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OFFICE OF

CHEMICAL SAFETY AND

POLLUTION PREVENTION

UNITED STATES ENVIRONMENTAL PROTECTION AGENCY

WASHINGTON D.C., 20460

PC Codes: 110301 &

110302

DP Barcode: D425778 

Date: August 27, 2015

MEMORANDUM

SUBJECT: Problem Formulation and Preliminary Risk Assessment for Support of the 

Registration Review for Aquatic Dyes Containing Acid Blue 9 (PC Code 

110301) and Acid Yellow 23 (PC Code 110302)

FROM: Nicholas Mastrota, Ph.D., Wildlife Biologist

James K. Wolf, Ph.D., Environmental Scientist

Environmental Risk Branch 3

Environmental Fate and Effects Division (7507P)

REVIEWED Rosanna Louie-Juzwiak, Risk Assessment Process Leader

BY: Melissa Panger, Ph.D., Senior Biologist

James Hetrick, Ph.D., Senior Advisor

Environmental Risk Branch 3

Environmental Fate and Effects Division (7507P)

THROUGH: Dana Spatz, Branch Chief

Environmental Risk Branch 3

Environmental Fate and Effects Division (7507P)

TO: Neil Anderson, Branch Chief

Christina Motilall, Chemical Review Manager

Risk Management and Implementation Branch 1

Pesticide Re-Evaluation Division (7508P)

The Environmental Fate and Effects Division (EFED) has completed the preliminary ecological 

risk and drinking water assessments in support of the registration review of the aquatic dyes 

containing acid blue 9 (PC Code 110301) and acid yellow 23 (PC Code 110302). Alternative 

names for acid blue 9 and acid yellow 23 are erioglaucine and tartrazine, respectively. These dye 

mixtures are marketed under the trade names Aquashade® (EPA Reg. # 33068-1, 33068-2, 33068-

3) and Admiral® (EPA Reg # 67064-1, 67064-2). Since uses of these dyes have not changed, and 

no new environmental fate or ecotoxicity testing has been submitted, little has changed since 

these dyes were previously assessed for reregistration in 2005. Therefore, this abbreviated risk 

assessment provides updates to exposure and risk conclusions not previously assessed (i.e., 

chronic risk to mammals). This document serves as both the problem formulation and the 
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preliminary risk assessment in support of the registration review of aquatic dyes containing Acid 

Blue 9 and Acid Yellow 23.
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1. Executive summary

The Environmental Fate and Effects Division (EFED) has completed the preliminary ecological 

risk and drinking water assessments in support of the registration review of the aquatic dyes 

containing Acid Blue 9 (PC Code 110301) and Acid Yellow 23 (PC Code 110302). No new 

environmental fate or ecotoxicity testing has been submitted since the ecological risk assessment 

was conducted in 2005 for reregistration of these dyes. Also, no new uses or products have been 

added since that time. The restriction of these dyes to water bodies “under complete control of the 

user and have little or no outflow” and the prohibition of use in water that may be used for human 

consumption greatly limits exposure to drinking water resources or aquatic habitats other than 

those treated directly. Exposure to drinking water resources from registered use of these dyes is

also believed to be negligible. The available ecotoxicity data, together with information on risks 

to aquatic organisms available from the open literature, are sufficient to assess ecological risks.

Furthermore, all available toxicity information indicates that these dyes have very low toxicity 

and pose little risk to animals and plants at environmentally relevant concentrations.

2. Previous Assessments

EFED’s most recent ecological risk assessment for dyes containing Acid Blue 9 and Acid Yellow 

23 was completed for the previous reregistration process and was summarized in a Reregistration 

Eligibility Decision (RED1) from September 26, 2005 (D302183). Since that memorandum was 

completed, the only new information that EFED has obtained that is relevant to the ecological risk 

of these dyes has come from open literature papers. Several papers have been published on the 

toxicity of these dyes to aquatic organisms (or lack thereof), and their effects to fish, plankton, 

and aquatic plants when used in aquaculture systems. Therefore, much of this assessment consists 

of a review and summary of findings from these published papers, and a discussion of how this 

new information effects the risk conclusions that EFED had previously reported for use of these 

dyes.

3. Nature of Chemical Stressor

The chemical stressors are mixtures of acid blue 9 (also known as erioglaucine) and acid yellow 

23 (also known as tartrazine) that are used to color small water bodies blue-green to attenuate 

light penetration, and thereby reduce growth of nuisance aquatic plants and algae. While the 

proportions of the two dyes vary somewhat among the four registered products, acid blue 9 is 

always by far the larger component, comprising 92% to 94% of the total dye by weight. Ratios of 

acid blue 9 to acid yellow 23 range from 12.1:1 to 15.3:1 (Table 2). These ratios of these products 

are similar enough that the environmental exposure and toxicity of the different mixtures of the 

dyes in the four registered products will be assumed to be equivalent.

 1 USEPA. 2005. Reregistration Eligibility Decision (RED) Ecological Risk Assessment of End-Use Products 

Containing the Dyes Acid Blue 9 and Acid Yellow 23.

http://www.epa.gov/pesticides/reregistration/REDs/aquashade_red.pdf
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Aquatic dyes containing acid blue 9 and acid yellow 23 are used to color water in small bodies of 

water to inhibit growth of algae and submerged aquatic weeds on the bottom. These dyes turn the 

water an aqua-blue color, and in doing so filter out light of wavelengths that are used by plants for 

photosynthesis. Specifically, the two dyes in aquashade absorb light primarily in the red-orange 

and blue-violet parts of the spectrum, which is part of the spectrum used by aquatic plants and 

algae for photosynthesis (Lonza Corporation 2015, Spencer 1984). Light needed for 

photosynthesis is filtered out of the water, thereby reducing the amount of light reaching algae 

and submerged aquatic plants, especially in deeper water. These dyes are also used as “inert 

ingredients” in other pesticide formulations.

These two dyes are also regulated by the Food and Drug Administration (FDA) under the Food, 

Drugs, and Cosmetics Act (FDCA), and they have been approved for use as colorants for food,

drugs, and cosmetics, many of which are listed in the Household Product Database (USDHHS, 

20142

). Acid yellow 23 is a synthetic lemon yellow dye primarily used as a food coloring. A 

number of personal care, cosmetics products, various types of medications and pharmaceutical 

products may include tartrazine. Other products, such as household cleaning products, paper 

plates, pet foods, crayons, inks for writing instruments, stamp dyes, face paints and envelope 

glues, may also contain tartrazine acid blue 9, nonfood grade (acid blue 9) toilet bowl cleaner, 

laundry detergents and (i.e., the food grade Brilliant Blue FCF (Blue 1)), is often found in ice 

cream, canned processed peas, packet soups, bottled food colorings, icings, ice pops, blue 

raspberry flavored products, dairy products, sweets, and drinks. It is also used in soaps,

shampoos, mouthwash and other hygiene and cosmetics applications. In the area of soil-water 

research, Brilliant Blue is applied in tracing studies to visualize infiltration and water distribution 

in the soil (Flury and Flühler, 1994). Tartrazine is a commonly used color all over the world, 

mainly for yellow, but can also be used with Brilliant Blue FCF or Acid blue 9 to produce various 

green shades. Only the pesticidal uses (i.e., those associated with aquatic weed control) will be 

assessed here.

