit analysis, the EC20 was the preferred chronic value. When data did not meet the conditions

the chronic value was usually set to the geometric mean of the NOAEC and the LOAEC. However,

when no treatment concentration was an NOAEC, the chronic value is reported as less than the

lowest tested concentration.

For life-cycle, partial life-cycle, and early life stage tests, the toxicological variable used in

chronic value analyses was survival, reproduction, growth, emergence, or intrinsic growth rate. If

copper apparently reduced both survival and growth (weight or length), the product of variables

(biomass) was analyzed, rather than analyzing the variables separately. The most sensitive of the

toxicological variables was generally selected as the chronic value for the particular study.

A species-by-species discussion of each acceptable chronic test on copper evaluated for this

document is presented in Appendix F. Figures that present the data and regression/probability

distribution line for each of the acceptable chronic test which contained sufficient acceptable data

are also provided in Appendix F.

4.2.2 Calculation of Freshwater CCC

Acceptable freshwater chronic toxicity data from early life stage tests, partial life-cycle tests,

and full life-cycle tests were available for 29 tests including data for 6 invertebrate species and 10

fish species (Table 2a). The 17 chronic values for invertebrate species range from 2.83 (D. pulex) to

34.6 μg/L (C. dubia); and the 12 chronic values for the fish species range from <5 (brook trout) to

60.4 μg/L (northern pike). Of the 29 chronic tests, comparable acute values are available for 18 of

the tests (Table 2c). The relationship between acute toxicity values and ACRs is presented in Figure

6. The supporting acute and chronic test values for the ACRs and the species mean ACRs are
19

presented in Table 2c. For the 11 tests in Table 2a with chronic values both from a regression

EC20 and the geometric mean of the NOAEC and LOAEC, the EC20 averaged 81% of the

geometric mean, demonstrating the similar level of protection for the two approaches.

Overall, individual ACRs varied from <1 (0.55) for C. dubia (Oris et al., 1991) to 191.6 for

the snail, Campeloma decisum (Arthur and Leonard, 1970). Species mean acute-chronic ratios

ranged from 1.48 in saltwater for the sheepshead minnow (Hughes et al., 1989) to 171.2 in

freshwater for the snail, C. decisum. Pursuant to the Guidelines (Stephan et al., 1985),

consideration was given to calculating the FACR based on all ACRs within a factor of 10, but

because there appeared to be a relationship between acute sensitivity and ACRs (Figure 6), the

FACR was derived from data for species whose SMAVs were close to the FAV. The FACR of

3.22 was calculated as the geometric mean of the ACRs for sensitive freshwater species, C. dubia,

D. magna, D. pulex, O. tshawytscha, and O. mykiss along with the one saltwater ACR for C.

variegatus (Table 2b). Based on the normalization water chemistry conditions used for illustrative

purposes in the document, the freshwater site specific FAV value is 4.67 μg/L, which divided by the

FACR of 3.22 results in a freshwater FCV of 1.45 μg/L dissolved Cu.
20

5.0 PLANT DATA

Copper has been widely used as an algicide and herbicide for nuisance aquatic plants

(McKnight et al., 1983). Although copper is known as an inhibitor of photosynthesis and plant

growth, toxicity data on individual species suitable for deriving aquatic life criteria (Table 4) are not

numerous.

The relationship of copper toxicity to the complexing capacity of the water or the culture

medium is now widely recognized (Gächter et al., 1973; Petersen, 1982), and several studies have

used algae to “assay” the copper complexing capacity of both fresh and salt waters (Allen et al.,

1983; Lumsden and Florence, 1983; Rueter, 1983). It has also been shown that algae are capable of

excreting complexing substances in response to copper stress (McKnight and Morel, 1979; Swallow

et al., 1978; van den Berg et al., 1979). Foster (1982) and Stokes and Hutchinson (1976) have

identified resistant strains and/or species of algae from copper (or other metal) impacted

environments. A portion of this resistance probably results from induction of the chelate-excretion

mechanism. Chelate excretion by algae may also serve as a protective mechanism for other aquatic

organisms in eutrophic waters; that is, where algae are capable of maintaining free copper activities

below harmful concentrations.

Copper concentrations from 1 to 8,000 μg/L have been shown to inhibit growth of various

freshwater plant species. Very few of these tests, though, were accompanied by analysis of actual

copper exposure concentrations. Notable exceptions are freshwater tests with green alga including

Chlamydomonas reinhardtii (Schafer et al., 1993; Winner and Owen, 1991b), which is the only

flow-through, measured test with an aquatic plant, Chlorella vulgaris and Selenastrum

capricornutum (Blaylock et al., 1985). There is also a measured test with duckweed, Lemna minor

(Taraldsen and Norberg-King, 1990). 

A direct comparison between the freshwater plant data and the BLM derived criteria is

difficult to make without a better understanding of the composition of the algal media used for

different studies (e.g., DOC, hardness, and pH) because these factors influence the applicable

criteria comparison. BLM derived criteria for certain water conditions, such as low to mid-range

pH, hardness up to 100 mg/L as CaCO3, and low DOC are in the range of, if not lower than, the

lowest reported toxic endpoints for freshwater algal species and would therefore appear protective

of plant species. In other water quality conditions BLM-derived criteria may be significantly higher

(see Figure 5).

Two publications provide data for the red algae Champia parvula that indicate that

reproduction of this species is especially sensitive to copper. The methods manual (U.S. EPA 1988)

for whole effluent toxicity (WET) testing contains the results of six experiments showing nominal

reproduction LOECs from 48-hr exposures to 1.0 to 2.5 μg/L copper (mean 2.0 μg/L); these tests

used a mixture of 50 percent sterile seawater and 50 percent GP2 medium copper. The second

study by Morrison et al. (1989) evaluated interlaboratory variation of the 48-hr WET test

procedure; this six-test study gave growth EC50 values from 0.8 to 1.9 μg/L (mean 1.0 μg/L).

Thus, there are actually 12 tests that provide evidence of significant reproductive impairment in C.

parvula at nominal copper concentrations between 0.8 and 2.5 μg/L. For these studies though, the

dilution water source was not identified.
21

One difficulty in assessing these data is the uncertainty of the copper concentration in the test

solutions, primarily with respect to any background copper that might be found in the dilution

water, especially with solutions compounded from sea salts or reagents. Thus, with a CCC of 1.9

μg/L dissolved copper, the significance of a 1 or 2 μg/L background copper level to a 1 to 3 μg/L

nominal effect level can be considerable.

The reproduction of other macroalgae appears to be generally sensitive to copper, but not to

the extent of Champia. Many of these other macroalgae appear to have greater ecological

significance than Champia, several forming significant intertidal and subtidal habitats for other

saltwater organisms, as well as being a major food source for grazers. Reproductive and growth

effects on the other species of macroalgae sometimes appear to occur at copper concentrations

between 5 and 10 μg/L (Appendix B, Other Data). Thus, most major macrophyte groups seem to

be adequately protected by the CMC and CCC, but appear similar in sensitivity to some of the more

sensitive groups of saltwater animals.

6.0 OTHER DATA

Many of the data identified for this effort are listed in Appendix B, Other Data, for various

reasons, including exposure durations other than 96 hours with the same species reported in Table

1, and some exposures lasting up to 30 days. Acute values for test durations less than 96 hours are

available for several species not shown in Table 1. Still, these species have approximately the same

sensitivities to copper as species in the same families listed in Table 1. Reported LC50s at 200 hours

for chinook salmon and rainbow trout (Chapman, 1978) differ only slightly from 96-hour LC50s

reported for these same species in the same water. 

A number of other acute tests in Appendix B were conducted in dilution waters that were not

considered appropriate for criteria development. Brungs et al. (1976) and Geckler et al. (1976)

conducted tests with many species in stream water that contained a large amount of effluent from a

sewage treatment plant. Wallen et al. (1957) tested mosquitofish in a turbid pond water. Until

chemical measurements that correlate well with the toxicity of copper in a wide variety of waters

are identified and widely used, results of tests in unusual dilution waters, such as those in Appendix

B, will not be very useful for deriving water quality criteria.

Appendix B also includes tests based on physiological effects, such as changes in appetite,

blood parameters, stamina, etc. These were included in Appendix B because they could not be

directly interpreted for derivation of criteria. For the reasons stated in this section above, data in

Appendix B was not used for criteria derivation.

A direct comparison of a particular test result to a BLM-derived criterion is not always

straightforward, particularly if complete chemical characterization of the test water is not available.

Such is the case for a number of studies included in Appendix B. While there are some test results

with effect concentrations below the example criteria concentrations presented in this document,

these same effect concentrations could be above criteria derived for other normalization chemistries,

raising the question as to what is the appropriate comparison to make. For example, Appendix B

includes an EC50 for D. Pulex of 3.6 μg/L (Koivisto et al., 1992) at an approximate hardness of 25

mg/L (33 mg/L as CaCO3

). Yet, example criteria at a hardness of 25 mg/L (as CaCO3

) (including

those in Figure 6) range from 0.23 μg/L (DOC = 0.1 mg/L) to 4.09 μg/L (DOC = 2.3 mg/L) based
22

on the DOC concentration selected for the synthetic water recipe. The chemical composition for the

Koivisto et al. (1992) study would dictate what the appropriate BLM criteria comparison should be. 

Based on the expectation that many of the test results presented in Appendix B were

conducted in laboratory dilution water with low levels of DOC, the appropriate comparison would

be to the criteria derived from low DOC waters. Comparing many of the values in Appendix B to

the example criteria presented in this document, it appears that a large proportion of Appendix B

values are above these concentration levels. This is a broad generalization though and as stated

previously, all important water chemistry variables that affect toxicity of copper to aquatic

organisms should be considered before making these types of comparisons. 

Studies not considered suitable for criteria development were placed in Appendix G, Unused

Data.

7.0 NATIONAL CRITERIA STATEMENT

The available toxicity data, when evaluated using the procedures described in the “Guidelines

for Deriving Numerical National Water Quality Criteria for the Protection of Aquatic Organisms

and Their Uses” indicate that freshwater aquatic life should be protected if the 24-hour average and

four-day average concentrations do not respectively exceed the acute and chronic criteria

concentrations calculated by the Biotic Ligand Model.

A return interval of 3 years between exceedances of the criterion continues to be EPA's

general recommendation. However, the resilience of ecosystems and their ability to recover differ

greatly. Therefore, scientific derivation of alternative frequencies for exceeding criteria may be

appropriate.

8.0 IMPLEMENTATION

The use of water quality criteria in designing waste treatment facilities and appropriate effluent

limits involves the use of an appropriate wasteload allocation model. Although dynamic models are

preferred for application of these criteria, limited data or other factors may make their use

impractical, in which case one should rely on a steady-state model. EPA recommends the interim

use of 1B3 or 1Q10 for criterion maximum concentration stream design flow and 4B3 or 7Q10 for

the criterion continuous concentration design flow in steady-state models. These matters are

discussed in more detail in the Technical Support Document for Water Quality-Based Toxics

Control (U.S. EPA, 1991).

With regard to BLM-derived freshwater criteria, to develop a site-specific criterion for a

stream reach, one is faced with determining what single criterion is appropriate even though a BLM

criterion calculated for the event corresponding to the input water chemistry conditions will be

time-variable. This is not a new problem unique to the BLM—hardness-dependent metals criteria

are also time-variable values. Although the variability of hardness over time can be characterized,

EPA has not provided guidance on how to calculate site-specific criteria considering this variability.

Multiple input parameters for the BLM could complicate the calculation of site-specific criteria

because of their combined effects on variability. Another problem arise from potential scarcity of

data from small stream reaches with small dischargers. The EPA is currently exploring two
23

approaches to fill data gaps in such situations. One potential approach is the selection of values

based on geography, the second approach is based on correlations between measured parameters

and missing parameter measurements. A companion document in the form of Supplementary

Training Materials, addressing issues related to data requirements, implementation,

permitting, and monitoring will be released via EPA's website following the publication of

this criteria document.


Table 1. Acute Toxicity of Copper to Freshwater Animals

 Speciesa Organism Age, Size, or Lifestage Methodb Chemicalc Reported LC50 or EC50 (total μg/L)d Reported LC50 or EC50 (Diss. μg/L)e BLM Data Label BLM Normalized LC50 or EC50 (μg/L)f Species Mean 

Value (μg/

Worm, adult (mixed age) S,M,T N 130 --- LUVA01S 37.81 48.41 Lumbriculus variegat adult (mixed age) S,M,T N 270 --- LUVA02S 55.39 

adult (mixed age) S,M,T N 500 --- LUVA03S 54.18 

Snail, 1.1-2.7 cm F,M,T S 2000 --- CADE01F 4319 3573 

Campeloma 1.1-2.7 cm F,M,T S 1400 --- CADE02F 2956 

Snail,Juga plicifera

adult F,M,T C 15 --- JUPL01F 12.31 12.31 

Snail,Lithoglyphus virens

adult F,M,T C 8 --- LIVI01F 6.67 6.67 

Snail, 0.4-0.7 cm F,M,T S 41 --- PHIN01F 21.81 20.41 Physa integra 0.4-0.7 cm F,M,T S 37 --- PHIN02F 19.09 Freshwater mussel, juvenile S,M,T S 27 --- ACPE01S 10.36 11.33 Actinonaias juvenile S,M,T S <29 --- ACPE02S 12.39 Freshwater mussel, 1-2 d juv S,M,T S 86 --- UTIM01S 177.9 52.51 Utterbackia imbecillis 1-2 d juv S,M,T S 199 --- UTIM02S 172.3 

juvenile S,M,T N 76 --- UTIM03S 40.96 

juvenile S,M,T N 85 --- UTIM04S 43.22 juvenile S,M,T N 41 --- UTIM05S 24.12 juvenile S,M,T S 79 --- UTIM06S 39.04 juvenile S,M,T S 72 --- UTIM07S 39.96 juvenile S,M,T S 38 --- UTIM08S 28.31 

Cladoceran, <4 h S,M,T C 19 --- CEDU01S 10.28 5.93 Ceriodaphnia dubia <4 h S,M,T C 17 --- CEDU02S 9.19 

<12 h S,M,D --- - 25 CEDU03S 7.98 

<12 h S,M,D --- - 17 CEDU04S 5.25 

<12 h S,M,D --- - 30 CEDU05S 9.80 

<12 h S,M,D --- - 24 CEDU06S 7.63 

<12 h S,M,D --- - 28 CEDU07S 9.06 

<12 h S,M,D --- - 32 CEDU08S 10.56 

<12 h S,M,D --- - 23 CEDU09S 7.28 

<12 h S,M,D --- - 20 CEDU10S 6.25 

<12 h S,M,D --- - 19 CEDU11S 5.91 

<12 h S,M,D --- - 26 CEDU12S 3.10 

<12 h S,M,D --- - 21 CEDU13S 2.46 

<12 h S,M,D --- - 27 CEDU14S 3.24 

<12 h S,M,D --- - 37 CEDU15S 4.66 

<12 h S,M,D --- - 34 CEDU16S 4.22 

<12 h S,M,D --- - 67 CEDU17S 5.50 

<12 h S,M,D --- - 38 CEDU18S 2.72 

<12 h S,M,D --- - 78 CEDU19S 6.74 

<12 h S,M,D --- - 81 CEDU20S 7.10 

<12 h S,M,D --- - 28 CEDU21S 4.10 

24
Table 1. Acute Toxicity of Copper to Freshwater Animals

 Speciesa Organism Age, Size, or Lifestage Methodb Chemicalc Reported LC50 or EC50 (total μg/L)d Reported LC50 or EC50 (Diss. μg/L)e BLM Data Label BLM Normalized LC50 or EC50 (μg/L)f Species Mean 

Value (μg/

<12 h S,M,D --- - 84 CEDU22S 10.74 

<12 h S,M,T S 13.4 --- CEDU23S 6.19 <24 h R,M,T,D S 6.98 5.54 CEDU24R 5.03 

Cladoceran, 1 d S,M,T C 9.1 --- DAMA01S 3.42 6.00 Daphnia magna 1 d S,M,T C 11.7 --- DAMA02S 4.43 

<2 h S,M,T C 6.6 --- DAMA03S 2.50 <2 h S,M,T C 9.9 --- DAMA04S 3.78 

1 d S,M,T C 11.7 --- DAMA05S 13.46 

<4 h S,M,T C 6.7 --- DAMA06S 8.21 

1 d S,M,T C 9.1 --- DAMA07S 4.40 

<2 h S,M,T C 5.2 --- DAMA08S 2.16 

<24 h S,M,T S 41.2 --- DAMA09S 21.55 <24 h S,M,T S 10.5 --- DAMA10S 5.63 <24 h S,M,T S 20.6 --- DAMA11S 11.31 <24 h S,M,T S 17.3 --- DAMA12S 9.48 <24 h S,M,T S 70.7 --- DAMA13S 33.58 <24 h S,M,T S 31.3 --- DAMA14S 16.90 <24 h S,M,I S 7.1 --- DAMA15S 2.67 <24 h S,M,I S 16.4 --- DAMA16S 4.26 <24 h S,M,I S 39.9 --- DAMA17S 5.18 <24 h S,M,I S 18.7 --- DAMA18S 3.39 <24 h S,M,I S 18.9 --- DAMA19S 1.99 <24 h S,M,I S 39.7 --- DAMA20S 3.04 <24 h S,M,I S 46 --- DAMA21S 8.93 <24 h S,M,I S 71.9 --- DAMA22S 9.97 <24 h S,M,I S 57.2 --- DAMA23S 5.76 <24 h S,M,I S 67.8 --- DAMA24S 4.16 <24 h S,M,T C 26 --- DAMA25S 10.34 <24 h S,M,T C 30 --- DAMA26S 9.04 <24 h S,M,T C 38 --- DAMA27S 9.84 <24 h S,M,T C 69 --- DAMA28S 12.31 <24 h S,M,T,D S 4.8 --- DAMA29S 1.22 <24 h S,M,T,D S 7.4 --- DAMA30S 16.29 <24 h S,M,T,D S 6.5 --- DAMA31S 2.11 

Cladoceran, --- S,M,T S 11.4 --- DAPC01S 1.63 2.73 Daphnia pulicaria --- S,M,T S 9.06 --- DAPC02S 1.04 

--- S,M,T S 7.24 --- DAPC03S 0.88 --- S,M,T S 10.8 --- DAPC04S 1.13 --- S,M,T S 55.4 --- DAPC05S 8.81 --- S,M,T S 55.3 --- DAPC06S 6.03 --- S,M,T S 53.3 --- DAPC07S 4.12 --- S,M,T S 97.2 --- DAPC08S 3.94 

25
Table 1. Acute Toxicity of Copper to Freshwater Animals

 Speciesa Organism Age, Size, or Lifestage Methodb Chemicalc Reported LC50 or EC50 (total μg/L)d Reported LC50 or EC50 (Diss. μg/L)e BLM Data Label BLM Normalized LC50 or EC50 (μg/L)f Species Mean 

Value (μg/

--- S,M,T S 199 --- DAPC09S 3.01 --- S,M,T S 213 --- DAPC10S 7.63 --- S,M,T S 165 --- DAPC11S 5.78 --- S,M,T S 35.5 --- DAPC12S 1.83 --- S,M,T S 78.8 --- DAPC13S 2.36 --- S,M,T S 113 --- DAPC14S 1.06 --- S,M,T S 76.4 --- DAPC15S 2.36 --- S,M,T S 84.7 --- DAPC16S 6.62 --- S,M,T S 184 --- DAPC17S 7.14 --- S,M,T S 9.3 --- DAPC18S 1.11 --- S,M,T S 17.8 --- DAPC19S 2.11 --- S,M,T S 23.7 --- DAPC20S 2.67 --- S,M,T S 27.3 --- DAPC21S 2.77 --- S,M,T S 25.2 --- DAPC22S 2.81 --- S,M,T S 25.1 --- DAPC23S 2.60 --- S,M,T S 25.1 --- DAPC24S 2.31 

Cladoceran,Scapholeberis sp.

adult S,M,T C 18 --- SCSP01S 9.73 9.73 

Amphipod, 1-3 d F,M,T S 22 --- GAPS01F 10.39 9.60 Gammarus 1-3 d F,M,T S 19 --- GAPS02F 8.86 Amphipod, 7-14 d S,M,T N 17 --- HYAZ01S 12.19 12.07 

Hyalella azteca 7-14 d S,M,T N 24 --- HYAZ02S 9.96 

7-14 d S,M,T N 87 --- HYAZ03S 15.77 

<7 d S,M,T S 24.3 --- HYAZ04S 8.26 <7 d S,M,T S 23.8 --- HYAZ05S 8.09 <7 d S,M,T S 8.2 --- HYAZ06S 15.49 <7 d S,M,T S 10 --- HYAZ07S 18.80 

Stonefly,Acroneuria lycorias

--- S,M,T S 8300 --- ACLY01S 20636 20636 

Midge,Chironomus 

4th instar S,M,T S 739 --- CHDE01S 1987 1987 

Shovelnose sturgeon, Scaphirhynchus 

fry, 6.01 cm, 0.719 g S,M,T S 160 --- SCPL01S 69.63 69.63 

Apache trout,Oncorhynchus 

larval, 0.38 g S,M,T S 70 --- ONAP01S 32.54 32.54 

Lahontan cutthroat larval, 0.34 g S,M,T S 80 --- ONCL01S 34.26 32.97 Oncorhynchus clarki henshawi

larval, 0.57 g S,M,T S 60 --- ONCL02S 24.73 

26
Table 1. Acute Toxicity of Copper to Freshwater Animals

 Speciesa Organism Age, Size, or Lifestage Methodb Chemicalc Reported LC50 or EC50 (total μg/L)d Reported LC50 or EC50 (Diss. μg/L)e BLM Data Label BLM Normalized LC50 or EC50 (μg/L)f Species Mean 

Value (μg/

Cutthroat trout, 7.4 cm, 4.2 g F,M,T,D C 398.91 367 ONCL03F 67.30 Oncorhynchus clarki 6.9 cm, 3.2 g F,M,T,D C 197.87 186 ONCL04F 44.91 

8.8 cm, 9.7 g F,M,T,D C 41.35 36.8 ONCL05F 21.87 8.1 cm, 4.4 g F,M,T,D C 282.93 232 ONCL06F 51.94 6.8 cm, 2.7 g F,M,T,D C 186.21 162 ONCL07F 111.3 7.0 cm, 3.2 g F,M,T,D C 85.58 73.6 ONCL08F 39.53 8.5 cm, 5.2 g F,M,T,D C 116.67 91 ONCL09F 19.63 7.7 cm, 4.4 g F,M,T,D C 56.20 44.4 ONCL10F 18.81 8.9 cm, 5.7 g F,M,T,D C 21.22 15.7 ONCL11F 10.60 

Pink salmon, alevin (newly hatched) F,M,T S 143 --- ONGO01F 41.65 40.13 

Oncorhynchus gorbu alevin F,M,T S 87 --- ONGO02F 19.70 

fry F,M,T S 199 --- ONGO03F 78.76 

Coho salmon, 6 g R,M,T,I --- 164 --- ONKI01R 106.09 22.93 

Oncorhynchus kisutc parr F,M,T C 33 --- ONKI02F 20.94 

adult, 2.7 kg F,M,T C 46 --- ONKI03F 32.66 

fry F,M,T,D,I --- 61 49 ONKI04F 12.67 

smolt F,M,T,D,I --- 63 51 ONKI05F 13.19 

fry F,M,T,D,I --- 86 58 ONKI06F 11.95 

parr F,M,T,D,I --- 103 78 ONKI07F 22.98 

Rainbow trout, larval, 0.67 g S,M,T S 110 --- ONMY01S 41.64 22.19 Oncorhynchus mykis larval, 0.48 g S,M,T S 50 --- ONMY02S 25.26 

larval, 0.50 g S,M,T S 60 --- ONMY03S 29.46 

swim-up, 0.25 g R,M,T,D C 46.7 40 ONMY04R 10.90 swim-up, 0.25 g R,M,T,D C 24.2 19 ONMY05R 9.04 

swim-up, 0.20-0.24 g R,M,T,D C 0 3.4 ONMY06R 5.02 

swim-up, 0.20-0.24 g R,M,T,D C 0 8.1 ONMY07R 11.97 

swim-up, 0.20-0.24 g R,M,T,D C 0 17.2 ONMY08R 13.80 swim-up, 0.20-0.24 g R,M,T,D C 0 32 ONMY09R 23.84 

alevin F,M,T C 28 --- ONMY10F 20.30 

swim-up, 0.17 g F,M,T C 17 --- ONMY11F 12.54 

parr, 8.6 cm, 6.96 g F,M,T C 18 --- ONMY12F 9.87 

smolt, 18.8 cm, 68.19 g F,M,T C 29 --- ONMY13F 22.48 

1 g F,M,T,D C - 169 ONMY14F 23.41 

4.9 cm F,M,T,D C - 85.3 ONMY15F 10.20 

6.0 cm, 2.1 g F,M,T,D C - 83.3 ONMY16F 9.93 

6.1 cm, 2.5 g F,M,T,D C - 103 ONMY17F 12.71 

2.6 g F,M,T,D C - 274 ONMY18F 44.54 

4.3 g F,M,T,D C - 128 ONMY19F 16.51 

9.2 cm, 9.4 g F,M,T,D C - 221 ONMY20F 33.33 

9.9 cm, 11.5 g F,M,T,D C - 165 ONMY21F 22.70 

11.8 cm, 18.7 g F,M,T,D C - 197 ONMY22F 28.60 

13.5 cm, 24.9 g F,M,T,D C - 514 ONMY23F 99.97 

27
Table 1. Acute Toxicity of Copper to Freshwater Animals

 Speciesa Organism Age, Size, or Lifestage Methodb Chemicalc Reported LC50 or EC50 (total μg/L)d Reported LC50 or EC50 (Diss. μg/L)e BLM Data Label BLM Normalized LC50 or EC50 (μg/L)f Species Mean 

Value (μg/

13.4 cm, 25.6 g F,M,T,D C - 243 ONMY24F 37.88 

6.7 cm, 2.65 g F,M,T C 2.8 --- ONMY25F 7.00 

parr F,M,T,D,I --- 90 68 ONMY26F 19.73 

swim-up, 0.29 g F,M,T,D C 19.6 18 ONMY27F 8.10 swim-up, 0.25 g F,M,T,D C 12.9 12 ONMY28F 32.15 swim-up, 0.23 g F,M,T,D C 5.9 5.7 ONMY29F 24.80 swim-up, 0.23 g F,M,T,D C 37.8 35 ONMY30F 16.16 swim-up, 0.26 g F,M,T,D C 25.1 18 ONMY31F 37.66 swim-up, 0.23 g F,M,T,D C 17.2 17 ONMY32F 24.19 

0.64 g, 4.1 cm F,M,T,D C 101 --- ONMY33F 39.73 

0.35 g, 3.4 cm F,M,T,D C 308 --- ONMY34F 85.83 0.68 g, 4.2 cm F,M,T,D C 93 --- ONMY35F 95.9 0.43 g, 3.7 cm F,M,T,D C 35.9 --- ONMY36F 50.83 0.29 g, 3.4 cm F,M,T,D C 54.4 --- ONMY37F 47.69 

Sockeye salmon, alevin (newly hatched) F,M,T S 190 --- ONNE01F 71.73 54.82 

Oncorhynchus nerka alevin F,M,T S 200 --- ONNE02F 79.52 

alevin F,M,T S 100 --- ONNE03F 23.74 alevin F,M,T S 110 --- ONNE04F 27.22 alevin F,M,T S 130 --- ONNE05F 35.36 

fry F,M,T S 150 --- ONNE06F 45.37 

smolt, 5.5 g F,M,T S 210 --- ONNE07F 87.77 

smolt, 5.5 g F,M,T S 170 --- ONNE08F 57.53 smolt, 5.5 g F,M,T S 190 --- ONNE09F 71.73 smolt, 4,8 g F,M,T S 240 --- ONNE10F 114.4 

Chinook salmon, alevin, 0.05 g F,M,T C 26 --- ONTS01F 14.48 25.02 Oncorhynchus tshaw swim-up, 0.23 g F,M,T C 19 --- ONTS02F 10.44 

parr, 9.6 cm, 11.58 g F,M,T C 38 --- ONTS03F 28.30 

smolt, 14.4 cm, 32.46 g F,M,T C 26 --- ONTS04F 20.09 

3 mo, 1.35 g F,M,T,I C 10.2 --- ONTS05F 19.41 

3 mo, 1.35 g F,M,T,I C 24.1 --- ONTS06F 30.91 

3 mo, 1.35 g F,M,T,I C 82.5 --- ONTS07F 32.74 

3 mo, 1.35 g F,M,T,I C 128.4 --- ONTS08F 20.66 

swim-up, 0.36-0.45 g F,M,T,D C 0 7.4 ONTS09F 36.49 

swim-up, 0.36-0.45 g F,M,T,D C 0 12.5 ONTS10F 30.85 

swim-up, 0.36-0.45 g F,M,T,D C 0 14.3 ONTS11F 31.49 

swim-up, 0.36-0.45 g F,M,T,D C 0 18.3 ONTS12F 48.56 

28
Table 1. Acute Toxicity of Copper to Freshwater Animals

 Speciesa Organism Age, Size, or Lifestage Methodb Chemicalc Reported LC50 or EC50 (total μg/L)d Reported LC50 or EC50 (Diss. μg/L)e BLM Data Label BLM Normalized LC50 or EC50 (μg/L)f Species Mean 

Value (μg/

Bull trout, 0.130 g, 2.6 cm F,M,T,D C 228 --- SACO01F 69.70 68.31 Salvelinus confluentu 0.555 g, 4.0 cm F,M,T,D C 207 --- SACO02F 63.62 

0.774 g, 4.5 cm F,M,T,D C 66.6 --- SACO03F 74.18 1.520 g, 5.6 cm F,M,T,D C 50 --- SACO04F 63.60 1.160 g, 5.2 cm F,M,T,D C 89 --- SACO05F 71.11 

Chiselmouth,Acrocheilus 

4.6 cm, 1.25 g F,M,T C 143 --- ACAL01F 216.3 216.3 

Bonytail chub,Gila elegans

larval, 0.29 g S,M,T S 200 --- GIEL01S 63.22 63.22 

Golden shiner,Notemigonus crysoleucas

--- F,M,T C 84600 --- NOCR01F 107860 107860 

Fathead minnow, adult, 40 mm S,M,T S 310 --- PIPR01S 266.3 69.63 Pimephales promela adult, 40 mm S,M,T S 120 --- PIPR02S 105.61 

adult, 40 mm S,M,T S 390 --- PIPR03S 207.3 

--- S,M,T C 55 --- PIPR04S 38.08 --- S,M,T C 85 --- PIPR05S 70.71 

<24 h S,M,T N 15 --- PIPR06S 11.23 

<24 h S,M,T N 44 --- PIPR07S 18.03 <24 h S,M,T N >200 --- PIPR08S 24.38 

<24 h, 0.68 mg S,M,T S 4.82 --- PIPR09S 8.87 <24 h, 0.68 mg S,M,T S 8.2 --- PIPR10S 16.72 <24 h, 0.68 mg S,M,T S 31.57 --- PIPR11S 25.15 <24 h, 0.68 mg S,M,T S 21.06 --- PIPR12S 17.67 <24 h, 0.68 mg S,M,T S 35.97 --- PIPR13S 21.24 <24 h, 0.68 mg S,M,T S 59.83 --- PIPR14S 16.64 <24 h, 0.68 mg S,M,T S 4.83 --- PIPR15S 5.92 <24 h, 0.68 mg S,M,T S 70.28 --- PIPR16S 13.34 <24 h, 0.68 mg S,M,T S 83.59 --- PIPR17S 8.22 <24 h, 0.68 mg S,M,T S 182 --- PIPR18S 13.91 

larval, 0.32 g S,M,T S 290 --- PIPR19S 73.92 

larval, 0.56 g S,M,T S 630 --- PIPR20S 157.9 

larval, 0.45 g S,M,T S 400 --- PIPR21S 103.2 

larval, 0.39 g S,M,T S 390 --- PIPR22S 161.7 

3.2-5.5 cm, 0.42-3.23 g S,M,T S 450 --- PIPR23S 152.9 

2.8-5.1 cm, 0.30-2.38 g S,M,T S 297 --- PIPR24S 77.75 1.9-4.6 cm, 0.13-1.55 g S,M,T S 311 --- PIPR25S 67.56 3.0-4.8 cm, 0.23-1.36 g S,M,T S 513 --- PIPR26S 76.36 

<24 h S,M,T,D S 62.23 53.96 PIPR27S 25.70 

<24 h S,M,T,D S 190.5 165.18 PIPR28S 87.89 

<24 h S,M,T,D S 68.58 59.46 PIPR29S 28.59 

<24 h S,M,T,D S 168.91 146.46 PIPR30S 89.18 

29
Table 1. Acute Toxicity of Copper to Freshwater Animals

 Speciesa Organism Age, Size, or Lifestage Methodb Chemicalc Reported LC50 or EC50 (total μg/L)d Reported LC50 or EC50 (Diss. μg/L)e BLM Data Label BLM Normalized LC50 or EC50 (μg/L)f Species Mean 

Value (μg/

<24 h S,M,T,D S 94.62 82.04 PIPR31S 49.27 

<24 h S,M,T,D S 143.51 124.43 PIPR32S 104.90 <24 h S,M,T,D S 120.65 103.76 PIPR33S 86.54 <24 h S,M,T,D S 196.85 167.32 PIPR34S 122.0 <24 h S,M,T,D S 133.35 120.02 PIPR35S 75.0 <24 h S,M,T,D S 184.15 169.42 PIPR36S 122.2 <24 h S,M,T,D S 304.8 268.22 PIPR37S 78.5 <24 h S,M,T,D S 292.1 242.44 PIPR38S 201.5 <24 h S,M,T,D S 133.35 113.35 PIPR39S 100.75 <24 h S,M,T,D S 92.71 77.88 PIPR40S 72.95 <24 h S,M,T,D S 152.4 128.02 PIPR41S 112.9 <24 h S,M,T,D S 177.8 151.13 PIPR42S 136.3 <24 h S,M,T,D S 203.2 166.62 PIPR43S 136.0 <24 h S,M,T,D S 190.5 163.83 PIPR44S 147.7 <24 h S,M,T,D S 196.85 157.48 PIPR45S 125.9 <24 h S,M,T,D S 234.95 199.71 PIPR46S 157.4 <24 h S,M,T,D S 146.05 128.52 PIPR47S 127.8 <24 h S,M,T,D S 171.45 150.88 PIPR48S 153.9 <24 h S,M,T,D S 152.4 131.06 PIPR49S 114.57 <24 h S,M,T,D S 184.15 160.21 PIPR50S 131.3 <24 h S,M,T,D S 203.2 182.88 PIPR51S 130.9 <24 h S,M,T,D S 203.2 180.85 PIPR52S 105.76 <24 h S,M,T,D S 203.2 176.78 PIPR53S 128.8 <24 h S,M,T,D S 222.25 188.91 PIPR54S 122.1 <24 h S,M,T,D S 146.05 125.60 PIPR55S 111.87 <24 h S,M,T,D S 139.7 117.35 PIPR56S 85.45 <24 h S,M,T,D S 139.7 114.55 PIPR57S 83.10 <24 h S,M,T,D S 152.4 126.49 PIPR58S 85.82 <24 h S,M,T,D S 203.2 172.72 PIPR59S 110.0 <24 h S,M,T,D S 196.85 167.32 PIPR60S 106.46 <24 h S,M,T,D S 266.7 226.70 PIPR61S 133.4 <24 h S,M,T,D S 99.06 84.20 PIPR62S 138.0 <24 h S,M,T,D S 111.13 97.79 PIPR63S 165.8 <24 h S,M,T,D S 78.74 70.08 PIPR64S 114.8 <24 h S,M,T,D S 92.71 81.58 PIPR65S 121.5 <24 h S,M,T,D S 85.09 77.43 PIPR66S 106.69 <24 h S,M,T,D S 123.19 110.87 PIPR67S 124.7 <24 h S,M,T,D S 165.1 151.89 PIPR68S 114.24 <24 h S,M,T,D S 190.5 175.26 PIPR69S 89.93 <24 h S,M,T,D S 165.1 145.29 PIPR70S 140.2 <24 h S,M,T,D S 127 111.76 PIPR71S 100.16 <24 h S,M,T,D S 92.08 79.18 PIPR72S 58.74 

30
Table 1. Acute Toxicity of Copper to Freshwater Animals

 Speciesa Organism Age, Size, or Lifestage Methodb Chemicalc Reported LC50 or EC50 (total μg/L)d Reported LC50 or EC50 (Diss. μg/L)e BLM Data Label BLM Normalized LC50 or EC50 (μg/L)f Species Mean 

Value (μg/

<24 h S,M,T,D S 66.68 60.01 PIPR73S 37.67 

<24 h S,M,T,D S 393.70 370.08 PIPR74S 163.3 <24 h S,M,T,D S 317.50 292.10 PIPR75S 252.2 <24 h S,M,T,D S 107.95 101.47 PIPR76S 169.6 <24 h S,M,T,D S 67.95 62.51 PIPR77S 146.5 <24 h S,M,T,D S 45.72 42.06 PIPR78S 126.3 <24 h S,M,T,D S 177.80 172.47 PIPR79S 197.6 <24 h S,M,T,D S 13.97 12.43 PIPR80S 28.13 <24 h S,M,T,D S 304.80 271.27 PIPR81S 149.2 <24 h S,M,T,D S 71.12 71.12 PIPR82S 105.76 <24 h S,M,T,D S 83.82 79.63 PIPR83S 108.41 <24 h S,M,T,D S 104.78 99.54 PIPR84S 114.7 <24 h S,M,T,D S 139.70 132.72 PIPR85S 137.8 <24 h S,M,T,D S 152.40 137.16 PIPR86S 114.8 <24 h S,M,T,D S 260.35 182.25 PIPR87S 114.8 <24 h S,M,T,D S 488.95 268.92 PIPR88S 122.1 <24 h S,M,T,D S 203.20 188.98 PIPR89S 147.5 <24 h S,M,T,D S 704.85 662.56 PIPR90S 185.0 <24 h S,M,T,D S 952.50 904.88 PIPR91S 197.1 <24 h S,M,T,D S 1244.60 995.68 PIPR92S 188.3 <24 h S,M,T,D S 1485.90 891.54 PIPR93S 135.5 <24 h S,M,T,D S 781.05 757.62 PIPR94S 181.4 <24 h S,M,T,D S 476.25 404.81 PIPR95S 172.5 <24 h S,M,T,D S 273.05 262.13 PIPR96S 191.4 <24 h S,M,T,D S 22.23 20.45 PIPR97S 59.14 <24 h S,M,T,D S 24.13 23.16 PIPR98S 64.08 <24 h S,M,T,D S 36.83 34.99 PIPR99S 97.49 <24 h S,M,T,D S 27.94 27.94 PIPR100S 78.99 <24 h S,M,T,D S 26.67 26.67 PIPR101S 72.86 <24 h S,M,T,D S 20.32 20.32 PIPR102S 50.73 <24 h S,M,T,D S 26.67 26.67 PIPR103S 68.24 <24 h S,M,T,D S 190.50 182.88 PIPR104S 146.6 <24 h S,M,T,D S 109.86 96.67 PIPR105S 93.76 <24 h S,M,T,D S 203.20 182.88 PIPR106S 128.86 <24 h S,M,T,D S 209.55 190.69 PIPR107S 113.0 <24 h S,M,T,D S 146.05 127.06 PIPR108S 101.01 <24 h S,M,T,D S 165.10 148.59 PIPR109S 120.9 <24 h S,M,T,D S 254.00 223.52 PIPR110S 137.6 <24 h S,M,T,D S 311.15 283.15 PIPR111S 142.9 <24 h S,M,T,D S 165.10 150.24 PIPR112S 106.74 <24 h S,M,T,D S 920.75 644.53 PIPR113S 131.9 <24 h S,M,T,D S 1073.15 697.55 PIPR114S 116.5 

31
Table 1. Acute Toxicity of Copper to Freshwater Animals

 Speciesa Organism Age, Size, or Lifestage Methodb Chemicalc Reported LC50 or EC50 (total μg/L)d Reported LC50 or EC50 (Diss. μg/L)e BLM Data Label BLM Normalized LC50 or EC50 (μg/L)f Species Mean 

Value (μg/

<24 h S,M,T,D S 1003.30 752.48 PIPR115S 109.8 <24 h S,M,T,D S 933.45 653.42 PIPR116S 123.2 <24 h S,M,T,D S 742.95 646.37 PIPR117S 129.6 <24 h S,M,T,D S 1879.60 939.80 PIPR118S 124.8 <24 h S,M,T,D S 266.70 253.37 PIPR119S 176.1 

--- F,M,T S 114.00 --- PIPR120F 17.99 --- F,M,T S 121.00 --- PIPR121F 19.70 --- F,M,T S 88.50 --- PIPR122F 13.27 --- F,M,T S 436.00 --- PIPR123F 78.50 --- F,M,T S 516.00 --- PIPR124F 50.09 --- F,M,T S 1586.00 --- PIPR125F 66.49 --- F,M,T S 1129.00 --- PIPR126F 73.03 --- F,M,T S 550.00 --- PIPR127F 42.76 --- F,M,T S 1001.00 --- PIPR128F 34.39 

30 d, 0.15 g F,M,T,D N 96.00 88.32 PIPR129F 39.58 

<24 h F,M,T,D S 31.75 27.94 PIPR130F 8.69 <24 h F,M,T,D S 117.48 105.73 PIPR131F 37.88 <24 h F,M,T,D S 48.26 40.06 PIPR132F 10.80 <24 h F,M,T,D S 73.03 64.26 PIPR133F 22.19 <24 h F,M,T,D S 59.06 49.02 PIPR134F 20.32 <24 h F,M,T,D S 78.74 67.72 PIPR135F 18.51 <24 h F,M,T,D S 22.23 18.67 PIPR136F 13.61 <24 h F,M,T,D S 6.99 6.15 PIPR137F 10.94 <24 h F,M,T,D S 22.23 20.45 PIPR138F 17.70 <24 h F,M,T,D S 107.32 93.36 PIPR139F 67.09 <24 h F,M,T,D S 292.10 245.36 PIPR140F 17.75 <24 h F,M,T,D S 81.28 72.34 PIPR141F 41.16 <24 h F,M,T,D S 298.45 229.81 PIPR142F 16.18 <24 h F,M,T,D S 241.30 195.45 PIPR143F 24.40 <24 h F,M,T,D S 133.35 109.35 PIPR144F 21.07 <24 h F,M,T,D S 93.98 78.00 PIPR145F 50.83 <24 h F,M,T,D S 67.95 45.52 PIPR146F 23.18 <24 h F,M,T,D S 4.76 4.38 PIPR147F 40.09 <24 h F,M,T,D S 13.97 12.43 PIPR148F 45.37 <24 h F,M,T,D S 29.85 26.86 PIPR149F 59.43 <24 h F,M,T,D S 59.69 51.33 PIPR150F 58.84 

Northern squawfish, larval, 0.32 g S,M,T S 380 --- PTLU01S 88.44 132.2 Ptychocheilus orego larval, 0.34 g S,M,T S 480 --- PTLU02S 197.6 

32
Table 1. Acute Toxicity of Copper to Freshwater Animals

 Speciesa Organism Age, Size, or Lifestage Methodb Chemicalc Reported LC50 or EC50 (total μg/L)d Reported LC50 or EC50 (Diss. μg/L)e BLM Data Label BLM Normalized LC50 or EC50 (μg/L)f Species Mean 

Value (μg/

Northern squawfish, 5.0 cm, 1.33 g F,M,T C 23 --- PTOR01F 17.02 14.61 Ptychocheilus orego 7.2 cm, 3.69 g F,M,T C 18 --- PTOR02F 12.54 Razorback sucker, larval, 0.31 g S,M,T S 220 --- XYTE01S 63.78 78.66 Xyrauchen texanus larval, 0.32 g S,M,T S 340 --- XYTE02S 97.0 Gila topminnow,Poeciliposis 

id t li

2.72 cm, 0.219 g S,M,T S 160 --- POAC01S 56.15 56.15 

Bluegill, 3.58 cm, 0.63 g R,M,D C - 2200 LEMA01R 2202 2231 

Lepomis macrochiru 12 cm, 35 g F,M,T S 1100 --- LEMA02F 2305 

2.8-6.8 cm F,M,T C 1000 --- LEMA03F 4200 

3.58 cm, 0.63 g F,M,D C - 1300 LEMA04F 1163 

Fantail darter, 3.7 cm S,M,T S 330 --- ETFL01S 117.7 124.3 Etheostoma flabellar 3.7 cm S,M,T S 341 --- ETFL02S 121.1 

3.7 cm S,M,T S 373 --- ETFL03S 122.8 3.7 cm S,M,T S 392 --- ETFL04S 136.6 

Greenthroat darter,Etheostoma 

2.26 cm, 0.133 g S,M,T S 260 --- ETLE01S 82.80 82.80 

Johnny darter, 3.9 cm S,M,T S 493 --- ETNI01S 167.3 178.3 Etheostoma nigrum 3.9 cm S,M,T S 483 --- ETNI02S 164.2 

3.9 cm S,M,T S 602 --- ETNI03S 200.1 3.9 cm S,M,T S 548 --- ETNI04S 183.9 

Fountain darter,Etheostoma rubrum

2.02 cm, 0.062 g S,M,T S 60 --- ETRU01S 22.74 22.74 

Boreal toad,Bufo boreas

tadpole, 0.012 g S,M,T S 120 --- BUBO01S 47.49 47.49 

a Species appear in order taxonomically, with invertebrates listed first, fish, and an amphibian listed last. Species within each genus are ordered alphabetically. Within each spec

test method (static, renewal, flow-through) and date.

b S = static, R = renewal, F = flow-through, U = unmeasured, M = measured, T = exposure concentrations were measured as total copper, D = exposure concentrations were mea

dissolved copper.

c S = copper sulfate, N = copper nitrate, C = copper chloride.d Values in this column are total copper LC50 or EC50 values as reported by the author.e Values in this column are dissolved copper LC50 or EC50 values either reported by the author or if the author did not report a dissolved value then a conversion factor (CF) was 

to the total copper LC50 to estimate dissolved copper values.g Underlined LC50s or EC50s not used to derive SMAV because considered extreme value.

33
Table 2a. Chronic Toxicity of Copper to Freshwater Animals

Chronic 

Valueb 

(μg/L)

EC20b 

(μg/L) 

Rotifer,Brachionus calyciflorus

LC,T Copper sulfate Intrinsic growth 

rate

85 2.5-5.0 3.54 - 3.54 3.54 

Snail,Campeloma decisum (Test 1)

LC,T Copper sulfate Survival 35-55 8-14.8 10.88 8.73 9.77 9.77 

Snail,Campeloma decisum (Test 2)

LC,T Copper sulfate Survival 35-55 8-14.8 10.88 10.94 

Cladoceran,Ceriodaphnia dubia (New River)

LC,D - Reproduction 179 6.3-9.9 7.90c 

(8.23)

- 19.3 19.3 

Cladoceran,Ceriodaphnia dubia (Cinch River)

LC,D - Reproduction 94.1 <19.3-19.3 <19.3 19.36c 

(20.17)

Cladoceran,Ceriodaphnia dubia

LC,T Copper sulfate Survival and 

reproduction

57 - 24.50 - 

Cladoceran,Ceriodaphnia dubia

LC,T Copper sulfate Survival and 

reproduction

57 - 34.60 - 

Cladoceran,Ceriodaphnia dubia

LC,T,D Copper chloride Reproduction 12-32 19.59 9.17 

Cladoceran,Daphnia magna

LC,T Copper chloride Reproduction 85 10-30 17.32 - 14.1 8.96 

Cladoceran,Daphnia magna

LC,T Copper chloride Carapace length 225 12.6-36.8 21.50 - 

Cladoceran,Daphnia magna

LC,T Copper chloride Reproduction 51 11.4-16.3 13.63 12.58 

Cladoceran,Daphnia magna

LC,T Copper chloride Reproduction 104 20-43 29.33 19.89 

Cladoceran,Daphnia magna

LC,T Copper chloride Reproduction 211 7.2-12.6 9.53 6.06 

Cladoceran,Daphnia pulex

LC,T Copper sulfate Survival 57.5 (No HA) 4.0-6.0 4.90 2.83 5.68 

Cladoceran,Daphnia pulex

LC,T Copper sulfate Survival 115 (No HA) 5.0-10.0 7.07 

Cladoceran,Daphnia pulex

LC,T Copper sulfate Survival 230 (0.15 HA) 10-15 12.25 9.16 

Hardness 

(mg/L as 

CaCO3)

Chronic 

Limits (μg/L) Species Testa Chemical Endpoint

Genus Mea

Chronic Val

(Total μg/L

Species Mean 

Chronic Value 

(Total μg/L)

Chronic Values

34
Table 2a. Chronic Toxicity of Copper to Freshwater Animals

Chronic 

Valueb 

(μg/L)

EC20b 

(μg/L) 

Hardness 

(mg/L as 

CaCO3)

Chronic 

Limits (μg/L) Species Testa Chemical Endpoint

Genus Mea

Chronic Val

(Total μg/L

Species Mean 

Chronic Value 

(Total μg/L)

Chronic Values

Caddisfly,Clistoronia magnifica

LC,T Copper chloride Emergence (adult 

1st gen)

26 8.3-13 10.39 7.67 7.67 7.67 

Rainbow trout,Oncorhynchus mykiss

ELS,T 

continuous

Copper chloride Biomass 120 27.77 23.8 11.9 

Rainbow trout,Oncorhynchus mykiss

ELS,T Copper sulfate Biomass 160-180 12-22 16.25 20.32 

Chinook salmon,Oncorhynchus tshawytscha

ELS,T Copper chloride Biomass 20-45 <7.4 <7.4 5.92 5.92 

Brown trout,Salmo trutta

ELS,T Copper sulfate Biomass 45.4 20.8-43.8 29.91 - 29.9 29.9 

Brook trout,Salvelinus fontinalis

PLC,T Copper sulfate Biomass 35.0 <5 -5 <5 - 12.5 19.7 

Brook trout,Salvelinus fontinalis

ELS,T Copper sulfate Biomass 45.4 22.3-43.5 31.15 - 

Lake trout,Salvelinus namaycush

ELS, T Copper sulfate Biomass 45.4 22.0-43.5 30.94 - 30.9 

Northern pike,Esox lucius

ELS, T Copper sulfate Biomass 45.4 34.9-104.4 60.36 - 60.4 60.4 

Bluntnose minnowPimephales notatus

LC,T Copper sulfate Egg production 172-230 <18-18 18.00 - 18.0 13.0 

Fathead minnow,Pimephales promelas

ELS,T,D - Biomass 45 9.38 9.38 

White sucker,Catostomus commersoni

ELS, T Copper sulfate Biomass 45.4 12.9-33.8 20.88 - 20.9 20.9 

Bluegill (larval),Lepomis macrochirus

ELS,T,D Copper sulfate Survival 44-50 21-40 28.98 27.15 27.2 27.2 

a LC = life-cycle; PLC = partial life-cyle; ELS = early life state; T = total copper; D = dissolved copper.b Results are based on copper, not the chemical.c Chronic values based on dissolved copper concentration. 

35
Table 2b. Chronic Toxicity of Copper to Saltwater Animals

Species Test Chemical Salinity (g/kg) Limits (μg/L) Chronic Value (μg/L) Chronic Value Dissolved (μg/L) ACR 

Sheepshead minnow,Cyprinodon variegatus ELS Copper chloride 30 172-362 249 206.7 1.48 

36
Table 2c. Acute-Chronic Ratios

 Species Hardness (mg/L 

as CaCO3)

Acute Value 

(μg/L)

Chronic 

Value (μg/L) Ratio Reference

Overall 

Ratio for 

Species

Snail, 35-55 1673a 8.73 191.61 Arthur and Leonard 1970

Campeloma decisum 35-55 1673a 10.94 152.95 Arthur and Leonard 1970 171.19

Cladoceran, 179 28.42b 7.90 3.60 Belanger et al. 1989

Ceriodaphnia dubia 94.1 63.33b 19.36 3.27 Belanger et al. 1989

57 13.4 24.5 0.55 Oris et al. 1991

-- 17.974c 9.17 1.96 Carlson et al. 1986 2.85g >

Cladoceran, 51 26 12.58 2.07 Chapman et al. Manuscript

Daphnia magna 104 33.76d 19.89 1.70 Chapman et al. Manuscript

211 69 6.06 11.39 Chapman et al. Manuscript 3.42 >

Cladoceran, 57.5 25.737 2.83 9.10 Winner 1985

Daphnia pulex 115 27.6 7.07 3.90 Winner 1985

230 28.79 9.16 3.14 Winner 1985 4.82 >

Rainbow trout,

Oncorhynchus mykiss 120 80 27.77 2.88 Seim et al. 1984 2.88 >

Chinook salmon,

Oncorhynchus tshawytscha 20-45 33.1 5.92 5.59 Chapman 1975, 1982 5.59 >

Bluntnose minnow,

Pimephales notatus 172-230 231.9e 18 12.88 Horning and Neiheisel 1979 12.88

Fathead minnow,

Pimephales promelas 45 106.875f 9.38 11.40 Lind et al. 1978 11.40

Bluegill, 

Lepomis macrochirus 21-40 1100 27.15 40.52 Benoit 1975 40.49

Sheepshead minnow,

Cyprinodon variegatus - 368 249 1.48 Hughes et al. 1989 1.48 > aGeometric mean of two values from Arthur and Leonard (1970) in Table 1.

 bGeometric mean of five values from Belanger et al. (1989) in Table 1. ACR is based on dissolved metal measurements.

 cGeometric mean of two values from Carlson et al. (1986) in Table 1.

 dGeometric mean of two values from Chapman manuscript in Table 1.

 eGeometric mean of two values of three values from Horning and Neiheisel (1979) in Appendix C.

 fGeometric mean of three values from Lind et al. (1978) in Table 1.

 gACR from Oris et al. (1991) not used in calculating overall ratio for species because it is <1.

