9/10/13 L;-.I I TED STATES EI'I'V IR O"'\ 1 EI\TAL PROTECT I O N AGE NC Y
WAS HI NGTON, D

. C. 20460

OFF I C E O F C HEMI CA L SA F E l Y

AN I> PO L L U TIO N PR EVEN TIO N

MEMORANDUM

SUBJECT: Petition #2E8004; PC code 911325 ; FD&C Blue No. 1 (CAS Reg. No. 3844-45-9); Human Health Risk Assessment and Ecolog ical Effects

Assessme nt to Support Proposed Exemption from the Requirement of a Tolerance When Used as an Inert Ingredient in Pesti cide Formulations.

All PC Code: 911325

Decision No.: N/A DP Barcode: N/A Regulatory Action: Inert

Tolerance Exemption; 40

CFR 180.920

Petition No: 2E8004 All CAS N o: 3844-45-9

Inert Tracking No: N/A

FROM: E lizabeth Fertich 

# ..--[i___(}

Inert I ngredient Assessment Branch ( II AB) 

# £.**~** 

# o/toj t3

Registration Division (7505P)

TO: PV Shah

, C hief

Inert Ingredient Assessment Branch (IIAB) Regis trat ion Divi sion (7505P)

EXECUTIVE SUMMARY

This is the dec isio n document for FD&C Blue No . 1 (CAS Reg. No. 3844-45-9). The firm Exponent ( 1150 Connecticut Ave.

, NW; Suite 11 00 ; Washington

, DC 20036)

, on behalf of

Sensient Colors

, LLC (2515 N. Jeff erson Ave.; St. Loui s, MO 63106) has submitted a request to

the Environmental Prot ection Agency (E P A or " the Agency") to exempt from the requirement of a tolerance FD&C Blue No. 1 when used as a see d treatment/dye under 49 CFR 180.920. The

chemical i s aJso referred to as Brilliant Blue FC F in thi s document

, as this name is synonymous with F D&C Blue No

. l.

12 FD&C Blue No. 1 is currently approved for use as an inert ing redient in pesticide products under 40 CFR 180.910 as a dye in pre

- and post

- harvest applications, at a maximum of 0.2% by weight in final pesticide formulations and under 40 CFR 180.930 as a dye, and coloring agent in applications to animals. It is also approved for use as an active ingredient as an aquatic plant control agent, under 40 CFR 180.1074. The primary use pattern of registered products containing this active ingredient is as a pre

- emergent aquatic plant control agent. The products are design ed to absorb specific wavelengths of light cr itical to plant photosynthesis and are used in natural or manmade contained lakes and ponds with little or no outflow of water, golf course ponds and watering turf. In addition to pesticide uses, t he Food and D rug Administration (FDA) and Joint FAO/WHO Expert Committee on Food Additives (JECFA) have assigned maximum acceptable daily intakes (ADIs) for FD&C Blue No. 1 and it is widely used as an ingredient in food products and personal care products.

FD& C Blue No. 1 is not acutely toxic via the oral route in rats and via subcutaneous injection in mice given the median lethal doses (LD50) were greater than 2000 mg/kg body weight and 4600 mg/kg body weight, respectively. There was also no evidence of sub

- c hronic toxicity is a study with mice fed 1200 mg of Brilliant Blue over a time period of 19 days.

In a chronic dog study, beagle dogs fed 1% or 2% (equivalent to 1000 or 2000 mg/kg bw/day) for a time period of one year showed no treatment related clinica l signs of toxicity or histological

abnormalities.

No evidence of carcinogenic action was found in study with mice fed 1 mg /day in the diet for 500

- 700 days. Similar results were found in a study with rats fed 4% (equivalent to 2000 mg/kg bw/day ) for 6 00 days. There was also no evidence of tumors in 85 rats fed 0.1% of the blue dye for their entire lifespan.

Group of 24 weanling Osborne

- Mendel rats (12 males and 12 females) were fed the color at levels of 0, 0.5%, 1.0%, 2.0% and 5.0% (equivalent to 250, 500, 1000 or 2500 mg/kg bw/day) for a time period of 2 years. Gross and microscopic examination revealed incidental lesions, unrelated to treatment, with no effect on tumor production. The statistical evaluation revealed no changes in organ weights, mortality or growth and hematology was normal. No evidence of treatment

- related neoplastic lesions were observed in this study T he no observed adverse effect level ( NOAEL ) for this study is 5% ( equivalent to 2500 mg/kg bw/day) Brilliant Blue FCF in the d iet, the highest dose

tested.

Wistar rats were fed diets containing 0, 0.03%, 0.3% and 3.0% (equivalent to 15, 150 or 1500 mg/kg body weight (bw )/day) of the color for 75 weeks. No effects on growth, food consumption, and food efficiency were found. Mortality was increased in the 3% group, however this was determined to be a result of respiratory infection unrelated to administration of the test substance in the diet. Hematology findings were normal. The NOAEL for this study is 1500 mg/kg bw/day, the highest dose tested.

Brilliant Blue FCF was administered in the diet to 360 mice at dietary levels of 0.5, 1.5 or 5% (equivalent to 661, 2064 and 7354 mg/kg bw/day in males and 819, 2562 and 8966 mg/kg bw/day3 in females). Lifetime exposure of mice to Bril liant Blue FCF as part of the diet did not result in consistent biologically significant, compound related adverse effects on behavior, morbidity, mortality, hematology, general physical observations or tumor incidence. The decrease in mean body weights of treated mice were thought to be a result of the non

- nutritive nature of Brilliant

Blue FCF. The NOAEL for this study was determined to be 7354 mg/kg bw/day for male and 8966 mg/kg bw/day for female, the highest doses tested.

Rats were treated with Brilliant Blue FCF in a chronic toxicity study coupled with a reproductive study. An in utero phase was conducted in which the test compound was admi nistered to the F0 generation. For the chronic phase, 700 rats were treated at the start of the study (70/sex /group, including 2 control groups). Following completion of the in utero phase, a maximum of 2 rats per sex from each litter were randomly selec ted for the chronic phase. The offspring were fed at dietary levels of 0.1, 1 or 2% Brilliant Blue FCF (equivalent to 50, 514 and 1073 mg/kg bw/day for males and 62, 631 and 1318 mg/kg bw/day for females). The treatment period was 116 and 111

weeks for m ale and female rats respectively. The NOAEL was equivalent to the dietary concentration of 2% (equivalent to 1072 mg/kg bw/day) for male rats and 1% (equivalent to 631 mg/kg bw/day) for females, based on a 15% decrease in terminal mean body weight and dec reased survival in high dose females.

Rats were treated with Brilliant Blue FCF in a chronic toxicity study coupled with a reproductive study. 60 males and 60 females from the F0 generation were fed at dietary levels of 0.1, 1 or 2% Brilliant Blue FCF (e quivalent to 50, 514 and 1073 mg/kg bw/day for males and 62, 631 and 1318 mg/kg bw/day for females) for 62 days. There were no compound related effects on fertility, gestation, parturition, lactation, pup survival through weaning, or on number of live and stillborn pups. One female in each test group died, however, these deaths were not considered treatment

related. The NOAEL is considered to be the dietary concentration of 2% (equivalent to 1073 mg/kg bw/day) for male rats and 2% (equivalent to 1318 mg/k g bw/day) for females.

Brilliant Blue FCF was fed to three successive generations of male and female rats at dose levels of 0, 10, 100, 300 or 1000 mg/kg bw/day. There were no treatment related effects on adult mortality, mating, pregnancy and fertility rates, lengths of gestation period, offspring survival or sex, litter survival or necropsy findings in animals sacrificed on day 19. There were also not macroscopic or microscopic tissue alterations for F1b and F3a generations that were attributable to

di etary administration of Brilliant Blue FCF. The NOAEL for this study was 1000 mg/kg bw/day, the highest dose tested.