Chemical names, structures, and other identifiers of acid blue 9 and acid yellow 23 are given in 

Table 1.

Table 1 Chemical Nomenclature and Structure of the Aquatic Dyes Acid Yellow 23 and Acid Blue 9.

Acid Yellow 23 Acid Blue 9

Common Name as Appearing in the 

labels of the Admiral Products 

Tartrazine Erioglaucine (diammonium salt)

Empirical Formula C16H9N4Na3O9S2 C37H34N2Na2 O9 S3 (disodium 

salt)

C37H34N2(NH4)2 O9 S3

(diammonium salt)

Chemical Name(s)

4,5-Dihydro-5-oxo-1-(4-sulfophenyl)-4-

((4-sulfophenyl)azo)-1H-pyrazole-3-

carboxylic acid trisodium salt 

3-carboxy-5-hydroxy-1-p-sulfophenyl-4-psulfophenylazopyrazole trisodium salt 

N-Ethyl-N-(4-[(4-{ethyl[(3-

sulfophenyl)methyl]amino}phenyl)(2-

sulfophenyl)methylene]-2,5-

cyclohexadien-l-ylidene)-3-

sulfobenzenemethanaminium 

hydroxide inner salt, trisodium salt

N-Ethyl-N-(4-[(4-{ethyl[(3-

sulfophenyl)methyl]amino}phenyl)(2-

 2 US Department of Health and Human Service. Household Products Database.

http://householdproducts.nlm.nih.gov/index.htm
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Acid Yellow 23 Acid Blue 9

5-hydroxy-1-(p-sulfophenyl)-4-((psulfophenyl) azo) pyrazole-3-carboxylic 

acid trisodium salt

sulfophenyl)methylene]-2,5-

cyclohexadien-l-ylidene)-3-

sulfobenzenemethanaminium 

hydroxide inner salt, triammonium 

salt

Molecular Structure Only the structure of the disodium 

salt is represented. The diammonium 

salt has ammonium instead of sodium 

as the counter cation. It is the organic 

anion that is responsible for the color 

of the dye.

Synonyms 

FD & C Yellow No. 5: 

21CFR74.705 (Foods)

21CFR74.1705 (Drugs)

21CFR74.2705 (Cosmetics)

These FD&C dyes are of high purity and 

require batch certification analysis when 

used in food, drugs, or cosmetics.

CI Food Yellow 4

CI Number 15985

CAS Registry No. 1934-21-0

Sodium Salt:

Brilliant Blue

CI Food Blue 2

Erioglaucine trisodium salt

CI Food Blue 2.

CI Number 42090

CAS Registry No. 2650-18-2

Triammonium salt

Erioglaucine diammonium salt

Alphazurine

CI Blue 8

CI 42090

CAS Reg. No. 3844-54-9

FD&C Blue 1

21CFR74.101 (Foods)

21CFR74.1101 (Drugs)

21CFR74.2101 (Cosmetics)

D&C Blue 4

21CFR74.1104 (Drugs)

21CFR.74.2104 (Cosmetics)

Chemical Family Azo (monoazo) dye; Contains one azo 

group, - N=N-)

Aminotriphenylmethane dye

4. Use Characterization

Aquatic dyes containing acid blue 9 and acid yellow 23 are used primarily in small water bodies 

like ornamental ponds and small lakes, fountains and other landscaping water features, swimming 

holes, aquaculture ponds, and animal watering holes. Labels of all end-use products containing 

these dyes restrict use to water bodies that are “under complete control of the user and have little 

or no outflow.” Use is permitted in human-made water bodies, although adherence to the 

restrictions mentioned above would greatly restrict use in natural waters.
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Table 2 summarizes the use information for the four registered products containing acid blue 9 

and acid yellow 23. All products are ready-to-use liquids except for Admiral WSP, which is a 

water-dispersible pellet. Products are applied directly into the water body from the shore or from 

a boat. Labels provide instructions for users to calculate the volume of the water body in units of 

acre-feet (acres of surface multiplied by average depth in feet), and provide instructions for how 

much product to apply per acre-feet for various target concentrations of total dye. The maximum 

target concentration of total dye on all labels is 2.0 parts per million (ppm).

Table 2. Uses Site and Application Information for Aquatic Dyes Containing Acid Blue 9 and Acid Yellow 23.

Product Name

(EPA Reg. No.)

% AI Ratio Acid 

Blue 9 : Acid 

Yellow 23

Maximum 

Target 

Concentration 

of Total Dye

(ppm)

Maximum Rate 

(lb/a-ft*)

Acid Blue 9 Acid Yellow 23 Acid Blue 

9

Acid 

Yellow 23

Aquashade

(33068-1) 12.60 1.04 12.1 : 1 2.0 0.6930 0.0572

Aquashade Plus

(33068-3) 30.45 2.45 12.4 : 1 2.0 0.7316 0.0589

Admiral WSP

(67064-1) 49.72 3.27 15.2 : 1 2.0 0.6837 0.0450

Admiral Liquid

(67064-2) 15.31 1.00 15.3 : 1 2.0 0.6897 0.0451

* lb/a-ft means pounds applied per the product of surface area in acres and mean water depth in feet.

The labels do not restrict reapplication of the product. Labels of Aquashade® products state: 

“Apply in season maintenance dosages, as needed, to restore loss of color due to dilution or dye 

degradation.” Labels of Admiral® products state: “Reapply as needed during the growing 

season.”

5. Conclusions from Previous Risk Assessments

In 2005, in support of reregistration, the Environmental Fate and Effects Division (EFED) 

completed an ecological risk assessment for use of aquatic dyes containing acid blue 9 and acid 

yellow 23 to control aquatic weeds and algae (D302183, 09/26/05). In that assessment, EFED 

noted that both of these dyes are expected to be persistent in the environment, as dyes are 

generally not readily biodegradable under aerobic conditions and do not degrade via abiotic 

hydrolysis. Degradation is believed to occur by slow direct photolysis, especially in shallow water 

exposed to sunlight. The vapor pressure and the Henry’s Law Constant of dyes are very low, and 

thus little would volatilize from soil or water. Dyes such as acid blue 9 and acid yellow 23 are 

very hydrophilic (Log Kow << 1) and therefore are not likely to bioaccumulate in fish.

