PC CodeNumbers: 110301(Acid Blue 9) 110302(Acid Yellow 23) 110303(Aquashade, 23.63% Acid Blue 9 and 2.39%Acid Yellow 23) DP Barcode: D302 183 

### Date: September26,2005 

### MEMORANDUM

SUBJECT:



### TO:
FROM; Ecological Risk Assessment ofEnd-Use Products Containing the Dyes Acid Blu2 9 and Acid Yellow 23 (Aquashade, 23.63% Acid Blue 9 and12.39% Acid Yellow 23; Aquashade OA, 2.36% Acid Blue 9 and 0.24% Acid Yellow 23; PondCare Algae Blocker, 2.36% Acid Blue 9 and 0.24% Acid Yellow 23; Admiral Liquid, 15.31 %

### Acid Blue 9 and 1.00%Acid Yellow 23; Admiral WSP, 49.72% Acid Blue 9 and 3.27% Acid Yellow 23) Re-registration Kendra Tyler Special Review and Reregistration Division , 

### Jim Goodyear, Biologist and Silvia C. Terrnes, Chemist I --.^I-- -*

"a-. 

# ----- Ecological Risk Branch 111 I 

### Envir~nmental Fate and Effects Divisi THRU: Daniel Rieder, Branch Chief Ecological Risk Branch I11 I 

### Environmental Fate and Effects Division

Conclusions , 

### The Environmental Fate and Effects Division has assessed the potential risk ofusing Aquashade (23.63% Acid Blue 9 and 2.39% Acid Yellow 23) and other similar end-useproducts containing Acid Yellow 23 and Acid Blue 9 dyes to control algae and submerged aquaticplants in ornamental ponds, small lakes, fountainsand fish pondslhatcheries. ~asdd on available toxicity information with a variety of species, these dyes, separately or when cordbined in various enduse products do not cause significant acute or chronic toxic adverse effecks. There is some uncertainty in this generalized conclusion given that we only havetestind ontwo ofthe representative end-use products (Aquashade and Admiral WSP). Most okthe data are from acute studieswhere a single high dose is used, and at which no mortality ofotder adverseresponses I

### were observed. One mammalian chronic study with the yellow dye also indicatedno adverse effects from long-term exposure at levelsmuch higher than expected environmental exposure. In additionto registrant submitted data, the EFED also reviewed data obtained from the open literature. Effects open literature studies were obtained through the ORD Middle Ecological Division's ECOTOX literature search and retrievalprogram managed in Duluth. The data obtained from this source supported the contention that the dyes found in Aquashade and other end-use products arepractically nontoxicto all organisms tested. Open literaturefate data were obtained through 

### an in-house literature search. The products are applied by hand, by eitherusing an eyedropper, a bottle or water soluble packets to dispense a prescribed directly into the water to achieve a concentrationsufficientto block the sunlightenergy such that plants can no longer sustainphotosynthesisresulting in death. The method of application is suchthat no movement offsitevia drift or runoff is expkcted. Therefore, the only organismsexposed are the target plants, the fish, invertebrates, aquhtic amphibians dwelling in the treated ponds or the birds, mammals, reptiles or terrestrial amphibians that drink or move into the treated water. Terrestrial plants were assumed not to be exposed under normal use conditions. One uncertainty is that none ofthe labelspreclude use of treated water for irrigation. This uncertainty is low, beuause it is assumedthese treated water bodies would not normally be used for irrigation. However, until such language appearson the label, thisroute of exposure cannot be ruled out. Noterrestrialplant data were available,however one ormore labels for enh-use products corhtaining these dyes warns that emergedplants growing in or near treated water that accidently come in contact with these end-use products they may "suffer contactburns" as stated on labels suCh as the Pond Care Algae Blocker, Aquashade OA, Aquashade, and Admiral. Therefore, EqED would assume any inadvertent exposure to terrestrial plants via irrigationor other routes would have adverse affects. The EFED estimated concentrationsin the water based on the application rates specified in the labelsto attain target concentrationsin the water body. These target concentrationswere compared with fish and invertebrate toxicity values. The target concentrations specified in the labels do not approachthe concentrationsat which no effects or mortality were observed in laboratory tests. These dyes are often used in fish bearing waters (ornamental fishponds or hatcheries) and thus it is assumed, and our analysis corroborates, that these dyes do not adversely affect fish. By inference,EFED assumes amphibians would not be adverselyaffected. These dyes are highly hydrophilic and not likely to bioaccumulate in fish. As would be expected, aquatic plants in the treated water are adversely affected (killed by intent), No 

### quantitative assessment was performed for aquaticplants. Endangered Species The.EFED also assessed the potential for Aquashade and other end-useproducts containing the ~cid'yellow 23 

### and Acid Blue 9 

### dyesto potentially cause harm to Federally' listed threatened and endangered species.

### Terrestrial Animals: Listed endangered or threatened species ofbirds, mammals and reptiles are not expectedto be affected directlybecause exposure does not exceed the endangered species LOC. However, terrestrial animalsmay be affectedindirectly if aquatic plants that are necessary for their survival are eliminated. The degreeto which this might occur depends on the extentthe treated water bodies are a critical component of their food supply. Terrestrial Plants: Terrestrial plants have the potential for exposureif water treated with Aquashade or the other dye containingend-use products are used for irrigation. If 

### irrigation is precluded fiom use on the labels,EFED concludes there is virtuallyno potential for exposure to terrestrial plants. Aquatic Animals: Listed species of fish, invertebrates or amphibians are not expected to be affected directly because the exposure does not exceed the endangered speciesLOC. 

### However, aquatic animals dwelling in treated ponds might have the potential for hamif the plants on which they depend for food and habitat are eliminated. The likelihood of this occurring depends on if any listed aquatic animal species would occur in these highly managed, oftenmanrnande, normally relatively smallwater bodies with usually no outflow, or if any, a very small amount. Aquatic Plants: Listed speciesof aquaticplants which begin their life stages as submerged might be harmed if water bodies in which they are growing are treated while they are submerged. The EFED has not fully analyzed the potential for endangered species to be harmed through use ofAquashade and other end-useproducts. Such analysis would focus on the overlap of specific specieswith potentially treated water bodies, and in the case of indirect effects, the likelihood that the treated water body comprisesenough 

### of a component of the li~ted~species food or habitat base to actually impact the listed species, if that food or habitat was lost. Significant questions raised by the assessment: 

### The followingissues were identified in the available data for dyes and products

I 

### A. Purity ofthe Dyes and Products. 

### The experimental characterizationofthe physical and chemical properties required under FIFRA was not apparently done with a 100% pure dye. The Acid ~lue 9 is reported to be 50%

### pure (43503401) and Acid Yellow 23 asbeing 28% pure (431503402). Althoughthe presence of chemical impurities affect the physical and chemical properties of a chemical substance,the extentby which they affect those ofthese dyes isubown. Some of the physical and chemicalproperties reported in these studies are not !consistent with those expected for dyes and these differencesmay be related to the unspecified impurities. For example,the reported vapor pressure is too high and likelyto be associated with volatile impurities. 2 The Aquashade product that was used in the "basic six" studies was labeled as containing 23.63 %

### Acid Blue 9 and 2.39% Acid Yellow 23. The study authors stated,

### "The test substance characterizationprovided by the sponsorindicated a purity of 13.9% azure blue dye. Theuse of the term "azure" is unclear, as it could describea color or a seriesof structurally related "azure dyes". These azure dyes are not structurallyrelated to Acid Blue 9 or Acid Yellow 23. Thereforethe exact composition and purity of the toxicant that was used in the studiesisnot certain. Theregistrant must clarify what is meant by "azure blue dye".

3. 

### The above studies were submitted in support of Aquashade (23.6% Acid Blue 9 and 2.39 Acid Yellow 23 2.39% registration It isunclear if this percent composition takes into account the purity of each dye. In addition, it is not known if the impurities in the test substances used in the characterizationofphysical and chemical properties are the same asthose declared as impuritiesin the technical andlor end-useproduct-(~onfidential Business Information).It is unclear if the purity ofthe dyes in otherproducts is the same as in Aquashade.

Label Recommendations 

### The EFED has identified confusing language in the current labels and suggeststhat these label be improved for clarity.Adding the followinglanguage is recommended.

I. Water used for irrigation It is not clear iftreated water can be used for irrigation.Ifthey are used for irrigation, plant studies (conducted with a representativeend-use product) would be required. It is recommended that all 

### of the labels clearly state that "Treatedwater is not be used for irrigation " 

### 2. Outflow oftreated water Include the following statement in all 

### of the labels: "Apply only to bodies ofwater that is under complete control of

### the user and have no outJlow"



### C. Status of Data Requirements 

### Environmental Fate Data Requirements The following SubdivisionN 

### Data Requirements may be waived for the currently registered uses and products: 1. [Abiotic] Hydrolysis (161-1) 2. [Direct] Photolysis in Water (1 61-2) 3. Anaerobic Aquatic Metabolism (1 62-3) 4 Aerobic Aquatic Metabolism (162-4) 5. 

### Mobility in SoilsISediments (1 63-1)

### 6. Bioaccumulation in Fish (165-4) Should additional uses arepetitioned andlor new products, the 

### EFED will reevaluate the need for environmental fate data. Ecological Data Requirements

No additional testing isbeing required at this time, except that if the labels are not modified to preclude irrigation with treated water, 123-1or 123-2terrestrialplant testing would be required. It is possible that the seedling emergent study requirement might be llfilled with a Tier 

### I study, however, it is likely Tier I1 testing would be required for the vegetative vigor data requirement.
Table 1. 

### Status ofecological effects data adequacy for Aquashade, dyes and other end-use products containingAcid Yellow 

### 23 and Acid Blue 9 

### dyes.
Guideline Date Requirements Are Data MRID Study Adequate forEcological Classification Risk Assessment? 71-1 Avlan Acute Oral LDso Yes Acceptable Bobwhite Quail 71

-2(a) Avian Subacute Dietary Yes Acceptable BobwhiteQuad c 71

-2(b) Avian SubacuteD~etary Mallard Duck Avian Reproduction Bobwhite Quail and Mallard Wannwater Fish Acute Toxicity LC50 Bluegill sunfish Coldwater Fish Acute ToxicityLCso Rainbow Trout Freshwater Invertebrate Acute Toxicity ECjo Water flea EstuarineMarine Acute Toxicity with SheepsheadMinnow, Oyster, Mysid Shrimp FreshwaterFish Early Life Stage Fathead minnow

Freshwater Invertebrate Life Cycle Water flea

Tier I1 Terrestrial Plant Seedling Emergence(GF 871)

Tier I1 Terrestrial Plant Vegetative Vigor (GF 871) Yes Acceptable

NA

Yes

Yes

Yes

NA

NA

Reserved

Resewed Testing not required
~u~plemental~

Acceptable
Acceptable
Exposureto marine habitats not expected

Chronic effects not expected based on mode of action and lack of effects in acute tests Chronic effects not expected based on mode of actionand lack of effects in acute tests ~eserved depending labeling precluding use of treatedwater for irrigation. If water cannotbe used for irrigation, exposureto terrestr~al plantsunlikely See aboveGuideline Date Requirements AreData MRID Study AdequateforEcological Classification Risk Assessment? Tier I1 AquaticPlant Studies 123 -2 NA Aquaticplants arethe target, therefore testing is not required Release from

treated pond to other waterbodies considered a vely low probability Mode of action suggests low likelihood 141-1 oftoxicity, method of application minimizes exposure to terrestrial insects Honey Bee Acute Contact Toxicity

### ECOLOGICAL RISK ASSESSMENT Re-registration "Acid Blue Y"(Eriog1aucine) and "Acid Yellow 23" (Tartrazine) Dyes Used together in the End-use Products Aquashade, Aquashade OA,Admiral Liquid, Admiral WSP and Pond Care AlgaBlocker for Control of Algal Growth and Other Undesirable aquatic Plants Acid Yellow 

### 23: PC CodeNumber:110302 CASRegistryNo.: 1934-21 -0 Acid Blue 

### 9 (trisodium salt) PC Code Number: 110301 CAS Regishy No.: Trisodiumsalt: 2650-18-2 Triarnmoniumsalt: 3844-45-9 FD&C Blue 1 In theTriammonium salt. the counter cation is ammonium instead of sodium. The nature of the counter cation is not relevant becauseit is the anion what is responsiblefor color of the dyes

Reviewers: Ecological Effects: James Goodyear, Biologist EnvironmentalFate: Silvia C. Termes, Chemist Secondary Reviewers: 

### Mark Corbin Stephanie 

### S yslo Brian Anderson J
Branch Chief: Daniel Rieder Ecological Risk Branch 

### I11 Environmental Fate and Effects Division

### Table of 

### Contents



### ..................................................... I. Executive Summary. 

### ..3 A. Nature of the Chemical Stressor B. Potential Risks to Non-target Organisms C. Conclusions to the Exposure Characterization D. Conclusions to the Effects Characterization E. Uncertainties and Data Gaps



### IV. 

### ................................................... Problem Formulation.. .8 A. Stressor Source and Identity 

### B. Receptors C. Assessment End-points 

### D. Conceptual Model 

### E. Measures to Evaluate Risk Hypothesis 

### F. Measures ofEcosystems and Receptor Characteristics G. Analysis Plan



### .................................................................. Analysis.. 

### -19 A. Use Characterization 

### B


### . Exposure Characterization 

### C. Ecological Effects Characterization



### ................................................... Risk Characterization.. 

### .32



### .......................................................... Literature Cited.. 

### .39
Appendices Appendix A Appendix 

### B AppenQx C Appendix D Appendix E Appendix F Appendix 

### G Appendix H Appendix 

### K Data Gaps Environmental Fate Aquatic Exposure and Modeling Ecological Toxicity and Uncertainty in Ecological Toxicity Data Status of Data Requirements Environmental Fate Bibliography Ecological ToxicityBibliography Endangered and Threatened Species Risk Quotient Calculation and InterpretationI. 

### Executive Summary A. Nature of the Chemical Stressor The chemical stressors are the dyes Acid Yellow 23 and Acid Blue 9, which are used together in several end-use products intended to control algae and other undesirable aquatic plants in artificial, confined water bodies (ornamentalponds, golf course lakes, fountains; hatcheries; swimming pools). The dyes block sunlight energy necessary for algal growth and, at the same time, impart a greenish blue coloration to the water. The two dyes are alsoregulated

I under FIFRA as "inerts" used as colorants in some pesticide formulations. In addition,they are regulated under the Food, Drug, and Cosmetic Act as colorants for food (food additives), drug formulations, and cosmetics1. The specific end-use products containingAcid Blue 9 and Acid Yellow 23 are: (a) Pond Care Algae Blocker (2.36% Acid Blue 9 and 0.24% Acid Yellow 23; EPA Reg. No. 8709-6); (b) Aquashade (23.6% Acid Blue 9 and 2.39% Acid Yellow 23, EPA Reg. NO. 3306-1); (c) Aquashade OA (2.36% Acid Blue 9 and 0.24% Acid Yellow 23; EPA Reg. No. 33068-2); (d) Admiral Liquid (15.31% Acid Blue 9 and 1.00 % Acid Yellow 23; EPA Reg. No. 67064-2); (e) Admiral WSP (49.72% Acid Blue 9 and 3.27% Acid Yellow 23; EPA Reg. No. 67064-1). Except for Admiral WSP, all the products are liquid formulations. The Agency had assigned the PC 110301to Acid Blue 9,110302to Acid Yellow 23, and 110303 to Aquashade, the mixture of 23.6% Acid Blue 9 and 2.39% Acid Yellow 23 

### (an end-use

, product). In this document, the names Acid Blue 9 and Acid Yellow 23 were used when the percentage of each dye was not specified or when individual dyes were dehcribed separately. "Aquashade" was used when referring to the test substanceused in the ecological toxicity studies. B


### . Potential Risks to Non-target Organisms Based on available toxicity data and the exposures expected when used according to the label, the Environmental Fate and Effects Division (EFED) does not believe that "Aquashade" and other products containing Acid Blue 9 and Acid Yellow 23 

# '

The specific names used for "Acid Blue 9"under the FDCA arc "FD&C Blue No.1" for the trispdium salt and "D&C" "Blue No.4" for the triammonium salt (alsoknown as erioglaucine).Both are salts of the same anion. Acid Yellow 23 is FD&C "Yellows", also known as Tartrazine.