Brilliant Blue FCF was fed to groups of mated Long Evans rats fro m day 6 to 15 of gestation at 0, 200, 600, and 2000 mg/kg bw /day. Administration of the color via the diet did not result in any fetal toxicity or anomalies. No further details were provided; therefore no NOAEL can be derived from this study.

Brilliant Blue FCF was administered orally to mated female New Zealan d White rabbits from day 6 to 18 of gestation at 0, 20, 60 or 200 mg/kg bw/day, administered in v o lumes of 1 ml/kg for all

treatment groups. Administration of the color via the diet did not result in any fetal toxicity or

anomalies. No further details we re provided; therefore no NOAEL can be derived from this study.4 Mice were given diets containing 0, 0.015, 0.15 pr 1.5% (equivalent to 0, 20, 200, and 2000 mg/kg bw/day) of Brilliant Blue FCF for up to 80 weeks. No consistent treatment related effects were found. Tumor pathology seen in treated mice, included squamous cell carcinoma of the stomach. A single occurrence of thyroid adenoma was found in the 30 mid

- dose males. Adrenal tumors were also found. A single occurrence of mesenteric lipoma was fo und in low dose males and squamous

cell carcinoma of the skin was found in 1 of 26 mid

- dose females and 1 of 39 high dose females. None of the tumor incidences in the treated animals were individually statistically significantly higher than the control. This study did not show any carcinogenic effects of Brilliant Blue FCF.

Five genotoxicity studies are available. Based on the results of four of these studies, it was concluded that Brilliant Blue FCF is not of concern with respect to genotoxicity. The results of the fifth study were determined to be inconclusive.

A study focused on synergistic interactions between commonly used food additives in an in vitro developmental neurotoxicity test using neuroblastoma cells. Developmental neurotoxicity was measured as an inhibition of neurite outgrowth in 24 hours. It was determined that Billiant Blue FCF (0.05

- 500 nM) in combination with L

- glutamic acid (0.5

- 100 μM) synergistically reduced neurite outgrowth length. In addition to the study referenced above, neurotoxicity was not observed in multiple, long

- term toxicity st udies.

Immunotoxicity studies were not available for review. However, signs of i mmunotoxicity were not observed in any of the available studies. EPA concluded that i mmunotoxicity studies are not required at this time.

In one study to determine the m etabolism potential of Brilliant Blue FCF, 14 C Brilliant Blue FCF

radiolabeled on 7

- C methylene position (sodium salt) was given by gavage at a dose of 1.5 mg/kg/bw to female Sprague

- Dawley rats. About 0.27% of the test substance was absorbed through the gastrointestinal tract. The major route of excretion was through the feces with total recoveries being 92%. In a second metabolism study, male and female rats (strain unspecified) were dosed by gavage with either 30 mcg/kg bw or 3 mg/kg bw of 14 C Brilliant Blue FCF. 99.9% of the lower dose was excreted in the feces within 72 hours, while 95.4% of the high dose was excreted in the feces in the same time period. Pregnant rats were given 3 mg Brilliant Bl ue FCF on day 8 of pregnancy. It was determined that practically all of th e administered dose was excreted in the feces and the urinary content of radioactivity was less than 1%

. The lack of gastrointestinal absorption and metabolism was confirmed by stu dies in guinea pigs and mice using isolated loops

of small intestines. Brilliant Blue FCF was administered orally to rats at 2% aqueous solution (200 mg per rat). In this test, almost the entire amount of the test substance was excreted unchanged in

the feces within 40 hours after administration.

The available toxicity stud ies indicate that FD&C Blue No. 1 has a very low overall toxicity. The NOAEL is 631 mg/kg bw/day. Since no endpoint of concern was identified for the acute and chronic dietary expos ure assessments and short and intermediate terms dermal and inhalation exposure assessments, a quantitative risk assessment for FD&C Blue No. 1 is not necessary.

There is no evidence of qualitative or quantitative susceptibility of infants and children i n the available database. As part of its qualitative assessment, the Agency did not use safety factors for assessing risk, and no additional safety factor is needed for assessing risk to infants and children.5 The available toxicity studies suggest low to xicity of FD&C Blue No. 1. The toxicity database for FD&C Blue No. 1 contains an acute oral toxicity study, sub

- chronic and chronic toxicity studies, including carcinogenicity, reproductive and developmental toxicity studies. The database also

contains mu tagenicity studies, neurotoxicity data and metabolism data. There is no indication, based upon the available data, that FD&C Blue No. 1is a neurotoxic or immunotoxic chemical or results in increased qualitative or quantitative susceptibility in infants or children. Based on this information, there is no concern, at this time, for increased sensitivity to infants and children to this chemical when used as inert ingredient in pesticides formulations.

Dietary exposure to FD&C blue No. 1 can occur from eati ng food treated with pesticide formulations containing this inert ingredient. Non

- dietary exposure from the proposed use as a

seed treatment /dye under 40 CFR 180.920 is not expected, however non

- dietary exposure may occur from other approved uses of this chemical . Occupational exposure is also possible. However, the available data indicate that there are no adverse toxicological effects which occur at exposures less than 631 mg/kg bw/day, therefore a quantitative risk assessment is not conducted.

EPA ha s not found FD&C Blue No. 1 to share a common mechanism of toxicity with any other substances, and FD&C Blue No. 1 does do not appear to produce a toxic metabolite produced by other substances. For the purposes of this tolerance action, therefore, EPA has assumed that the

FD&C Blue No. 1 do es not have a common mechanism of toxicity with other substances.

In terms of terrestrial

, aquatic and atmospheric fate, Brilliant Blue is expected to be mobile in soil as it is expected to exist almost entirely in anion form in the environment. A nions do not adsorb more strongly to soils containing organic carbon and clay than their neutral counterparts.

Volatilization from moist soil and water surfaces is not expected. Due to the ionic nature of this chemical, bioconcentration in aquatic organisms is not expected. Ecological studies indicate that toxic effects are not likely to occur.

P otential areas of environmental justice concerns, to the extent possible, were considered in this human health risk assessment, in accordance with U.S. Executive Order 12898, "Federal Actions to Address Environmental Justice in Minority Populations and Low

- Income Populat ions," http://www.eh.doe.gov/oepa/guidance/justice/eo12898.pdf

.

Based on the lack of toxicological effects of concern and upon the conditions discussed above

, the

EPA concludes that th ere is a reasonable certainty that no harm will result to the general population, or to infants and children from aggregate exposure to residues of the FD&C Blue No. 1 when used as an inert ingredient in pesticide formulations under 40 CFR 180. 920

.6 I

. BACKGROUND

This is the decision document for FD&C Blue No. 1 (CAS Reg. No. 3844

- 45

- 9). The firm Exponent (1150 Connecticut Ave., NW; Suite 1100; Washington, DC 20036), on behalf of Sensient Colors, LLC (2515 N. Jefferson Ave.; St. Louis, MO 63106) has submitted a request to the Environmental Protection Agency (EPA or “the Agency”) to exempt from the requirements of

a tolerance FD&C Blue No. 1 when used as a seed treatment /dye under 49 CFR 180.920. The

chemical is also referred to as Brilliant Blue FCF in this document, as this name is synonymous

with FD&C Blue No. 1.