An aquatic risk assessment was conducted based on aquatic animals being exposed to the 

maximum target concentrations of total dye specified on the label, 2 ppm. This was compared to 

results of acute aquatic toxicity studies conducted with Aquashade®. Since exposure at all test 

levels in the acute aquatic studies did not cause any mortality, conservative RQs were calculated 

as the EEC (2 ppm) divided by the observed NOAEC from the study. RQs for bluegill sunfish, 

rainbow trout, and Daphnia magna were all less than 0.01, indicating minimal acute risk to 

aquatic animals. For aquatic plants, risk to algae and submerged macrophytes in the treated pond 

is assumed since they are the target species of the products. However, since the labels limit use to 

water bodies with little or no outlet, dye concentrations in the off-site receiving bodies were 

expected to be very low and expected to have negligible effects on non-target aquatic plants.
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A terrestrial risk assessment was conducted based on predicted exposure to mammals and birds 

drinking from water treated with Aquashade® at the maximum target concentration of 2 ppm. 

Water consumption was estimated based on allometric equations that predict daily water 

consumption for birds and for mammals based on bodyweight. For all weight classes of birds and 

mammals, acute RQs were less than 0.01, indicating minimal risk to terrestrial animals.

Calculations were also conducted to estimate exposure to birds and mammals if treated water 

were to be used for irrigation. Assuming that water containing 2 ppm were applied at a rate 

equivalent to 2 inches of irrigation, the rate of application of the dyes was determined to be 

approximately 1 lb a.i./A. A risk assessment for foliar application indicated that residues on plants 

from this rate of applications would remain well below toxic levels.

Terrestrial plants were predicted to have negligible exposure and risk from the aquatic uses of 

these dyes.

6. Environmental Fate and Transport

The environmental fate data requirements were placed in “Reserved” in 1993, depending on the 

results of the required ecological toxicity studies. Because the risk assessment did not identify 

risks to fish, aquatic invertebrates, birds or mammals, the requirement for the submission of 

environmental fate studies was waived.

3 This assessment is primarily based upon the fate 

information available from the last assessment. Open literature was reviewed for additional or 

updated information and no additional environmental fate data has been submitted since the last

assessment. The environmental fate data in this assessment are qualitative, based mostly on data 

from the open literature and on structurally-related dyes which were reviewed in the previous 

assessment (RED, USEPA, 2005). Although structure-activity relationship (SAR) estimates using 

EPI Suite/EPIWIN4 were performed, these programs do not adequately estimate physical and 

chemical properties of salts, particularly those of a large anion such as in acid blue 9 and acid 

yellow 23. A more detailed discussion concerning the chemistry and environmental fate of acid 

blue 9 and acid yellow 23 is located in the 2005 Reregistration Eligibility Decision (RED, 

USEPA, 2005). Table 3 summarizes the available information on the properties of acid yellow 23 

and acid blue 9.

Sources for data reviewed included USEPA High Production Volume Information System 

(HPVIS) Robust Summaries and Test Plan; C.I. Acid Yellow (HPVIS, USEPA, 20155

) and the

US National Library of Medicine, NIH, TOXNET (Acid blue dye 96 and Acid yellow dye7

), and 

SRC (SRC, Inc, 2015).

8

The dissipation of acid blue 9 and acid yellow 23, the two active ingredients in Aquashade® and 

Admiral®, will be due to a combination of dilution, photodegradation, biodegradation, or seepage 

from the treated pond. Hydrolysis is not expected to be an important environmental fate process 

 3 USEPA. 2005. Ecological Risk Assessment of End-Use Products Containing the Dyes Acid Blue 9 and Acid Yellow 23. (DP 

Barcode D302183; 09/26/05).

4 EPI (Estimation Programs Interface) SuiteTM (formerly known as EPIWIN). 5 http://www.epa.gov/hpv/pubs/summaries/ciacdylo/c15133tp.pdf

6 Acid Blue 9 http://toxnet.nlm.nih.gov/cgi-bin/sis/search/a?dbs+hsdb:@term+@DOCNO+5890 7 Acid Yellow 23 http://toxnet.nlm.nih.gov/cgi-bin/sis/search/a?dbs+hsdb:@term+@DOCNO+7216 8 SRC 2015. http://esc.syrres.com/fatepointer/search.asp
Page 8 of 21

since these compounds lack functional groups that hydrolyze under environmental conditions.

Volatilization from moist soil surfaces is not expected to be an important fate process because the 

compound exists as an anion and anions do not volatilize. Volatilization from water surfaces is 

also not expected to be an important fate process based upon the ionic character of these 

compounds. The vapor pressures were also extremely low.

Photochemical degradation of Brilliant Blue FCF in aqueous solution can occur in surface waters, 

such as lakes or rivers. For most dyes, such degradation is likely to progress very slowly because 

synthetic dyes are designed to be very stable to light (Porter, 1973; Pagga and Brown, 1986; 

Zollinger, 1991; as cited by Flury and Flühler, 1994).

A “dissipation half-life” of aquashade is reported to be four weeks and coloration is gradually lost 

by dilution, photodegradation and some biodegradation over time (Applied Biochemists Inc., 

1992; after Aquatic Dyes9

). However, as conditions among various bodies of water differ, there is 

a range of variation in this estimate. In one study in which Aquashade was applied to earthen 

catfish ponds with high seepage rates, it was estimated that about one-half of the dye applied 

initially was lost by seepage every 2 months (Boyd and Noor, 1982).10 They found that in a study 

that decline in Aquashade concentration in a catfish pond was primarily due to seepage 

suggesting that acid blue 9 and acid yellow 23 are quite persistent in an aquatic environment. In 

another study, the half-life of Aquashade in a test pond with no outlet was found to be about two 

months whereas in other test ponds, factors such as water outflow and evaporation affected the 

dye concentrations (White et al., 197514).

Both acid blue 9 and acid yellow 23 are expected to exist almost entirely in anion form in the 

environment; and anions generally do not adsorb more strongly to soils containing organic carbon 

and clay than their neutral counterparts and are therefore expected to be mobile in the 

environment. The mobility is supported by the use of Brilliant blue FCF as a dye tracer for solute 

transport studies (Flury and Flühler, 1994). 

Table 3. Physical and Chemical Properties of Acid Yellow 23 and Acid Blue 9 (Estimated); 

Intrinsic properties.

Information Acid Yellow 23 Acid Blue 9

Molecular Weight 534.37 (Marmion, 1991) 792.84 (disodium salt) 

782.96 (diammonium salt)

(Marmion, 1991)

Vapor pressure, at 25°C, in 

mmHg

7.43 x 10-22

there is some uncertainty in these 

estimated values, however they do not 

significantly impact the assessment as 

the vapor pressure are very low

EPI Suite, 2004; USEPA, 2004. HPVIS11

25.0 (Reported in 43503402)

2.97 x 10-42

there is some uncertainty in these 

estimated values, however they do 

not significantly impact the 

assessment as the vapor pressure 

are very low

EPI Suite, 2004

28.6 (Reported in 43503401)

 9 http://www.mass.gov/eea/docs/agr/pesticides/aquatic/aquatic-dyes.pdf

11 http://www.epa.gov/hpv/pubs/summaries/ciacdylo/c15133tp.pdf
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Information Acid Yellow 23 Acid Blue 9

Solubility in Water, 25°C 

(mg/L)

1 x 106; EPI Suite, 2004

“Completely soluble” (Reported in 

43503402)

200,000 mg/mL (or mg/L) USEPA, 

2004. HPVIS16

1.45 mg/L; EPI Suite, 2004 

“Completely soluble” (Reported in 

43503401)

Log n-Octanol/Water 

Partition Coefficient

-10.7

EPI Suite, 2004 and SRC

-10.17 SCCNFP/0786/04

-10.17 HPVIS16 c1533rr2.pdf 201-

16674B

-1.50

EPI Suite, 2004 and SRC

-0.32 SCCNFP/0787/04

Log BCF -0.54 to 0.48 (MITI, 1992; after 

Ollgaard et al. 199812.)