FACR

Freshwater final acute-chronic ratio = 3.22

Saltwater final acute-chronic ratio = 3.22

37
Table 3a. Ranked Freshwater Genus Mean Acute Values with Species Mean

 Acute-Chronic Ratios

Rank GMAV Species SMAV (μg/L) ACR

27 107,860 Golden shiner, Notemigonus crysoleucas 107,860

26 20,636 Stonefly, Acroneuria lycorias 20,636

25 3,573 Snail, Campeloma decisum 3,573 171.19

24 2,231 Bluegill sunfish, Lepomis macrochirus 2,231 40.49

23 1,987 Midge, Chironomus decorus 1,987

22 216.3 Chiselmouth, Acrocheilus alutaceus 216.3

21 80.38 Fantail darter, Etheostoma flabellare 124.3

Greenthroat darter, Etheostoma lepidum 82.80

Johnny darter, Etheostoma nigrum 178.3

Fountain darter, Etheostoma rubrum 22.74

20 78.66 Razorback sucker, Xyrauchen texanus 78.66

19 69.63 Fathead minnow, Pimephales promelas 69.63 11.40

18 69.63 Shovelnose sturgeon, Scaphirhynchus platorynchus 69.63

17 68.31 Bull trout, Salvelinus confluentus 68.31

16 63.22 Bonytail chub, Gila elegans 63.22

15 56.15 Gila topminnow, Poeciliposis occidentalis 56.15

14 52.51 Freshwater mussel, Utterbackia imbecillis 52.51

13 48.41 Worm, Lumbriculus variegatus 48.41

12 47.49 Boreal toad, Bufo boreas 47.49

11 43.94 Colorado squawfish, Ptychocheilus lucius 132.2

Northern squawfish, Ptychocheilus oregonensis 14.61

10 31.39 Apache trout, Oncorhynchus apache 32.54

Cutthroat trout, Oncorhynchus clarki 32.97

Pink salmon, Oncorhynchus gorbuscha 40.13

Coho salmon, Oncorhynchus kisutch 22.93

Rainbow trout, Oncorhynchus mykiss 22.19 2.88

Sockeye salmon, Oncorhynchus nerka 54.82

Chinook salmon, Oncorhynchus tshawytscha 25.02 5.59

9 20.41 Snail, Physa integra 20.41

8 12.31 Snail, Juga plicifera 12.31

7 12.07 Amphipod, Hyalella azteca 12.07

6 11.33 Freshwater mussel, Actinonaias pectorosa 11.33

5 9.73 Cladoceran, Scapholeberis sp. 9.73

4 9.60 Amphipod, Gammarus pseudolimnaeus 9.60

3 6.67 Snail, Lithoglyphus virens 6.67

2 5.93 Cladoceran, Ceriodaphnia dubia 5.93 2.85

1 4.05 Cladoceran, Daphnia magna 6.00 3.42

Cladoceran, Daphnia pulicaria 2.73

38
Table 3b. Freshwater Final Acute Value (FAV) and Criteria Calculations

Calculated Freshwater FAV based on 4 lowest values: Total Number of GMAVs in Data Set = 27

Rank GMAV lnGMAV (lnGMAV)2

P = R/(n+1) SQRT(P)

4 9.600 2.261 5.114 0.143 0.378

3 6.670 1.897 3.599 0.107 0.327

2 5.930 1.780 3.170 0.071 0.267

1 4.050 1.398 1.954 0.036 0.189

Sum: 7.33671 13.83657 0.35714 1.16153

S = 4.374

L = 0.5641

A = 1.542

Calculated FAV = 4.674452

Calculated CMC = 2.337

Dissolved Copper Criterion Maximum Concentration (CMC) = 2.337 μg/L (for example normalization chemistry see Table 1, footnote f)

Criteria Lethal Accumulation (LA50) based on example normalization chemistry = 0.03395 nmol/g wet wt

Criterion Continuous Concentration (CCC) = 4.67445/3.22 = 1.4516932 μg/L (for example normalization chemistry see Table 1, footnote f)

S = Scale parameter or slope

L = Location parameter or intercept

P = Cumulative probability

A = lnFAV

39
Table 4. Toxicity of Copper to Freshwater Plants

Species Methoda Chemical Hardness (mg/L as CaCO3) Duration Effect Resultb (Total μg/L) 

Blue-green alga,Anabaena flos-aqua S,U Copper sulfate 65.2 96 hr EC75 (cell density) 200 YounBllue-green alga,Anabaena variabilis S,U Copper sulfate 65.2 - EC85 (wet weight) 100 YounBlue-green alga,Anabaena strain 7120 - - - - Lag in growth 64 LaubBlue-green alga,Chroococcus paris S,U Copper nitrate 54.7 10 days Growth reduction 100 Les aBlue-green alga,Microcystis aeruginosa S,U Copper sulfate 54.9 8 days Incipient inhibition 30 Bring

1976

Alga,Ankistrodesmus braunii - - - - Growth reduction 640 LaubGreen alga,Chlamydomonas sp. S,U Copper sulfate 68 10 days Growth inhibition 8,000 CairnGreen alga,Chlamydomonas reinhardtii S,M,T - 90 - 133 72 hr NOEC (deflagellation) 12.2-49.1 WinnGreen alga,Chlamydomonas reinhardtii S,M,T - 90 - 133 72 hr NOEC (cell density) 12.2-43.0 WinnGreen alga,Chlamydomonas reinhardtii F,M,T - 24 10 days EC50 (cell density) 31.5 Scha

Green alga,Chlorella pyrenoidosa S,U - - 96 hr ca. 12 hr lag in growth 1 Stee

1970

Green alga,Chlorella pyrenoidosa S,U - 54.7 - Growth inhibition 100 Stee

1970

Green alga,Chlorella pyrenoidosa S,U Copper sulfate 365 14 days EC50 (dry weight) 78-100 Bedn

Green alga,Chlorella pyrenoidosa S,U Copper sulfate 36.5 14 days EC50 (dry weight) 78-100 BednGreen alga,Chlorella pyrenoidosa S,U Copper sulfate 3.65 14 days EC50 (dry weight) 78-100 Bedn

1983

Green alga,Chlorella saccharophila S,U Copper chloride - 96 hr 96-h EC50 550 RachGreen alga,Chlorella vulgaris S,U Copper sulfate 2,000 96 hr Growth inhibition 200 YounGreen alga,Chlorella vulgaris S,U Copper chloride 33 days EC20 (growth) 42 Rosk

Green alga,Chlorella vulgaris F,U Copper sulfate - 96 hr EC50 or EC50 (cell numbers) 62 FeraGreen alga,Chlorella vulgaris S,M,D Copper sulfate - 96 hr IC50 270 FeraGreen alga,Chlorella vulgaris S,M,T Copper chloride - 96 hr EC50 (cell density) 200 Blayl

Green alga,Chlorella vulgaris S,U Copper sulfate 17.1 7 days 15% reduction in cell density 100 Bilgra

40
Table 4. Toxicity of Copper to Freshwater Plants

Species Methoda Chemical Hardness (mg/L as CaCO3) Duration Effect Resultb (Total μg/L) 

Green alga,Scenedesmus quadricauda S,U Copper sulfate 68 10 days Growth reduction 8,000 CairnGreen alga,Scenedesmus quadricauda S,U Copper sulfate 181 7 days LOEC (growth) 1,100 Bring

1979

Green alga,Selenastrum capricornutum S,U Copper chloride 14.9 14 days EC50 (cell volume) 85 ChrisGreen alga,Selenastrum capricornutum S,U Copper chloride 14.9 7 days LOEC (growth) 50 BartlGreen alga,Selenastrum capricornutum S,M,T Copper chloride 24.2 96 hr EC50 (cell count) 400 Blayl

Green alga,Selenastrum capricornutum S,U Copper sulfate 9.3 96 hr EC50 (cell count) 48.4 Blais

Green alga,Selenastrum capricornutum S,U Copper sulfate 9.3 96 hr EC50 (cell count) 44.3 Blais

Green alga,Selenastrum capricornutum S,U Copper sulfate 9.3 96 hr EC50 (cell count) 46.4 Blais

Green alga,Selenastrum capricornutum S,U Copper chloride 15 2-3 wk EC50 (biomass) 53.7 Turb

Green alga,Selenastrum capricornutum S,U Copper sulfate 14.9 5 days Growth reduction 58 NyhoGreen alga,Selenastrum capricornutum S,U Copper sulfate 9.3 96 hr EC50 (cell count) 69.9 St. L

Green alga,Selenastrum capricornutum S,U Copper sulfate 9.3 96 hr EC50 (cell count) 65.7 St. L

Green alga,Selenastrum capricornutum S,U Copper sulfate 24.2 96 hr EC50 (cell count) 54.4 Rade

Green alga,Selenastrum capricornutum R,U Copper sulfate 24.2 96 hr EC50 (cell count) 48.2 Rade

Green alga,Selenastrum capricornutum S,U Copper sulfate 16 96 hr EC50 (cell density) 38 Chen Algae,mixed culture S,U Copper sulfate - - Significant reduction in blue-green algae and nitrogen fixation 5 Elde

Diatom,Cyclotella meneghiniana S,U Copper sulfate 68 10 days Growth inhibition 8,000 CairnDiatom,Navicula incerta S,U Copper chloride - 96 hr EC50 10,429 RachDiatom,Nitzschia linearis - - - 5 day EC50 795-815 Acad

Patri

Diatom,Nitzschia palea - - - - Complete growth inhibition 5 Stee

1970

Duckweed,Lemna minor F - - 7 day EC50 119 WalbDuckweed,Lemna minor S,U Copper sulfate - 28 days Significant plant damage 130 Brow

41
Table 4. Toxicity of Copper to Freshwater Plants

Species Methoda Chemical Hardness (mg/L as CaCO3) Duration Effect Resultb (Total μg/L) 

Duckweed,Lemna minor S,U - 0 96 hr EC50 (frond number) 1,100 WanDuckweed,Lemna minor S,U Copper sulfate 78 96 hr EC50 (chlorophyll a reduction) 250 Elora

Duckweed,Lemna minor R,M,T Copper nitrate 39 96 hr Reduced chlorophyll production 24 TaraEurasian watermilfoil,Myriophyllum spicatum S,U - 89 32 days EC50 (root weight) 250 Stan

a S=Static; R=Renewal; F=Flow-through; M=Measured; U=Unmeasured; T=Total metal conc. measured; D=dissolved metal conc. measured.b Results are expressed as copper, not as the chemical.

42
43

9.0 REFERENCES

Allen, H.E. and D.J. Hansen. 1996. The importance of trace metal speciation to water quality

criteria. Water Environ. Res. 68:42-54.

Allen, H.E. et al. 1983. An algal assay method for determination of copper complexation capacities

of natural waters. Bull. Environ. Contam. Toxicol. 30:448.

Arthur, J.W. and E.N. Leonard. 1970. Effects of copper on Gammarus pseudolimnaeus, Physa

integra, and Campeloma decisum in soft water. J. Fish. Res. Board Can. 27:1277-1283.

ASTM. 2000. Standard guide for conducting acute toxicity tests on test materials with fishes,

macroinvertebrates, and amphibians. E729-96. In: Annual book of ASTM standards. Section 11,

Volume 5. American Society for Testing and Materials, Pennsylvania, PA.

Blaylock, B.G., M.L. Frank and J.F. McCarthy. 1985. Comparative toxicity of copper and acridine

to fish, Daphnia and algae. Environ. Toxicol. Chem. 4(1):63-71.

Bowen, H.J.M. 1985. In D. Hutzinger (ed.), The Handbook of Environmental Chemistry, Vol. 1,

Part D: The natural environment and biogeochemical cycles, Springer-Verlag, New York. p. 1-26.

Brown, P. L., and Markich, S. J. 2000. Evaluation of the free ion activity model of metal-organism

interaction: Extension of the conceptual model. Aquatic Toxicology 51:177-194.

Brungs, W.A., J.R. Geckler and M. Gast. 1976. Acute and chronic toxicity of copper to the

fathead minnow in a surface water of variable quality. Water Res. 10:37-43.

Bryan, S. E., Tipping, E., and Hamilton-Taylor, J. 2002. Comparison of measured and modelled

copper binding by natural organic matter in freshwaters. Comparative Biochemistry and

Physiology 133C:37-49.

Buckley, J. A., Yoshida, G. A., and Wells, N. R. 1984. A cupric ion-copper bioaccumulation

relationship in coho salmon exposed to copper-containing treated sewage. Comparative

Biochemistry and Physiology 78C:105-110.

Campbell, P.G.C. 1995. “Interactions Between Trace Metals and Aquatic Organisms: A Critique

of the Free-ion Activity Model,” in Metal Speciation and Bioavailability in Aquatic Systems, A.

Tessier and D.R. Turner, eds., John Wiley and Sons, New York, pp. 45-102.

Carlson, A.R., W.A. Brungs, G.A. Chapman and D.J. Hansen. 1984. Guidelines for deriving

numerical aquatic site-specific water quality criteria by modifying national criteria. EPA-600/3-84-

099, U.S. Environmental Protection Agency, Environmental Research Laboratory, Duluth,

Minnesota, USA.

Chakoumakos, C., R.C. Russo and R.V. Thurston. 1979. The toxicity of copper to cutthroat trout

(Salmo clarki) under different conditions of alkalinity, pH, and hardness. Environ. Sci. Technol.

13:213-219.

Chapman, G.A. 1978. Toxicities of cadmium, copper, and zinc to four juvenile stages of chinook

salmon and steelhead. Trans. Am. Fish. Soc. 107:841-847.

Davis, A. and D. Ashenberg. 1989. The aqueous geochemistry of the Berkeley Pit, Butte,

Montana, U.S.A. Appl. Geochem. Vol (4):23-36.
44

Di Toro, D.M. et al. 2001. The persistence and availability of metals in aquatic environments.

International council on metals and the environment. Ottawa, Ontario, Canada. 

Erickson, R.J., D.A. Benoit and V.R. Mattson. 1996a. A prototype toxicity factors model for sitespecific copper water quality criteria. U.S. EPA, Duluth, Minnesota.

Erickson, R.J., D.A. Benoit, V.R. Mattson, H.P. Nelson, Jr. and E.N. Leonard. 1996b. The effects

of water chemistry on the toxicity of copper to fathead minnows. Environ. Toxicol. Chem.

15(2):181-193.

Erickson, R.J., D.A. Benoit and V.R. Mattson, 1987. “A Prototype Toxicity Factors Model

ForSite-Specific Copper Water Quality Criteria,” revised September 5, 1996, United States

Environmental Protection Agency, Environmental Research Laboratory-Duluth, Duluth, MN.

Foster, P.L. 1982. Metal resistances of chlorophyta from rivers polluted by heavy metals.

Freshwater Biol. 12:41.

Gächter, R., K. Lum-Shue-Chan and Y.K Chau. 1973. Complexing capacity of the nutrient

medium and its relation to inhibition of algal photosynthesis by copper. Schweiz. Z. Hydrol.

35:252.

Geckler, J.R., W.B. Horning, T.M. Neiheisel, Q.H. Pickering, E.L. Robinson and C.E. Stephan.

1976. Validity of laboratory tests for predicting copper toxicity in streams. EPA-600/3-76-116.

National Technical Information Service, Springfield, VA.

Hem, J.D. 1989. Study and interpretation of the chemical characteristics of natural water, 3rd ed.

U.S. Geological Survey water-supply paper 2253. Government Printing Office. 

Hughes, M.M., M.A. Heber, G.E. Morrison, S.C. Schimmel and W.J. Berry. 1989. An evaluation

of a short-term chronic effluent toxicity test using sheepshead minnow (Cyprinodon variegatus)

larvae. Environ. Pollut. 60(1):1-14.

Hunt, D. T. E. 1987. Trace Metal Speciation and Toxicity to Aquatic Organisms - A Review. TR

247, Water Research Centre, Marlow, Bucks, United Kingdom. 

Inglis, A. and E.L. Davis. 1972. Effects of water hardness on the toxicity of several organic and

inorganic herbicides to fish. Technical Paper No. 67. U.S. Fish and Wildlife Service, Washington,

D.C.

Janes, N., Playle, R.C. 1995. Modeling silver binding to gills of rainbow trout (Oncorhynchus

mykiss). Env. Toxicol. Chem. 14, 1847-1858.

Kapustka, L.A., W.H. Clements, L. Ziccardi, P.R. Paquin, M. Sprenger and D. Wall, August 19,

2004. "Issue Paper on the Ecological Effects of Metals," Submitted by ERG to U.S.

Environmental Protection Agency, Risk Assessment Forum, Washington, DC.

Koivisto, S., M. Ketola and M. Walls. 1992. Comparison of five cladoceran species in short- and

long-term copper exposure. Hydrobiol. 248(2):125-136.

Kramer, J. R., Allen, H. E., Davison, W., Godtfredsen, K. L., Meyer, J. S., Perdue, E. M.,

Tipping, E., van der Meent, D., and Westall, J. C. 1997. Chemical speciation and metal toxicity in

surface freshwater. In Bergman, H. L., and Dorward-King, E. J. (eds.) Reassessment of Metals

Criteria for Aquatic Life Protection. Pensacola, FL: SETAC Press.
45

Lauren, D. J., and McDonald, D. G. 1986. Influence of water hardness, pH, and alkalinity on the

mechanisms of copper toxicity in juvenile rainbow trout, Salmo gairdneri. Canadian Journal of

Fisheries and Aquatic Sciences 43:1488-1496.

Lopez, J.M., Lee, G.F. 1977. Water, Air and Soils Pollut. Vol. (8): 373.

Lumsden, B.R. and T.M. Florence. 1983. A new algal assay procedure for the determination of the

toxicity of copper species in seawater. Environ. Toxicol. Lett. 4:271.

MacRae, R.K., D.E. Smith, N. Swoboda-Colberg, J.S. Meyer and H.L. Bergman. 1999. Copper

binding affinity of rainbow trout (Oncorhynchus mykiss) and brook trout (Salvelinus fontinalis)

gills: Implications for assessing bioavailable metal. Environ. Toxicol. Chem. 18:1180-1189.

McGeer JC, Szebedinszky C, McDonald DG, and Wood CM. 2002. The role of dissolved

organic carbon in moderating the bioavailability and toxicity of Cu to rainbow trout during chronic

waterbrone exposure. Comp. Biochem. Physiol. C: Toxicol and Pharmacol. 133C:147-161.

McKnight, D.M. and F.M.M. Morel. 1979. Release of weak and strong copper-complexing agents

by algae. Limnol. Oceanogr. 24:823.

McKnight, D.M. et al. 1983. CuSO4 treatment of nuisance algal blooms in drinking water

reservoirs. Environ. Manage. 7:311.

Meyer, J.S., C.J. Boese and S.A. Collyard. 2002. Whole-body accumulation of copper predicts

acute toxicity to an aquatic oligochaete (Lumbriculus variegatus) as pH and calcium are varied.

Comp. Biochem. Physiol. Part C 133:99-109.

Meyer, J.S., R.C. Santore, J.P. Bobbitt, L.D. DeBrey, C.J. Boese, P.R. Paquin, H.E. Allen, H.L.

Bergman and D.M. Di Toro. 1999. Binding of nickel and copper to fish gills predicts toxicity when

water hardness varies, but free-ion activity does not. Environ. Sci. Technol. 33:913-916.

Miller, T.G. and W.C. MacKay. 1980. The effects of hardness, alkalinity and pH of test water on

the toxicity of copper to rainbow trout (Salmo gairdneri). Water Res. 14:129-133.

Morel, F.M., 1983a. “Complexation: Trace Metals and Microorganisms,” in Chapter 6 of

Principles of Aquatic Chemistry, Wiley Interscience, New York, pp. 301-308.

Morel F.M.M. 1983b. Principles of Aquatic Chemistry. John Wiley and Sons, New York, NY.

Morrison, G., E. Torello, R. Comeleo, R. Walsh, A. Kuhn, R. Burgess, M. Tagliabue and W.

Greene. 1989. Intralaboratory precision of saltwater short-term chronic toxicity tests. J. Water

Pollut. Control Fed. 61(11-12):1707-1710.

Muramoto, S. 1980. Effect of complexants (EDTA, NTA and DTPA) on the exposure to high

concentrations of cadmium, copper, zinc and lead. Bull Environ Contam Toxicol. 25(6):941-946.

National Research Council. 2003. Bioavailability of Contaminants in Soils and Sediments:

Processes, Tools, and Applications. The National Academy of Sciences, Washington, DC, USA.

Nriagu, J.O. (Ed.) 1979. Copper in the Environment. Part I: Ecological Cycling; Part II: Health

Effects. Wiley and Sons, Inc. New York, NY.

Oris, J.T., R.W. Winner and M.V. Moore. 1991. A four-day survival and reproduction toxicity test

for Ceriodaphnia dubia. Environ. Toxicol. Chem. 10(2):217-224.
46

Pagenkopf, G.K. 1983. Gill surface interaction model for trace-metal toxicity to fishes: Role of

complexation, pH, and water hardness. Environ. Sci. Technol. 17:342-347.

Paquin, P.R., J.W. Gorsuch, S. Apte, G.E. Batley, K.C. Bowles, P.G.C. Campbell, C.G. Delos,

D.M. Di Toro, R.L. Dwyer, F. Galvez, R.W. Gensemer, G.G. Goss, C. Hogstrand, C.R. Janssen,

J.C. McGeer, R.B. Naddy, R.C. Playle, R.C. Santore, U. Schneider, W.A. Stubblefield, C.M.

Wood and K.B. Wu. 2002. The bitoic ligand model: A historical overview. Comp. Biochem.

Physiol. Part C 133:3-35.

Patterson, J.W., R.A. Minear, E. Gasca and C. Petropoulou. 1998. Industrial discharges of metals

to water. In: H.E. Allen, A.W. Garrison and G.W. Luther III (Eds.). Metals in Surface Waters.

Ann Arbor Press, Chelsea, MI. pp. 37-66.

Petersen, R. 1982. Influence of copper and zinc on the growth of a freshwater algae, Scenedesmus

quadricauda: The significance of speciation. Environ. Sci. Technol. 16:443.

Peterson, H. G., Healey, F. P., and Wagemann, R. 1984. Metal toxicity to algae: A highly pH

dependent phenomenon. Canadian Journal of Fisheries and Aquatic Science 41:974-979.

Playle, R.C., D.G. Dixon and K. Burnison. 1993a. Copper and cadmium binding to fish gills:

Estimates of metal-gill stability constants and modeling of metal accumulation. Can. J. Fish. Aquat.

Sci. 50(12):2678-2687.

Playle, R.C., D.G. Dixon and K. Burnison. 1993b. Copper and cadmium binding to fish gills:

Modification by dissolved organic carbon and synthetic ligands. Can. J. Fish. Aquat. Sci.

50(12):2667-2677.

Playle, R.C., R.W. Gensener and D.G. Dixon. 1992. Copper accumulation on gills of fathead

minnows: Influence of water hardness, complexation and pH of the gill micro-environment.

Environ. Toxicol. Chem. 11(3):381-391.

Reeve, W.R. et al. 1976. A controlled environmental pollution experiment (CEPEX) and its

usefulness in the study of larger marine zooplankton under toxic stress. In: Effects of pollutants on

aquatic organisms. Lockwood, P.M. (Ed.). Cambridge University Press, New York, NY. p. 145.

Robins, R.G., Berg, R.B., Dysinger, D.K., Duaime, T.E., Metesh, J.J., Diebold, F.E., Twidwell,

L.G., Mitman, G.G., Chatham , W.H., Huang, H.H., Young, C.A. 1997. Chemical, physical and

biological interactions at the Berkeley Pit, Butte, Montana. Tailings and Mine Waste 97. Bakeman,

Rotterdam.

Rueter, J.G. 1983. Alkaline phosphatase inhibition by copper: Implications to phosphorus nutrition

and use as a biochemical marker of toxicity. Limnol. Oceanogr. 28:743.

Santore, R.C., D.M. Di Toro and P.R. Paquin, H.E. Allen, and J.S. Meyer. 2001. “A Biotic Ligand

Model of the Acute Toxicity of Metals. II. Application to Acute Copper Toxicity in Freshwater

Fish and Daphnia,” Environmental Toxicology and Chemistry. 20(10):2397-2402.

Schafer, H., A. Wentzel, U. Fritsche, G. Roderer and W. Trauspurger. 1993. Long-term effects of

selected xenobiotica on freshwater green alga: Developmental of a flow-through test system. Sci.

Total Environ. Suppl.:735-740.

Smith, D. S., Bell, R. A., and Kramer, J. R. 2002. Metal speciation in natural waters with emphasis

on reduced sulfur groups as strong metal binding sites. Comparative Biochemistry and Physiology

133C:65-74.
47

Sprague, J.B. 1968. Avoidance reactions of rainbow trout to zinc sulphate solutions. Water

Res.2(5):367-372.

Steemann-Nielsen, E. and L. Kamp-Nielsen. 1970. Influence of deleterious concentrations of

copper on the growth of Chlorella pyrenoidosa. Physiol. Plant. 23:828-840.

Steemann-Nielsen, E. and S. Wium-Andersen. 1970. Copper ions as poison in sea and in

freshwater. Mar. Biol. 6:93.

Stephan, C.E. 1995. Derivation of conversation factors for the calculation of dissolved freshwater

aquatic life criteria for metals. Report. March 11, 1995. U.S. EPA, Duluth, MN.

Stephan, C.E., D.I. Mount, D.J. Hansen, J.H. Gentile, G.A. Chapman and W.A. Brungs. 1985.

Guidelines for deriving numerical national water quality criteria for the protection of aquatic

organisms and their uses. PB85-227049. National Technical Information Service, Springfield, VA.

Stephan, C.E., and J.W. Rogers. 1985. Advantages of Using Regression Analysis to Calculate

Results of Chronic Toxicity Tests. In: Aquatic Toxicology and Hazard Assessment: Eighth

Symposium. R.C. Bahner and D.J. Hansen, Eds. ASTM STP 891. American Society for Testing

and Materials, Philadelphia, PA. Pp. 328-338.

Stokes, P. and T.C. Hutchinson. 1976. Copper toxicity to phytoplankton, as affected by organic

ligands, other cations and inherent tolerance of algae to copper. In: Toxicity to biota of metal

forms in natural water. Andrew, 

Swallow, K.C. et al. 1978. Potentiometric determination of copper complexation by phytoplankton

exudates. Limnol. Oceanogr. 23:538.

Taraldsen, J.E. and T.J. Norberg-King. 1990. New method for determining effluent toxicity using

duckweed (Lemna minor). Environ. Toxicol. Chem. 9(6):761-767.

Tipping, E., 1994. “WHAM--A Chemical Equilibrium Model and Computer Code for Waters,

Sediments, and Soils Incorporating a Discrete Site/Electrostatic Model of Ion-Binding by Humic

Substances,” Computers and Geosciences, 20(6): 973-1023.

U.S. EPA. 1980. Ambient water quality criteria for copper. EPA-440/4-80-036. National

Technical Information Service, Springfield, VA.

U.S. EPA. 1983a. Methods for chemical analysis of water and wastes. EPA-600/4-79-020.

National Technical Information Service, Springfield, VA.

U.S. EPA. 1983b. Water quality standards regulation. Federal Register 48:51400. November 8.

U.S. EPA. 1983c. Water quality standards handbook. Office of Water Regulations and Standards,

Washington, D.C.

U.S. EPA. 1985. “Ambient Water Quality Criteria for Copper - 1984,” Office of Water

Regulations and Standards, Criteria and Standards Division, Washington, DC. 

U.S. EPA. 1986. Quality Criteria for Water 1986. EPA 440/5-86-001. Office of Water, Regulation

and Standards, Washington, DC. 

U.S. EPA. 1988. Short-term methods for estimating the chronic toxicity of effluents and receiving

waters to marine and estuarine organisms. EPA/600/4-87-028. National Technical Information

Service, Springfield, VA.
48

U.S. EPA. 1991. Technical support document for water quality-based toxics control. EPA-505/2-

90-001. National Technical Information Service, Springfield, VA.

U.S. EPA. 1992. Interim guidance on interpretation and implementation of aquatic life criteria for

metals. Health and Ecological Criteria Division, Office of Science and Technology, U.S.

Environmental Protection Agency, Washington, District of Columbia, USA.

U.S. EPA. 1993. Methods for measuring the acute toxicity of effluents and receiving water to

freshwater and marine organisms. Fourth Edition. EPA/600/4-90/027F. National Technical

Information Service, Springfield, VA.

U.S. EPA. 1994. Water quality standards handbook. 2nd Edition. EPA-823-B94-005b.National

Technical Information Service, Springfield, VA.

U.S. EPA. 1996 1995 Updates: Water Quality Criteria Documents for the Protection of Aquatic

Life in Ambient Water. Office of Water. EPA-820-B-96-001.

University of Wisconsin-Superior. 1995. Results of freshwater simulation tests concerning

dissolved metal. Report. March 10, 1995. U.S. EPA, Duluth, MN.

van den Berg, C.M.G., P.T.S. Wong and Y.K. Chan. 1979. Measurement of complexing materials

excreted from algae and their ability to ameliorate copper toxicity. J. Fish. Res. Board Can.

36:901-905.

Wallen, I.E., W.C. Greer and R. Lasater. 1957. Toxicity to Gambusia affinis of certain pure

chemicals in turbid waters. Sewage Ind. Wastes 29:695-711.

Winner, R.W. 1985. Bioaccumulation and toxicity of copper as affected by interactions between

humic acid and water hardness. Water Res. 19(4):449-455.

Winner, R.W. and H.A. Owen. 1991b. Toxicity of copper to Chlamydomonas reinhardtii

(Chlorophyceae) and Ceriodaphnia dubia (Crustacea) in relation to changes in water chemistry of

a freshwater pond. Aquat. Toxicol. 21(3-4):157-169.

Wood, C. M. 1992. Flux measurements as indices of H+ and metal effects on freshwater fish.

Aquatic Toxicology 22:239-264.

Wood, C. M., Adams, W. J., Ankley, G. T., DiBona, D. R., Luoma, W. N., Playle, R. C.,

Stubblefield, W. A., Bergman, H. L., Erickson, R. J., Mattice, J. S., and Schlekat, C. E. 1997.

Environmental toxicology of metals. In Bergman, H. L., and Dorward-King, E. J. (eds.)

Reassessment of Metals Criteria for Aquatic Life Protection. Pensacola, FL: SETAC Press.

Zitko, V. 1976. Toxicity to biota of metal forms in natural water. In Andrew, R. A., Hodson, P.

V., and Konasewich, D. (eds.) Toxicity to Biota of Metal Forms in Natural Water. Proceedings of

Workshop, October 1975, Duluth, MN. Windsor, Ontario: International Joint Commission.

Zitko, V., and Carson, W. G. 1976. A mechanism of the effects of water hardness on the lethality

of heavy metals to fish. Chemosphere 5:299-303.
Appendices
Appendix A. Ranges in Calibration and Application Data Sets
0

10203040

Temperature (

oC)

46 0

Clark Fork River

33 0

Penn. Copper Group

3 0

*Schubauer-Ber., 199

1 0

*Spehar, 1986

1 0

*Oris, 1991

1 0

*Diamond, 1997b

24 0

*Lind, 1978

13 0

*Lazorchak, 1993

10 0

*Meador, 1991

4 0

*Chapman, 1980

0 1

*Borgmann, 1983

0 4

*Borgmann, 1984

6 0

*Baird, 1991

0 17

Dunbar, 1996

20 0

*Winner, 1985

3 0

*Schubauer-Ber., 199

0 7

*Collyard, 1994

4 0

*Welsh, 1996

30 0

Clark Fork River

1050

Erickson, 1996

0 13

Diamond, 1997a

0 13

Dunbar, 1996

8 0

*Hagler-Bailly, 1996

4 0

*Chapman, 1975, 1978

0

10203040

Temperature (

oC)

N

___ Measured/Nominal/Calculated Data___ Assumed/Untraceable Data

C. dubia --------------> D.

pulicaria D. magna -----------> D. pulex H. azteca F. minnow -> R. trou

Median, Range and Quartiles of Temperature in BLM Calibration and Appl

(All species, Median and Quartiles calculated directly from data i.e., no distribu

A-1
0

102030405060708090

100

HA (%)

0 46

Clark Fork River

0 33

Penn. Copper Group

0 3

*Schubauer-Ber., 199

0 1

*Spehar, 1986

0 1

*Oris, 1991

0 1

*Diamond, 1997b

0 24

*Lind, 1978

0 13

*Lazorchak, 1993

0 10

*Meador, 1991

0 4

*Chapman, 1980

0 1

*Borgmann, 1983

0 4

*Borgmann, 1984

0 6

*Baird, 1991

0 17

Dunbar, 1996

0 20

*Winner, 1985

0 3

*Schubauer-Ber., 199

0 7

*Collyard, 1994

0 4

*Welsh, 1996

0 30

Clark Fork River

0105

Erickson, 1996

0 13

Diamond, 1997a

0 13

Dunbar, 1996

0 8

*Hagler-Bailly, 1996

0 4

*Chapman, 1975, 1978

0

102030405060708090

100

HA (%)

N

___ Measured/Nominal/Calculated Data___ Assumed/Untraceable Data

C. dubia --------------> D.

pulicaria D. magna -----------> D. pulex H. azteca F. minnow -> R. trou

Median, Range and Quartiles of HA in BLM Calibration and Applicatio

(All species, Median and Quartiles calculated directly from data i.e., no distribu

A-2
4

56789

10

pH

46 0

Clark Fork River

33 0

Penn. Copper Group

3 0

*Schubauer-Ber., 199

1 0

*Spehar, 1986

1 0

*Oris, 1991

1 0

*Diamond, 1997b

17 7

*Lind, 1978

13 0

*Lazorchak, 1993

10 0

*Meador, 1991

4 0

*Chapman, 1980

0 1

*Borgmann, 1983

0 4

*Borgmann, 1984

6 0

*Baird, 1991

0 17

Dunbar, 1996

18 2

*Winner, 1985

3 0

*Schubauer-Ber., 199

0 7

*Collyard, 1994

4 0

*Welsh, 1996

30 0

Clark Fork River

1050

Erickson, 1996

13 0

Diamond, 1997a

0 13

Dunbar, 1996

8 0

*Hagler-Bailly, 1996

4 0

*Chapman, 1975, 1978

456789

10

pH

N

___ Measured/Nominal/Calculated Data___ Assumed/Untraceable Data

C. dubia --------------> D.

pulicaria D. magna -----------> D. pulex H. azteca F. minnow -> R. trou

Median, Range and Quartiles of pH in BLM Calibration and Applicatio

(All species, Median and Quartiles calculated directly from data i.e., no distribu

A-3
10 -2

10 -110 010 110 2

DOC (mg/L)

2026

Clark Fork River

2013

Penn. Copper Group

0 3

*Schubauer-Ber., 199

0 1

*Spehar, 1986

0 1

*Oris, 1991

0 1

*Diamond, 1997b

17 7

*Lind, 1978

0 13

*Lazorchak, 1993

10 0

*Meador, 1991

4 0

*Chapman, 1980

0 1

*Borgmann, 1983

0 4

*Borgmann, 1984

6 0

*Baird, 1991

0 17

Dunbar, 1996

20 0

*Winner, 1985

0 3

*Schubauer-Ber., 199

0 7

*Collyard, 1994

4 0

*Welsh, 1996

1119

Clark Fork River

0105

Erickson, 1996

0 13

Diamond, 1997a

0 13

Dunbar, 1996

8 0

*Hagler-Bailly, 1996

4 0

*Chapman, 1975, 1978

10 -210 -110 010 110 2

DOC (mg/L)

N

___ Measured/Nominal/Calculated Data___ Assumed/Untraceable Data

C. dubia --------------> D.

pulicaria D. magna -----------> D. pulex H. azteca F. minnow -> R. trou

Median, Range and Quartiles of DOC in BLM Calibration and Applicat

(All species, Median and Quartiles calculated directly from data i.e., no distribu

A-4
10 -1

10 010 110 210 3

Ca (mg/L as CaCO

3

)

46 0

Clark Fork River

33 0

Penn. Copper Group

3 0

*Schubauer-Ber., 199

1 0

*Spehar, 1986

1 0

*Oris, 1991

1 0

*Diamond, 1997b

17 7

*Lind, 1978

13 0

*Lazorchak, 1993

10 0

*Meador, 1991

4 0

*Chapman, 1980

0 1

*Borgmann, 1983

0 4

*Borgmann, 1984

6 0

*Baird, 1991

0 17

Dunbar, 1996

20 0

*Winner, 1985

3 0

*Schubauer-Ber., 199

0 7

*Collyard, 1994

4 0

*Welsh, 1996

30 0

Clark Fork River

1050

Erickson, 1996

13 0

Diamond, 1997a

0 13

Dunbar, 1996

8 0

*Hagler-Bailly, 1996

4 0

*Chapman, 1975, 1978

10 -110 010 110 210 3

Ca (mg/L as CaCO

3

)

N

___ Measured/Nominal/Calculated Data___ Assumed/Untraceable Data

C. dubia --------------> D.

pulicaria D. magna -----------> D. pulex H. azteca F. minnow -> R. trou

Median, Range and Quartiles of Ca in BLM Calibration and Applicatio

(All species, Median and Quartiles calculated directly from data i.e., no distrib
10 -2

10 -110 010 110 210 3

Mg (mg/L as CaCO

3

)

46 0

Clark Fork River

33 0

Penn. Copper Group

3 0

*Schubauer-Ber., 199

1 0

*Spehar, 1986

1 0

*Oris, 1991

1 0

*Diamond, 1997b

17 7

*Lind, 1978

13 0

*Lazorchak, 1993

10 0

*Meador, 1991

4 0

*Chapman, 1980

0 1

*Borgmann, 1983

0 4

*Borgmann, 1984

6 0

*Baird, 1991

0 17

Dunbar, 1996

20 0

*Winner, 1985

3 0

*Schubauer-Ber., 199

0 7

*Collyard, 1994

4 0

*Welsh, 1996

30 0

Clark Fork River

1050

Erickson, 1996

13 0

Diamond, 1997a

0 13

Dunbar, 1996

8 0

*Hagler-Bailly, 1996

4 0

*Chapman, 1975, 1978

0

102030405060708090

100

HA (%)

N

___ Measured/Nominal/Calculated Data___ Assumed/Untraceable Data

C. dubia --------------> D.

pulicaria D. magna -----------> D. pulex H. azteca F. minnow -> R. tro

Median, Range and Quartiles of HA in BLM Calibration and Applicat

(All species, Median and Quartiles calculated directly from data i.e., no distrib

A-6
10 -1

10 010 110 210 3

Na (mg/L as Na)

46 0

Clark Fork River

1320

Penn. Copper Group

3 0

*Schubauer-Ber., 199

1 0

*Spehar, 1986

1 0

*Oris, 1991

1 0

*Diamond, 1997b

17 7

*Lind, 1978

13 0

*Lazorchak, 1993

10 0

*Meador, 1991

4 0

*Chapman, 1980

0 1

*Borgmann, 1983

0 4

*Borgmann, 1984

6 0

*Baird, 1991

0 17

Dunbar, 1996

20 0

*Winner, 1985

3 0

*Schubauer-Ber., 199

0 7

*Collyard, 1994

4 0

*Welsh, 1996

30 0

Clark Fork River

1050

Erickson, 1996

0 13

Diamond, 1997a

0 13

Dunbar, 1996

8 0

*Hagler-Bailly, 1996

4 0

*Chapman, 1975, 1978

10 -110 010 110 210 3

Na (mg/L as Na)

N

___ Measured/Nominal/Calculated Data___ Assumed/Untraceable Data

C. dubia --------------> D.

pulicaria D. magna -----------> D. pulex H. azteca F. minnow -> R. trou

Median, Range and Quartiles of Na in BLM Calibration and Applicatio

(All species, Median and Quartiles calculated directly from data i.e., no distribu

A-7
10 -2

10 -110 010 110 210 3

K (mg/L as K)

46 0

Clark Fork River

1320

Penn. Copper Group

3 0

*Schubauer-Ber., 199

1 0

*Spehar, 1986

1 0

*Oris, 1991

1 0

*Diamond, 1997b

17 7

*Lind, 1978

13 0

*Lazorchak, 1993

10 0

*Meador, 1991

4 0

*Chapman, 1980

0 1

*Borgmann, 1983

0 4

*Borgmann, 1984

6 0

*Baird, 1991

0 17

Dunbar, 1996

20 0

*Winner, 1985

3 0

*Schubauer-Ber., 199

0 7

*Collyard, 1994

4 0

*Welsh, 1996

30 0

Clark Fork River

1050

Erickson, 1996

0 13

Diamond, 1997a

0 13

Dunbar, 1996

8 0

*Hagler-Bailly, 1996

4 0

*Chapman, 1975, 1978

10 -210 -110 010 110 210 3

K (mg/L as K)

N

___ Measured/Nominal/Calculated Data___ Assumed/Untraceable Data

C. dubia --------------> D.

pulicaria D. magna -----------> D. pulex H. azteca F. minnow -> R. trou

Median, Range and Quartiles of K in BLM Calibration and Applicatio

(All species, Median and Quartiles calculated directly from data i.e., no distribu

A-8
10 -2

10 -110 010 110 210 3

SO

4 (mg/L as SO

4

)

46 0

Clark Fork River

1320

Penn. Copper Group

3 0

*Schubauer-Ber., 199

1 0

*Spehar, 1986

1 0

*Oris, 1991

1 0

*Diamond, 1997b

17 7

*Lind, 1978

13 0

*Lazorchak, 1993

10 0

*Meador, 1991

4 0

*Chapman, 1980

0 1

*Borgmann, 1983

0 4

*Borgmann, 1984

6 0

*Baird, 1991

0 17

Dunbar, 1996

8 12

*Winner, 1985

3 0

*Schubauer-Ber., 199

0 7

*Collyard, 1994

4 0

*Welsh, 1996

30 0

Clark Fork River

1050

Erickson, 1996

0 13

Diamond, 1997a

0 13

Dunbar, 1996

8 0

*Hagler-Bailly, 1996

4 0

*Chapman, 1975, 1978

10 -210 -110 010 110 210 3

SO

4 (mg/L as SO

4

)

N

___ Measured/Nominal/Calculated Data___ Assumed/Untraceable Data

C. dubia --------------> D.

pulicaria D. magna -----------> D. pulex H. azteca F. minnow -> R. trou

Median, Range and Quartiles of SO4 in BLM Calibration and Applicati

(All species, Median and Quartiles calculated directly from data i.e., no distribu

A-9
10 -1

10 010 110 210 3

Cl (mg/L as Cl)

3115

Clark Fork River

1320

Penn. Copper Group

3 0

*Schubauer-Ber., 199

1 0

*Spehar, 1986

1 0

*Oris, 1991

1 0

*Diamond, 1997b

17 7

*Lind, 1978

13 0

*Lazorchak, 1993

10 0

*Meador, 1991

4 0

*Chapman, 1980

0 1

*Borgmann, 1983

0 4

*Borgmann, 1984

6 0

*Baird, 1991

0 17

Dunbar, 1996

20 0

*Winner, 1985

3 0

*Schubauer-Ber., 199

0 7

*Collyard, 1994

4 0

*Welsh, 1996

1515

Clark Fork River

1050

Erickson, 1996

0 13

Diamond, 1997a

0 13

Dunbar, 1996

8 0

*Hagler-Bailly, 1996

4 0

*Chapman, 1975, 1978

10 -110 010 110 210 3

Cl (mg/L as Cl)

N

___ Measured/Nominal/Calculated Data___ Assumed/Untraceable Data

C. dubia --------------> D.

pulicaria D. magna -----------> D. pulex H. azteca F. minnow -> R. trou

Median, Range and Quartiles of Cl in BLM Calibration and Applicatio

(All species, Median and Quartiles calculated directly from data i.e., no distribu

A-10
10 0

10 110 210 3

Alkalinity (mg/L as CaCO

3

)

46 0

Clark Fork River

33 0

Penn. Copper Group

3 0

*Schubauer-Ber., 199

1 0

*Spehar, 1986

1 0

*Oris, 1991

1 0

*Diamond, 1997b

17 7

*Lind, 1978

13 0

*Lazorchak, 1993

10 0

*Meador, 1991

4 0

*Chapman, 1980

0 1

*Borgmann, 1983

0 4

*Borgmann, 1984

6 0

*Baird, 1991

0 17

Dunbar, 1996

20 0

*Winner, 1985

3 0

*Schubauer-Ber., 199

0 7

*Collyard, 1994

4 0

*Welsh, 1996

30 0

Clark Fork River

1050

Erickson, 1996

13 0

Diamond, 1997a

0 13

Dunbar, 1996

8 0

*Hagler-Bailly, 1996

4 0

*Chapman, 1975, 1978

10 010 110 210 3

Alkalinity (mg/L as CaCO

3

)

N

___ Measured/Nominal/Calculated Data___ Assumed/Untraceable Data

C. dubia --------------> D.

pulicaria D. magna -----------> D. pulex H. azteca F. minnow -> R. trou

Median, Range and Quartiles of Alkalinity in BLM Calibration and Applic

(All species, Median and Quartiles calculated directly from data i.e., no distribu

A-11
Appendix B. Other Data on Effects of Copper on 

Freshwater Organisms
Species Methoda Chemical

Hardness 

(mg/L as 

CaCO3)

Duration Effect

Total

Concentration

(μg/L)b

Dissolved

Concentration

(μg/L)

Bacteria,Escherichia coli

S,U Copper 

sulfate

- 48 hr Threshold of inhibited glucose use; 

measured by pH change in media

80 - 

Bacteria,Pseudomonas putida

S,U Copper 

sulfate

81.1 16 hr EC3

(cell numbers)

30 - 

Protozoan,Entosiphon sulcatum

S,U Copper 

sulfate

81.9 72 hr EC5

(cell numbers)

110 - 

Protozoan,Microrega heterostoma

S,U Copper 

sulfate

214 28 hr Threshold of decreased feeding rate 50 - 

Protozoan,Chilomonas paramecium

S,U Copper 

sulfate

- 48 hr Growth threshold 3,200 - 

Protozoan,Uronema parduezi

S,U Copper 

sulfate

- 20 hr Growth threshold 140 - 

Protozoa,mixed species

- - - 7 days Reduced rate of colonization 167 - 

Protozoa,mixed species

S,M,T Copper 

sulfate

- 15 days Reduced rate of colonization 100 - 

Green alga,Cladophora glomerata

Dosed 

stream

Copper 

sulfate

226-310 10 mo Decreased abundance from 21% down

to 0%

120 - 

Green alga,Chlamydomonas reinhardtii

- Copper 

sulfate

76 72 hr Deflagellation 6.7 - 

Green alga,Chlamydomonas reinhardtii

- Copper 

sulfate

76 72 hr Deflagellation 6.7 - 

Green alga,Chlamydomonas reinhardtii

- Copper 

sulfate

76 72 hr Deflagellation 16.3 - 

Green alga,Chlamydomonas reinhardti

- Copper 

sulfate

76 72 hr Deflagellation 25.4 - 

Green alga,Chlorella sp.

S,U Copper 

nitrate

- 28 hr Inhibited photosynthesis 6.3 - 

Green alga,Chlorella pyrenoidosa

S,U - 29.4 72 hr IC50

(cell division rate)

16 - 

Green alga,Chlorella pyrenoidosa

S,U - 14.9 72 hr IC50

(cell division rate)

24 - 

Green alga,Chlorella pyrenoidosa

S,U Copper 

sulfate

82 4 hr Disturbed 

photosystem II

25 - 

Green alga,Eudorina californica

S,U Copper 

sulfate

19.1 - Decrease in cell density 5,000 - 

Green alga (flagellate cells),Haematococcus sp.

S,U Copper 

sulfate

2 24 hr Inhibited growth during 96 hr recovery 

period

50 - 

Green alga,Scenedesmus quadricauda

S,U Copper 

sulfate

214 96 hr Threshold of effect on cell numbers 150 - 

Green alga,Scenedesmus quadricauda

S,U Copper 

sulfate

60 72 hr EC3

(cell numbers)

1,100 - 

Green alga,Scenedesmus quadricauda

S,U Copper 

sulfate

34.8 24 hr EC50 

(photosynthesis)

100 - 

B-1

Appendix B. Other Data on Effects of Copper on Freshwater Organisms
Species Methoda Chemical

Hardness 

(mg/L as 

CaCO3)

Duration Effect

Total

Concentration

(μg/L)b

Dissolved

Concentration

(μg/L)

Green alga,Scenedesmus quadricauda

S,U Copper 

sulfate

34.8 24 hr NOEC

(growth)

50 - 

Green alga,Scenedesmus quadricauda

S,U Copper 

sulfate

34.8 24 hr NOEC

(growth)

50 - 

Green alga,Scenedesmus quadricauda

S,U Copper 

sulfate

34.8 24 hr NOEC

(growth)

>200 - 

Green alga,Selenastrum capricornutum

S,U Copper 

chloride

14.9 7 days Growth reduction 50 - 

Green alga,Selenastrum capricornutum

S,U Copper 

sulfate

29.3 72 hr EC50

(cell count)

19 - 

Green alga,Selenastrum capricornutum

S,U Copper 

sulfate

24.2 72 hr EC50

(cell count)

41 - 

Green alga,Selenastrum capricornutum

S,U Copper 

sulfate

24.2 72 hr EC50

(cell count)

28 - 

Green alga,Selenastrum capricornutum

S,U Copper 

sulfate

14.9 72 hr EC50

(cell count)

60 - 

Green alga,Selenastrum capricornutum

S,U Copper 

sulfate

24.2 72 hr EC50

(cell count)

28.5 - 

Green alga,Selenastrum capricornutum

F,U Copper 

sulfate

15 24 hr EC50 

(cell density) 

21 - 

Diatom,Cocconeis placentula

Dosed 

stream

Copper 

sulfate

226-310 10 mo Decreased abundance from 21% down

to <1%

120 - 

Phytoplankton,mixed species

S,U - - 124 hr Averaged 39% reduction in primary 

production

10 - 

Macrophyte,Elodea canadensis

S,U Copper 

sulfate

- 24 hr EC50

(photosynthesis)

150 - 

Microcosm F,M,T,D Copper 

sulfate

200 32 wk LOEC

(primary production)

9.3 - 

Microcosm F,M,T,D Copper 

sulfate

200 32 wk NOEC

(primary production)

4 - 

Microcosm F,M,T Copper 

sulfate

76.7 96 hr Significant drop in no. of taxa and no. 

of individuals

15 - 

Microcosm F,M,T Copper 

sulfate

58.5 10 days Significant drop in no. of individuals 2.5 - 

Microcosm F,M,T Copper 

sulfate

151 10 days 58% drop in no. of individuals 13.5 - 

Microcosm F,M,T Copper 

sulfate

68 10 days Significant drop in species richness 

and no. of individuals

11.3 - 

Microcosm F,M,T Copper 

sulfate

80 10 days Significant drop in species richness 

and no. of individuals

10.7 - 

Microcosm S,M,T Copper 

sulfate

102 5 wk 14-28% drop in phytoplankton species 

richness

20 - 

Microcosm F,M,T - 160 28 days LOEC

(species richness)

19.9 - 

B-2

Appendix B. Other Data on Effects of Copper on Freshwater Organisms
Species Methoda Chemical

Hardness 

(mg/L as 

CaCO3)

Duration Effect

Total

Concentration

(μg/L)b

Dissolved

Concentration

(μg/L)

Dosed stream F,M,D Copper 

sulfate

56 1 yr Shifts in periphyton species 

abundance

5.208 - 

Dosed stream F,M,D Copper 

sulfate

56 1 yr Reduced algal production 5.208 - 

Sponge,Ephydatia fluviatilis

S,U Copper 

sulfate

200 10 days Reduced growth by 33% 6 - 

Sponge,Ephydatia fluviatilis

S,U Copper 

sulfate

200 10 days Reduced growth by 100% 19 - 

Rotifer,Philodina acuticornis

S,U Copper 

sulfate

45 48 hr LC50

(50 C)

1,300 - 

Rotifer,Philodina acuticornis

S,U Copper 

sulfate

45 48 hr LC50

(100 C)

1,200 - 

Rotifer,Philodina acuticornis

S,U Copper 

sulfate

45 48 hr LC50

(150 C)

1,130 - 

Rotifer,Philodina acuticornis

S,U Copper 

sulfate

45 48 hr LC50

(200 C)

1,000 - 

Rotifer,Philodina acuticornis

S,U Copper 

sulfate

45 48 hr LC50

(250 C)

950 - 

Rotifer,Brachionus calyciflorus

S, U Copper 

sulfate

39.8 24 hr EC50

(mobility)

200 - 

Rotifer (2 hr),Brachionus calyciflorus

S,U Copper 

sulfate

- 2 hr LOEC

(swimming activity)

12.5 - 

Rotifer,Brachionus calyciflorus

S,U Copper 

sulfate

90 24 hr EC50

(mobility)

76 - 

Rotifer (2 hr),Brachionus calyciflorus

S,U Copper 

sulfate

90 5 hr EC50 

(filtration rate)

34 - 

Rotifer (2 hr),Brachionus calyciflorus

S,U Copper 

sulfate

90 6 days LOEC 

(reproduction decreased 26%)

5 - 

Rotifer (2 hr),Brachionus calyciflorus

S,U Copper 

sulfate

90 5 hr LOEC 

(reduced swimming speed)

12 - 

Rotifer (2 hr),Brachionus calyciflorus

S,U Copper 

sulfate

85 3 days LOEC

(reproduction decreased 27%) 

5 - 

Rotifer (2 hr),Brachionus calyciflorus

S,U Copper 

sulfate

85 3 days LOEC

(reproduction decreased 29%) 

5 - 

Rotifer (2 hr),Brachionus calyciflorus

S,U Copper 

sulfate

85 8 days LOEC 

(reproduction decreased 47%)

5 - 

Rotifer (2 hr),Brachionus calyciflorus

S,U Copper 

chloride

170 35 min LOEC 

(food ingestion rate)

100 - 

Rotifer (2 hr),Brachionus calyciflorus

S,U Copper 

sulfate

63.2 24 hr EC50

(mobility)

9.4 - 

Rotifer (2 hr),Brachionus calyciflorus

S,U - 90 2 days LOEC

(reproduction decreased 100%)

30 - 

Rotifer (<2 hr),Brachionus calyciflorus

S, U - 85 24 hr EC50

(mobility)

26 - 

B-3

Appendix B. Other Data on Effects of Copper on Freshwater Organisms
Species Methoda Chemical

Hardness 

(mg/L as 

CaCO3)

Duration Effect

Total

Concentration

(μg/L)b

Dissolved

Concentration

(μg/L)

Rotifer (<2 hr),Brachionus calyciflorus

S, U - 85 24 hr EC50 

(mobility; 100 C)

18 - 

Rotifer (<2 hr),Brachionus calyciflorus

S, U - 85 24 hr EC50

(mobility; 150 C)

31 - 

Rotifer (<2 hr),Brachionus calyciflorus

S, U - 85 24 hr EC50

(mobility; 200 C)

31 - 

Rotifer (<2 hr),Brachionus calyciflorus

S, U - 85 24 hr EC50

(mobility; 250 C)

26 - 

Rotifer (<2 hr),Brachionus calyciflorus

S, U - 85 24 hr EC50

(mobility; 300 C)

25 - 

Rotifer (<3 hr),Brachionus rubens

S, U Copper 

sulfate

90 24 hr LC50 19 - 

Rotifer,Keratella cochlearis

S,U Copper 

chloride

- 24 hr LC50 101 - 

Worm,Aeolosoma headleyi

S,U Copper 

sulfate

45 48 hr LC50

(50 C)

2,600 - 

Worm,Aeolosoma headleyi

S,U Copper 

sulfate

45 48 hr LC50

(100 C)

2,300 - 

Worm,Aeolosoma headleyi

S,U Copper 

sulfate

45 48 hr LC50

(150 C)

2,000 - 

Worm,Aeolosoma headleyi

S,U Copper 

sulfate

45 48 hr LC50

(200 C)

1,650 - 

Worm,Aeolosoma headleyi

S,U Copper 

sulfate

45 48 hr LC50

(50 C)

1,000 - 

Worm (adult),Lumbriculus variegatus

S,U Copper 

sulfate

30 LC50 150 

Worm (7 mg),Lumbriculus variegatis

F,M,T Copper 

sulfate

45 10 days LC50 35 - 

Tubificid worm,Limnodrilus hoffmeisteri

S,U Copper 

sulfate

100 LC50 102 

Tubificid worm,Tubifex tubifex

R, U Copper 

sulfate

245 LC50 158 

Snail (11-27 mm),Campeloma decisum

F,M,T Copper 

sulfate

45 6 wk LOEC

(mortality)

14.8 - 

Snail,Gyraulus circumstriatus

S,U Copper 

sulfate

100 LC50 108 

Snail,Goniobasis livescens

S,U Copper 

sulfate

154 48 hr LC50 860 - 

Snail,Goniobasis livescens

S,M,D Copper 

sulfate

154 96 hr LC50 - 390 

Snail,Nitrocris sp.