Use of FD&C Blue No. 1 as an inert ingredient under 40 CFR 180.910 and 180.930 was evaluated in by the EPA in 2005 in the document entitled “Inert Ingredient Tolerance Reassessment

— FD&C

Bl ue No. 1, FD&C Red No. 40 and FD&C Yellow No. 5 (Tartrazine). Brilliant Blue FCF was evaluated by the Joint FAO/WHO Expert Committee on Food Additives (JECFA) in 1970 and the EU Scientific Committee for Food (SCF) in 1975

. The most recent assessment of B rilliant Blue

FCF was conducted by the European Food Safety Authority (EFSA) Panel on Food Additives and Nutrient Sources added to Food (ANS) in the document entitled, “Scientific Opinion of the Re

evaluation of Brilliant Blue FCF (E 133) as a Food Additi ve,” published in 2010. During this assessment, the EFSA panel re

- evaluated all of the existing data on Brilliant Blue FCF and chose NOAELs, where the data supported the establishment of NOAELs.

II . Inert Ingredient Profile

A. Summary of Uses

1. Proposed Use Pattern

The proposed use pattern requested in this petition for FD&C blue No. 1 is as a seed treatment dye in pesticide formulations. The submitter is requesting a tolerance exemption under 40 CFR 180.920 (applications pre

- harvest) for resi dues of FD&C blue No. 1 when used as an inert ingredient without limitation in final pesticide formulations.

2. Inert uses

FD&C blue No. 1 is currently approved for use as an inert ingredient in pesticide products under 40 CFR 180.910 as a dye in pre

- and p ost

- harvest applications, at a maximum of 0.2% by weight in final pesticide formulations. It is also approved under 40 CFR 180.930 as a dye, and coloring agent in applications to animals.

3. Use as an active ingredient in pesticide products

FD&C blue No. 1 is approved for use as an active ingredient as an aquatic plant control agent, under 40 CFR 180.1074. There are currently four registered pesticide products that contain this chemical as an active ingredient. There are also four products pending7 regi stration with the Agency. The primary use pattern of the registered products is as a pre

emergent (prior to plant germination) aquatic plant control agent. The products are designed to absorb specific wavelengths of light critical to plant photosynthesis and are used in natural or manmade contained lakes and ponds (ornamental, recreational, fish bearing or fish farming) with little or no outflow of water, golf course ponds and watering turf. The products may not be applied to streams or other natural bo dies of water, or any body of water not under control of the user. They also cannot be applied to water intended for human consumption. These products are applied prior to the growing season and may be applied directly to ice cover, where applicable. Th e products may be used during the growing season to suppress the rate and extent of underwater plant growth or algae bloom.

4. Other uses

The Food and Drug Adminstration (FDA) and Joint FAO/WHO Expert Committee on Food Additives (JECFA) have assigned maximum acceptable daily intakes (ADIs) for

FD&C Blue No. 1 and it is widely used as an ingredient in food products such as ice cream, bottled food coloring, icings, ice pops, dairy products, sweets and drinks. It is also widely

used in personal care products such as soaps, shampoos and mouthwash.

Table 1. FDA and JECFA Acceptable Daily Intake (ADI) Values (in mg/kg

– bw/day

Color Additive FDA ADI JECFA ADI

FD&C Blue No. 1 12.0 0

- 12.5

Table 2. FDA Approved Color Additive Uses

Chemical 21 CFR 74 Uses

FD&C Blue No. 1 74.101 Foods generally

74.1101 Ingrested drugs generally; Externally applied drugs; Eye

area use 74.2101 Cosmetics generally; Eye are

use8 III. Physical Chemical Properties

Some of the physical and chemical characteristics of FD&C Blue No. 1, along with its structure and nomenclature, are found in Table 3

.

Table 3

. Physical and Chemical Properties of FD&C Blue No. 1

Parameter Value Source

Structure ChemIDplus Lite, 2013

CAS Chemical Name Benzenemethanaminium, N

- ethyl

N

- (4

- ((4

- (ethyl((3

- sulfophenyl)methyl)amino)phenyl)(

2

- sulfophenyl)methylene)

- 2,5

- cyclohexadien

- 1

- ylidene)

- 3

- sulfo

- , hydroxide, inner salt,

disodium salt ChemIDplus Lite, 2013

Chemical Formula C37

- H36

- N2

- O9

- S3.2Na ChemIDplus Lite, 2013

Synonyms Brilliant Blue FCF , 1206 Blue, Acid Sky Blue A, Brilliant Blue E 133, C.I. Acid Blue 9, disodium salt,

Food Blue 1 ChemIDplus Lite, 2013

CAS # 3844

- 45

- 9 ChemIDplus Lite, 2013

SMILES Notation C(=C1 \ C=C \ C(=[N+]( \ Cc2cc(ccc2)S(=O)(= O)[O

- ])CC)C=C1)(/c1c(cccc1)S(=O)(=O)[O

- ])c1ccc(cc1)N(Cc1cc(ccc1)S(=O)(=O)[O

- ])CC.[Na+].[Na+] ChemIDplus Lite, 2013

Molecular Weight 792.859 SRC, 2004 Water Solubility 1374 mg/L @ 25 °C (E) EPI Suite, 2004 Melting Point (°C) >250 °C (M) BIBRA, 1999

Vapor Pressure 2.97 x 10 - 42 mm HG @ 25 °C (E) EPI Suite, 2004

Henry's Law Constant 7.6 x 10 - 34 atm

- m 3 /mole @ 25 °C (E) EPI Suite, 2004

Partition Coefficient (usually the Log P or Kow) Log K
ow =

- 0.320 (E) SRC, 20049 I V

. Hazard ASSESSMENT

A. Summary of Toxicity Data.

FD& C Blue No. 1 is not acutely toxic. Long term studies of the effects of the color administered in the diet to rats and mice did not indicate any significant toxic effects. One chronic toxicity study coupled with a reproductive study resulted in a NOAEL of the dietary concentration of 2% (equivalent to 1072 mg/kg bw/day) for male rats and 1% (equivalent to 631 mg/kg bw/day) for females, based on a 15% decrease in terminal mean body weight and decreased survival in high dose females. A long term carcionge ncity study did not show any carcinogenic effects from dietary administration of Brilliant Blue FCF. Reproductive and developmental toxicity studies did not show any effects on fertility or fetal toxicity. There is no evidence of this chemical being

cons idered a neurotoxic or immunotoxic chemical. Studies also show that this chemical is non

mutagenic. Metabolism studies indicate that primarily all of the Brilliant Blue FCF that was administered via gavage or via the diet in rats and mice was excreted un changed in the feces.

B. Toxicity Profile

Acute Toxicity

In one acute oral toxicity study in rats, reported in JECFA, the LD50 was determined to be greater than 2000 mg/kg body weight. In a subcutaneous study in mice, the LD50 was determined to be

4600 mg/kg body weight. No other acute studies were found in the literature.