-

Terrestrial Fate Not biodegradable in activated sludge 

HPV c1533rr2.pdf 201-16674B

-

7. Ecotoxicity

a. Ecotoxicity Data

Available ecotoxicity data for aquatic dyes containing acid blue 9 and acid yellow 23 are 

summarized in Table 4. The data are limited to tests of acute toxicity to terrestrial and aquatic 

animals. All of the studies were classified as Acceptable. While the studies were conducted with 

one formulated product, the percentages of the two dyes in the other formulated products are

similar enough to this one that the conclusions are assumed to be applicable to all registered 

products.

Acute oral and subacute dietary studies with birds found no mortality or sublethal effects at the 

limit dose or concentration. One acute oral study with the rat also found no mortality at the limit 

dose. One out of ten rats tested fed a diet of 5,000 mg a.i./kg-diet was observed to be emaciated

and producing few feces, suggesting possible food avoidance. It is uncertain, however, if 

symptoms observed in this one animal were related to the exposure to the two active ingredients.

These results classify dyes containing acid blue 9 and acid yellow 23 as being practically nontoxic 

to birds and mammals.

In three acute aquatic tests, acute toxicity of Aquashade® is tested with bluegill sunfish, rainbow 

trout, and the waterflea (Daphnia magna). In all three tests, no mortality or sublethal effects were 

observed at the limit concentration. These results classify dyes containing acid blue 9 and acid 

yellow 23 as being practically nontoxic to fish and aquatic invertebrates.

 12 http://www.miljoindflydelse.dk/udgiv/publications/1999/87-7909-548-8/pdf/87-7909-546-1.pdf
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Table 4. Summary of Available Data on the Toxicity of Aquatic Dyes Containing Acid Blue 9 and Acid Yellow 

23 to Terrestrial and Aquatic Organisms

Species Study Type1 Test Material Source Results2

Birds

Northern Bobwhite Acute Avian Oral Aquashade®1 433367-01 LD50 > 2250 mg a.i./kg

NOAEL = 2250 mg a.i./kg

Mallard Acute Avian Oral Aquashade® 433367-02 LD50 > 2250 mg a.i./kg

NOAEL = 2250 mg a.i./kg

Northern Bobwhite Avian Dietary Aquashade® 435034-03 LC50 > 5,620 mg a.i./kg-diet

NOAEC = 5,620 mg a.i./kg-diet

Mallard Avian Dietary Aquashade® 435034-04 LC50 > 5,620 mg a.i./kg-diet

NOAEC = 5,620 mg a.i./kg-diet

Mammals

Laboratory rat Mammal Acute Oral Admiral® WSP2 452811-01

LD50 > 5,000 mg a.i./kg

NOAEL < 5,000 mg a.i./kg

(1 out of 10 rats was emaciated 

and had few feces)

Fish

Bluegill sunfish Fish Toxicity 

Bluegill Aquashade® 432975-02 LC50 > 96 mg a.i./LNOAEC = 96 mg a.i./L

Rainbow trout Fish Toxicity 

Rainbow Trout Aquashade® 432975-01 LC50 > 96 mg a.i./LNOAEC = 96 mg a.i./L

Aquatic Invertebrates

Daphnia magna Invertebrate Toxicity Aquashade® 432975-03 LC50 > 97 mg a.i./L

NOAEC = 97 mg a.i./L 1 Aquashade® is an end-use product containing 23.63% acid blue 9 and 2.39% acid yellow 23. 2 Admiral® WSP is an end-use product containing 49.62% acid blue 9 and 3.05% acid yellow 23.

While data on chronic mammalian toxicity of these aquatic dye products have not been submitted 

to the EPA, results of chronic mammalian studies with the two dye ingredients have been 

published in the open literature. Borzelleca, et al. (1990) report on lifetime chronic 

toxicity/carcinogenicity studies of acid blue 9 with rats and mice. The dye used in the studies was 

90% pure. In a two-generation rat study, dye was administered in feed to F0 generation rats for 2 

months and then in the diet to their offspring (the F1 generation) for 30 months or until death. Test 

consisted of two control groups of 120 animals each and three treated group of 120 animals 

administered dietary doses of 0.1, 1.0, or 2.0% acid blue 9. There were equal numbers of males 

and females (60/group). There were no compound-related effects on fertility, gestation, 

parturition, lactation, or pup survival. In the 2.0% group, there was a significant reduction in F1

female body weights after week 90, and a significant reduction in survival. Final mean body 

weight of females was 15% less than the pooled control mean. The NOAEC for rats was 1.0% 

(10,000 mg/kg-diet), or 631 mg/kg-BW/day.

In a one-generation mouse study, dye was administered in feed to mice for 24 months or until 

death. Test consisted of two control groups of 120 animals each and three treated group of 120 

animals administered dietary doses of 0.5, 1.5, or 5.0% acid blue 9. There were equal numbers of 

males and females (60/group). No consistent, biologically significant compound-related adverse 

effects were observed at any test level. The mouse NOAEC therefore was 5.0% (50,000 mg/kgdiet), or 7354 mg/kg-BW/day.
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Borzelleca, et al. (1988) report on two-generation chronic toxicity/carcinogenicity studies of acid 

yellow 23 with rats. One ‘low-dose” test consisted of two control groups and treated groups with 

0.1, 1.0, or 2.0% acid yellow 23. A second “high dose” test consisted of a control group and a 

5.0% treated group. In both tests, each group was composed of 60 rats per sex (120 total) in the F0

generation, and 70 rats per sex (140 total) in the F1 generation. No compound-related effects were 

noted in any treatment group in either test. The rat NOAEC was therefore 5.0% (50,000 mg/kgdiet) or 2641 mg/kg-BW/day.

b. Aquaculture Studies

Published studies have demonstrated that the dyes in Aquashade® do not exert any appreciable 

direct toxicity to aquatic plants, but merely reduce their growth through shading sunlight. A study 

by Manker and Martin (1984) found that hydrilla grown with Aquashade® and sodium azide did 

not grow significantly different than plants growing with Aquashade® alone. Since sodium azide 

is known to protect cells from oxygen radicals, this experiment indicates that Aquashade® does 

not cause oxidative damage to plant cells through creating oxygen free radicals, which has been 

proposed as an alternative mode-of-action to shading. Manker and Martin (1984) also compared 

the growth of hydrilla growing in water containing Aquashade® with plants growing in dye-free 

water, but with the light source shaded by Aquashade solutions® that were in a separate vessel 

placed between the light source and the vessels containing the hydrilla. This experiment found 

that the growth of hydrilla that were shaded by the Aquashade® solution without being in direct 

contact with the dyes was identical to the plants that were grown in Aquashade® solutions. 