The ecological toxicity studies were conductedwith an Aquashade end-use product containing23.6% Acid Blue 9 and 2.365 Acid Yellow 23. The PC Number assigned to the productis 110303,which has created some confusionbecause this product 1s a combinationof two dyes, each with and assigned PC Number,pose a direct acute risk to freshwater fish and invertebrates, birds, amphibians, reptiles or mammals, including endangered species. These dyes are not expected to harm terrestrial animals that drink treated water. Note that even though we do not have toxicity data on amphibians and reptiles, aquatic phase amphibians are representedby fish, and reptiles and terrestrial phase amphibians are represented by birds. It is possible that endangered terrestrial animalsmay be affected indirectly by loss of food in water bodies where treatment occurs. The extremely limited area in which this may occur, given the label-specified water bodies and the method of application (i.e. by hand) may allow a further analysisto reduce this apparent possibility to 

### be almost nonexistent. However, this analysis has not been completed. Aquashade is an aquatic herbicide and will kill algae and nontarget submerged aquatic plants. 

### If any treated habitats contained endangered aquatic plants at the time of application, effects are possible. Exposure to emerged plants and terrestrialplants is assumedto be low, resulting in little or no risk. The Environmental Fate and Effects Division is unable to assess the potential chronic risk to animals due to lack of data and therefore cannot conclude that potential chronic risk to these taxa (and the taxa for which they are surrogates) does not exist. However, because of its low acute toxicity, mode of action which is not toxicological, and because Acid Yellow 23 dye is shown not to be chronically toxic to mammals, 

### EFED does not believe that chronic effects are
I likely and therefore, chronic studies are not needed. 

### C. Conclusions- Exposure Characterization Status ofData Requirements All of the environmental fate data requirements were placed in "Reserved" in 1993, dependng on the results of the required ecological toxicity studies. Because the risk assessment did not identify risks to fish, aquatic invertebrates, or mammals, the environmental fate studies may be "Waived". The environmental fate in this assessment is qualitative,based mostly on data from the open literature on structurally related dyes. Although structure-activity relationship (SAR) estimates using EPI SU~~~IEPIWIN~ 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. For this reason, the qualitative assessment relies on open literature information. No environmental fate data from Subdivision N guideline studies are required for the present uses. If new uses and/or changes in rate or method of application or products are p~oposed, the need for new data will be reevaluated.

EPI (EstimationPrograms Interface) suiteRV' (formerlyknown as EPIWIN)Exposure 

### in the Environment Unlike the uses on food, drugs and cosmetics, the dyes are exposed to an open (but contained) aquatic environment when used as herbicides. Because the concentratedproducts are added 

### directlyto a water body, the dyes (which do not react chemically with each other) become diluted in the treated water body. Neither runoff nor spray drift are routes of exposure because a specified amount of product is directly applied to the water body to attain recommended target concentrations of the product of either 

### "1 ppm or 2 ppm", dependingon the weed to be controlled.To attain these target concentrations, the labels specify the volume of product to be added per volume of water to be treated. These maximum, target concentrations were assumedto be maintained after treatment. That is, no degradationwas assumed. The major route of dissipation of the dyes in an aquatic environmentis likely indirect photolysis, which depends of on the nature and concentrationof natural photosensitizers as well as on the geographical location where and season when the products are used. However, biotransformationunder anaerobic conditions may also contributeto the dissipation of each dye. The specific chemical nature of photoproducts and metabolitesis not known. The dyes are predominantly associated with the water column and have no potential to volatilize from water. Although the dyes are not applied to soils, they are dnlikely to volatilize from soils. Acid Blue 

### 9 and Acid Yellow 

### 23 do not have the potential to bbaccumulate in dsh. I 

### D. Conclusions- Effects Characterization Aquashadeis a formulationthat contains, as its active ingredients, yellow and a blue dye. These dyes, Acid Yellow 

### 23 and Acid Blue 

### 9 are also used as food drug and cosmetic colorants for human consumption. Their mode of herbicidal action is not ~oxicological, therefore little toxicity is expected. 

## he toxicity information available to characteriie the toxicity to wildlife includes testing with formulations such as Aquashade and Admiral WSP and also testing with the individual dyes. Data used to characterize effects include studied conductedby a registrant (testing with Aquashade and Admiral WSP) and studies reported in published literature. Published literature were obtained by the Office of Research and DevelopmentIs (ORD) Middle Ecological Division in Duluth through their literature searphed conducted as 

### part of the ECOTOX program. All testing indicates that these dyes, whether alone, or in formulations cause no toxicity to fish, aquatic invertebrates, birds or mammals at doses and concentrationsfar above those expected in the environment. No studies were required or fobnd that tested tde dyes or formulations on aquatic or terrestrial plants. In as much as this is a control agent for submerged aquatic legetation and algae, it is assumed to have an advers4 effect on aquatic plants. The label also warns that if it gets on emergent vegetation, some burning will occur. This indicates that these dyes and formulations may be toxic to terrestrial plants. However, the method of application would preclude exposure tokestrialplants, so t&city to this group of organismsis not considered necessary. If treated waters are used for irrikation, exposureto terrestrial plants may be possible and toxicity testing for terrestrial wohld be needed to assess risk.

### E. Uncertainties and Data Gaps 1. Exposure Exposure to the dyes were based on the target dilution concentrationof a directly applied volume of product to a volume of water body to attain the concentrations specifiedin the labels. (1 or 

### 2 ppm; 1or 2 mg/L). Thus, the exposure concentrationsin water are the same as the target I concentrationsin the labels. It is assumed that the dyes are stable in the water and that the target concentration remains constant. That is, routes and rates of dissipationwere:not taken into account, as data are not available. I Indirect photolysis has been identified as the major route of dissipation of the dyes in aquatic systems. However, the specific nature of transformation products fdr Acid Blue 9 and Acid Yellow 23 is not known. Although no kinetics data are availableto asless how fast each dye photodegrades, the geographical location, season, and naturelconcentrationof natural photosensitizers would control the rate of photolysis. Therefore, an uncertainty exists on the chemical nature and concentration of photoproducts.In addition, anaerobii biotransformation may also be a route of dissipation, but the chemical identity of the metabolites specific to each dye is not known.
Purity 

### of the 

### Dyes Three major issues have been identified that are associated with the purity of the dyes

, and/or test substance used in the ecologicaltoxicity studies.

a. The experimental characterization of the physical and chemical properties required under FIFRA was not apparently done with a 100%pure dye. The Acid Blue 

### 9 is reported to be 50% pure (43503401) and Acid Yellow 23 as being 28% pure (43503402).Although the presence of chemical impurities affect the physical and chemical properties of a chemical substance, the extentby which they affect those of the dyes is unknown. Some of the physical and chemical properties reported in these studies are not consistentwith those expected for dyes and these differences may be related to the unspecified impurities. The high vapor pressure reported in the studies may be that of a volatile impurity. I b. The above studies were submitted in support of Aquashade (23.6% Acid Blue 

### 9 and 2.39 Acid Yellow 2.39% registration It is unclear if this percent compositiontake into accountithe purity of each dye. In addition, it is not known if the impurities in the test substancesused in the characterization of physical and chemical properties are the same as those declared as impurities in the technical and/or end-use product (Confidential Business Information). It is unclear if the purity of the dyes in other products is the same as in Aquashade.

c. The Aquashade product that was used in the "basic six" studies was labeled as containing 23.63 % Acid Blue 9 and 2.39% Acid Yellow 23. The study authors stabed that the test substance characterization providedby the sponsor indicated a purity of 13.9% "azure blue dye". The use of the term "azure" is unclear, as it could describe a color or a series of structurallyrelated "azure dyes". These azure dyes are not sthcturallyrelated to Acid Blue 

### 9 or Acid Yellow 23. Therefore the exact composition and purity of the toxicant that was used in the studiesis not certain. The registrant must clarify what is meant by "azure blue dye". 

### 2. Effects 

### EFED has toxicity data for both formulations containing the yellow and blue dye, plus testing with the dyes separately. Tests with the Aquashade product containing 23.6% Acid Blue 9 and 2.36% Acid Yellow 

### 23 as the test substance were conducted on birds, fish and aquatic invertebrates. The Health Effects Division 

### (HED) has a rat study conducted with Admiral 

### WSP, which showed that the 

### LDS0 was equal or higher than 5,000 mg/kg. There are also mammal acute toxicity data with both the blue and yellow dyes also indicating low toxicity. No chronic studies using aquashade or other formulations with animalshave been submitted, soit is not known for certain what would happen over an extended exposureperiod. However, there is mammal chronic testing with the Acid Yellow 23 dye which indicates it does not have chronic effects on mammals. Furthermore, these dyes are used as colorants in human foods drugs and cosmetics, so long term effects from either the yellow orblue dye to mammals are not expected. While directly correlating lack of long-term mammal toxicity to birds is uncertain, given the mode of herbicidal action and low level of exposure it (is considered unlikely that long-term exposures would have adverse effects on birds. The effects

### lof chemical impurities in thedyes is not known. No studies on aquatic plants have been required, because they are the target of Aquashade and other end-use products and it is assumed that all submerged aquaticplants in the treated pond will be killed. No terrestrialplant studieshave been required, because there is no expected exposure unless treated water are used for irrigation.

the not Because of the mode of action (blocking sunlight energy that enters bodies of water) and low acute toxicity of this pesticide, it isbelieved that additional ecological toxicity studies are needed unless new uses, methods of applicationsor new products are proposed.11. 

### Problem Formulation A. Stressor Source and Distribution 1. Source and Identity of the Stressor Pond Care Algae Blocker, Aquashade, and Admiral are trade names for products containing the two dyes4Acid Blue 9 and ~cid Yellow 23 which do not react chemicallywith each other. Each product has a different ratio of the dyes (see below), but in all of the product formulations the percent of Acid Blue 9 is higher than that of Acid Yellow 23. The use pattern of the products are essentially the same in the sense that all of the products are applied 

### directly (as a volume of product or number of packets) to artificial (or natural), confined water bodies, including swimming pools (providedthat chlorine disinfectants are not used). Minimal or no outflow are specifiedin the labels because the mode of action is such that allowing treated water to outflow would eliminate the effectiveness of the products. These dyes must remain in the water column and block the sunlight energy for a sufficienttime to inhibit photosynthesis and cause the plants to die. They are not intended for use in non-confined natutal water bodies, drinking water sources, or artificial water bodies with outflow. Use of the treated water for irrigation is not precluded on the label, and this is an uncertainty because irrigation could result in exposure to nontarget terrestrial plants, and there are no tests with terrestrial plants. The specific names of the end-use products containing Acid Blue 9 and Acid Yellow 23 as the active ingredients are AlgaeBlocker and Aquashed OA (both 2.36% Acid Blue 

### 9 and 0.24% Acid Yellow 23), Aquashade (23.63% Acid Blue 9 and 2.39% Acid Yellow 23), Admiral Liquid (15.31% Acid Blue 9 and 1.00% Acid Yellow23), and Admiral WSP (49.72% AcidBlue 9 and 3.27% Acid Yellow 23). Additional information on these end-use psoducts is presented within the "Use Characterization" section. The dyes Acid Blue 9 and Acid Yellow 23 are known by several qfferent names, which may cause confusion about the chemical identity of the dye. For this reason, each one is also identified by the chemical name used by the Office of Pesticide Programs, the Chemical Abstracts Registry Number, and their synonyms (Table 1). "Acid Blue 9';could refer to either the trisodium and triammonium salts, each one having a different Che$cal Abstracts Registry Number. In most of the product labels, the blue dye is identified as "Acid Blue 9" alone, without indicating which salt is used in the formulation. Throughoutthe documen&, the names "Acid Blue 9" and "Acid Yellow 23" are used, as they are identified as such in the prbduct labels.

### In the Admiral products, they are identified as "tartrazine" for the yellow dye and as "erioglaucine" for the blue dye. The nature of the counter cation forAcid Blue 9 in the products is unclear, although the Color Index International identifies it as the triammonium salt. It is likely that the blue dye used in the products is the triarnrnonium salt. Further informatibn on the chemical
, identity of the dyes are included in Table 2. However, because it is the adion of the blue dye what exerts herbicidal activity by blocking sunlight energy, the identity df the counter cation is not relevant to the exposure and risk assessment when used as herbicidesl.

A dye is defined as a chemical compound that is capable of imparting color and that is solublein the vehicle in which it is applied.In addition to being regulated under the Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA), the two dyes are regulated by the Food and Drugs Administration (FDA)under the Food, Drugs, and Cosmetics Act (FDCA) as color additives (colorants)in food, drugs and cosmetics


### .Acid Yellow 23 (FD&C Yellow 

### 5) is a food, drug, and cosmetics colorant. The trisodium salt of Acid Blue 

### 9 is a FD&C colorantunder the name "Blue I", whereas the triarnmonium salt (Blue 4; erioglaucine)is a colorant in drugs and cosmetics 

### (D&C). Use on foods, drugs, and cosmetics require batch5 certification prior to use. While the nature of the counter cation is important forregulation under FDCA, under FIFliA the pesticide active species is the anion. It is the anion what gives color to the treated water. Therefore, for the use of these blue dyes as an algicide and aquatic herbicide for submerged plants, the nature of the counter cation is irrelevant. While the use of the dyes in food, drug formulations and cosmetics require a certification of their purity, the purity of the dyes used as an algicidel aquatic herbicideis unclear. Acid Blue 9 and Acid Yellow 23 are also regulated under FIFRA as "Inerts" used to impart color to pesticide formulations. On December 21,2004 the Agency completed the "Reassessment of the Exemptions from the Requirement of a Tolerance for the FDA-Certified FD&C Additives Blue No. 1 [Acid Blue 91; erioglaucine] and Yellow 

### 5 [Acid Yellow 23; tartrazine]" 6. This reassessment was based on structure-activity relationships (EPISuite) and does not include a discussion of transformationproducts.. However, physical and chemical ' properties and environmental fate estimates using EPIWIN are uncertain because salts of large anions are not adequately handled by EPIWIN. The Society of Dyers and Colourists and the American Association of Textile Chemists and Colorists have developed a Color Index 

## international7 standard for iddntificationof pigments and dyes. The Color Index classifies dyes and pigmentsby theircomposition. Individual pigment or dye are identified by a unique Color Index Genericpame 

### (C.I. Name) and a Color Index ConstitutionNumber (C.I. Number), although the C.I. GenericName is most commonly used.. I Expanded listings of other names used for Acid Yellow 23 and Acid Blue 9 can be found from the Unilever Center for Molecular Informatics, Cambridge University,

"Batch" is defined as "an homogeneous lot of color additiveor color additive mixtureproduced by an identified production operation, which is set apart and held as a unit for the purpose of obtaining certificationof such quantity". The "Batch Number" is the number assignedto a batch of colorant by the person who requests certification of the batch. A "Lot Number" is the identifyingnumber or symbol assigned by the FDA to a batch of color additive after certification. Note than only the trisodium salt (Blue 1)canbe used in food.