Short term study

In one short

- term study, groups of 5 young rats were given injections of the color subcutaneously twice daily for three days. The test substance was administered as an aqueous solution at a level of 250 mg/kg body weight. The rats were sacrificed on the 4 th day. No oestrogenic activity was detected and no abnormalities were found. (JECFA, 1969)

Subchronic Oral Toxicity

In one subchronic study, O

- Sulfobenzaldehyde, one of the components of Brilliant Blue FCF, was

fed to groups of three week old rats (10 males and 10 females per group). The dose levels were 0, 0.25, 0.5, 1.0 and 2.0% (equivalent to 0, 125, 250, 500 and 10 00 mg/kg bw/day) for a period of 13

weeks. The no

- effect level for the test substance was determined to be 2% (equivalent to 1000 mg/kg bw/day)

. (JECFA, 1969)

In a second subchronic study, mice (sex and strain unspecified) were fed a dose of 1200 mg

Bril liant Blue over a time period of 19 days. No adverse effects were reported. (JECFA, 1969)

Subchronic Toxicity (injection)10 Subcut aneous injections of 1 ml of 0.8% aqueous solution of Brilliant Blue FCF given twice weekly to rats produced histological ch anges suggestive of subsequent sarcoma formation unassociated with chemical carcinogenic potential. (JECFA, 1969)

Subchronic daily injections of 2 mg Brilliant Blue over 30 days in mice (sex and strain unspecified) were well tolerated. Some animals show ed swelling of the liver and spleen after receiving a total of 4 mg. No other adverse effects were reported. (JECFA, 1969)

A 5% aqueous solution of Brilliant blue was given to cats via p arenteral injection at a dose of 1.0 g on the first day to 0.1 g on days nine through 18 . Administration of the test substance in this manner did not produce methaemoglobinaemia or Heinz bodies. (JECFA, 1969)

Chronic Toxicity (Oral)

Five beagle dogs received 2% Brilliant Blue FCF and 3 received 1% ( equivalent to 2000 or 1000 mg/kg bw/day of the same test material for a time period of one year. One dog at the top dose

level died after 17 d ays of treatment. A second dog , treated at the lower dose level, died after a time period of 46 weeks. The seco nd dea th was determined to be due to intercurrent virus infections. No clinical signs, gross lesions or histological abnormalities observed were attributable to treatment, but no further details were provided in the study. The NOAEL was 2% in the diet (e quivalent of 500 mg/kg bw/day. (Hansen et al., 1966 as cite d in EFSA, 2010)

In a study of Brilliant Blue FCF in mice (strain unspecified), a group of 57 males and 43 females were given 1 mg of the test substance per day in the diet. Observations were made over 500

- 700 days. No evidence of carcinogenic action was found. (Waterma n and Lignac, 1958 as cited in JECFA)

Brilliant Blue FCF was fed at a concentration of 4% (equivalent to 2000 mg/kg bw/day) in the diet to five male and five female rats for a time period of 600 days. Gross staining of the glandular stomach and some gran ular deposits in the stomach were found, but on evidence of tumors was observed. (William and Ivy, 1953 as cited in JECFA)

85 rats were fed a diet containing 0.1% of the color for their entire life span. The daily intake was

10

- 15 mg. No evidence of tum ors was found. (Klinke, 1955 as cited in JECFA)

Group of 24 weanling Osborne

- Mendel rats (12 males and 12 females) were fed the color at levels of 0, 0.5%, 1.0%, 2.0% and 5.0% (equivalent to 250, 500, 1000 or 2500 mg/kg bw/day ) for a time period of 2 years. The animals were weighed weekly and mortalities and abnormalities were recorded during the study. Blood counts, including white blood cell count, hemoglobin, hematocrit and differential cell count, were performed on 10 ani mals at each dose level. The blood counts were taken at 3, 11, 17 and 22 month intervals. Survivors were autopsied and organ weights were recorded for heart, liver, spleen, kidney and testes. No outward effects were observed in the study. Gross and mic roscopic examination revealed incidental lesions, unrelated to treatment, with no effect on tumor production. The statistical evaluation revealed no changes in11 organ weights, mortality or growth and hematology was normal. No evidence of treatment

- related neoplastic lesions were observed in this study The no observed adverse effect level (NOAEL) for this study is 5% (equivalent to 2500 mg/kg bw/day) Brilliant Blue FCF in the diet, the highest dose

tested. (Hansen et al., 1964 as cited in JECFA) and (Hansen et al., 1964 as cited in EFSA)

Four groups of 15 male and 15 female Wistar rats were fed diets containing 0, 0.03%, 0.3% and

3.0% (equivalent to 15, 150 or 1500 mg/kg bw/day) of the color for 75 weeks. Food consumption and efficiency reports were taken weekly during the first year of the study. Body weights were recorded weekly for the duration of the study. Red blood cell counts, hemoglobin estimations and hematocrit readings were done at intervals during the entire test period. At the end of the tes t, surviving rats were sacrificed and gross examination was completed for all organ and tissue samples. No effects on growth, food consumption, and food efficiency were found . Mortality was

increased in the 3% group, however this was determined to be a r esult of respiratory infection unrelated to administration of the test substance in the diet. Hematology findings were normal. The NOAEL for this study is 1500 mg/kg bw/day, the highest dose tested. (Mannell et al., 1962 as cited in JECFA) and (Mannell et al., 1962 as cited in EFSA)

Brilliant Blue FCF was administered in the diet to 360 mice , CD

- 1, COBS (ICR derived) from Charles River Breeding Laboratories. The test animals were 4 weeks old at the start of the study. Sixty /sex/group were tested at di etary levels of 0.5, 1.5 or 5% (equivalent to 661, 2064 and 7354 mg/kg bw/day in males and 819, 2562 and 8966 mg/kg bw/day in females). The test material used in this study was certified by US FDA and was found to contain 90% pure coloring and 10%

subsidi ary colorings, volatile chlorides and sulphates and uncombined intermediates. Both male and female test groups received the test material for 104 weeks. Clinical observations were conducted twice daily with a minimum of 5 hours between observations. Indi vidual body weight and food consumption was monitored weekly for the first 14 weeks, biweekly for weeks 16

- 26, and monthly for the remainder of the study. Detailed physical examination for signs to toxicity and palpation for masses were conducted weekly. Hematology testing was conducted on 10 randomly selected animals from each test group after 3, 6, 12, 18 and 24 months of the study. Hemoglobin, hematocrit, erythrocyte and total and differential leukocyte counts were the parameters examined. Autopsies were conducted on all animals that died during the study, as well as on animals that were sacrificed in a moribund condition or as scheduled. Histology was conducted on all control groups and on the highest dose group. Organ weights were recorded for the brain, gonads, kidneys, liver, spleen and thyroid and relevant statistical tests were performed. Detection of late appearing tumors was also possible because enough animals were alive at the end of the study. Blue stai ning of the hair, exposed skin and feces was noted in all test groups.

Mean food consumption remained similar for control and treated mice throughout the duration o f the study. Males and females in the 1.5% and 5% groups showed statistically significan t decreases in body weight. Hematological values remained similar in test groups and controls throughout the study. Occasional statistically significant decreases in average hemoglobin, hematocrit and leukocytes values in treated animals were noted, howe ver they were not considered toxicologically relevant by the authors. Lifetime exposure of mice to Brilliant Blue FCF as part of the diet did not result in consistent biologically significant, compound related adverse effects on behavior, morbidity, mort ality, hematology, general physical observations or tumor incidence. The decrease12 in mean body weights of treated mice were thought to be a result of the non

- nutritive nature of

Brilliant Blue FCF

.

The incidence of neoplasms in mice receiving Brilliant B lue FCF in the diet was evaluated throughout their lifetimes. Grossly detected masses and lesions from mice in the 0.5% and 1.5% dose groups were examined microscopically. Similar incidences of microscopic lesions were

observed in control and treated mic e. An increased incidence of hemangionmas in the spleen was found in the female 5% test group; however this was only significant with the Fisher test with unadjusted trend. The authors determined that the hemangiomas were considered spontaneous and

unr elated to exposure the test material . The no observed adverse effect level (NOAEL) for this study was determined to be 7354 mg/kg bw/day for male and 8966 mg/kg bw/day for female, the highest doses tested. (EFSA, 2010)

Charles River CD rats were treated with B rilliant Blue FCF in a chronic toxicity study coupled with a reproductive study. An in utero phase was conducted in which the test compound was administered to the F0 generation . For the chronic phase, 700 rats were treated at the start of the

stu dy (70/sex/group, including 2 control groups). Following completion of the in utero phase, a maximum of 2 rats per sex from each litter were randomly selected for the chronic phase. The offspring were fed at dietary levels of 0.1, 1 or 2% Brilliant Blue FCF (equivalent to 50, 514 and 1073 mg/kg bw/day for males and 62, 631 and 1318 mg/kg bw/day for females). The chronic phase required exposure for 30 months or to the po int where survival decreased to 12rats/sex in

any group

.