Spencer (1984) found similar results with algae growth rates, which were not reduced when 

grown in shallow vessels with Aquashade® concentrations less than 5 to 10 ppm. The algal cells 

in this experiment had direct contact with the dyes, but because the water was shallow (less than 2 

cm), there was little shading of light. In contrast, in another experiment algae was grown in dyefree water that was shaded by a 1-meter column of water containing Aquashade®, again placed in 

a separate vessel locate between the test vessel and the light source. In this case, shading by 

Aquashade® concentrations of 1 and 5 ppm resulted in significantly reduced photosynthetic rates 

in the algae. 

These experiments indicate that the aquatic dyes containing Acid Blue 9 and Acid Yellow 23 do 

not control aquatic plants through any direct toxicity, but merely by filtering out light used by 

plants for photosynthesis. The lack of a toxic mode-of-action at the cellular level is consistent 

with these results of toxicity studies that show no acute toxicity in birds, mammals, fish, and 

daphnids when tested up to limit doses. Indeed, as shown by the experiment by Spencer (1984), 

the dyes will even have little effect on aquatic plankton in shallow reaches of the water column. 

Effects on aquatic plants will mainly be limited to aquatic macrophages and filamentous algae 

growing, and aquatic plants growing in deeper water. The limitation to effects only in deeper 

water would explain the finding of Ludwig (2010) that Aquashade® treatment of aquaculture 

ponds with an average depth of 50 cm had no significant effect on macrophyte weight and no 

significant effects on the growth of cyanobacteria, green algae, or euglenoid algae, nor on the 

zooplankton which feed on them. Similarly, Boyd and Noor (1982) did not find any evidence that 

Aquashade treatments effected the abundance of phytoplankton or the coverage of submerged 

aquatic plants in 1-m deep fish ponds. Therefore, even at full treatment concentrations, these dyes 

are expected to have little effect on aquatic plants in shallow water such as creeks, shallow 

streams, or wetlands. If the dyes were inadvertently released into deep water bodies, such as lakes 
Page 12 of 21

or rivers, they theoretically could have adverse effects on aquatic plants, but in these cases 

dilution would likely reduce the dye concentrations to well below effective concentrations.

Aquatic dyes containing Acid Blue 9 and Acid Yellow 23 are used in aquaculture, and thus 

research has been conducted to investigate if the dyes result in any adverse effects to the fish 

production and the aquatic ecosystems of fish ponds. Ludwig, et al. (2010) studied the effects of 

Aquashade® treatment of 0.04-ha ponds used for fish production. The ponds were fertilized with 

rice bran, and stocked with 15,000 fish fingerlings. Treated ponds received two treatments each of 

Aquashade® at a rate of 4.8 ppm. A rate higher than the label recommends was used to 

compensate for the test ponds being shallow. The pond water was monitored for water quality and 

phytoplankton and zooplankton abundance was monitored for approximately 30 days. 

Chlorophyll a concentrations were also monitored for 4 days. 

Results indicated that the Aquashade® treatments had very little effects on the fish fingerling or 

the ecosystem. There were no significant differences in the survival, abundance, or growth of fish 

between treated and control ponds. There were also no significant differences in phytoplankton or 

zooplankton with the exception of diatoms, which were at a higher concentration in the treated 

ponds during the end of the study. Chlorophyll a concentrations were significantly lower in 

treated ponds, but there was no significant difference in respiration, gross production, or net 

production. There were also little differences in water quality between treated and untreated 

ponds. There were no significant trend for unionized ammonia and pH to be higher in the control

ponds, which suggest that the moderation of algae growth may actually have some beneficial 

effects on water quality.

In an earlier study, Boyd and Noor (1982) evaluated the effects Aquashade® treatment of 1-m 

deep, 0.04-ha ponds stocked with fingerlings of channel catfish (Ictalurus punctatus). Although 

the study was flawed by an unrelated fish kill in one of the ponds, apparently resulting from a 

heavy duckweed infestation, mean fish production in the two remaining treated ponds (3,641 

kg/ha) appeared to be similar to or slightly greater than mean fish production in the three control 

ponds (3,010 kg/ha). 

Bristow, et al. (1996) compared the effects to water dyed with 2 ppm and 20 ppm Aquashade® 

versus clear water to the performance of cultured walleye larvae in fish hatchery tanks. They 

evaluated the growth and health of the fish larvae by measurements of survival, length, terminal 

weight, and gas bladder inflation. At 2 ppm, which is equal to the maximum target dye 

concentrations for all registered products, none of the performance endpoints were significantly 

different between the tanks with dyed water and those with clear water. Fish length was 

significantly reduced compared to the control in tanks with a dye concentration of 20 ppm. 

However, this concentration is 10 times greater than the maximum target concentration listed on 

registered products. Considering that accidental release of the dyes from treated waters would be 

diluted with untreated water in the receiving body, and thus be considerably less than the target 

concentration, this study indicates that fish larvae are not likely to be effected by registered uses 

of these dyes.
Page 13 of 21

c. Ecological Incidents

A query of the Ecological Incident Information System (EIIS, version 2.1) in March of 2015

identified one ecological incident of adverse field effects that was associated with use of a dye 

containing acid blue 9 and acid yellow 23. In October of 2010, a major bird and fish kill occurred 

at a country club in Lakewood County, California. An initial incident report from the California 

Department of Fish and Game (I022432-022) states that at least 150 dead birds (including 

mallards, northern shovelers, and coots) and 50 dead fish were observed at the incident site. The 

10-acre pond where the incident occurred had been treated with Admiral® Liquid (EPA Reg. No. 

67064-2), which contains 15.31% acid blue 9 and 1.00% acid yellow 23. The pond had also been 

treated with Sonar® (EPA Reg. No. 67690-4, active ingredient fluridone) and a colorant product 

called Black Onyx. The manufacturer of Black Onyx does not claim any pesticidal properties, and 

therefore the product is not regulated under FIFRA and does not have any listed active 

ingredients. While the cause of this incident is currently unknown, the very low toxicity of acid 

blue 9 and acid yellow 23 to both birds and fish make it very unlikely that they were the cause of 

the observed mortality. This incident is therefore classified as “unlikely” for these two dyes.

A review of the Avian Monitoring Information System (AIMS), which is maintained by the 

American Bird Conservancy, did not identify any ecological incident associated with the use of 

either of these dyes. 