"~eassessmnt of the Exemption from the Requirement of a Tolerance for the R)A-CertifiedqlorAdditives FD&C Blue No.1, FD&C Red No. 40, and FD&C Yellow No. 5 (Tartrazine)". U.S. Environmental ProtectionAgency, Office of Pesticides Programs. December 21,2004.Acid Blue 

### 9: http://www 

### .dspace.cam.ac.uklhandle/l8 10157

### 13 Acid Yellow 23: 

### http://www.dspace.cam.ac.uklhandle/l8 1016131 Table 1 Chemical Nomenclature and Structure of the 

### Dyes

5-hydroxy-1-(p-sulfopheny1)-4-((psulfophenyl) azo) pyrazole-3-carboxylic acid trisodium salt

~olecular Structure Acid Blue 9 Erioglaucine (diammonium salt)
N-Ethyl-N-(4-[(4- {ethy1[(3- sulfophenyl)methyl]arnino)phenyl)(2- sulfophenyl)methylene]-2,5- cyclohexadien-1-ylidenel-3- sulfobenzenemethanaminium hydroxide inner salt, trisodium salt CommonName as Appearingin the labels of the Admiral Products ChemicalName(s)

i-~th~l-~-(4-[(4-{eth~l[(3- sulfophenyl)methyl]amino)phenyl)(2-sulfophenyl)methylene]-2,scyclohexadien-1-y1idene)-3- sulfobenzenemethanaminiurn hydroxide inner salt, triammonium Acid Yellow 23 Tartrazine 4,5-Dihydro-5-oxo-l-(4-sulfophenyl)-4- ((4-sulfopheny1)azo)-1H-pyrazole-3- carboxylic acid trisodium salt 3~c~~boxy-~-hy~o~y-l-p~sulfophenyl-4- p-sulfophenylazopyrazole trisodium salt

silt Only the structure of the disodium saltis represented. The diammoniurn salt has ammonium instead of sodium as the counter cation. It is the organic anion what is responsible for the color of the dye. ,sq Na+CI Food Yellow 4 CI Number 15985 CAS Registry No. 1934-21-0 Sodium Salt: Brilliant Blue CI Food Blue 2 Erioglaucinetrisodium salt CIFood Blue 2. CI Number 42090 CAS Registry No. 2650-18-2 Synonyms

Triammoniumsalt Erioglaucine diammonium salt Alphazurine CI Blue 8 CI 42090 CAS Reg. NO.3844-54-9 FD & C Yellow No. 5: 21CFR74.705 (Foods) 21CFR74.1705 (Drugs) 21CFR74.2705 (Cosmetics) TheseFD&C dyes are of high purity and requirebatch certification analysiswhen used in food, drugs, or cosmetics.

FD&C Blue 1 21CFR74.101 (Foods) 21CFR74.1101 (Drugs) 

# 1 21CPR74.2101 (Cosmetics) 

# I

Other uses of the dye-sinclude; Acid Yellow 

### 23- Dye for wool and silk; As indicatorfor chlorine estimations in biochemistry Acid Blue 9- Biological stain; Textile dye; Wood stain; Indicator

2. Physical and Chemical Properties and Environmental Fate of the Stressor 

### Physical and Chemical Properties D&C Blue 4 2lCFR74.1104(Drugs) 21CFR.74.2104 (Cosmetics) Aminotriphenylmethanedye ChemicalFamily

The most distinct characteristicof dyes is that they absorb energy8strongly within the wavelength range of the visible spectrum 

### (360 to 

### 750 nm) and that they are highly soluble in the vehicle in which they are used, in this case water. Dyes are highly conjugated systems (i.e., multiple double bonds), for which multiple resonance structures can be written. These resonance structures cause shifting or appearanceof absorption bands into the visibte spectrum of electromagnetic radiation. Multiple resonance structures increase the stability of a molecule. The color of a dye or other colored materialsis determinedby the energy (wavelength) of maximum absorption (electronic absorption) of incident light by the molecule. The observed color is determined by the wavelength region in which the substancedoe$not absorb li ht (i.e., 

# F the reflected color). In an "acid dye", the chromophoreis part of a negatike ion (anion)


### . Therefore, the color is independent of the counter cation. Table 2 summdrizes information on the light absorption characteristics of each dye.

Light is energy. The color observedin dyes (or in gems) is causedby electronic transitionsbetween I energy levels of a molecule. The color-giving absorption band is known as a chromophore ("color bearer") Azo (monoazo) dye; Containsone azo group, - N=N-)

Although Acid Blue 9 and Acid Yellow 23belong to differentchemical families,both dyes co,ntainsulfonategroups.Table 2- ElectronicAbsorption Spectra

### of Acid Yellow 

### 23 and Acid Blue 9 in Water



### I Data Source I 

## I Table 3- Physical and ChemicalProperties of Acid ~eilow 23 and Acid Blue 9



# I Marmion. Handbook of U.S. Colorants-Food, Drugs, and Medical Devices, Third Edition.,: 1991. 

# 1 Acid Blue 9 620- 639range; 

### Amax, 630

i Blue ElectronicAbsorption Spectra Wavelength of Absorption Maximum 

### (Amax, in nm) in water Reflected Color

I EPI Suite, 2004= Tolerance Reassessment Document The physical and chemical properties of Acid Yellow 23 and Acid Blue 9 are summarizedin Table 3. Acid Yellow 23 420- 430 range, 

### A,, 425

Yellow

Acid Blue 9, C37H34N2Na2 O9S3(disodium salt) C~~H~~NZ(NH~>Z 09 S3 (diammonidmsalt) 

## I 792.84 (disodium salt) (Marmion) 782.96 (diarnmoniumsalt) Reddish-violet powder (~armion)l 2.97 x IO-~~ there is some uncertainty in these estimateddalues,however they do not significantly impact the assessment as the vapor pressure are very low EPI Suite, 12004

, 28.6 (Reportedin 43503401) 1.45mgm, EPI Suite, 2004 "~om~let~l~ soluble" (Reportedin 43503401) I -1.50 

### ' EPI Suite, 2004 and SRC

## ~ I (Estimated); Intrinsic Information Empirical Formula

MolecularWeight Physical State, as pure chemicals Vaporpressure,at25" C, in mmHg

Solubility in Water, 25Tin mg/L

Log n-OctanollWater Partition Coefficient

Submitted studies= 40503401 (Acid Blue 9) and 40503402 (Acid Yellow 23); According to the stud~es, the purity of Acid Blue 9 was 50% (43503401) and that of Acid Yellow 23 was 28%.Thus, the reported physical chemical propertiesdo not reflect those of the pure materials..The high vapor pressure reported for the two dyes are hkely) to be associated with volatile impurities. I properties Acid Yellow 23 CI&~N~N~~O~SZ

534.37 (Marmion) Bright orange-yellowpowder (Marmion) 7.43 x 10‘" there is some uncertainty in these estimated values, however they do not significantlyimpact the assessment as the vapor pressure are very low EPI Suite, 2004 25.0 (Reportedin 43503402) 1x lo6;EPI Suite, 2004 "Completely soluble" (Reported in 43503402)

-10.7 EPI Suite, 2004 and SRC

SRC= 2004. SyracuseResearch Corporation. Interactive PhysicalRoperties (PHYSPROP)Database~(rno. &arch terms: FD&C Blue. No. 1,m>&C Red No. 40, FD&C Yellow No. 5. (November 28, 2004); http://www.syrres.comlesclphysdemo.htm I IDissociation Constant (pKa): Acid dyes are weak acids. The pK, values for Acid Blue 

### 9 and Acid Yellow 23 are below 4. Therefore, they are completely dissociated in the environmentally significant pH range of 

### 5 to 9. 

### Environmental Fate Parameters (Extrinsic Properties) Dyes such as Acid Yellow 

### 23 and Acid Blue 

### 9 are not readily biodegradable under aerobic conditions nor degrade via abiotic hydrolysis (no hydrolyzable groups)10,Although they absorb energy strongly in the visible range of sunlight, they appear to undergo slow direct photolysisll Direct photolysis would be only significant in clear, shallow water. But dyes have varying susceptibilityto fading when exposed to sunlight. Light fastness12 is the degree to which a dye resists fading,which varies from dye to dye. Indirect photolysis is likely to be the cause of fading in water and one of the major routes of transformationin the environment. Unlike uses in food, drugs, and cosmetics, the dyes when used as algicides, are exposed to sunlightin an open aquatic environment. Given that sunlight energy varies accordingto latitude and time of the year, their rate of fading are expected to vary depending on geographical location and season. Bioitransformation under anaerobic conditions has as also been documented for other structurally reldted dyes. Acid Blue 

### 9 and Acid Yellow 23 do not react chemically wid each other. Although environmental fate properties for Acid Blue 

### 9 and Acid Yellow 23 were estimated with Structure-Activity Relationshipsusing the EPI (Estimation Programs Interface) suiteTM (formerly known as EPIwIN)'~ ,it was considered that the estimates are not reliable because EPIWIN is not suitable for handling salts, particularlyof those ~4th large anions. Therefore, EPIWIN introduces a high degree of uncertainty for dyes. In addition, EPIWIN does not provide any information on the products that may form as the result of photoreactions or anaerobic biotransformation. The vapor pressure and the Henry's Law Constants of dyes are very low and it is unlikely that they volatilize from soil or water. Dyes such as Acid Blue 

### 9 and Acid Yellow 23 are very hydrophilic (Log Kow 

### << 1)and, therefore 

### ,they are not likely to bioaccumulate in fish.

''~~nch, D.G. "Estimatingthe Properties of Synthetic Organic Dyes", in Handbook of Property EstimationMethods for Chemicals- Environmental Health Sciences,Editedby Robert S. Boethling and Donald Mackay. Publishedby Lewis Publishers, Boca Raton, FL, 2000; Pages 447-467. And pertinent references therein.
11

Even though it has the necessary condition (i.e.,absorb energy within the spectrumof sunlight) to undergo direct photolysis that process is slow. However,they can degrade via indirect photolysis. For further information on indirect photolysisrefer to Footnote 17.
l21n artist's color language, a dye or pigment that fadesis said to be. "fugitive" and those that dy e are called "permanent colors".
l3http://www.epa.gov/opptintr/exposure/docs/episuite.htm3. Pesticide Type, Class, and Mode of Action There are five products that containboth Acid Blue 9 and Acid Yellow 23 in their formulation: (1) Care Pond AlgaeBlocker (one label); (2) Aquashade (two labels) and (3) Admiral (two labels). The products are used to control the growth of algae in ornamental ponds, I recreational man-made (closed-system ponds, such as golf course lakes), rearing lakes for nonedible fish., and fountains containing fish. The dyes do not directly kill the algae,but prevent growthby blocking sunlight energy necessary for photosynthesis and survival of the algal and other undesirable aquatic plants. Thus, these products serve as algicides or as herbicides for undesirable aquatic plants. In addition, the dyes confer a bluish green coloring to the water body. 4. Overview of Pesticide Usage These products are to be applied directlyto the water body by adding a recommended amount of the product to a specified volume of water to attain 

### 1or 

### 2 ppm according to the target weed. The extent of use, timing of application, and where it is used is not known, but geographical location and season will affect the rate of fading and frequency of application! The dose to be apply varies with the products and is specifiedin the labels and takes into account the volume and depth of the water to be treated. The specific times of application are not includedin the labels. However, some of the labels recommend that the product be applied "early in the season" (presumably Spring) and also indicate that the product can be added to an ice surfice of the water body (i.e.,before melting). 

### B. Receptors 1. Aquatic Effects For aquatic ecosystems, ecological receptors include all aquatic life (fish, amphibians, invertebrates, plants) and those terrestrial animals (e.g.,birds and mammals) that consume water andlor aquatic organisms. Since these products are directly applied to contained water bodies, any effects on aquatic life and terrestrial animals would come from consumption of water or fish in rearing lakes or fountains. Immersion in the treated water may also have effects, such as irritation or changes in color caused by the dyes. Terrestrial animalsmay' also be affected; indirectly if they depend on the aquatic plants in the treated water bodies for food. The labels for these products state that non-target aquatic plants (water lilies, hyaqinths, cattails) may suffer contact burns if the chemicals are accidentallypoured directly into them and that desirable submerged plants may alsobe affected due to reduced levels of sunlight. Aquatic organisms that depend on sunlight for survival may be affected by the reduction of sunlight. Examples include non-target plants, animals that consume nbn-t&rgetor target plants or prey on the animals that consume aquatic plants. Risk to aquatic animalsisbased on registrant submitted acute, diletary laboratory studies with aquatic vertebrates (rainbow trout and bluegill sunfish) and invertebrates (water fleas).
IThe taxa evaluated for ecological effects are presented in Table 

### 4.

Mammals Table 4. Taxa evaluated for ecological effects in screeninglevel risk assessments.

Freshwater fish Freshwater invertebrates Taxon
Bobwhite quail (Colinusvirginianus) Laboratory rat (Rattusnowegicus) Surrogate Species Used in Risk Assessment

Bluegill sunfish (Leopomismacrochirus) Rainbow trout (Oncorhynchusmykiss)) Water flea (Daphnia magna) Birds " Mallard duck (Anus playtrhynchos)

Estuarinelmarine fish Not Required because estuarine exposureis considered unlikely Estuarinelmarine invertebrates Not Required because estuarine exposure is considered unlikely Terrestrial plants Not Required because exposure to terrestrial plants is not expected providing treated water is not used for irrigation Aquatic plants and algae Not Required because aquatic plants are the target and assumed to be controlledby use of aquashade " a Birds are used as surrogates for amphibians (terrestrialphase) and reptiles. b Freshwater fish may be surrogates for amphibians (aquatic phase).

Because of the very low exposurepotential and mode of action and very restricteduse pattern, only the avian oral, avian dietary, and acute freshwater studies were required. An acute rat study was submitted to OPP's Health EffectsDivision (HED) and 

### E~D used their review as a surrogate for wild mammals. 

### 2. Terrestrial Effects Terrestrial plants are not receptors for the direct application of Aquashade to contained water. Terrestrial animals (e.g.birds and mammals) may become receptors if they consume or

. immerse in the water treated with these dyes. Risk to terrestrial animals will be based on registrant submitted acute laboratory tests with birds (bobwhite quail and mallard duck) and acute and chronic tests, with mammals to represent all terrestrial vertebrates. In addition, effects data from open literaturewill also'be considered. I 

### 3. Ecosystems at Risk All of the products are directly applied to relatively small, ornamental or decorative water bodies without outlets and are not to be used in or discharged to streams or rivers. The ecosystemsat risk are those aquatic habitats in the treated water bodies and the areas occupiedby terrestrial animals that might consume, or immerse themselves in, the treated water. The labels allow treatment of ponds in golf courses, which suggests that "natural looking" ponds may be treated. However, it is assumed that these water bodies are managed regularly for estheticreasons (i.e. to control weed population andto add color to the water body). The terminology "relatively small" is intended to differentiatethe potential use areas from extremely large water bodies (e.g. large lakes and rivers). These products areused to curtail photosynthesis in weeds and algae by blocking sunlight energy and therefore, use of these products to control aquatic plants and algae in extremely large or flowing water bodies is impractical. 

### C. Assessment Endpoints Ecological EfSects Assessment endpoints are the "explicit expressions of the environmental value that is to be protected." Defining an assessment endpoint involves two steps: 1)identifying the valued attributes of the environment that are considered to be at risk, and 

### 2) operationally definingthe assessment endpoint in terms of an ecological entity (i.e., a community of fish and aquatic invertebrates) and its attributes (i.e., survival and reproduction). Therefore, selection of the assessment endpoints is based on valued entities (i.e., ecological receptors), the ecosystems potentially at risk, the migration pathways of pesticides, and the routes by which ecological receptors are exposed to pesticide-related contamination. The selection of clearly defined assessment endpoints is important because they provide drection and boundaries in the risk assessment for addressing risk management issues of concern.