Observations of mortality, m orbidity and gross signs of toxicity were conducted twice daily with a minimum of 5 hours between observations. Body weights and food consumption for the F1 generation were measured weekly for the first 14 weeks, biweekly for the next 12 weeks, and

every 4 weeks for the remainder of the study. Weekly physical examinations were conducted to determine signs of toxicity and palpation for masses. 10 animals from each test group were randomly selected for hematology, clinical chemistry and urine analysis tests at 3, 6, 12, 18 and 24 months of the chronic phase and at the end of the study.

Examination of hematological parameters included hemoglobin, hematocrit, erythrocyte and total and differential leukocyte counts. Clinical chemistry tests included asp artate aminotransferase, alanine aminotransferase, alkaline phosphatase, blood urea nitrogen, fasting glucose, total protein and creatinine. Urine was examined for gross and microscopic appearance, specific gravity, pH, and for the presence of protein, g lucose, ketones, bilirubin and occult blood.

All animals that died spontaneously, were sacrificed in moribund condition or were sacrificed on schedule were subject to gross examination. Organ weights were measured in 10 rats/sex/group at

the interim kil ling and for all survivors at the end of the study. The organ weights measured were brain, gonads, kidneys, liver, spleen, and thyroid. Complete histology was conducted on all animals in the two control groups and for all animals in the highest dose grou p (2%). Ten rats/sex/group, randomly selected for an interim killing at 12 months were also subject to complete histological testing. Histological examinations included adrenal, aorta, liver, lung, lymph nodes, mammary gland, ovaries, pancreas, pituitary gland, nerve (sciatic), prostrate, salivary gland, seminal vesicles, skeletal muscles, skin, spinal cord, spleen, stomach, testes with epididymides,13 thymus, thyroid, trachea, urinary bladder, uterus and any tissue with gross change of an uncertain

nature. Middle and low dose group animals were examined grossly and histological examinations were conducted on any animals that revealed gross lesions or masses.

The treatment period was 116 and 111 weeks for male and female rats respectively. At the end of the study, 186 rats were sacrificed and 414 were sacrificed in extremis or died spontaneously or accidentally during the study.

Mean body weights for each treatment group were generally similar for control and treated males and females, with the exceptio n of the females in the 2% test group. After 90 weeks, females in the 2% group began a statistically significant downward trend. There was no dose related trend in

survival in either males or females; however survival was significantly decreased in 2% fe males compared to control groups from week 80.

All treatment groups showed blue staining of the hair, exposed skin and feces. There were no compound related differences between treated animals and control groups in terms of hematological clinical chem istry and urinanalysis parameters. Statistically significant differences in food consumption were noted males in the 2% group between weeks 27 and 66 and females

between weeks 79 and 90. Animals that were sacrificed at one year showed no compound related gross and histological changes after complete autopsies. Animals that died on test or were sacrificed at the end of the study also did not show compound related changes after complete autopsies. A variety of lesions, including neoplasm, were found during histopathological

evaluations in both treated and control animals. Similar incidences of lesions in control and treated rats were considered spontaneous. Lifetime exposure to Brilliant Blue FCF as part of the diet did not demonstrate carcinogenic effects. T he NOAEL is the dietary concentration of 2% (equivalent to 1072 mg/kg bw/day) for male rats and 1% (equivalent to 631 mg/kg bw/day) for females, based o n a 15% decrease in terminal mean body weight and decreased survival in high

dose females. (EFSA, 2010)

Reproductive Toxicity

Charles River CD rats were treated with Brilliant Blue FCF in a chronic toxicity study coupled with a reproductive study. Sixt y males and 60 females from the F0 generation were fed at dietary levels of 0.1, 1 or 2% Brilliant Blue FCF (equivalent to 50, 514 and 1073 mg/kg bw/day for males and 62, 631 and 1318 mg/kg bw/day for females) for 62 days at approximately 100 days of age. The F0 generation was housed, one rat/sex within the same treatment group for a period of 15 consecutive days to produce F1 litters. Body weights for females were recorded on gestation days 4, 14 and 21. There were no compound related effects on fertili ty, gestation, parturition, lactation, pup survival through weaning, or on number of live and stillborn pups. One female in each test group died, however, these deaths were not considered treatment related

. T he NOAEL is the dietary concentration of 2% (e quivalent to 107 3 mg/kg bw/day) for male rats and 2% (equivalent

to 1318 mg/kg bw/day) for females

. (EFSA, 2010)

Brilliant Blue FCF was fed to three successive generations of male and female rats at dose levels of 0, 10, 100, 300 or 1000 mg/kg bw/day. The first generation of parents were mated twice, the14 second generation parents were mated 3 times and the third generation of parents were mated twice. Dams delivered all offspring and raised them to weaning for all matings except the third mating of the F1b generation, when dams were sacrificed on day 19 of gestation and the uterine contents were examined. Offspring from various matings were selected to be parents of the next generation or were sacrificed and autopsied at weaning. Parental body weights and food consumption, as well as offspring survival and growth were recorded. Five rats/sex/dietary level for the parents of the second generation (F1b) and the offspring of the last generation (F3a) were autopsied, certain tissues were preserved, and se lected tissues from control animals and high dose group animals were examined histopathologically. The mean body weights of the high dose group were less than the control groups than for nursing offspring and in F1 and F2 males and females.

There were no treatment related effects on adult mortality, mating, pregnancy and fertility rates, lengths of gestation period, offspring survival or sex, litter survival or necropsy findings in animals sacrificed on day 19. There were also not macroscopic or microsco pic tissue alterations for F1b and F3a generations that were attributable to dietary administration of Brilliant Blue FCF The NOAEL for this study was 1000 mg/kg bw/day, the highest dose tested.

(EFSA, 2010)

Developmental Toxicity

Brilliant Blue FCF w as fed to groups of mated Long Evans rats fro m day 6 to 15 of gestation at 0, 200, 600, and 2000 mg/kg bw/day. Animals that survived the treatment period were sacrificed on day 20 and the number of corpor a lutea and uterine contents were recorded. Fetuses were

examined for malformations and two thirds of all fetuses were selected for v isualization of skeletal ossification, variations and anomalies. Administration of the color via the diet did not result in any fetal toxicity or anomalies. No furth er details were provided; therefore no NOAEL can be derived from this study. (EFSA, 2010)

In another developmental study, Brilliant Blue FCF was administered orally to mated female New Zealand White rabbits from day 6 to 18 of gestation at 0, 20, 60 or 2 00 mg/kg bw/day,

administered in v o lumes of 1 ml/kg for all treatment groups. Animals that survived the treatment period were sacrificed on day 20 and the number of corpora lutea and uterine contents were recorded. All fetuses were weighed, examined gros sly for externally visible defects, and carefully

dissected and examined for visceral anomalies. Fetuses were also cleared and stained with alizarin

red for visualization of skeletal ossification and variation. Administration of the color via the diet

di d not result in any fetal toxicity or anomalies. No further details were provided; therefore no NOAEL can be derived from this study. (EFSA, 2010)

Carcinogenicity (Oral)

Groups of 48 male and 50 female ASH/CS1 mice were given diets containing 0, 0.015, 0.15 or 1.5% (equivalent to 0, 20, 200, and 2000 mg/kg bw/day) of Brilliant Blue FCF for up to 80 weeks. Female animals in the highest test group showed evidence of signifi cantly reduced body weights, but the experiment did not establish whether this was toxic effect or as a result of unpalatability of the diet resulting in decreased food intake. Hematological examinations were conducted on both15 10 male and female mice at w eeks 13, 26 and 52 and on all surviving mice at week 80 showed no

consistent treatment related effects.