In addition to the incidents recorded in EIIS and AIMS, the Agency maintains a database which 

stores counts of minor incidents that pesticide registrants record in quarterly aggregated incident 

reports. Ecological incidents recorded in these aggregated reports include those categorized as 

‘minor fish or wildlife’ (W-B), ‘minor plant’ (P-B), and ‘other non-target’ (ONT) incidents. A 

review of this database in May 2015 identify only a single minor fish or wildlife (W-B) and no 

plant or other non-target incident associated with use of acid blue 9 and acid yellow 23. The W-B 

incident (I025529-022) occurred in 2013 during the reporting period of March 7 to June 30, and 

was associated with use of Aquashade (EPA Reg. No. 33068-1). No other information is available 

on this incident.

8. Exposure Pathways of Concern

Based on the use pattern of aquatic dyes containing acid blue 9 and acid yellow 23, exposure to 

nontarget organisms is expected to be mostly limited to organisms living in and around the water 

bodies which are directly treated. Aquatic organisms would be exposed from ambient exposure to 

the treated water. Very limited exposure may occur to aquatic organisms in nearby natural water 

lying downhill of the treated water bodies because of the “little outflow” that the label permits for 

treated water bodies, as well as by possible overflow during heavy rain events. Spray drift, runoff, 

and atmospheric deposition are not relevant exposure routes.

Terrestrial animals living near the treated water body may be exposed by drinking from or bathing 

in the treated water body. The ecological risk assessment therefore includes an assessment of 

birds and mammals that are exposed via drinking water treated at the target concentration of dyes. 

Because the dyes are applied directly to water and readily disperses in the water column, and 

because both dyes have very low the volatility, inhalation exposure is expected to be negligible. 
Page 14 of 21

9. Status of Fate and Ecological Effects Data

Available environmental fate and ecological effects data were adequate for conducting a complete 

risk assessment for these aquatic dyes. Based on the current use pattern and target pest (i.e., 

certain aquatic plants), exposure to honey bees is not expected. Therefore, no additional data are

being requested at this time.

10. Risk Description and Characterization

a. Summary of Previous Risk Assessment Conclusions

EFED completed an ecological risk assessment for aquatic dyes containing Acid Blue 9 and Acid 

Yellow 23 in 2005 for the Reregistration Eligibility Decision (RED, USEPA, 2005). The 

registered products containing these dyes that are used for aquatic weed and algae control have 

not changed since that time with the exception that one product (Pond Care Algae Blocker, Reg. 

No. 8709-6) is no longer registered. The use rates and use restrictions have also not changed. 

Therefore, all of the conclusions of the risk assessment conducted for the RED (D302183) still 

apply. These conclusions are summarized in Section 5 of this document. In summary, the 

previous risk assessment concluded very low acute risk to terrestrial animals, fish, and aquatic 

invertebrates. Risk to terrestrial plants was predicted to be minimal because exposure from 

registered aquatic uses would be minimal. Direct risk to aquatic plants was assumed.

b. Additional Information from Published Literature

Studies on the ecological effects of the use of these aquatic dyes in aquaculture systems were 

discussed in Section 7.b. The findings of these studies support the conclusions that the registered 

use of these dyes will not cause any significant toxicity to fish or aquatic invertebrates. These 

studies also evaluated growth and developmental effects in fish, and the results indicate that the 

chronic risk to fish is also low. Finally, the studies indicate that these dyes do not exert direct 

toxicity to phytoplankton or submerged vegetation, but only inhibit their growth through the 

filtering of light. Therefore, adverse effects on aquatic plants is expected to be limited mainly to 

those living in deeper water.

c. Chronic Mammalian Risk

As discussed in Section 7, published papers provide data on mammalian chronic toxicity data of 

Acid Blue 9 and Acid Yellow 23 that are adequate for quantitative use in risk assessment. 

Because these mammalian studies were not considered in the previous risk assessment, a new 

chronic mammalian risk assessment based on these data is being included in this risk assessment. 

These studies show that both of these dyes have very low chronic toxicity to mammals. 

Borzelleca, et al. (1990) determined the chronic NOAEC of Acid Blue 9 to be 10,000 mg/kg-diet, 

or 631 mg/kg-BW/day, in the rat, and 50,000 mg/kg-diet, or 7354 mg/kg-BW/day, in the mouse.

Borzelleca, et al. (1988) found that Acid Yellow 23 to be 50,000 mg/kg-diet, or 7354 mg/kgBW/day, in the mouse. 
Page 15 of 21

A screening-level chronic risk assessment for mammals was conducted based on the NOAEC for 

the most sensitive species, and the predicted exposure from mammals consuming water treated to

the maximum target concentration of 2 ppm of total dye. The maximum concentration of the 

individual dyes is determined by multiplying the concentration of total dye (2 ppm) by the 

maximum proportion of the particular dye in any registered product. The maximum concentration 

of Acid Blue 9, which would be from use of Admiral Liquid®, would be 1.88 ppm. The 

maximum concentration of Acid Yellow 23, which would be from use of Aquashade®, would be 

0.153 ppm.

Chronic risk to mammals was assessed in the same way that acute risk was assessed in the RED 

risk assessment, except using the chronic NOAELs for the toxicity endpoint. The daily dose of 

each dye was estimated by multiplying the volume of water expected to be consumed per day by 

the maximum expected dye concentration in treated water, and then dividing by the assumed body 

weight:

Daily Dose (mg/kg-day) = Daily Water Intake (L/day) × Dye Concentration (mg/L) / Body Weight (kg)

To estimate the volume of water that mammals are expected to consume per day, the allometric 

equation for drinking water intake (equation 3-17) from the EPA Wildlife Exposure Factors 

Handbook (US EPA 1993) was used:

WI (L/Day) = 0.099 (body weight in kg) 0.90

According to the labels, the maximum water concentration of total dye is 2.0 mg/L. The 

maximum fraction of each dye relative to total dye is 0.939 for acid blue 9 and 0.0763 for acid 

yellow 23. The maximum concentration of each dye is therefore 1.88 mg/L of acid blue 9 and 

0.153 mg a.i./L of acid yellow 23. Daily doses of each dye were calculated for small, medium, 

and large mammals according to the equation given above. The daily doses were then divided by 

the chronic rat NOAEL for each dye to calculate the chronic risk mammalian quotients. Results 

of these calculations are given in Table 5.