. . reproduction)of individualsand populations of birds.

2. Survival of individual mammals and reproductive effects. 3. Survival and reproduction of'freshwater fish and invertebrates 4. Survival and Growth of Aquatic Plants lb. Bobwhite quail and mallard duck subacutedietary LC50. lc. Avian reproductiveNOAEL (in this case, no avian chronic data are available,low chronic toxicityis inferred based on low chronic toxicity to mammals) 2a. Laboratoryrat acute oral L;D5~ and reproductiontest NOAEL. 3a. Rainbowtrout and bluegill sunfish acuteLCSO. 3c. Water flea acuteLC50. 3d. Fish and invertebrate chronic toxicityNOAEL. 4. Algae and vascular plant EC50 and NOAEL 5. Survival and Growth of Terrestrial Plants 5. Terrestrialmonocot and dicot vegetativevigor and seedling emergent EC25 and NOAEL LD50=Lethal dose to 50% of the test population. LCso(EC50) =Lethal (effective) concentration to 50% of the test population. EC2S=Estimated concentration at which 25% response in tested population would be expected NOAEL =Highest dose or test level at which no statistically significant effects were observed. Conceptual Model 1. Risk Hypothesis Risk hypotheses are specific assumptions about potential adverse effects (i.e., changes in assessment endpoints) and may be based on theory and logic, empirical data, mathematicalmodels, or probability models (EPA, 1998). For this assessment, the risk is stressor-linked, where the stressors are the release of Acid Yellow 23 and Acid Blue 9 by direct application into artificial and confined water bodies (i.e. no outlets) and in accordance 

## . The following hypotheses are presumed for this screening level assessment when the products are used in accordance with instructions and the precautions statedin the labels. Aquashade represents a group of end-use products containing the two dyes Acid Blue 9 and Acid Yellow 23. They are used to control weeds in contained water bodies (no outflow)by blocking sunlightto aquatic weeds and thereby inhibiting photosynthesis. They are applied directlyto these contained water bodies. The risk hypothesis is that when used as directed, Aquashade and the other end-use products may represent a risk to aquatic and terrestrial organisms." 2. Conceptual Model Water is treated directly either by pouring the product directly, or by dropping packets. I

I Aquatic plants and animals are exposed directly. Terrestrial animals may be exposedby drinkingthe treated water or immersing in it. Terrestrial plants are unlikely to be exposed unless treated water is used for spray irrigation. 

### E. Analysis Plan 1. Preliminary Identification of Data Gaps and Methods Enpironmental Fate As a first step, the current status of FIFRA's Subdivision N Data Requirements was identified for Acid Blue 9 and Acid Yellow 23. The 161-1 (Abiotic Hydrolysis), 161-2 (Direct Photolysis in Water), 162-3 (AnaerobicAquatic Metabolism), and 162-4 (AerobicAquatic Metabolism) guideline studies were "Reserved" based on low-volumeof use of these chemicals (when use as pesticides)14, but pending the results of the ecological toxicity studies (Guidelines 

### 74 through 72). Physical and chemical properties were submitted in 1994for the individual dyes 

### (MRIDNumbers 43503401 and 43503402),reviewed, and deemed acceptable However, these tv\io studies were not conducted with a high purity dye (50% for Acid Blue 9 and 28% for Acid ~kllow 23)). These two studies were submitted in support of the Aquashadeproduct of 23.6% ~(cid Blue 9 and 2.39% Acid Yellow 23. In December 2004, the Agency completed a tolerance reassessment for these two chemicals when used as inert components in pesticide formulations. ~ost of the environ mental

l4Memorandum: "Review of Phase 4 List D Package for Aquashade" (EFGWB# 93-0119,93-0120;Chemical # 110301and 110302;Case # 819437 and 819438;DP Barcode Dl84289 and D184278). Dated February 4, 1993.fate information in the document come from structure-activity relationshipsusing computational tools (EPIWIN). In addition, a "Robust Summary for Acid Yellow 

### 23 (tartrazine)15was submittedto the Agency under the High Production Volume 

### (HPV) ChallengeProgram", which also relies on EPIWIN estimates.As already indicated,EPIWIN is not a suitableestimation program for salts,particularly for salts of a large anion.Physical and chemical properties estimated by 

### EPI SuitelEPTWINhave been included in Table 

### 3 and come from the 

### 2004 tolerancereassessment document. Information from the open literaturewas includedto present a more comprehensive, but qualitative, assessmentof the environmental fateof the two dyes beyond structure-activity estimates (Refer to the Exposure Characterization- Environmental Fate and Transport section of this document). Given that the ecological risk assessment concluded that products containingAcid Blue 

### 9 and Acid Yellow 23 did not pose risk to non-target organisms, no further environmental fate data arerequired unless new uses or products are proposed. 

### Ecological toxicity Measures of effects are obtainedfrom a suite of registrant-submitted guideline studies conductedwith a limited number of surrogatd species and from open literature. The test species are not necessarily intended to be representative of the most sensitive species but rather were selectedbased on their ability to thrive under laboratoryconditions. Acute measures of effect are the concentrationsthat produce 

### 50% mortality or growth reduction in the test organisms (LCSOs and ECsos, respectively). Chronic measures are the no observed adverseeffect level or NOAEL which is the highest test level at which significant effects were not observ'ed. I 

### 2. Measures to Evaluate Risk Hypotheses and Conceptual Model

a. Measures of Exposure from Contaminated Water For these products, which are 

### directly applied to the water body in a specified volume of the liquid product or number of packets, (no spraying), the concentration of each dye depends on the amount added and the ratio of each dye in the products. The amountto be added depends on where it is used and the size and depth of the water body. Neither runoff por spray drift are routes of exposure. Therefore, simulationmodels (GENEEC;PRZM an! EXAMS) are not applicable to estimate environmental exposureconcentrations in the treated water. To estimate exposureto terrestrial animals, estimatesof water consumption were used to derive daily doses of the formulation aquashade and compared to acute toxicity endpoints. b. Measures of Effects

"Robust Summaries& Test Plans: C.I. Acid Yellow 23 (FD&C Yellow 5)", Submitted ti the Agency by the International Associationof Color Manufacturers on March 10,2004.Since Aquashade (23.6% Acid Blue 

### 9 and 2.39% Acid Yellow 23) is composed of two dyes that are never used individually as algicide/aquaticherbicide, all of the studies submitted were conducted with the end-useproduct (Refer to the Memorandum cited in Footnote 13). Studies submitted to fulfill the "basic six" requirements (71-lb avian oral, 71-2b avian dietary, 72-lb and 72-ld Freshwater fish acute, and 72-2b Aquatic invertebrate acute toxicity). Based on low toxicity and no effects observed at the highest test level in the acute toxicity test, the mode of action, the low chronic toxicity of the yellow dye demonstrated to mammals in chronic tesfing, and the fact that both of the dyes are used as food coloring for human foods and often used in other animal foods with no long-term effects detected there is no reason to believe that otlier effects (reproductive, estuarine, marine, etc.) are likely. Chronic studies are not required. For the present ecological risk assessment, 

### EFED has used the ecological toxicity data from the studies conducted with together with an acute laboratory rat toxicity conducted with AdmiralWSP submittedto 

### HED. In addition

### EFED has made the assumption that the dyes do not renresent chronic risks based on a mammalian chronic study with the Acid Yellow dye 

### 23. I

, c. Measures of Ecosystem and Receptor Characteristics Aquashade's and the labels of the other end-use products specificalll~ state that treated water bodies are not to be drained into streams, rivers or other water systems, it is assumed that exposure to these other water bodies will not occur. The aquatic organisms\thatoccur in the treated water bodies are generally assumed to have been placed there by people that manage these decorative ponds and containers. The submerged plants in the ponds are assumed to be the target organisms, sothe risk to them is not assessed. I Since it is never applied to terrestrial environments, exposure to te~strial animals assessment estimating residues on terrestrial vegetation and invertebrates was inappropriate. Instead calculations of the amount in water consumed by birds or mammals pnkingfrom the treatecl body of water were done to derive a dose of Aquashadeto birds and mammals in three size classes. I The receptors addressedby the aquatic and terrestrial risk assessments for Aquashade are summarizedin Table 4. For aquatic assessments, freshwater fish and invertebrates were studied. Estuarine/marine animals are not represented, because Aquashade is not appliedto these environments and is not expected to drain into them. 

### 111. Analysis A. Use Characterization The use characterization of the products containing Acid Blue 

### 9 and Acid Yellow 

### 23 is based on the current labels for these products. Table 

### 7 identifies these end-use'products withtheir EPA Registration Numbers, and label date. Except for Admiral 

### WSP, all of the products are liquid formulations. The products are directlypoured or droppen into water in a specified volume of the liquid formulation or number of packets. No'spraying is involved which could lead to spray drift or inadvertent exposureto terrestrial habitats. 

## able 7 Identification of the End-use Products ContainingAcid Blue 9 and Acid Yellow 

### 23 as the Active Ingredients

Product

Pond Care Algae Blocker

Aquashade OA



## I 

## I I soluble packages 

# 1

Aquashade
Admiral Liquid

Admiral WSP

The use sites and timing of application are summarized in Table 

### 8. Note that the uses are limited to confined water bodies with minimal or no outflow, but that the Admiral products has a wider range of use sites than AlgaeBlocker and Aquashade. Only the Admiral products have uses on swimmingponds, but treated swimming ponds must not be treated with a!chlorine disinfectantto avoid fading of the dyes. Likewise, no carbon filters should be used to prevent removal of the dyes from water. EPA Reg. No Registrant Label Date
8709-6 Aquarium Pharmaceuticals,Inc. February 6,2004 33068-2 Aquashade, Inc. February 23, 1993 3306-1 Aquashade, Inc. 1981 67064-2 Becker Underwood March 14,2000 67064-1 Becker Underwood August 17,2001 Composition (by weight) Type of Formulation
2.36 % Acid Blue 9 0.24 % Acid Yellow 23 Liquid formulation 2.36 % Acid Blue 9 0.24 % Acid Yellow 23 Liquid formulation

Table 8- Use Sites and Timing of Application for the End-use Products ContainingAcid Blue 

### 9 and Acid Yellow 

### 23 as the Active Ingredient I Target pests



# , ptrolof growth of many algae and underwater aquatic weeds I 'Controlof growth of many algae and dnderwater aquatic weeds 23.63% Acid Blue 9 2.39% Acid Yellow 23 Liquid formulation 15.31% Acid Blue 8 1.00%Acid Yellow 23 Liquid formulation 49.72% Acid Blue 9 3.27% Acid Yellow 23 Concentratedblend in pre-measured water

Product Pond Care Algae Blocker I vtrolof growth of many algae and underwater aquatic weeds Control algae and aquatic vegetation

I and color the dater Cbntro~ algae and aquatic vegetation and color the wkter

I

I Use Sites Ornamental Ponds and recreational man-made, closed system ponds (Golf course lakes); Rearing lakes for non-edible fish; Fountains containing fish. Time of Application I Not specified 1 "Reapply as nekded"

IIdeal Settings: Shoppingmalls Executive officeparks Botanical gardens Recreational and amusement parks Aquashade OA

Natural and manmade contained ponds and lakes including ornamental, recreational, fish rearing and fish f~ming ponds, golfcourse ponds Fountains Aquarium OrnamentalPonds Re-circulated or artificial waterscapes

Do not apply directly to streams, other natural bodies, or any body of water not under control of theuser. Do not apply to water that will be used forhuman consumption. Not specified

Admiral Liquid

Admiral WSP Recommends application before weed growing season. May be applied while ice is still covering the water body

Natural or manmade ponds, lakes, fountains, fish farms,fishhatcheries, golf courses and swimming ponds

Natural or manmade ponds, lakes, fountains,fish farms,fishhatcheries, golf courses and swimming ponds season for optimum results May be applied while ice is still Prior to or early in the weed growing season for optimum results The liquid concentrates AlgaeBlocker,Aquashade, Aquashade OA,and Admiral liquid are added directly to water in a specified volume of product according to the volume and depth of the water body to be treated. to reach a target concentration.Admiral 

### WSP is also directly applied to water in a prescribed number of packets for specific volume of thle water bodies. The resulting concentration of the products in water is fixed to "1 or 

### 2 ppm", de@ending on the target

I weed.. The dosing specified in the labels are summarizedin Table 9


### . Table 

### 9.-Dose Recommended for the End-Use Products ContainingAcid Blue 9 and Acid Yellow 23 as the Active Ingredients I 

## 1 Product 

### I Dose 

## I

Pond Care Algae Blocker Ornamentalvonds and fountains less than 2 feet deep: 8 mL ofproduct per every 100gallons of pond water (378 L); Fish rearing ponds and large ornamental ponds over 2 feet deep: 4 mL for every 100 gallons of water Maximum applicationrate, 1ppmMost of the labels containprecautionary language andlor environmentalhazard statements,but the language is not consistent across the labels. Table 

### 10 summarizes the "Precautionary Language", whereas Table 11 summarizes the "Environmental Hazard Statements" as they appear in each label.

Aquashade OA

Aquashade

Admiral Liquid

Admiral WSP

L "Small water bodies" (unspecified) One drop for each gallon of water 'Larger water bodies" 1oz per 1,000gallons of water For controlling submerged weed, the depth of water must be 2 feet Maximum application rate, 1ppm One gallon of product (1 ppm) per acre with 4 foot average depthwhen submerged weeds and algae are growing at depths >2 feet. target pests areleafy pondweed,-Chara, slender naiad, filamentous green and blue algae, spyrogyraWater milfoil. 2 ppm to prevent production of Hydrilla's tubers. The amount of the product added depends on the volume of the water body. Amount to be added (volume) per volume of water body is specifiedin the label. Regardlessof the amount added and volume of the water body, the concentrationregime in the water is : 1ppm for filamentous blue andblue algae, Chara,Leafy Pond weed, spirogyra, slender naiad, and water milfoil 2 ppm is used to prevent Hydrillatuber production after a herbicide treatment Same as Admiral Liquld

Table 10 Precautionary Language Includedin the End-use Products ContainingAcid Blue 9 and Acid Yellow 23 as the Active Ingredients

Products Pond Care Algae Blocker

Aquashade Precautionary LanguageIncluded in the Labels I I Non-target plants (water lilies,hyacinths, cattails)may suffer contact bums if material is accideniallypoured directly into them Desirable submerged plants may alsobe affected due to Aquashade OA reduced levels of sunlight I I None

Admiral Liquid Water must be under the complete control of the user and have little or no outflow (dye concentrationmust be maintained). Do not apply directly to streams, or any body of water that 

### I may be used forhuman consumption Admiral WSP Same as for Admiral Liquid ITable 11 Environmental Hazard Statements Included in the Labels of the End-useProducts Containing Acid Blue 9 and Acid Yellow 23 as the Active Ingredients

Products 

### ( Environmental Hazard StatementsIncluded in the Labels Pond Care Algae Blocker Do not contaminate domestic livestock, irrigationwater and streams with outflow.Use only in impoundedwater with no outlet andunder the total controlof the applicator. Do not use in recreationalwaters intended for swimming or in waters ApuashadeOA

I I I B. Exposure Characterization intended for edible fish Desirable plants such as water lilies may suffer contact bum if material is accidentally poured on them directly Aquashade