S,U Copper 

sulfate

45 48 hr LC50

(50 C)

3,000 - 

Snail,Nitrocris sp.

S,U Copper 

sulfate

45 48 hr LC50

(100 C)

2,400 - 

B-4

Appendix B. Other Data on Effects of Copper on Freshwater Organisms
Species Methoda Chemical

Hardness 

(mg/L as 

CaCO3)

Duration Effect

Total

Concentration

(μg/L)b

Dissolved

Concentration

(μg/L)

Snail,Nitrocris sp.

S,U Copper 

sulfate

45 48 hr LC50

(150 C)

1,000 - 

Snail,Nitrocris sp.

S,U Copper 

sulfate

45 48 hr LC50

(200 C)

300 - 

Snail,Nitrocris sp.

S,U Copper 

sulfate

45 48 hr LC50 

(250 C)

210 - 

Snail,Lymnaea emarginata

S,U Copper 

sulfate

154 48 hr LC50 300 - 

Snail (adult),Juga plicifera

F,M,T Copper 

chloride

23 30 days LC50 6 - 

Snail (adult),Lithoglyphus virens

F,M,T Copper 

chloride

23 30 days LC50 4 - 

Snail,Physa heterostropha

S,U Copper 

sulfate

100 LC50 69 

Freshwater mussel (released glochidia),Actinonaias pectorosa

R,M Copper 

sulfate

140 24 hr 132 

Freshwater mussel (released glochidia),Actinonaias pectorosa

R,M Copper 

sulfate

150 24 hr 93 

Freshwater mussel (released glochidia),Actinonaias pectorosa

R,M Copper 

sulfate

170 24 hr 67 

Freshwater mussel (released glochidia),Actinonaias pectorosa

R,M Copper 

sulfate

140 24 hr 42 

Freshwater mussel (released glochidia),Actinonaias pectorosa

R,M Copper 

sulfate

170 48 hr 51 

Freshwater mussel (1-2 d),Anodonta grandis

S,M,T Copper 

sulfate

70 24 hr LC50 44 - 

Freshwater mussel (1-2 d), Anodonta imbecilis

S,M,T Copper 

sulfate

39 48 hr LC50 171 - 

Freshwater mussel (1-2 d), Anodonta imbecilis

S,M,T Copper 

sulfate

90 48 hr LC50 388 - 

Freshwater mussel (released glochidia), Lampsilis fasciola

R,M,T Copper 

sulfate

170 24 hr 48 

Freshwater mussel (released glochidia), Lampsilis fasciola

R,M,T Copper 

sulfate

160 24 hr 26 

Freshwater mussel (released glochidia), Lampsilis fasciola

R,M,T Copper 

sulfate

75 24 hr 46 

B-5

Appendix B. Other Data on Effects of Copper on Freshwater Organisms
Species Methoda Chemical

Hardness 

(mg/L as 

CaCO3)

Duration Effect

Total

Concentration

(μg/L)b

Dissolved

Concentration

(μg/L)

Freshwater mussel (released glochidia), Lampsilis fasciola

R,M,T Copper 

sulfate

170 48 hr 40 

Freshwater mussel (released glochidia),Medionidus conradicus

R,M,T Copper 

sulfate

185 24 hr 69 

Freshwater mussel (released glochidia),Medionidus conradicus

R,M,T Copper 

sulfate

185 24 hr 40 

Freshwater mussel (released glochidia),Medionidus conradicus

R,M,T Copper 

sulfate

185 24 hr 37 

Freshwater mussel (released glochidia),Medionidus conradicus

R,M,T Copper 

sulfate

170 24 hr 46 

Freshwater mussel (released glochidia),Medionidus conradicus

R,M,T Copper 

sulfate

160 24 hr 41 

Freshwater mussel (released glochidia),Medionidus conradicus

R,M,T Copper 

sulfate

150 24 hr 81 

Freshwater mussel (released glochidia),Medionidus conradicus

R,M,T Copper 

sulfate

170 48 hr 16 

Freshwater mussel (released glochidia),Pygranodon grandis

R,M,T Copper 

sulfate

170 24 hr >160 

Freshwater mussel (released glochidia),Pygranodon grandis

R,M,T Copper 

sulfate

170 24 hr 347 

Freshwater mussel (released glochidia),Pygranodon grandis

R,M,T Copper 

sulfate

50 24 hr 46 

Freshwater mussel (1-2 d),Villosa iris

S,M,T Copper 

sulfate

190 24 hr LC50 83 - 

Freshwater mussel (released glochidia),Villosa iris

R,M,T Copper 

sulfate

190 24 hr 80 

Freshwater mussel (released glochidia),Villosa iris

R,M,T Copper 

sulfate

190 24 hr 73 

Freshwater mussel (released glochidia),Villosa iris

R,M,T Copper 

sulfate

185 24 hr 65 

B-6

Appendix B. Other Data on Effects of Copper on Freshwater Organisms
Species Methoda Chemical

Hardness 

(mg/L as 

CaCO3)

Duration Effect

Total

Concentration

(μg/L)b

Dissolved

Concentration

(μg/L)

Freshwater mussel (released glochidia),Villosa iris

R,M,T Copper 

sulfate

185 24 hr 46 

Freshwater mussel (released glochidia),Villosa iris

R,M,T Copper 

sulfate

170 24 hr 75 

Freshwater mussel (released glochidia),Villosa iris

R,M,T Copper 

sulfate

160 24 hr 46 

Freshwater mussel (released glochidia),Villosa iris

R,M,T Copper 

sulfate

160 24 hr 36 

Freshwater mussel (released glochidia),Villosa iris

R,M,T Copper 

sulfate

155 24 hr 39 

Freshwater mussel (released glochidia),Villosa iris

R,M,T Copper 

sulfate

155 24 hr 37 

Freshwater mussel (released glochidia),Villosa iris

R,M,T Copper 

sulfate

150 24 hr 46 

Freshwater mussel (released glochidia),Villosa iris

R,M,T Copper 

sulfate

150 24 hr 46 

Freshwater mussel (released glochidia),Villosa iris

R,M,T Copper 

sulfate

55 24 hr 55 

Freshwater mussel (released glochidia),Villosa iris

R,M,T Copper 

sulfate

55 24 hr 38 

Freshwater mussel (released glochidia),Villosa iris

R,M,T Copper 

sulfate

50 24 hr 71 

Freshwater mussel (released glochidia),Villosa iris

R,M,T Copper 

sulfate

160 24 hr 46 

Freshwater mussel (released glochidia),Villosa iris

R,M,T Copper 

sulfate

170 48 hr 66 

Freshwater mussel (released glochidia),Villosa iris

R,M,T Copper 

sulfate

150 48 hr 46 

Zebra mussel (1.6-2.0 cm),Dreissena polymorpha

R,M,T Copper 

chloride

268 9 wk EC50 

+F106(filtration rate)

43 - 

B-7

Appendix B. Other Data on Effects of Copper on Freshwater Organisms
Species Methoda Chemical

Hardness 

(mg/L as 

CaCO3)

Duration Effect

Total

Concentration

(μg/L)b

Dissolved

Concentration

(μg/L)

Zebra mussel (1.6-2.0 cm),Dreissena polymorpha

R,M,T Copper 

chloride

268 10 wk NOEC 

(filtration rate)

13 - 

Asiatic clam (1.0-2.1 cm),Coprbicula fluminea

S,M,T Copper 

sulfate

64 96 hr (24hr 

LC50 also 

reported)

LC50 40 - 

Asiatic clam (1.0-2.1 cm),Coprbicula fluminea

F,M,T Copper 

sulfate

64 96 hr (24 hr 

LC50 also 

reported)

LC50 490 - 

Asiatic clam (juvenile),Corbicula fluminea 

F,M,D Copper 

sulfate

78 30 days 43.3% mortality 14.48 - 

Asiatic clam (juvenile),Corbicula fluminea 

F,M,D Copper 

sulfate

78 30 days Stopped shell growth 8.75 - 

Asiatic clam (adult),Corbicula fluminea 

F,M,D Copper 

sulfate

78 30 days 13.3% mortality 14.48 - 

Asiatic clam (adult),Corbicula fluminea 

F,M,D Copper 

sulfate

71 30 days 25% mortality 16.88 - 

Asiatic clam (adult),Corbicula fluminea 

F,M,D Copper 

sulfate

78 30 days Inhibited shell growth 8.75 - 

Asiatic clam (adult),Corbicula fluminea 

F,M,D Copper 

sulfate

- 15-16 days LC50 - - 

Asiatic clam (adult),Corbicula fluminea 

F,M,D Copper 

sulfate

- 19 days LC100 - - 

Asiatic clam (veliger larva),Corbicula manilensis

S,M,T Copper 

chloride

- 24 hr 34% mortality 10 - 

Asiatic clam (juvenile),Corbicula manilensis

S,M,T Copper 

chloride

17 24 hr LC50 100 - 

Asiatic clam (veliger),Corbicula manilensis

S,M,T Copper 

chloride

17 24 hr LC50 28 - 

Asiatic clam (trochophore),Corbicula manilensis

S,M,T Copper 

chloride

17 8 hr LC100 7.7 - 

Asiatic clam (adult),Corbicula manilensis

F,M,T Copper 

chloride

17 7 days LC50 3,638 - 

Asiatic clam (adult),Corbicula manilensis

F,M,T Copper 

chloride

17 42 days LC50 12 - 

Asiatic clam (4.3 g adult),Corbicula manilensis

F,M,T Copper 

chloride

17 30 days LC50 11 - 

Cladoceran,Bosmina longirostrus

S, U Copper 

sulfate

33.8 EC50 1.6 

Cladoceran (<24 hr),Daphnia ambigua

S,U Copper 

sulfate

145 72 hr LC50 86.5 - 

Cladoceran (<24 hr),Daphnia ambigua

S,U Copper 

sulfate

145 Life span 

(ca. 5 wk)

Chronic limits (inst. rate of population 

growth)

50 - 

Cladoceran,Ceriodaphnia dubia

S,U Copper 

sulfate

188 EC50 36.6 

B-8

Appendix B. Other Data on Effects of Copper on Freshwater Organisms
Species Methoda Chemical

Hardness 

(mg/L as 

CaCO3)

Duration Effect

Total

Concentration

(μg/L)b

Dissolved

Concentration

(μg/L)

Cladoceran,Ceriodaphnia dubia

S,U Copper 

sulfate

204 EC50 19.1 

Cladoceran,Ceriodaphnia dubia

S,U Copper 

sulfate

428 EC50 36.4 

Cladoceran,Ceriodaphnia dubia

S,U Copper 

sulfate

410 EC50 11.7 

Cladoceran,Ceriodaphnia dubia

S,U Copper 

sulfate

494 EC50 12.3 

Cladoceran,Ceriodaphnia dubia

S,U Copper 

sulfate

440 EC50 12 

Cladoceran, Ceriodaphnia dubia

S,U Copper 

chloride

90 1 hr NOEC

(ingestion)

30 - 

Cladoceran (<24 hr),Ceriodaphnia dubia

S,M,D Copper 

sulfate

6-10 48 hr LC50 - 2.72 

Cladoceran (<12 hr),Ceriodaphnia dubia

S,M,D - 113.6 48 hr LC50 - 52 

Cladoceran (<12 hr),Ceriodaphnia dubia

S,M,D - 113.6 48 hr LC50 - 76 

Cladoceran (<12 hr),Ceriodaphnia dubia

S,M,D - 113.6 48 hr LC50 - 91 

Cladoceran (<48 h),Ceriodaphnia dubia

S,M,T Copper 

nitrate

280 - 300 48 hr LC50 9.5 - 

Cladoceran (<48 h),Ceriodaphnia dubia

S,M,T Copper 

nitrate

280 - 300 48 hr LC50 28 - 

Cladoceran (<48 h),Ceriodaphnia dubia

S,M,T Copper 

nitrate

280 - 300 48 hr LC50 200 - 

Cladoceran (<24 hr),Ceriodaphnia dubia

S,M,T,D Copper 

nitrate

100 48 hr LC50 66 60.72 

Cladoceran, Ceriodaphnia dubia

R,U Copper 

nitrate

111 10 days LC50 53 - 

Cladoceran, Ceriodaphnia dubia

R,U Copper 

nitrate

111 10 days NOEC 

(reproduction)

96 - 

Cladoceran, Ceriodaphnia dubia

R,U Copper 

sulfate

90 - LOEC

(reproduction)

44 - 

Cladoceran, Ceriodaphnia dubia

R,U Copper 

sulfate

90 - LOEC

(reproduction)

40 - 

Cladoceran, Ceriodaphnia dubia

R,M,T - 20 - IC50 

(reproduction) 

5 - 

Cladoceran (<24 hrs),Ceriodaphnia reticulata

S, U Copper 

chloride

240 EC50 23 

Cladoceran,Ceriodubia reticulata

S,U - 43-45 EC50 17 

Cladoceran,Daphnia magna

- Copper 

sulfate

- 72 hr EC50

(mobility; 100 C)

61 - 

B-9

Appendix B. Other Data on Effects of Copper on Freshwater Organisms
cies Methoda Chemical

Hardness 

(mg/L as 

CaCO3)

Duration Effect

Total

Concentration

(μg/L)b

Dissolved

Concentration

(μg/L)

Cladoceran,Daphnia magna

- Copper 

sulfate

- 72 hr EC50

(mobility; 150 C)

70 - 

Cladoceran,Daphnia magna

- Copper 

sulfate

- 72 hr EC50

(mobility; 200 C)

21 - 

Cladoceran,Daphnia magna

- Copper 

sulfate

- 72 hr EC50

(mobility; 300 C)

9.3 - 

Cladoceran,Daphnia magna

S,U Copper 

sulfate

- 16 hr EC 50

(mobility)

38 - 

Cladoceran (<8 hr),Daphnia magna

S,U Copper 

chloride

- 64 hr Immobilization threshold 12.7 - 

Cladoceran (1 mm),Daphnia magna

S,U Copper 

nitrate

100 24 hr EC 50

(mobility)

50 - 

Cladoceran (1 mm),Daphnia magna

S,U Copper 

nitrate

200 24 hr EC 50

(mobility)

70 - 

Cladoceran,Daphnia magna

S,U - 100 48 hr EC50

(mobility)

254 - 

Cladoceran,Daphnia magna

S,U - 100 49 hr EC50

(mobility)

1,239 - 

Cladoceran,Daphnia magna

S,U Copper 

sulfate

45 48 hr EC50

(mobility; 50 C)

90 - 

Cladoceran,Daphnia magna

S,U Copper 

sulfate

45 48 hr EC50

(mobility; 100 C)

70 - 

Cladoceran,Daphnia magna

S,U Copper 

sulfate

45 48 hr EC50

(mobility; 150 C)

40 - 

Cladoceran,Daphnia magna

S,U Copper 

sulfate

45 48 hr EC50

(mobility; 250 C)

7 - 

Cladoceran (4 days),Daphnia magna

S,U Copper 

sulfate

- 24 hr EC50

(filtration rate)

59 - 

Cladoceran (24-48 hr),Daphnia magna

S,U Copper 

sulfate

90 24 hr EC50

(mobility)

380 - 

Cladoceran,Daphnia magna

S,U Copper 

sulfate

50 EC50 7 

Cladoceran,Daphnia magna

S,U Copper 

sulfate

- 48 hr EC50

(mobility)

45 - 

Cladoceran (<24 hr),Daphnia magna

S,U Copper 

sulfate

145 Life span 

(ca. 18 wk)

Chronic limits 

(inst. rate of population growth)

70 - 

Cladoceran (<24 hrs),Daphnia magna

S,M,D Copper 

sulfate

72-80 48 hr LC50 - 11.3 

Cladoceran (<24 hrs),Daphnia magna

S,M,I - 180 - LC50 55.3 - 

Cladoceran (<24 hr),Daphnia magna

S,M,I Copper 

sulfate

100 48 hr EC50

(mobility)

46.0 - 

Cladoceran (<24 hr),Daphnia magna

S,M,I Copper 

sulfate

100 48 hr EC50

(mobility)

57.2 - 

B-10

Appendix B. Other Data on Effects of Copper on Freshwater Organisms
Species Methoda Chemical

Hardness 

(mg/L as 

CaCO3)

Duration Effect

Total

Concentration

(μg/L)b

Dissolved

Concentration

(μg/L)

Cladoceran (<24 hr),Daphnia magna

S,M,I Copper 

sulfate

100 48 hr EC50

(mobility)

67.8 - 

Cladoceran (<24 hr),Daphnia magna

S,M,T Copper 

sulfate

100 72 hr EC50

(mobility)

52.8 - 

Cladoceran (<24 hr),Daphnia magna

S,M,T Copper 

sulfate

100 72 hr EC50

(mobility)

56.3 - 

Cladoceran (<24 hr),Daphnia magna

S,M,T Copper 

chloride

85 96 hr EC50

(mobility)

130 - 

Cladoceran (24 hr),Daphnia magna

R,U Copper 

sulfate

- 48 hr EC50

(mobility)

18 - 

Cladoceran (<24 hr),Daphnia parvula

S,U Copper 

sulfate

145 72 hr EC50

(mobility)

72 - 

Cladoceran (<24 hr),Daphnia parvula

S,U Copper 

sulfate

145 72 hr EC50

(mobility)

57 - 

Cladoceran (<24 hr),Daphnia parvula

S,U Copper 

sulfate

145 Life span 

(ca. 10 wk)

Chronic limits (inst. rate of population 

growth)

50 - 

Cladoceran,Daphnia pulex

S,U Copper 

sulfate

45 EC50 10 

Cladoceran,Daphnia pulex

S,U - 45 EC50 53 

Cladoceran (<24 hrs),Daphnia pulex

S, U Copper 

chloride

240 EC50 31 

Cladoceran (<24 hrs),Daphnia pulex

S, U Copper 

sulfate

33.8 EC50 3.6 

Cladoceran (<24 hrs),Daphnia pulex

S,U Copper 

chloride

80-90 EC50 18 

Cladoceran (<24 hrs),Daphnia pulex

S,U Copper 

chloride

80-90 EC50 24 

Cladoceran (<24 hrs),Daphnia pulex

S,U Copper 

chloride

80-90 EC50 22 

Cladoceran (<24 hr),Daphnia pulex

S,U Copper 

sulfate

145 72 hr EC50

(mobility)

86 - 

Cladoceran (<24 hr),Daphnia pulex

S,U Copper 

sulfate

145 72 hr EC50

(mobility)

54 - 

Cladoceran (<24 hr),Daphnia pulex

S,U Copper 

sulfate

145 Life span 

(ca. 7 wk)

Chronic limits (inst. rate of population 

growth)

50 - 

Cladoceran,Daphnia pulex

S,U Copper 

sulfate

45 48 hr EC50

(mobility)

70 - 

Cladoceran,Daphnia pulex

S,U Copper 

sulfate

45 48 hr EC50

(mobility)

60 - 

Cladoceran,Daphnia pulex

S,U Copper 

sulfate

45 48 hr EC50

(mobility)

20 - 

Cladoceran,Daphnia pulex

S,U Copper 

sulfate

45 48 hr EC50

(mobility)

56 - 

B-11

Appendix B. Other Data on Effects of Copper on Freshwater Organisms
Species Methoda Chemical

Hardness 

(mg/L as 

CaCO3)

Duration Effect

Total

Concentration

(μg/L)b

Dissolved

Concentration

(μg/L)

Cladoceran (<24 hr),Daphnia pulex

S,U Copper 

sulfate

200 24 hr EC50

(mobility)

37.5 - 

Cladoceran,Daphnia pulex

S,M,T Copper 

sulfate

106 48 hr EC50

(mobility)

29 - 

Cladoceran,Daphnia pulex

S,M,T Copper 

sulfate

106 48 hr EC50

(mobility)

20 - 

Cladoceran,Daphnia pulex

S,M,T Copper 

sulfate

106 48 hr EC50

(mobility)

25 - 

Cladoceran,Daphnia pulex

R,U Copper 

sulfate

85 21 days Reduced fecundity 3 - 

Cladoceran,Daphnia pulex

R,M,T Copper 

sulfate

106 70 days Significantly shortened life span; 

reduced brood size

20 - 

Cladoceran,Daphnia pulicaria

S,M,T - 31 48 hr EC50

(mobility; TOC=14 mg/L)

55.4 - 

Cladoceran,Daphnia pulicaria

S,M,T - 29 49 hr EC50

(mobility; TOC=13 mg/L)

55.3 - 

Cladoceran,Daphnia pulicaria

S,M,T - 28 50 hr EC50

(mobility; TOC=13 mg/L)

53.3 - 

Cladoceran,Daphnia pulicaria

S,M,T - 28 50 hr EC50

(mobility; TOC=28 mg/L)

97.2 - 

Cladoceran,Daphnia pulicaria

S,M,T - 100 51 hr EC50

(mobility; TOC=34 mg/L)

199 - 

Cladoceran,Daphnia pulicaria

S,M,T - 86 52 hr EC50

(mobility; TOC=34 mg/L)

627 - 

Cladoceran,Daphnia pulicaria

S,M,T - 84 53 hr EC50

(mobility; TOC=32 mg/L)

165 - 

Cladoceran,Daphnia pulicaria

S,M,T - 16 54 hr EC50

(mobility; TOC=12 mg/L)

35.5 - 

Cladoceran,Daphnia pulicaria

S,M,T - 151 55 hr EC50

(mobility; TOC=13 mg/L)

78.8 - 

Cladoceran,Daphnia pulicaria

S,M,T - 96 56 hr EC50

(mobility; TOC=28 mg/L)

113 - 

Cladoceran,Daphnia pulicaria

S,M,T - 26 57 hr EC50

(mobility; TOC=25 mg/L)

76.4 - 

Cladoceran,Daphnia pulicaria

S,M,T - 84 58 hr EC50

(mobility; TOC=13 mg/L)

84.7 - 

Cladoceran,Daphnia pulicaria

S,M,T - 92 59 hr EC50

(mobility; TOC=21 mg/L)

184 - 

Cladoceran,Daphnia pulicaria

S,M,T - 106 60 hr EC50

(mobility; TOC=34 mg/L)

240 - 

Cladoceran,Daphnia pulicaria

S,M,T Copper 

sulfate

106 48 hr LC50 240 - 

Cladoceran,Simocephalus serrulatus

S,M,T Copper 

nitrate

8 24 hr EC50

(mobility; TOC=11 mg/L)

12 - 

B-12

Appendix B. Other Data on Effects of Copper on Freshwater Organisms
Species Methoda Chemical

Hardness 

(mg/L as 

CaCO3)

Duration Effect

Total

Concentration

(μg/L)b

Dissolved

Concentration

(μg/L)

Cladoceran,Simocephalus serrulatus

S,M,T Copper 

nitrate

16 25 hr EC50

(mobility; TOC=12.4 mg/L)

7.2 - 

Cladoceran,Simocephalus serrulatus

S,M,T Copper 

nitrate

16 26 hr EC50

(mobility; TOC=15.6 mg/L)

24.5 - 

Cladoceran (<24 hr),Simocephalus vetulus

S,U - 45 57 

Cladoceran (life cycle),Bosmina longirostris

R,U Copper 

sulfate

- 13 days LOEC

(intrinsic rate of population increase)

18 - 

Copepods (mixed sp),Primarily Acanthocyclops vernalis and Diacyclops thomasi

R,M,I Copper 

chloride

- 1 wk EC20

(growth)

42 - 

Copepod (adults and copepodids

V),Tropocyclops prasinus mexicanus

S, U Copper 

sulfate

10 29 

Copepod (adults and copepodidsV), Tropocyclops prasinusmexicanus

S, U Copper 

sulfate

10 96 hr LC50 247 - 

Amphipod (0.4 cm),Crangonyx pseudogracilis

R,U Copper 

sulfate

45-55 1290 

Amphipod (4 mm),Crangonyx psuedogracilis

R,U Copper 

sulfate

50 48 hr LC50 2,440 - 

Amphipod,Gammarus fasciatus

S,U Copper 

sulfate

206 48 hr LC50 210 - 

Amphipod,Gammarus lacustris

S,U Copper 

sulfate

- 96 hr LC50 1,500 - 

Amphipod (2-3 wk),Hyallela azteca

S,M,T Copper 

sulfate

6-10 - LC50 65.6 - 

Amphipod (0-1 wk),Hyallela azteca

R,M,T Copper 

nitrate

130 10 wk Significant mortality 25.4 - 

Amphipod (7-14 days),Hyallela azteca

F,M,T Copper 

sulfate

46 10 days LC50 31 - 

Crayfish (intermoult adult, 19.6 g),Cambarus robustus

S,M,D - 10-12 96 hr LC50 - 830 

Crayfish (1.9-3.2 cm),Orconectes limosus

S,M,T Copper 

chloride

- 96 hr LC50 600 - 

Crayfish (3.0-3.5 cm),Orconectes rusticus

F,U Copper 

sulfate

100-125 3,000 

Crayfish (embryo),Orconectes rusticus

F,U Copper 

sulfate

113 2 wk 52% mortality of newly 

hatched young

250 - 

B-13

Appendix B. Other Data on Effects of Copper on Freshwater Organisms
Species Methoda Chemical

Hardness 

(mg/L as 

CaCO3)

Duration Effect

Total

Concentration

(μg/L)b

Dissolved

Concentration

(μg/L)

Crayfish (3.14 mg dry wt.),Orconectes rusticus

F,U Copper 

sulfate

113 2 wk 23% reduction in growth 15 - 

Crayfish (30-40 mm),Orconectes sp.

 - 113 48 hr LC50 2,370 - 

Crayfish,Procambarus clarkii

F,M,T Copper 

chloride

17 1358 hr LC50 657 - 

Mayfly (6th-8th instar),Stenonema sp.

S,M,T - 110 48 hr LC50 453 - 

Mayfly,Cloeon dipterium

- Copper 

sulfate

- 72 hr LC50 

(100 C)

193 - 

Mayfly,Cloeon dipterium

- - - 72 hr LC50 

(150 C)

95.2 - 

Mayfly,Cloeon dipterium

- - - 72 hr LC50 

(250 C)

53 - 

Mayfly,Cloeon dipterium

- - - 72 hr LC50 

(300 C)

4.8 - 

Mayfly,Ephemerella grandis

F,M,T Copper 

sulfate

50 14 days LC50 180-200 - 

Mayfly,Ephemerella subvaria

S,M Copper 

sulfate

44 48 hr LC50 320 - 

Mayfly (6th-8th instar),Isonychia bicolor

S,M,T - 110 48 hr LC50 223 - 

Stonefly,Pteronarcys californica

F,M,T Copper 

sulfate

50 14 days LC50 12,000 - 

Caddisfly,Hydropsyche betteni

S,M,T Copper 

sulfate

44 14 days LC50 32,000 - 

Midge (2nd instar),Chironomus riparius

S,M,T - 110 48 hr LC50 1,170 - 

Midge (1st instar),Chironomus tentans

S,U Copper 

sulfate

42.7 16.7 

Midge (1st instar),Chironomus tentans

S,U Copper 

sulfate

109.6 36.5 

Midge (1st instar),Chironomus tentans

S,U Copper 

sulfate

172.3 98.2 

Midge (4th instar),Chironomus tentans

S,U Copper 

sulfate

42.7 211 

Midge (4th instar),Chironomus tentans

S,U Copper 

sulfate

109.6 977 

Midge (4th instar),Chironomus tentans

S,U Copper 

sulfate

172.3 1184 

Midge,Chironomus tentans

S,U Copper 

sulfate

25 327 

Midge (2nd instar),Chironomus tentans

S,M,T Copper 

sulfate

8 96 hr LC50 630 - 

B-14

Appendix B. Other Data on Effects of Copper on Freshwater Organisms
Species Methoda Chemical

Hardness 

(mg/L as 

CaCO3)

Duration Effect

Total

Concentration

(μg/L)b

Dissolved

Concentration

(μg/L)

Midge (4th instar),Chironomus tentans

F,M,T Copper 

chloride

36 20 days LC50 77.5 - 

Midge (embryo),Tanytarsus dissimilis

S,M,T Copper 

chloride

46.8 10 days LC50 16.3 - 

Midge,Unidentified

F,M,T,D Copper 

sulfate

200 32 wk Emergence 30 - 

Bryozoan (2-3 day ancestrula),Lophopodella carteri

S,U - 190-220 510 

Bryozoan (2-3 day ancestrula),Pectinatella magnifica

S,U - 190-220 140 

Bryozoan (2-3 day ancestrula),Plumatella emarginata

S,U - 190-220 140 

American eel (5.5 cm glass eel stage),Anguilla rostrata

S,U Copper 

sulfate

40-48 96 hr LC50 2,540 

American eel (9.7 cm black eel stage),Anguilla rostrata

S,U Copper 

sulfate

40-48 96 hr LC50 3,200 

American eel,Anguilla rostrata

S,M,T Copper 

nitrate

53 96 hr LC50 6,400 - 

American eel,Anguilla rostrata

S,M,T Copper 

nitrate

55 96 hr LC50 6,000 - 

Arctic grayling (larva),Thymallus arcticus

S,U Copper 

sulfate

41.3 96 hr LC50 67.5 

Arctic grayling (larva),Thymallus arcticus

S,U Copper 

sulfate

41.3 96 hr LC50 23.9 

Arctic grayling (larva),Thymallus arcticus

S,U Copper 

sulfate

41.3 96 hr LC50 131 

Arctic grayling (swim-up),Thymallus arcticus

S,U Copper 

sulfate

41.3 96 hr LC50 9.6 

Arctic grayling (0.20 g juvenile),Thymallus arcticus

S,U Copper 

sulfate

41.3 96 hr LC50 2.7 

Arctic grayling (0.34 g juvenile),Thymallus arcticus

S,U Copper 

sulfate

41.3 96 hr LC50 2.58 

Arctic grayling (0.81 g juvenile),Thymallus arcticus

S,U Copper 

sulfate

41.3 96 hr LC50 49.3 

Arctic grayling (0.85 g juvenile),Thymallus arcticus

S,U Copper 

sulfate

41.3 96 hr LC50 30 

Coho salmon (larva),Oncorhynchus kisutch

S,U Copper 

sulfate

41.3 96 hr LC50 21 

Coho salmon (larva),Oncorhynchus kisutch

S,U Copper 

sulfate

41.3 96 hr LC50 19.3 

Coho salmon (0.41 g juvenile),Oncorhynchus kisutch

S,U Copper 

sulfate

41.3 96 hr LC50 15.1 

B-15

Appendix B. Other Data on Effects of Copper on Freshwater Organisms
Species Methoda Chemical

Hardness 

(mg/L as 

CaCO3)

Duration Effect

Total

Concentration

(μg/L)b

Dissolved

Concentration

(μg/L)

Coho salmon (0.47 g juvenile),Oncorhynchus kisutch

S,U Copper 

sulfate

41.3 96 hr LC50 23.9 

Coho salmon (0.87 g juvenile),Oncorhynchus kisutch

S,U Copper 

sulfate

41.3 96 hr LC50 31.9 

Coho salmon (10 cm),Oncorhynchus kisutch

S,U Copper 

sulfate

- 72 hr LC50 280 - 

Coho salmon (9.7 cm),Oncorhynchus kisutch

S,U Copper 

sulfate

- 72 hr LC50 190 - 

Coho salmon (9.7 cm),Oncorhynchus kisutch

S,U Copper 

sulfate

- 72 hr LC50 480 - 

Coho salmon (juvenile),Oncorhynchus kisutch

R,M,T,I - 33 96 hr LC50 

(TOC=7.3 mg/L)

164 - 

Coho salmon (juvenile),Oncorhynchus kisutch

R,M,T,I - 33 96 hr LC50 286 

Coho salmon (6.3 cm),Oncorhynchus kisutch

F,U Copper 

sulfate

- 30 days LC50 360 - 

Coho salmon (6.3 cm),Oncorhynchus kisutch

F,U Copper 

sulfate

- 72 hr LC50 370 - 

Coho salmon (smolts),Oncorhynchus kisutch

F,M,T Copper 

chloride

91 144 hr Decrease in survival upon transfer to 

30 ppt seawater

20 - 

Coho salmon (smolts >10 cm),Oncorhynchus kisutch

F,M,T Copper 

chloride

91 165 days Decrease in downstream migration 

after release

5 - 

Coho salmon (7.8 cm),Oncorhynchus kisutch

F,M,T Copper 

acetate

276 14 wk 15% reduction in growth 70 - 

Coho salmon (7.8 cm),Oncorhynchus kisutch

- - 276 7 days LC50 220 - 

Coho salmon (3-8 g),Oncorhynchus kisutch

F,M,T Copper 

acetate

280 7 days LC50 275 - 

Coho salmon (3-8 g),Oncorhynchus kisutch

F,M,T Copper 

acetate

280 7 days LC50 (acclimated to copper for 2 wk) 383 - 

Coho salmon (parr),Oncorhynchus kisutch

F,M,T,D,I - 24.4 61 days NOEC

(growth and survival)

22 - 

Coho salmon,Oncorhynchus kisutch

F,M,T,D,I - 31.1 60 days NOEC

(growth and survival)

18 - 

Coho salmon (parr),Oncorhynchus kisutch

F,M,T,D,I - 31 61 days NOEC

(growth and survival)

33 - 

Rainbow trout (15-40g) Oncorhynchus mykiss

F,M, Copper 

chloride

-- 120 hr LA50 (50% mortality) ~1.4 ug Cu/g gill - 

Sockeye salmon (yeasrling),Oncorhynchus nerka

S,U Copper 

sulfate

12 1-24 hr Drastic increase in plasma 

corticosteroids

64 - 

Sockeye salmon (fry, 0.132 g, 2.95 cm),Oncorhynchus nerka

R,M,T Copper 

chloride

36-46 96 hr LC50 220 - 

B-16

Appendix B. Other Data on Effects of Copper on Freshwater Organisms
Species Methoda Chemical

Hardness 

(mg/L as 

CaCO3)

Duration Effect

Total

Concentration

(μg/L)b

Dissolved

Concentration

(μg/L)

Sockeye salmon (fry, 0.132 g, 2.95 cm),Oncorhynchus nerka

R,M,T Copper 

chloride

36-46 96 hr LC50 210 - 

Sockeye salmon (fry, 0.132 g, 2.95 cm),Oncorhynchus nerka

R,M,T Copper 

chloride

36-46 96 hr LC50 240 - 

Sockeye salmon (fry, 0.132 g, 2.95 cm),Oncorhynchus nerka

R,M,T Copper 

chloride

36-46 96 hr LC50 103 - 

Sockeye salmon (fry, 0.132 g, 2.95 cm),Oncorhynchus nerka

R,M,T Copper 

chloride

36-46 96 hr LC50 240 - 

Chinook salmon (18-21 weeks),Oncorhynchus tshawytscha

S,U Copper 

sulfate

211 96 hr LC50 58 

Chinook salmon (18-21 weeks),Oncorhynchus tshawytscha

S,U Copper 

sulfate

211 96 hr LC50 54 

Chinook salmon (18-21 weeks),Oncorhynchus tshawytscha

S,U Copper 

sulfate

343 96 hr LC50 60 

Chinook salmon (5.2 cm),Oncorhynchus tshawytscha

S,U Copper 

nitrate

- 5 days LC50 178 - 

Chinook salmon (eyed embryos),Oncorhynchus tshawytscha

F,M,D Copper 

sulfate

44 26 days 93% mortality 41.67 - 

Chinook salmon (alevin),Oncorhynchus tshawytscha

F,M,T Copper 

chloride

23 200 hr LC50 20 - 

Chinook salmon (alevin),Oncorhynchus tshawytscha

F,M,T Copper 

chloride

23 200 hr LC10 15 - 

Chinook salmon (swimup),Oncorhynchus tshawytscha

F,M,T Copper 

chloride

23 200 hr LC50 19 - 

Chinook salmon (swimup),Oncorhynchus tshawytscha

F,M,T Copper 

chloride

23 200 hr LC10 14 - 

Chinook salmon (parr),Oncorhynchus tshawytscha

F,M,T Copper 

chloride

23 200 hr LC50 30 - 

Chinook salmon (parr),Oncorhynchus tshawytscha

F,M,T Copper 

chloride

23 200 hr LC10 17 - 

Chinook salmon (smolt),Oncorhynchus tshawytscha

F,M,T Copper 

chloride

23 200 hr LC50 26 - 

Chinook salmon (smolt),Oncorhynchus tshawytscha

F,M,T Copper 

chloride

23 200 hr LC10 18 - 

Chinook salmon (3.9-6.8 cm),Oncorhynchus tshawytscha

F,M,T Copper 

sulfate

20-22 96 hr LC50 32 - 

Cutthroat trout (3-5 mo),Oncorhynchus clarki

F,M Copper 

chloride

50 20 min avoidance of copper 7.708 - 

B-17

Appendix B. Other Data on Effects of Copper on Freshwater Organisms
Species Methoda Chemical

Hardness 

(mg/L as 

CaCO3)

Duration Effect

Total

Concentration

(μg/L)b

Dissolved

Concentration

(μg/L)

Rainbow trout,Oncorhynchus mykiss

- - 320 48 hr LC50 500 - 

Rainbow trout (9-16 cm),Oncorhynchus mykiss

In situ - 21-26 48 hr LC50 70 - 

Rainbow trout (0.4 g),Oncorhynchus mykiss

S,U Copper 

sulfate

- 96 hr LC50 185 - 

Rainbow trout (larva),Oncorhynchus mykiss

S, U Copper 

sulfate

41.3 96 hr LC50 36 

Rainbow trout (0.60 g juvenile),Oncorhynchus mykiss

S, U Copper 

sulfate

41.3 96 hr LC50 13.8 

Rainbow trout (13-15 cm),Oncorhynchus mykiss

S,U Copper 

sulfate

250 72 hr LC50 580 - 

Rainbow trout (13-15 cm),Oncorhynchus mykiss

S,U Copper 

sulfate

250 72 hr LC50 960 - 

Rainbow trout (3.2 cm),Oncorhynchus mykiss

S,U Copper 

sulfate

- 24 hr LC50 140 - 

Rainbow trout (3.2 cm),Oncorhynchus mykiss

S,U Copper 

sulfate

- 24 hr LC50 130 - 

Rainbow trout (4.0-10.6 cm),Oncorhynchus mykiss

S,U Copper 

sulfate

45 24 hr LC50 

(50 C)

950 - 

Rainbow trout (4.0-10.6 cm),Oncorhynchus mykiss

S,U Copper 

sulfate

45 24 hr LC50 

(150 C) 

430 - 

Rainbow trout (4.0-10.6 cm),Oncorhynchus mykiss

S,U Copper 

sulfate

45 24 hr LC50 

(300 C)

150 - 

Rainbow trout (0.52-1.55 g), Oncorhynchus mykiss

S,U Copper 

sulfate

- 96 hr LC50 

(Silver Cup diet)

23.9 - 

Rainbow trout (0.41-2.03 g), Oncorhynchus mykiss

S,U Copper 

sulfate

- 96 hr LC50 

(purified H440)

11.3 - 

Rainbow trout (0.0.40-1.68 g), Oncorhynchus mykiss

S,U Copper 

sulfate

- 96 hr LC50 

(SD-9 diet)

15.9 - 

Rainbow trout (0.0.34-1.52 g), Oncorhynchus mykiss

S,U Copper 

sulfate

- 96 hr LC50 

(liver diet)

14.3 - 

Rainbow trout (0.0.38-1.30 g), Oncorhynchus mykiss

S,U Copper 

sulfate

- 96 hr LC50 

(brine shrimp diet)

11.3 - 

Rainbow trout (embryo),Oncorhynchus mykiss

S,U Copper 

chloride

30 56 hr LC50 100 - 

Rainbow trout (6.6 cm),Oncorhynchus mykiss

R,U Copper 

sulfate

320 72 hr LC50 1,100 - 

Rainbow trout (6.6 cm),Oncorhynchus mykiss

R,U Copper 

sulfate

17.5 7 days LC50 44 - 

Rainbow trout,Oncorhynchus mykiss

R,U Copper 

sulfate

320 48 hr LC50 270 - 

Rainbow trout (yearling),Oncorhynchus mykiss

R,U Copper 

sulfate

240 48 hr LC50 750 - 

B-18

Appendix B. Other Data on Effects of Copper on Freshwater Organisms
Species Methoda Chemical

Hardness 

(mg/L as 

CaCO3)

Duration Effect

Total

Concentration

(μg/L)b

Dissolved

Concentration

(μg/L)

Rainbow trout (13-15 cm),Oncorhynchus mykiss

R,U Copper 

sulfate

250 8 days LC50 500 - 

Rainbow trout (embryo),Oncorhynchus mykiss

R,U Copper 

sulfate

104 28 days LC50 90 - 

Rainbow trout (embryo),Oncorhynchus mykiss

R,U Copper 

sulfate

101 28 days EC50

(death or deformity)

110 - 

Rainbow trout (embryo),Oncorhynchus mykiss

R,U Copper 

sulfate

101 28 days EC10

(death or deformity)

16.5 - 

Rainbow trout (eyed embryos),Oncorhynchus mykiss

R,U Copper 

sulfate

- 96 hr LC50 1,150 - 

Rainbow trout (larva),Oncorhynchus mykiss

R,U Copper 

sulfate

- 96 hr LC50 430 - 

Rainbow trout (16-18 cm),Oncorhynchus mykiss

R,U Copper 

sulfate

- 96 hr LC50 930 - 

Rainbow trout (embryo),Oncorhynchus mykiss

R,M,T Copper 

sulfate

62.9 7-9 mo Lesions in olfactory rosettes 22 - 

Rainbow trout (embryo),Oncorhynchus mykiss

R,M,T Copper 

sulfate

62.9 7-9 mo 31% mortality 22 - 

Rainbow trout (eyed embryos),Oncorhynchus mykiss

R,M,T Copper 

sulfate

40-48 96 hr LC50 400 - 

Rainbow trout (yearling),Oncorhynchus mykiss

R,M,T Copper 

sulfate

36.5 21 days Elevated plasma cortisol returned 

to normal

45 - 

Rainbow trout (embryo),Oncorhynchus mykiss

R,M,T Copper 

sulfate

44 96 hr 15-20% post-hatch mortality 80 - 

Rainbow trout (embryo),Oncorhynchus mykiss

R,M,T Copper 

sulfate

62.9 7-9 mo Inhibited olfactory discrimination 22 - 

Rainbow trout (5.1-7.6 cm),Oncorhynchus mykiss

F,U Copper 

nitrate

- 96 hr LC50 253 - 

Rainbow trout (11 cm),Oncorhynchus mykiss

F,U - 100 96 hr LC50 250 - 

Rainbow trout (5 wk post swimup)Oncorhynchus mykiss

F,U Copper 

sulfate

89.5 1 hr Avoidance 10 - 

Rainbow trout (18.5-26.5 cm),Oncorhynchus mykiss

F,U Copper 

sulfate

90 2 hr 55% depressed olfactory response 50 - 

Rainbow trout (3.2 cm),Oncorhynchus mykiss

F,M,I Copper 

sulfate

- 8 days LC50 500 - 

Rainbow trout (12-16 cm),Oncorhynchus mykiss

F,M,T Copper 

sulfate

300 14 days LC50 870 - 

Rainbow trout (adult),Oncorhynchus mykiss

F,M,T Copper 

chloride

42 - LC50 57 - 

Rainbow trout (53.5 g),Oncorhynchus mykiss

F,M,T Copper 

sulfate

365 96 hr LC50 465 - 

B-19

Appendix B. Other Data on Effects of Copper on Freshwater Organisms
Species Methoda Chemical

Hardness 

(mg/L as 

CaCO3)

Duration Effect

Total

Concentration

(μg/L)b

Dissolved

Concentration

(μg/L)

Rainbow trout (53.5 g),Oncorhynchus mykiss

F,M,T Copper 

sulfate

365 15 days Transient decrease in food 

consumption

100 - 

Rainbow trout (alevin),Oncorhynchus mykiss

F,M,T Copper 

chloride

24 200 hr LC50 20 - 

Rainbow trout (alevin),Oncorhynchus mykiss

F,M,T Copper 

chloride

24 200 hr LC10 19 - 

Rainbow trout (swimup),Oncorhynchus mykiss

F,M,T Copper 

chloride

24 200 hr LC50 17 - 

Rainbow trout (swimup),Oncorhynchus mykiss

F,M,T Copper 

chloride

24 200 hr LC10 9 - 

Rainbow trout (parr),Oncorhynchus mykiss

F,M,T Copper 

chloride

25 200 hr LC50 15 - 

Rainbow trout (parr),Oncorhynchus mykiss

F,M,T Copper 

chloride

25 200 hr LC10 8 - 

Rainbow trout (smolt),Oncorhynchus mykiss

F,M,T Copper 

chloride

25 200 hr LC50 21 - 

Rainbow trout (smolt),Oncorhynchus mykiss

F,M,T Copper 

chloride

25 200 hr LC10 7 - 

Rainbow trout,Oncorhynchus mykiss

F,M,T Copper 

sulfate

112.4 80 min Avoidance threshold 74 - 

Rainbow trout (>8 g),Oncorhynchus mykiss

F,M,T Copper 

sulfate

49 15-18 days LC50 48 - 

Rainbow trout (>8 g),Oncorhynchus mykiss

F,M,T Copper 

sulfate

51 15-18 days LC50 46 - 

Rainbow trout (>8 g),Oncorhynchus mykiss

F,M,T Copper 

sulfate

57 15-18 days LC50 63 - 

Rainbow trout (>8 g),Oncorhynchus mykiss

F,M,T Copper 

sulfate

12 15-18 days LC50 19 - 

Rainbow trout (>8 g),Oncorhynchus mykiss

F,M,T Copper 

sulfate

99 15-18 days LC50 54 - 

Rainbow trout (>8 g),Oncorhynchus mykiss

F,M,T Copper 

sulfate

98 15-18 days LC50 78 - 

Rainbow trout (>8 g),Oncorhynchus mykiss

F,M,T Copper 

sulfate

12 15-18 days LC50 18 - 

Rainbow trout (>8 g),Oncorhynchus mykiss

F,M,T Copper 

sulfate

97 15-18 days LC50 96 - 

Rainbow trout (200-250 g),Oncorhynchus mykiss

F,M,T Copper 

sulfate

320 4 mo Altered liver and blood enzymes and 

mitochondrial function

30 - 

Rainbow trout (7 cm),Oncorhynchus mykiss

F,M,T Copper 

chloride

28.4 20 min Avoidance 6.4 - 

Rainbow trout (2.70 g),Oncorhynchus mykiss

F,M,T Copper 

chloride

9.2 96 hr LC50 4.2 - 

Rainbow trout (2.88 g),Oncorhynchus mykiss

F,M,T Copper 

chloride

9.2 96 hr LC50 66 - 

B-20

Appendix B. Other Data on Effects of Copper on Freshwater Organisms
Species Methoda Chemical

Hardness 

(mg/L as 

CaCO3)

Duration Effect

Total

Concentration

(μg/L)b

Dissolved

Concentration

(μg/L)

Rainbow trout (2.88 g),Oncorhynchus mykiss

F,M,T Copper 

chloride

9.2 168 hr LC50 36.7 - 

Rainbow trout (2.70 g),Oncorhynchus mykiss

F,M,T Copper 

chloride

9.2 168 hr LC50 3.1 - 

Rainbow trout (2.65 g),Oncorhynchus mykiss

F,M,T Copper 

sulfate

9.2 168 hr LC50 2.3 - 

Rainbow trout (5 day embryo),Oncorhynchus mykiss

F,M,T Copper 

sulfate

87.7 48 hr LC50 8,000 - 

Rainbow trout (10 day embryo),Oncorhynchus mykiss

F,M,T Copper 

sulfate

87.7 48 hr LC50 2,000 - 

Rainbow trout (15 day embryo),Oncorhynchus mykiss

F,M,T Copper 

sulfate

87.7 48 hr LC50 400 - 

Rainbow trout (22 day embryo),Oncorhynchus mykiss

F,M,T Copper 

sulfate

87.7 48 hr LC50 600 - 

Rainbow trout (29 day embryo),Oncorhynchus mykiss

F,M,T Copper 

sulfate

87.7 48 hr LC50 400 - 

Rainbow trout (36 day embryo),Oncorhynchus mykiss

F,M,T Copper 

sulfate

87.7 48 hr LC50 100 - 

Rainbow trout (2 day larva),Oncorhynchus mykiss

F,M,T Copper 

sulfate

87.7 48 hr LC50 100 - 

Rainbow trout (7 day larva),Oncorhynchus mykiss

F,M,T Copper 

nitrate

87.7 48 hr LC50 100 - 

Rainbow trout (yearling),Oncorhynchus mykiss

F,M,T Copper 

sulfate

63 15 days Olfactory receptor degeneration 20 - 

Rainbow trout (swimup),Oncorhynchus mykiss

F,M,T Copper 

sulfate

60.9 13-40 wk Inhibited olfactory discrimination 20 - 

Rainbow trout (swimup),Oncorhynchus mykiss

F,M,T Copper 

sulfate

60.9 40 wk 43% mortality 40 - 

Rainbow trout (9.0-11.5 cm, 10.6 g),Oncorhynchus mykiss

F,M,T Copper 

sulfate

284 96 hr LC50 650 - 

Rainbow trout (3.5 cm),Oncorhynchus mykiss

F,M,T Copper 

chloride

24.2 96 hr LC50 12.7 - 

Rainbow trout (3.5 cm),Oncorhynchus mykiss

F,M,T Copper 

chloride

24.2 96 hr LC50 16.6 - 

Rainbow trout (3.5 cm),Oncorhynchus mykiss

F,M,T Copper 

chloride

24.2 96 hr LC50 21.4 - 

Rainbow trout (3.5 cm),Oncorhynchus mykiss

F,M,T Copper 

chloride

24.2 96 hr LC50 34.2 - 

Rainbow trout (10.0 g),Oncorhynchus mykiss

F,M,D Copper 

sulfate

362 144 hr LC50 

(extruded diet)

276 - 

Rainbow trout (10.9 g),Oncorhynchus mykiss

F,M,D Copper 

sulfate

362 144 hr LC50 

(steam pelleted diet)

350 - 

B-21

Appendix B. Other Data on Effects of Copper on Freshwater Organisms
Species Methoda Chemical

Hardness 

(mg/L as 

CaCO3)

Duration Effect

Total

Concentration

(μg/L)b

Dissolved

Concentration

(μg/L)

Rainbow trout (12.3 g),Oncorhynchus mykiss

F,M,D Copper 

sulfate

362 144 hr LC50 

(Low carbohydrate diet)

408 - 

Rainbow trout (11.6 g),Oncorhynchus mykiss

F,M,D Copper 

sulfate

362 144 hr LC50 

(high carbohydrate diet)