Table 5. Chronic Risk Quotients for Drinking Water Exposure of Acid Dye 9 and Acid Yellow 23

Dye Assumed Body 

Weight (Kg)

Daily Water 

Intake

(L/day)

Daily Dose of Dye

(mg a.i./kg-day)

Rat NOAEL

(mg a.i./kg-day) Chronic RQ

Acid Blue 9

0.015 0.00226 0.280 631 0.00044

0.035 0.00484 0.257 631 0.00041

1.00 0.0990 0.184 631 0.00029

Acid Yellow 23

0.015 0.00226 0.0230 2641 8.7 x 10-6

0.035 0.00484 0.0168 2641 6.4 x 10-6

1.00 0.0990 0.0151 2641 5.7 x 10-6

All chronic risk quotients were very small (<0.001). Therefore, the chronic risk for mammals 

drinking water treated with these aquatic dyes is well below the level of concern. Combining this 

conclusion with the conclusions of the previously conducted risk assessment for exposure through 

drinking water, the EPA concludes no significant acute or chronic risk to terrestrial wildlife from 

drinking water treated with acid blue 9 and acid yellow 23.
Page 16 of 21

d. Endangered Species Assessments

In November 2013, the EPA, along with the U.S. Fish & Wildlife Service (USFWS), the National 

Marine Fisheries Service (NMFS) (collectively, the Services), and the U.S. Department of 

Agriculture (USDA) released a summary of their joint Interim Approaches for assessing risks to 

listed species from pesticides. The Interim Approaches were developed jointly by the agencies in 

response to the National Academy of Sciences’ (NAS) recommendations and reflect a common 

approach to risk assessment shared by the agencies as a way of addressing scientific differences 

between the EPA and the Services. The NAS report13 outlines recommendations on specific 

scientific and technical issues related to the development of pesticide risk assessments that EPA 

and the Services must conduct in connection with their obligations under the Endangered Species 

Act (ESA) and FIFRA. 

The joint Interim Approaches were released prior to a stakeholder workshop held on November 

15, 2013. In addition, the EPA presented the joint Interim Approaches at the December 2013 

Pesticide Program Dialogue Committee (PPDC) and State-FIFRA Issues Research and Evaluation 

Group (SFIREG) meetings, and held a stakeholder workshop in April 2014, allowing additional 

opportunities for stakeholders to comment on the Interim Approaches. As part of a phased, 

iterative process for developing the Interim Approaches, the agencies will also consider public 

comments on the Interim Approaches in connection with the development of upcoming 

Registration Review decisions. The details of the joint Interim Approaches are contained in the 

white paper “Interim Approaches for National-Level Pesticide Endangered Species Act 

Assessments Based on the Recommendations of the National Academy of Sciences April 2013 

Report,”14 dated November 1, 2013. 

Given that the agencies are continuing to develop and work toward implementation of the Interim 

Approaches to assess the potential risks of pesticides to listed species and their designated critical 

habitat, this ecological risk assessment for aquatic dyes containing acid blue 9 and acid yellow 23 

does not contain a complete ESA analysis that includes effects determinations for specific listed 

species or designated critical habitat. Although EPA has not yet completed effects determinations 

for specific species or habitats, for this preliminary assessment EPA conducted a screening-level 

assessment for all taxa of non-target wildlife and plants that assumes for the sake of the 

assessment that listed species and designated critical habitats may be present in the vicinity of the 

application of acid blue 9 and acid yellow 23. This screening level assessment will allow EPA to 

focus its future evaluations on the types of species where the potential for effects exists once the 

scientific methods being developed by the agencies have been fully vetted. This screening-level 

risk assessment for acid blue 9 and acid yellow 23 indicates potential risks of direct effects to 

listed aquatic plants from use on the registered use site. Listed species of aquatic animals and 

aquatic dependent wildlife may also be affected through indirect effects because of the potential 

for direct effects on aquatic plants upon which such species may rely. Potential direct effects on 

listed aquatic plants from the use of acid blue 9 and acid yellow 23 may be associated with 

modification of Primary Constituent Elements (PCEs) of designated critical habitats, where such 

designations have been made. Once the agencies have fully vetted the scientific methods 

necessary to complete risk assessments for endangered and threatened (listed) species and their 

designated critical habitats, these methods will be applied to subsequent analyses for aquatic dyes 

 13 http://www.nap.edu/catalog.php?record_id=18344 14 http://www.epa.gov/espp/2013/nas.html
Page 17 of 21

containing acid blue 9 and acid yellow 23 as part of completing this registration review. 

e. Endocrine Disruptor Screening Program

Aquatic dyes containing acid blue 9 and acid yellow 23 were not included in the first group of 67 

chemicals issued an order to conduct Tier 1 EDSP testing. For additional information the EDSP 

program, visit http://www.epa.gov/endo/. 

f. Clean Water Act Programs

Aquashade and Admiral (acid blue 9 and acid yellow 23) have not been identified as a cause of 

impairment for any water bodies listed as impaired under section 303(d) of the Clean Water Act15. 

In addition, no Total Maximum Daily Loads (TMDL) have been developed for Aquashade or 

Admiral16. More information on impaired water bodies and TMDLs can be found at the Agency’s 

website17. The Agency invites submission of water quality data for this pesticide. To the extent 

possible, data should conform to the quality standards in Appendix A of the OPP Standard 

Operating Procedure: Inclusion of Impaired Water Body and Other Water Quality Data in OPP’s 

Registration Review Risk Assessment and Management Process18 in order to ensure they can be 

used quantitatively or qualitatively in pesticide risk assessments. Aquatic benchmarks19 have not 

been established for these chemicals. Any data submitted or otherwise located as part of the 

registration review process may be used to update aquatic life benchmarks, if applicable. 

 15 http://iaspub.epa.gov/tmdl_waters10/attains_nation_cy.cause_detail_303d?p_cause_group_id=885 16

http://iaspub.epa.gov/tmdl_waters10/attains_nation.tmdl_pollutant_detail?p_pollutant_group_id=885&p_pollutant_group_name=P

ESTICIDES

17 http://www.epa.gov/owow/tmdl/ 18 http://www.epa.gov/oppsrrd1/registration_review/water_quality_sop.htm 19 http://www.epa.gov/oppefed1/ecorisk_ders/aquatic_life_benchmark.htm
Page 18 of 21

11. References 

Borzelleca, J.F., K. Depukat, and J.B. Halagan. 1990. Lifetime toxicity/carcinogenicity studies of 

FD&C blue no. 1 (brilliant blue FCF) in rats. Food Chemistry and Toxicology 28(4):221-

234.

Borzelleca, J.F. and J.B. Halagan. 1988. Chronic toxicity/carcinogenicity studies of FD&C yellow 

no. 5 (tartrazine) in rats. Food Chemistry and Toxicology 26(3):179-187.

Boyd, C.E. and H. Noor. 1982. Aquashade® treatment of channel catfish ponds. North American 

Journal of Fisheries Management 2(2):193-196.

Bristow, B.T., R.C. Summerfelt, and R.D. Clayton. 1996. Comparative performance of 

intensively cultured larval walleye in clear, turbid, and colored water. The Progressive 

Fish-Culturist 58(1):1-10.

