2 FD&C Green No. 3 is currently approved for use as an inert ingredient in non-food use pesticide products. The Joint FAO/WHO Expert Commit tee on Food Additives (JECFA) has assigned maximum acceptable daily intakes (ADIs) of 25 mg /kg body weight for FD&C Green No. 3. It is currently used as a biological stain; colo r additive in food, drugs and cosmetics.

FD&C Green No. 3 is not acutely toxic via the oral route in rats and dogs given the median lethal doses (LD 50 ) were greater than 2000 mg/kg body wei ght and 200 mg/kg body we ight, respectively. Groups of 60 male and female Charles-River SC- mice, including 120 control animals, were fed diets containing 0, 0.5, 1.5 or 5.0% (equivalent to 0, 750, 2250 and 7500 mg/kg bw/day) Fast Green FCF beginning at 43 days of age for appr oximately 24 months. There were no treatment related effects on mortality in this study. The mean body weights for the females in the 5% dose group were consistently lower than the controls. Histological exam ination of all animals did not reveal any treatment related lesions. There wa s also no difference betw een control and treated animals in terms of the incidence of benign a nd malignant neoplasms. The no observed adverse effect level (NOAEL) was 5% in the diet (equivalent of 7500 mg/kg bw/day), the highest dose

tested.

Multiple long-term studies in mice, dogs and ra ts fed diets containing Fast Green FCF were conducted. Microscopic examination revealed no tr eatment related lesions attributable to feeding of the color in any of the studies. There were also no treatment relate d effects on growth or mortality.

A carcinogenicity study with a in utero phase wa s conducted with Charles-River albino rats. Groups of 60 male and female rats of the F 0 generation were fed diets containing 0, 1.25, 2.5 or 5.0% (equivalent to 0, 625m 1250 and 2500 mg/kg bw/d ay) Fast Green FCF for 2 months prior to mating and throughout gestation and lactation. Ther e were also 120 control animals used in this study. Following the reproductive phase, a maximum of 4 rats/sex/litter we re randomly selected

from the F 1 generation for the long-term carcinogenicity study. Seventy animals of each sex/group

were fed the test substance at the same concen trations as the parent generation. During the premating period, there were no treatment-related effects on mo rtality or body-weight gain. However, there was a dose-relate d increase in food consumption. There were no treatment-related effects on the number of successful pregnancies or pup viability at birth, but pup mortality was increased in the 5% dose group during day 4-14 of lactation. Mean pup weight was reduced in all test groups, most distinctly in the high-dose group. In the F 1 generation mortality was slightly higher in all treatment groups as compared to cont rol groups. However, it did not occur in a doesrelated manner. At interim sacrifice the mean and relative thyroid weights were elevated in highdose males and relative kidney weights were elevated in high-dose females. At termination, males and females showed elevated thyroi d weights. Kidney weights were elevated in both sexes of the 5% group. “Histopathological examination reveal ed an increased inci dence of urothelial hyperplasia in treated males and of urinary bladde r transitional cell/urothe lial neoplasms in males of the 5%-dose group.” It was c oncluded that the obser vation of only three neoplasms in the highdose male group, two of which were of questiona ble neoplastic character, and the absence of evidence of a pre-neoplastic process in the obser ved hyperplasia, establis hed that there is no indication of a neoplastic effect on the urinary bl adder from administration of Fast Green FCF.” The NOAEL for carcinogenicity was 5% in the diet (equivalent of 2500 mg/ kg bw/day; the highest dose tested). No reproductive toxicity was observed at doses up to 5% in the diet (equivalent to3 2500 mg/kg bw/day). The NOAEL for systemic toxicity is parental animals was 2.5% in the diet (equivalent to 1250 mg/kg bw/day) based on decreased in food consumption, increased in thyroid and kidney weights seen at the LOAEL of 5% in the diet. The NOAEL for off spring toxicity was 2.5% in the diet (equivalent to 1250 mg/kg bw/day) based on decreased in pup body weight and pup mortality seen at the L OAEL of 5% in the diet.

A 3-generation reproduction study was completed on Fast Green FCF in Long-Evans rats at dose levels of 0, 10, 100, 300 or 1000 mg/kg bw/day. No treatment related effects on food consumption, body weight, adult mortality, mating pe rformance, pregnancy and fertility rates, gestation length, offspring surviv al, weights and sex, litter survival , resorption rates, or necropsy findings. There were also no macroscopic or mi croscopic tissue abnormalities attributable to

treatment. The NOAEL wa s 1000 mg/kg bw/day.