246 - 

Rainbow trout (1.7-3.3 g),Oncorhynchus mykiss

F,M,D Copper 

sulfate

374 21 days Incipient lethal level 329 - 

Rainbow trout (1.7-3.3 g),Oncorhynchus mykiss

F,M,D Copper 

sulfate

374 21 days Incipient lethal level 333 - 

Rainbow trout (1.7-3.3 g),Oncorhynchus mykiss

F,M,D Copper 

sulfate

374 21 days Incipient lethal level 311 - 

Rainbow trout (1.7-3.3 g),Oncorhynchus mykiss

F,M,D Copper 

sulfate

374 21 days Incipient lethal level 274 - 

Rainbow trout (1.7-3.3 g),Oncorhynchus mykiss

F,M,D Copper 

sulfate

374 21 days Incipient lethal level 371 - 

Rainbow trout (1.7-3.3 g),Oncorhynchus mykiss

F,M,D Copper 

sulfate

374 21 days Incipient lethal level (acclimated to 30 

ug/L)

266 - 

Rainbow trout (1.7-3.3 g),Oncorhynchus mykiss

F,M,D Copper 

sulfate

374 21 days Incipient lethal level (acclimated to 58 

ug/L)

349 - 

Rainbow trout (1.7-3.3 g),Oncorhynchus mykiss

F,M,D Copper 

sulfate

374 21 days Incipient lethal level (acclimated to 94 

ug/L)

515 - 

Rainbow trout (1.7-3.3 g),Oncorhynchus mykiss

F,M,D Copper 

sulfate

374 21 days Incipient lethal level (acclimated to 131

ug/L)

564 - 

Rainbow trout (1.7-3.3 g),Oncorhynchus mykiss

F,M,D Copper 

sulfate

374 21 days Incipient lethal level (acclimated to 194

ug/L)

708 - 

Rainbow trout (2.9 g),Oncorhynchus mykiss

F,M,D Copper 

chloride

30.5 ca. 2 hr Inhibited avoidance of serine 6.667 - 

Rainbow trout (3.2 g),Oncorhynchus mykiss

F,M,T,D Copper 

sulfate

30 96 hr LC50 - 19.9 

Rainbow trout (1.4 g),Oncorhynchus mykiss

F,M,T,D Copper 

sulfate

101 96 hr LC50 - 176 

Rainbow trout (2.2 g),Oncorhynchus mykiss

F,M,T,D Copper 

sulfate

370 96 hr LC50 - 232 

Rainbow trout (smolt),Oncorhynchus mykiss

F,M,T,D Copper 

sulfate

363 >10 days LC50 97.92 - 

Rainbow trout (parr), Oncorhynchus mykiss

F,M,T,D,I - 31.0 62 days NOEC 

(growth and survival)

90 - 

Atlantic salmon (2-3 yr parr),Salmo salar

S,M,T - 8-10 96 hr LC50 125 - 

Atlantic salmon (6.4-11.7 cm),Salmo salar

F,M,T Copper 

sulfate

20 7 days LC50 48 - 

Atlantic salmon (7.2-10.9 cm),Salmo salar

F,M,T - 14 7 days LC50 32 - 

Brown trout (3-6 day larva),Salmo trutta

S,M,T Copper 

chloride

4 30 days >90% mortality 80 - 

B-22

Appendix B. Other Data on Effects of Copper on Freshwater Organisms
Species Methoda Chemical

Hardness 

(mg/L as 

CaCO3)

Duration Effect

Total

Concentration

(μg/L)b

Dissolved

Concentration

(μg/L)

Brown trout (larva),Salmo trutta

S,M,T Copper 

chloride

4 30 days >90% mortality 20 - 

Brown trout (larva),Salmo trutta

S,M,T Copper 

chloride

22 30 days <10% mortality 80 - 

Brown trout (larva),Salmo trutta

F,M,T Copper 

chloride

25 60 days Inhibited growth 4.6 - 

Brook trout,Salvelinus fontinalis

- - - 24 hr Significant change in cough rate 9 - 

Brook trout (1 g),Salvelinus fontinalis

S,M,T Copper 

chloride

4 80 hr 75% mortality 25.4 - 

Brook trout (8 mo),Salvelinus fontinalis

R,M,T - 20 10 days IC50 

(growth) 

187 - 

Brook trout (15-20 cm),Salvelinus fontinalis

F,M,T Copper 

sulfate

47 21 days Altered Blood Hct, RBC, Hb, Cl, 

PGOT, Osmolarity, protein

38.2 - 

Brook trout (13-20 cm),Salvelinus fontinalis

F,M,T Copper 

sulfate

47 337 days Altered blood PGOT 17.4 - 

Goldfish (3.8-6.3 cm),Carassius auratus

S,U Copper 

sulfate

20 96 hr LC50 36 

Goldfish (10.5 g),Carassius auratus

S,M,T Copper 

sulfate

34.2 - LC50 150 - 

Goldfish (embryo),Carrassius auratus

R,U Copper 

sulfate

195 7 days EC50 

(death or deformity)

5,200 - 

Goldfish,Carassius auratus

R,U Copper 

sulfate

45 24 hr LC50 

(50 C) 

2,700 - 

Goldfish,Carassius auratus

R,U Copper 

sulfate

45 24 hr LC50 

(150 C)

2,900 - 

Goldfish,Carassius auratus

R,U Copper 

sulfate

45 24 hr LC50 

(300 C)

1,510 - 

Common carp (1.8-2.1 cm),Cyprinus carpio

S,U Copper 

sulfate

144-188 96 hr LC50 117.5 

Common carp (5.0-6.0 cm),Cyprinus carpio

S,U Copper 

sulfate

144-188 96 hr LC50 530 

Common carp (embryo),Cyprinus carpio

S,U Copper 

sulfate

360 - EC50

(hatch and deformity)

4,775 - 

Common carp (embryo),Cyprinus carpio

S,U Copper 

acetate

274 96 hr LC50 140 - 

Common carp (larva),Cyprinus carpio

S,U Copper 

acetate

274 96 hr LC50 4 - 

Common carp (fry),Cyprinus carpio

S,U Copper 

acetate

274 96 hr LC50 63 - 

Common carp,Cyprinus carpio

S,M,T Copper 

nitrate

53 - LC50 110 - 

Common carp,Cyprinus carpio

S,M,T Copper 

nitrate

55 - LC50 800 - 

B-23

Appendix B. Other Data on Effects of Copper on Freshwater Organisms
Species Methoda Chemical

Hardness 

(mg/L as 

CaCO3)

Duration Effect

Total

Concentration

(μg/L)b

Dissolved

Concentration

(μg/L)

Common carp (4.7-6.2 cm),Cyprinus carpio

R,U Copper 

sulfate

19 96 hr LC50 63 

Common carp (embryo and larva),Cyprinus carpio

R,U Copper 

sulfate

50 108 hr 77% deformed 10 - 

Common carp (3.5 cm),Cyprinus carpio

R,U Copper 

sulfate

- 96 hr LC50 300 - 

Common carp (6.5 cm),Cyprinus carpio

R,U Copper 

sulfate

- 96 hr LC50 1,000 - 

Common carp (embryo),Cyprinus carpio

R,M,T Copper 

sulfate

50 72 hr Prevented hatching 700 - 

Common carp (1 mo),Cyprinus carpio

R,M,T Copper 

nitrate

84.8 1 wk Raised critical D.O. and altered 

ammonia excretion

14.0 - 

Common carp (22.9 cm),Cyprinus carpio

F,M,T Copper 

chloride

17 48 hr LC50 170 - 

Common carp (embryo and larva),Cyprinus carpio

F,M,T Copper 

chloride

100 168 hr 55% mortality 19 - 

Common carp (embryo and larva),Cyprinus carpio

F,M,T Copper 

chloride

100 168 hr 18% mortality; 50.8 - 

Bonytail (larva),Gila elegans

S, U Copper 

sulfate

199 96 hr LC50 364 

Bonytail (100-110 days),Gila elegans

S, U Copper 

sulfate

199 96 hr LC50 231 

Golden shiner (11-13 cm),Notemigonus crysoleucas

S,U Copper 

sulfate

221 94 hr Decreased serum osmolality 2,500 - 

Golden shiner,Notemigonus crysoleucas

S,U Copper 

sulfate

45 24 hr LC50 

(50 C) 

330 - 

Golden shiner,Notemigonus crysoleucas

S,U Copper 

sulfate

45 24 hr LC50 

(150 C)

230 - 

Golden shiner,Notemigonus crysoleucas

S,U Copper 

sulfate

45 24 hr LC50 

(300 C)

270 - 

Golden shiner,Notemigonus crysoleucas

F,M,T Copper 

chloride

72.2 15 min EC50 

(avoidance)

26 - 

Striped shiner,Notropis chrysocephalus

F,M,T,D Copper 

sulfate

318 96 hr LC50 3,400 - 

Striped shiner (4.7 cm)Notropis chrysocephalus

F,M,T,D Copper 

sulfate

316 96 hr LC50 4,000 - 

Striped shiner (5.0 cm)Notropis chrysocephalus

F,M,T,D Copper 

sulfate

274 96 hr LC50 5,000 - 

Striped shiner,Notropis chrysocephalus

F,M,T,D Copper 

sulfate

314 96 hr LC50 8,400 - 

B-24

Appendix B. Other Data on Effects of Copper on Freshwater Organisms
Species Methoda Chemical

Hardness 

(mg/L as 

CaCO3)

Duration Effect

Total

Concentration

(μg/L)b

Dissolved

Concentration

(μg/L)

Striped shiner,Notropis chrysocephalus

F,M,T,D Copper 

sulfate

303 96 hr LC50 16,000 - 

Bluntnose minnow,Pimephales notatus

S,U Copper 

sulfate

208 48 hr LC50 290 - 

Bluntnose minnow,Pimephales notatus

S,U Copper 

sulfate

132 48 hr LC50 150 - 

Bluntnose minnow,Pimephales notatus

S,U Copper 

sulfate

182 48 hr LC50 200 - 

Bluntnose minnow,Pimephales notatus

S,U Copper 

sulfate

233 48 hr LC50 180 - 

Bluntnose minnow,Pimephales notatus

S,U Copper 

sulfate

282 48 hr LC50 260 - 

Bluntnose minnow,Pimephales notatus

S,U Copper 

sulfate

337 48 hr LC50 260 - 

Bluntnose minnow,Pimephales notatus

S,U Copper 

sulfate

322 48 hr LC50 6,300 - 

Bluntnose minnow,Pimephales notatus

S,U Copper 

sulfate

322 48 hr LC50 11,000 - 

Bluntnose minnow,Pimephales notatus

S,U Copper 

sulfate

322 48 hr LC50 25,000 - 

Bluntnose minnow,Pimephales notatus

S,U Copper 

sulfate

203 48 hr LC50 160 - 

Bluntnose minnow,Pimephales notatus

S,U Copper 

sulfate

203 48 hr LC50 1,100 - 

Bluntnose minnow,Pimephales notatus

S,U Copper 

sulfate

203 48 hr LC50 2,900 - 

Bluntnose minnow,Pimephales notatus

S,M,D Copper 

sulfate

320 48 hr LC50 6,300 - 

Bluntnose minnow,Pimephales notatus

S,M,D Copper 

sulfate

324 48 hr LC50 9,000 - 

Bluntnose minnow,Pimephales notatus

S,M,D Copper 

sulfate

324 48 hr LC50 4,700 - 

Bluntnose minnow,Pimephales notatus

S,M,D Copper 

sulfate

320 48 hr LC50 11,000 - 

Bluntnose minnow,Pimephales notatus

S,M,D Copper 

sulfate

318 48 hr LC50 5,700 - 

Bluntnose minnow,Pimephales notatus

S,M,D Copper 

sulfate

318 48 hr LC50 10,000 - 

Bluntnose minnow,Pimephales notatus

S,M,D Copper 

sulfate

314 48 hr LC50 8,000 - 

Bluntnose minnow,Pimephales notatus

S,M,D Copper 

sulfate

318 48 hr LC50 11,000 - 

Bluntnose minnow,Pimephales notatus

S,M,D Copper 

sulfate

324 48 hr LC50 9,700 - 

B-25

Appendix B. Other Data on Effects of Copper on Freshwater Organisms
Species Methoda Chemical

Hardness 

(mg/L as 

CaCO3)

Duration Effect

Total

Concentration

(μg/L)b

Dissolved

Concentration

(μg/L)

Bluntnose minnow,Pimephales notatus

S,M,D Copper 

sulfate

339 48 hr LC50 7,000 - 

Bluntnose minnow,Pimephales notatus

S,M,D Copper 

sulfate

310 48 hr LC50 12,000 - 

Bluntnose minnow,Pimephales notatus

S,M,D Copper 

sulfate

310 48 hr LC50 21,000 - 

Bluntnose minnow,Pimephales notatus

S,M,D Copper 

sulfate

302 48 hr LC50 19,000 - 

Bluntnose minnow,Pimephales notatus

S,M,D Copper 

sulfate

296 48 hr LC50 8,000 - 

Bluntnose minnow,Pimephales notatus

S,M,D Copper 

sulfate

332 48 hr LC50 11,000 - 

Bluntnose minnow,Pimephales notatus

S,M,D Copper 

sulfate

340 48 hr LC50 6,300 - 

Bluntnose minnow,Pimephales notatus

S,M,D Copper 

sulfate

296 48 hr LC50 1,500 - 

Bluntnose minnow,Pimephales notatus

S,M,D Copper 

sulfate

306 48 hr LC50 750 - 

Bluntnose minnow,Pimephales notatus

S,M,D Copper 

sulfate

308 48 hr LC50 2,500 - 

Bluntnose minnow,Pimephales notatus

S,M,D Copper 

sulfate

304 48 hr LC50 1,600 - 

Bluntnose minnow,Pimephales notatus

S,M,D Copper 

sulfate

315 48 hr LC50 4,000 - 

Bluntnose minnow (3.9 cm),Pimephales notatus

F,M,T,D Copper 

sulfate

314 96 hr LC50 6,800 - 

Bluntnose minnow (5.3 cm),Pimephales notatus

F,M,T,D Copper 

sulfate

303 96 hr LC50 13,000 - 

Fathead minnow (adult),Pimephales promelas

S,U Copper 

sulfate

103-104 96 hr LC50 210

Fathead minnow (adult),Pimephales promelas

S,U Copper 

sulfate

103-104 96 hr LC50 310

Fathead minnow (adult),Pimephales promelas

S,U Copper 

sulfate

103-104 96 hr LC50 120

Fathead minnow (adult),Pimephales promelas

S,U Copper 

sulfate

103-104 96 hr LC50 210 

Fathead minnow (adult),Pimephales promelas

S,U Copper 

sulfate

254-271 96 hr LC50 390 

Fathead minnow,Pimephales promelas

S,U Copper 

sulfate

200 96 hr LC50 430 

Fathead minnow,Pimephales promelas

S,U Copper 

sulfate

31 96 hr LC50 84 

Fathead minnow (3.8-6.3 cm),Pimephales promelas

S,U Copper 

sulfate

20 96 hr LC50 25 

B-26

Appendix B. Other Data on Effects of Copper on Freshwater Organisms
Species Methoda Chemical

Hardness 

(mg/L as 

CaCO3)

Duration Effect

Total

Concentration

(μg/L)b

Dissolved

Concentration

(μg/L)

Fathead minnow (3.8-6.3 cm),Pimephales promelas

S,U Copper 

sulfate

20 96 hr LC50 23 

Fathead minnow (3.8-6.3 cm),Pimephales promelas

S,U Copper 

sulfate

20 96 hr LC50 23 

Fathead minnow (3.8-6.3 cm),Pimephales promelas

S,U Copper 

sulfate

20 96 hr LC50 22 

Fathead minnow (3.8-6.3 cm),Pimephales promelas

S,U Copper 

sulfate

360 96 hr LC50 1760 

Fathead minnow (3.8-6.3 cm),Pimephales promelas

S,U Copper 

sulfate

360 96 hr LC50 1140 

Fathead minnow,Pimephales promelas

S,U Copper 

sulfate

20 96 hr LC50 50 

Fathead minnow,Pimephales promelas

S,U Copper 

sulfate

400 96 hr LC50 1,400 

Fathead minnow (3.2-4.2 cm),Pimephales promelas

S,M Copper 

acetate

44 96 hr LC50 117 - 

Fathead minnow (2.0-6.9 cm),Pimephales promelas

S,M,D Copper 

sulfate

294 96 hr LC50 16,000 - 

Fathead minnow (2.0-6.9 cm),Pimephales promelas

S,M,D Copper 

sulfate

120 96 hr LC50 2,200 - 

Fathead minnow (2.0-6.9 cm),Pimephales promelas

S,M,D Copper 

sulfate

298 96 hr LC50 16,000 - 

Fathead minnow (2.0-6.9 cm),Pimephales promelas

S,M,D Copper 

sulfate

280 96 hr LC50 3,300 - 

Fathead minnow (2.0-6.9 cm),Pimephales promelas

S,M,D Copper 

sulfate

244 96 hr LC50 1,600 - 

Fathead minnow (2.0-6.9 cm),Pimephales promelas

S,M,D Copper 

sulfate

212 96 hr LC50 2,000 - 

Fathead minnow (2.0-6.9 cm),Pimephales promelas

S,M,D Copper 

sulfate

260 96 hr LC50 3,500 - 

Fathead minnow (2.0-6.9 cm),Pimephales promelas

S,M,D Copper 

sulfate

224 96 hr LC50 9,700 - 

Fathead minnow (2.0-6.9 cm),Pimephales promelas

S,M,D Copper 

sulfate

228 96 hr LC50 5,000 - 

Fathead minnow (2.0-6.9 cm),Pimephales promelas

S,M,D Copper 

sulfate

150 96 hr LC50 2,800 - 

Fathead minnow (2.0-6.9 cm),Pimephales promelas

S,M,D Copper 

sulfate

310 96 hr LC50 11,000 - 

Fathead minnow (2.0-6.9 cm),Pimephales promelas

S,M,D Copper 

sulfate

280 96 hr LC50 12,000 - 

Fathead minnow (2.0-6.9 cm),Pimephales promelas

S,M,D Copper 

sulfate

280 96 hr LC50 11,000 - 

B-27

Appendix B. Other Data on Effects of Copper on Freshwater Organisms
Species Methoda Chemical

Hardness 

(mg/L as 

CaCO3)

Duration Effect

Total

Concentration

(μg/L)b

Dissolved

Concentration

(μg/L)

Fathead minnow (2.0-6.9 cm),Pimephales promelas

S,M,D Copper 

sulfate

260 96 hr LC50 22,200 - 

Fathead minnow (2.0-6.9 cm),Pimephales promelas

S,M,D Copper 

sulfate

308 96 hr LC50 4,670 - 

Fathead minnow (2.0-6.9 cm),Pimephales promelas

S,M,D Copper 

sulfate

206 96 hr LC50 920 - 

Fathead minnow (2.0-6.9 cm),Pimephales promelas

S,M,D Copper 

sulfate

262 96 hr LC50 1,190 - 

Fathead minnow (2.0-6.9 cm),Pimephales promelas

S,M,D Copper 

sulfate

322 96 hr LC50 2,830 - 

Fathead minnow (2.0-6.9 cm),Pimephales promelas

S,M,D Copper 

sulfate

210 96 hr LC50 1,450 - 

Fathead minnow (2.0-6.9 cm),Pimephales promelas

S,M,D Copper 

sulfate

260 96 hr LC50 1,580 - 

Fathead minnow (2.0-6.9 cm),Pimephales promelas

S,M,D Copper 

sulfate

252 96 hr LC50 1,000 - 

Fathead minnow (2.0-6.9 cm),Pimephales promelas

S,M,D Copper 

sulfate

312 96 hr LC50 5,330 - 

Fathead minnow (2.0-6.9 cm),Pimephales promelas

S,M,D Copper 

sulfate

276 96 hr LC50 4,160 - 

Fathead minnow (2.0-6.9 cm),Pimephales promelas

S,M,D Copper 

sulfate

252 96 hr LC50 10,550 - 

Fathead minnow (2.0-6.9 cm),Pimephales promelas

S,M,D Copper 

sulfate

298 96 hr LC50 22,200 - 

Fathead minnow (2.0-6.9 cm),Pimephales promelas

S,M,D Copper 

sulfate

282 96 hr LC50 21,800 - 

Fathead minnow (2.0-6.9 cm),Pimephales promelas

S,M,D Copper 

sulfate

284 96 hr LC50 23,600 - 

Fathead minnow (<24 h),Pimephales promelas

S,M,T Copper 

nitrate

290 96 hr LC50 >200 - 

Fathead minnow (<24 h),Pimephales promelas

S,M,T Copper 

sulfate

16.8 96 hr LC50 36.0 - 

Fathead minnow (<24 h),Pimephales promelas

S,M,T Copper 

sulfate

19.0 96 hr LC50 70.3 - 

Fathead minnow (<24 h),Pimephales promelas

S,M,T Copper 

sulfate

19.0 96 hr LC50 85.6 - 

Fathead minnow (<24 h),Pimephales promelas

S,M,T Copper 

sulfate

19.0 96 hr LC50 182.0 - 

Fathead minnow (<24 h; 0.68 mg),Pimephales promelas

S,M,T Copper 

sulfate

17 96 hr LC50 1.99 - 

B-28

Appendix B. Other Data on Effects of Copper on Freshwater Organisms
Species Methoda Chemical

Hardness 

(mg/L as 

CaCO3)

Duration Effect

Total

Concentration

(μg/L)b

Dissolved

Concentration

(μg/L)

Fathead minnow (<24 h; 0.68 mg),Pimephales promelas

S,M,T Copper 

sulfate

20.5 96 hr LC50 4.86 - 

Fathead minnow (<24 h; 0.68 mg),Pimephales promelas

S,M,T Copper 

sulfate

16.5 96 hr LC50 11.1 - 

Fathead minnow (<24 h; 0.68 mg),Pimephales promelas

S,M,T Copper 

sulfate

17.5 96 hr LC50 9.87 - 

Fathead minnow (<24 h; 0.68 mg),Pimephales promelas

S,M,T Copper 

sulfate

17 96 hr LC50 15.7 - 

Fathead minnow (60-90 days),Pimephales promelas

S,M,T - 110 48 hr LC50 284 - 

Fathead minnow (3 wk),Pimephales promelas

S,M,T Copper 

sulfate

101 48 hr Short-term intolerance of hypoxia (2 

mg D.O./L)

186 - 

Fathead minnow (2-4 day),Pimephales promelas

S,M,T Copper 

sulfate

6-10 - LC50 12.5 - 

Fathead minnow (<24 hrs),Pimephales promelas

S,M,T Copper 

sulfate

9.9 96 hr LC50 10.7 - 

Fathead minnow (<24 hrs),Pimephales promelas

S,M,T Copper 

sulfate

7.1 96 hr LC50 6.3 - 

Fathead minnow (<24 hrs),Pimephales promelas

S,M,T Copper 

sulfate

8.3 96 hr LC50 12.2 - 

Fathead minnow (<24 hrs),Pimephales promelas

S,M,T Copper 

sulfate

8.9 96 hr LC50 9.5 - 

Fathead minnow (<24 hrs),Pimephales promelas

S,M,T Copper 

sulfate

16.8 96 hr LC50 26.8 - 

Fathead minnow (<24 hrs),Pimephales promelas

S,M,T Copper 

sulfate

12.2 96 hr LC50 21.2 - 

Fathead minnow (<24 hrs),Pimephales promelas

S,M,T Copper 

sulfate

9.4 96 hr LC50 19.8 - 

Fathead minnow (<24 hrs),Pimephales promelas

S,M,T Copper 

sulfate

11.4 96 hr LC50 31.9 - 

Fathead minnow (<24 hrs),Pimephales promelas

S,M,T Copper 

sulfate

10.9 96 hr LC50 26.1 - 

Fathead minnow (<24 hrs),Pimephales promelas

S,M,T Copper 

sulfate

12.4 96 hr LC50 26.0 - 

Fathead minnow (<24 hrs),Pimephales promelas

S,M,T Copper 

sulfate

17.4 96 hr LC50 169.5 - 

Fathead minnow (<24 hrs),Pimephales promelas

S,M,T,D Copper 

sulfate

46 96 hr LC50 17.15 14.87 

Fathead minnow (<24 hrs),Pimephales promelas

S,M,T,D Copper 

sulfate

46 96 hr LC50 21.59 18.72 

B-29

Appendix B. Other Data on Effects of Copper on Freshwater Organisms
Species Methoda Chemical

Hardness 

(mg/L as 

CaCO3)

Duration Effect

Total

Concentration

(μg/L)b

Dissolved

Concentration

(μg/L)

Fathead minnow (<24 hrs),Pimephales promelas

S,M,T,D Copper 

sulfate

47 96 hr LC50 123.19 106.8 

Fathead minnow (<24 hrs),Pimephales promelas

S,M,T,D Copper 

sulfate

45 96 hr LC50 42.56 36.89 

Fathead minnow (<24 hrs),Pimephales promelas

S,M,T,D Copper 

sulfate

46 96 hr LC50 83.19 72.13 

Fathead minnow,Pimephales promelas

S,M,T,D Copper 

sulfate

100 96 hr LC50 (fish from metal-contaminated 

pond)

360 - 

Fathead minnow,Pimephales promelas

S,M,T,D Copper 

sulfate

250 96 hr LC50 (fish from metal-contaminated 

pond)

410 - 

Fathead minnow (<24 hr),Pimephales promelas

R,U - 45 7 days LC50 70 - 

Fathead minnow (<24 hr),Pimephales promelas

R,U - 45 7 days LOEC

(growth)

26 - 

Fathead minnow (<24 hr),Pimephales promelas

R,U Copper 

sulfate

345 4 days RNA threshhold effect 130 - 

Fathead minnow (embryo),Pimephales promelas

R,U Copper 

sulfate

106 5 days LC50 480 - 

Fathead minnow (embryo),Pimephales promelas

R,U Copper 

sulfate

106 5 days LC50 440 - 

Fathead minnow (embryo),Pimephales promelas

R,U Copper 

sulfate

106 5 days EC50

(malformation)

270 - 

Fathead minnow (embryo),Pimephales promelas

R,U Copper 

sulfate

106 5 days EC50

(malformation)

260 - 

Fathead minnow (embryo),Pimephales promelas

R,U Copper 

sulfate

106 7 days LC50 310 - 

Fathead minnow (embryo),Pimephales promelas

R,U Copper 

sulfate

106 7 days LC50 330 - 

Fathead minnow (embryo),Pimephales promelas

R,U Copper 

sulfate

106 7 days EC50

(malformation)

190 - 

Fathead minnow (embryo),Pimephales promelas

R,U Copper 

sulfate

106 7 days EC50

(malformation)

170 - 

Fathead minnow (embryo),Pimephales promelas

R,U Copper 

sulfate

106 7 days LOEC

(length)

160 - 

Fathead minnow (embryo),Pimephales promelas

R,U Copper 

sulfate

106 7 days LOEC

(length)

180 - 

Fathead minnow (larva),Pimephales promelas

R,M,T Copper 

sulfate

180 7 days LOEC 

(growth)

25 - 

Fathead minnow (larva),Pimephales promelas

R,M,T Copper 

sulfate

218 7 days LOEC 

(growth)

38 - 

Fathead minnow (larva),Pimephales promelas

R,M,T Copper 

sulfate

218 7 days LOEC 

(growth)

38 - 

Fathead minnow (3-7 days),Pimephales promelas

R,M,T Copper 

sulfate

74 48 hr LC50 225 - 

B-30

Appendix B. Other Data on Effects of Copper on Freshwater Organisms
Species Methoda Chemical

Hardness 

(mg/L as 

CaCO3)

Duration Effect

Total

Concentration

(μg/L)b

Dissolved

Concentration

(μg/L)

Fathead minnow (larva),Pimephales promelas

R,M,T,D Copper 

sulfate

80 48 hr LC50 35.9 - 

Fathead minnow (larva),Pimephales promelas

R,M,T,D Copper 

sulfate

80 48 hr LC50 28.9 - 

Fathead minnow (larva),Pimephales promelas

R,M,T,D Copper 

sulfate

80 48 hr LC50 20.7 - 

Fathead minnow (larva),Pimephales promelas

R,M,T,D Copper 

sulfate

80 48 hr LC50 80.8 - 

Fathead minnow (3-7 days),Pimephales promelas

R,M,T,D Copper 

sulfate

80 48 hr LC50 297.1 - 

Fathead minnow (3-7 days),Pimephales promelas

R,M,T,D Copper 

sulfate

72 48 hr LC50 145.8 - 

Fathead minnow (32-38 mm),Pimephales promelas

F,M,T Copper 

sulfate

244 9 mo LOEC

(93% lower fecundity)

120 - 

Fathead minnow (larva),Pimephales promelas

F,M,T Copper 

sulfate

202 - LC50 250 - 

Fathead minnow (embryo),Pimephales promelas

F,M,T Copper 

sulfate

202 34 days Reduced growth;

increased abnormality

61 - 

Fathead minnow (embryo),Pimephales promelas

F,M,T Copper 

sulfate

202 34 days LC50 123 - 

Fathead minnow (24-96 hr),Pimephales promelas

F,M,T Copper 

sulfate

10.7 21 days Incipient lethal level 6.2 - 

Fathead minnow (24-96 hr),Pimephales promelas

F,M,T Copper 

sulfate

10.7 21 days Growth (length) reduced by 8% 5.3 - 

Fathead minnow (24-96 hr),Pimephales promelas

F,M,T Copper 

sulfate

9.3 21 days Incipient lethal level 17.2 - 

Fathead minnow (24-96 hr),Pimephales promelas

F,M,T Copper 

sulfate

9.3 21 days Growth (length) reduced by 17% 16.2 - 

Fathead minnow (<24 hrs),Pimephales promelas

F,M,T Copper 

sulfate

46 96 hr LC50 305 - 

Fathead minnow (<24 hrs),Pimephales promelas

F,M,T Copper 

sulfate

46 96 hr LC50 298.6 - 

Fathead minnow,Pimephales promelas

F,M,T - 30 96 hr LC50 

(TOC=12 mg/L)

436 - 

Fathead minnow,Pimephales promelas

F,M,T - 37 96 hr LC50 

(TOC=13 mg/L)

516 - 

Fathead minnow,Pimephales promelas

F,M,T - 87 96 hr LC50 

(TOC=36 mg/L)

1,586 - 

Fathead minnow,Pimephales promelas

F,M,T - 73 96 hr LC50 

(TOC=28 mg/L)

1,129 - 

Fathead minnow,Pimephales promelas

F,M,T - 84 96 hr LC50 

(TOC=15 mg/L)

550 - 

Fathead minnow,Pimephales promelas

F,M,T - 66 96 hr LC50 

(TOC=34 mg/L)

1,001 - 

B-31

Appendix B. Other Data on Effects of Copper on Freshwater Organisms
Species Methoda Chemical

Hardness 

(mg/L as 

CaCO3)

Duration Effect

Total

Concentration

(μg/L)b

Dissolved

Concentration

(μg/L)

Fathead minnow,Pimephales promelas

F,M,T - 117 96 hr LC50 

(TOC=30 mg/L)

2,050 - 

Fathead minnow,Pimephales promelas

F,M,T - 121 96 hr LC50 

(TOC=30 mg/L)

2,336 - 

Fathead minnow,Pimephales promelas

F,M,T Copper 

sulfate

117 96 hr LC50 2,050 - 

Fathead minnow,Pimephales promelas

F,M,T Copper 

sulfate

121 96 hr LC50 2,336 - 

Fathead minnow (4.4 cm),Pimephales promelas

F,M,T,D Copper 

sulfate

314 96 hr LC50 11,000 - 

Fathead minnow (4.2 cm),Pimephales promelas

F,M,T,D Copper 

sulfate

303 96 hr LC50 15,000 - 

Fathead minnow (<24 hrs),Pimephales promelas

F,M,T,D Copper 

sulfate

45 96 hr LC50 158.8 138.1 

Fathead minnow (<24 hrs),Pimephales promelas

F,M,T,D Copper 

sulfate

45 96 hr LC50 80.01 72.01 

Fathead minnow (<24 hrs),Pimephales promelas

F,M,T,D Copper 

sulfate

46 96 hr LC50 20.96 18.23 

Fathead minnow (<24 hrs),Pimephales promelas

F,M,T,D Copper 

sulfate

44 96 hr LC50 50.8 39.12 

Fathead minnow (<24 hrs),Pimephales promelas

F,M,T,D Copper 

sulfate

45 96 hr LC50 65.41 45.78 

Colorado squawfish (larva),Ptychocheilus lucius

S,U Copper 

sulfate

199 96 hr LC50 363 

Colorado squawfish (155-186 days),Ptychocheilus lucius

S,U Copper 

sulfate

199 96 hr LC50 663 

Colorado squawfish (32-40 days posthatch),Ptychocheilus lucius

S,U Copper 

sulfate

144 96 hr LC50 293 

Colorado squawfish (32-40 days posthatch),Ptychocheilus lucius

S,U Copper 

sulfate

144 96 hr LC50 320 

Creek chub,Semotilus atromaculatus

F,M,T Copper 

sulfate

316 96 hr LC50 11,500 - 

Creek chub,Semotilus atromaculatus

F,M,T Copper 

sulfate

274 96 hr LC50 1,100 - 

Razorback sucker (larva),Xyrauchen texanus

S,U Copper 

sulfate

199 96 hr LC50 404 

Razorback sucker (102-116 days),Xyrauchen texanus

S,U Copper 

sulfate

199 96 hr LC50 331 

Razorback sucker (13-23 days posthatch),Xyrauchen texanus

S,U Copper 

sulfate

144 96 hr LC50 231 

B-32

Appendix B. Other Data on Effects of Copper on Freshwater Organisms
Species Methoda Chemical

Hardness 

(mg/L as 

CaCO3)

Duration Effect

Total

Concentration

(μg/L)b

Dissolved

Concentration

(μg/L)

Razorback sucker (13-23 days posthatch),Xyrauchen texanus

S,U Copper 

sulfate

144 96 hr LC50 314 

Brown bullhead,Ictallurus nebulosus

F,M,T Copper 

sulfate

303 96 hr LC50 12,000 - 

Brown bullhead (5.2 cm),Ictalurus nebulosus

F,M,T Copper 

sulfate

314 96 hr LC50 5,200 - 

Channel catfish (13-14 cm),Ictalurus punctatus

S,U Copper 

sulfate

221 94 hr Decreased serum osmolality 2,500 - 

Channel catfish,Ictalurus punctatus

S,U Copper 

sulfate

45 24 hr LC50 

(50 C)

3,700 - 

Channel catfish,Ictalurus punctatus

S,U Copper 

sulfate

45 24 hr LC50 

(150 C)

2,600 - 

Channel catfish,Ictalurus punctatus

S,U Copper 

sulfate

45 24 hr LC50 

(300 C)

3,100 - 

Channel catfish,Ictalurus punctatus

S,U Copper 

sulfate

100 10 days EC50 

(death and deformity)

6,620 - 

Channel catfish (fingerlings),Ictalurus punctatus

S,U Copper 

sulfate

16 96 hr LC50 54 

Channel catfish (fingerlings),Ictalurus punctatus

S,U Copper 

sulfate

16 96 hr LC50 55 

Channel catfish (fingerlings),Ictalurus punctatus

S,U Copper 

sulfate

83 96 hr LC50 762 

Channel catfish (fingerlings),Ictalurus punctatus

S,U Copper 

sulfate

83 96 hr LC50 700 

Channel catfish (fingerlings),Ictalurus punctatus

S,U Copper 

sulfate

161 96 hr LC50 768 

Channel catfish (fingerlings),Ictalurus punctatus

S,U Copper 

sulfate

161 96 hr LC50 1139 

Channel catfish (fingerlings),Ictalurus punctatus

S,U Copper 

sulfate

287 96 hr LC50 1041 

Channel catfish (fingerlings),Ictalurus punctatus

S,U Copper 

sulfate

287 96 hr LC50 925 

Channel catfish (400-600 g),Ictalurus punctatus

F,M,T Copper 

sulfate

- 10 wk Significant mortality 354 - 

Channel catfish (4.1 gm),Ictalurus punctatus

F,M,T,D Copper 

sulfate

319 14 days LC50 1,229 - 

Channel catfish (5.7 gm),Ictalurus punctatus

F,M,T,D Copper 

sulfate

315 14 days LC50 1,073 - 

Banded killifish,Fundulus diaphanus

S,M,T Copper 

nitrate

53 - 860 - 

Banded killifish,Fundulus diaphanus

S,M,T Copper 

nitrate

55 - 840 - 

B-33

Appendix B. Other Data on Effects of Copper on Freshwater Organisms
Species Methoda Chemical

Hardness 

(mg/L as 

CaCO3)

Duration Effect

Total

Concentration

(μg/L)b

Dissolved

Concentration

(μg/L)

Flagfish (0.1-0.3 g),Jordanella floridae

F,M,T,D Copper 

sulfate

363 10 days LC50 - 680 

Flagfish (0.1-0.3 g),Jordanella floridae

F,M,T,D Copper 

sulfate

363 96 hr LC50 - 1,270 

Mosquitofish (3.8-5.1 cm female),Gambusia affinis

S,U Copper 

nitrate

27-41 96 hr LC50 93 

Mosquitofish (3.8-5.1 cm female),Gambusia affinis

S,U Copper 

sulfate

27-41 96 hr LC50 200 

Mosquitofish (2.5 cm male),Gambusia affinis

S,U - 50 96 hr LC50 3,500 

Mosquitofish (2.5 cm male),Gambusia affinis

S,U - 150 96 hr LC50 5,000 

Mosquitofish (2.5 cm male),Gambusia affinis

S,U - 300 96 hr LC50 6,000 

Mosquitofish (3.5 cm female),Gambusia affinis

S,U - 50 96 hr LC50 2,500 

Mosquitofish (3.5 cm female),Gambusia affinis

S,U - 150 96 hr LC50 2,900 

Mosquitofish (3.5 cm female),Gambusia affinis

S,U - 300 96 hr LC50 5,000 

Mosquitofish (0.8 cm fry),Gambusia affinis

S,U - 50 96 hr LC50 900 

Mosquitofish (0.8 cm fry),Gambusia affinis

S,U - 150 96 hr LC50 1,400 

Mosquitofish (0.8 cm fry),Gambusia affinis

S,U - 300 96 hr LC50 2,000 

Mosquito fish,Gambusia affinis

S,U Copper 

sulfate

- 96 hr LC50 

(high turbidity)

75,000 - 

Mosquito fish,Gambusia affinis

R,M Copper 

sulfate

45 48 hr LC50 180 - 

Guppy (1.5 cm),Poecilia reticulata

S,U Copper 

sulfate

230 96 hr LC50 1,230 

Guppy (1.62 cm),Poecilia reticulata

S,U Copper 

sulfate

240 96 hr LC50 764 

Guppy (1.9-2.5 cm),Poecilia reticulata

S,U Copper 

sulfate

20 96 hr LC50 36 

Guppy (1.5 cm),Poecilia reticulata

R,U Copper 

sulfate

260 96 hr LC50 2,500 

Guppy (0.8-1.0 cm),Poecilia reticulata

R,U Copper 

sulfate

144-188 96 hr LC50 160 

Guppy (1.2-2.3 cm; female),Poecilia reticulata

R,U Copper 

sulfate

144-188 96 hr LC50 275 

B-34

Appendix B. Other Data on Effects of Copper on Freshwater Organisms
Species Methoda Chemical

Hardness 

(mg/L as 

CaCO3)

Duration Effect

Total

Concentration

(μg/L)b

Dissolved

Concentration

(μg/L)

Guppy (2.3-2.8 cm; male),Poecilia reticulata

R,U Copper 

sulfate

144-188 96 hr LC50 210 

Guppy (340 mg; female),Poecilia reticulata

R,U Copper 

sulfate

144-188 96 hr LC50 480 

Guppy (1.5 cm),Poecilia reticulata

R,U Copper 

sulfate

260 48 hr LC50 2,500 - 

Guppy (1.5 cm),Poecilia reticulata

R, U Copper 

sulfate

181 96 hr LC50 986 - 

Guppy (1 mo),Poecilia reticulata

F,U Copper 

sulfate

76 24 hr LC50 1,370 - 

Guppy (1 mo),Poecilia reticulata

F,U Copper 

sulfate

76 24 hr LC50 930 - 

Guppy (1 mo),Poecilia reticulata

F,U Copper 

sulfate

76 24 hr LC50 1,130 - 

White perch,Morone americana

S,M,T Copper 

nitrate

53 - LC50 6,200 - 

White perch,Morone americana

S,M,T Copper 

nitrate

55 - LC50 6,400 - 

Striped bass (larva),Morone saxitilis

S,U Copper 

chloride

34.6 96 hr LC50 50 

Striped bass (larva),Morone saxitilis

S,U Copper 

sulfate

34.6 96 hr LC50 100 

Striped bass (3.5-5.1 cm),Morone saxitilis

S,U Copper 

chloride

34.6 96 hr LC50 50 

Striped bass (3.1-5.1 cm),Morone saxitilis

S,U Copper 

sulfate

34.6 96 hr LC50 150 

Striped bass (35-80 day),Morone saxitilis

S,U Copper 

sulfate

285 96 hr LC50 270 

Striped bass (6 cm),Morone saxitilis

S,U Copper 

sulfate

35 96 hr LC50 620 

Striped bass,Morone saxitilis

S,M,T Copper 

nitrate

53 96 hr LC50 4,300 - 

Striped bass,Morone saxitilis

S,M,T Copper 

nitrate

55 96 hr LC50 2,700 - 

Rock bass,Ambloplites rupestris

F,M,T - 24 96 hr LC50 

(high TOC)

1,432 - 

Pumpkinseed (1.2 g),Lepomis gibbosus

S,M,T Copper 

nitrate

53 - LC50 2,400 - 

Pumpkinseed (1.2 g),Lepomis gibbosus

S,M,T Copper 

nitrate

55 - LC50 2,700 - 

Pumpkinseed,Lepomis gibbosus

S,M,T Copper 

nitrate

53 96 hr LC50 2,400 - 

Pumpkinseed,Lepomis gibbosus

S,M,T Copper 

nitrate

55 96 hr LC50 2,700 - 

B-35

Appendix B. Other Data on Effects of Copper on Freshwater Organisms
Species Methoda Chemical

Hardness 

(mg/L as 

CaCO3)

Duration Effect

Total

Concentration

(μg/L)b

Dissolved

Concentration

(μg/L)

Bluegill,Lepomis macrochirus

S,U Copper 

chloride

43 96 hr LC50 770 

Bluegill,Lepomis macrochirus

S,U Copper 

sulfate

43 96 hr LC50 1,250 

Bluegill,Lepomis macrochirus

S,U Copper 

sulfate

45 24 hr LC50 

(50 C)

2,590 - 

Bluegill,Lepomis macrochirus

S,U Copper 

sulfate

45 24 hr LC50 

(150 C)

2,500 - 

Bluegill,Lepomis macrochirus

S,U Copper 

sulfate

45 24 hr LC50 

(300 C)

3,820 - 

Bluegill (3-4 cm),Lepomis macrochirus

S,U - 119 8 days 33% reduction in locomotor activity 40 - 

Bluegill (4.2 cm),Lepomis macrochirus

S,U Copper 

sulfate

52 96 hr LC50 254 

Bluegill (4.2 cm),Lepomis macrochirus

S,U Copper 

sulfate

209 96 hr LC50 437 

Bluegill (4.2 cm),Lepomis macrochirus

S,U Copper 

sulfate

365 96 hr LC50 648 

Bluegill (5-15 g),Lepomis macrochirus

S,U Copper 

sulfate

35 2-6 days 8% increase in oxygen consumption 

rates

300 - 

Bluegill (3.8-6.3 cm),Lepomis macrochirus

S,U Copper 

sulfate

20 96 hr LC50 660 

Bluegill (3.8-6.3 cm),Lepomis macrochirus

S,U Copper 

sulfate

360 96 hr LC50 10,200 

Bluegill,Lepomis macrochirus

S,U Copper 

sulfate

20 96 hr LC50 200 

Bluegill,Lepomis macrochirus

S,U Copper 

sulfate

400 96 hr LC50 10,000 

Bluegill (5-11 cm),Lepomis macrochirus

S,U Copper 

sulfate

46 48 hr LC50 3,000 - 

Bluegill (5-11 cm),Lepomis macrochirus

S,U Copper 

sulfate

101.2 48 hr LC50 7,000 - 

Bluegill (0.51g),Lepomis macrochirus

S,M,T - 110 48 hr LC50 4,300 - 

B-36

Appendix B. Other Data on Effects of Copper on Freshwater Organisms
Species Methoda Chemical

Hardness 

(mg/L as 

CaCO3)

Duration Effect

Total

Concentration

(μg/L)b

Dissolved

Concentration

(μg/L)

Bluegill (5-9 cm),Lepomis macrochirus

S,M,T Copper 

chloride

45-47 - LC50 710 - 

Bluegill (5-9 cm),Lepomis macrochirus

S,M,T Copper 

sulfate

45-47 - LC50 770 - 

Bluegill (5-15 g),Lepomis macrochirus

F,M Copper 

sulfate

35 - LC50 2400 - 

Bluegill (3.5-6.0 cm),Lepomis macrochirus

F,M,T Copper 

sulfate

112.4 80 min Avoidance threshold 8,480 - 

Bluegill (3.2-6.7 cm),Lepomis macrochirus

F,M,T Copper 

chloride

21.2-59.2 96 hr LC50 1,100 - 

Bluegill (3.2-6.7 cm),Lepomis macrochirus

F,M,T Copper 

chloride

21.2-59.2 96 hr LC50 900 - 

Bluegill (35.6-62.3 g),Lepomis macrochirus

F,M,T Copper 

sulfate

273.3 24-96 hr Various behavioral changes 34 - 

Bluegill,Lepomis macrochirus

F,M,T Copper 

chloride

157 24-96 hr 27% reduction in food consumption 31 - 

Bluegill,Lepomis macrochirus

F,M,T,D Copper 

sulfate

316 96 hr LC50 

(high BOD)

16,000 - 

Bluegill,Lepomis macrochirus

F,M,T,D Copper 

sulfate

318 96 hr LC50 (high BOD) 17,000 - 

Bluegill (0.14-0.93 g),Lepomis macrochirus

F,M,T,D Copper 

sulfate

246 14 days LC50 - 2,500 

Bluegill (1.15-2.42 g),Lepomis macrochirus

F,M,T,D Copper 

sulfate

237 14 days LC50 - 3,700 

Bluegill (48.3 g),Lepomis macrochirus

F,M,T,D Copper 

sulfate

40 96 hr Biochemical changes 2,000 - 

Largemouth bass (embryo),Micropterus salmoides

R,U Copper 

sulfate

100 8 days EC50 

(death and deformity)

6,560 - 

Largemouth bass,Micropterus salmoides

F,U - - 24 hr Affected opercular rhythm 48 - 

Rainbow darter,Etheostoma caeruleum

F,M,T,D Copper 

sulfate

318 96 hr LC50 

(high BOD)

4,500 - 

Rainbow darter,Etheostoma caeruleum

F,M,T,D Copper 

sulfate

316 96 hr LC50 

(high BOD)

8,000 - 

Rainbow darter,Etheostoma caeruleum

F,M,T,D Copper 

sulfate

274 96 hr LC50 

(high BOD)

2,800 - 

Rainbow darter (4.6 cm),Etheostoma caeruleum

F,M,T,D Copper 

sulfate

314 96 hr LC50 (high BOD) 4,800 - 

Rainbow darter (4.6 cm),Etheostoma caeruleum

F,M,T,D Copper 

sulfate

303 96 hr LC50 (high BOD) 5,300 - 

Fantail,Etheostoma flabellare

S,M,T Copper 

sulfate

170 96 hr Lowered critical thermal maximum 43 - 

B-37

Appendix B. Other Data on Effects of Copper on Freshwater Organisms
Appendix C. Estimation of Water Chemistry Parameters for 

Acute Copper Toxicity Tests
FINAL REPORT

ESTIMATION OF WATER CHEMISTRY PARAMETERS FOR 

ACUTE COPPER TOXICITY TESTS

For:

U.S. Environmental Protection Agency

Health and Ecological Criteria Division

Office of Science and Technology, Office of Water

1200 Pennsylvania Avenue, NW 

Washington, DC 20460

Great Lakes Environmental Center Program Manager

G.M. DeGraeve

Great Lakes Environmental Center, Inc.

739 Hastings Street, Traverse City, Michigan 49686

Phone: (231) 941-2230
C-1

CONTENTS

Foreword .................................................................... C -3

1.0 Data Acquisition ............................................................ C-4

2.0 Technical Issues and Corresponding Recommendations .................................. C-4

2.1 Estimating Ion Concentrations ............................................. C-4

2.2 pH Adjustment with HCl . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . C-7

2.3 Estimation of DOC ..................................................... C-7

2.4 DOC in Lake Superior Water . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . C-8

2.5 Applying Water Chemistry Data to Lake Superior Water . . . . . . . . . . . . . . . . . . . . . . . . . . . . C-8

2.6 Predicting Ionic Composition of WFTS Well Water . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . C-9

2.7 Data for Measurement of Blacksburg/New River Water . . . . . . . . . . . . . . . . . . . . . . . . . . . . C-11

2.8 Cu Concentrations and Alkalinity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . C-12

2.9 Calculation of DOC and Humic Acid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . C-14

2.10 Alkalinity of Lake Superior Water . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . C-15

2.11 Availability of LC50s .................................................. C-15

2.12 Cl and Na Concentrations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . C-15

2.13 Calculating DOC in Dilution Water . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . C-16

2.14 Ionic Composition of Chehalis River Water . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . C-16

2.15 Chemistry of Water in Howarth and Sprague (1978) .............................. C-17

2.16 Default Values of Analyte Concentrations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . C-18

2.17 Organic Carbon Content of Samples ......................................... C-19

2.18 Additional Water Chemistry Data Needed . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . C-19

2.19 Estimating Data for Waters ............................................... C-19

References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . C-32

Appendix C-1, Calculations for Ionic Composition of Standard Laboratory Reconstituted 

 Water . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . C-35

Appendix C-2, Dissolved, Particulate, and Estimated Total Organic Carbon for Streams and 

 Lakes by State . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . C-36

Appendix C-3, Mean TOC and DOC in Lake Superior Dilution Water . . . . . . . . . . . . . . . . . . . . . . . . . . C-38

Appendix C-4, Measured Hardness and Major Ion and Cation Concentrations in WFTS 

 Well Water from April 1972 to April 1978 ................................ C-39

Appendix C-5, Analytical Results of New and Clinch Rivers and 

 Sinking Creek, VA, Water Samples . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . C-41

Appendix C-6, Water Composition of St. Louis River, MN, from USGS NASQAN and Select 

 Relationships to Water Hardness . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . C-42

Appendix C-7, Supplementary Data for Bennett et al. (1995) ................................. C -48

Appendix C-8, Supplementary Data for Richards and Beitinger (1995) .......................... C -51

Appendix C-9, Water Quality Data for the American River, CA for 

 July 1978 Through December 1980 ...................................... C-52

Appendix C-10, STORET Data for Minnesota Lakes and Rivers .............................. C -53
C-2

TABLES

Table 1. Standard Reconstituted Water Composition and Target Water Quality Characteristics ......... C-5

Table 2. Calculated Ion Concentrations Based on the Standard Salts Added ...................... C -6

Table 3. Adjusted Ion Concentrations for a Standard Reconstituted Water Mix Based on Reported

Hardness ............................................................. C -6

Table 4. Recommended Spreadsheet Addition for Lake Superior Dilution Water . . . . . . . . . . . . . . . . . . . C-9

Table 5. Predicted Ion Concentrations in WFTS Well Water Based on Measured Hardness . . . . . . . . . . . C-10

Table 6. Comparison of Values for Untreated (Natural) and Treated (Dechlorinated City of 

Blacksburg, VA) New River Water . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . C-13

Table 7. Estimated Alkalinity in Natural Surface Water Based on pH .......................... C -14

Table 8. Estimates of Dissolved Organic Carbon and Percent Humic Acid for the Winner (1985)

Toxicity Tests ........................................................ C -14

Table 9. Example Calculations to Estimate Water Chemistry of Tests Conducted at 100 mg/L as 

CaCo3 by Howarth and Sprague (1978) Using a Mixture of University of Guelph Well 

Water and De-ionized Water . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . C-18

FIGURES

Figure 1. Relationship Between Ca and Hardness in WFTS Well Water ........................ C -24

Figure 2. Relationship Between Mg and Hardness in WFTS Well Water . . . . . . . . . . . . . . . . . . . . . . . C-25

Figure 3. Relationship Between Na and Hardness in WFTS Well Water . . . . . . . . . . . . . . . . . . . . . . . . C-26

Figure 4. Relationship Between K and Hardness in WFTS Well Water ........................ C -27

Figure 5. Relationship Between Cl and Hardness in WFTS Well Water . . . . . . . . . . . . . . . . . . . . . . . . C-28

Figure 6. Relationship Between SO4 and Hardness in WFTS Well Water . . . . . . . . . . . . . . . . . . . . . . . C-29

Figure 7. Slopes of the Regression Equations Derived for Na Concentration in St. Louis River, MN,

Water Versus Water Hardness from 1973 to 1993 ................................. C -30

Figure 8. Intercepts of the Regression Equations Derived for Na Concentration in St. Louis River, 

MN, Water Versus Water Hardness from 1973 to 1993 ............................. C-31
C-3

FOREWORD

This report was developed by the Great Lakes Environmental Center. Some minor revisions

were made by the U.S. Environmental Protection Agency (EPA). These revisions were primarily

editorial. Additional editorial and formatting revisions were made by the CDM Group, Inc.