Admiral Liquid

Admiral WSP

1. Environmental Fate and Transport No experimentalenvironmental fate, Subdivision

### N Guideline studieswere conducted with each individual dye as the test substance or an end-use product. These studleswere placed in "Reserved" pending the results of the required ecological toxicity studies, as indicated in the "Analysis Plan" section. For consistencywith the tolerance reassessment forinerts, the environmental fate assessmentoriginally based findings on estimates from Structure-Activity Relationship (SAR) using EPISUITE (EPIWIN). However, these estimates are considered uncertain because EPIWIN is not suitable for salts,particularly those of large anions, such as in dyes, A literature search for environmental fate studies conductedwith Acid Yellow 23 and Acid Blue 9 as the test substances did not produced sufficient experimental data specificto these two dyes. Therefore,most of the environmental fate information comes from dyes structurallyrelated to Acid Yellow 23 (azo dyes) and Acid Blue 9 (triphenylmethane dyes).Fuqher information was obtained from the "Handbook of U.S. Colorants-Food, Drugs, and Medical Devices" (Marmion, 1991). Thus, most of the present environmental fate assessment uses published information about the properties of synthetic dyes and other relevant, open literature sources (Lynch, 2000; Lynch, 

### D., OPPTS. private communication,August 2005). The environmental fate assessmentis only qualitativein nature. Acid Blue 9 and Acid Yellow 23 belong to differentchemical farnilids but both contain sulfonategroups. Becausenone of these two dyes contain hydrolyzable abiotic hydrolyqis is not a degradation pathway for these two dyes. Shoreline non-target plants (cattals, water lilies) may suffer contact burn if material is accidentallypoured on them. Do not contaminatewater by cleaning of equipment or disposalof wastes. Shoreline non-target plants (cattails;water lilies) may suffer contact bum if the material is accidentally poured on them. Apply this product only as specifiedin the label. Do not make applicationswhen weather conditions favor drift fromnontarget areas. Do not apply where runoff is likely to occur. None specifiedAs chemical substances absorbing sunlight energy, photodegradation in water can occur via direct or indirect photolysis. The direct photolysis of most dyes appears to be slow and would be only significant in clear, shallow water (which couldbe the case for ornamental uses, such as in fountains exposed to sunlight). Because the presence of natural photosensitizers in the environment16, indrect photolysis has been identified as a major transformationpathway for dyes in environmental water. Besides the concentration of photosensitizers in water, the rate of photolysis would also be a function of the solar photon flux, which depends on latitude and season. For example, photolysis would be faster in the Summerthan in the Spring and faster in the Southernthan in the Northern parts of the country. Acid Yellow 23 appears to be more phbtolytically stable than Acid Blue 9, as the latter fades faster (Marmion, 1991).Photodebradation of dyes can also occur on soil surfaces. There are no experimental data that identifiesthe photoproducts of Acid Blue 9 or Acid yellow 23. However, for azo dyes (Acid Yellow 23 is an azo dye), azo bond cleavage and ph~toredox reactions occur in most cases. Potential products are aminobenzyl sulphonic acids, but their specific chemical identity and toxicity are not known. In general, most azo dyes appear to resist biodegradation under aerobic conditions. However, microbes living in an anaerobic environment can reduce azo bonds resulting in significant loss of color (Brown and Laboureur, 1983;Weber and Adarns, 1995;Jank, et al., 19R8;Baughman, 1995). Some of the products of the anaerobic transformationof azo dyes are arovhatic amines. Therefore, in the environment azo dyes can lose their color via photoredoh readtions and by anaerobic biodegradation. Anaerobicbiodegradation is most likely to occur in sedibnents. Fading can also occur by reaction with trace metals, such as zinc, tin, aluminum, iron, and bopper, mostly via redox reactions. However, fading is primarily a photochemical process.
I Dyes such as Acid Blue 

### 9 and Acid Yellow 23 are completely dissociatedin the environmentally significant pH range of 

### 5 to 9. Because the chromophoric speciesis the anion, the dyes do not adsorb strongly to soils or sediments and are

### predominantlylassociated with the wate'rcolumn. Acid Blue 9 has been long used as a hydrologicaltracer because it does not adsorb strongly on soil particulates (Mon et al., 2005; Tsai, et al., 2004) The very low vapor pressure and Henry's Law Constant of the days indicate that volatilization would not be a significantprocess. As highly hydrophilic chemicals with Log Kow I<< 1, they are not likely to bioaccumulate in fish. 2. Measures of Aquatic Exposure

a. Aquatic Exposure Modeling

l6Natural photosensitizersin environmentalwaters includedissolved organic matter (DOM),singlet oxygen 

### (loz), and hydmxy t radicals (.OH). These species are photooxidants. In additions, surfacesof semiconductingmetal oxides (such as titanium dioxide or zinc oxide) may serve as heterogeneous photooxidants and it is apotential technologyto treat wastewater containing

.dye residues. (Baran,et al., 2003). In addition, photoreactions at the surfaceof iron oxides/hydroxidesmay alsobe involved.All of these products are to be applied directlyto a confined or mostly confined water body. Therefore, the concentrationof each dye in water is only the result of direct application to water and depends on the relative ratio of the two dyes for each product and the recommended dose. Neither spray drift nor runoff are routes of exposure for Acid Yellow 23 and Acid Blue 

### 9 in, the water. A Drinking Water Assessmentwas perforrned.andit was concluded that there was nq exposure from drinking water. This assessmentwas based on the label restrictions, which indicate that treated water is not to be used as a drinking water source for humans. For the aquatic assessment, the concentrations of Acid Yellow 23 and Acid Blue 

### 9 were those target concentrations specified in the labels 

### (1ppm or 2 ppm, depending on the target weed). These concentrations are attainedby directly adding a specified volume of product per specified volume of water to be treated (i.e., by dilution of the product into a larger volume of water). It should be noted that the dosing languagein some labels (e.g., Aquashade OA ) are vague ("one drop") in indicating the volume of product to be added to the water body. It is unclear if the specified concentrationis in terms of percent of product (i.e., the two dyes and inerts) nor the purity of the dyes are taken into account.. The concentrations resulting from direct application are summarized in Table 

### 12. Because there are no kinetics data for indirect photolysis or anaerobic biodegradation, it is assumedthat these concentrations remain constant. For Acid Blue 

### 9, the estimated concentrations donot take into account the type of counter cation (i.e.,no molar fraction correction, given the uncertainty in the identity of the salt). Even though the environmental concentrations were estimated for each individual dye, there are no ecologkal toxicity for the individual dyes because the test substance in all of the ecological toxicity studies was an end-use Aquashade product containing 23.6% Acid Blue 

### 9 and 2.39% Acid Yello~ 23 of uncertain puiity. Therefore, an assessmentbased on each individual dye was not possible. The ecological toxicity of products with a higher percentage of each individual dye andorhifferent ratio of the dyes than Aquashade may potentiallybe under represented. These products are Admiral Liquid and Admiral 

### WSP. Table 12 Maximum Concentrationsof Acid Blue 9 and Acid Yellow 23 Expected from Direct Application ofEach Product at the Label at Rates Set in the Labels and assuming +00% purity of each dye (Source: Product labels) ,

Product
Aquashade OA Aquashade Pond Care AlgaeBlocker Admiral Liquid Application Rate of Product as target concentration, ppm; 1 2 1 2 1 1 2 Concentration of Acid Blue 9,in mg1L 0.024 0.048 0.24 0.48 0.024 0.15 0.3 - Concentration of Acid Yellow 23, in mg/L

I 0.0024 0.0048 0.024 0.048 0.0023 0.01 0.02 IAdmiral WSP 1 0.497 (0.5) 0.033

I b. Aquatic Exposure Monitoring and Field Data The use of monitoring data and/or field data is not applicableto these dyes when used to control algal growth or other undesirable aquatic plants in confined, with no or minimal outlet
, water bodies. However, these dyes also have industrial applications and have been detected in waste water, sewage systems, and water bodies17.Ecological risk from those sources of potential exposure are not addressed in this document.

c. Measures of Terrestrial Exposure 3. Terrestrial Exposure Modeling Routinely, exposure to birds and mammals feeding on a treated field is estimatedby i modeling residues on terrestrial vegetation and invertebrates using the Terrestrial Residue Exposure (TREX) simulation model. Water treated with Aquashade or any of the other products is not expected to reach terrestrial vegetation or invertebrates. Because exposure to the dyes from residues on terrestrial food items is not expected, the use of terrestrial exposure models is not applicable. An exception could be if treated water was used for irrigation. If this happened, it is possible the dyes in these formulations would get on terrestrial food items. However, given the low toxicity, it is unlikely to result in direct acute risk to terrestrial animals.

a. Terrestrial Exposure via Consumption of Contaminated Water Modeling To determine the exposure of mammals and birds to "Aquashade" via consumption of contaminated water, a single daily dose of Aquashade was estimated using the calculatedvolume of water that birds and mammals are expected to consume per day and the concentrationof Aquashade in water as follows: Daily Exposure (mg) = Daily Water Consumption (L) x Water Concentration(mg/L). To estimate the volume of water that mammals and birds are expected to consumeper day, allometric equations from the EPA Wildlife Exposure Factors Handbook (US EPA 1993) I were used. For birds, the daily water consumption 

### (L) was calculated using the equation (US EPA 1993, Equation 

### 3-15,p. 3-8, for all birds): 

### L 

### = a 

### (body weight in 

### kg) B, 

### where 

### a 

### =0.059 

### and 

## P =0.67

l7 See, for example, "Background Document for Identification and Listing of Deferred Dye and Pigment Environmental ProtectionAgency, Office of SolidWaste, Washington, D.C.,June 1999.

26For mammals, the daily water consumption 

### (L) was calculated as using the equation 

### (US EPA 1993, Equation 3-7, p. 3-10, for all mammals): 

### L 

### = a 

### (body weight in kg) 

### P, where 

### a =0.099 and f3 

### =0.9 The maximum concentration (weightto volume) of Aquashade in water after application at the labeled rate is 2 mg/L. As summarizedin Table 13, a single daily exposure via drinking water containing 2 mg/L Aquashade was calculated for three weight classes for birds (20, 100, and 1000g)and mammals (15,35, and 1000g).

1,000 0.059 0.119 Mammals 15 0.0023 0.004 35 0.0048 0.01 1,000 0.099 0.1

a EstimatedDaily Exposure (mg) = Daily water consumption (L) x Water concentration 2 ma).


### b. Residue Studies There is no terrestrial exposure from this use pattern. Therefore, residue data are not necessary. C. Ecological Effects Characterization.
I This documentis a screening level evaluation of the potential ecological risk from the end-use products that contain Acid Yellow 23 and Acid Blue 9 dyes, and are representedby,a formulation referred to as Aquashade. All of the products are composed of two dyes that are never used individually as an algicidelaquaticherbicide. These products are applied drectly;to confined water bodies and never to terrestrial or aquatic ecosystems. Available data are from tests conducted with either a formulation containing Acid Yellow 23 and Acid Blue 9, or with the dyes separately. All of the toxicity studies on birds, fish and aquatic invertebrates submittedby the registrant were conducted with the end-use product Aquashade (23.6% Acid Blue 

### 9 and 2.39% Acid Yellow 23) and not with the individual dyes. The studies were submittedto support the registration of this "Aquashade" end-use product (EPA Reg. No. 33068-1). Through searches conductedby the ORD Middle Ecological Division at Duluth under their ECOTOX literature

27

### searchprogram, ecological toxicity studies conducted with the individual dyes were located. The results ofthese published literature data supportedthe results of, and did not indicate greater toxicity than, or effects other than the registrant submitted tests, therefore, they were not used to estimaterisk in this assessment. A laboratory rat study was conducted with the Admiral WSP 

### product as the test substance (49.62% Acid Blue 9 

### and 3.05% Acid Yellow 23). There is an uncertainty in the purity ofthe test substance used in the ecological toxicity studies.However, considering the low exposurelevel relative to even the lowest assumedpurity, 

## - this uncertainty does not cause doubt inthe conclusions. The studies submitted to hlfill the "basic six" requirements (an avian oral, two avian dietary, two fish acute, and one aquatic invertebrateacute toxicity) found very low toxicity (practically nontoxic and slightly toxic). A study on the laboratory rat found an LD50 equal to or higher than 5,000ppm. Table 14 summarizesthe acute data for the tested speciesused to assess risk.Aquashade Bobwhite quail Acute Avian Oral 433367-01 LD50 & acuteNOAEC 2 2250 mgm Mallard duck Acute Avian Oral 433367-02 LD50 & acute NOAEC 2 2250 mgm Bobwhite quail Avian Dietary 435034-03 LC50 & acute NOAEC 25,620ppm Mallard duck Avian Dietary 435034-04 LC50 & acuteNOAEC 2 5,620ppm
Bluegill sunfish Fish Toxicity Bluegill 432975-02 LC50 & acute NOAEC 2 96mgL

Rainbow trout Fish Toxicity Rainbow Trout 432975-01 LC50 & acute NOAEC 2 96 mg/L
Daphnia magna Invertebrate Toxicity 432975-03 LC50 & acute NOAEC 2 97 mg/L

Test Material Admiral WSP Laboratory rat Wild Mammal Acute 45281 1-01 LD50 ci & 9 r 5,000 mglkg

All were perfo~~ned with the Typical End-use Product, Aquashade or AdmiralWSP 

### * The results refer to the concentration of the end-use product. Birds and mammals are expected to drink from ponds that have been treated with the end-use products. 

### EEED calculated an estimated dose that they would receive from that water

I (see Table 10). 

### 1. Terrestrial Effects Avian effects The avian oral acute toxicity studies for both the bobwhite quail and' the mallard duck found LD50's and NOAECs equal to or greater than 2,250 mg/kg. The avian dietary toxicity studies found LCsosand NOAELs equal to or greater than 5,620

### ppm. Aquashade is considered to be practically nontoxic to birds. Higher tiered studies are not required.

Mammalian effects An acute toxicity study that used Admiral WSP with the laboratoryrAt

# WAS reviewed by OPP's Health Effects Division found an LDsoequal to or greaterthan 5,000 hg/kg. EFEDconsidersthis to be practically nontoxic. The Health EffectsDivision wrote that, 

### ".

. 

### .this formulationcontained 49.62 % Acid Blue dye and 3.05% Acid Yellow dye (presumably Acid Blue 9 [Erioglaucine],and Acid Yellow 23 [Tartrazine],respectively). A chronicmammal toxicity test with Acid Yellow 23 indicatedlow likelihood of adverse chronic effects at environmentally relevant exposurelevels. 

### Calculating Daily Dose Terrestrialanimals drink water fromponds and otherbodies and may be affectedby the Aquashade or any of the other end-useproducts in the water. Exposure estimates for terrestrial animals via drinkingcontaminatedwater were expressed in terms of a single daily dose (mg) for three different body weight classes. All of the exposureswere less than 0.2 mg a.i./day. Birds The weight classes for birds were 20, 100,and 1,000g. To obtain the toxicity value for a daily dose (mg of Aquashade) for each weight class for birds termed the "derived median dose," the acute oral 

### LDso(22,250 mgikg-bw) was multiplied by body weight (kg). The calculated daily doses for birds for each weight class are summarized in Table 13.



### Mammals Exposure estimates for terrestrial animals via drinking of contaminatedwater were expressed in terms of a single daily dose (mg) for three differentbody weight classes (15,25, and 1,000 g). Therefore,to obtain the lethal dose (mg of Aquashade) for each weight class for mammals, the acute rat oral LD50 value of 25,000 mglkg-bw 

### (MRID 45281 1-01)was multiplied by body weight (kg). The calculated daily doses for mammals for each weight class are summarized in Table 15.