In a Salmonella/microsome assay, Fast Green FCF was determined to be non-mutagenic. Negative

results were also found in b acterial DNA repair tests.

Immunotoxicity and neurotoxicity studies were not available for review. However, signs of immunotoxicity and neurotoxicity were not observed in any of the studies that were submitted and

EPA concluded that studies are not required at this time.

The metabolism potential of Fast Green FCF was test ed in rats and dogs. Almost all of the color was excreted unchanged in the feces of the rats a nd no color was found in the urine. The color did not exceed 5% of the given dose in the bile of the dogs. After feeding, th e color was found in the bile of rats, but not in the urine. It was concluded that th e amount found in the bile provides an

estimate of the amount absorbed thro ugh the gastrointestinal tract.

The available toxicity studies indicate that FD&C Green No. 3 has a very lo w overall toxicity. The lowest NOAEL in the database is 1000 mg/kg bw /day. In the carcinogenicity study with an in utero phase, the effects on the pups (decrease d body weights and pup mortality) and kidney and thyroid toxicity in adults were observed at 5% in diet (equivale nt to 2500 mg/kg/day). Since these

effects were observed at 2.5 times the limit dose of 1000 mg/kg/day, there are low concerns for the hazard. Since no endpoint of concern was identif ied for the acute and chronic dietary exposure

assessments and short- and intermediate-term de rmal and inhalation exposure assessments, a quantitative risk assessment for FD &C Green No. 3 is not necessary.

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 toxici ty FD&C Green No. 3. The toxicity database for FD&C Green No. 3 contains acute oral toxicity studies and chronic t oxicity studies, including carcinogenicity and reproductive toxi city studies. The database also contains mutagenicity studies and metabolism data. There is no indication, base d upon the available data, that FD&C Green No.

3 is a neurotoxic or immunotoxic chemical or re sults in increased qual itative or quantitative susceptibility in infants or childre n. 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.4 Dietary exposure to FD&C green No. 3 can occur from eating food that has come in contact with surfaces treated with pesticide fo rmulations containing this inert ingredient. Dietary exposure to FD&C Green No. 3 can also occur from eating f oods which contain FD&C Green NO. 3 as an ingredient. Non-dietary exposur e from the proposed use is not expected, however non-dietary exposure may occur from other approved uses of this chemical. Occupational exposure is also possible. However, since there are no toxicologi cal effects of concern identified in the available database, it is not necessary to conduct assessmen ts of residential (non-occupational) exposures

and risks. There are no dermal or inhalation to xicological endpoints of concern to the Agency; therefore quantitative assessmen ts have not been conducted.

EPA has not found FD&C Green No. 3 to share a common mechanism of toxicity with any other substances, and FD&C Green No. 3 does do not a ppear to produce a toxic metabolite produced by other substances. For the purposes of this tolera nce action, therefore, EP A has assumed that the

FD&C Green No. 3 does not have a common mech anism of toxicity with other substances.

In terms of terrestrial, aquatic a nd atmospheric fate, FD&C Green No. 3 is expected to be mobile in soil as it is expected to exist almost entirely in anion form in the environment. Anions do not adsorb more strongly to soils co ntaining organic carbon a nd clay than their neutral counterparts.

Volatilization from moist soil and wa ter surfaces is not expected. Due to the ionic nature of this chemical, bioconcentration in aquatic organisms is not expected. Ecological modeling indicates

that FD&C Green No. 3 ha s potentially low toxicity to aquatic organisms.

Potential areas of environmental ju stice concerns, to the extent possi ble, were considered in this human health risk assessment, in accordance w ith U.S. Executive Order 12898, "Federal Actions

to Address Environmental Just ice in Minority Populations an d Low-Income Populations," http://www.eh.doe.gov/oepa/ guidance/justice/eo12898.pdf

.

Based on the lack of toxicological effects of c oncern 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 Green No. 3 when used as an inert ingredient in pesticide formulations under 40 CFR 180.940(a)

.