The purpose of this report is to provide input water chemistry information for a Biotic Ligand

Model (BLM) analysis of the acute copper toxicity data in Table 1a of the U.S. Environmental

Protection Agency’s (EPA) draft 2003 Update of Ambient Water Quality Criteria for Copper. EPA

will use these BLM data to derive adjusted aquatic life criteria for copper. Many of the reported Table

1a acute copper toxicity data lack sufficient information on the chemistry of the dilution water to

generate BLM-derived critical accumulation values. This compendium contains data from the

primary authors of these articles. It also contains recommendations for the use of these data,

additional supporting documentation and/or computations, and recommendations for estimating

missing parameters.
C-4

Estimation of Water Chemistry Parameters for Acute Copper Toxicity Tests

To prepare for the possibility of incorporating the Biotic Ligand Model (BLM) (Di Toro et al.

2001) into an updated copper aquatic life criteria document, the U.S. Environmental Protection

Agency (EPA) sought to generate a data table summarizing the acute toxicity of copper to

freshwater organisms that included the following parameters: alkalinity, dissolved organic carbon

(DOC), pH, and the major anions (Cl and SO4) and cations (Ca, Mg, Na, K) of the test water.

Published literature was reviewed and appropriate information tabulated, but measurements for many

of the aforementioned parameters were not reported. To resolve the overwhelming number of

missing test water chemistry values in the database, certain authors were contacted for additional

information and to obtain additional measurements in waters where critical information was either

not measured or not reported. EPA also attempted to determine appropriate methods for estimating

test water chemistry in the absence of reported values. The information received from the authors

and recommended procedures for estimating missing parameters are the subject of this report.

1.0 Data Acquisition

The authors of several studies were contacted for additional information on the chemistry of

the water or methods used in their studies. If the primary or corresponding authors could not be

contacted, an attempt was made to contact secondary authors or personnel from the laboratories

where the studies had been conducted. In a few instances, this initial effort failed to produce the

desired information, and censored databases (U.S. Geological Survey’s [USGS] National Stream

Quality Accounting Network [NASQAN] and EPA’s STOrage and RETrieval [STORET] data

warehouse) were consulted to obtain the missing data. As a last resort, other available sources of

water compositional data (e.g., city drinking water treatment officials) were contacted.

The acquired data were scrutinized for representativeness and usefulness in estimating surrogate

values to complete the water quality information in the original studies. Summary tables and figures

generated from these data are included in the following pages, which serve as the basis for the

addition of values in the spreadsheets. Information used for the tabular and graphical summaries of

these data is included in separate appendices. 

2.0 Technical Issues and Corresponding Recommendations

2.1 Estimating Ion Concentrations

Develop a methodology for estimating Ca, Mg, Na, K, Cl, and SO4 concentrations in

laboratory-reconstituted waters.

Recommendation: The best approach for estimating ion concentrations in standard

laboratory-reconstituted water involves scaling default ion concentrations based on measured

hardness. The default ion concentrations can be computed from the concentrations of the salts

added. The use of calculated ion concentrations as input for the BLM applies only to reconstituted

water prepared following the standard recipes reported in guidance documents for conducting acute

bioassays with aquatic organisms (ASTM 2000; U.S. EPA 1993) (see Table 1). If similar salts are

added in different amounts, then the ion concentrations must be calculated using the recipe reported
C-5

in the article. Otherwise, specific ion ratios, and more importantly ion concentrations, cannot be

calculated.

Table 1. Standard Reconstituted Water Composition and Target Water Quality

Characteristics

Water Type

Reagent Added (mg/L) Final Water Quality

NaHCO3 CaSO4C2H2O MgSO4 KCl pHa Hardnessa Alkalinityb

Very Soft 12.0 7.5 7.5 0.5 6.4-6.8 10-13 10-13

Soft 48.0 30.0 30.0 2.0 7.2-7.6 40-48 30-35

Mod. Hard 96.0 60.0 60.0 4.0 7.4-7.8 80-100 60-70

Hard 192.0 120.0 120.0 8.0 7.6-8.0 160-180 110-120

Very Hard 384.0 240.0 240.0 16.0 8.0-8.4 280-320 225-245

a Approximate equilibrium pH after 24-hour aeration

b Expressed as mg/L CaCO3

When standard laboratory-reconstituted water is cited as the dilution water, and no additional

measurements are reported, the recommended approach for estimating ion concentrations is to use

the ion concentrations calculated from the amount of salts added for the type of reconstituted water

reported in the article. For example, if the range of hardness of the reconstituted water is reported as

80-100 mg/L CaCO3, then the specific ion concentrations calculated from the standard recipe for

moderately hard reconstituted water should be used for BLM input (see Table 2 and example

calculation in Appendix D-2). The use of ion concentrations calculated from the standard recipes

assumes that salts were stored in a manner to prevent hydration and that technician errors in

weighing of salts, measurements of dilution water, and measurement of solution volumes were

minimal. 

Alternatively, if the authors state that moderately hard water was prepared following one of the

standard recipes, and they measured the hardness of the water, then the calculated ion concentrations

should be adjusted to account for any difference from the mean of the expected range. For example,

if the mean measured hardness in a test water prepared using the recipe for moderately hard

reconstituted water was 78 mg/L CaCO3, the Ca:Mg ratio would be 0.700 for all reconstituted water

types, and the respective Ca and Mg concentrations could be calculated using the following equations:

Ca = (0.4008 × measured hardness)÷[1+(1÷Ca:Mg ratio)] Equation 1

Mg = (0.2431 × measured hardness)÷(1+Ca:Mg ratio) Equation 2

The remaining ion concentrations are each multiplied by 0.92 (quotient of 78 and 85 mg/L CaCO3,

the latter of which is the expected hardness for moderately hard reconstituted water), as in Table 1.
C-6

Table 3 provides ion concentrations predicted for a standard reconstituted water mix using the

hardness adjustment in accordance with the example above.

Note that this same rationale for scaling the default major anions and cations in reconstituted

water also applies to a variety of natural surface and well waters. Analysis of St. Louis River, MN,

water and Western Fish Toxicology Station (WFTS) well water indicated that a strong linear

relationship also exists between water hardness and the major anion (Cl, SO4) and cation (Ca, Mg,

Na) concentrations in these water types (see Sections 2.6, 2.7, and 2.19). The strong relationships

are consistent with findings 

Table 2. Calculated Ion Concentrations Based on the Standard Salts Added

Water Type

(Nominal Hardness Range)

Specific Ionsa (mg/L)

Ca:Mgb

Expected Hardness

(mg/L CaCO3) Ca Mg Na K Cl SO c 4

Very Soft

(10-13 mg/L CaCO3)

1.75 1.51 3.28 0.262 0.238 10.2 0.700 11

Soft

(40-48 mg/L CaCO3)

6.99 6.06 13.1 1.05 0.951 40.7 0.700 42

Moderately Hard

(80-100 mg/L CaCO3)

14.0 12.1 26.3 2.10 1.90 81.4 0.700 85

Hard

(160-180 mg/L CaCO3)

27.9 24.2 52.5 4.20 3.80 163 0.700 170

Very Hard

(280-320 mg/L CaCO3)

55.9 48.5 105 8.39 7.61 325 0.700 339

a Ion concentrations were calculated from standard salt recipes (refer to Table 1 and example calculation for very soft

water in Appendix D-1).

b Ratio equals quotient of (Ca÷40.08) and (Mg÷24.31), where 40.08 and 24.31 are the molecular weights of Ca and

Mg, respectively, in units of mg/mmol.

c Hardness calculated according to the concentrations of Ca and Mg given here and the equation given in Appendix

D-1.

Table 3. Adjusted Ion Concentrations for a Standard Reconstituted Water Mix Based on 

Reported Hardness

Moderately Hard Reconstituted

Water

Hardness

(mg/L CaCO3)

Specific Ions (mg/L)

Ca Mg Na K Cl SO4

Nominal 85a 14.0 12.1 26.3 2.10 1.90 81.4

Adjusted 78 12.9 11.2 24.2 2.10 1.75 74.9

a Expected hardness based on the amount of salts added (from Table 1). Calcium and magnesium are calculated

using Equations 1 and 2. Other adjusted values (italic and bold) are a result of the product of the ratio of measured

hardness (78 mg/L) to expected hardness (85 mg/L) and nominal ion concentrations, e.g., the adjusted sodium ion

concentration for a standard laboratory reconstituted water mix based on a reported total hardness of 78 mg/L CaCO3

is: 78÷85=0.92; 0.92*26.3=24.2.
C-7

presented in an earlier comprehensive report by Erickson (1985). Note, however, that because there

is generally poor correlation between K and water hardness in the various ambient surface and ground

water types (see Section 2.6), the value calculated for K should not be scaled according to hardness.

2.2 pH Adjustment with HCl

Schubauer-Berigan et al. (1993) adjusted pH using HCl but reported only nominal hardness and

alkalinity. The tests were conducted at the EPA Office of Research and Development, MidContinent Ecology Division, Duluth, MN, using a standard very hard reconstituted water mix. The

authors need to be contacted to obtain any additional water chemistry data they might have.

Recommendation: Alkalinity and hardness were not measured in the tests reported in

Schubauer-Berigan et al. (1993), and no additional water chemistry data are available from the study

(Phil Monson, U.S. EPA-Duluth, personal communication). The HCl required to adjust the pH was

assumed to be added in amounts too small to significantly affect any of the other water quality

parameters (Gerald Ankley, U.S. EPA-Duluth, personal communication). Based on these remarks, we

believe ion concentrations for this particular study should be estimated using methods outlined in

Section 2.1. 

2.3 Estimation of DOC

How should DOC be estimated if only total organic carbon (TOC) was measured in the study?

Can DOC be estimated if no measurements of organic carbon were reported in the study?

Recommendation: As a general rule, TOC values can be used directly in place of DOC for

dechlorinated and de-ionized city tap water, well water, and oligotrophic lake water (e.g., Lake

Superior water). TOC values are not recommended in place of DOC for water from estuaries,

wetlands, or higher order streams unless data are included that indicate otherwise. Rather, the

proportion of organic carbon expected to be dissolved in surface waters should be estimated and used

to scale the measured TOC value. When possible, the DOC:TOC ratio for a surface water should be

obtained using the USGS NASQAN dataset. The NASQAN dataset can be reached through the USGS

Web site (water.usgs.gov/nasqan/data/finaldata.html). If a representative ratio for a particular body of

water cannot be determined, the ratio for the particular water type (lake or stream) should be

obtained from the final draft of the Ambient Water Quality Criteria Derivation Methodology Human

Health Technical Support Document (U.S. EPA 1998a, Table 2.4.11). A summary of these data, by

State, is provided in Appendix D-2. In this appendix, TOC is operationally defined as the sum of

DOC and particulate organic carbon (POC). The national mean fraction of organic carbon is 86

percent for streams and 88 percent for lakes. The DOC:TOC ratio can be applied to lakes or streams

within a State to obtain an estimate of DOC from values reported for TOC.

Example:

Reference Water Body TOC (mg/L) DOC:TOC Estimated DOC (mg/L)

Lind et al. manuscript St. Louis R, MN 32 0.87 28
C-8

For tests with reconstituted, city tap, or well water, default DOC values can be applied if the

author does not report a measured value. The recommended default TOC (DOC) value for laboratory

prepared reconstituted water is 0.5 mg carbon/L (note: some newer laboratory water systems can

achieve a TOC of less than 0.5 mg/L). For regular city tap and well water, a value of 1.6 mg carbon/L

can be assumed. The recommended default value for laboratory-prepared reconstituted water is based

on the arithmetic mean of recent measurements of DOC in reconstituted water prepared at two

Federal (U.S. EPA Cincinnati, OH, and USGS Yankton, SD) and two consulting (Commonwealth

Biomonitoring and GLEC) laboratories (range 0.1 to 1 mg/L). The recommended default value for

dechlorinated city tap and well water is based on the arithmetic mean of measurements of DOC in

source water from Lake Ontario (Environment Canada, Burlington, ON) and the New River, VA

(City of Blacksburg, VA), and well water from Oak Ridge National Laboratory (Oak Ridge, TN) and

EPA’s WFTS (Corvallis, OR). The DOC values in these waters ranged from 1.1 to 2.5 mg/L.

For tests conducted in surface waters, we do not recommend the use of a default DOC value

because of the large variability of DOC observed. Rather, a reliable database such as USGS NASQAN

(as described above) should be searched for DOC measurements. If a database such as NASQAN is

consulted, only those DOC measurements closest to the time of the study should be considered as

surrogate values. In general, these DOC concentrations should not differ by more than a factor of

1.25. If DOC measurements for the surface water cannot be obtained from a reliable source, then the

toxicity test should not be included in Table 1 for BLM normalization.

2.4 DOC in Lake Superior Water

Lake Superior water has been used in a number of acute and chronic toxicity studies included in

the Aquatic Life Criteria for Copper (U.S. EPA 1998b). Dissolved organic matter (DOM) in Lake

Superior is assumed to be anywhere from 1 to 3 mg/L (Russ Erickson, U.S. EPA-Duluth, personal

communication; McGeer et al. 2000). This value is expected to be at least 90 percent of TOC (or 2

mg/L) (see Spehar and Fiandt 1986). A default value based on recent measurements is needed for

DOC in Lake Superior water.

Recommendation: Recent measurements of TOC in Lake Superior dilution water are in

Appendix D-3 (Greg Lien, U.S. EPA-Duluth, personal communication). The geometric mean

concentration of TOC in Lake Superior dilution water from multiple measurements is 1.27 mg/L.

Given the recommendation in Section 2.3, the recommended DOC for Lake Superior dilution water is

1.1 mg/L (1.27 mg/L × 0.88). 

2.5 Applying Water Chemistry Data to Lake Superior Water

The ionic composition included in the Table 1 spreadsheet for Lake Superior water is based on

concentrations converted from values reported in Erickson et al. (1996b): Ca at 0.68 meq/L = 13.6

mg/L; Mg at 0.24 meq/L = 2.9 mg/L; Na at 0.065 meq/L = 1.5 mg/L; K at 0.015 meq/L = 0.59 mg/L;

SO4 at 0.070 meq/L = 3.4 mg/L; Cl at 0.035 meq/L = 1.2 mg/L; and alkalinity at 0.85 meq/L = 43

mg/L. The concentrations for most of these parameters were also reported in Biesinger and

Christensen (1972) and approximate those listed above. Should the Erickson et al. (1996b) data be

applied to all Lake Superior studies, or is there a stronger rationale for applying the Biesinger and

Christensen (1972) data to the older studies?
C-9

Recommendation: We recommend applying the mean of the Erickson et al. (1996b) citation

and Biesinger and Christensen (1972) water chemistry data to all Lake Superior studies prior to 1987,

when the results were initially reported. After 1987, we recommend use of the Erickson et al.

(1996b) water chemistry data alone (Table 4). For each test, Ca and Mg concentrations should be

estimated using Equations 1 and 2, the Ca:Mg ratios given below, and the measured hardness of the

test water (Section 2.1). Ions other than K should be scaled according to the measured test hardness,

also discussed in Section 2.1.

Table 4. Recommended Spreadsheet Addition for Lake Superior Dilution Water

Applied to:

Hardness

(mg/L CaCO3)

Alkalinity

(mg/L CaCO3)

Specific Ions (mg/L)

Ca Mg Ca:Mg Na K Cl SO4

Pre-1987a 46 42 13.6 3.0 2.75 1.3 0.57 1.2 3.4

Post-1987b 46 43 13.6 2.9 2.84 1.5 0.59 1.2 3.4

a Mean of the Erickson et al. (1996b) and Biesinger and Christensen (1972) water chemistry data

b Erickson et al. (1996b) water chemistry data alone

2.6 Predicting Ionic Composition of WFTS Well Water

The following studies seem were conducted at EPA’s WFTS using well water: Andros and

Garton (1980), Chapman (1975, 1978), Chapman and Stevens (1978), Lorz and McPherson (1976),

Nebeker et al. (1984a, 1986a, b), and Seim et al. (1984). Among these studies, however, there is a

wide range of hardness values (20-100 mg/L), and the ionic composition of the water was not always

reported. 

The large variation in WFTS well water hardness, and consequently, ionic composition, is due to

seasonal variability (Samuelson 1976). The TOC content of this water has been reported to be 1.1

mg/L (McCrady and Chapman 1979), of which 100 percent is expected to be dissolved. A general

strategy is needed to predict the ionic composition of WFTS well water based on measured water

hardness. 

Recommendation: The well feeding the WFTS is susceptible to influx from ground water

during rain events in late fall and winter (November through March or April). During this period the

water 

hardness can reach measured levels as high as 100 mg/L CaCO3. Over the remaining months

(particularly from July to November), hardness stabilizes at around 25 to 40 mg/L CaCO3, as do other

water quality parameters (Al Nebeker, U.S. EPA Corvallis, personal communication; Samuelson

1976). It is important to note that the high hardness reported for WFTS well water is sporadic, even

in the winter.

The recommended strategy for filling the existing gaps in data reported from studies using this

well water is to estimate the ion concentrations on the basis of their relationship to the total

hardness measured during a particular test. The acceptability of tests conducted using WFTS water

depends on the range of hardness values reported, i.e., if the hardness varies widely over the course of

a particular test, then perhaps the test should not be used. Regression analyses were performed using

measured hardness and ion data for the WFTS well water reported in Samuelson (1976), April 1972
C-10

to April 1974, and supplemented with additional data from Gary Chapman, personal communication

(only those data from May 1974 to April 1978; see Appendix D-4). These relationships and the

corresponding regression equations are presented in Figures 1 through 6 (found at the end of this

report). Major ion concentrations for WFTS well water were predicted using the regression equations

over a wide range of water hardness (10 to 80 mg/L CaCO3) to determine the accuracy of the

procedure (Table 5). The error between predicted and measured ion concentrations is generally within

10 percent for all ions except K, where a default value of 0.7 mg/L was chosen for all hardness levels

(actual range is 0.1 to 1.1 mg/L, with the majority of data falling between 0.5 and 0.9 mg/L). The

correlation coefficient (R2) for the relationship between K and water hardness in WFTS well water

was only 0.124. Note: BLM predictions of copper gill accumulation and toxicity are relatively

insensitive to the concentration of K, so errors in its estimation should not appreciably affect model

predictions. The following regression equations were used to generate the example data provided in

Table 5:

[Ca] = 0.3085 + (measured hardness * 0.2738)

[Mg] = 0.5429 + (measured hardness * 0.0573)

[Na] = 3.3029 + (measured hardness * 0.0713)

[Cl] = 2.7842 + (measured hardness * 0.1278)

[SO4] = -3.043 + (measured hardness * 0.2816)

Lorz and McPherson (1976) and the Seim et al. (1984) tests were not run in WFTS well water,

but in water from different wells along the Willamette River. Water chemistry appears to be less

variable for these wells (Harold Lorz and Wayne Seim, personal communication). The following

additional water chemistry information for the two well water types used in these studies was

provided by the respective authors in January 2001.

Many of the studies conducted by Chapman used reverse osmosis treatment to maintain a

blended water supply that was of essentially constant ion content throughout the tests. All the test

data from Chapman appear to be acceptable; the only test complicated by fluctuating hardness was

the 22-month chronic zinc test with sockeye salmon, and that test produced only a NOEC.

Table 5. Predicted Ion Concentrations in WFTS Well Water Based on Measured

Hardness

Total Hardness

(Mean Measured value)

mg/L CaCO3

Predicted Ion Concentrations (mg/L)

Ca Mg Na Cl SO4

Defaulta

K

15.00 4.42 1.40 4.10 4.70 1.18 0.70

20.00 5.78 1.69 4.46 5.34 2.59 0.70

25.00 7.15 1.98 4.82 5.98 4.00 0.70

30.00 8.52 2.26 5.17 6.62 5.41 0.70

35.00 9.89 2.55 5.53 7.26 6.81 0.70

40.00 11.26 2.83 5.88 7.90 8.22 0.70

45.00 12.63 3.12 6.24 8.54 9.63 0.70

50.00 14.00 3.41 6.60 9.17 11.04 0.70

55.00 15.37 3.69 6.95 9.81 12.45 0.70

60.00 16.74 3.98 7.31 10.45 13.85 0.70

65.00 18.11 4.27 7.67 11.09 15.26 0.70
C-11

70.00 19.47 4.55 8.02 11.73 16.67 0.70

75.00 20.84 4.84 8.38 12.37 18.08 0.70

80.00 22.21 5.13 8.74 13.01 19.49 0.70

a Value not corrected. Assume default value of 0.70 mg/L.

Recommended Spreadsheet Addition for Oregon Well Water.

Applied to:

Hardness

(mg/L

CaCO3)

Alkalinity

(mg/L

CaCO3) pH DOC

Specific Ionsa (mg/L)

Ca Mg Ca:Mg Na K Cl SO4

Lorz and

McPherson

1976

95 66 6.8-7.9 1.6B 19 12 1.0 7.6 1.0 7.0 12

Seim et al.

1984

120 126 7.7 1.6B 34 8.6 2.4 15 0.7 5.0 2.3

a Specific ion values were obtained through personal communication with the primary authors; hardness, alkalinity,

and pH values are as reported in the article. The Ca:Mg ratios were calculated on the basis of data provided by

authors, then Ca and Mg values used were back-calculated on the basis of these ratios and the measured test

hardness (see Equations 1 and 2).

b Suggested default value for untreated well water (see Section 2.3).

2.7 Data for Measurement of Blacksburg/New River Water

A substantial amount of acute copper toxicity data to various freshwater organisms is reported

using dechlorinated City of Blacksburg, VA, tap water. These include studies by Belanger et al.

(1989), Cairns et al. (1981), Hartwell et al. (1989), and Thompson et al. (1980). Hardness,

alkalinity, and pH values are reported for City of Blacksburg water in all of these studies, but the

ionic compositional data are not. This information is required to obtain BLM-normalized LC50s for

these data.

Recommendation: According to Don Cherry (personal communication), tests conducted at

Virginia Polytechnic Institute and State University used City of Blacksburg, VA, tap water, which is

drawn from the nearby New River. Don Cherry collected a sample of New River water for analysis

under Work Assignment 1-20. The results of the analysis are provided in Appendix D-5. The sample

was of untreated natural water prior to any treatment by the City of Blacksburg. Values for treated

New River water (city) were provided by Jerry Higgins, Water Superintendent, City of Blacksburg.

Table 6 summarizes the measured values for New River and City of Blacksburg dechlorinated tap

water.

Historically, hardness and alkalinity vary substantially in dechlorinated City of Blacksburg tap

water and in raw New River water (Table 6). Some of this difference may be attributed to seasonal

effects. For example, strong seasonal influence was observed in both well water (influenced by surface

water, i.e., WFTS well water; see Section 2.6) and a natural surface water (St. Louis River, MN; refer

ahead to Section 2.19). Previously, we plotted ion concentrations against hardness for each of these

two water types (Figures 1 through 6 and Appendix D-6). The relationships were good in almost all

cases (positive, R2 = 0.5 to 0.9), and the resultant regression equations were used to scale ion

concentrations according to reported water hardness. Incomplete datasets, however, preclude the use
C-12

of the same approach for City of Blacksburg tap and raw New River water. Instead, we recommend

using the ion and hardness values from the City of Blacksburg water sample and USGS NASQAN ion

data, respectively (Table 6), to generate surrogate ion values for the respective waters that were not

reported in the previous studies (indicated by the shaded area in Table 6). The operation is simply to

multiply ion concentrations for the “ acquired data” by the ratio of hardness values in City of

Blacksburg and NASQAN water and the corresponding test waters as was done in Section 2.1. We used

the NASQAN ion data as the basis for scaling the raw New River water ion estimates because

NASQAN represents data collected over several representative years, including the years in the

timeframe in which the studies of interest were initiated and completed. The exception was with

DOC. We felt that the DOC value obtained from the sample of New River water collected in August

2000 would be more representative than the few values generated from NASQAN (all pre-1980).

2.8 Cu Concentrations and Alkalinity

The methods sections of both Belanger and Cherry (1990) and Belanger et al. (1989) state that

total and dissolved Cu were measured, but it is not clear whether the reported LC50s are based on

total or dissolved copper concentration. Also, in Belanger and Cherry (1990), pH was adjusted with

sodium hydroxide (NaOH) or nitric acid (HNO3), but only nominal pHs were reported. Alkalinity and

hardness after pH adjustment were not reported. Can alkalinity be adjusted for these tests?

Recommendation: The concentration Cu in algae is reported on a total metal basis in

Belanger et al. (1989) and Belanger and Cherry (1990). The Cu in water is reported on an acidsoluble basis. The acid-soluble concentration of Cu in water was used to derive the LC50. For all

intents and purposes, acid-soluble Cu can be considered as dissolved Cu because the acidification of the

filtrate after filtration is probably sufficient to obtain most of the Cu associated with colloidal

material. Normally a digestion procedure is required to convert all Cu to the dissolved form. If the

sample had not been filtered, it would not have been acceptable because it could have been elevated

by dissolution of particulate copper.

The pH levels achieved in the batch culture pH tests in Belanger and Cherry (1990) were

reported as 6.15, 8.02, and 8.95. Given the proximity of these values to the desired target pH values

of 6, 8, and 9, respectively, it would appear that the researchers were able to closely approximate the

nominal pH levels, including those selected for the acute heavy metal tests (also pH 6, 8, and 9,

respectively). Assuming that the target pH values of 6, 8, and 9 were achieved in the acute tests,

adjustment with NaOH and HNO3 would have affected alkalinity, but probably not hardness or the

major anion and cation concentrations, except possibly Na. The contribution to Na by the addition

of NaOH was probably small, so no further adjustment would be necessary. 
C-13

Table 6. Comparison of Values for Untreated (Natural) and Treated (Dechlorinated City of Blacksburg, VA) New River Wat

Source

Water

Type pH

Total

Hardness

(mg/L CaCO3)

Total

Alkalinity

(mg/L CaCO3)

Specific Ions (mg/L) 

Ca Mg Na K Cl SO4 

Acquired Data

City of Blacksburg, VAa City 8.5 44 39 - -9.3 -33 45

Cherry 2000 (08/00)b New R. 8.0 - 52 15 0.6 6.6 2.0 6.1 9.8 

NASQANc New R. - 61 - 15 5.8 3.4 1.6 4.0 13 

 Values To Be Applied to Table 1 Toxicity Testsd

Belanger et al. 1989 City 7.7 45 40 11 4.2 9.5 1.6 34 46 Hartwell et al. 1989 City 7.5 72 43 18 6.8 15 1.6 54 74 Cairns et al. 1981 City 7.0 26 27 6.4 2.4 5.5 1.6 19 26 Thompson et al. 1980 City 7.2 40 28 9.9 3.8 8.5 1.6 30 41 Belanger et al. 1989 New R. 8.2 94 70 23 8.8 5.2 1.6 6.2 20 Belanger and Cherry 1990 New R. 6, 8, 9 98 74 24 9.1 5.4 1.6 6.4 21 

a Data provided by Gerard (Jerry) Higgins of Blacksburg-Christianburg VPI Water Authority, Blacksburg, VA. Values presented are from a gra

collected January 31, 2000. Organic carbon (originally measured and reported as TOC) is assumed to be 100 percent dissolved.

b Sample provided by Don Cherry, Virginia Polytechnic Institute and State University, Blacksburg, VA, and analyzed by Environmental Heal

South Bend, IN. Values presented are from a grab sample collected August 2000. The value for Mg of 0.6 mg/L appears to be a reporting error, anused for subsequent calculations of total hardness or scaling of ion values.

c Data obtained from USGS NASQAN database. Values presented are means of 213 samples, except for DOC, which is a mean of seven samples, 

analyzed from January 1973 to August 1995.

d Shaded area indicates mean values estimated from previously (NASQAN) or recently measured (Cherry 2000 or City of Blacksburg; nonadjus

All values have been rounded to two significant figures. Shaded values were derived according to text above using the approach outlined in S
C-14

Using a nomograph found in Faust and Aly (1981), alkalinity at pH 6 should be approximately

33 percent of the alkalinity at pH 8, and alkalinity at pH 9 should be 5 percent higher than the

alkalinity at pH 8 (Table 7). Therefore, the values for alkalinity in Table 7 should be used for the

acute toxicity tests presented in Belanger and Cherry (1990) in this case. For other analyses,

different adjustment factors may be appropriate, based on other interpretations from the Faust and

Aly nomograph or other methods as well. Appropriate consideration should also be given to the test

system equilibration with the atmosphere. 

Table 7. Estimated Alkalinity in Natural Surface Water Based on pH

Source Water Nominal pH Alkalinity (mg/L CaCO3)

New River 6 24.5

8.1 74.2a

9 77.9

Clinch River 6 47.6

8.3 144a

9 152

Amy Bayou 6 40.2

8.3 122a

9 128

a Indicates values reported in text.

2.9 Calculation of DOC and Humic Acid

What was the technical approach used to calculate DOC and percent humic acid (HA) for the

Winner (1985) toxicity tests?

Recommendation: At a nominal HA concentration of 0.0 mg/L in soft and medium hardness

test waters, the DOC is assumed to be that of the ultrapure laboratory water, which is estimated to be

0.3 mg/L (approximately one-half of the recommended default value for DOC in laboratory water;

see Section 2.3). At nominal HA concentrations of 0.15, 0.75, and 1.50 mg/L, the DOC is calculated

by dividing by a value of 2, based on the assumption in the BLM User’s Guide (Di Toro et al. 2000)

that the percent carbon in HA is 0.50 (see example below and Table 8). Because the water used to

obtain these HA concentrations was ultrapure laboratory water, 0.3 mg carbon/L was added; final

rounded values of 0.38, 0.68, and 1.1 are recommended.

Table 8. Estimates of Dissolved Organic Carbon and Percent Humic Acid for the Winner

(1985) Toxicity Tests

Humic Acid Added (mg/L)a Calculated DOC (mg/L) Calculated Percent Humic Acid

0 0.3 10

0.15 0.38 28

0.75 0.68 60

1.5 1.1 74
C-15

a As indicated in Table 3 of Winner (1985).

2.10 Alkalinity of Lake Superior Water

For the Lind et al. (manuscript) tests conducted in Lake Superior water (adjusted with CaSO4 or

MgSO4), is there any way to estimate alkalinity values?

Recommendation: For tests conducted in Lake Superior water, assume an alkalinity of 42

mg/L CaCO3 (see Section 2.5).

2.11 Availability of LC50s

The LC50s reported by Collyard et al. (1994) are shown graphically in publication. The LC50s

provided in Table 1 are interpolated from the figure. Are the actual measured LC50s available from

the authors?

Recommendation: The actual LC50s generated and presented graphically in Collyard et al.

(1994) have been archived at U.S. EPA-Duluth, as reported by Gerald Ankley (personal

communication, 3 November 2000). These values are not readily available in any other form. The

data are acceptable as is on the basis of recommendations in the Guidelines (Stephan et al. 1985).

Precedence for the use of values gleaned from graphical data is provided in the 2001 Update of

Ambient Water Quality Criteria for Cadmium (U.S. EPA 2001). 

2.12 Cl and Na Concentrations

Cl and Na ion concentrations of the tap water used for testing in Rice and Harrison (1983) were

derived from the addition of 20 mg/L sodium chloride (NaCl). What are the specific concentrations

of the individual ions from the addition of the salt? What concentrations do you suggest using for K

and SO4 in this water?

Recommendation: The Cl content of the tap dilution water used in Rice and Harrison (1983)

was reported as having been derived from the addition of 20 mg/L of NaCl. Assuming that the initial

Na and Cl concentrations in tap water were essentially zero, the concentrations of these ions can be

calculated in the following way:

The molecular weight of NaCl is 58.44 g/mol. The atomic weight of Na is 22.98 mg/L and the

atomic weight of Cl is 35.453 mg/L.

The concentration of Na is:

20 mg NaCl/L * 1 mmol NaCl/58.44 mg NaCl = 0.342 mmol NaCl/L.

0.342 mmol NaCl * 1 mmol Na/1 mmol NaCl * 22.98 mg Na/1 mmol Na

= 7.86 mg Na/L.

The concentration of Cl is:

20 mg NaCl/L × 1 mmol NaCl/58.44 mg NaCl = 0.342 mmol NaCl/L. 
C-16

0.342 mmol NaCl × 1 mmol Na/1 mmol NaCl × 35.453 mg Cl/1 mmol Cl

= 12.12 mg Cl/L.

Given the potentially large dichotomy between the default ion concentrations and measured

hardness of the water used in this study, we recommend adjusting the default SO4 concentration

according to measured hardness as in Section 2.1. We do not, however, recommend adjusting the

current default value of 1.0 mg/L for K.

2.13 Calculating DOC in Dilution Water

The dilution water used in the acute copper toxicity tests with cutthroat trout in Chakoumakos

et al. (1979) was a different mix of spring water and de-ionized water for each test. Ca and Mg

concentrations were measured and reported for each of the test waters used, but measurements of the

other ions were reported only for the undiluted spring water. Based on a percentage dilution, ions

other than Ca and Mg were estimated in the following way: hardness was measured in the spring water

and in each of the test waters; the proportion of spring water was calculated for each test using these

measured hardness values; this proportion was then multiplied by the concentration of, for example,

Na in the spring water to get an estimated Na value for each test. TOC in the spring water was 3.3

mg/L. Should the same approach as that used to estimate the other ions be used to calculate DOC,

which was only measured in undiluted spring water? 

Recommendation: The concentrations of the major cations and anions in the dilution water

used by Chakoumakos et al. (1979) were calculated based on the percent dilution of natural spring

water with de-ionized water. The same correction can be used to estimate DOC, with the following

assumptions. First, the TOC in spring water was 100 percent dissolved. Second, the DOC of deionized water was 0.5 mg/L. If these assumptions are acceptable, the DOCs for H/H, M/H, L/H, H/M,

M/M, L/M, H/L, M/L, and L/L would be 3.3, 1.5, 0.75, 3.3, 1.7, 0.94, 2.8, 1.5, and 0.87 mg/L,

respectively.

2.14 Ionic Composition of Chehalis River Water

The ionic composition of Chehalis River, WA, water is needed to fill in existing data gaps used

for BLM analysis of acute toxicity reported in Mudge et al. (1993). The publication states, “ Water

quality data collected during this bioassay program is similar to historical data for Chehalis River

(WPPSS 1982) and other Pacific NW streams (Samuelson 1976).” Are data from Samuelson (1976)

acceptable for use in approximating these ion concentrations? Furthermore, are there any dissolved

or ionic LC50s available other than those reported in the publication?

Recommendation: The following additional water chemistry information for the Chehalis

River dilution water used in the studies reported by Mudge et al. (1993) was provided by the author on

20 November 2000. These measurements were made on Chehalis River water at the time of testing.

A corresponding value for DOC was obtained from the NASQAN dataset.

Recommended spreadsheet addition for Chehalis River dilution water

Applied to:

DOC

(mg/L)

Specific Ions (mg/L)

Ca Mg Ca:Mg Na K Cl SO4
C-17

Mudge et al. 1993 3.2a 7.1 2.4 1.8 5.1 0.65 4.5 (May)

4.2 (Jun)

3.1 (Sep)

4.0 (May)

3.5 (May-Jul)

2.3 (Sep)

a Value from the USGS NASQAN dataset, 1980-1982, when the tests were conducted.

2.15 Chemistry of Water in Howarth and Sprague (1978)

What is the ionic composition and organic carbon content of test waters used in Howarth and

Sprague (1978)? The waters used for testing were various mixes of University of Guelph (Guelph,

ON, Canada) well water and de-ionized well water. The de-ionized well water was reported as “ having

retained its original chloride content (22 mg/l),” but the values for the other major anion and cation

concentrations were not reported. Furthermore, the equation provided for calculating alkalinity from

pH and hardness (supposedly accounting for 96.7 percent of the variability) appears unreliable. For

example, using the equation and a total water hardness of 364 mg/L CaCO3 at pH 9, one obtains an

estimated alkalinity value of 341 mg/L CaCO3. In contrast, the measured alkalinity reported in the

text for this level of hardness and pH was 263 mg/L CaCO3.

Recommendation: The equation provided in the text of Howarth and Sprague (1978) for

calculating alkalinity appears unreliable. The calculated alkalinity does not approximate measured

alkalinity within a reasonable degree of accuracy. Values of hardness, pH, and alkalinity in Dixon and

Sprague (1981a), which used the same water source in their toxicity tests, give greater evidence of

this; i.e., using the measured value of hardness of 374 mg/L CaCO3 and a pH of 7.75, the alkalinity

calculated with the equation is 98 mg/L CaCO3. This compares rather poorly with the measured

alkalinity of 223 mg/L CaCO3. Instead, alkalinity can be estimated using the nomograph from Faust

and Aly (1981) as in Section 2.8.

It is possible to apply the procedure used with the Chakoumakos et al. (1979) data here, i.e.,

using the ratio of hardness in full-strength well water and de-ionized well water to calculate the

dilution of the other major ion concentrations. However, no values are given for Na or K in

University of Guelph well water. This study is also complicated by the reverse-osmosis unit used to

create the de-ionized well water. In particular, the statement concerning the retention of the original

Cl concentration in the de-ionized well water implies an ionic exchange that would also require a

cation (to maintain charge balance). The cation involved is unknown. As discussed in a phone

conversation with John Sprague on 17 November 2000, and later that day with Scott Howarth

(Environment Canada), NaCl may have leached through the RO unit. Assuming that Na and Cl

leached through the unit in equivalent proportions, a value of 14 mg/L for Na can be back-calculated

from the reported Cl concentration of 22 mg/L.

Default DOC concentrations of 1.6 and 0.5 mg/L were assumed for the well water and deionized water used in the tests, respectively (see Section 2.3). The DOC concentrations were adjusted

for each particular test water hardness level based on the proportion of well water and de-ionized

water used to achieve the desired test hardness level. In the example provided in Table 9, the dilution

factor of 0.27, based on the ratio of the average hardness of well water (366 mg/L CaCO3) versus the

average hardness of well plus de-ionized well water (100 mg/L CaCO3), was applied to the starting

DOC concentrations to achieve an estimate of the DOC concentrations at 100 mg/L CaCO3). Table
C-18

9 shows the results of similar adjustments made for the major anions and cations based on the data

reported in Howarth and Sprague (1978).

2.16 Default Values for Analyte Concentrations

What value should be used when a specific analyte is not detected at its designated detection

limit?

Recommendation: The use of half the detection limit (DL) is most appropriate when the

concentration of an analyte is not detected. One-half the DL will closely approximate a replacement

value for censored data in a log-normally distributed population that includes several measured values

(Berthouex and Brown 1994; Dolan and El-Shaarawi 1991). This way some of the “ nondetect”

samples will actually be counted as detected.

Table 9. Example Calculations to Estimate Water Chemistry of Tests Conducted at 100

mg/L CaCO3 by Howarth and Sprague (1978) Using a Mixture of University of Guelph

Well Water and De-ionized Water

Parameter 

(units in mg/L) De-ionized water Well Water 

Example Calculations

for Mixture

Hardness 0 366 100

(i.e., 0.27 dilution factor)

Ca 0 77 (from Dixon & Sprague

1981)

21

Mg 0 43 (from Dixon & Sprague

1981)

12

Na 14 (assuming NaCl used for

the softening process)

14 (estimated from [Cl]) 14

K 0 2.4 (based on personal

communication from Dr. Patricia

Wright, Univ. of Guelph,

Guelph, ON)

0.66

Cl 22 (stated as not having

changed from the water

softening process)

22 22

SO4 0 129 35

DOC 0.5 (default value for deionized waters)1.6 (default value for well

waters)

0.8

Alkalinity (calculated using ratios as in Section 2.8):

at pH 6 0a 81.5 22

at pH 7 0a 205 55

at pH 8 0a 250 N/A

at pH 9 0a 263 70

a Alkalinity in de-ionized well water is assumed to be 0.0 mg/L.
C-19

2.17 Organic Carbon Content of Samples

Can any information be obtained on the organic carbon content of the spring water / City of

Cincinnati, OH, tap water mixes used in Brungs et al. (1973), Geckler et al. (1976), Horning and

Neiheisel (1979), Mount (1968), Mount and Stephan (1969), and Pickering et al. (1977)?

Recommendation: The water used for all tests was a mixture of spring-fed pond water

(originating at the Newtown Fish Farm) and carbon-filtered, demineralized Cincinnati tap water. The

water was mixed to achieve the desired test hardness level and discharged to a large (several thousand

gallon) concrete reservoir that fed the test system. The detention time varied anywhere from 30 to

90 days, depending on the study, which was sufficient to allow the growth of phytoplankton and

zooplankton in moderate abundance. No additional information regarding the TOC (DOC)

concentration or treatment of this water is available at this time. The recommended organic carbon

content of spring/city water mix is currently a conservative 1.6 mg/L, but could be as high as 2.5

mg/L, the highest DOC concentration recorded for a natural surface or well water used for studies

included in this report (see Section 2.3). Considering the long retention time, and the fact that the

natural water was spring-fed pond water, the more conservative DOC value of 2.5 mg/L is

recommended for this water.

2.18 Additional Water Chemistry Data Needed

Additional water chemistry data are needed for Bennett et al. (1995) and Richards and Beitinger

(1995). In the case of Richards and Beitinger 1995, only the ranges of measured pH, alkalinity, and

hardness across all tests were given. 

Recommendation: Detailed pH, alkalinity, and hardness values were provided by both

Bennett et al. (1995) and Richards and Beitinger (1995) (Appendixes D-7 and D-9, respectively).

The studies performed by Bennett et al. were conducted using dechlorinated City of Denton, TX, tap

water (from Lake Roy Roberts). The author was not able to provide any additional data regarding the

ionic composition of this water; however, based on supplementary data, mean values of pH,

alkalinity, and temperature were 8.07 and 89.7 mg/L CaCO3 and 21.4 C, respectively. Richards and

Beitinger’s studies were conducted using standard reconstituted (hard) water. To estimate the ionic

composition of this water, refer to recommendations provided in Section 2.1.

2.19 Estimating Data for Waters

Values for DOC, TSS, Ca, Mg, Na, K, SO4, and Cl are needed for the following natural waters:

Water Body Reference

American River, California – sand filtered Finlayson and Verrue 1982

Clinch River – 11:m filtered Belanger et al. 1989

Belanger and Cherry 1990

Amy Bayou Belanger and Cherry 1990

Blaine Creek, Kentucky – 1.6 :m filtered Dobbs et al. 1994

S. Kawishiwi Lind et al. manuscript

St. Louis River Lind et al. manuscript

Lake One Lind et al. manuscript
C-20

Colby Lake Lind et al. manuscript

Cloquet Lake Lind et al. manuscript

Greenwood Lake Lind et al. manuscript

Embarrass River Lind et al. manuscript

Green Duwamish River Buckley 1983

Chehalis River Mudge et al. 1993

Pinto Creek, AZ Lewis 1978

Naugatuck River Carlson et al. 1986

Recommendation: On the following pages are data (current and/or historical, presented as

arithmetic means) from selected natural waters that were retrieved from NASQAN, STORET, or a

secondary source (as indicated). As mentioned earlier (see Sections 2.6 and 2.7), given the reasonably

good correlation between most of the major anion and cations (except K) and water hardness in

natural surface and well waters, we recommend using the ion and hardness values retrieved from these

various sources to estimate the ion concentrations in the test water used in the previous studies. The

operation, again, is simply to multiply the ion concentrations listed below by the ratio of hardness

values presented below and the earlier test waters.

Note that additional data were not available for Blaine Creek, KY, or Pinto Creek, AZ, and

although additional data were obtained from the City of Sacramento, CA, regarding the American

River, the default DOC value (8.2 mg/L) for California streams may be artificially high on the basis

of reported values of DOC in the Sacramento River (1.2 mg C/L), of which the American River is a

tributary. Therefore, the data from Finlayson and Verrue (1982) have been relegated to “ other data.”

Likewise, Amy Bayou is a highly contaminated and dynamic system (Don Cherry, personal

communication), and BLM normalization is not recommended for these data. A large annual

variability in water quality also excludes the use of surrogate STORET data for the Embarrass River,

MN, for BLM analysis (Lind et al. manuscript).

American River, CA (Appendix C-9). Source: Ron Myers, City of Sacramento, CA, Water Quality Laboratory

Applied to:

Hardness

(mg/L

CaCO3)

Alkalinity

(mg/L

CaCO3) pH DOC

Specific Ions (mg/L)

Ca Mg Ca:Mg Na K Cl SO4

Finlayson and

Verrue 1982

21 22 7.5 -a 5.6 1.8 2.0 3.0 - 2.6 3.8

a DOC and K data for the American River were not available.

Clinch River, VA (Appendix D-5): Source: Don Cherry, VA Poly. Inst. & State Univ., Blacksburg,VA

Applied to:

Hardness

(mg/L

CaCO3)

Alkalinity

(mg/L

CaCO3) pH DOC

Specific Ions (mg/L)

Ca Mg Ca:Mg Na K Cl SO4

Belanger et al.

1989, and

Belanger and

Cherry 1990

150 150 8.3 2.3 42 11 2.3 12 2.4 9.2 19
C-21

S. Kawishiwi River, MN (Appendix C-10). Source: STORET

Applied to:

Hardness

(mg/L

CaCO3)

Alkalinity

(mg/L

CaCO3) pH DOC

Specific Ions (mg/L)

Ca Mg Ca:Mg Na K Cl SO4

Lind et al.

manuscript

24 18 6.6 -a 5.6 2.4 1.5 1.3 0.5 1.0 4.9

a DOC data for this river were not available. TOC measurements reported by Lind et al. (manuscript) should be

adjusted based on a mean DOC:TOC ratio (0.8721) in Minnesota streams (see Section 2.3 and Appendix D-2).

Lake One, MN (Appendix C-10). Source: STORET

Applied to:

Hardness

(mg/L

CaCO3)

Alkalinity

(mg/L

CaCO3) pH DOC

Specific Ions (mg/L)

Ca Mg Ca:Mg Na K Cl SO4

Lind et al.

manuscript

10 15 6.7 -a 2.8 0.7 1.8 0.1 0.3 0.2 4.2

a DOC data for this lake were not available. TOC measurements reported by Lind et al. (manuscript) should be

adjusted based on a mean DOC:TOC ratio (0.9677) in Minnesota lakes (see Section 2.3 and Appendix D-2).

Colby Lake, MN (Appendix C-10). Source: STORET

Applied to:

Hardness

(mg/L

CaCO3)

Alkalinity

(mg/L

CaCO3) pH DOC

Specific Ions (mg/L)

Ca Mg Ca:Mg Na K Cl SO4

Lind et al.

manuscript

56 33 7.1 -a 13.3 5.4 1.6 4.0 1.4 7.3 23

a DOC data for this lake were not available. TOC measurements reported by Lind et al. (manuscript) should be

adjusted based on a mean DOC:TOC ratio (0.9677) in Minnesota lakes (see Section 2.3 and Appendix D-2).

Cloquet Lake, MN (Appendix C-10). Source: STORET

Applied to:

Hardness

(mg/L

CaCO3)

Alkalinity

(mg/L

CaCO3) pH DOC

Specific Ions (mg/L)

Ca Mg Ca:Mg Na K Cl SO4

Lind et al.

manuscript

27 21 7.2 -a 6.9 2.3 1.4 1.9b 1.4c 1.2 5.6

a DOC data for this lake were not available. TOC measurements reported by Lind et al. (manuscript) should be

adjusted based on a mean DOC:TOC ratio (0.9677) in Minnesota lakes (see Section 2.3 and Appendix D-2).

b Na data for this lake were not available. The Na value given here is based on data for Colby Lake, MN, and was

scaled on the basis of hardness (see Section 2.1): Na = 4.0 mg Na/L * (27 mg/L CaCO3 / 56 mg/L CaCO3).

c K data for this lake were not available. The K value given here is from data for Colby Lake, MN. This value was

not scaled on the basis of hardness (see discussion of K-hardness relationship in Sections 2.1 and 2.7).
D-22

Greenwood Lake (Appendix C-10), MN. Source: STORET

Applied to:

Hardness

(mg/L

CaCO3)

Alkalinity

(mg/L

CaCO3) pH DOC

Specific Ions (mg/L)

Ca Mg Ca:Mg Na K Cl SO4

Lind et al.

manuscript

17 11 6.4 -a 4 1.8 2.4 0.2b 0.3c 1.7 7.6

a DOC data for this lake were not available. TOC measurements reported by Lind et al. (manuscript) should be

adjusted based on a mean DOC:TOC ratio (0.9677) in Minnesota lakes (see Section 2.3 and Appendix D-2).

b Na data for this lake were not available. The Na value given here is based on data for Lake One, MN, and was

scaled based on hardness: Na = 0.1 mg Na/L * (17 mg/L CaCO3 / 10 mg/L CaCO3).

c K data for this lake were not available. The K value given here is from data for Lake One, MN. This value was

not scaled on the basis of hardness (see discussion of K-hardness relationship in Sections 2.1 and 2.7).

St. Louis River, MN (Appendix C-6). Source: NASQAN

Note: for the St. Louis River dataset (1973 to 1993), a question arose as to which data would be most representative

for estimating the ion concentrations in St. Louis River water for BLM analysis. In order to determine this, the

relationship between hardness and Na ion for all 20 years was plotted. Linear regression was used to fit the data.

Most data showed very high coefficient correlation (0.8-0.94). For each of these 20 regression lines, the slope and

intercept coefficients were plotted on separate graphs as functions of time (Figures 7 and 8). The following

conclusions were derived:

C A significant event occurred in 1976 and perhaps 1977 that affected the water balance of the St. Louis River. A

wastewater treatment plant was built, which substantially improved the water quality (Jesse Anderson, Minn.

Pollution Control Bd., personal communication).

C For the 1979-1993 period, hardness and ion concentrations did not change significantly as absolute values.

Therefore, general equations (which could be used to extrapolate water chemistry data till year 2000 and before

1979) can be obtained connecting hardness, alkalinity, pH, and the major ion concentrations. 

C The exponential growth in the values between 1973 and 1979 shows that averaging values on seasonal and

annual basis is not appropriate. The constant values for the slopes and intercepts for 1979-1993 allow mean

monthly and annual interpretation of the data.

C The regression equations derived for 1977 alone are recommended to predict ion concentrations based on the

water hardness levels measured in the Lind et al. (manuscript). The equations derived for each ion are

provided in Appendix D-6 with the corresponding figures.

Green-Duwamish River, WA. Source: James Buckley

Applied to:

Hardness

(mg/L

CaCO3)

Alkalinity

(mg/L

CaCO3) pH DOC

Specific Ions (mg/L)

Ca Mg Ca:Mg Na K Cl SO4

Buckley 1983 33 29 7.2 3.2a 8.9 2.8 2.0 7.5 1.2 7.0 6.3

a Value given as TOC. DOC data for this river were not available. TOC measurements reported by Buckley et al.

(1983) should be adjusted on the basis of a mean DOC:TOC ratio (0.7803) in Washington streams (see Section 2.3

and Appendix C-2).
C-23

Naugatuck River, WA. Source: STORET

Applied to:

Hardness

(mg/L

CaCO3)

Alkalinity

(mg/L

CaCO3) pH DOC

Specific Ions (mg/L)

Ca Mg Ca:Mg Na K Cl SO4

Carlson et al.

1986

39 20 6.4 3.7a 9.9 3.3 1.9 9.9 2.3 - 22

a Value given as TOC. DOC data for this river were not available. TOC measurements reported by Carlson et al.

(1986) should be adjusted on the basis of a mean DOC:TOC ratio (0.8711) in Connecticut streams (see Section 2.3

and Appendix C-2).
Total Hardness (mg/L as CaCO3)

Figure 1. Relationship between Ca and hardness in WFTS well water

C-24
Figure 2. Relationship between Mg and hardness in WFTS well water. 

Total Hardness (mg/L as CaCO3) 

C-25
Figure 3. Relationship between Na and hardness in WFTS well water. 

Total Hardness (mg/L as CaCO3)

C-26
Figure 4. Relationship between K and hardness in WFTS well water

Total Hardness (mg/L as CaCO3)

C-27
Figure 5. Relationship between Cl and hardness in WFTS well water. 

Cl (mg/L) 

Total Hardness (mg/L as CaCO3) 

C-28
Figure 6. Relationship between SO4 and hardness in WFTS well water. 

SO4 (mg/L) 

Total Hardness (mg/L as CaCO3) 

C-29
C-30

Figure 7. Slopes of the regression equations derived for Na

concentration in St. Louis River, MN, water versus water

hardness from 1973 to 1993.
C-31

Figure 8. Intercepts of the regression equations derived for Na

concentration in St. Louis River, MN water versus water

hardness from 1973 to 1993.
C-32

References

Andros, J.D. and R.R. Garton. 1980. Acute lethality of copper, cadmium, and zinc to northern squawfish. Trans.

Am. Fish. Soc. 109:235.

ASTM. 2000. Standard guide for conducting acute toxicity tests on test materials with fishes, macroinvertebrates,

and amphibians. E729-96. In: Annual book of ASTM standards. Section 11, Volume 5. American Society for

Testing and Materials, Pennsylvania, PA.

Belanger, S.E. and D.S. Cherry. 1990. Interacting effects of pH acclimation, pH, and heavy metals of acute and

chronic toxicity to Ceriodaphnia dubia (Cladoceran). J. Crustacean Biol. 10(2):225-235.

Belanger, S.E., J.L. Farris and D.S. Cherry. 1989. Effects of diet, water hardness, and population source on acute

and chronic copper toxicity to Ceriodaphnia dubia. Arch. Environ. Contam. Toxicol. 18(4):601-611.

Bennett, W.A., A. Sosa and T.L. Beitinger. 1995. Oxygen tolerance of fathead minnows previously exposed to

copper. Bull. Environ. Contam. Toxicol. 55(4):517-524.

Berthouex, P.M. and L.C. Brown. 1994. Statistics for environmental engineers. Lewis Publishers/CRC Press,

Boca Raton, FL.

Biesinger, K.E. and G.M. Christensen. 1972. Effects of various metals on survival, growth, reproduction, and

metabolism of Daphnia magna. Jour. Fish Res. Board Can. 29:1691.