### Table 15. 

### Acute toxicity values for birds and mammals drinkingfrom contaminated water, expressed in terms of a singleAquashade dose (mg). Animals Body Weight (kg) LDSO Derived Median (mglkg-bw) Lethal Dose (mg) a

Birds

1.000 12,250 22,250 0.015 25,000 275 Mammals 0.035 25,000 2 175 1.000 25,000 25,000

a DerivedMedian LethalDose (mg)=body weight (kg) x LD~o (mglkgbody weight).Terrestrial Plant efSects The method of application,i.e. pouring and dropping specific volumes or packets into the target water body, is not expected to result in drift or runoff resulting in exposure to terrestrial plants. Since it will not be applied to terrestrial plants no plant studies were required. One uncertainty exists, however, in that not all the labels preclude use of treated water for irrigation. I If it was used for irrigation, the potential routes of exposure would be drift and/or runoff. To assess risk to terrestrial plants, tier 

### 1or tier 

### 2 terrestrialplant testing would be used. If the labels

I stated that treated water must not be used for irrigation, such testing would be unnecessary. Some of the labeling indicates that emergent plants may experience burning if exposed to the dye foimulations. Based on this, it is assumedthat any inadvertent exposure to terrestrial plants might cause adverse effects. 

### 2. Aquatic Effects The registrant submitted studies that determined the toxicity of Aquashade to aquatic animals. The bluegill sunfish and rainbow trout studies found LCSOs and NOAECs equal to or greater than 

### 96 mg/L. This level is considered by EFED to be slightly toxic. The level for being considered practically nontoxic is 

### 100

### mg/L for aquatic animals. The aquatic invertebrate (Daphnia rnagna) study found that the LCso and the NOAEC were equal to or greaterthan 

### 97 mg/L. This level is considered to be slightly toxic. No further studies are required. Aquashade, Admiral, and AlgaBlocker are algicides or herbicides for undesirable aquatic plants. They are not applied to terrestrial environments nor if treated water used for irrigation. Because submerged aquatic plants are the target species and it is assumed that all submerged plants will be killed, no aquatic plant studies were required. 

### 3. ECOTOX and other Published information The Agency also obtained summary toxicity information from published literature found as part of the ECOTOX program maintainedby the EPA Office of Research and Development Middle Ecological Division (MED) at Duluth. This program regularly searches open literature for toxicity information. OPP requested of MED Duluth to search its holdings for toxicity information on aquashade and the dye components of the formulations being assessed for reregistration. Toxicity information was located on mammals and aquatic animals from tests with either of the dyes. In all cases, the test results located through ECOTOX corroborated the findings of the registrant submitted data, and did not indicate effects unforeseen from the registrant studies. Neither did these published literature studies indicate adderse/effects to organisms other than those tested by the registrant. None of the data located thrbugh ECOTOX were used in the assessment. See Appendix 

### F for a summary table of data locat{d from ECOTOX.

### IV. Risk Characterization

, Acid Yellow 23 and Acid Blue 

### 9 are applied together via the end-use products Aquashade Admiral Liquid. Admiral WSP, and AlgaBlocker. These products are applied to confinedwater bodies or water bodies with minimal outflow. None of the productshave terrestrial uses and would not directly contactterrestrial food items for birds and mammals. Therefore, the RQs based on dietary exposure from Aquashade residues on food items were not calculated. However, it could affect them 

### via drinking water. Thus, all of the potential risks of Acid Yellow 23 and Acid Blue 

### 9 are associated with direct exposure of terrestrial animals through drinking water or immersion in the treated water. In addition, aquatic organisms dwelling in treated water are also exposedto the dyes (referto "Conceptual Model") Risks were estimated based on "environmental" concentrations in a water body resulting from direct application of the product(s) that is, a dilution concentrati.onbased on the application rates indicated in the labels (see Table 10). It must be kept in mind that the characterizationof effects was based in terms of the combined two dyes for the Aquashade enduse product (23.6% Acid Blue 9 and 2.36% Acid Yellow 23) used in the ecologicaltoxicity studies and not for the individual dyes. A. Risk Estimation - Integration of Exposure and Effects Data 1. Birds and Mammals Exposure to birds and mammals was expectedprimarily through consumption of drinking water treated with the dyes. Birds are surrogates for reptiles and terrestrial phase amphibians because it is assumed for screening level assessment purposes that reptiles and terrestrial phase amphibians are not more sensitiveto oral toxicity than birds. 

### Acute Risk Quotients (RQs)via Consumption of Contaminated Water Acute RQs for birds and mammals exposed to Aquashade via consumption of contaminatedwater were calculated for each of three body weight classes using the daily exposurevalue expressed as milligrams of Aquashade (Table 13)and the toxicity value expressed in terms of milligrams of Aquashade (Table 14). The RQs are summarized in Table 16.1,000 0.12 2 2,250 < 0.01 Mammals 15 < 0.01 2 75 < 0.01

" Estimated Environmental Exposure (EEC) = Daily water consumption (L) x Water concentration(mgL). I Rounded to two decimal places. Derived Median LethalDose (mg) =body weight (kg) x LDso(mg/kg body weight). AcuteRQ =Estimated Environmental Exposure 1 Concentration Derived Median LethalDose RQs are below the Levels of Concern (LOCs) for acute risk (LOC 0.5), acute restricted use (LOC 0.2), and acute endangered species (LOC 0.1). Acute RQs are below the Levels of Concern (LOCs) for endangered species ( 0.1). 

### Use of Treated Water for Irrigation

I While it is considered highly unlikely that treated water would be used for irrigation, given the kinds of water bodies treated, if this happened, exposure to birdsand mammals might occur through ingesting terrestrial food items that had been exposed to the water. Given the low toxicity of the dyes, and the relative low concentration in the treated water (no more than 

### 2 ppm), it is unlikely that this route of exposure would cause &rect toxicity to birds or mammals. If 

### 2 inches of irrigation water containing 2 ppm were sprayed onto terrestrial vegetation, the resulting "application rate" of the dyes would be approximately 1Ib ailacre. Applying 1lb ailacreto short grass would result in residues no greaFer than 240 ppm. For a small (15g) mammal that eats close to its body weight this would still resblt in a dose per animal much less than the lowest toxicity value for mammals. See appendix

### E forlcalculations. 2. Non-target Aquatic Animals and Plants Aquashade and any of the other end-use product containing Acid Blbe 

### 9 and Acid Yellow 23 are only applied directly to ponds and other aquatic systems without an outlet. It is not expected to come in contact with non-target aquatic animals outside of those systems. This is a screening level assessment of the acute risk of using ~~uashade, therefore, the highest EECs (2 

### mgL; for Admiral Liquid and Admiral WSP) and the lowe'?ttoxicity values were used. The concentration in the treated systems will be higher than any body of water that

I 33 I

I

### accidentallyreceives (by flooding,etc.)

### a dose ofAquashade. A summary of aquatic acute RQs is presented in Table 17. ble 

### 17. Estimated acute risk quotients 

### (RQ) for 

### aquatic animals exposedto Aquashade. this screening analysis only the highest Estimated Environmental Concentration

### (EEC

I (mgJL) Bluegill sunfish 2 96 mg/L <0.01 none

, Rainbowtrout 2 96 mg/L KO.01 none Daphnia magna 2 97 mg/L 10.01 none


### Fish are used as surrogated for aquaticphase amphibians, so lack ofrisk to fishresults in 

### a conclusion oflow risk to amphibians. 1 

### Aquatic Plants: This is a screening level assessment based on an Aquashade product. Aquashade, AlgaeBlocker, and Admiral products are aquatic algicides or herbicides for undesirable aquatic plants and are expected to kill all submerged aquaticplants. However, since it is only applied to ponds with little or no outlet, it isnot expected to come into contact with nontarget aquaticout side of the target pond. All of the submergedplants inthe pond are considered to be targets. Therefore, the RQs were not calculated for aquatic plants. Ifthe water treated by these products were used for irrigation, it is unlikely the concentration of dyes in the runoff from such irrigationwould be sufficient to cause effects to aquatic plants in receiving water bodies. 3. 

### Non-target Terrestrial Plants in Dry-land and Semi-aquatic Environments Terrestrial plants growing in dry-land and semi-aquaticenvironments are not expectedto be exposed to the end-use products, since they products are poured, or appliedwith an eyedropper into confined water bodies or water bodies with minimal outflow. Because the labels do not allow application to terrestrial ecosystems, a terrestrialplant risk characterization was not performed. An uncertainty exists in that the labels do not preclude use oftreated water for irrigation. Ifthis happened, there is a potential for adverse effects to terrestrial plantsbecause some labels indicate that if emergedplants are exposed to the formulations, burning would be expected. B. 

### Risk Description 

### - Interpretation ofDirect Effects Aquashade does not exceed the level of concern for animals with which it is expectedto come in contact. Because of the method of application,no drift or runoff onto terrestrial habitats is expected precluding exposure to terrestrial plants, and to food items consumed by terrestrial animals. Thus, the risk hypothesis is proven wrong for these organisms. The mode ofherbicidal action and intended use will result in adverse effectsto target aquaticplants, therefore the risk hypothesis that these formulationsadversely affkct aquaticplants is correct. However, in all cases, the treated water body is treated for the express, reason of

### controlling (eliminating)submerged aquatic plants therefore, this effectis not interpreted as an adverse ecological effectsunless, as discussed below, there are endangered aquatic plants in these treated water bodies.

C. 

### Endangered and Threatened Species The use of these dyes may affect endangered aquatic plantspossibly occurring inponds where this might be used. It may alsohave the potential to affect endangered animal species which depend on aquaticplants in these managed water bodies for food and/or habitat. The potential for this direct effects to listed aquatic plants, and indirect effectsto listed animals is very limited because of the method of application (i.e.by hand), and the kinds ofwater bodies treated (golf course ponds and othermanaged ponds) where keeping the water free of all vegetation is aestheticallydesirable. These water bodies do not typically have outflow to streams or rivers, which would preclude endangered fish and invertebrates from entering. However, endangered amphibians may enter such water bodies, and the loss of plants that serve as food and shelter for thbse amphibianshave the potential indirectly affectthese organisms. 1. 

### Action Area For listed species assessmentpurposes, the "action area" is considered to be the area affected directly or indirectlyby the Federal action and not merely the immediate area involved in the action. At the initial Level I 

### screening assessment, broadly described taxa are considered and thus the screening conservatively assumes that listed species within those broad groups are colocated with the pesticide treatment area. Thismeans that terrestrial plants and wildlife are assumed to be located on or adjacent to the treated site, and aquatic organismsare assumed to be located in a surfacewater body adjacent to the treated site. The assessment also assumes that the listed species are located within an assumed area that has the relatively highest potential exposure to the pesticide, and that exposures are likely to decreasewith distance from the treatment area. Aquashade, Admiral, and AlgaBlocker are used in ornamental ponds, recreational manmade ponds, closed system ponds, golf course lakes,rearing lakes for fish, fountainscontaining fish, and recirculated or artificialwaterscapes. Therefore, these products are only applied to confined water bodies. 

### 2. Taxa Potentially at Risk In this assessment, it is concluded that the only speciespotentially at risk are aquatic plants from direct effectsifthey occur in treatedponds and lakes, and aquatic or terrestrial animals that depend on the vegetation in the treated water bodies. 3. 

### Discussion ofRisk Quotients No direct effects on listed animal species arepredicted. No exposure to terrestrial plants was expected, so no terrestrialplant RQs were calculated. Even though no RQs were calculatedfor aquatic plants, it is assumedthey will be affected, sincethey are the target organisms to be controlled. 

### 4. Probit Dose Response Relationship Since no mortality was observed in the standard studies, no probit analysis was performed. 

### 5. Data Related to Under-representedTaxa The screening level assessment relies on 

### RQ calculationsthat use toxicity endpoints selected from the most sensitive species tested within broad taxonomic groups. There may be I situations in which additional effects data from one or more sources may suggest that a given suite of listed taxa may be more or less sensitivethan suggestedby the effects data used for 

### RQ I calculations. No such relationships were found in this analysis.

\ 6. Implications of Sublethal Effects The Level 

### I screening assessment normally relies on the acute mortality endpoint as well as a suite of sublethal responses to the chemical stressor. 

### EFED decided that, based on its mode of action, the low chronic toxicity of at least one of the dyes to mammals, low potential for exposure based on use pattern, and its history as a human food dye, Aquashade did not need chronic toxicity studies. Sublethal effects were not analyzed. 7. Indirect Effects Analysis This assessment indicates that Aquashade may impact listed invertebrates, fish and amphibians that depend upon submerged aquatic plants and algae. There is also a potential for indrect effects to listed terrestrial birds and mammals that occur near treated water bodies, and that depend on submerged aquatic vegetation or algae for food or other survival orreproductive processes. The likelihood of potential indirect effects has not been quantified or refined. Such refinement could incorporate information on specificuse locations relative to specific locations of listed species. Subsequent refinement may also include additional analyses on potential indirect effects to listed species. 

### 8. Critical Habitat Aquashade may impact invertebrates and fish that depend upon the target species (submerged aquatic plants and algae). The likelihood of potential indirect effects has not been quantified. Subsequent refinement may include additional analyses on potential indirect effects to listed species. 9. Co-occurrence Analysis The goal of the analysisfor co-location is to determine whether sites of pesticide use are geographically associatedwith known locations of listed species. Listed species potentially at risk include aquatic plants, and animalsthat depend on aquatic effects from indirect effects. Aquashade's use pattern makes it (potentially) possible that it will be used in every county in theUnited States: Therefore, EFED's database to find which counties harbor which listed species ("LOCATES") was not used. However, there are possible effects to 

### D. Description of Assumptions, Limitations, Uncertainties, Strength, and Data Gaps 1. Environmental fate 

### Data Source The environmental fate assessment is only at the qualitative level. Structure-activity edtimations 

### (EPIWIN) of physical and chemical

### properties/characteristics are uncertain. 

### EPIWIN is not suitable for estimating properties of salts of large anions, such as in acid dyes. Therefore, 

### 1 thts qualitative environmental fate assessmentwas primarilybased on the general,known behavior of dyes in the environment and not on the specific behavior of Acid Blue 9 and Acid Yellow 23. 

### Reaction kinetics and transformation products Although indirect photolysisWas identified as a major route of dissipation of the dyes in aquatic media, the identity of the photoproducts (andhence, their toxicity)is not known. The rate of phototransformation is expected to be a function of geographical location and season. Biotransformationunder anaerobic conditions may also contribute to the dissipation of the dyes and can occur predominately in sediment,but the chemical identity of the metabolites is not known. 