Flury M. and H. Flühler. 1994. Brilliant Blue FCF as a dye tracer for solute transport studies – A 

toxicological Overview. J. of Environ. Qual. 23:1108-1112

Lonza Corporation, 2015. Colorants: Aquashade®. http://www.lonza.com/productsservices/water-treatment/non-residential-water/surface-water/colorants.aspx

Ludwig, G.M., P. Perschbacher, and R. Edziyie. 2010. The effect of the dye Aquashade® on water 

quality, phytoplankton, zooplankton, and sunshine bass, Morone chrysops × M. saxatilis, 

fingerling production in fertilized culture ponds. Journal of the World Aquaculture Society 

41(S1): 40-48.

Manker, D.C. and D.F. Martin. 1984. Investigation of two possible modes of action of the inert 

dye Aquashade® on hydrilla. J. Environ. Sci. Health, A19(6):725-733.

Marmion D.M. (1991) Handbook of U.S. Colorants: Foods, Drugs, and Cosmetics and Medical 

Devices. 3rd Ed. New York,John Wiley & Sons, Inc. 

MITI (1992). Biodegradation and bioaccumulation - data of existing chemicals based on the 

CSCL Japan. Chemicals Inspecting & Testing Institute, Ministry of International Trade & 

Industry, Japan.

Pagga, U., and D. Brown. 1986. The degradation of dyestuffs: I!. Behaviour of dyestuff in aerobic 

biodegradation tests. Chemosphere 15:479-491.

Porter, J.J. 1973. The stability of acid, basic, and direct dyes to light and water. Text. Res. J. 

43:735-744.

Spencer, D.F. 1984. Influence of aquashade on growth, photosynthesis, and phosphorus uptake of 

microalgae. J. Aquat. Plant Manage. 22: 80-84.
Page 19 of 21

USEPA, 1993. Wildlife Exposure Factor Handbook. Office of Research and Development, U.S.

Environmental Protection Agency. Washington, DC. EPA/600/R-93/187a

USEPA, 2005. Reregistration Eligibility Decision for Erioglaucine and Tartrazine (Aquashade*). 

Case No. 4010. September 23, 2005. U.S. Environmental Protection Agency. Washington, 

DC. 

White, Marla P., Hippensteel, Timothy and Lembi, Carole A. 1975. Evaluation of a water-soluble 

dye for aquatic weed control (abstract). Proceedings of the North Central Weed Control 

Annual Conference.vol. 30.

Zollinger, H. 1991. Color chemistry. Syntheses, properties and applications of organic dyes and 

pigments. 2nd ed. VCH Verlagsgesellschaft, Weinheim.
20

Appendix A

OPPIN Bibliography

Ecotoxicity

71-1 Avian Single-Dose Oral Toxicity

MRID Citation Reference

40731 Verlangieri, A.J.; Shapiro, R.; Stern, A. (1979) Evaluation of the Acute Oral 

Toxicity in Mallard Ducks of: Aquashade--Aquatic Plant Growth Control: 

Report No. T-694. (Unpublished study received Feb 27, 1980 under 

33068-1; prepared by Nutrition International, Inc., submitted by 

Aquashade, Inc., Dobbs Ferry, N.Y.; CDL:241891-A)

43336701 Campbell, S.; Beavers, J. (1994) Aquashade: An Acute Oral Toxicity Study 

with the Northern Bobwhite: Lab Project Number: 196/103A. Unpublished 

study prepared by Wildlife International Ltd. 24 p. 

43336702 Campbell, S.; Beavers, J. (1994) Aquashade: An Acute Oral Toxicity Study 

with the Mallard: Lab Project Number: 196/104. Unpublished study 

prepared by Wildlife International Ltd. 23 p. 

45192101 Pedersen, C. (2000) Acute Avian Oral Toxicity (LD50) Study with Admiral 

WSP in Bobwhite Quail: Lab Project Number: 169-001-03. Unpublished 

study prepared by Bio-Life Associates, Ltd. 38 p. {OPPTS 850.2100}. 

71-2 Avian Dietary Toxicity

MRID Citation Reference

43503403 Campbell, S.; Beavers, J. (1994) Aquashade: A Dietary LC50 Study With 

the Northern Bobwhite: Lab Project Number: 196-101. Unpublished study 

prepared by Wildlife Int'l, Ltd. 36 p. 

43503404 Campbell, S.; Beavers, J. (1994) Aquashade: A Dietary LC50 Study With 

the Mallard: Lab Project Number: 196-102. Unpublished study prepared by 

Wildlife Int'l, Ltd. 35 p. 

45200901 Pedersen, C. (2000) Acute Avian Dietary Toxicity (LC50) Study with Admiral 

WSP in Bobwhite Quail: Lab Project Number: 169-002-01. Unpublished 

study prepared by Bio-Life Associates, Ltd. 69 p. {OPPTS 850.2200} 

72-1 Acute Toxicity to Freshwater Fish

MRID Citation Reference
21

43297501 Graves, W.; Swigert, J. (1994) Aquashade: A 96-Hour Static Acute 

Toxicity Test with the Rainbow Trout (Oncorhynchus mykiss): Final 

Report: Lab Project Number: 196A/107A. Unpublished study prepared by 

Wildlife International Ltd. 39 p. 

43297502 Graves, W.; Swigert, J. (1994) Aquashade: A 96-Hour Static Acute 

Toxicity Test with the Bluegill (Lepomis macrochirus): Final Report: Lab 

Project Number: 196A/108. Unpublished study prepared by Wildlife 

International Ltd. 38 p. 

44887610 McElwee, C. (1992) Lake Colorant WSP: Acute Toxicity to Bluegill 

Sunfish, Lepomis macrochirus, Under Static Test Conditions: Lab Project 

Number: J9203007B. Unpublished study prepared by Toxikon Corp. 22 

p. 

44887611 McElwee, C. (1992) Lake Colorant WSP: Acute Toxicity to Rainbow 

Trout, Oncorhynchus mykiss, Under Static Test Conditions: Lab Project 

Number: J9203007C. Unpublished study prepared by Toxikon Corp. 21 

p.

72-2 Acute Toxicity to Freshwater Invertebrates

MRID Citation Reference

43297503 Graves, W.; Swigert, J. (1994) Aquashade: A 48-Hour Static Acute 

Toxicity Test with the Cladoceran (Daphnia magna): Final Report: Lab 

Project Number: 196A/109. Unpublished study prepared by Wildlife 

International Ltd. 38 p. 

44887612 McElwee, C.; Lintott, D. (1992) Lake Colorant WSP: Acute Toxicity to 

the Water Flea, Daphnia magna Under Static Test Conditions: Lab 

Project Number: J9203007A. Unpublished study prepared by Toxikon 

Corp. 22 p. 

Environmental Fate

No Subdivision N Environmental Fate studies were submitted, as these data requirements were 

waived in 1993. Therefore, no data from guideline studies are included in this Appendix.

Published data on the chemistry and behavior of dyes in aquatic systems was used to supplement 

the environmental fate assessment. Most of the data come from structurally related dyes. Acid 

Blue 9 and Acid Yellow 23 are also food, drugs, and cosmetic colorants regulated by the Food 

and Drugs Administration.