Brungs, W.A., E.N. Leonard and J.M. McKim. 1973. Acute and long-term accumulation of copper by the brown

bullhead, Ictalurus nebulosus. J. Fish. Res. Board Can. 30(4):583-586.

Buckley, J.A. 1983. Complexation of copper in the effluent of a sewage treatment plant and an estimate of its

influence on toxicity to coho salmon. Water Res. 17(12):1929-1934.

Cairns, J., Jr., K.W. Thompson and A.C. Hendricks. 1981. Effects of fluctuating, sublethal applications of heavy

metal solutions upon the gill ventilation response of bluegills (Lepomis macrochirus). EPA-600/3-81-003. National

Technical Information Service, Springfield, VA.

Carlson, A.R., H. Nelson and D. Hammermeister. 1986. Development and validation of site-specific water quality

criteria for copper. Environ. Toxicol. Chem. 5(11):997-1012.

Chakoumakos, C., R.C. Russo and R.V. Thurston. 1979. The toxicity of copper to cutthroat trout (Salmo clarki)

under different conditions of alkalinity, pH, and hardness. Environ. Sci. Technol. 13:213-219.

Chapman, G.A. 1975. Toxicity of copper, cadmium and zinc to Pacific Northwest salmonids. U.S. EPA, Corvallis,

OR.

Chapman, G.A. 1978. Toxicities of cadmium, copper, and zinc to four juvenile stages of chinook salmon and

steelhead. Trans. Am. Fish. Soc. 107:841-847.

Chapman, G.A. and D.G. Stevens. 1978. Acute lethal levels of cadmium, copper, and zinc to adult male coho

salmon and steelhead. Trans. Am. Fish. Soc. 107:837-840.

Collyard, S.A., G.T. Ankley, R.A. Hoke and T. Goldstein. 1994. Influence of age on the relative sensitivity of

Hyalella azteca to diazinon, alkylphenol ethoxylates, copper, cadmium and zinc. Arch. Environ. Contam. Toxicol.

26(1):110-113.

Di Toro, D.M., H.E. Allen, H.L. Bergman, J.S. Meyer, P.R. Paquin and R.C. Santore, 2001. A Biotic Ligand

Model of the Acute Toxicity of Metals. I. Technical Basis, Environmental Toxicology and Chemistry. 20:2383-

2396.

Di Toro, D.M., H. Allen, H. Bergman, J. Meyer, R.C. Santore and P.R. Paquin. 2000. The Biotic Ligand Model:

A computational approach for assessing the ecological effects of copper and other metals in aquatic systems.

International Copper Association, Ltd.

Dixon, D.G. and J.B. Sprague. 1981a. Acclimation to copper by rainbow trout (Salmo gairdneri): A modifying

factor in toxicity. Can. J. Fish. Aquat. Sci. 38:880-888.
C-33

Dolan, D.M. and A.H. El-Shaarawi. 1991. Application of mass balance approach with censored data. J. Gr. Lakes

Res. 17:220-228.

Erickson, R.J. 1985. Analysis of major ionic content of selected U.S. waters and application to experimental design

for the evaluation of the effect of water chemistry on the toxicity of copper. Draft report. U.S. EPA, Duluth, MN.

Erickson, R.J., D.A. Benoit, V.R. Mattson, H.P. Nelson, Jr. and E.N. Leonard. 1996b. The effects of water

chemistry on the toxicity of copper to fathead minnows. Environ. Toxicol. Chem. 15(2):181-193.

Faust, S.D. and O.M. Aly. 1981. Chemistry of natural waters. Ann Arbor Science Publishers, Inc., Ann Arbor, MI.

Finlayson, B.J. and K.M. Verrue. 1982. Toxicities of copper, zinc, and cadmium mixtures to juvenile chinook

salmon. Trans. Am. Fish. Soc. 111:645-650.

Geckler, J.R., W.B. Horning, T.M. Neiheisel, Q.H. Pickering, E.L. Robinson and C.E. Stephan. 1976. Validity of

laboratory tests for predicting copper toxicity in streams. EPA-600/3-76-116. National Technical Information

Service, Springfield, VA.

Hartwell, S.I., J.H. Jin, D.S. Cherry and J. Cairns, Jr. 1989. Toxicity versus avoidance response of golden shiner,

Notemigonus crysoleucas, to five metals. J. Fish Biol. 35(3):447-456.

Horning, W.B. and T.W. Neiheisel. 1979. Chronic effect of copper on the bluntnose minnow, Pimephales notatus

(Rafinesque). Arch. Environ. Contam. Toxicol. 8:545-552.

Howarth, R.S. and J.B. Sprague. 1978. Copper lethality to rainbow trout in waters of various hardness and pH.

Water Res. 12:455-462.

Lind, D., Alto and Chatterton. Manuscript. Regional copper-nickel study: Aquatic toxicology study. Preliminary

draft report, Minnesota Environmental Quality Board, St. Paul, MN.

Lorz, H.W. and B.P. McPherson. 1976. Effects of copper or zinc in fresh water on the adaptation to sea water and

ATPase activity, and the effects of copper on migratory disposition of coho salmon (Oncorhynchus kisutch). J. Fish.

Res. Board Can. 33:2023.

McCrady, J.K. and G.A. Chapman. 1979. Determination of copper completing capacity of natural river water, well

water and artificially reconstituted water. Water Res. 13:143-150.

McGeer, J.C., R.C. Playle, C.M. Wood and F. Galvez. 2000. A physiologically based biotic ligand model for

predicting the acute toxicity of waterborne silver to rainbow trout in freshwaters. Environ. Sci. Technol. 34:4199-

4207.

Mount, D.I. 1968. Chronic toxicity of copper to fathead minnows (Pimephales promelas Rafinesque). Water Res.

2:215-223.

Mount, D.I. and C.E. Stephan. 1969. Chronic toxicity of copper to the fathead minnow (Pimephales promelas) in

soft water. J. Fish. Res. Board Can. 26:2449-2457.

Mudge, J.E., T.E. Northstrom, G.S. Jeane, W. Davis and J.L. Hickam. 1993. Effect of varying environmental

conditions on the toxicity of copper to salmon. In: Environmental toxicology and risk assessment. Gorsuch, J.W.,

F.J. Dwyer, C.G. Ingersoll and T.W. LaPoint (Eds.). ASTM STP 1216. American Society for Testing and

Materials, Philadelphia, PA. pp. 19-33.

Nebeker, A.V., M.A. Cairns and C.M. Wise. 1984a. Relative sensitivity of Chironomus tentans life stages to

copper. Environ. Toxicol. Chem. 3(1):151-158. 

Nebeker, A.V., M.A. Cairns, S.T. Onjukka and R.H. Titus. 1986a. Effect of age on sensitivity of Daphnia magna

to cadmium, copper and cyanazine. Environ. Toxicol. Chem. 5(6):527-30.

Nebeker, A.V., A. Stinchfield, C. Savonen and G.A. Chapman. 1986b. Effects of copper, nickel and zinc on three

species of Oregon freshwater snails. Environ. Toxicol. Chem. 5(9):807-811.

Pickering, Q.H., W. Brungs and M. Gast. 1977. Effect of exposure time and copper concentration on reproduction of

the fathead minnow (Pimephales promelas). Water Res. 11:1079-1083. 
C-34

Rice, D.W., Jr. and F.L. Harrison. 1983. The sensitivity of adult, embryonic, and larval crayfish Procambaris

clarkii to copper. UCRL-53048. National Technical Information Service, Springfield, VA.

Richards, V.L. and T.L. Beitinger. 1995. Reciprocal influences of temperature and copper on survival of fathead

minnows, Pimephales promelas. Bull. Environ. Contam. Toxicol. 55(2):230-6.

Samuelson, D.F. 1976. Water quality: Western Fish Toxicology Station and Western Oregon rivers. EPA-600/3-

76-077. U.S. EPA, Duluth, MN.

Schubauer-Berigan, M.K., J.R. Dierkes, P.D. Monson and G.T. Ankley. 1993. pH-dependent toxicity of cadmium,

copper, nickel, lead and zinc to Ceriodaphnia dubia, Pimephales promelas, Hyalella azteca and Lumbriculus

variegatus. Environ. Toxciol. Chem. 12(7):1261-1266.

Seim, W.K., L.R. Curtis, S.W. Glenn and G.A. Chapman. 1984. Growth and survival of developing steelhead

trout (Salmo gairdneri) continuously or intermittently exposed to copper. Can. J. Fish. Aquat. Sci. 41(3):433-438.

Spehar, R.L. and J.T. Fiandt. 1986. Acute and chronic effects of water quality criteria-based metal mixtures on three

aquatic species. Environ. Toxicol. Chem. 5(10):917-931.

Stephan, C.E., D.I. Mount, D.J. Hansen, J.H. Gentile, G.A. Chapman and W.A. Brungs. 1985. Guidelines for

deriving numerical national water quality criteria for the protection of aquatic organisms and their uses. PB85-

227049. National Technical Information Service, Springfield, VA.

Thompson, K.W., A.C. Hendricks and J. Cairns, Jr. 1980. Acute toxicity of zinc and copper singly and in

combination to the bluegill (Lepomis macrochirus). Bull. Environ. Contam. Toxicol. 25:122.

U.S. EPA. 1993. Methods for measuring the acute toxicity of effluents and receiving water to freshwater and marine

organisms. Fourth Edition. EPA/600/4-90/027F. National Technical Information Service, Springfield, VA.

U.S. EPA. 1998a. Ambient water quality criteria derivation methodology human health technical support

document. EPA-822-B-98-005. Office of Water. Washington, DC.

U.S. EPA. 1998b. Ambient aquatic life water quality criteria for copper. Draft Report. Office of Water. Washington,

DC.

U.S. EPA. 2001. 2001 update of ambient water quality criteria for cadmium. EPA-822-R-01-001. Office of Water.

Washington, DC.

Winner, R.W. 1985. Bioaccumulation and toxicity of copper as affected by interactions between humic acid and

water hardness. Water Res. 19(4):449-455.

WPPSS (Washington Public Power Supply System). 1982. Supply system nuclear project No. 3. Environmental

Report - Operating License Stage Docket No. 50-508. Richland, WA.
C-35

Appendix C-1. Calculations for Ionic Composition of Standard 

Laboratory-Reconstituted Water

Molecular Weights Atomic Weights

NaHCO3 = 84.03

CaSO4

.2H2

 O = 172.12

MgSO4= 120.37

KCl = 74.55

SO4

 = 96.06

Na = 22.98

Ca = 40.08

Mg = 24.31

K = 39.10

Cl = 35.45

Example Calculation

[Na] in very soft water:

12 mg NaHCO3/L x 1 mmol NaHCO3/84.03 mg NaHCO3 = 0.143 mmol NaHCO3/L. 

0.143 mmol NaHCO3/L x (1 mmol Na/1 mmol NaHCO3) x 22.98 mg Na/1 mmol Na = 3.3 mg Na/L.

[Ca] in very soft water:

7.5 mg CaSO4

.2H2O/L x 1 mmol CaSO4

.2H2O/172.12 mg CaSO4

.2H2O = 0.044 mmol CaSO4

.2H2O/L. 

0.044 mmol CaSO4

.2H2O/L x (1 mmol Ca/1 mmol CaSO4

.2H2O) x 40.08 mg Ca/1 mmol Ca = 1.8 mg Ca/L.

[Mg] in very soft water:

7.5 mg MgSO4/L x 1 mmol MgSO4/120.37 mg MgSO4 = 0.062 mmol MgSO4/L. 

0.062 mmol MgSO4

/L x (1 mmol Mg/1 mmol MgSO4

) x 24.31 mg Mg/1 mmol Mg = 1.5 mg Mg/L.

[K] in very soft water:

0.5 mg KCl/L x 1 mmol KCl/74.55 mg KCl = 0.0067 mmol KCl/L. 

0.0067 mmol KCl/L x (1 mmol K/1 mmolKCl) x 39.102 mg K/1 mmol K = 0.26 mg K/L.

[Cl] in very soft water:

0.5 mg KCl/L x 1 mmol KCl/74.55 mg KCl = 0.0067 mmol KCl/L. 

0.0067 mmol KCl/L x (1 mmol Cl/1 mmolKCl) x 35.453 mg Cl/1 mmol K = 0.24 mg Cl/L.

[SO4] in very soft water:

7.5 mg CaSO4.2H2O/L x 1 mmol CaSO4.2H2 O/172.12 mg CaSO4.2H2O = 0.044 mmol CaSO4.2H2O/L. 

0.044 mmol CaSO4

.2H2O/L x (1 mmol SO4

/1 mmol CaSO4

.2H2O) x 96.064 mg Ca/1 mmol Ca = 4.2 mg Ca/L.

[SO4] in very soft water:

7.5 mg MgSO4

/L x 1 mmol MgSO4

/120.37 mg MgSO4

 = 0.062 mmol MgSO4

/L. 

0.062 mmol MgSO4/L x (1 mmol SO4/1 mmol MgSO4) x 96.064 mg Mg/1 mmol Mg = 6.0 mg Mg/L.

Total SO4

 = 10.2 mg/L

Conversion Factors to calculate water hardness (as CaCO3) from [Ca] and [Mg]:

[Ca] x 2.497

[Mg] x 4.116
C-36

Appendix C-2. Dissolved, Particulate, and Estimated Total Organic Carbon for Streams

and Lakes by State (as presented in EPA Document #822-B-98-005) 

Streams Lakes

State POC DOC Est. TOC Est. DOC:TOC POC DOC Est. TOC Est. DOC:TOC

AK 0.54 4.6 5.14 89.49 0.53 6.4 6.93 92.35

AL 0.72 3.4 4.12 82.52 --- --- --- ---

AR 0.8 7.2 8 90.00 0.4 2.7 3.1 87.10

AZ 0.71 5.2 5.91 87.99 0.52 4.2 4.72 88.98

CA 1.13 8.2 9.33 87.89 0.32 2.3 2.62 87.79

CO 1.29 8.6 9.89 86.96 --- --- --- ---

CT 0.71 4.8 5.51 87.11 --- --- --- ---

DC --- --- --- --- --- --- --- ---

DE* 0.7 7.1 7.8 91.03 --- --- --- ---

FL^ 0.68 16.1 16.78 95.95 2.9 12.1 15 80.67

GA 0.67 4.3 4.97 86.52 --- --- --- ---

HI 0.59 4 4.59 87.15 --- --- --- ---

IA 1.79 11.6 13.39 86.63 --- --- --- ---

ID 0.6 3.2 3.8 84.21 --- --- --- ---

IL 1.77 6.8 8.57 79.35 0.12 4.7 4.82 97.51

IN 0.71 9.2 9.91 92.84 --- --- --- ---

KS 1.75 5.2 6.95 74.82 1.53 4.5 6.03 74.63

KY 0.75 3.1 3.85 80.52 --- --- --- ---

LA 1.52 6.9 8.42 81.95 0.65 5.6 6.25 89.60

MA 0.47 5.9 6.37 92.62 --- --- --- ---

MD 1.66 3.7 5.36 69.03 --- --- --- ---

ME 0.46 15.3 15.76 97.08 --- --- --- ---

MI 0.58 6.3 6.88 91.57 0.32 2.7 3.02 89.40

MN 1.79 12.2 13.99 87.21 0.16 4.8 4.96 96.77

MO 0.56 4.2 4.76 88.24 --- --- --- ---

MT 0.9 9.4 10.3 91.26 0.91 8.2 9.11 90.01

NC 1.14 11.5 12.64 90.98 --- --- --- ---

ND 1.14 14.5 15.64 92.71 0.8 14.9 15.7 94.90

NE 1.84 6.8 8.64 78.70 --- --- --- ---

NH 0.28 4.2 4.48 93.75 --- --- --- ---

NJ 0.69 5.5 6.19 88.85 1.04 5 6.04 82.78

NM 1.43 6.3 7.73 81.50 0.51 5.2 5.71 91.07

NV 0.82 4.2 5.02 83.67 --- --- --- ---

NY 1.4 4 5.4 74.07 0.46 2.4 2.86 83.92

OH 0.57 5 5.57 89.77 0.49 2.6 3.09 84.14

OK^ 1.27 7.7 8.97 85.84 1.72 15 16.72 89.71

OR*^ 1.14 2.1 3.24 64.81 0.64 4.4 5.04 87.30

PA 2.19 5.4 7.59 71.15 0.63 3.2 3.83 83.55

RI* 0.42 8.3 8.72 95.18 --- --- --- ---

SC 0.7 5.7 6.4 89.06 --- --- --- ---

SD 1.25 7.6 8.85 85.88 --- --- --- ---

TN 0.67 2.3 2.97 77.44 --- --- --- ---

TX 1.33 6.5 7.83 83.01 1.55 10.3 11.85 86.92

UT^ 1.38 8.9 10.28 86.58 0.5 2.4 2.9 82.76

VA 0.81 4.7 5.51 85.30 --- --- --- ---

VT 0.31 4.5 4.81 93.56 --- --- --- ---

WA 1.52 5.4 6.92 78.03 0.61 2.8 3.41 82.11

WI 1.03 9.2 10.23 89.93 0.16 4.1 4.26 96.24

WV 0.63 2.8 3.43 81.63 --- --- --- ---

WY 1.07 8.2 9.27 88.46 --- --- --- ---
Streams Lakes

State POC DOC Est. TOC Est. DOC:TOC POC DOC Est. TOC Est. DOC:TOC

C-37

Mean 85.71 Mean 87.84

Max 97.08 Max 97.51

Min 64.81 Min 74.63

* States where sample size was low for streams.

^ States where sample size was low for lakes.
C-38

Appendix C-3. Mean TOC and DOC in Lake Superior Dilution Water 

 (data from Greg Lien, U.S. EPA-Duluth, MN)

Replicate Ambient (8/29/2000) pH 7.0 (8/30/2000) pH 6.2 (8/31/2000)

Filter Blank* -0.04 0.22 0.38

Pre-gill 

experiment TOC

a 1.13 1.34 1.26

b 1.37 1.30 1.36

Mean 1.25 1.32 1.31

Post-gill

experiment TOC

a 1.20 1.24 1.18

b 1.27 1.46 1.10

Mean 1.24 1.35 1.14

Pre-gill

experiment DOC

a 1.96 1.51 1.34

b 1.52 1.28 0.99

Mean 1.74 1.40 1.17

Post-gill

experiment DOC

a 1.49 1.36 1.44

b 1.64 1.58 1.24

Mean 1.57 1.47 1.34

* Filter blank is ultra-pure Duluth-EPA laboratory water.
C-39

Appendix C-4. Measured Hardness and Major Ion and Cation Concentrations 

in WFTS Well Water from April 1972 to April 1978. Concentrations Given as Mg/L 

(data from Samuelson 1976 and Chapman, personal communication)

Month Total Hardness Ca Mg Na K SO4 Cl

Mar-72

Apr-72 7.9 2 5 1.1 <10.0 8

May-72 22 5.8 1.4 4.4 0.5 <5.0 7

Jun-72 24 5.8 1.6 4.4 0.5 3 7

Jul-72 23 6.7 1.6 4.6 0.5 <1.0 8.3

Aug-72 23 6.5 1.7 4.7 0.5 <10.0 6.3

Sep-72 22 6 1.6 4.5 0.6 <10.0 4

Oct-72 22 6.7 1.9 4.7 0.6 5 5.5

Nov-72 23 6.2 1.6 4.2 0.6 3.7 5.3

Dec-72 23 6.2 1.5 4.2 0.5 3 4

Jan-73 52 15.3 3.5 7.1 0.7 7.8 12.4

Feb-73 33 7.7 2.1 5 0.5 5 5

Mar-73 30 8 2.1 5.3 0.7 5 6

Apr-73 31 8.9 2.3 5.4 0.7 5.3 8.8

May-73 28 8.3 2.4 5.8 0.7 3 8

Jun-73 28 8.4 2.2 5.8 0.7 4.8 7.5

Jul-73 26 7.4 1.9 5.8 0.8 <5.0 6.8

Aug-73 25 6.5 1.7 5.7 0.7 3.1 5.8

Sep-73 25 6.7 1.7 5.4 0.7 3.1 5.3

Oct-73 27 7 1.8 5.4 0.7 2.9 5.4

Nov-73 28 7.9 2.1 4.8 0.7 10 6.8

Dec-73 62 20.3 4.2 9 0.8 13 14

Jan-74 67 21.3 4.8 7 0.8 17.3 11.3

Feb-74 58 14.3 3.4 6.9 0.9 14.7 6.7

Mar-74 53 20.8 3.8 7.2 0.7 13 7

Apr-74 51 18.2 3.7 6.8 0.6 15.5 8.5

May-74 23 7.5 2.1 4.6 0.6 5 4.8

Jun-74 22 6 1.9 4.8 0.5 3 4.5

Jul-74 23 5.4 1.7 5 0.6 3.3 6.3

Aug-74 23 4.8 1.6 5 0.7 3 6

Sep-74 23 5.8 1.5 5.1 0.7 2.9 4.8

Oct-74 23 11 2 7.1 0.8 3.1 5

Nov-74 23 12 2.6 4.5 0.5 3.8 5.3

Dec-74 24 6.4 2.5 5.2 0.7 3.8 5

Jan-75 41 7.7 2.9 6.7 0.6 8 8

Feb-75 61 11.6 4.2 8.6 0.8 16 11.8

Mar-75 54 9.1 3.1 6.4 0.6 8 8

Apr-75 4.4 1.6 4.4 0.5 3 5

May-75 7.2 2 5 0.5 6 7

Jun-75 4.4 1.6 4.6 0.6 5 6

Jul-75 5.2 1.6 7 0.7 5 7

Aug-75 5.2 1.4 7 0.6 5 5

Sep-75 4.5 1.5 4.5 0.7 5 4

Oct-75 7.1 1.9 4.3 0.5 20 5

Nov-75 18 5.3 1.5 4.2 0.5 5 4

Dec-75

Jan-76

Feb-76 9.8 5 5.4 0.4 9 9

Mar-76 4.1 0.1 3 6

Apr-76 5.3 0.1 6 9
Month Total Hardness Ca Mg Na K SO4 ClD

C-40

May-76 7.9 1.8 4.5 0.5 3 6

Jun-76 27 8.1 1.9 3.3 0.6 4 7

Jul-76 26

Aug-76 23 4.9 1.3 4.8 0.1 3 6

Sep-76 23 6.7 2.6 4.7 0.1

Oct-76 21 6.7 2.6 4.7 0.1

Nov-76 22 7.7 3 4.7 0.1 3

Dec-76 25.5 6.4 1.8 5 0.1 4 7

Jan-77 27.2 7.7 2.6 5.6 0.6 4 8

Feb-77 10.7 4.9 5.9 0.6 3 11

Mar-77 3 8

Apr-77 10.7 2.2 5.5 0.8 3 7

May-77 25 5 1.8 5 0.8 3 5

Jun-77 27 6.6 2 5.2 0.7 3 5

Jul-77 24 6.7 2 7.1 0.8 3 7

Aug-77 25 6.9 1.9 6.9 1 8

Sep-77 27 9.9 2.1 5.9 0.9 3 6

Oct-77 3

Nov-77 6.6 2.1 5.6 0.9 10 4.6

Dec-77 27 9.7 4.95 0.65 9 4.6

Jan-78 10.9 3.75 0.85 6 12

Feb-78 10.6 3.8 8.6 0.7 5 11

Mar-78 10.2 2.6 4.7 0.6 6 9

Apr-78 8.3 2.4 0.7 5 9.55
Date pH Hardness Alkalinity Ca Mg Na K Cl SO4 NO3 DOCD

C-41

19790329 7.6 80 63 19 8 8.4 2.3 7.8 13

19790430 7.6 37 29 8.7 3.7 2.2 1.3 2.8 8.9 20

19790611 7.2 47 34 11 4.8 3.1 0.8 2.8 9.4

19790723 7.6 73 55 17 7.3 3.9 0.9 3.7 8.9 30

19790827 7.2

19791015 8.1 74 54 16 8.2 5 1.1 3.9 13 0.01 12

19791126 7.8 61 52 14 6.3 3.8 0.9 3.6 11 0.37

19800121 7.6 60 53 14 6 3.8 0.9 3.2 9.9 0.15

19800219 7.4 63 51 15 6.2 3.9 0.8 2.9 9.2 0.19 17

19800331 8.4 68 64 16 6.9 4.2 1.1 3.5 9.2 0.3

19800602 8.3 84 72 19 8.8 6.4 1.2 5 15 0.01 21

19800630 8.3 93 68 21 9.9 7.9 1.4 6.7 24 0.02

19800804 8.1 130 110 28 14 10 1.9 11 24 0.01 13

19800902 7.8 110 82 24 11 7.2 1.7 7.6 18 0.01

19800929 7.6 73 54 16 8.1 5.7 1.4 5.8 14 0.12

19801103 7 82 58 18 8.9 5.6 1.3 6.9 18 0.19 23

19801208 67 50 15 7.2 4.6 1 4.1 11 0.19

19810105 7.6 70 55 16 7.2 4.2 1.1 4.1 13 0.23

19810209 7.5 68 58 16 6.9 4.9 1 3.5 8.1 0.27 14

19810309 7.7 61 57 14 6.2 5.2 1.8 5.1 8.6 0.36

19810504 7.3 42 40 9.6 4.3 3.7 1.2 3.6 9.6 0.18 21

19810706 7.4 51 39 12 5 3.5 1.2 3.2 7.5 0.14 10

19810908 7.9 73 64 16 8 4.2 0.8 4.2 8.3 0.11

19811020 7.6 51 37 12 5.2 4.3 1.2 4.2 8.9 0.31

19820113 62 52 14 6.5 4 0.9 3.7 9.3 0.24

19820309 7.4 66 58 15 7 5.3 1 3.8 11 0.36

19820420 7.2 32 25 7.5 3.3 2.1 1.3 2.3 6 0.19

19820621 7.9 61 55 14 6.4 4.3 1.1 4 10 0.1

19820809 7.4 66 54 15 6.9 3.9 0.6 3.5 9 0.25

19821004 8 73 63 15 8.7 4.9 1 4.7 13 0.11

19821207 7.3 55 43 12 6.1 4.2 0.8 3.3 16 0.24

19830131 6.9 62 50 14 6.5 4.1 0.8 3.5 15 0.36

19830328 7.5 68 56 15 7.3 4.5 1.2 4.1 15 0.35

19830523 8.2 68 53 15 7.5 4 1.3 0.8 23 0.12

19830718 7.6 67 53 15 7.2 3.7 1.3 3.7 22 0.15

19831031 7.7 64 48 14 7 3.9 1.2 3.5 24 0.12

19840109 7.4 57 50 13 6 3.6 0.9 3.4 13 0.23

19840306 7.1 66 57 15 7 4.4 0.9 5.2 8.7 0.31

19840424 7.2 51 39 11 5.6 3.1 1.4 3.2 14 0.12

19840619 9.5 52 39 12 5.3 2.9 0.8 3.6 10 0.13

19840822 6.4 70 58 15 7.9 4.7 1 3.8 17 0.1

19841009 7.6 73 16 7.9 4.6 1 3.7 15 0.1

19841120 7.1 64 14 7.1 3.9 0.9 3.7 14 0.24

19850211 7 69 15 7.7 4.6 1.1 4 11 0.27

19850325 7.3 61 13 7 5.6 2.5 6.6 16 0.31

19850506 7.4 55 12 6 3.6 1.7 4.2 14 0.15

19850730 7.6 62 14 6.6 3.2 0.9 4 9.8 0.1

19851021 7.5 58 12 6.8 3.7 1.1 0.2 12 0.13
C-42

Appendix C-6. Water Composition of St. Louis River, MN, from USGS NASQAN and 

Select Relationships to Water Hardness

Date pH Hardness Alkalinity Ca Mg Na K Cl SO4 NO3 DOC

19730222 6.8 68 53 17 6.3 11 1.6 14 14 0.19

19730503 7.1 58 46 14 5.5 6.6 1.1 9.5 13 0.17

19730816 6.9 70 51 17 6.6 7.6 1.2 9 20 0.01

19731128 7 65 48 16 6.1 7.5 1.3 8.8 14

19740221 7 64 48 16 5.8 8.9 1.3 12 14

19740516 6.9 45 32 11 4.3 3.5 1.2 3.8 11

19740919 88 60 21 8.6 12 1.8 17 23

19741030 7.3 83 62 23 6.3 13 1.3 16 23

19741209 7.4 86 62 22 7.6 12 1.6 15 18

19750121 7.3 74 66 18 7 10 1.1 12 13

19750303 7.3 74 68 17 7.6 10 1.7 11 12

19750407 7.2 95 80 22 9.7 11 2 14 16

19750527 7.5 63 50 15 6.1 8.5 1.5 9.2 12

19750708 9.2 58 43 14 5.7 3.2 1 3.4 10

19750818 7.2 73 56 18 6.9 12 1.3 16 16

19750929 7.4 90 72 23 8 12 1.5 13 20

19751110 7.1 90 63 22 8.4 12 1.7 15 24

19751216 7.6 87 61 22 7.8 14 1.6 16 28

19760209 7.5 72 59 18 6.6 13 1.6 13 18

19760322 7.7 78 65 19 7.4 12 1.4 11 17

19760503 7.6 59 43 14 5.8 7.9 1.3 8.6 15

19760614 7.5 94 75 22 9.4 16 1.9 20 20

19760726 7.4 93 80 22 9.3 21 1.9 25 24

19760908 7.5 82 78 18 9.1 17 2.5 9.3 26

19761019 7.5 83 72 20 8.1 21 1.6 24 21

19761129 7.4 95 74 22 9.7 25 1.8 32 24

19770110 7.3 85 88 20 8.4 17 1.5 15 19

19770214 8.2 82 73 20 7.8 18 1.7 26 17

19770404 7.3 87 67 21 8.5 20 2.4 28 24

19770516 7.3 120 98 29 11 30 2.8 26 36

19770628 7.8 100 75 24 9.9 13 2 16 23

19770808 7.4 110 90 26 10 27 2.2 32 28

19770919 7.4 73 44 17 7.3 6.6 1.7 8.9 17

19771031 7.6 64 47 15 6.5 7.9 1.3 9.7 22 37

19771212 7.5 65 50 15 6.8 6.3 1.2 7.1 16

19780123 7.3 71 52 17 6.9 12 1.5 9.4 18

19780306 7.2 67 48 16 6.5 8.8 1.2 17 16 32

19780417 7.5 43 28 10 4.3 4.2 1.8 5.7 15

19780530 7.9 64 54 15 6.4 5.7 1.5 7.1 14 33

19780710 7.4 53 44 13 5.1 4.3 1.3 5.3 8.9

19780821 8.4 60 42 15 5.5 5.3 1.5 6.5 12 36

19781002 7.7 71 57 17 6.9 8.2 1.1 9.6 15 24

19781115 7.4 68 52 16 6.8 11 1.1 10 12

19781218 7.4 68 55 16 6.9 11 1 9.2 14

19790205 7.4 63 57 15 6.3 334.4 1 3.1 8 12
Date pH Hardness Alkalinity Ca Mg Na K Cl SO4 NO3 DOCD

C-43

19790329 7.6 80 63 19 8 8.4 2.3 7.8 13

19790430 7.6 37 29 8.7 3.7 2.2 1.3 2.8 8.9 20

19790611 7.2 47 34 11 4.8 3.1 0.8 2.8 9.4

19790723 7.6 73 55 17 7.3 3.9 0.9 3.7 8.9 30

19790827 7.2

19791015 8.1 74 54 16 8.2 5 1.1 3.9 13 0.01 12

19791126 7.8 61 52 14 6.3 3.8 0.9 3.6 11 0.37

19800121 7.6 60 53 14 6 3.8 0.9 3.2 9.9 0.15

19800219 7.4 63 51 15 6.2 3.9 0.8 2.9 9.2 0.19 17

19800331 8.4 68 64 16 6.9 4.2 1.1 3.5 9.2 0.3

19800602 8.3 84 72 19 8.8 6.4 1.2 5 15 0.01 21

19800630 8.3 93 68 21 9.9 7.9 1.4 6.7 24 0.02

19800804 8.1 130 110 28 14 10 1.9 11 24 0.01 13

19800902 7.8 110 82 24 11 7.2 1.7 7.6 18 0.01

19800929 7.6 73 54 16 8.1 5.7 1.4 5.8 14 0.12

19801103 7 82 58 18 8.9 5.6 1.3 6.9 18 0.19 23

19801208 67 50 15 7.2 4.6 1 4.1 11 0.19

19810105 7.6 70 55 16 7.2 4.2 1.1 4.1 13 0.23

19810209 7.5 68 58 16 6.9 4.9 1 3.5 8.1 0.27 14

19810309 7.7 61 57 14 6.2 5.2 1.8 5.1 8.6 0.36

19810504 7.3 42 40 9.6 4.3 3.7 1.2 3.6 9.6 0.18 21

19810706 7.4 51 39 12 5 3.5 1.2 3.2 7.5 0.14 10

19810908 7.9 73 64 16 8 4.2 0.8 4.2 8.3 0.11

19811020 7.6 51 37 12 5.2 4.3 1.2 4.2 8.9 0.31

19820113 62 52 14 6.5 4 0.9 3.7 9.3 0.24

19820309 7.4 66 58 15 7 5.3 1 3.8 11 0.36

19820420 7.2 32 25 7.5 3.3 2.1 1.3 2.3 6 0.19

19820621 7.9 61 55 14 6.4 4.3 1.1 4 10 0.1

19820809 7.4 66 54 15 6.9 3.9 0.6 3.5 9 0.25

19821004 8 73 63 15 8.7 4.9 1 4.7 13 0.11

19821207 7.3 55 43 12 6.1 4.2 0.8 3.3 16 0.24

19830131 6.9 62 50 14 6.5 4.1 0.8 3.5 15 0.36

19830328 7.5 68 56 15 7.3 4.5 1.2 4.1 15 0.35

19830523 8.2 68 53 15 7.5 4 1.3 0.8 23 0.12

19830718 7.6 67 53 15 7.2 3.7 1.3 3.7 22 0.15

19831031 7.7 64 48 14 7 3.9 1.2 3.5 24 0.12

19840109 7.4 57 50 13 6 3.6 0.9 3.4 13 0.23

19840306 7.1 66 57 15 7 4.4 0.9 5.2 8.7 0.31

19840424 7.2 51 39 11 5.6 3.1 1.4 3.2 14 0.12

19840619 9.5 52 39 12 5.3 2.9 0.8 3.6 10 0.13

19840822 6.4 70 58 15 7.9 4.7 1 3.8 17 0.1

19841009 7.6 73 16 7.9 4.6 1 3.7 15 0.1

19841120 7.1 64 14 7.1 3.9 0.9 3.7 14 0.24

19850211 7 69 15 7.7 4.6 1.1 4 11 0.27

19850325 7.3 61 13 7 5.6 2.5 6.6 16 0.31

19850506 7.4 55 12 6 3.6 1.7 4.2 14 0.15

19850730 7.6 62 14 6.6 3.2 0.9 4 9.8 0.1

19851021 7.5 58 12 6.8 3.7 1.1 0.2 12 0.13
Date pH Hardness Alkalinity Ca Mg Na K Cl SO4 NO3 DOCD

C-44

19851203 7.4 73 16 8 4 1 4.2 18 0.16

19860303 7.4 66 15 7 4 1 3.4 10 0.24

19860407 7.3 0.19

19860602 7.5 58 13 6.3 3.5 1 2.8 15 0.1

19860818 7.9 74 15 8.9 4.6 1.2 3.7 24 0.1

19861112 7.5 55 12 6 3.4 1.4 3.8 19 0.27

19861210 7.3 70 57 13 9 5 1 4.8 21 0.16

19870218 7 66 15 6.8 3.7 0.9 3.1 12 0.24

19870518 8 83 18 9.3 5.8 1.2 5 10 0.1

19870622 7.8 75 16 8.5 6.2 1.1 5.2 19 0.1

19870721 7.6 51 12 5.2 2.8 1.3 3.1 15 0.1

19871028 8 82 17 9.6 6.8 1.4 1.3 19 0.1

19871208 7.9 69 15 7.7 5.3 1.4 4.8 17 0.1

19880119 7.4 73 16 8 5.1 1 3.6 15 0.15

19880223 7.4 85 19 9.2 6.5 8.5 5.1 16 0.2

19880412 7.4 42 9.2 4.7 3 2.8 5 20 0.25

19880907 7.1 70 15 8 5.3 1.5 6.1 18 0.15

19881031 7.6 100 21 12 9 1.9 7.8 27 0.1

19881130 7.6 78 17 8.6 5.5 1.3 5.5 19 0.19

19890221 7.1 77 17 8.4 6.3 1.3 4.4 17 0.25

19890410 7.2 48 11 5 4.9 1.8 8.1 8 0.37

19890626 7.4 63 14 6.8 4.6 1.1 5 12 0.15

19890814 8.1 95 20 11 9.1 1.5 8.9 18 0.1

19891101 8.1 110 20 15 7.8 1.9 6.3 31 0.1

19891218 7.5 88 17 11 6.1 1.4 5 22 0.16

19900123 7.3 100 18 14 7.2 1.7 5.2 28 0.23

19900416 7.5 62 13 7.2 5.1 1.9 5.4 14 0.2

19900716 7.7 70 15 8 5.7 1.3 5.4 11 0.2

19900820 8.1 95 20 11 7.8 1.5 7.9 20 0.1

19901009 7.3 81 18 8.7 5.4 1.5 5.7 13 0.1

19910102 7.4 83 19 8.7 5.3 1.4 5 12 0.2

19910212 7.1 80 18 8.5 6.8 1.3 3.9 11 0.2

19910502 6.7 56 13 5.8 4 1 3.7 7.9 0.1

19910610 7.3 64 15 6.5 4 0.7 4.1 6.9 0.12

19910731 7.8 55 13 5.4 2.5 1 2.6 3.8 0.05

19910801 7.3

19911003 7.8 67 15 7.1 4.4 1 4.4 9.6 0.068

19911204 7.4 61 13 6.9 4.8 1 3.5 7 0.18

19920113 7.9 67 15 7.2 4.3 1.1 3.2 9.3 0.21

19920413 7.7 30 7.8 2.5 2.5 0.3 2.4 4.8 0.16

19920722 7.6 71 16 7.5 4.8 0.9 2.1 9.6 0.11

19921026 8.2 86 18 10 5.3 1.2 5.4 14

19921216 7.6 89 19 10 6 1.2 5.6 13 0.25

19930201 7.2 83 18 9.1 7.3 1.2 7.3 12 0.28

19930426 7.7 66 15 6.8 4.1 1.2 4.9 9.5 0.092

19930722 7.5 64 15 6.5 4 0.2 3.9 7.7 0.079

19931201 7.7 80 17 9 4.8 1 4 11 0.16
Date pH Hardness Alkalinity Ca Mg Na K Cl SO4 NO3 DOCD

C-45

19940216 7.3

19940511 7.7 51 11 5.6 3.7 1.1 3.4 9.4 0.076

MIN 6.4 30 25 7.5 2.5 2.1 0.2 0.2 3.8 0.01 10

MAX 9.5 130 110 29 15 30 8.5 32 36 0.37 37

MEAN 7.52 71.11 56.94 16.16 7.46 7.09 1.37 7.39 15.04 0.17 22.19
C-46
C-47
C-48

Appendix C-7. Supplementary Data for Bennett et al. (1995)

Tank

Dose

(μg Cu/L)

Conductivity

(μmho/cm) pH

Oxygen

(mg/L)

Temp

(

oC)

Alkalinity

(as mg

CaCO3/L)

Hardness

(as mg

CaCO3/L)

0 hours 7/9/92

a 897 325 8.62 7.5 21 100 96

b 897 300 8.6 7.6 21 100 96

c 897 320 8.6 7.6 21 80 96

d 607 320 8.62 7.7 21 80 96

e 607 370 8.62 7.6 21 80 96

f 607 328 8.64 7.6 21 80 96

g 93 310 8.64 7.6 21 80 96

h 93 370 8.69 7.5 21 80 96

I 93 310 8.6 7.6 21 80 96

j 505 310 8.62 7.7 21 100 96

k 505 310 8.65 7.7 21 80 96

l 505 320 8.69 7.7 21 80 96

m 319 320 8.69 7.7 21 80 96

n 319 330 8.68 7.7 21 80 96

o 319 320 8.67 7.7 21 80 96

p 0 310 8.62 7.5 21 80 96

q 0 320 8.63 7.6 21 80 96

r 0 320 8.6 7.7 21 80 96

24 hours 7/10/92

a 897 300 7.78 8.5 21.5 60 104

b 897 305 7.64 8.4 22 80 100

c 897 305 7.68 8.5 22 90 100

d 607 300 7.7 8.4 21.5 90 100

e 607 305 7.65 8.4 21.5 80 100

f 607 305 7.75 8.4 21.5 80 100

g 93 300 7.77 9.1 22 80 100

h 93 295 7.76 9.2 21.5 80 108

I 93 295 7.76 9 21.5 85 100

j 505 300 7.73 8.8 22 90 84

k 505 300 7.71 8.8 21.5 80 100

l 505 300 7.73 8.7 21.5 80 100

m 319 300 7.74 9.1 21.5 80 100

n 319 300 7.52 8.5 22 80 100

o 319 310 7.79 8.7 22.5 80 100

p 0 305 7.79 9.1 22 80 100

q 0 305 7.7 9.1 22 80 104

r 0 300 7.71 9.1 22 80 104

48 hours 7/11/92

a 897 * * * * * *

b 897 * * * * * *

c 897 320 8.1 7.2 21.5 100 96

d 607 315 7.91 6.9 21.5 100 96

e 607 310 7.84 6.8 21.5 100 100

f 607 315 8 7 21.5 100 104

g 93 300 8.19 7.7 21.5 100 100
Tank

Dose

(μg Cu/L)

Conductivity

(μmho/cm) pH

Oxygen

(mg/L)

Temp

(

oC)

Alkalinity

(as mg

CaCO3/L)

Hardness

(as mg

CaCO3/L)D

C-49

h 93 300 8.13 7.7 21 100 100

I 93 300 8.16 7.6 21 100 104

j 505 310 8.1 7.5 21 80 100

k 505 310 8.12 7.4 21 100 100

l 505 310 8.13 7.4 21 80 100

m 319 310 8.12 7.4 21 100 100

n 319 310 7.8 6.4# 21.5 100 100

o 319 310 8.18 7.3 22 100 96

p 0 300 8.16 8 21.5 80 100

q 0 300 8.1 7.9 21.5 80 104

r 0 300 8.21 8 21.5 100 100

72 hours 7/12/92

a 897 * * * * * *

b 897 * * * * * *

c 897 * * * * * *

d 607 310 8.02 8.9 21.5 100 100

e 607 315 8.04 8.8 21.5 100 100

f 607 315 8.02 8.7 21.5 80 100

g 93 310 7.92 9.1 21.5 100 104

h 93 305 7.91 9.1 21 100 100

I 93 310 7.91 9 21 80 106

j 505 315 7.97 8.9 21.5 100 104

k 505 310 7.96 8.9 21 100 100

l 505 310 7.96 9 21 80 104

m 319 310 7.91 9 21 100 100

n 319 310 7.97 9 21 80 100

o 319 320 7.99 8.8 22 100 104

p 0 300 7.86 9.3 21.5 100 104

q 0 300 7.81 9.1 21.5 80 100

r 0 305 7.93 9.3 21.5 80 100

96 hours 7/13/92

a 897 * * * * * *

b 897 * * * * * *

c 897 * * * * * *

d 607 320 8.03 7.3 21.5 100 104

e 607 320 8.07 7.3 21.5 100 100

f 607 325 8.02 7.2 21.5 100 104

g 93 325 7.95 7.1 21.5 120 104

h 93 315 8.03 7.5 21 100 100

I 93 310 8.02 7.4 21 100 100

j 505 320 8.06 7.4 21.5 80 100

k 505 320 8.05 7.4 21 120 100

l 505 320 8.03 7.3 21 100 104

m 319 315 8.05 7.5 21 100 104

n 319 320 8.06 7.4 21 100 100

o 319 330 8.08 7.3 22 100 104
Tank

Dose

(μg Cu/L)

Conductivity

(μmho/cm) pH

Oxygen

(mg/L)

Temp

(

oC)

Alkalinity

(as mg

CaCO3/L)

Hardness

(as mg

CaCO3/L)D

D-50

p 0 330 7.78 8.1 21.5 80 96

q 0 325 7.75 7.9 21.5 80 104

r 0 330 7.86 8.1 21.5 80 100

* All fish dead, no water quality measured.

# Air stone had fallen out of tank.
C-51

Appendix C-8. Supplementary Data for Richards and Beitinger (1995)

Acclimation 

Temperature

5°C 12°C 22°C 32°C

Replicate 1 2 1 2 1 2 1 2

Sample size 30 36 30 36 36 30 33 29

pH 8.2-8.3 7.8-8.2 8.4-8.5 8.2-8.4 8.3-8.4 8.1-8.5 8.4-8.5 8.4-8.5

Hardness 

(mg/l CaCO3

)

164-180 152-166 152-168 148-170 164-174 162-172 164-168 162-172

Alkalinity 

(mg/l CaCO3)

125-140 130-140 130-140 130-140 140-145 140-145 135-140 135-145

Weights of 

minnows (g)

0.62-

3.23

0.42-2.64 0.56-2.38 0.30-1.93 0.66-

1.15

0.13-

1.55

0.26-

1.36

0.23-

1.32

Lengths of

 minnows (cm)

3.3-5.5 3.2-5.2 3.2-4.9 2.8-5.1 1.9-4.3 2.4-4.6 3.0-4.8 3.3-4.8
C-52

Appendix C-9. Data for the American River, CA, for July 1978 Through December 1980 

(data from the City of Sacramento, CA, Water Quality Laboratory; personal

communication). Units Are mg/L.

Date pH Hardness Alkalinity Ca Mg Ca:Mg Na Cl SO4

Jul-78 7.6 20 22 5.2 1.7 3.06 3.2 2.6 4

Aug-78 7.6 20 22 4.9 1.9 2.58 3.4 2.8 5

Sep-78 7.5 20 22 5.2 1.7 3.06 3.5 2.6 4

Oct-78 7.3 20 22 5 1.8 2.78 3.6 3 4

Nov-78 7.2 20 4.9 1.9 2.58 3.9 5

Dec-78

Jan-79 7.4 23 24 5.1 2.1 2.43 3.2 2.9 4

Feb-79 7.5 24 25 6.5 1.9 3.42 3 3 5

Mar-79 7.6 26 27 7.4 1.8 4.11 3.3 2.7 6

Apr-79 7.7 27 27 7.5 2 3.75 3.6 2.7 7

May-79 7.6 25 26 5.7 2.6 2.19 3.4 2.4 6

Jun-79 7.7 22 24 5.7 1.9 3.00 3.1 2.5 4

Jul-79 7.6 21 22 5.3 1.9 2.79 3 2.7 4

Aug-79 7.5 21 22 5.6 1.7 3.29 3.2 2.4 5

Sep-79 7.3 20 21 5.7 1.4 4.07 3.5 2.5 3

Oct-79 7.2 19 20 5.5 1.3 4.23 3.1 2.8 3

Nov-79

Dec-79

Jan-80 7.5 23 23 6.1 1.9 3.21 2.4 2.6 4

Feb-80 7.4 23 23 6.1 1.9 3.21 2.7 2.3 2

Mar-80 7.5 24 26 5.8 2.3 2.52 2 2.3 2

Apr-80 7.7 25 25 6.4 2.2 2.91 1.9 2.5 3

May-80 7.5 22 21 6.1 1.6 3.81 2.4 2.4 3

Jun-80 7.3 19 21 5.1 1.5 3.40 2.3 2.4 2

Jul-80 7.4 18 20 4.6 1.6 2.88 2.6 2.1 3

Aug-80 7.5 18 21 5.2 1.2 4.33 3 2.7 2

Sep-80 7.3 18 20 4.9 1.4 3.50 2.9 2.4 4

Oct-80 7.3 18 20 5 1.3 3.85 3 2.7 2

Mean 7.5 21.4 22.8 5.6 1.8 3.2 3.0 2.6 3.8

max 7.7 27.0 27.0 7.5 2.6 4.3 3.9 3.0 7.0

min 7.2 18.0 20.0 4.6 1.2 2.2 1.9 2.1 2.0
C-53

Appendix C-10. STORET Data for Minnesota Lakes and Rivers

Date pH Hardness Alkalinity Ca Mg Ca:Mg Na K Cl SO4 NO3 TOC DOC Sulfide

Embarrass River, MN

3/22/76 7 133 103 27 16 1.69 2.5 2 11 34

4/29/76 6.7 25.3 23 5.2 3 1.73 2.8 0.7 2.9 8.4 0.04 16 0.6

5/28/76 6.5 53 3.5 12

6/28/76 6.9 44 36 9.9 4.6 2.15 3.9 0.3 5 13 0.04 37

7/28/76 6.6 76 5.2 4.8 7.5

8/26/76 6.9 100 110 24 9.9 2.42 9 1 8.4 5.6 21 0.6

Means 6.8 75.58 66.83 14.26 8.38 2.00 4.55 1.00 5.93 13.42 0.04 24.67 0.60

max. 7 133 110 27 16 2.42 9 2 11 34 0.04 37 0.6

min. 6.5 25.3 23 5.2 3 1.69 2.5 0.3 2.9 5.6 0.04 16 0.6

S. Kawishiwi River, MN

10/16/75 6.4 21 14 4.9 2.1 2.33 1.3 0.4 0.5 4.4 0.01 12 0.2

11/6/75 6.9 24 19 5.5 2.5 2.20 1.2 0.4 0.6 4.1

12/11/75 39 23 10 3.4 2.94 1.4 0.4 1.5 0.2

1/9/76 6.6 29 24 6.2 3.2 1.94 1.6 0.8 2.3 7

2/4/76 6.3 24 20 5.2 2.7 1.93 1.7 0.6 0.9 6.3 0.16 16 0

3/9/76 6.9 23 23 5.7 2.2 2.59 1.5 0.5 0.9 4.9 1

4/23/76 6.6 14 8 3.4 1.3 2.62 0.9 0.4 0.7 4.8 0.2

5/25/76 6.8 16 11 4 1.5 2.67 0.9 0.4 0.7 4.8

6/25/76 6.6 16 1.1 3.3 1.8

7/23/76 6.7 19 1.2 4.4 0.5

Means 6.6 23.75 17.70 5.61 2.36 2.40 1.31 0.49 1.04 4.89 0.09 14.00 0.56

max. 6.9 39 24 10 3.4 2.94 1.7 0.8 2.3 7 0.16 16 1.8

min. 6.3 14 8 3.4 1.3 1.93 0.9 0.4 0.5 3.3 0.01 12 0

Colby Lake, MN

LCY2

6/17/96 8.5 56 33 13 5.7 2.28 4.3 1.5 6.3 22 0.25 17

6/17/96 6.8 0.25 17

6/17/96 6.9 71 33 17 7 2.43 4.3 1.4 9.4 22 18

LCY1

6/17/96 6.8 54 33 12 5.8 2.07 3.9 1.4 6.6 26 0.3 16

6/17/96 6.8 16

6/17/96 6.5 41 34 11 3.2 3.44 3.6 1.3 6.8 22 0.33 17

6/17/96 7.4 83 39 21 7.3 2.88 7.8 52 0.18

Means 7.1 55.50 33.25 13.25 5.43 2.55 4.03 1.40 7.28 23.00 0.28 16.83

max. 8.5 71 34 17 7 3.44 4.3 1.5 9.4 26 0.33 18

min. 6.5 41 33 11 3.2 2.07 3.6 1.3 6.3 22 0.25 16

Cloquet Lake, MN

7/13/76 6.4 17 11 4 1.8 2.22 1.7 7.6 0 38

Lake One, MN

10/16/75 7.2 27 21 6.9 2.3 3.00 1.2 5.6 0.02 22

Greenwood Lake, MN

7/6/76 6.7 10 15 2.8 0.7 4.00 0.1 0.3 0.2 4.2 0 11
Appendix D. Saltwater Conversion Factors for Dissolved Values
D-1

Appendix D

Saltwater Conversion Factors for Dissolved Values

February 14, 2007

U.S. Environmental Protection Agency

Office of Water

Office of Science and Technology

Washington, D.C.
D-2

ACKNOWLEDGMENTS

Larry Brooke and Tyler Linton

(primary authors)

Great Lakes Environmental Center

1295 King Avenue

Columbus, OH 43212

Jennifer Mitchell and Cindy Roberts

(authors and document coordinators)

U.S. Environmental Protection Agency

Washington, DC
D-3

Saltwater Conversion Factors for Converting Nominal or Total Copper Concentrations to

Dissolved Copper Concentrations

The U.S. EPA changed its policy in 1993 of basing water quality criteria for metals from a total

metal criteria to a dissolved metal criteria. The policy states “the use of dissolved metal to set and

measure compliance with water quality standards is the recommended approach, because dissolved metal

more closely approximates the bioavailable fraction of metal in the water column than does total

recoverable metal” (Prothro 1993). All of the criteria for metals to this date were based upon total metal

and very few data were available with dissolved concentrations of the metals. A problem was created by

the new policy of how to derive dissolved metal concentrations for studies in which this form of the

metal was not measured. The U.S. EPA attempted to develop correction factors for each metal for which

criteria exist for both fresh- and saltwater (Lussier et al. 1995; Stephan 1995). In the case of saltwater, a

correction for copper was not derived.

Several saltwater studies are available that report nominal, total, and dissolved concentrations of

copper in laboratory water (Table 1) from site-specific water effect ratio (WER) studies. These studies

show relatively consistent ratios for the nominal-to-dissolved concentrations and for the total-todissolved concentrations. Calculation of a mean ratio (conversion factor) to convert nominal and total

copper concentrations to dissolved copper permits the use of the results for critical studies without

dissolved copper measurements.