At week 13, mice in both sexes given 1.5% of the test substance showed significantly lower reticulocyte counts than those of the control animals. Ma les in the 0.15% group indicated a similar difference. The lowest dose group was not examined at this time point.

The relative stomach weights of males receiving 1.5% of the test substance were significantly reduced. Females receiving the highest test dose showed significantly increased brain weight, but this was viewed as a reflection of the reduced body weight, as the absolute weight of the organ was not affected by the treatment.

Full histopathological examination was performed on several tissue samples from mice in the control group and those given the diet containing 1.5% of the color (2000 mg/kg bw/day). Histopathological evaluation of tissue samples from the lower dietary level groups was limited to the liver, kidney and tissues seen to be ab normal at autopsy. The male test groups showed a significantly higher incidence of glomerulonephritis when compared individually with the control groups, but no consistent dose response was found. This effect was not found in the female test groups. Male mice at the highest dietary level showed an increase of mild liver changes, including incidence of foam cells and fatty change. Eight out of 44 males in the highest test group had tumors of the reticulolymphatic system (lymphosarcoma and reticulum cell ne oplasm), but this was not a statistically significant increase over the control incidence of 4 out of 44.

Tumor pathology seen in treated mice included squamous cell carcinoma of the stomach with 1 of 39 females at the highest dose tested and 0 of 44 in the controls. A single occurrence of thyroid

adenoma was found in the 30 mid

- dose males. Adrenal tumors, specifically one medeullary tumor in each of the mid and top dose males and one cortical nodule in the low dose males, were also found. A single occ urrence of mesenteric lipoma was found in low dose males and squamous cell

carcinoma of the skin was found in 1 of 26 mid

- dose females and 1 of 39 high dose females. None of the tumor incidences in the treated animals were individually statistically signi ficantly higher

than the control.

It was also reported in this study, that stomach and skin carcinoma occur spontaneously in mice, but no specific evidence was provided for the tested strain, ASH/CS1. Spontaneous occurrence of thyroid tumors was also r eported in mice. Kidney tumors (6 adenomas and 1 adenocarcinoma) were found in 7 of the 30 males in the 0.15% test group, while only a single tumor of this kind was found in the control group. Only 1 of 44 of the high

- dose males developed a kidney tumor, with none occurring in the other male test groups and all of the female test groups. It was also noted that kidney tumors are unusual in most strains of mice, but are consistently found in the ASH/CS1 strain. This study did not show any carcinogenic eff ects of Brilliant Blue FCF. (EFSA, 2010)

Mutagenicity

Five genotoxicity studies have been summarized in the European Food Safety Authority (EFSA) Panel on Food Additives and Nutrient Sources added to Food (ANS) document entitled, “Scientific16 Opinion of the Re

- evaluation of Brilliant Blue FCF (E 133) as a Food Additive, ” published in

2010. Based on the results of four of these studies, the panel concluded that Brilliant Blue FCF is not of concern with respect to genotoxicity. The results of the fifth st udy were determined to be

inconclusive.

Brilliant Blue FCF showed no evidence of mutagenicity when tested at concentrations of 0 to 10000 μg/plate in a number of standard Ames tests in Salmonella typhimurium strains TA98, TA100, TA1535, TA1537, TA1538 wit h or without metabolic activation.

In another test, Brilliant Blue FCF did not show any ability to induce mutation with tested in Escherichia coli at 10 mg/ml and liver S9 fraction.

In an in vivo mouse (strain unspecified) micronucleus assay, Brilliant Blue FCF was administered by one to four intraperitoneal injections at doses of 0, 500, 1000, or 2000 mg/kg body weight. No

micronucleus inductions w ere observed in bone marrow at any of the tested doses.

In another study Brilliant Blue FCF was tested a s 1 of 39 chemicals currently used as food additives. Groups of 4 ddY male mice were treated once, orally at doses up to the limit dose of 2000 mg/kg body weight. Comet assays were performed on glandular stomach, colon, liver, kidney, urinary bladder, lu ng, brain, and bone marrow. Assays were performed 3 and 24 hours after treatment. Brilliant Blue FCF did not yield a statistically significant increase in DNA damage in any organs.

Brilliant Blue FCF was tested in an Ames Salmonella/microsome test and in a chromosomal aberration test (Chinese hamster fibroblast cell line, CHL) as part of the testing of a group of 190 synthetic food additives. The test gave positive results for Brilliant Blue FCF, however, the results were accompanied by excessive vari ations in the osmolality of the culture medium. Therefore, the

results were not considered relevant.

Neurotoxicity

A study focused on synergistic interactions between commonly used food additives in an in vitro developmental neurotoxicity test using neuroblastoma cells. Developmental neurotoxicity was measured as an inhibition of neurite outgrowth in 24 hours. It was determined that Billiant Blue FCF (0.05

- 500 nM) in combination with L

- glutamic acid (0.5

- 100 μM) synergistically reduced neurite outgrowth length. (EFSA, 2010)

In addition to the study referenced above, neurotoxicity was not observed in multiple, long

- t erm toxicity studies.

Immunotoxicity

Immunotoxicity studies were not available for review. However, signs of Immunotoxicity were not observed in any of the studies that were submitted. EPA concluded that Immunotoxicity studies are not required at this time.17 Metabolism and Pharmacokinetics

In one study to determine the metabolism potential of Brilliant Blue FCF, 14 C Brilliant Blue FCF

radiolabelled on 7

- C methylene position (sodium salt) was given by gavage at a dose of 1.5 mg/kg/bw to female Sprague

- Dawley rats. About 0.27% of the test substance was absorbed through the gastrointestinal tract. The major route of excretion was through the feces with total recoveries being 92%. Intestinal absorption was estimated by urinary 14 C exc retion, expired 14 CO 2 and residual radioactivity in internal organs and tissues 96 hours after oral administration. Intestinal absorption in bile duct ligated rats was estimated in the same manner, resulting in 2.05% absorption. Mean fecal excretion was 97.28% and total recoveries averaged 99.38%. About 1.32% of the dose was eliminated via bilary excretion in bile

- cannulated animals within 96 hours.

14 CO2 excreted in expired air was 0.04% of the dose in intact animals and 0.01% of the dose in

bile

- cannu lated animals. The 24

- hour urine and bile samples evaluated using thin layer chromatography showed that approximately 95% of the excreted radioactivity was unaltered Brilliant Blue FCF with about 5% being an unidentified metabolite or degradation product. A minor peak of radioactivity with similar chromatographic characteristics was also detected in the dose solution standard at levels from 2 to 4% (over a 4

- month storage period) and may represent a decomposition product. ( Brown et al., 1980 as cited in E FSA, 2010)

In a second metabolism study, male and female rats (strain unspecified) were dosed by gavage with either 30 mcg/kg bw or 3 mg/kg bw of 14 C Brilliant Blue FCF . 99.9% of the lower dose was excreted in the feces within 72 hours, while 95.4% of th e high dose was excreted in the feces in the same time period. Less than 0.5% of the radioactivity was detected in the urine and no radioactivity was detected in the expired air. (Philips et al., 1980 as cited in EFSA, 2010)

Pregnant rats (strain unspeci fied) were given 3 mg Brilliant Blue FCF on day 8 of pregnancy. Pre

treatment with 30 mg/kg/day given in the diet for 21 days prior to gavage dosing with the labeled

material had no effect on the route of excretion or time taken to eliminate all of the ra dioactivity. Male mice and guinea pigs (strain unspecified) were given a single dose of 30 mcg/kg bw or 3 mg/kg bw Brilliant Blue FCF. It was determined that practically all of the administered dose was excreted in the feces and the urinary content of r adioactivity was less than 1% at both tested doses. Thin layer chromatography of the radioactive material extracted from the feces of rats and guinea pigs that had received a single dose of the color indicated that the test substance was not

metabolized d uring its passage through the gastrointestinal tract. The lack of gastrointestinal absorption and metabolism was confirmed by studies in guinea pigs and mice using isolated loops

of small intestines. (EFSA, 2010)