### Impurities The characterization of the physical and chemical properties required under FIFRA were not' done with a 100% pure dye. The Acid Blue 9 is reported to be 50% pure and Acid Yellow 23 as being 28%. Although the presence of chemical impurities affect the physical and chemical properties of a chemical substance, the extent by which they affect those of the dyes is not known. T~c physical and chemical properties were submitted in support of Aquashade (23.6% Acid Blue 9 ahd 2.39 Acid Yellow 2.39% registration In is unclear if these percentages take into account the pu4ty of each dye. Furthermore, the percent purity of each dye in the other products is unknown. Given the low exposure of these dyes in the environment, when used aspesticides, no Subdivision

### N guideline studies are needed for the intended uses. However, if new uses are petitioned, the 

### EFED will reevaluate the status of data requirements. 2 Ecological toxicity The assessment assumes that Aquashade and related products will be used strictly in accordancewith label instructions. It assumes that there will be no large spills of the product I The ecological toxicity studies were not conducted with the individual dyes, but with the mixture of the two dyes as the test substance (23.6% Acid Blue 9 and 2.36 % Acid Yellow 23, AnAquashade end-use product). There are two other products that contain a higher percent of the dyes than the tested substance, namely Admiral Liquid (1 5.3

### 1 % Acid Blue 

### 9 and 1.00% Acid Yellow 23) and Admiral

### WSP (49. % Acid Blue 

### 9 and % Acid Yellow 23). Therefore, the ecological toxicity of these products is not known. However, the maximum estimated environmental concentration resulting from direct application to a water body is below the Level of Concern. The chronic toxicity RQs cannot be calculated, because there are no studies on chronic risk with the pesticide. The low acute toxicity values, along with testing with the Acid Yellow dye 23 with mammals, and the mode of action of Aquashade and other products, suggest low chronic toxicity and 

### EFED has not required any chronic studies. The Aquashade product that was used in the "basic six" studies was labeled as containing 231.63 % Acid Blue 9 and 2.39% Acid Yellow 23. The study authors stated, "The test substance characterization provided by the sponsor indicated a purity of 

### 13.9%azure blue dye." Therefore the exact composition of the toxicant that was used in the studies is not certain. The use of the wdrd "azure" could describe color, but also the word "azure" could be associated with a series of structurally related dyes that are totally different from Acid Blue 9 or Acid Yellow 23.V. Literature Cited E. OpenLiterature Books Lynch, D.G. "Estimating the Properties of SyntheticOrganic Dyes", in Handbook of Property I Estimation Methods for Chemicals- Environmental Health Sciences, Editedby Robert S. Boethling and Donald Mackay. Published by Lewis Publishers,Boca Raton, 

### FL, 2000; Pages I 447- 467. And pertinent references therein. Marmion,D,M. 

### Handbook of U.S. Colorants- Food, Drugs, and Medical Devices, Third Edition., 1991. Published 

### by John Wiley and Sons,'New York Helz, G.R., Zepp, R.G., and Crosby,D.G, Editors. 1994.Aquatic and surfacephotochemistry. Lewis Publishers,Boca Raton, 

### Florida.

Journal Articles Jank. M., Kijser, H., Liicking, 

### F., Martienssen, M., and Wittchen, S. 1998: 

### "Decolorization and Degradation ofErioglaucine (AcidBlue 9)Dye in Wastewater",Environmental Technology, ~.19(7), pp.741-747. Weber, E.J. and Adarns, R.L. 1995. 

### "Chemical and Sediment-Mediated Reduction oftheAzo Dye Disperse Blue 79",Environ. Sci. Technol. v.29,pp. 1163-1 170. Baughman, G.L. 1995. 

### "Fate of azo dyes in aquatic systems. Part 3: The role ofsuspended sediments in adsorption and reaction ofacid and direct dyes", Dyes and Pigments,v.27, pp. 197- 210. Brown, 

### D. and Laboureur,

### P. 1983, 

### "TheDegradation ofDyestufSs: Part 

### I- Primary biodegradation under anaerobic conditions 

### ", Chemosphere, v. 122,pp. 397-404. Baran,W.,Makowski, 

### A.,Wardas, W. 2003. 

### "The influence ofFeC13on the photocatalytic degradation ofdissolved azo dyes in aqueous Ti02suspensions", Chemosphere, v.53, pp 

### 82-95. Tai, W.T.,Chang, C.Y, Ing, C.H., and Chang,C.F. 2004. 

### "Adsorption of acid dyes from aqueous solutions on activated bleaching earth ", J. Colloid and Interface Science, vol. 275, pp 72-78. Mon, J., Flury, M., and Harsh, J.B. 2005


### ."Sorptionof four triarylmethane dyes in a sandy soil determined by batch and column experiments

### ",Geoderma. in press. I 1 Mon, J., Flury


### .M., and Harsh. 2005 "A

### quantitative structure-activity relationship analysis oftriarylmethane dye tracers ", Journal of Hydrology, in press. ,

I 'OSAR)Characterizationof Physical and Chemical Properties Submitted to the Agency in Support of Aquashade (23.6% and 2.39% Acid Yellow 23: Ellison. F.E. 1994. "Aqua Shade Blue 9'- Physical and Chemical Characteristicsof Aqua Shade Blue 9: Color, Physical State, Odor, Boiling Point, Specific Gravity, Solubility,Vapor Pressure, pH, and Stability". Performedby Case Consulting Laboratories, Inc. Whippany, NJ and sponsoredby Applied Biochemistry,Inc., Milwaukee, WI. Report Dated September 12, 1994. 

### MRID 43503401

I Ellison. F.E. 1994. "Aqua Shade Yellow 23"- Physical and Chemical Characteristics of Aqua shadeBlue 9: Color, Physical State, Odor, Boiling Point, Specific Gravity, Solubility,Vapor Pressure,pH, and Stability". Performed by Case ConsultingLaboratories, Inc. Whippany, 

### NJ and I sp'onsoredby Applied Biochemistry, Inc., Milwaukee, WI. Report Dated September 12, 1994. 

### MRID 43503402
5. Data Generation Structure-activity Estimates: EPI (Estimation Programs Interface) suiteTM (formerly known as EPIWIN): 

### htt1p://www.epa.govlopptintr/exposure/docs/episuite.htm SRC. 2004. SyracuseResearch Corporation. Interactive Physical Properties (PHYSPROP) Database Demo. Search terms: FD&C Blue. No. 1,FD&C Red No. 40, FD&C Yellow No. 

### 5. (November 28,2004) http://www.syrres.corn/esc/physdemo.htm

E. U.S. Environmental Protection Agency Documents: "Robust Summaries & Test Plans: C.I. Acid Yellow 23 (FD&C Yellow 5)", Submitted ti the Agency by the International Association of Color Manufacturers on March 10,2004. 

### http://www.epa.gov/chemrtWciacdylo/c15133tc.htm "Background Document for Identificationand Listing of Deferred Dye and Pigment Wastes, 

### U.S. EnvironmentalProtection Agency, Office of Solid Waste, Washington, D.C., June 1999. US EnvironmentalProtection Agency. 1998. Guidelines for Ecological Risk Assessment. Risk Ass'tssmentForum.EPN630lR-95/002F, April 1998 U.S


### .EPA. 1993. 

### U. S


### .EnvironmentalProtection Agency. Wildlife Exposure FactorsHandbook. Volume I of 11. EPN6OOlR-931187a. Office of Research and Development, 20460.

E. U.S. Environmental Protection Agency, Office of Pesticide Programs Documents:

IMemorandum: "Review of Phase 4 List D Package for Aquashade" (EFGWB # 93-0119, 93-0120; Chemical # 110301and 110302;Case # 819437 and 819438; DP Barcode Dl84289 and Dl84278).Dated February 4,1993. "Reassessment of the Exemption from the Requirement of a Tolerance for the FDACertified Color Additives FD&C Blue No. 1,FD&C Red No. 40, and FD&C Yellow No. 

### 5 (Tartrazine)". 

### U.S. Environmental Protection Agency, Office of PesticidesPrograms. December 2 1,2004 D. InternetResources Food and Drugs Administration: 

### http://www.cfsan.fda.gov/-dmslopa-col2.html Color Index International: 

### http://www.colour-index.org/ Additional informationwas also obtained from TOXNET. http://toxnet.nlm.nih.gov/

### Appendix A- Data Gaps EnvironmentalFate All of the Subdivision N, Environmental Fate Data Requirements maybe waived because ofvery low risk potential. They were identifiedas 'Reserved in 1993 (Memorandum: "Review of Phase 4 List D Package for Aquashade"; EFGWB#93-0119,93-0120; Chemicals 

### 11030 1 and 1 10302; Case # 

### 8 19437 and 8 19438;DP Barcode Dl84289 and Dl84278(. Dated February 4, 

### 1993),pending on the results ofrequired ecological toxicity studies. There are two major uncertainties on the environmental fate behavior ofAcid Blue 9 and Acid Yellow 23 : 

### (1) Structure-activity estimatesofphysical, chemical, and environmental fate behavior are not reliable for salts of large anions, such as Acid Yellow 23 and Acid Blue 9. (2) The major route oftransformation of dyes such as Acid Blue 9 

### and Acid Yellow 23 is indirect photolysis. However, the chemical identity of the photoproducts of these two dyes are not known. Biotransfomationunder anaerobic conditions may also contribute to the dissipation of the dyes in water-sediments. The chemical identity of the metabolites is not known. However, after the review of the ecological toxicity studiesand the risk assessment, the environmental fate datarequirements placed in 1993 as "Reserved' can be waived. The. Environmental Fate and EffectsDivision is not requiring any new environmental fate data for the two dyes individually or for any ofthe end-useproducts (Aquashade, Aquashade OA, Pond Care AlgaeBlocker, Admiral Liquid, and Admiral WSP). Ecological Toxicity b. Effects The only EFED ecological toxicity data available is for the Aquashade product (23.6% Acid Blue 9 and 2.36% Acid Yellow 23) as the test substance. There are no EFED ecological toxicity data for Admiral Liquid or Admiral WSP, but HED received a study that found a laboratoryrat LD50 equal to or more than 5,000 mgkg. However, since organismswill be only exposed to either 1or 

### 2 ppm in the environment, this uncertainty is minor. No chronic studies have been required, so it isnot known whatwould happen over an extended exposureperiod. No studies on aquaticplants have been required, because they are the targets ofAquashade and it is assumed that all sudmerged aquatic plants in the treated pond will be killed. No terrestrial plant studiesha+e been required, because there is little expected exposure. 

### An exception would be if anj. 

### ofthese productsmight be applied to water used for irrigation. If that occurred, exposure to terrestrial plants may occur. It is not known what would happen to plants on the margins of a treated pond or growing as emergent vegetation in the shallows of the pond if the technical product or the end-use product was applied to them. Some labels warn that directly applyingthe products to emerged orterrestrial plants may cause burning. Because of the mode of action (blocking light that enters bodies of water), the low acute toxicity of this pesticide, and the relatively limited area involved in treatment,it is believed that additional studies are not needed.

### Appendix B 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. Although the Environmental Fate and Effects Division does not usually use open literature data in its assessments,published data on the chemistry and behavior of dyes in aquatic systems was used to supplement the environmental fate assessment.Most of the data comes 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. The environmental fate behavior of dyes such as Acid Yellow 23 and Acid Blue 9 can be summarized as follows: The dyes are fully ionized in the environmentally significant pH range of 

### 5 to 

### 9. 1. There no hydrolyzable groups in these two dyes and, therefore, abiotic hydrolysis is not an important degradationpathway in the environment. 2. Direct photolysis is slow, as indicatedby the fading of the dyes in homogeneous aqueous media exposed to sunlight. However, indirect photolysis has been identified as the major route of degradationin environmental media. 3. The dyes are resistant to biodegradation under aerobic conditions. Structural related dyes are difficult to biodegrade in activated sludge. However, redox reactions under anaerobic conditions appear to degrade the dyqs, at least to some extent.
4. The two dyes are highly hydrophilic and do not have the potential to

' bioaccumulate in fish.

### Appendix 

### C Aquatic Exposure and Modeling The end-use products containing Acid Blue 9 and Acid Yellow 23 are directly applied to a confined water body and at the dose specifiedin the labels to attain a concentration of 1 ppm or 2 ppm depending on the target pest. Because there are no runoff or spray drift components, the use of the simulationmodels GENEEC orPRZM and EXAMS is not appropriate. Aquatic exposure was based on the target concentration specifiedin the product labels, depending on the weed to be controlled. The maximum target concentration is 2 ppm (2 ma) which is the concentration used to estimate risk for both aquatic and terrestrial animals

### Appendix 

### D. Ecological Toxicity and Uncertaintyin Ecological Toxicity Data
I EPED requires the submission of six studies for all chemicalsthat are submitted for registration. Which additional data that isrequired depends upon the circumstances of the chemical and its use pattern. Birds are used as surrogates for terrestrial phase

. amphibians and reptiles. Freshwater fish are used as surrogates for aquatic phase amphibians. The end-use products are not applied to a terrestrial environment. However, terrestrial animals (e.g.,mammals and birds) are exposed to the dyes when they use treated water as the source of drinking water. In addition to data that was submittedby the registrants,EFED alsoreviewed studies from the open literature. These studieswere obtained through the ORD Middle
I Ecological Division's ECOTOX literature search and retrieval program managed in Duluth, Minnesota 

### (http://www.epa.gov/ecotox). These data are consistentwith the I contention that the dyes found in Aquashade and the other end-useproducts are practicallynontoxic to the organisms tested.

Bobwhite quail Acute Avian Oral 433367-01 LD~o & NOAEL z 2250 x 0.13 292.5 mglkg Mallard duck Acute Avian Oral 433367-02 LD50 & NOAEL z 2250 x 0.13 r 292.5 mglkg Bobwhite quail Avian Dietary 435034-03 LC50 & NOAEL5 2 620 x 0.13 2 80.6 ppm Mallard duck Avian Dietary 435034-04 ' LC50 & NOAELS 2 620x 0.13 r 80.6ppm Bluegill sunfish Acute Toxicity 432975-02 LC50 & NOAEC r 96x 0.13 2 12.48mg/L I Rainbow trout Acute Toxicity 432975-01 LC50 & NOAEC t 96 x 0.13 t 12.48mgLL Daphniamagna Acute Toxicity 432975-03 I LC50 &NOAEC 2 97 x 0.13 2 12.61mg& I 1 All original tests were performed with the TypicalEnd-useProduct, Aquashade. No testing was performed on birds, fish and invertebrateswith the end-useproducts Admiral Liquid and Admiral WSP, which have ahigher percentage of the dyes than the tested Aquashadeproduct.

In addition to the uncertainty about the possiblepercent purity adjustment, Admiral WSP (49.27% acid blue 9 and 3.27% acid yellow 23) has about 50%greaterconcentration of dyes than Aquashade. Totake this higher concentration into account, each toxicity endpoint could be multipliedby 2 resulting in an extrapolatedavian acute N,OAEL 2 146 mg/kg and aquatic organism

### NOAEC 26.3 mg/L.71-la 1433367-01 Acute oral bobwhite quail 

# '
71-la/ 433367-02 Acute oral mallard duck 

# ' The formulated product (23.6% acid blue and LDS0 2 2250 mg eup/kg 2.39% acid yellow dyes) was used. The study concentration was not adjusted for the purity of NOAEC = the active ingredients (the two dyes). The 2250 mg eup/kg results are milligrams of the end-useproduct. Five experimental doses were used - 292,486, 810, 1350,2250mg/L. There were no mortalities or other adverse effects. 71-2b 1 435034-03 Acute dietary bobwhite quail 

# ' 7 1-2a/ 435034-04 Acute dietarywaterfowlmallard duck 

# ' - - The formulated product (23.6% acid blue and LC5,, eup 2 5,620ppm 2.39% acid yellow dyes) was used. The study concentration was not adjusted for the purity of NOAEC eup = the active ingredients, (the two dyes). The 5,620ppm eup results are parts per million of the end-use product. Five experirinental doses were used - 562, 1000, 1780,316b,and 5620ppm. There were no mortalities or other adverse effects. 71-3/ 452811-01 The study used Admiral WSP, a purple powder Acute Wild Mammal formulation. The dye,swere not otherwise Toxicityusing the identified. A group of 7-10 week old rats Laboratory rat (51sex)received a singledose of 5000 mgkg by gavage and were observed for 14days. There were no mortalities, therefore the chemicalwas categoired as 'Toxicity Category 

### IV." - -- Oral LDSO a a o >5,000mg eup/kg OralLD50 o' & 0 s2,500mg a.i. /kg

83-4 I 434109-01 Three generations ofLong-Evans rats were NOAEL.&e = Reproductionstudy given tartrazine (92% a.i.) at 7.5,75, 225, and 750 mglkglday using the laboratory 750 mglkglday in their diet. This is consistent rat with no effects and addresses whether acid yellow 23 causes reproductive toxicity. There were no adverse treatment related effects. 72-lc I 432975-01 The formulated product (23.6% acid blue 

### LC5o eup 2 96 

### ma Acute coldwaterfish- and 2.39% acid yellow dyes) was placed in 

### NOAEC eup 

### = Rainbow trout 

# ' the water of aquaria. The study 96mg/L 72-la 1 432975-02 concentration was not adjusted for the Acute warmwater purity of the active ingredients (the two fish- bluegill dyes). Theresults are milligrams of the end-use product. ThB only dose used was sunfish 