Three studies, each with multiple tests per study, were useful for deriving the conversion factors.

One study was conducted for the lower Hudson River in the New York/New Jersey Harbor (SAIC 1993).

The tests were conducted with harbor site water and with EPA Environmental Research Laboratory -

Narragansett water from Narragansett Bay, Massachusetts. Only the tests with laboratory water were

used for this exercise. Three series of 48-hour static tests were conducted with various animals. Salinity

ranged from 28 to 32 ppt during all the tests. Series 1 tests were not used to calculate ratios for dissolvedto-total or dissolved-to-nominal copper concentrations, because in many instances, concentrations of

measured copper did not increase as nominal concentrations increased. Of the series 2 tests, only the coot

clam (Mulinia lateralis) tests were successful and used to calculate ratios. Three replicate tests without

ultraviolet (UV) light present and one test with UV light present were reported with total and dissolved

copper measurements made at 0 hr and 48 hr (end) of the tests. Dissolved-to-total and dissolved-tonominal ratios were calculated for the four tests each with two time intervals. The mean ratio for the

dissolved-to-total measurements is 0.943 and the mean ratio for the dissolved-to-nominal is 0.917. A

third series of static tests was conducted by SAIC and the mussel (Mytilus sp.) test was the only

successful test. Again the tests were conducted as three replicate tests without UV light and a fourth with

UV light. The mean test ratio for dissolved-to-total copper was 0.863 and the dissolved-to-nominal mean

test ratio was 0.906.

The summer flounder (Paralichthys dentatus) was exposed to copper in laboratory water for 96

hours in a static test (CH2MHill 1999a). The water was collected from Narragansett Bay and diluted with

laboratory reverse osmosis water to dilute the solution to 22 ppt salinity. Three tests were run with

copper concentrations measured at the start of the tests as total recoverable and dissolved copper. Five

exposure concentrations were used to conduct the tests. Only the two lowest concentrations were used to

derive ratios for dissolved-to-total and dissolved-to-nominal copper mean ratios. These concentrations

were at the approximate 500 μg/L or lower concentrations, and are in the range of most copper

concentrations routinely tested in the laboratory. The mean dissolved-to-total and dissolved-to-nominal

ratios were 0.947 and 0.836, respectively.

Three 48-hour static tests were conducted with the blue mussel (Mytilus edulis) in water from the
D-4

same source and treated in the same manner as the summer flounder tests (CH2MHill 1999b). Salinity

was diluted to 20 ppt. Exposures were made at eight concentrations of copper and total and dissolved

copper concentrations were measured only at the start of the tests. Mean ratios for the dissolved-to-total

and dissolved-to-nominal copper were calculated by combining the ratios calculated for each of the test

concentrations. The mean dissolved-to-total and dissolved-to-nominal ratios were 0.979 and 0.879,

respectively.

A study was conducted by the City of San Jose, CA to develop a WER for San Francisco Bay in

which copper was used as a toxicant and the concentrations used in the laboratory exposures were

measured as total and dissolved copper (Environ. Serv. Dept., City of San Jose 1998). Mussels and the

purple sea urchin (Strongylocentrotus purpuratus) were used as the test organisms. Tests were conducted

in filtered natural sea water from San Francisco Bay that was diluted to a salinity of 28 ppt. The mussel

test was of 48-hour duration and the purple sea urchin test was of 96-hour duration. Five concentrations

of copper were used in the toxicity tests with the concentrations measured at the start of each test.

(During each test, a single concentration of copper was measured at the termination of the test and this

value was not used in the calculations.) Twenty-two tests were conducted during a 13-month period with

the mussel and two tests were conducted with the purple sea urchin. The mean dissolved-to-total and

dissolved-to-nominal ratios for the mussel tests were 0.836 and 0.785, respectively. The mean dissolvedto-total and dissolved-to-nominal ratios for the purple sea urchin were 0.883 and 0.702, respectively.

For some of the tests, control concentrations had measured concentrations of total and dissolved

copper. These values were not used to calculate ratios for dissolved-to-total and dissolved-to-nominal

copper concentrations. All mean ratios were calculated as the arithmetic mean and not as a geometric

mean of the available ratios. When the data are normally distributed, the arithmetic mean is the

appropriate measure of central tendency (Parkhurst 1998) and is a better estimator than the geometric

mean. All concentrations of copper used to calculate ratios should be time-weighted averages (Stephan

1995). In all instances of data used to calculate ratios, the concentrations were identical to time-weighted

values because either only one value was available or if two were available they were of equal weight.

Based on the information presented above the overall ratio for correcting total copper

concentrations to dissolved copper concentrations is 0.909 based upon the results of six sets of studies.

This is comparable to its equivalent factor in freshwater, which is 0.960 ± 0.037 (Stephan 1995). When it

is necessary to convert nominal copper concentrations to dissolved copper concentrations the conversion

factor is 0.838 based upon the same studies. The means of both conversion factors have standard

deviations of less than ten percent of the means (Table 1).
D-5

Table D-1. Summary of Saltwater Copper Ratios

Species

Mean Dissolved-toTotal Ratio

Mean Dissolved-toNominal Ratio Reference

Coot clam,

Mulinia lateralis 0.943 0.917 SAIC 1993

Summer flounder,

Paralichthys dentatus 0.947 0.836 CH2MHill 1999a

Blue mussel,

Mytilus sp 0.863 0.906 SAIC 1993

Blue mussel,

Mytilus edulis 0.979 0.879 CH2MHill 1999b

Blue mussel,

Mytilus sp 0.836 0.785

Environ. Serv. Dept.,

City of San Jose 1998

Purple sea urchin,

Strongylocentrotus

purpuratus

0.883 0.702 Environ. Serv. Dept.,

City of San Jose 1998

Arithmetic Mean 0.909 0.838

Standard Deviation ±0.056 ±0.082
D-6

References

CH2MH ill. 1999a. Bioassay report: Acute toxicity of copper to summer flounder (Paralichthys dentatus). Final

report prepared for U.S. Navy. November 1999. CH2MHill, Norfolk, Virginia. 26 p.

CH2MHill. 1999b. Bioassay report: Acute toxicity of copper to blue mussel (Mytilus edulis). Final report prepared

for U.S. Navy. November 1999. CH2MHill, Norfolk, Virginia. 41 p.

Environmental Services Department, City of San Jose. 1998. Development of a site-specific water quality criterion

for copper in south San Francisco Bay. Environmental Services Department, City of San Jose, San Jose/Santa Clara

Water Pollution Control Plant, 4245 Zanker Road, San Jose, CA. 171 pp. May.

Lussier, S.M., W.S. Boothman, S. Poucher, D. Champlin and A. Helmsteter. 1995. Derivation of conversion factors

for dissolved saltwater aquatic life criteria for metals. Draft report to the U.S. EPA, Office of Water. U.S. EPA,

Narragansett, RI. March 31, 1995.

Parkhurst, D.F. 1998. Arithmetic versus geometric means for environmental concentration data. Environ. Sci.

Technol./News. 32:92A-95A.

Prothro, M. 1993. Memorandum concerning “Office of Water Policy and Technical Guidance on Interpretation and

Implementation of Aquatic Life Metals Criteria.” October 1.

SAIC. 1993. Toxicity testing to support the New York/New Jersey Harbor site-specific copper criteria study. Final

Report to U.S. EPA, Office of Wastewater Enforcement and Compliance (Contract No. 68-C8-0066. Work

Assignment C-4-94). Science Applications International Corporation, Narragansett, RI.

Stephan, C.E. 1995. Derivation of conversation factors for the calculation of dissolved freshwater aquatic life criteria

for metals. Report. March 11, 1995. U.S. EPA, Duluth, MN.
Appendix E. BLM Input Data and Notes
Appendix E. BLM Table

Model Output Model Input

BLM

Data Label

Critical 

Accumulation

Hardness

(mg/L)

Temp

(oC) pH

Dissolved 

LC50 (μg/L)

DOC

(mg/L)

Humic 

Acid (%)

Ca

(mg/L)

Mg

(mg/L)

Na

(mg/L)

K

(mg/L)

SO4

(mg/L)

Cl 

(mg/L)

Alkalini

(mg/L

LUVA01S 1.1869 290 25 6.57 124.8 0.5 10 47.8602 41.47 89.821 7.178 278.4 6.5081 235 LUVA02S 2.1707 290 25 7.29 259.2 0.5 10 47.8602 41.47 89.821 7.178 278.4 6.5081 235 LUVA03S 2.0991 290 25 8.25 480 0.5 10 47.8602 41.47 89.821 7.178 278.4 6.5081 235 

CADE01F 27.6903 44.9 15 7.7 1920 1.1 10 13.1965 2.911001 1.27 0.56 3.32 1.2 42.7 CADE02F 26.6895 44.9 15 7.7 1344 1.1 10 13.1965 2.911001 1.27 0.56 3.32 1.2 42.7 

JUPL01F 0.1537 21 15 7.20 14.4 1.1 10 6.0583 1.7462 4.5302 0.7 2.8706 5.468 26 

LIVI01F 0.0570 21 15 7.2 7.68 1.1 10 6.0583 1.7462 4.5302 0.7 2.8706 5.468 26 

PHIN01F 0.4378 44.9 15 7.7 39.36 1.1 10 13.1965 2.911001 1.27 0.56 3.32 1.2 42.7 PHIN02F 0.3410 44.9 15 7.7 35.52 1.1 10 13.1965 2.911001 1.27 0.56 3.32 1.2 42.7 

ACPE01S 0.1147 96 25 8.35 25.92 0.5 10 15.8434 13.728 29.734 2.3762 92.159 2.1544 102 ACPE02S 0.1556 68 25 8.35 27.84 0.5 10 11.2224 9.724 21.061 1.6831 65.279 1.526 108 

UTIM01S 8.2925 39 23 7.4 82.56 0.5 10 6.43638 5.577 12.079 0.9653 37.439 0.8752 32.5 UTIM02S 8.0633 90 23 7.6 191.04 0.5 10 13.9716 12.11764 26.253 2.098 81.372 1.9022 65 UTIM03S 1.3555 92 25 8.1 72.96 0.5 10 29.0614 4.73839 30.798 1.6408 46.006 32.716 77 UTIM04S 1.4793 86 25 8.2 81.6 0.5 10 27.1661 4.429364 28.79 1.5338 43.005 30.583 78 UTIM05S 0.5289 90 25 8 39.36 0.5 10 28.4296 4.635381 30.129 1.6052 45.006 32.005 78 UTIM06S 1.2514 90 24 8.2 75.84 0.5 10 14.8532 12.87 13.938 1.1138 43.199 1.0099 99 UTIM07S 1.3009 90 25 7.9 69.12 0.5 10 28.4296 4.635381 30.129 1.6052 45.006 32.005 99 UTIM08S 0.7111 86 25 7.9 36.48 0.5 10 14.193 12.298 13.318 1.0643 41.279 0.965 59 

CEDU01S 0.1132 52 24.5 7.5 18.24 1.1 10 15.2833 3.371316 1.5 0.57 3.8 1.4 55 CEDU02S 0.0941 52 24.5 7.5 16.32 1.1 10 15.2833 3.371316 1.5 0.57 3.8 1.4 55 CEDU03S 0.0751 45 25 7.72 25 1.5 10 11.0991 4.2075 9.5 1.6 46 34 39.7 CEDU04S 0.0400 45 25 7.72 17 1.5 10 11.0991 4.2075 9.5 1.6 46 34 39.7 CEDU05S 0.1046 45 25 7.72 30 1.5 10 11.0991 4.2075 9.5 1.6 46 34 39.7 CEDU06S 0.0700 45 25 7.72 24 1.5 10 11.0991 4.2075 9.5 1.6 46 34 39.7 CEDU07S 0.0920 45 25 7.72 28 1.5 10 11.0991 4.2075 9.5 1.6 46 34 39.7 CEDU08S 0.1184 45 25 7.72 32 1.5 10 11.0991 4.2075 9.5 1.6 46 34 39.7 CEDU09S 0.0651 45 25 7.72 23 1.5 10 11.0991 4.2075 9.5 1.6 46 34 39.7 CEDU10S 0.0517 45 25 7.72 20 1.5 10 11.0991 4.2075 9.5 1.6 46 34 39.7 CEDU11S 0.0476 45 25 7.72 19 1.5 10 11.0991 4.2075 9.5 1.6 46 34 39.7 CEDU12S 0.0194 94.1 25 8.15 26 2 10 23.2094 8.79835 5.2449 1.6 20.054 6.1705 69.6 CEDU13S 0.0144 94.1 25 8.15 21 2 10 23.2094 8.79835 5.2449 1.6 20.054 6.1705 69.6 CEDU14S 0.0206 94.1 25 8.15 27 2 10 23.2094 8.79835 5.2449 1.6 20.054 6.1705 69.6 

E-1
Appendix E. BLM Table

Model Output Model Input

BLM

Data Label

Critical 

Accumulation

Hardness

(mg/L)

Temp

(oC) pH

Dissolved 

LC50 (μg/L)

DOC

(mg/L)

Humic 

Acid (%)

Ca

(mg/L)

Mg

(mg/L)

Na

(mg/L)

K

(mg/L)

SO4

(mg/L)

Cl 

(mg/L)

Alkalini

(mg/L

CEDU15S 0.0338 94.1 25 8.15 37 2 10 23.2094 8.79835 5.2449 1.6 20.054 6.1705 69.6 CEDU16S 0.0294 94.1 25 8.15 34 2 10 23.2094 8.79835 5.2449 1.6 20.054 6.1705 69.6 CEDU17S 0.0428 179 25 8.31 67 2.3 10 50.1069 13.12323 14.32 2.4 22.673 10.979 140.1 CEDU18S 0.0164 179 25 8.31 38 2.3 10 50.1069 13.12323 14.32 2.4 22.673 10.979 140.1 CEDU19S 0.0579 179 25 8.31 78 2.3 10 50.1069 13.12323 14.32 2.4 22.673 10.979 140.1 CEDU20S 0.0627 179 25 8.31 81 2.3 10 50.1069 13.12323 14.32 2.4 22.673 10.979 140.1 CEDU21S 0.0283 97.6 25 8 28 2 10 24.0727 9.1256 5.44 1.6 20.8 6.4 74.2 CEDU22S 0.1218 182 25 8 84 2.3 10 50.9467 13.34317 14.56 2.4 23.053 11.163 144.3 CEDU23S 0.0510 57.1 25 8.18 12.864 0.5 10 9.42352 8.1653 17.685 1.4133 54.815 1.2814 81 

CEDU24R 0.0377 80 20 7.6 5.5396825 0.5 10 13.2028 11.44 24.778 1.9801 76.799 1.7953 53 DAMA01S 0.0221 39 20 7.8 8.736 1.1 10 10.9867 2.7776 5.8136 0.7 7.9394 7.7684 51 DAMA02S 0.0315 39 20 7.8 11.232 1.1 10 10.9867 2.7776 5.8136 0.7 7.9394 7.7684 51 DAMA03S 0.0147 38 20 7.79 6.336 1.1 10 10.7129 2.7203 5.7423 0.7 7.6578 7.6406 50 DAMA04S 0.0253 38 20 7.79 9.504 1.1 10 10.7129 2.7203 5.7423 0.7 7.6578 7.6406 50 DAMA05S 0.1799 39 20 6.9 11.232 1.1 10 10.9867 2.7776 5.8136 0.7 7.9394 7.7684 30 DAMA06S 0.0786 39 20 6.9 6.432 1.1 10 10.9867 2.7776 5.8136 0.7 7.9394 7.7684 30 DAMA07S 0.0312 26 20 7.6 8.736 1.1 10 7.4273 2.0327 4.8867 0.7 4.2786 6.107 24 DAMA08S 0.0123 27 20 7.7 4.992 1.1 10 7.7011 2.09 4.958 0.7 4.5602 6.2348 24 DAMA09S 0.4278 170 20 7.8 39.552 0.5 10 27.9433 24.23527 52.507 4.1961 162.74 3.8045 115 DAMA10S 0.0443 170 20 7.8 10.08 0.5 10 27.9433 24.23527 52.507 4.1961 162.74 3.8045 115 DAMA11S 0.1330 170 20 7.8 19.776 0.5 10 27.9433 24.23527 52.507 4.1961 162.74 3.8045 115 DAMA12S 0.0990 170 20 7.8 16.608 0.5 10 27.9433 24.23527 52.507 4.1961 162.74 3.8045 115 DAMA13S 0.9670 170 20 7.8 67.872 0.5 10 27.9433 24.23527 52.507 4.1961 162.74 3.8045 115 DAMA14S 0.2716 170 20 7.8 30.048 0.5 10 27.9433 24.23527 52.507 4.1961 162.74 3.8045 115 DAMA15S 0.0160 109.9 21 6.93 6.816 2.4 10 40.0 2.43 85.1 1.23 10 106 12.5 DAMA16S 0.0298 109.9 21 6.93 15.744 3.4 10 40.0 2.43 85.1 1.23 10 106 12.5 DAMA17S 0.0393 109.9 21 7.43 38.304 3.4 10 40.0 2.43 85.1 1.23 10 106 13.875 DAMA18S 0.0219 109.9 21 7.43 17.952 2.4 10 40.0 2.43 85.1 1.23 10 106 13.875 DAMA19S 0.0111 109.9 21 7.82 18.144 2.4 10 40.0 2.43 85.1 1.23 10 106 14.5 DAMA20S 0.0189 109.9 21 7.82 38.112 3.4 10 40.0 2.43 85.1 1.23 10 106 14.5 DAMA21S 0.0898 109.9 21 6.93 44.16 4.4 10 40.0 2.43 85.1 1.23 10 106 12.5 DAMA22S 0.1076 109.9 21 6.93 69.024 6.1 10 40.0 2.43 85.1 1.23 10 106 12.5 DAMA23S 0.0458 109.9 21 7.43 54.912 4.4 10 40.0 2.43 85.1 1.23 10 106 13.875 DAMA24S 0.0288 109.9 21 7.82 65.088 4.4 10 40.0 2.43 85.1 1.23 10 106 14.5 

E-2
Appendix E. BLM Table

Model Output Model Input

BLM

Data Label

Critical 

Accumulation

Hardness

(mg/L)

Temp

(oC) pH

Dissolved 

LC50 (μg/L)

DOC

(mg/L)

Humic 

Acid (%)

Ca

(mg/L)

Mg

(mg/L)

Na

(mg/L)

K

(mg/L)

SO4

(mg/L)

Cl 

(mg/L)

Alkalini

(mg/L

DAMA25S 0.1143 52 18.2 7.8 24.96 1.1 10 14 3.5 12 2.9 23 11 45 DAMA26S 0.0917 105 20.3 7.9 28.8 1.1 10 29 6.8 29 5.3 57 21 79 DAMA27S 0.1053 106 19.7 8.1 36.48 1.1 10 29 6.8 29 5.3 57 21 82 DAMA28S 0.1538 207 19.9 8.3 66.24 1.1 10 58 13 62 8.2 127 40 166 DAMA29S 0.0062 7.1 24 8.55 4.608 0.5 10 1.15182 1.027387 3.5102 2.8052 6.8159 2.5434 56 DAMA30S 0.2536 20.6 24 6.97 7.104 0.5 10 3.39973 2.9458 2.5478 2.1356 19.776 1.9363 60 DAMA31S 0.0119 23 24 8.52 6.24 0.5 10 3.79581 3.289 2.8446 2.3845 22.08 2.1619 64 

DAPC01S 0.0087 48 18 8.03 10.944 2.288 10 14.1077 3.111984 1.36 0.57 3.55 1.25 42 DAPC02S 0.0052 48 18 8.03 8.6976 2.816 10 14.1077 3.111984 1.36 0.57 3.55 1.25 42 DAPC03S 0.0043 48 18 8.01 6.9504 2.728 10 14.1077 3.111984 1.36 0.57 3.55 1.25 44 DAPC04S 0.0057 44 18 8.04 10.368 3.08 10 12.932 2.852652 1.24 0.57 3.25 1.15 42 DAPC05S 0.0879 31 18 6.66 53.184 12.2094 10 7.37407 3.063455 1.6792 0.5 6.3292 1.2917 27 DAPC06S 0.0490 29 18 6.97 53.088 11.3373 10 6.89832 2.865813 1.5708 0.5 5.9208 1.2083 27 DAPC07S 0.0285 28 18 7.2 51.168 11.3373 10 6.66045 2.766992 1.5167 0.5 5.7167 1.1667 22 DAPC08S 0.0268 88 18 7.01 93.312 24.4188 10 20.9464 8.5194 16.466 1.8787 22.629 18.986 20 DAPC09S 0.0187 100 18 7.55 191.04 29.6514 10 23.9296 9.4686 21.207 2.1631 25.98 23.28 20 DAPC10S 0.0701 82 18 6.99 204.48 27.9072 10 19.4548 8.0448 14.095 1.7365 20.953 16.84 18 DAPC11S 0.0460 84 18 7.01 158.4 27.9072 10 19.952 8.203 14.885 1.7839 21.512 17.555 17 DAPC12S 0.0100 16 18 7.39 34.08 11.6124 10 4.13844 1.379481 0.16 0.3 6.72 0.32 11 DAPC13S 0.0137 151 18 7.76 75.648 12.5801 10 36.7872 14.39533 10.786 1.4 62.018 19.684 44 DAPC14S 0.0053 96 18 8.1 108.48 27.0956 10 22.0888 9.939946 6.8571 1.4 19.911 4.2667 91 DAPC15S 0.0137 26 18 7.24 73.344 24.1925 10 7.37925 1.844812 0.26 0.3 11.624 2.6 4 DAPC16S 0.0564 84 18 7.08 81.312 12.5801 10 20.4644 8.008 6 1.4 34.5 10.95 13 DAPC17S 0.0633 92 18 7.22 176.64 20.3217 10 22.4134 8.770667 6.5714 1.4 37.786 11.993 19 DAPC18S 0.0056 47 18 8.03 8.928 2.728 10 13.8137 3.047151 1.33 0.57 3.47 1.23 42.5 DAPC19S 0.0119 97 18 8.03 17.088 2.728 10 34 2.9 1.3 0.57 51.3 1.2 42.5 DAPC20S 0.0160 147 18 8.03 22.752 2.728 10 54 2.9 1.3 0.57 99.3 1.2 42.5 DAPC21S 0.0168 247 18 8.03 26.208 2.728 10 94 2.9 1.3 0.57 147.3 1.2 42.5 DAPC22S 0.0171 97 18 8.03 24.192 2.728 10 13.6 15.2 1.3 0.57 51.3 1.2 42.5 DAPC23S 0.0155 147 18 8.03 24.096 2.728 10 13.6 27.5 1.3 0.57 99.3 1.2 42.5 DAPC24S 0.0133 247 18 8.03 24.096 2.728 10 13.6 51.9 1.3 0.57 147.3 1.2 42.5 

SCSP01S 0.1034 52 24.5 7.5 17.28 1.1 10 15.2833 3.371316 1.47 0.57 3.84 1.36 55 GAPS01F 0.1153 44.9 15 7.7 21.12 1.1 10 13.1965 2.911001 1.27 0.57 3.32 1.17 42.7 GAPS02F 0.0888 44.9 15 7.7 18.24 1.1 10 13.1965 2.911001 1.27 0.57 3.32 1.17 42.7 

E-3
Appendix E. BLM Table

Model Output Model Input

BLM

Data Label

Critical 

Accumulation

Hardness

(mg/L)

Temp

(oC) pH

Dissolved 

LC50 (μg/L)

DOC

(mg/L)

Humic 

Acid (%)

Ca

(mg/L)

Mg

(mg/L)

Na

(mg/L)

K

(mg/L)

SO4

(mg/L)

Cl 

(mg/L)

Alkalini

(mg/L

HYAZ01S 0.1511 290 25 6.23 16.32 0.5 10 47.8602 41.47 89.821 7.178 278.4 6.5081 235 HYAZ02S 0.1074 290 25 7.51 23.04 0.5 10 47.8602 41.47 89.821 7.178 278.4 6.5081 235 HYAZ03S 0.2392 290 25 8.38 83.52 0.5 10 47.8602 41.47 89.821 7.178 278.4 6.5081 235 HYAZ04S 0.0794 20.5 21 7.15 23.328 2.8 10 5.1 1.9 5.3 0.8 9.3 10.0 6.7 HYAZ05S 0.0768 20.5 21 7.15 22.848 2.8 10 5.1 1.9 5.3 0.8 9.3 10.0 6.7 HYAZ06S 0.2314 20.6 21 7.14 7.872 0.5 10 5.3 1.8 5.5 0.8 7.0 9.7 11.0 HYAZ07S 0.3312 20.6 21 7.14 9.6 0.5 10 5.3 1.8 5.5 0.8 7.0 9.7 11.0 

ACLY01S 29.5658 42 18.5 7.0 7968 1.1 10 12.3442 2.722986 1.3 0.57 3.4 1.2 47 

CHDE01S 25.2731 44 20 7.40 709.44 0.5 10 6.99 6.06 13.1 1.05 40.7 0.951 32.5 

SCPL01S 2.9865 167 22 7.6 153.6 0.5 10 27.5609 23.881 51.724 4.1335 160.32 3.7478 115 

ONAP01S 0.9139 169 12 8 67.2 0.5 10 27.891 24.167 52.344 4.183 162.24 3.7927 117 

ONCL01S 1.0007 169 12 8.1 76.8 0.5 10 27.891 24.167 52.344 4.183 162.24 3.7927 117 ONCL02S 0.5538 169 12 8.25 57.6 0.5 10 27.891 24.167 52.344 4.183 162.24 3.7927 117 

ONCL03F 2.8512 205 13.7 7.73 367 3.3 10 49.8 19.6 4 0.64 10 0.44 178 ONCL04F 1.5731 69.9 13.7 8.54 186 1.5 10 18.4 5.8 1.405 0.2248 3.5126 0.1546 174 ONCL05F 0.4400 18 13.7 8.07 36.8 0.75 10 4.8 1.5 0.3618 0.0579 0.9045 0.0398 183 ONCL06F 1.9714 204 13.7 7.61 232 3.3 10 64.7 10.3 4.1005 0.6561 10.251 0.4511 77.9 ONCL07F 5.2514 83 13.7 7.4 162 1.7 10 20.4 7.8 1.6683 0.2669 4.1709 0.1835 70 ONCL08F 1.2778 31.4 13.7 8.32 73.6 0.94 10 7.9 2.7 0.6312 0.101 1.5779 0.0694 78.3 ONCL09F 0.3591 160 13.7 7.53 91 2.8 10 57.5 4.0 3.2161 0.5146 8.0402 0.3538 26.0 ONCL10F 0.3318 74.3 13.7 7.57 44.4 1.5 10 24.7 3.1 1.4935 0.239 3.7337 0.1643 22.7 ONCL11F 0.1192 26.4 13.7 7.64 15.7 0.87 10 6.0 2.8 0.5307 0.0849 1.3266 0.0584 20.1 

ONGO01F 1.3932 83.1 7.15 7.63 137.28 2.58 10 22.3428 6.313221 10.259 7.5024 25.1 9.994 62.5 ONGO02F 0.3615 83.1 7.15 7.63 83.52 2.58 10 22.3428 6.313221 10.259 7.5024 25.1 9.994 62.5 ONGO03F 3.5018 83.1 7.15 7.63 191.04 2.58 10 22.3428 6.313221 10.259 7.5024 25.1 9.994 62.5 

ONKI01R 4.9807 33 13.5 7.29 157.44 2.496 10 8.77741 2.698479 7.3188 1.15 6.1426 6.8124 29 

ONKI02F 0.4054 25 12 7.30 31.68 1.3 10 6.8 1.8 5.0 0.6 4.2 6 24 ONKI03F 0.9203 20 9.4 7.29 44.16 1.3 10 5.7845 1.6889 4.4589 0.7 2.589 5.3402 22 ONKI04F 0.1617 31.1 13.3 7.30 49 3.2 10 8.01999 2.695987 5.12 0.653 4 4.5 29.6 ONKI05F 0.1736 31.1 13.3 7.30 51 3.2 10 8.01999 2.695987 5.12 0.653 4 4.5 29.6 ONKI06F 0.1461 31.6 15.7 7.50 58 3.2 10 8.14893 2.739331 5.12 0.653 3.5 4.2 30.4 ONKI07F 0.4829 31 15.3 7.20 78 3.2 10 7.99421 2.687318 5.12 0.653 2.3 3.1 29.7 

ONMY01S 1.3925 169 12 8.2 105.6 0.5 10 27.891 24.167 52.344 4.183 162.24 3.7927 117 ONMY02S 0.5765 169 12 7.95 48 0.5 10 27.891 24.167 52.344 4.183 162.24 3.7927 117 

E-4
Appendix E. BLM Table

Model Output Model Input

BLM

Data Label

Critical 

Accumulation

Hardness

(mg/L)

Temp

(oC) pH

Dissolved 

LC50 (μg/L)

DOC

(mg/L)

Humic 

Acid (%)

Ca

(mg/L)

Mg

(mg/L)

Na

(mg/L)

K

(mg/L)

SO4

(mg/L)

Cl 

(mg/L)

Alkalini

(mg/L

ONMY03S 0.7648 169 12 7.95 57.6 0.5 10 27.891 24.167 52.344 4.183 162.24 3.7927 117 

ONMY04R 0.1249 44.1 11.5 7.7 40 2 10 9.07 4.1 4.75 1.02 3.3 1.56 49.7 ONMY05R 0.0917 44.6 11.5 7.8 19 0.99 10 7.37 6.1 6.24 0.8 1.31 3.82 53.1 ONMY06R 0.0376 38.7 12 7.62 3.4 0.33 10 2.37 8.65 13.7 0.15 0.36 20.3 40 ONMY07R 0.1465 39.3 12 7.61 8.1 0.36 10 14.1 1.8 13.2 0.1 0.36 19.9 41.7 ONMY08R 0.1881 89.5 12 8.21 17.2 0.345 10 15 11.85 10.05 1 0.36 6.73 97.5 ONMY09R 0.5172 89.67 12 8.15 32 0.345 10 28.9 3.15 32.5 0.5 0.36 45.2 97.25 

ONMY10F 0.3824 23 12.2 7.1 26.88 1.4 10 6.1 1.8 4.4 0.4 5.8 6 22 ONMY11F 0.1589 23 12.2 7.1 16.32 1.4 10 6.1 1.8 4.4 0.4 5.8 6 22 ONMY12F 0.1059 23 12.2 7.4 17.28 1.3 10 6.8 1.8 5.0 0.6 4.2 6 22 ONMY13F 0.4633 23 12.2 7.1 27.84 1.3 10 6.8 1.8 5.0 0.6 4.2 6 22 ONMY14F 0.4998 194 12.8 7.84 169 3.3 10 55.1 13.7 4 0.64 10 0.44 174 ONMY15F 0.1118 194 12.8 7.84 85.3 3.3 10 55.1 13.7 4 0.64 10 0.44 174 ONMY16F 0.1069 194 12.8 7.84 83.3 3.3 10 55.1 13.7 4 0.64 10 0.44 174 ONMY17F 0.1627 194 12.8 7.84 103 3.3 10 55.1 13.7 4 0.64 10 0.44 174 ONMY18F 1.5525 194 12.8 7.84 274 3.3 10 55.1 13.7 4 0.64 10 0.44 174 ONMY19F 0.2605 194 12.8 7.84 128 3.3 10 55.1 13.7 4 0.64 10 0.44 174 ONMY20F 0.9538 194 12.8 7.84 221 3.3 10 55.1 13.7 4 0.64 10 0.44 174 ONMY21F 0.4717 194 12.8 7.84 165 3.3 10 55.1 13.7 4 0.64 10 0.44 174 ONMY22F 0.7244 194 12.8 7.84 197 3.3 10 55.1 13.7 4 0.64 10 0.44 174 ONMY23F 4.6605 194 12.8 7.84 514 3.3 10 55.1 13.7 4 0.64 10 0.44 174 ONMY24F 1.1894 194 12.8 7.84 243 3.3 10 55.1 13.7 4 0.64 10 0.44 174 ONMY25F 0.0613 9.2 15.5 6.96 2.688 0.5 10 2.3 0.7 2 0.2 4.6 2.1 11 ONMY26F 0.3626 31 15.3 7.2 68 3.2 10 7.99421 2.687318 5.12 0.653 2.3 3.1 29.7 ONMY27F 0.0770 36.1 11.4 7.6 18 1.31 10 4.03 7.13 1.56 0.26 1.49 0.88 36.6 ONMY28F 0.8944 36.2 11.5 6.1 12 1.36 10 3.93 7.27 1.57 0.28 1.47 0.87 8.5 ONMY29F 0.5568 20.4 11.7 7.5 5.7 0.15 10 3.13 2.77 2.62 0.25 0.36 1.48 23 ONMY30F 0.2504 45.2 11.7 7.7 35 1.23 10 9.7 4.43 5.33 0.97 3.41 1.47 50 ONMY31F 1.1775 45.4 11.8 6.3 18 1.22 10 9.7 4.43 5.02 0.98 3.37 1.37 10.9 ONMY32F 0.5318 41.9 12.3 7.9 17 0.33 10 6.6 5.97 5.89 0.63 1.11 3.37 48.3 ONMY33F 1.2884 214 7.64 7.94 96.96 0.27 10 49.4 24.1 10.3 1.75 18.9 5.28 198 ONMY34F 3.8957 220 7.74 7.92 295.68 0.36 10 51.2 25.5 8.36 2.1 24 4.64 197 ONMY35F 4.4437 105 7.77 7.82 89.28 0.1 10 23.1 11.8 3.54 3.22 17.1 2.91 94.1 ONMY36F 1.9096 98.2 8.49 7.89 34.464 0.045 10 22.3 11.2 3.58 0.9 11.5 2.85 87.9 

E-5
Appendix E. BLM Table

Model Output Model Input

BLM

Data Label

Critical 

Accumulation

Hardness

(mg/L)

Temp

(oC) pH

Dissolved 

LC50 (μg/L)

DOC

(mg/L)

Humic 

Acid (%)

Ca

(mg/L)

Mg

(mg/L)

Na

(mg/L)

K

(mg/L)

SO4

(mg/L)

Cl 

(mg/L)

Alkalini

(mg/L

ONMY37F 1.7297 104 16.3 7.83 52.224 0.28 10 22.4 11.4 3.76 2.72 12.4 3.01 97.6 

ONNE01F 3.1060 83.1 7.15 7.63 182.4 2.58 10 22.3428 6.313221 10.259 7.5024 25.1 9.994 62.5 ONNE02F 3.5466 83.1 7.15 7.63 192 2.58 10 22.3428 6.313221 10.259 7.5024 25.1 9.994 62.5 ONNE03F 0.5132 83.1 7.15 7.63 96 2.58 10 22.3428 6.313221 10.259 7.5024 25.1 9.994 62.5 ONNE04F 0.6617 83.1 7.15 7.63 105.6 2.58 10 22.3428 6.313221 10.259 7.5024 25.1 9.994 62.5 ONNE05F 1.0574 83.1 7.15 7.63 124.8 2.58 10 22.3428 6.313221 10.259 7.5024 25.1 9.994 62.5 ONNE06F 1.6007 83.1 7.15 7.63 144 2.58 10 22.3428 6.313221 10.259 7.5024 25.1 9.994 62.5 ONNE07F 4.0021 83.1 7.15 7.63 201.6 2.58 10 22.3428 6.313221 10.259 7.5024 25.1 9.994 62.5 ONNE08F 2.2920 83.1 7.15 7.63 163.2 2.58 10 22.3428 6.313221 10.259 7.5024 25.1 9.994 62.5 ONNE09F 3.1060 83.1 7.15 7.63 182.4 2.58 10 22.3428 6.313221 10.259 7.5024 25.1 9.994 62.5 ONNE10F 5.4103 83.1 7.15 7.63 230.4 2.58 10 22.3428 6.313221 10.259 7.5024 25.1 9.994 62.5 

ONTS01F 0.2050 23 12.2 7.4 24.96 1.3 10 6.8 1.8 5.0 0.6 4.2 6 22 ONTS02F 0.1161 23 12.2 7.4 18.24 1.3 10 6.8 1.8 5.0 0.6 4.2 6 22 ONTS03F 0.7109 23 12.2 7.1 36.48 1.4 10 6.1 1.8 4.4 0.4 5.8 6 22 ONTS04F 0.3750 23 12.2 7.1 24.96 1.3 10 6.8 1.8 5.0 0.6 4.2 6 22 ONTS05F 0.3517 13 12 7.15 9.792 0.5 10 2.14546 1.859 4.0264 0.3218 12.48 0.2917 12 ONTS06F 0.8340 46 12 7.55 23.136 0.5 10 7.59162 6.578 14.247 1.1386 44.159 1.0323 35 ONTS07F 0.9241 182 12 8.12 79.2 0.5 10 30.0364 26.026 56.37 4.5048 174.72 4.0844 125 ONTS08F 0.3954 359 12 8.49 123.264 0.5 10 59.2477 51.337 111.19 8.8858 344.64 8.0566 243 ONTS09F 1.1161 36.6 12 7.71 7.4 0.055 10 6.36 4.73 4.84 0.22 0.94 2.79 40.8 ONTS10F 0.8313 34.6 12 7.79 12.5 0.19 10 7.82 3.17 9.98 0.11 0.73 8.34 40.6 ONTS11F 0.8622 38.3 12 7.71 14.3 0.24 10 6.33 5.1 5.27 0.6 0.99 2.96 43.6 ONTS12F 1.7785 35.7 12 7.74 18.3 0.17 10 8.15 3.38 10 0.37 0.76 9.1 43.3 

SACO01F 2.9901 214 7.64 7.94 218.88 0.27 10 49.4 24.1 10.3 1.75 18.9 5.28 198 SACO02F 2.6420 220 7.74 7.92 198.72 0.36 10 51.2 25.5 8.36 2.1 24 4.64 197 SACO03F 3.2456 105 7.77 7.82 63.936 0.1 10 23.1 11.8 3.54 3.22 17.1 2.91 94.1 SACO04F 2.6405 98.2 8.49 7.89 48 0.045 10 22.3 11.2 3.58 0.9 11.5 2.85 87.9 SACO05F 3.0680 104 16.3 7.83 85.44 0.28 10 22.4 11.4 3.76 2.72 12.4 3.01 97.6 

ACAL01F 9.7513 54 10.5 7.3 137.28 1.1 10 15.0937 3.6371 6.8831 0.7 12.163 9.6854 43 

GIEL01S 2.6186 173 22 8.05 192 0.5 10 28.5511 24.739 53.583 4.282 166.08 3.8824 117 

NOCR01F 29.9790 72.2 25 7.50 81216 1.5 10 17.8079 6.7507 15.26 1.6 73.841 54.15 42.5 

PIPR01S 11.3981 103 22 7.4 297.6 0.5 10 28.4667 7.773195 27.778 2.6358 29.602 53.021 65 PIPR02S 4.9570 103 22 7.4 115.2 0.5 10 28.4667 7.773195 27.778 2.6358 29.602 53.021 65 PIPR03S 9.4256 263 22 7.4 374.4 0.5 10 72.6868 19.84806 36.487 3.4623 77.901 130.77 65 

E-6
Appendix E. BLM Table

Model Output Model Input

BLM

Data Label

Critical 

Accumulation

Hardness

(mg/L)

Temp

(oC) pH

Dissolved 

LC50 (μg/L)

DOC

(mg/L)

Humic 

Acid (%)

Ca

(mg/L)

Mg

(mg/L)

Na

(mg/L)

K

(mg/L)

SO4

(mg/L)

Cl 

(mg/L)

Alkalini

(mg/L

PIPR04S 1.2005 52 24.5 7.4 52.8 1.1 10 15.2833 3.371316 1.47 0.57 3.84 1.36 55 PIPR05S 3.0479 52 24.5 7.4 81.6 1.1 10 15.2833 3.371316 1.47 0.57 3.84 1.36 55 PIPR06S 0.1314 290 25 6.27 14.4 0.5 10 47.8602 41.47 89.821 7.178 278.4 6.5081 235 PIPR07S 0.3064 290 25 7.14 42.24 0.5 10 47.8602 41.47 89.821 7.178 278.4 6.5081 235 PIPR08S 0.5392 290 25 8.6 192 0.5 10 47.8602 41.47 89.821 7.178 278.4 6.5081 235 PIPR09S 0.0890 19 22 7.06 4.6272 0.6 10 4.9 1.64 3.7 0.78 9.6 5.8 11.17 PIPR10S 0.2665 19.5 22 7.25 7.872 0.4 10 5.2 1.64 5.36 0.79 2.45 8.6 12.7 PIPR11S 0.5716 16.5 22 6.36 30.3072 3.3 10 4.1 1.54 2.82 0.76 9.4 4.7 8.46 PIPR12S 0.2950 17 22 6.42 20.2176 3.1 10 4.2 1.56 2.74 0.74 7.4 4.6 3.4 PIPR13S 0.4162 19 22 6.38 34.5312 4.3 10 5 1.62 7.04 0.72 10.2 12.2 7.83 PIPR14S 0.2640 17 22 7.15 57.4368 3.4 10 4.2 1.54 2.9 1 7.4 4.7 8.74 PIPR15S 0.0477 17 22 7.16 4.6368 0.8 10 4.5 1.46 2.68 0.78 10.9 3.8 9.3 PIPR16S 0.1770 17.5 22 7.13 67.4688 5.1 10 4.6 1.48 2.62 0.77 10.5 3.5 8.95 PIPR17S 0.0787 18.5 22 7.06 80.2464 10.5 10 5 1.54 2.64 0.8 10.7 3.5 8.29 PIPR18S 0.1907 18.5 22 6.90 174.72 15.6 10 4.9 1.5 3.54 0.99 7 5.2 9.52 PIPR19S 3.2305 173 22 8.25 278.4 0.5 10 28.5511 24.739 53.583 4.282 166.08 3.8824 117 PIPR20S 7.4512 173 22 8.1 604.8 0.5 10 28.5511 24.739 53.583 4.282 166.08 3.8824 117 PIPR21S 4.8297 173 22 8.15 384 0.5 10 28.5511 24.739 53.583 4.282 166.08 3.8824 117 PIPR22S 7.6122 173 22 7.3 374.4 0.5 10 28.5511 24.739 53.583 4.282 166.08 3.8824 117 PIPR23S 7.2327 166 5 8.05 432 0.5 10 27.3959 23.738 51.415 4.1088 159.36 3.7253 132.5 PIPR24S 3.4469 159 12 8.35 285.12 0.5 10 26.2406 22.737 49.247 3.9355 152.64 3.5682 135 PIPR25S 2.8678 168 22 8.3 298.56 0.5 10 27.7259 24.024 52.034 4.1583 161.28 3.7702 142.5 PIPR26S 3.3686 167 32 8.45 492.48 0.5 10 27.5609 23.881 51.724 4.1335 160.32 3.7478 140 PIPR27S 0.5950 45.54059 22 7.93 53.958366 1.1 10 13.4911 2.888065 1.6093 0.391 3.362 1.4181 42.0374PIPR28S 4.0104 45.54059 22 7.93 165.17867 1.1 10 13.4911 2.888065 91.27 0.391 3.362 143.23 42.0374PIPR29S 0.7241 44.53969 22 7.98 59.464322 1.1 10 13.1946 2.824591 1.6093 0.391 3.362 1.4181 42.0374PIPR30S 4.0805 44.53969 22 7.98 146.45842 1.1 10 13.1946 2.824591 45.98 0.391 3.362 72.324 44.0392PIPR31S 1.8188 44.53969 22 7.99 82.038741 1.1 10 13.1946 2.824591 1.6093 0.391 3.362 1.4181 42.5379PIPR32S 4.9213 45.54059 22 7.96 124.4346 1.1 10 13.4911 2.888065 1.6093 0.391 3.362 36.871 43.0383PIPR33S 3.9367 45.04014 22 7.79 103.759 1.1 10 13.3428 2.856328 1.6093 0.391 3.362 1.4181 46.0410PIPR34S 5.7875 45.04014 22 7.81 167.3225 1.1 10 13.3428 2.856328 47.589 0.391 99.42 1.4181 46.0410PIPR35S 3.2914 138.1231 22 7.785 120.015 1.1 10 12.892 25.75825 1.6093 0.391 3.362 72.324 43.0383PIPR36S 5.7959 151.1347 22 7.78 169.418 1.1 10 14.1065 28.18476 1.6093 0.391 99.42 1.4181 43.0383PIPR37S 3.4870 138.1231 22 8.02 268.224 1.1 10 12.892 25.75825 1.6093 0.391 3.362 1.4181 149.132

E-7
Appendix E. BLM Table

Model Output Model Input

BLM

Data Label

Critical 

Accumulation

Hardness

(mg/L)

Temp

(oC) pH

Dissolved 

LC50 (μg/L)

DOC

(mg/L)

Humic 

Acid (%)

Ca

(mg/L)

Mg

(mg/L)

Na

(mg/L)

K

(mg/L)

SO4

(mg/L)

Cl 

(mg/L)

Alkalini

(mg/L

PIPR38S 9.2068 139.124 22 7.775 242.443 1.1 10 51.1778 2.779812 1.6093 0.391 99.42 1.4181 43.0383PIPR39S 4.7038 47.04192 22 7.78 113.3475 1.1 10 13.4268 4.010325 1.6093 0.391 3.362 1.4181 43.0383PIPR40S 3.1754 37.033 22 7.785 77.8764 0.88 10 11.022 3.281175 2.9887 0.391 3.362 1.4181 43.0383PIPR41S 5.3335 60.05352 22 7.795 128.016 1.1 10 15.2304 5.954725 1.6093 0.391 17.771 1.4181 43.0383PIPR42S 6.4718 76.06779 22 7.8 151.13 1.1 10 18.8376 7.413025 1.6093 0.391 32.179 1.7727 42.0374PIPR43S 6.4642 103.0919 22 7.805 166.624 1.1 10 25.05 10.2081 2.0691 0.391 60.036 1.7727 43.0383PIPR44S 7.0015 103.0919 22 7.78 163.83 1.1 10 32.064 4.010325 1.8392 0.391 58.115 1.7727 40.0356PIPR45S 5.9820 107.0954 22 7.79 157.48 1.1 10 18.2364 15.43368 1.6093 0.391 61.957 1.7727 43.0383PIPR46S 7.4331 134.1195 22 7.8 199.7075 1.1 10 32.2644 13.00318 1.6093 0.391 88.854 1.7727 43.0383PIPR47S 6.0725 45.04014 22 7.815 128.524 1.1 10 14.028 2.18745 1.3794 0.391 3.362 1.0636 41.0365PIPR48S 7.2713 46.04103 22 7.82 150.876 1.1 10 14.028 2.18745 6.2072 1.5639 5.7635 7.0906 42.0374PIPR49S 5.4175 45.04014 22 7.82 131.064 1.1 10 14.028 2.18745 15.173 1.5639 10.566 15.245 41.0365PIPR50S 6.2395 45.04014 22 7.81 160.2105 1.1 10 14.2284 2.18745 35.174 1.5639 21.613 36.162 41.0365PIPR51S 6.2194 44.03925 22 7.82 182.88 1.1 10 15.03 2.18745 62.992 1.5639 40.825 70.906 40.0356PIPR52S 4.9667 45.04014 22 7.81 180.848 1.1 10 14.4288 2.18745 101.39 1.9549 59.076 107.78 41.0365PIPR53S 6.1183 46.04103 22 7.81 176.784 1.1 10 14.2284 2.18745 57.015 19.158 40.825 71.97 42.0374PIPR54S 5.7931 189.1686 22 7.82 188.9125 1.1 10 55.11 15.79825 1.6093 0.782 152.25 1.0636 42.0374PIPR55S 5.2814 46.04103 22 7.865 125.603 1.1 10 14.6292 3.15965 1.3794 0.391 3.362 1.0636 42.0374PIPR56S 3.8765 75.0669 22 7.87 117.348 1.1 10 24.4488 5.954725 1.3794 0.391 30.739 1.0636 41.0365PIPR57S 3.7460 46.04103 22 7.865 114.554 1.1 10 14.4288 3.15965 19.771 0.391 12.488 18.436 41.0365PIPR58S 3.8963 74.06601 22 7.85 126.492 1.1 10 24.4488 6.07625 18.392 0.391 38.903 18.436 42.0374PIPR59S 5.1820 133.1186 22 7.85 172.72 1.1 10 41.082 11.6664 18.392 0.391 98.94 18.436 42.0374PIPR60S 5.0050 76.06779 22 7.85 167.3225 1.1 10 24.048 6.07625 47.589 0.782 58.115 52.116 43.0383PIPR61S 6.3379 134.1195 22 7.84 226.695 1.1 10 40.8816 11.6664 49.198 0.782 118.63 51.052 43.0383PIPR62S 6.5522 52.04638 22 7.96 84.201 0.3 10 12.024 4.13185 1.6093 0.391 10.566 1.7727 42.0374PIPR63S 7.7846 51.04549 22 7.96 97.79 0.3 10 11.2224 3.8888 2.7588 0.782 10.566 3.5453 41.0365PIPR64S 5.4254 50.0446 22 7.945 70.0786 0.3 10 11.022 3.767275 5.9773 1.5639 12.007 8.1542 41.0365PIPR65S 5.7632 51.04549 22 7.965 81.5848 0.3 10 11.2224 3.8888 11.955 2.3459 15.369 15.245 42.0374PIPR66S 5.0152 51.04549 22 7.96 77.4319 0.3 10 11.2224 3.767275 23.22 3.1279 21.613 30.135 41.0365PIPR67S 5.9195 53.04728 22 7.97 110.871 0.3 10 11.2224 3.767275 46.899 4.6918 33.62 59.207 41.5370PIPR68S 5.4017 53.04728 22 7.96 151.892 0.3 10 11.6232 3.8888 117.94 7.0377 68.201 141.81 42.0374PIPR69S 4.1225 52.04638 22 7.94 175.26 0.3 10 11.4228 3.767275 236.79 10.948 128.24 279.72 43.0383PIPR70S 6.6575 47.04192 25 7.82 145.288 1.1 10 13.9359 2.983276 1.6093 0.391 3.362 1.4181 42.5379PIPR71S 4.6725 47.04192 20 7.82 111.76 1.1 10 13.9359 2.983276 1.6093 0.391 3.362 1.4181 43.0383

E-8
Appendix E. BLM Table

Model Output Model Input

BLM

Data Label

Critical 

Accumulation

Hardness

(mg/L)

Temp

(oC) pH

Dissolved 

LC50 (μg/L)

DOC

(mg/L)

Humic 

Acid (%)

Ca

(mg/L)

Mg

(mg/L)

Na

(mg/L)

K

(mg/L)

SO4

(mg/L)

Cl 

(mg/L)

Alkalini

(mg/L

PIPR72S 2.3613 47.04192 15 7.82 79.1845 1.1 10 13.9359 2.983276 1.6093 0.391 3.362 1.4181 42.5379PIPR73S 1.1782 47.04192 10 7.82 60.0075 1.1 10 13.9359 2.983276 1.6093 0.391 3.362 1.4181 42.5379PIPR74S 7.6860 140.1249 22 8.03 370.078 0.3 10 29.058 12.03098 25.059 4.3008 60.036 25.881 98.0874PIPR75S 10.9585 88.0785 22 7.965 292.1 0.3 10 19.038 7.04845 14.943 2.7369 37.943 17.017 63.0561PIPR76S 7.9470 59.05263 22 7.89 101.473 0.3 10 12.024 4.61795 9.1959 0.782 23.054 9.9268 39.0347PIPR77S 6.9448 41.03657 22 7.825 62.5094 0.3 10 8.2164 3.038125 7.5866 2.7369 13.928 6.3815 29.0258PIPR78S 5.9976 27.02408 22 7.745 42.0624 0.3 10 5.6112 1.822875 4.598 2.3459 8.6452 4.2544 23.0205PIPR79S 9.0570 43.03836 22 7.885 172.466 1.1 10 10.4208 2.67355 1.6093 0.782 2.8817 1.4181 42.0374PIPR80S 0.7034 25.0223 22 7.565 12.4333 0.3 10 6.68596 2.02764 3.4485 1.1729 4.3226 4.9634 16.0142PIPR81S 7.0672 107.0954 22 8.105 271.272 0.3 10 28.6924 8.631893 14.254 1.9549 19.212 16.308 80.0713PIPR82S 4.9660 87.0776 22 7.055 71.12 0.3 10 23.3293 7.018455 13.564 1.9549 19.212 15.954 58.0517PIPR83S 5.1028 85.07582 22 7.33 79.629 0.3 10 22.793 6.857111 13.794 1.9549 19.212 15.954 58.0517PIPR84S 5.4229 88.0785 22 7.605 99.53625 0.3 10 23.5975 7.099127 13.564 1.9549 19.212 15.954 59.0526PIPR85S 6.5439 87.0776 22 7.745 132.715 0.3 10 23.3293 7.018455 14.484 1.9549 18.731 15.954 59.0526PIPR86S 5.4310 87.0776 22 8.07 137.16 0.3 10 23.3293 7.018455 12.644 1.9549 18.731 15.954 59.0526PIPR87S 5.4306 87.0776 22 8.375 182.245 0.3 10 23.3293 7.018455 13.334 1.9549 18.731 15.954 59.0526PIPR88S 5.7955 87.0776 22 8.73 268.9225 0.3 10 23.3293 7.018455 14.254 1.9549 18.731 14.89 59.0526PIPR89S 6.9862 87.0776 22 8.115 188.976 0.3 10 23.3293 7.018455 12.874 1.9549 18.731 15.954 59.0526PIPR90S 8.5781 251.2239 22 7.2 662.559 0.3 10 67.127 20.35751 57.475 4.6918 72.524 62.397 150.133PIPR91S 9.0461 252.2248 22 7.575 904.875 0.3 10 67.3945 20.43861 57.475 4.6918 70.603 62.043 164.146PIPR92S 8.7054 252.2248 22 7.915 995.68 0.3 10 67.3945 20.43861 57.475 4.6918 73.484 62.043 150.133PIPR93S 6.4404 251.2239 22 8.275 891.54 0.3 10 67.127 20.35751 57.475 4.6918 73.484 62.043 143.127PIPR94S 8.4348 200.1784 22 8.05 757.6185 0.3 10 53.5426 16.18781 37.243 3.5188 49.47 46.798 128.114PIPR95S 8.0730 140.1249 22 7.95 404.8125 0.3 10 37.4414 11.35479 22.99 2.3459 28.817 25.172 99.0883PIPR96S 8.8271 90.08028 22 8.045 262.128 0.3 10 24.1338 7.260471 14.254 1.9549 18.731 15.599 65.0579PIPR97S 2.3840 19.01695 22 7.525 20.447 0.3 10 5.08133 1.541007 3.4485 0.782 0.9606 4.9634 19.0169PIPR98S 2.6680 34.03033 22 7.53 23.1648 0.3 10 9.0929 2.757591 3.4485 0.782 9.6058 4.6089 20.0178PIPR99S 4.5268 51.04549 22 7.54 34.9885 0.3 10 13.6394 4.136386 3.4485 0.782 16.81 4.6089 21.0187