Metabolism data, as cited in JECFA, 1969 , indicated that Brilliant Blue FCF was administered orally to rats at 2% aqueous solution (200 mg per rat). In this test, almost the entire amount of the test substance was excreted unchanged in the feces within 40 hours after administration. In

another investigation, the presence of the color in the bile was observed in rats, rabbits and dogs after oral administration. In dogs, the amount did not exceed 5% of the administered dose. Administration of an aqueous solution of the color by stomach tube resulted in 89% excretion in the feces and no excretion in the urine. (JECFA, 1969)18 C. Toxicity Endpoint Selection

The available toxicity studies indicate that FD&C Blue No. 1 has a very low overall toxicity. The NOAEL is 1,072 mg/kg bw /day for male rats and 631 mg/kg bw/day for females, based on a 15% decrease in terminal mean body weight and decreased survival in high dose females in a chronic toxicity study coupled with a reproductive study. However, these results were not reproducib le in several other chronic longer duration studies at higher doses. Several long

- term studies indicate a higher NOAEL above the limit dose. Therefore, since no endpoint of concern was identified for the acute and chronic dietary exposure assessments and s hort and intermediate terms dermal and inhalation exposure assessments, a quantitative risk assessment for FD&C Blue No. 1 is not

necessary.

V. FQPA Safety Factor considerations

Section 408(b)(2)(c) of FFDCA provides that EPA shall apply an additional te nfold (10X) margin of safety for infants and children in the case of threshold effects to account for prenatal and postnatal toxicity and the completeness of the database on toxicity and exposure unless EPA

determines based on reliable data that a differen t margin of safety will be safe for infants and children. This additional margin of safety is commonly referred to as the FQPA safety factor (SF). In applying this provision, EPA either retains the default value of 10X, or uses a different

additional safet y factor when reliable data available to EPA support the choice of a different factor.

There is no evidence of qualitative or quantitative susceptibility of infants and children in the

available database.

As part of its qualitative assessment, the Agency did not use safety factors for assessing risk, and no additional safety factor is needed for assessing risk to infants and children. The available toxicity studies suggest low toxicity of FD&C Blue No. 1

. The toxicity database for FD&C Blue

No. 1 contains an acute oral toxicity study

, sub

- chronic and chronic toxicity studies, including carcinogenicity, reproductive and developmental toxicity studies. No reproductive or developmental toxicity was observed i n the modified reproduction study, 3

- generation reproduction study and developmental toxicity studies in rats and rabbits. The database also

contains mutagenicity studie s, neurotoxicity data and metabolism data. There is no indication

, based upon the avai lable data

, that FD&C Blue No. 1 is a neurotoxic or immunotoxic chemical or results in increased qualitative or quantitative susceptibility in infants or children. Based on this information, there is no concern, at this time, for increased sensitivity to i nfants and children to this chemical when used as inert ingredient in pesticides formulations.

VI . Exposure

A. Dietary Exposure19 Dietary exposure to FD&C Blue No. 1 can occur from eating food treated with pesticide formulations containing this inert ingredient. Dietary exposure can also occur from eating foods which contain FD&C Blue No. 1 as an ingredient. It is widely used as an ingredient in food products such as ice cream, bottled food coloring, icings, ice pops, dairy products, sweets and

drinks. However, since an endpoint of concern for risk assessment was not identified, a quantitative dietary exposure assessment for FD&C Blue No. 1 was not conducted

.

i. Acute exposure . No adverse effects attributable to a single exposure of FD&C Blue No. 1 was seen in the toxicity databases. Therefore, acute dietary risk assessments for FD&C Blue No. 1 is not required.

ii. Chronic exposure

. A chronic dietary exposure assessment was not conducted for FD&C Blue

No. 1 because no toxicological endpoint of concern was identified for chronic dietary exposure. The available studies/literature contained no evidence of a chronic dietary endpoint of concern.

iii. Dietary exposure from drinking water: Dietary exposure from drinking water to FD&C Blue

No. 1 can occur by drinking water that has been contaminated by run

- off from a pesticide treated

area

. Since an endpoint for risk assessment was not identified, a quantitative dietary exposure assessment from drinking water for FD&C Blue No. 1 was not conducted.

i v . Cancer exposure

. Based on the available toxicity data, FD&C Blue No. 1 is not expected to be carcinogenic. Therefore, a cancer dietary exposure ass essment is not necessary to assess cancer risk.

B

. Residential (Non

- Occupational) Exposure

From non

- dietary exposure

. The term “residential exposure” is used in this document to refer to

non

- occupational, non

- dietary exposure (e.g., for lawn and garden pest control, indoor pest control, termiticides, and flea and tick control on pets). The proposed use of FD&C Blue No. 1 as a seed

treatment /dye under 40 CFR 180.920 is not expected to result in residential exposure to this chemical. Residential exposure is possible based on other currently approved inert uses of this

chemical.

However, since there are no toxicological ef fects of concern occurring below 631 mg/kg bw/day, it is not necessary to conduct assessments of residential (non

- occupational) exposures and risks. There are no dermal or inhalation toxicological endpoints of concern to the Agency therefore quantitative assessments have not been conducted.

VII. AGGREGATE RISK

Acute Aggregate Risk

. There was no hazard attributable to a single exposure seen the in the toxicity database for the FD&C Blue No. 1 ; therefore an acute aggregate risk assessment is not required.

Short

- term Aggregate risk

. Short

- term aggregate exposure takes into account short

- term residential exposure plus chronic exposure to food and water (considered to be a background20 exposure level). FD&C Blue No. 1 could be used as an inert ingredient in pesticide products that

could result in short

- term residential exposure

.

Since there are no oral, dermal or inhalation toxicological endpoints of concern which occur below

the 631 mg/kg bw/day, it is not necessary to aggregate dermal and inhalation exposures with estimated dietary exposures.

Using the conservative exposure assumptions described above , EPA has concluded that the

combined sho rt

- term aggregated food, water exposures resul t in aggregate MOEs of > 100.

Residential Postapplication Exposure

Assessments of Residential Post

- application exposure are not necessary since there are no toxicological endpoints of concern at doses less than 631 mg/kg bw/day

.

VIII. A GGREGATE RISK ASSESSMENTS AND RISK CHARACTERIZATION

Acute Aggregate Risk

There was no hazard attributable to a single exposure seen the in the toxicity database for FD&C

Blue No. 1 therefore, an acute aggregate risk assessment is not required.

Short

- Term/Intermediat e

- Term Aggregate Risk

Short

- term and intermediate

- term aggregate risk assessments for FD&C Blue No. 1were not conducted. The assessments are not necessary since there are no dermal or inhalation toxicological

effects of concern below 631 mg/kg bw/day. There are no risks that exceed the Agency’s level of

concern.

IX. Occupational Exposure

Typically, seeds used in the propagation of agricultural crops are treated with one or more pesticide products. Seed treatment is conducted to help mitigate the de leterious effects of disease organisms or insects upon the seeds. By regulation, seeds treated with a pesticide must also be treated with a “colorant” to plainly indicate that the seeds are, in fact treated with a pesticide. A colorant is applied so that treated seeds may only be used for propagation (planting) and may NOT be used for food or feed processing.