# ' I 1 96

### ma. I72-2a / 432975-03 The formulatedproduct (23.6% acid blue LC5()eup 2 97

### mg/L Freshwater and 2.39% acid yellow dyes) was placed in 

### NOAEC eup 

### = invertebrateDaphnia the water of aquaria. The study 1 97mg/L magna concentrationwas not adjusted for the purity of the active ingredients (the two dyes). The results are milligrams of the end-use product. The only dose used was 

### 97 

### mg/L. 72-1 a Freshwater cladocerans (Ceriodaphnia ECS0 = 5707mg/L Warne & Schifko, dubia) were exposed to a dye for two days. 1999 The EC50put the chemical in the category 

### FW Cladoceran "practically nontoxic." 72-3a Exposureto marinehabitats not expected N/A EstuarineIMarine Acute Toxicity with SheepsheadMinnow, Oyster, Mysids

, 72-4a Exposureto marinehabitats not expected N/A Freshwater Fish Early Life StageFathead minnow 72-4b Chronic effectsnot expected based on mode of N/A Freshwater Invertebrate Life action and lack of effects in acute tests. Cycle- Water flea 123-la If the labelis changed and the water is not used Reserved Tier I1 Terrestrial Plant for irrigation, exposure to terrestrial plants Seedling Emergence would be unlikely. (GF 871) 123-lb If the label is changed and the water isnot used Reserved Tier I1TerrestrialPlant for irrigation, exposure to terrestrial plants Vegetative Vigor (GF would be unlikely. 871) 123-2 Aquatic plants are the targets. Therefore, Tier I1 Aquatic Plant testing is not required. Release from Studies treated ponds to other water bodies considered a very low probability. ,141-1 Mode of action suggests low likelihood of NIA Honey Bee Acute toxicity. The method of application Contact Toxicity minimizes exposure to terrestrial insects. Nonguideline Crayfish were exposed to 3 levels of NOAEL = 15mg/L Spencer, 1984 Aquashade for 5 days. The oxygen use was Crayfish effected. The study was categorized as "slightly toxic." Nonguideline Laboratory rats were fed 4 levels of yellow 5 NOAEL = Borzelleca and for 900 days. The study was categorizedas 984 mg/kg/day Hallagan, 1988. "practically nontoxic." LOffiL = Chronictoxicity1 264 1mg/kg/day carcinogenicity

- - - --- Nonguideline House mice (Mus musculus) were given food 7354 mglkglday Borzelleca, et al., 1990 with 3 levels of dye in their diet for 720 days. Chronic toxicity1 The study was characterized as "practically carcinogenicityin nontoxic." mice

- --- I The study was done with Aquashade. The doses were calculated with the end-use product. The percentages of the dyes making up liquid is not clear.

The study was done with Admiral Lake Colorant

### WSP. The 

### LDsois based on the dry weight of the end-use product. The compositionof the powder was not determined,but it is usually 49.62% blue dye and 3.05% yellow dye. The 

### LDSO a.i. cannotbe determined,but is probably 50% of the stated >5,000 mglkg.

he study was done with FD&C yellow dye (acid yellow 23 or 5),which is not an end-use products. 

### Uncertainty in toxicity information Most of the toxicity results used for risk assessment were based on effects testing done with Aquashade (23.6% acid blue 

### 9 and 2.39% acid yellow 23) ekcept the acute mammal 

### LDSo was done with Admiral WSP and the reproductivemambalian 

### NOAEL was done with acid yellow. There is some uncertainty in this approach because 1)the percent of the dyes in this formulationis unclear, and 2) some formulations have a higher percent of each dye than Aquashade, and it is not clear if the doses were expressedin terms of end-use product or active ingredients. The dyes in Aquashade are reported as bking 

### 13%pure. To demonstrate the degree of uncertainty,the toxicity values may be mbltiplied by 0.1 to I

Itake these percentages into account. Note that the risk assessment was conducted with actual toxicity test results, not with these adjusted toxicity values.

Table F-1. Summary of acutetoxicity endpoints multipliedby 13%to compositefor the possibility of the tested materials being only 13%pure. species Study T~P~' Source ExtrapolatedEndpoint Bobwhite quail Acute Avian Oral 433367-0 1 LD & NOAEL > 2250 x 0.13 2 292.5 mg&g so Mallard duck AcuteAviao Oral 433367-02 LD & NOAEL 2 2250 x 0.13 2 292.5mgkg 50 Bobwhltequa11 Avian Dietary 435034-03 LC &NOAEL 2 620 x 0.13 2 80 6ppm 50 Mallard duck Avian Dietary 435034-04 LC &NOAEL 2 620 x 0.13 2 80.6 ppm 50 Bluegill sunfish Acute Toxlcity 432975-02 LC &NOAEC >96x0.13 >1248mgn 50 Rainbowtrout Acute Toxicity 432975-01 LC &NOAEC 2 96 x 0.13 2 1248 mgL 50 Daphnra magna Acute Toxicity 432975-03 LC &NOAEC 297~0.13 >1261mgL 50 AH ofiginaltests Were performed with the typical end-useproduct, Aquashade. No testing was performedon bnds, fish and invertebrates with the end-use products AdmiralLiquid and AdmiralWSP, which have a higher percentageof the dyesbmthe tested Aquashade product. In addition to the uncertainty about the possible percent purity adjustment, Admiral 

### WSP (49.27% acid blue 9 and 3.27% acid yellow 23) has about 

### 50%greater concentration of dyes than Aquashade. To take this higher concentration into account, each toxicity endpoint could be multiplied by 2 resulting in an extrapolated avian acute NOAEL 2 146mglkg and aquatic organismNOAEC 26.3 mgL.

### Appendix 

### E Status

### ofData Requirements 1. Environmental Fate Subdivision N, Environmental Fate Data Requirements- Status 

### of Data Requirements 

# 11 161-1 [Abiotic]Hydrolysis Reserved May be Waived No I I I Data Requirement

162-1 Aerobic Soil Metabolism Not Required Not Required No Status in 1993

161-2 [Direct] Photolysisin Water 161-3Photolysis on Soil 

# 11 162-2Anaerobic Aquatic Metabolism 

# INot Required 

# I Not Required 

# INo Status after the Risk Assessmentfor Reregistration

Reserved
Not Required Additional Data Required

162-3Anaerobic Aquatic Metabolism 

# 11 164-1Terrestrial Field Dissipation 

# INot ~e~uired Not Required No I I I May be Waived
Not Required

162-4Aerobic AquaticMetabolism 163-1 Mobility in SoilsISediments 

# 11 164-2Aquatic Field Dissipation Reserved May be Waived No No No

I I

Reserved Reserved Reserved

Acute Avian Oral Quail TEP Y 433367-01 9-7-94 Acceptable 7 1-1(b) Acute Avian Oral Duck TEP Y 433367-02 9-7-94 Acceptable 7 1-2(b) Avian DietaryfQuail TEP Y 435034-03 4-26-95 Acceptable 7 1-2(b) Avian DietaryDuck TEP Y 435034-04 4-26-95 Acceptable 71-3 Wild Mammal Toxicitya Y 4528 11-01 12-5-00 Acceptable 7 1-4(a) Avian ReproductiveIQuail Waived 7 1-4(b) Avian ReproductivetDuck Waived 72-l(b) Acute Toxicity Bluegill TEP Y 432975-02 4-25-95 Acceptable 72-l(d) Acute Toxicity Rainbow Trout TEP Y 432975-01 4-25-95 Acceptable - ---- May be Waived

165-4Bioaccumulation in Fish -- No May be Waived May be Waived No No

I Reserved May be Waived The Log Kow does not triggerthis data requirement No- This study was performed with AdmiralWSP No additional effects data are being required.Appendix 

### F Environmental Fate Bibliography Books Lynch, D.G. "Estimating the Properties of SyntheticOrganic Dyes", in Handbook of Property Estimation Methods for Chemicals-Environmental Health Sciences, Edited by Robert S. Boethling and Donald Mackay. Published by Lewis Publishers, Boca Raton, 

### FL; Pages 447- 467. And pertinent references therein. Marmion,D,M. Handbook of 

### U.S. Colorants- Food, 

### Drugs, and Medical Devices, Third Edition., 1991. Published by John Wiley and Sons, New York Helz, G.R., Zepp, R.G., and Crosby, D.G, Editors. 1994.Aquatic and surface photochemistry. Lewis Publishers,Boca Raton,Florida.

~ourn'al Articles Jank. M., Koser, 

### H., Liicking,

### F., Martienssen, M., and Wittchen, S. 1998, 

### "Decolorizationand Degradation ofErioglaucine (AcidBlue 9)Dye in Wastewater," Environmental Technology,v. 19(7),pp.74 1-747. Weber, E.J. and Adams, R.L. 1995. 

### "Chemical and Sediment-Mediated Reduction ofthe Azo Dye Disperse Blue 

### 79,"Environ. Sci. Technol. v.29, pp. 1163-1 170.

I Baughman, G.L. 1995. 

### "Fate ofazo dyes in aquatic systems. Part 3: The role of suspended sediments in adsorption and reaction ofacid and direct dyes," Dyes and Pigments, v.27, pp. 197-210. Brown, D. and Laboureur,

### P. 1983, 

### "TheDegradation ofDyestuffs: Part 

### I- Primary biodegradation under anaerobic conditions, " Chemosphere, v. 122,pp. 397-404. Baran,W.,Makowski,A., Wardas, W. 2003. 

### "The influence ofFeCl3 on the photocatalytic degradation ofdissolved azo dyes in aqueous Ti02suspensions, " Chemosphere, v.53,pp 82-95. Tai, W.T., Chang, C.Y, Ing, 

### C.H., and Chang,C.F. 2004. 

### "Adsorptionofacid dyes from aqueous solutions on activated bleaching earth, " J. Colloid and Interface Science, vol. 275, pp 72-78. Mon, J., Flury, M., and Harsh, J.B. 2005 

## ."Sorption of four triarylmethane dyes in a sandy soil determined by batch and column experiments, " Geoderrna.in press.

I I Mon, J., Flury. M., and Harsh. 2005 "A quantitative structure-activity relationship (OSAR) analysisof triarylmethanedye tracers," Journal of Hydrology, in press.

\Appendix 

### G Ecological Toxicity Bibliography

### Appendix 

### H. Endangered and Threatened Species EFED also assessed the potential for Aquashade and other end-use products containing the Acid Yellow 23 and Acid Blue 

### 9 dyes to potentially cause harm to endangered species. 

### Terrestrial Animals Listed endangered or threatened species of birds, mammals and reptiles are not I I expected to be affected directly because exposure does not exceed the endangered species LOC. However, terrestrial animals may be affected indirectly if aquatic plants that are necessary for their survival are eliminated. The degree to which this might occur depends on the extent the treated water bodies are a critical component of their food supply. 

### Terrestrial Plants Terrestrial plants have the potential for exposure if water treated with Aquashade or the other dye containing end-use products are used for irrigation. If irrigationis precluded fromuse on the labels, 

### EFED concludes there is virtuallyno potential for exposure to terrestrial plants. 

### Aquatic Animals Listed species of fish, invertebrates or amphibians are not expected to be affected directly because the exposure does not exceed the endangered species LOC. However, aquatic animals dwelling in treated ponds might have the potential for harm if the plants on which they depend for food and habitat are eliminated. The likelihood of this occurring depends on if any listed aquatic animal species would occur ,in these highly managed, often manmade, normally relatively small water bodies with usually no outflow, or if any, a very small amount. 

### Aquatic Plants Listed species of aquatic plants which begin their life stages as submergedmight be harmed if water bodies in which they are growing are treated while they are submerged. 

### EFED has not fully analyzed the potential for endangered specits to be harmed through use of Aquashade and other end-use products. Such analysis a/ould focus on the overlap of specific species with potentially treated water bodies, and inlthe case of indirect effects, the likelihood that the treated water body comprises enbugh of a component of the listed species food or habitat base to actually impact the listed species, if that food or habitat was lost.

### Appendix 

### K. Risk Quotient Calculation and Interpretation Risk characterization integrates the results of the exposure and ecotoxicity data to evaluate the likelihood of adverse ecological effects. The means of this integration is called the quotient method. Risk quotients (RQs) are calculatedby dividing exposure estimatesby acute and chronic ecotoxicity value and, in this case,it is a deterministic approach. RQ = EXPOSURE/TOXICITY RQs are then comparedto OPP's levels of concern (LOCs). These LOCs are used by OPP to analyze potential risk to nontarget organisms and the need to consider regulatory action. The criteria indicate that a pesticide used as directed has the potential to cause adverse effects on nontarget organisms. LOCs currently address the following risk presumption categories: (1)

### acute -- potential for acute risk; regulatory action may be warranted in addition to restricted use classification, (2) 

### acute restricted use -- the potential for acute risk, but may be mitigated through restricted use classification, (3) 

### acute endangered species - endangered species may be adversely affected, and (4) 

### chronic risk - the potential for chronic risk; regulatory action may be warranted. Currently,EFED does not perform assessmentsfor chronic risk to plants, acute or chronic risks to nontarget insects, or chronic risk from granularlbaitformulations to birds or mammals. The ecotoxicity test values (measurement endpoints)used in the acute and chronic risk quotients are derived from required studies. Examples of ecotoxicity values derived from short-term laboratory studies that assess acute effects are: (1) 

### LCso(fish and birds), (2) 

### LDSo (birds and mammals), (3) ECso(aquatic plants and aquatic invertebrates) and (4) EC25 (terrestrial plants). Examples of toxicity test effect levels derived from the results of long-term laboratory studies that assess chronic effects are: (1) LOAEC (birds, fish, and aquatic invertebrates), and (2) NOAEC (birds, fish and aquatic invertebrates).For birds and mammals, the NOAEC generallyis used as the ecotoxicity test value in assessing chronic effects, although other values may be used whenjustified. However, the NOAEC is used if the measurement end point is production of offspring or survival

### Risk presumptions and the correspondingRQs and LOCs, are tabulated below.
Table K-1: 

### Risk presumptions for terrestrial animals Risk Presumption RQ LOC Birds Acute High Risk EEC'/LCSOor ~~50ls~ft~ or ~~50lda~' 0.5 Acute RestrictedUse EECILCSO or LDSOlsqft orLD5OIday (or LD50 

### <50mglkg) 0.2 Acute Endangered Species EECILCSO or LDSOlsqft or LDSOIday 0.1 Chronic Risk EECINOAEC 1 I Wild Mammals

I Acute High Risk EECILCSO or LD5OIsqft or LD5OIday 0.5 Acute Restricted Use EECILCSO or LD501sqft orLD5OIday (or LD50 

### < 50mglkg) 0.2 Acute Endangered Species EECILCSO or LDSOlsqft orLD5OIday 0.1 Chronic Risk EECINOAEC 1

1 abbreviation for EstimatedEnvironmental Concentration 

### (ppm) on avianlmammalian food items 2 mg/ft2

mg of toxicant consumed/day 

### LD50 

## * wt. of a birdTable 

### K-2.Risk Presumptionsfor Aquatic Animals Risk Presumption RQ LOC Acute Risk EEC'LCSO or EC50 0.5 Acute Restricted Use EECLCSO or EC50 0.1 Acute Endangered Species EECLC5O or EC50 0.05 Chronic Risk EECIMATC or NOAEC 1
1 EEC = (ppm 

### or ppb) in water

Table K-3. Risk presumptions for plants Risk Presumption RQ LOC Terrestrialand Semi-Aquatic Plants Acute Risk EEC'/EC~~ 1

Acute High Risk EEC'/ECSO 1
1 Acute Endangered Species EECECO5 or NOAEC 1

1 2 EEC = lbs ai1A EEC = (ppblppm) in water