PIPR100S 3.5167 29.02587 22 7.585 27.94 0.3 10 7.75571 2.352063 3.4485 0.782 5.2832 4.6089 22.0196PIPR101S 3.1703 30.02676 22 7.605 26.67 0.3 10 8.02315 2.433168 1.3794 0.782 4.3226 2.4817 23.0205PIPR102S 1.9033 27.02408 22 7.55 20.32 0.3 10 7.22084 2.189852 10.345 1.1729 5.2832 13.118 20.0178PIPR103S 2.9068 27.02408 22 7.525 26.67 0.3 10 7.22084 2.189852 20.691 1.5639 10.566 26.59 20.0178PIPR104S 6.9464 90.08028 22 7.995 182.88 0.3 10 24.1338 7.260471 14.254 1.9549 19.212 15.954 63.0561PIPR105S 4.3303 60.05352 22 8.11 96.6724 0.3 10 16.0463 4.866337 11.955 1.5639 3.8423 17.372 58.0517

E-9
Appendix E. BLM Table

Model Output Model Input

BLM

Data Label

Critical 

Accumulation

Hardness

(mg/L)

Temp

(oC) pH

Dissolved 

LC50 (μg/L)

DOC

(mg/L)

Humic 

Acid (%)

Ca

(mg/L)

Mg

(mg/L)

Na

(mg/L)

K

(mg/L)

SO4

(mg/L)

Cl 

(mg/L)

Alkalini

(mg/L

PIPR106S 6.1231 120.107 22 8.09 182.88 0.3 10 32.0926 9.732674 11.955 1.5639 33.62 17.372 59.0526PIPR107S 5.3380 180.1606 22 8.09 190.6905 0.3 10 48.1389 14.59901 11.955 1.5639 62.438 17.017 58.0517PIPR108S 4.7175 91.08117 22 8.125 127.0635 0.3 10 24.3369 7.380611 11.955 1.5639 19.212 15.954 59.0526PIPR109S 5.7327 90.08028 22 8.155 148.59 0.3 10 24.0695 7.299505 2.299 6.2557 15.85 6.027 60.0535PIPR110S 6.5363 93.08296 22 8.135 223.52 0.3 10 24.8718 7.542822 35.864 3.9098 27.377 49.989 62.0553PIPR111S 6.7795 92.08206 22 8.145 283.1465 0.3 10 24.6043 7.461717 71.728 7.4287 41.305 102.81 61.0544PIPR112S 5.0174 91.08117 22 8.19 150.241 0.3 10 24.402 7.341142 14.484 15.248 18.731 17.372 62.0553PIPR113S 6.2630 144.1284 22 8.38 644.525 0.3 10 38.5111 11.67921 34.485 3.1279 12.488 42.189 138.123PIPR114S 5.5141 292.2605 22 8.27 697.5475 0.3 10 78.092 23.68284 34.485 3.1279 87.893 57.079 137.122PIPR115S 5.1749 440.3925 22 8.225 752.475 0.3 10 117.673 35.68647 34.485 3.1279 175.31 41.125 133.118PIPR116S 5.8459 217.1936 22 8.31 653.415 0.3 10 58.0341 17.59992 34.485 3.1279 46.588 43.253 133.118PIPR117S 6.1591 218.1945 22 8.305 646.3665 0.3 10 58.3016 17.68102 6.8969 1.5639 38.903 9.5723 140.124PIPR118S 5.9250 212.1891 22 8.345 939.8 0.3 10 56.6969 17.19439 103.45 7.8197 65.319 124.79 143.127PIPR119S 8.2172 92.08206 22 8.125 253.365 0.3 10 24.6701 7.421814 14.254 1.9549 19.212 16.663 63.0561

PIPR120F 0.3052 48 25 8.03 109.44 2.64 10 14.1077 3.111984 1.35 0.57 3.54 1.25 44 PIPR121F 0.3617 45 25 8.04 116.16 2.64 10 13.2259 2.917485 1.27 0.57 3.33 1.17 44 PIPR122F 0.1755 46 25 7.98 84.96 2.64 10 13.5198 2.982318 1.3 0.57 3.4 1.2 41 PIPR123F 3.4889 30 25 6.82 418.56 10.4652 10 7.1362 2.964634 1.625 0.5 6.125 1.25 21 PIPR124F 1.8656 37 25 7.28 495.36 11.3373 10 8.80131 3.656382 2.0042 0.5 7.5542 1.5417 21 PIPR125F 2.8066 87 25 7.11 1522.56 31.3956 10 20.6978 8.4403 16.071 1.855 22.35 18.629 20 PIPR126F 3.1774 73 25 6.94 1083.84 24.4188 10 17.2174 7.3329 10.539 1.5232 18.439 13.619 18 PIPR127F 1.4538 84 25 7.07 528 14.5155 10 20.4644 8.008 6 1.4 34.5 10.95 12 PIPR128F 1.0075 66 25 6.97 960.96 32.9018 10 16.0792 6.292 4.7143 1.4 27.107 8.6036 12 PIPR129F 1.2809 43.9 25 7.4 88.32 2 10 12.9026 2.846168 1.24 0.57 3.24 1.14 42.4 PIPR130F 0.0860 47.04192 22 8.1 27.94 1.1 10 13.9359 2.983276 1.6093 0.391 3.362 1.4181 42.5379PIPR131F 1.1899 243.2168 22 8.01 105.7275 1.1 10 92.7261 2.884195 47.129 0.391 3.362 143.23 43.0383PIPR132F 0.1230 255.7279 22 8.01 40.0558 1.1 10 14.1661 53.5752 1.6093 0.391 3.362 143.23 43.5388PIPR133F 0.4522 47.04192 22 8.1 64.262 1.1 10 13.9359 2.983276 47.589 0.391 3.362 72.324 43.5388PIPR134F 0.3833 45.04014 22 8.02 49.01565 1.1 10 13.3428 2.856328 1.6093 0.391 3.362 1.4181 43.0383PIPR135F 0.3216 45.04014 22 8.65 67.7164 1.1 10 13.3428 2.856328 1.6093 0.391 3.362 1.4181 47.0419PIPR136F 0.1834 45.54059 22 7.3 18.669 1.1 10 13.4911 2.888065 1.6093 0.391 3.362 1.4181 44.0392PIPR137F 0.1256 49.04371 22 6.63 6.1468 1.1 10 14.5289 3.110224 1.6093 0.391 3.362 1.4181 49.0437PIPR138F 0.2961 45.04014 22 7.16 20.447 1.1 10 13.3428 2.856328 1.6093 0.391 3.362 15.599 26.0231PIPR139F 2.8408 43.03836 22 7.93 93.36405 1.1 10 12.7498 2.72938 1.6093 0.391 3.362 1.4181 41.0365

E-10
Appendix E. BLM Table

Model Output Model Input

BLM

Data Label

Critical 

Accumulation

Hardness

(mg/L)

Temp

(oC) pH

Dissolved 

LC50 (μg/L)

DOC

(mg/L)

Humic 

Acid (%)

Ca

(mg/L)

Mg

(mg/L)

Na

(mg/L)

K

(mg/L)

SO4

(mg/L)

Cl 

(mg/L)

Alkalini

(mg/L

PIPR140F 0.0373 45.54059 22 7.91 245.364 6.1 83.7705 13.4911 2.888065 1.6093 0.391 3.362 1.4181 44.0392PIPR141F 1.3667 45.04014 22 7.94 72.3392 1.1 10 13.3428 2.856328 1.6093 0.391 3.362 1.4181 43.0383PIPR142F 0.0310 45.04014 22 7.95 229.8065 6.1 83.7705 13.3428 2.856328 1.6093 0.391 3.362 1.4181 43.0383PIPR143F 0.1023 45.54059 22 7.94 195.453 3.6 72.5 13.4911 2.888065 1.6093 0.391 3.362 1.4181 44.0392PIPR144F 0.1038 45.04014 22 7.91 109.347 2.35 57.8723 13.3428 2.856328 1.6093 0.391 3.362 1.4181 42.0374PIPR145F 1.9076 44.03925 22 7.87 78.0034 1.1 10 13.0463 2.792854 1.6093 0.391 3.362 1.4181 42.0374PIPR146F 0.4905 44.03925 22 7.84 45.52315 1.1 10 13.0463 2.792854 1.6093 0.391 3.362 19.145 17.0151PIPR147F 1.3078 22.52007 22 6.01 4.3815 0.3 10 6.01736 1.824876 3.4485 0.391 3.362 4.2544 15.0133PIPR148F 1.5995 24.02141 22 7.02 12.4333 0.3 10 6.41852 1.946535 3.6784 0.391 3.362 4.9634 17.0151PIPR149F 2.4015 23.02052 22 8 26.8605 0.3 10 6.15108 1.865429 4.1382 0.782 3.362 4.9634 17.5156PIPR150F 2.3670 21.51918 22 9.01 51.3334 0.3 10 5.74992 1.743771 4.598 1.5639 3.362 4.9634 19.0169

PTLU01S 4.0390 173 22 8.3 364.8 0.5 10 28.5511 24.739 53.583 4.282 166.08 3.8824 117 PTLU02S 9.0637 173 22 7.25 460.8 0.5 10 28.5511 24.739 53.583 4.282 166.08 3.8824 117 

PTOR01F 0.2752 25 7.8 7.3 22.08 1.1 10 7.1535 1.9754 4.8154 0.7 3.997 5.9792 25 PTOR02F 0.1587 54 11.5 7.3 17.28 1.1 10 15.0937 3.6371 6.8831 0.7 12.163 9.6854 43 

XYTE01S 2.6511 173 22 8.15 211.2 0.5 10 28.5511 24.739 53.583 4.282 166.08 3.8824 117 XYTE02S 4.5011 173 22 8.05 326.4 0.5 10 28.5511 24.739 53.583 4.282 166.08 3.8824 117 

POAC01S 2.2126 167 22 8 153.6 0.5 10 27.5609 23.881 51.724 4.1335 160.32 3.7478 115 

LEMA01R 25.6628 85 20.2 7.3 2200 1.1 10 23.9 6.5 0.64 0.46 4.32 1.5 82 

LEMA02F 25.8381 45 20 7.5 1056 1.1 10 13.2259 2.917485 1.3 0.57 3.4 1.2 43 LEMA03F 27.6113 25.9 19 7.03 960 1.5 10 6.38814 2.42165 5.4743 1.6 26.489 19.425 27.1 LEMA04F 22.5658 85 21.85 7.45 1300 1.1 10 23.9 6.5 0.64 0.46 4.32 1.5 82 

ETFL01S 5.5744 170 20 7.8 316.8 0.5 10 27.9 24.2 52.5 4.2 163 3.80 115 ETFL02S 5.7421 170 20 7.8 327.36 0.5 10 27.9 24.2 52.5 4.2 163 3.80 115 ETFL03S 5.8278 170 20 7.9 358.08 0.5 10 27.9 24.2 52.5 4.2 163 3.80 115 ETFL04S 6.4920 170 20 7.8 376.32 0.5 10 27.9 24.2 52.5 4.2 163 3.80 115 

ETLE01S 3.7314 167 22 8 249.6 0.5 10 27.5609 23.881 51.724 4.1335 160.32 3.7478 115 

ETNI01S 7.8536 170 20 7.8 473.28 0.5 10 27.9 24.2 52.5 4.2 163 3.80 115 ETNI02S 7.7256 170 20 7.8 463.68 0.5 10 27.9 24.2 52.5 4.2 163 3.80 115 ETNI03S 9.1617 170 20 7.8 577.92 0.5 10 27.9 24.2 52.5 4.2 163 3.80 115 ETNI04S 8.5329 170 20 7.8 526.08 0.5 10 27.9 24.2 52.5 4.2 163 3.80 115 

ETRU01S 0.4735 167 22 8.2 57.6 0.5 10 27.5609 23.881 51.724 4.1335 160.32 3.7478 115 

BUBO01S 1.7185 167 22 7.9 115.2 0.5 10 27.5609 23.881 51.724 4.1335 160.32 3.7478 115 

E-11
Appendix F. Regression Plots
Species Study Test Endpoint

Control Value EC50 Standard Deviation 

Snail,Campeloma decisum (Test 1)

Arthur and Leonard 1970 LC Survival 0.925 14.50 0.192 

Snail,Campeloma decisum (Test 2)

Arthur and Leonard 1970 LC Survival 0.875 11.80 0.339 

Cladoceran,Daphnia pulex

Winner 1985 LC Survival 1.00 4.57 0.260 

Cladoceran,Daphnia pulex

Winner 1985 LC Survival 0.900 11.3 0.111 

Caddisfly,Clistoronia magnifica

Nebeker et al. 1984b LC Emergence (adult 

1st gen)

0.750 20.0 0.300 

Bluegill (larval),Lepomis macrochirus

Benoit 1975 ELS Survival 0.880 39.8 0.250 

Species Study Test Endpoint

Control Value EC50 Slope 

Cladoceran,Ceriodaphnia dubia

Carlson et al. 1986 LC Reproduction 13.10 14.6 1.36 

Cladoceran,Daphnia magna

Chapman et al. Manuscript LC Reproduction 171.5 16.6 1.40 

Cladoceran,Daphnia magna

Chapman et al. Manuscript LC Reproduction 192.1 28.4 1.59 

Cladoceran,Daphnia magna

Chapman et al. Manuscript LC Reproduction 88.0 15.8 1.00 

Rainbow trout,Oncorhynchus mykiss

Seim et al. 1984 ELS Biomass 137.6 40.7 1.69 

Rainbow trout,Oncorhynchus mykiss

Besser et al. 2001 ELS Biomass 1224 29.2 1.99 

Chinook salmon,Oncorhynchus tshawytscha

Chapman 1975, 1982 ELS Biomass 0.901 9.55 1.27 

Fathead minnow,Pimephales promelas

Lind et al. manuscript ELS Biomass 108.4 11.4 4.00 

Appendix F. Analyses of Chronic Data The following pages contain figures and other information related to the regression and probability distribution analyses that were pchronic EC20s. The initial parameter estimates are shown in the tables below. In the figures that follow, circles denote measured redenote estimated regression lines. 

Final EstimatesFinal Estimates

Probability Distribution Analysis 

Logistic Regression Analysis

F-1
F-2

Evaluation of the Chronic Data Available for Freshwater Species

Following is a species-by-species discussion of each chronic test on copper evaluated for this

document. Also presented are the results of regression analysis and probability distribution analysis of

each dataset that was from an acceptable chronic test and contained sufficient acceptable data. For each

such dataset, this appendix contains a figure that presents the data and regression/probability distribution

line.

Brachionus calyciflorus. The chronic toxicity of copper was ascertained in 4-day renewal tests

conducted at regular intervals throughout the life of the freshwater rotifer, B. calyciflorus (Janssen et al.

1994). The goal of this study was to develop and examine the use of this rotifer as a viable test organism.

The effect of copper on the age-specific survivorship and fertility of B. calyciflorus was determined, but

no individual replicate data were provided and only three copper concentrations were tested, which

precludes these data from further regression analysis. Chronic limits based on the intrinsic rate of natural

increase were 2.5 μg/L total copper (NOAEC) and 5.0 μg/L total copper (LOAEC). The chronic value

determined via traditional hypothesis testing is 3.54 μg/L total copper (Table 2a). 

Campeloma decisum. Adult C. campeloma were exposed to five concentrations of total copper

and a control (Lake Superior water) under flow-through conditions in two 6-week studies conducted by

Arthur and Leonard (1970). Adult survival in the two separate chronic copper toxicity test trials was

markedly reduced in the two highest copper concentrations, 14.8 and 28.0 μg/L, respectively. The

authors reported that growth, as determined from cast exoskeleton, was not measurable for this test

species, although the authors did observe that the adult snails would not consume food at the two highest

copper concentrations. Control survival was 80 percent or greater. Chronic values of 10.88 μg/L total

copper were obtained for survival based on the geometric mean of the NOAEC and LOAEC of 8.0 and

14.8 μg/L, respectively, in both tests. The corresponding EC20s were 8.73 and 10.94 μg/L (Table 2a). 

Ceriodaphnia dubia. The chronic toxicity of copper to C. dubia was determined in ambient river

water collected upstream of known point-source discharges of domestic and industrial wastes as part of a

water effect ratio study (Carlson et al. 1986). In this study, survival and young production of C. dubia

were assessed using a 7-day life-cycle test. Organisms were not affected at total copper concentrations

ranging from 3 to 12 μg/L (5 to 10 μg/L dissolved copper). There was a 62.7 percent reduction in

survival and 97 percent reduction in the mean number of young produced per female at 32 μg/L total

copper (27 μg/L dissolved copper). No daphnids survived to produce young at 91 μg/L total copper.

Control survival during the study was 80 percent, which included one male. The chronic value EC20

selected for C. dubia in this study, 9.17 μg/L derived from a nonlinear regression evaluation, was based

on mean number of young produced (reproduction).

The effects of water hardness on the chronic toxicity of copper to C. dubia were assessed by

Belanger et al. (1989) using 7-day life-cycle tests. C. dubia 2 to 8 hours old were exposed to copper in

ambient surface water from the New and Clinch Rivers, Virginia. Mean water hardness levels were 179

and 94 mg/L as CaCO3, respectively. Test water was renewed on days 3 and 5. The corresponding

chronic values for reproduction based on the NOAEC and LOAEC approach were 7.9 and <19.3 μg/L

dissolved copper, respectively. The EC20 value for number of young (neonates) produced in Clinch

River water (water hardness of 94 mg/L as CaCO3) was 19.36 μg/L dissolved copper. The EC20 for

young produced in New River water was not calculated. The chronic values were converted to total

copper using the freshwater conversion factor for copper 0.96 (e.g., 7.897/0.96). The resulting total

chronic values for the New and Clinch rivers are 8.23 and 20.17 μg/L, respectively. 
F-3

Copper was one of 12 toxicants examined by Oris et al. (1991) in their comparisons between a 4-

day survival and reproduction toxicity test utilizing C. dubia and a standard 7-day life-cycle test for the

species. The reported 7-day chronic values for survival and reproduction (mean total young per living

female) in two tests based on the traditional hypothesis testing techniques were 24.5 and 34.6 μg/L total

copper. Comparable point estimates for these 7-day tests could not be calculated using regression

analysis.

Daphnia magna. Blaylock et al. (1985) reported the average numbers of young produced for six

broods of D. magna in a 14-day chronic exposure to copper. A significant reduction was observed in the

mean number of young per female at a concentration of 30 μg/L total copper, the highest copper

concentration tested. At this concentration, young were not produced at brood intervals 5 and 6.

Reproduction was not affected at 10 μg/L total copper. The chronic value determined for this study

(17.32 μg/L total copper) was based on the geometric mean of the NOAEC, 10 μg/L, and LOAEC, 30

μg/L. 

Van Leeuwen et al. (1988) conducted a standard 21-day life-cycle test with D. magna. The water

hardness was 225 mg/L as CaCO3. Carapace length was significantly reduced at 36.8 μg/L total copper,

although survival was 100 percent at this concentration. Carapace length was not affected at 12.6 μg/L

total copper. No daphnids survived at 110 μg/L concentration. The highest concentration not

significantly different from the control for survival was 36.8 μg/L. The lowest concentration significantly

different from the control based on survival was 110 μg/L, resulting in a chronic value of 63.6 μg/L for

survival. The chronic value based on carapace length was 21.50 μg/L. The 21-day EC10 as reported by

the author was 5.9 μg/L total copper.

Chronic (21-day) renewal toxicity tests were conducted using D. magna to determine the

relationship between water hardness (nominal values of 50, 100, and 200 mg/L as CaCO3, respectively)

and the toxicity of total copper (Chapman et al. unpublished manuscript). All test daphnids were <1 day

old at the start of the tests. The dilution water was well water from the Western Fish Toxicology Station

(WFTS), Corvallis, Oregon. Test endpoints were reproduction (total and live young produced per female)

and adult survival. The survival of control animals was 100 percent at nominal water hardness levels of

50 and 200 mg/L as CaCO3

, and 80 percent at a hardness of 100 mg/L as CaCO3

. The chronic values for

total young produced per female (fecundity) based on the geometric mean of the NOAEC and LOAEC

were 13.63, 29.33, and 9.53 μg/L at the nominal hardness levels of 50, 100, and 200 mg/L as CaCO3

,

respectively. The corresponding EC20 values for reproduction calculated using nonlinear regression

analysis were 12.58, 19.89, and 6.06 μg/L total copper. The chronic toxicity of copper to D. magna was

somewhat ameliorated from an increase in water hardness from 50 to 100 mg/L as CaCO3

, but slightly

increased from 100 to 200 mg/L as CaCO3. 

Daphnia pulex. Winner (1985) evaluated the effects of water hardness and humic acid on the

chronic toxicity (42-day) of copper to D. pulex. Contrary to the expectation that sublethal endpoints are

more sensitive indicators of chronic toxicity, reproduction was not a sensitive indicator of copper stress

in this species. Water hardness also had little effect on the chronic toxicity of copper (similar to D.

magna trends), but humic acid significantly reduced chronic toxicity of copper when added to the varying

water types. The survival chronic values based on the NOAEC and LOAEC values for the three low to no

humic acid studies were 4.90, 7.07, and 12.25 μg/L total copper at hardnesses of 57.5, 115, and 230 (0.15

mg/L HA) μg/L as CaCO3, respectively. The EC20 values calculated for the low and high hardness

studies using nonlinear regression techniques were 2.83 and 9.16 μg/L at hardness values of 57.5 and 230

(0.15 mg/L HA) μg/L as CaCO3, respectively. 
F-4

Clistoronia magnifica. The effects of copper on the lifecycle of the caddisfly, C. magnifica, were

examined in Nebeker et al. (1984b). The test included continuous exposure of first-generation aquatic

larvae and pupae through to a third generation of larvae. A significant reduction in adult emergence

occurred at 13.0 μg/L total copper from first-generation larvae. No observed adverse effect to adult

emergence occurred at 8.3 μg/L total copper. Percent larval survival was close to the control value of 80

percent. The chronic value based on hypothesis testing was 10.39 μg/L total copper. The corresponding

EC20 value for adult emergence was 7.67 μg/L total copper. 

Oncorhynchus mykiss. The growth and survival of developing O. mykiss embryos continuously

and intermittently exposed to copper for up to 85 days post-fertilization was examined by Seim et al.

(1984). Results only from the continuous exposure study are considered here for deriving a chronic

value. A flow-through apparatus was used to deliver six concentrations and a control (untreated well

water; average of 3 μg/L copper) to a single incubation chamber. Continuous copper exposure of

steelhead embryos in the incubation chambers was begun 6 days post-fertilization. At 7 weeks postfertilization, when all control fish had hatched and reached swim-up stage, subsamples of approximately

100 alevins were transferred to aquaria and the same exposure pattern continued. Dissolved oxygen

remained near saturation throughout the study. Water hardness averaged 120 mg/L as CaCO3. Survival of

steelhead embryos and alevins exposed continuously to total copper concentrations in the range of 3

(controls) to 30 μg/L was greater than 90 percent or greater. Survival was reduced at 57 μg/L and

completely inhibited at 121 μg/L. A similar effect on survival was observed for embryos and alevins

exposed to a mean of 51 (peak 263) and 109 (peak 465) μg/L of copper in the intermittent exposure,

respectively. The adverse effect of continuous copper exposure on growth (measured on a dry weight

basis) was observed at concentrations as low as 30 μg/L. (There was a 30 percent reduction in growth

during the intermittent exposure at 16 μg/L.) The chronic limits for survival of embryos and alevin

steelhead trout exposed continuously to copper were 16 and 31 μg/L, respectively (geometric mean =

22.27 μg/L). The EC20 for biomass for the continuous exposure was 27.77 μg/L. 

Besser et al. (2001) conducted an ELS toxicity test with copper and the rainbow trout, O. mykiss,

starting with eyed embryos and continuing for 30 days after the fish reached the swim-up stage. The total

test period was 58 days. The test was conducted in ASTM moderately hard reconstituted water with a

hardness of approximately 160 to 180 mg/L as CaCO3

. Twenty-five eyed embryos were held in each of

four replicate egg cups at each concentration. Survival was monitored daily. At the end of the test,

surviving fish in each replicate chamber were weighed (dry weight). Dry weights were used to determine

growth and biomass of surviving fish. The no observed effect concentrations (NOECs) for survival and

biomass were both 12 μg/L and the lowest observed effect concentrations (LOECs) for survival and

biomass was also the same for both endpoints, 22 μg/L. The chronic values for biomass and survival

based on the geometric mean of the NOEC and LOEC were 16.25 μg/L. The corresponding EC20 for

biomass was 20.32 μg/L.

Oncorhynchus tshawytscha. The draft manuscript prepared by Chapman (1975/1982) provides

the results from a 4-month egg through fry partial chronic test conducted to determine the effects of

copper on survival and growth of O. tshawytscha. Continuous exposure occurred from several hours

post-fertilization through hatch, swim-up, and feeding fry stages. The test was terminated after 14 weeks

post-hatch. The dilution water was WFTS well water. Because of the influence of the nearby Willamette

River on the hardness of this well water, reverse osmosis water was mixed periodically with ambient well

water to attain a consistent hardness. The typical hardness of this well water was approximately 23 mg/L

as CaCO3

. Control survival exceeded 90 percent for the test. The measured total copper concentrations

during the test were 1.2 (control), 7.4, 9.4, 11.7, 15.5, and 20.2 μg/L, respectively. Copper adversely

affected survival at 11.7 μg/L copper and higher, and growth was reduced at all copper concentrations

tested compared with the growth of control fish. The chronic limits for copper in this study were
F-5

estimated to be less than 7.4 μg/L. The EC20 value estimated for biomass is 5.92 μg/L total copper based

on a logistic nonlinear regression model. 

Salmo trutta. McKim et al. (1978) examined the survival and growth (expressed as standing

crop) of embryo-larval and early juvenile brown trout to copper. The most sensitive exposure was with

embryos exposed for 72 days. The NOAEC and LOAEC, as obtained from the figure, were 20.8 and 43.8

μg/L total copper, respectively. Data were not available to calculate point estimates at the 20 percent

effect level using regression analysis. The chronic value selected for this species was 29.91 μg/L total

copper (geometric mean of 20.8 and 43.8 μg/L total copper). 

Salvelinus fontinalis. Sauter et al. (1976) examined the effects of copper on selected freshwater

fish species at different hardness levels (softwater at 37.5 mg/L as CaCO3; hardwater at 187 mg/L as

CaCO3

) during a series of partial life-cycle (PLC) tests. The species tested were brook trout (Salvelinus

fontinalis), channel catfish (Ictalurus punctatus), and walleye (Stizostedion vitreum). Because of the poor

embryo and larval survival of control animals (in all cases less than 70 percent), results from tests with

channel catfish and walleye were not included in Table 2a. One of the replicate control chambers from

the PLC tests conducted with brook trout in hard water also exhibited poor hatchability (48 percent) and

survival (58 percent) between 31 and 60 days of exposure. Therefore, the data for brook trout in hard

water were not included in the subsequent EC20 (regression) analysis either. 

The softwater test with brook trout was conducted using untreated well water with an average

water hardness of 35 mg/L as CaCO3

. This PLC exposure consisted of six copper concentrations and a

control. Hatchability was determined by examining randomly selected groups of 100 eggs from each

replicate exposure tank. Growth and survival of fry were determined by impartially reducing the total

sample size to 50 fry per tank and assessing their progress over 30 day intervals up to 60 days post-hatch.

The chronic limits based on the growth (wet weight and total length) of larval brook trout after 60 days of

exposure to copper in soft water were <5 and 5 μg/L. The resultant chronic value for soft water based on

hypothesis testing was <5 μg/L. The corresponding EC20 values based on total length, wet weight, and

biomass (the product of wet weight and survival) for brook trout in the soft-water exposures after 60 days

were not amenable to nonlinear regression analysis. 

McKim et al. (1978) examined survival and growth (expressed as standing crop) of embryolarval and early juvenile brook trout exposed to copper. The embryo exposure was for 16 days, and the

larval-early-juveniles exposure lasted 60 days. The NOAEC and LOAEC were 22.3 and 43.5 μg/L total

copper, respectively. Data were not available to calculate point estimates at the 20 percent effect level

using regression analysis. The chronic value for this species was 31.15 μg/L total copper (geometric

mean of 22.3 and 43.5 μg/L total copper).

Salvelinus namaycush. McKim et al. (1978) examined the survival and growth (expressed as

standing crop) of embryo-larval and early juvenile lake trout exposed to copper. The embryo exposure

was for 27 days, and the larval-early-juveniles exposure lasted 66 days. The NOAEC and LOAEC were

22.0 and 43.5 μg/L total copper, respectively. Data were not available to calculate point estimates at the

20 percent effect level using regression analysis. The chronic value for this species was 30.94 μg/L total

copper (geometric mean of 22.0 and 43.5 μg/L total copper). 

Esox lucius. McKim et al. (1978) examined the survival and growth (expressed as standing crop)

of embryo-larval and early juvenile northern pike exposed to copper. The embryo exposure was for 6

days, and the larval-early-juveniles exposure lasted 34 days. The NOAEC and LOAEC were 34.9 and

104.4 μg/L total copper, respectively. The authors attributed the higher tolerance of E. lucius to copper to

the very short embryonic exposure period compared with salmonids and white sucker, Catostomus
F-6

commersoni. Data were not available to calculate point estimates at the 20 percent effect level using

regression analysis. The chronic value for this species was 60.36 μg/L total copper (geometric mean of

34.9 and 104.4 μg/L total copper). 

Pimephales notatus. An experimental design similar to that described by Mount and Stephan

(1967) and Mount (1968) was used to examine the chronic effect of copper on the bluntnose minnow, P.

notatus (Horning and Neiheisel 1979). Measured total copper concentrations were 4.3 (control), 18.0,

29.9, 44.1, 71.8, and 119.4 μg/L, respectively. The experimental dilution water was a mixture of spring

water and demineralized City of Cincinnati tap water. Dissolved oxygen was kept at 5.9 mg/L or greater

throughout the test. Total water hardness ranged from 172 to 230 mg/L as CaCO3. The test was initiated

with 22 6-week-old fry. The fish were later separated according to sex and thinned to a sex ratio of 5

males and 10 females per duplicated test chamber. Growth (total length) was significantly reduced in

parental and first (F1

) generation P. notatus after 60 days of exposure to the highest concentration of

copper tested (119.4 μg/L). Survival of parental P. notatus exposed to this same high test concentration

was also lower (87 percent) at the end of the test compared with the other concentrations (range of 93 to

100 percent). Copper at concentrations of 18 μg/L and greater significantly reduced the number of eggs

produced per female. The number of females available to reproduce was generally the same up to about

29.9 μg/L of copper. The chronic limits were based on an NOAEC and LOAEC of <18 and 18 μg/L for

number of eggs produced per female. An EC20 was not estimated by nonlinear regression; nevertheless,

in this case an EC20 is likely to be substantially below 18 :g/L.

Pimephales promelas. The results from a 30-day ELS toxicity test to determine the chronic

toxicity of copper to P. promelas using dilution water from Lake Superior (hardness ranging from 40 to

50 mg/L as CaCO3) was included in Table 2a from a manuscript prepared by Lind et al. in 1978. In this

experiment, five test concentrations and a control were supplied by a continuous-flow diluter. The

exposure began with embryos 1 day post-fertilization. Pooled results from fish dosed in replicate

exposure chambers were given for mean percentage embryo survival to hatch, mean percentage fish

survival after hatch, and mean fish wet weight after 30 days. The percentage of embryo survival to hatch

was not affected by total copper concentrations as high as 52.1 μg/L total copper. Survival after hatch,

however, was compromised at 26.2 μg/L, and mean wet weight of juvenile fathead minnows was

significantly reduced at 13.1 μg/L of copper. The estimated EC20 value for biomass was 9.376 μg/L total

copper. 

Catastomus commersoni. McKim et al. (1978) examined the survival and growth (expressed as

standing crop) of embryo-larval and early juvenile white sucker exposed to copper. The embryo exposure

was for 13 days, and the larval-early-juvenile exposure lasted 27 days. The NOAEC and LOAEC were

12.9 and 33.8 μg/L total copper, respectively. The resulting chronic value based on hypothesis testing for

this species was 20.88 μg/L total copper (geometric mean of 12.9 and 33.8 μg/L total copper).

Lepomis macrochirus. Results from a 22-month copper life-cycle toxicity test with bluegill (L.

macrochirus) were reported by Benoit (1975). The study included a 90-day embryo-larval survival and

growth component. The tests were conducted at the U.S. EPA National Water Quality Laboratory in

Duluth, Minnesota, using Lake Superior water as the dilution water (average water hardness = 45 mg/L

as CaCO3). The test was initiated in December 1969 with 2-year-old juvenile L. macrochirus. In May

1971, the fish were sexed and randomly reduced to three males and seven females per tank. Spawning

commenced on 10 June 1971. The 90-day embryo-larval exposure was initiated when 12 lots of 50 newly

hatched larvae from one of the two control groups were randomly selected and transferred to duplicate

grow-out chambers at 1 of 6 total copper concentrations: 3 (control), 12, 21, 40, 77, and 162 μg/L,

respectively. In the 22-month juvenile through adult exposure, survival, growth, and reproduction were

unaffected at 77 μg/L of copper and below. No spawning occurred at 162 μg/L. Embryo hatchability and
F-7

survival of 4-day-old larvae at 77 μg/L did not differ significantly from those of controls. However, after

90 days of exposure, survival of larval L. macrochirus at 40 and 77 μg/L was significantly lower than for

controls, and no larvae survived at 162 μg/L. Growth remained unaffected at 77 μg/L. Based on the 90-

day survival of bluegill larvae, the chronic limits were estimated to be 21 and 40 μg/L (geometric mean =

28.98 μg/L). The corresponding EC20 for embryo-larval survival was 27.15 μg/L. 
F-8

Campeloma decisum (Test 1), Life-cycle, Arthur and Leonard 1970

Campeloma decisum (Test 2), Life-cycle, Arthur and Leonard 1970

Ceriodaphnia dubia (Clinch River), Life-cycle, Belanger et al. 1989

EC20 = 8.73 μg/L

EC20 = 10.94 μg/L

EC20 = 19.36 μg/L
F-9

Lepomis macrochirus, Early Life-stage, Benoit 1975

Oncorhynchus mykiss, Early Life-Stage, Besser et al. 2001

Ceriodaphnia dubia, Life-cycle, Carlson et al. 1986

EC20 = 27.15 μg/L

EC20 = 20.32 μg/L

EC20 = 9.17 μg/L
F-10

Daphnia magna (Hardness 104), Life-cycle, Chapman et al. Manuscript

Daphnia magna (Hardness 211), Life-cycle, Chapman et al. Manuscript

Daphnia magna (Hardness 51), Life-cycle, Chapman et al. Manuscript

EC20 = 19.89 μg/L

EC20 = 6.06 μg/L

EC20 = 12.58 μg/L
F-11

EC20 = 7.67 μg/L

Oncorhynchus tshawytscha, Early Life-Stage, Chapman 1975 & 1982

Pimephales promelas, Early Life-stage, Lind et al. 1978

Clistoronia magnifica, Life-cycle, Nebeker et al. 1984a

EC20 = 5.92 μg/L

EC20 = 9.38 μg/L
F-12

Oncorhynchus mykiss, Early Life-stage, Seim et al. 1984

Daphnia pulex (Hardness 230 HA 0.15), Life-cycle, Winner 1985

Daphnia pulex (Hardness 57), Life-cycle, Winner 1985

EC20 = 27.77 μg/L

EC20 = 9.16 μg/L

EC20 = 2.83 μg/L
Appendix G. Example Water Quality Criteria Values Using the BLM and the

Hardness Equation
Appendix G: Representative water quality criteria values using the BLM and the 

Hardness equation approaches for waters with a range in pH, Hardness, and DOC

concentrations. The BLM calculation assumed that alkalinity was correlated with pH, and

that other major ions were correlated with hardness based on observed correlations in 

EPA synthetic water recipes.

pH Hardness DOC

Hardness 

Equation Based 

Water Quality 

Criterion for 

Cu[1]

BLM Based 

Instantaneous 

Water Quality 

Criterion for Cu

mg/L CaCO3 mg / L μg / L μg / L

2 5.9 1.6

4 5.9 3.3

8 5.9 6.8

16 5.9 14.3

2 11.3 1.9

4 11.3 3.8

8 11.3 7.7

16 11.3 16.0

2 21.7 2.3

4 21.7 4.5

8 21.7 9.2

16 21.7 18.9

2 41.5 2.8

4 41.5 5.6

8 41.5 11.4

16 41.5 23.1

2 5.9 3.9

4 5.9 8.0

8 5.9 16.4

16 5.9 34.3

2 11.3 4.4

4 11.3 8.8

8 11.3 18.0

16 11.3 37.0

2 21.7 5.1

4 21.7 10.3

8 21.7 20.7

16 21.7 42.4

2 41.5 6.2

4 41.5 12.4

8 41.5 24.9

16 41.5 50.6

6.5 40

80

159

317

7.0 40

80

159

317

G-1
pH Hardness DOC

Hardness 

Equation Based 

Water Quality 

Criterion for 

Cu[1]

BLM Based 

Instantaneous 

Water Quality 

Criterion for Cu

mg/L CaCO3 mg / L μg / L μg / L

2 5.9 7.9

4 5.9 15.8

8 5.9 32.4

16 5.9 67.3

2 11.3 8.7

4 11.3 17.4

8 11.3 35.3

16 11.3 72.5

2 21.7 10.1

4 21.7 20.1

8 21.7 40.5

16 21.7 82.4

2 41.5 12.0

4 41.5 23.9

8 41.5 47.8

16 41.5 96.8

2 5.9 13.8

4 5.9 27.6

8 5.9 55.8

16 5.9 115.0

2 11.3 15.5

4 11.3 30.6

8 11.3 61.4

16 11.3 125.1

2 21.7 18.0

4 21.7 35.3

8 21.7 70.3

16 21.7 142.0

2 41.5 21.5

4 41.5 41.6

8 41.5 82.3

16 41.5 165.1

7.5 40

80

159

317

8.0 40

80

159

317

G-2
pH Hardness DOC

Hardness 

Equation Based 

Water Quality 

Criterion for 

Cu[1]

BLM Based 

Instantaneous 

Water Quality 

Criterion for Cu

mg/L CaCO3 mg / L μg / L μg / L

2 5.9 22.5

4 5.9 43.3

8 5.9 85.6

16 5.9 172.9

2 11.3 26.0

4 11.3 49.1

8 11.3 96.0

16 11.3 191.6

2 21.7 31.4

4 21.7 58.0

8 21.7 111.7

16 21.7 220.6

2 41.5 39.1

4 41.5 70.3

8 41.5 132.8

16 41.5 259.6

Notes:

[1] : Hardness Equation: CMC = e (0.9422 [ln(H)] - 1.7)

where:

H = water hardness (mg/L CaCO3)

8.5 40

80

159

317

G-3
Appendix H. Unused Data
H-1

APPENDIX H. UNUSED DATA

Based on the requirements set forth in the guidelines (Stephan et al. 1985), the following studies

are not acceptable for the following reasons and are classified as unused data. 

Studies Were Conducted with Species That Are Not Resident in North America

Abalde et al. (1995)

Abel (1980)

Ahsanullah and Ying (1995)

Ahsanullah et al. (1981)

Aoyama and Okamura (1984)

Austen and McEvoy (1997)

Bougis (1965)

Cid et al. (1995, 1996a,b)

Collvin (1984)

Cosson and Martin (1981)

Daly et al. (1990a,b, 1992)

Denton and Burdon-Jones (1986)

Drbal et al. (1985)

Giudici and Migliore (1988)

Giudici et al. (1987, 1988)

Gopal and Devi (1991)

Gustavson and Wangberg (1995)

Hameed and Raj (1989)

Heslinga (1976)

Hori et al. (1996)

Huebner and Pynnonen (1992)

Ismail et al. (1990)

Jana and Bandyopadhyaya (1987)

Jindal and Verma (1989)

Jones (1997)

Kadioglu and Ozbay (1995)

Karbe (1972)

Knauer et al. (1997)

Kulkarni (1983)

Kumar et al. (1985)

Lan and Chen (1991)

Lee and Xu (1984)

Luderitz and Nicklisch (1989)

Majori and Petronio (1973)

Masuda and Boyd (1993)

Mathew and Fernandez (1992)

Maund et al. (1992)

Migliore and Giudici (1988)

Mishra and Srivastava (1980)

Negilski et al. (1981)

Nell and Chvojka (1992)

Neuhoff (1983)

Nias et al. (1993)

Nonnotte et al. (1993)

Pant et al. (1980)

Paulij et al. (1990)

Peterson et al. (1996)

Pistocchi et al. (1997)

Pynnonen (1995)

Raj and Hameed (1991)

Rajkumar and Das (1991)

Reeve et al. (1977)

Ruiz et al. (1994, 1996)

Saward et al. (1975)

Schafer et al. (1993)

Smith et al. (1993)

Solbe and Cooper (1976)

Steeman-Nielsen and Bruun-Laursen

(1976)

Stephenson (1983)

Takamura et al. (1989)

Taylor et al. (1991, 1994)

Timmermans (1992)

Timmermans et al. (1992)

Vardia et al. (1988)

Verriopoulos and MoraitouApostolopoulou (1982)

Visviki and Rachlin (1991)

Weeks and Rainbow (1991)

White and Rainbow (1982)

Wong and Chang (1991)

Wong et al. (1993)

Copper Was a Component of a Drilling Mud, Effluent, Mixture, Sediment, or Sludge

Buckler et al. (1987)

Buckley (1994)

Clements et al. (1988)

de March (1988)

Hollis et al. (1996)

Horne and Dunson (1995)

Hutchinson and Sprague (1987)

Kraak et al. (1993 and 1994a,b)

Lowe (1988)

McNaught (1989)

Munkittrick and Dixon (1987)

Pellegrini et al. (1993)

Roch and McCarter (1984a,b)

Roch et al. (1986)

Sayer et al. (1991b)

Weis and Weis (1993)

Widdows and Johnson (1988)

Wong et al. (1982)
H-2

These Reviews Only Contain Data That Have Been Published Elsewhere

Ankley et al. (1993)

Borgmann and Ralph (1984)

Chapman et al. (1968)

Chen et al. (1997)

Christensen et al. (1983)

Dierickx and Brendael-Rozen (1996)

DiToro et al. (1991)

Eisler (1981)

Eisler et al. (1979)

Enserink et al. (1991)

Felts and Heath (1984)

Gledhill et al. (1997)

Handy (1996)

Hickey et al. (1991)

Janssen et al. (1994)

LeBlanc (1984)

Lilius et al. (1994)

Meyer et al. (1987)

Ozoh (1992c)

Peterson et al. (1996)

Phillips and Russo (1978)

Phipps et al. (1995)

Spear and Pierce (1979b)

Starodub et al. (1987b)

Taylor et al. (1996)

Thompson et al. (1972)

Toussaint et al. (1995)

No Interpretable Concentration, Time, Response Data, or Examined Only a Single Concentration

Asztalos et al. (1990)

Beaumont et al. (1995a,b)

Beckman and Zaugg (1988)

Bjerselius et al. (1993)

Carballo et al. (1995)

Daoust et al. (1984)

De Boeck et al. (1995b, 1997)

Dick and Dixon (1985)

Felts and Heath (1984)

Ferreira (1978)

Ferreira et al. (1979)

Hansen et al. (1993, 1996)

Heath (1987, 1991)

Hughes and Nemcsok (1988)

Julliard et al. (1996)

Koltes (1985)

Kosalwat and Knight (1987)

Kuwabara (1986)

Lauren and McDonald (1985)

Leland (1983)

Lett et al. (1976)

Miller and McKay (1982)

Mis and Bigaj (1997)

Nalewajko et al. (1997)

Nemcsok et al. (1991)

Ozoh (1990)

Ozoh and Jacobson (1979)

Parrott and Sprague (1993)

Pyatt and Dodd (1986)

Riches et al. (1996)

Sayer (1991)

Sayer et al. (1991a,b)

Schleuter et al. (1995, 1997)

Starcevic and Zielinski (1997)

Steele (1989)

Taylor and Wilson (1994)

Viale and Calamari (1984)

Visviki and Rachlin (1994b)

Waiwood (1980)

Webster and Gadd (1996)

Wilson and Taylor (1993a,b)

Winberg et al. (1992)

Wundram et al. (1996)

Wurts and Perschbacher (1994)

No Useable Data on Copper Toxicity or Bioconcentration

Cowgill et al. (1986)

de March (1979)

Lehman and Mills (1994)

Lustigman (1986)

Lustigman et al. (1985)

MacFarlane et al. (1986)

van Hoof et al. (1994)

Weeks and Rainbow (1992)

Wong et al. (1977)

Wren and McCarroll (1990)

Zamuda et al. (1985)

Results Not Interpretable as Total or Dissolved Copper

Brand et al. (1986)

MacFie et al. (1994)

Riedel (1983)

Sanders and Jenkins (1984)

Sanders and Martin (1994)

Sanders et al. (1995)

Stearns and Sharp (1994)

Stoecker et al. (1986)

Sunda et al. (1987)

Winberg et al. (1992)

Some of these studies would be valuable if copper criteria were developed on the basis of cupric

ion activity.
H-3

Organisms Were Selected, Adapted or Acclimated for Increased Resistance to Copper

Fisher (1981)

Fisher and Fabris (1982)

Hall (1980)

Hall et al. (1989)

Harrison and Lam (1983)

Harrison et al. (1983)

Lumoa et al. (1983)

Lumsden and Florence (1983)

Munkittrick and Dixon (1989)

Myint and Tyler (1982)

Neuhoff (1983)

Parker (1984)

Phelps et al. (1983)

Ray et al. (1981)

Sander (1982)

Scarfe et al. (1982)

Schmidt (1978a,b)

Sheffrin et al. (1984)

Steele (1983b)

Takamura et al. (1989)

Viarengo et al. (1981a,b)

Wood (1983)

Either the Materials, Methods, Measurements or Results Were Insufficiently Described

Abbe (1982)

Alam and Maughan (1995)

Balasubrahmanyam et al. (1987)

Baudouin and Scoppa (1974)

Belanager et al. (1991)

Benedeczky et al. (1991)

Benedetti et al. (1989)

Benhra et al. (1997)

Bouquegneau and Martoja (1982)

Burton and Stemmer (1990)

Burton et al. (1992)

Cabejszek and Stasiak (1960)

Cain and Luoma (1990)

Chapman (1975, 1982)

Cochrane et al. (1991)

Devi et al. (1991)

Dirilgen and Inel (1994)

Dodge and Theis (1979)

Doucet and Maly (1990)

Dunbar et al. (1993)

Durkina and Evtushenko (1991)

Enesco et al. (1989)

Erickson et al. (1997)

Evans (1980)

Ferrando and Andreu (1993)

Finlayson and Ashuckian (1979)

Furmanska (1979)

Gibbs et al. (1981)

Gordon et al. (1980)

Gould et al. (1986)

Govindarajan et al. (1993)

Hayes et al. (1996)

Howard and Brown (1983)

Janssen et al. (1993)

Janssen and Persoone (1993)

Kean et al. (1985)

Kentouri et al. (1993)

Kessler (1986)

Khangarot et al. (1987)

Kobayashi (1996)

Kulkarni (1983)

Labat et al. (1977)

Lakatos et al. (1993)

LeBlanc (1985)

Leland et al. (1988)

Mackey (1983)

Magni (1994)

Martin et al. (1984)

Martincic et al. (1984)

McIntosh and Kevern (1974)

McKnight (1980)

Moore and Winner (1989)

Muramoto (1980, 1982)

Nyholm and Damgaard (1990)

Peterson et al. (1996)

Pophan and D’Auria (1981)

Reed-Judkins et al. (1997)

Rehwoldt et al. (1973)

Riches et al. (1996)

Sakaguchi et al. (1977)

Sanders et al. (1995)

Sayer (1991)

Schultheis et al. (1997)

See et al. (1974)

Shcherban (1977)

Smith et al. (1981)

Sorvari and Sillanpaa (1996)

Stearns and Sharp (1994)

Strong and Luoma (1981)

Sullivan and Ritacco (1988)

Taylor (1978)

Taylor et al. (1994)

Thompson (1997)

Trucco et al. (1991)

Verma et al. (1980)

Visviki and Rachlin (1994a)

Watling (1983)

Winner et al. (1990)

Young and Harvey (1988, 1989)

Zhokhov (1986)
H-4

Questionable Effect Levels Due to Graphical Presentation of Results

Alliot and Frenet-Piron (1990)

Andrew (1976)

Arsenault et al. (1993)

Balasubrahmanyam et al. (1987)

Bjerselius et al. (1993)

Bodar et al. (1989)

Chen (1994)

Cowgill and Milazzo (1991b)

Cvetkovic et al. (1991)

Dodoo et al. (1992)

Francisco et al. (1996)

Gupta et al. (1985)

Hansen et al. (1996)

Hoare and Davenport (1994)

Lauren and McDonald (1985)

Llanten and Greppin (1993)

Metaxas and Lewis (1991)

Michnowicz and Weeks (1984)

Miersch et al. (1997)

Nasu et al. (1988)

Pearlmutter and Lembi (1986)

Pekkala and Koopman (1987)

Peterson et al. (1984)

Romanenko and Yevtushenko (1985)

Sanders et al. (1994)

Smith and Heath (1979)

Stokes and Hutchinson (1976)

Winner and Gauss (1986)

Wong (1989)

Young and Lisk (1972)

Studies of Copper Complexation With No Useable Toxicology Data for Surface Waters

Borgmann (1981)

Filbin and Hough (1979)

Frey et al. (1978)

Gillespie and Vaccaro (1978)

Guy and Kean (1980)

Jennett et al. (1982)

Maloney and Palmer (1956)

Nakajima et al. (1979)

Stauber and Florence (1987)

Sunda and Lewis (1978)

Swallow et al. (1978)

van den Berg et al. (1979)

Wagemann and Barica (1979)

Questionable Treatment of Test Organisms or Inappropriate Test Conditions or Methodology

Arambasic et al. (1995)

Benhra et al. (1997)

Billard and Roubaud (1985)

Bitton et al. (1995)

Brand et al. (1986)

Bringmann and Kuhn (1982)

Brkovic-Popovic and Popovic 

(1977a,b)

Dirilgen and Inel (1994)

Folsom et al. (1986)

Foster et al. (1994)

Gavis et al. (1981)

Guanzon et al. (1994)

Hawkins and Griffith (1982)

Ho and Zubkoff (1982)

Hockett and Mount (1996)

Huebert et al. (1993)

Huilsom (1983)

Jezierska and Slominska (1997)

Kapu and Schaeffer (1991)

Kessler (1986)

Khangarot and Ray (1987a)

Khangarot et al. (1987)

Lee and Xu (1984)

Marek et al. (1991)

McLeese (1974)

Mis et al. (1995)

Moore and Winner (1989)

Nasu et al. (1988)

Ozoh and Jones (1990b)

Reed and Moffat (1983)

Rueter et al. (1981)

Sayer et al. (1989)

Schenck (1984)

Shaner and Knight (1985)

Sullivan et al. (1983)

Tomasik et al. (1995)

Watling (1981, 1982, 1983)

Wikfors and Ukeles (1982)

Wilson (1972)

Wong and Chang (1991)

Wong (1992)

High control mortalities occurred in all except one test reported by Sauter et al. (1976). Control

mortality exceeded 10% in one test by Mount and Norberg (1984). Pilgaard et al. (1994) studied

interactions of copper and hypoxia, but failed to run a hypoxic control. Beaumont et al. (1995a,b) studied

interactions of temperature, acid pH and copper, but never separated pH and copper effects. The 96-hour

values reported by Buikema et al. (1974a,b) were subject to error because of possible reproductive

interactions (Buikema et al. 1977).
H-5

Bioconcentration Studies Not Conducted Long Enough, Not Steady-State,

Not Flow-through, or Water Concentrations Not Adequately Characterized or Measured

Anderson and Spear (1980a)

Felton et al. (1994)

Griffin et al. (1997)

Harrison et al. (1988)

Krantzberg (1989)

Martincic et al. (1992)

McConnell and Harrel (1995)

Miller et al. (1992)

Ozoh (1994)

Wright and Zamuda (1987)

Xiaorong et al. (1997)

Yan et al. (1989)

Young and Harvey (1988, 1989)

Zia and Alikhan (1989)

Anderson (1994), Anderson et al. (1994), Viarengo et al. (1993), and Zaroogian et al. (1992)

reported on in vitro exposure effects. Benedeczky et al. (1991) studied only effects of injected copper.

Ferrando et al. (1993b) studied population effects of copper and cladoceran predator on the rotifer prey,

but the data are difficult to interpret. A similar problem complicated use of the cladoceran competition

study of LeBlanc (1985).