Active (pesticide) ingredients used for seed treatment are evaluated by the EPA for possible adverse effects upon the environment or upon workers who may handle treated seeds. In like manner, the Agency also evaluates the “inert” ingredients which are concurrently applied to seeds. Such “inert” ingredients might be “carriers” or “stickers” or colorants which are applied along with any given activ e ingredient or ingredients.

In the case of FD&C Blue No. 1, the Agency has reviewed the available toxicological information. The data indicate that there are no adverse toxicological effects which occur at21 exposures less than 631 mg/kg bw/day, therefo re a quantitative risk assessment is not necessary

and is not conducted.

X. Cumulative Exposure

Cumulative effects from substances with a common mechanism of toxicity

. Section 408(b)(2)(D)(v) of FFDCA requires that, when considering whether to establish, modify, or revoke a tolerance, the Agency consider “available information” concerning the cumulative effects of a particular pesticide's residues and “other substances that have a common mechanism of toxicity.”

EPA has not found FD&C Blue No. 1 to share a common mechanism of toxicity with any other substances, and FD&C Blue No. 1does do not appear to produce a toxic metabolite produced by other substances. For the purpos es of this tolerance action, therefore, EPA has assumed that the

FD&C Blue No. 1 do es not have a common mechanism of toxicity with other substances. For information regarding EPA's efforts to determine which chemicals have a common mechanism of toxicity an d to evaluate the cumulative effects of such chemicals, see EPA's website at http://www.epa.gov/pesticides/cumulative

.

X I

. Enviromental Fate

In terms of terrestrial fate, Brilliant Blue is expected to be mobile in soil as it is expected to exist almost entirely in anion form in the environment

. Generally, anions do not adsorb more strongly to soils containing organic carbon and clay th an their neutral counterparts . Volatilization from

moist soil i s also not expected because anions do not volatilize. Brilliant Blue is not expected to volatilize from dry soil surfaces because it is a salt and salts do not volatilize

. B iodegradation in soils is not expected to be an important environmental fate proces s due to activiated sludge and chemical precipitation treatments of wastewater containing anionic dyes resulting in dye removal efficiencies of the than 25%. (Douchette, W.J. as cited in HSDB).

In terms of aquatic fate, Brilliant Blue is not expected to adsorb to suspended solids and sediment and will exist almost entirely i n anion form in the environment. Generally, anions do not adsorb more strongly to soils containing organic carbon and clay th an their neutral counterparts

. Volatilization from water s urfaces is also not expected . B ioconcentration in aquatic organisms is expected to be low, based on the ioni c character of the compound

. (Douchette, W.J. a s cited in HSDB)

In terms of atmospheric fate, model predictions of gas/particle partitioning of se mivolatile organi c compounds in the atmosphere , Brilliant Blue, a salt, indicate that it is expected to exist solely in the particulate phase in the ambient atmosphere be cause salts are nonvolatile

. Brilliant Blue may be susceptible to direct photolysis by sunlight

. ( Bidleman , T.F., as cited in HSDB)22 X I I

. Ecological Effects

TABLE 4 . Ecotoxicity of FD&C Blue No. 1 Organism Test

Material Duration Endpoint References (*)

Colinus virginianus ( Northern Bobwhite Quail) 8 days L C 50 > 5620 ppm

US EPA,

ECOTOX Database, 2013 Anas platyrhynchos ( Mallard Duck) 8 days LC 50 > 5620 ppm

Anas platyrhynchos ( Mallard Duck) 14 days LD 50 > 2250 ppm

Colinus virginianus ( Northern Bobwhite Quail) 14 days LD 50 > 2250 ppm

Daphnia magna ( Water Flea ) 2 days EC50> 97000 ug/L

Table 5 . Acute Toxicities to salmonids of Basacid Blue *

Dilution Water Fish Species LC50 (mg product/L)

24 hour 48 hour 72 hour 96 hour Soft (city) Oncorhynchus kisutch (Silver Salmon, Coho Salmon) 1154 332 332 332

Oncorhynchus tshawytscha (Chinook Salmon) 649 721 412 370

Oncorhynchus keta (Chum Salmon) 1130 801 452 427

Oncorhynchus gorbuscha (Pink Salmon) 680 680 420 420

Oncorhynchus mykiss (Rainbow Trout) 1030 972 818 412

Oncorhynchus nerka (Sockeye Salmon) 700 700 201 180

Intermediate (reconstituted) Oncorhynchus kisutch Silver Salmon, Coho Salmon 2038 1261 1144 1144

Oncorhynchus gorbuscha (Pink Salmon) 1126 1126 619 619

Oncorhynchus mykiss (Rainbow Trout) 2114 1421 1370 1370

Hard (lake) Oncorhynchus kisutch Silver Salmon, Coho Salmon 569 392 377 377

Oncorhynchus gorbuscha (Pink Salmon) 674 265 252 23923 Oncorhynchus mykiss ( Rainbow Trout ) 3457 1525 1474 1474

* Wan,M.T., R.G. Watts, and D.J. Moul, 1991 as cited in US EPA ECOTOX Database ( http://cfpub.epa.gov/ecotox/ )

The data suggest that FD&C Blue No. 1 has relatively of low toxicity aquatic organisms.

XI I I

. D etermination of Safety

.

Based on the lack of toxicological effects of concern and upon the conditions discussed above

, the EPA concludes that there is a reasonable certainty that no harm will result to the general population, or to infants and children from aggregate exposure to residues of the FD&C Blue No. 1 when used as an inert ingredient in pesticide formulations under 40 CFR 180. 920

.

X IV . Enviromental Justice

Potential areas of environmental justice concerns, to the extent possible, were considered in this human health risk assessment, in accordance with U.S. Executive Order 12898, "Federal Actions to Address Environmental Justice in Minority Populations and Lo w

- Income Populations," http://www.eh.doe.gov/oepa/guidance/justice/eo12898.pdf ).

As a part of every pesticide risk assessment, OPP considers a large variety of consumer subgroups according to well

- established procedures. In line with OPP policy, HED estimates risks to population subgroups from pesticide exposures that are based on pat terns of that subgroup’s food and water consumption, and activities in and around the home that involve pesticide use in a residential setting. Extensive data on food consumption patterns are compiled by the USDA under the Continuing Survey of Food Intake by Individuals (CSFII) and are used in pesticide risk assessments for all registered food uses of a pesticide. These data are analyzed and categorized by subgroups based on age, season of the year, ethnic group, and region of the country. Additionally, OPP is able to assess dietary exposure to smaller, specialized subgroups and exposure assessments are performed when conditions or circumstances warrant. Whenever appropriate, non

- dietary exposures based on home use of pesticide products and associated ri sks for adult applicators and for toddlers, youths, and adults entering or playing on treated areas postapplication are evaluated. Further considerations are currently in development as OPP has committed resources and expertise to the development of speci alized software and models that consider exposure to bystanders and farm workers as well as lifestyle and traditional dietary patterns among specific subgroups.

X V . Human Studies

This assessment does not rely on any studies in which human subjects w ere intentionally exposed to a pesticide.

XVI

. International limits24 In making its tolerance decisions, EPA seeks to harmonize U.S. tolerances with international

standards whenever possible, consistent with U.S. food safety standards and agricultural practices.

EPA considers the international maximum residue limits (MRLs) established by the Codex Alimentarius Commission (Codex), as required by FFDCA section 408(b)(4). The Codex Alimentarius is a joint Uni ted Nation Food and Agriculture Organization/Wor ld Health Organization food standards program, and it is recognized as an international food safety

standards

- setting organization in trade agreements to which the United States is a party. EPA may establish a tolerance that is different from a Codex MRL; however, FFDCA section 408(b)(4) requires that EPA ex plain the reasons for departing from the Codex level. The Codex has not

established a MRL for FD&C Blue No. 1.

References

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