I . BACKGROUND

This is the decision document for FD&C Green No. 3 (CAS Reg. No. 2353-45-9). The firm Exponent (1150 Connecticut Ave., NW; Suite 1100; Washington, DC 20036), on behalf of Ecolab (370 N. Wabasha St., St. Paul, MN 55102) has submitted a request to the Environmental Protection Agency (EPA or “the Agency”) to ex empt from the requirements of a tolerance FD&C Green No. 3 when used as an inert ingredient in antimicrobial formulations, for use on food contact surfaces in public eating places, dair y processing equipment, and food processing equipment and utensils under 40 CFR 180.940(a). The chemical is also refe rred to as Fast Green FCF in this document, as this name is synonymous with FD&C Green No. 3. FD&C Green No. 3 was evaluated by the Joint FAO/WHO Expert Committee on Food Additives (JECFA) multiple times, most recently in 1987. JECFA established an acceptable daily intake (ADI) of 25 mg/kg body weight.5 II. Inert Ingredient Profile

A. Summary of Uses

1. Proposed Use Pattern

The proposed use pattern requested in this pe tition for FD&C Green No. 3 is as an inert ingredient in antimicrobial formulations, fo r use on food contact surfaces in public eating places, dairy processing equipment, and food processing equipment and utensils under 40 CFR 180.940(a), without limitation.

2. Inert uses

FD&C Green No. 3 is currently approved for use as an inert ingredient in non-food use pesticide products.

3. Other uses

The Joint FAO/WHO Expert Committee on Fo od Additives (JECFA) have assigned a maximum acceptable daily intake (ADIs) for FD&C Green No. 3. It is currently used as a biological stain; color additive in food, drugs and cosmetics.

Table 1. FDA Approved Color Additive Uses

Chemical 21 CFR 74 Uses

FD&C Green No.

3 74.203 Coloring in food, including dietary supplements 74.1203 Coloring in drugs 74.2203 Coloring in cosmetics6 III. Physical Chemical Properties

Some of the physical and chemical characteristic s of FD&C Green No. 3, along with its structure and nomenclature, are found in Table 3.

Table 2. Physical and Chemical Properties of FD&C Green No. 3

Parameter Value Source

Structure ChemIDplus Lite, 2013

CAS Chemical Name Benzenemethanaminium, N-ethylN-(4-((4-(ethyl((3- sulfophenyl)methyl)amino)phenyl)( 4-hyd- roxy-2- sulfophenyl)methylene)-2,5-cyclohexadien-1-ylidene)-3-sulfo-, hydroxide, inner salt, disodium

salt ChemIDplus Lite, 2013

Chemical Formula C37-H36-N2-O10-S3 ChemIDplus Lite, 2013

Synonyms C.I. Food Green 3, disodium salt,

Fast Green FCF ChemIDplus Lite, 2013

CAS # 2353-45-9 ChemIDplus Lite, 2013

SMILES Notation C(=C1\C=C\C(=[N+](\Cc2cc(ccc2) S(=O)(=O)[O- ])CC)C=C1)(/c1c(cc(cc1)O)S(=O)( =O)O)c1ccc(cc1)N (Cc1cc(ccc1)S(= O)(=O)O)CC.[Na].[Na] ChemIDplus Lite, 2013

Color/Form Dark green powder or granules with

a metallic luster. Red to brownviolet powder for dark green crystals PubChem, 2013

Molecular Weight 808.12 PubChem, 2013

Water Solubility 20.0 g/100 mL at 25 deg C (2.0X10+5 mg/L) PubChem, 2013

Melting Point (°C) 290 °C PubChem, 2013 Atmospheric OH Rate 4.18E-10 cm3/molecu le-sec (est) ChemIDplus Lite, 20137 Constant

Partition Coefficient (usually the Log P or Kow) Log P= -5.420 (est) Log Kow= -3.22 (est) ChemIDplus Lite, 2013

IV. Hazard Assessment

A. Summary of Toxicity Data.

FD&C Green No. 3 is not acutely toxic. Long term studies of the effects of the color administered in the diet to rats, mice and dogs did not indi cate any significant toxi c effects. A long term carciongencity study did not show any carcinogenic effects from diet ary administration of FD&C Green No. 3. A reproductive study did not show any effects on fer tility or fetal abnormalities. There is no evidence of this chemical being co nsidered a neurotoxic or immunotoxic chemical. Studies also show that this chemical is non-mutage nic. Metabolism studies indicate that primarily

all of the FD&C Green No. 3 that was administer ed via the diet in rats and mice was excreted unchanged in the feces.

B. Toxicity Profile

Acute Toxicity

In one acute oral toxi city study in rats, reported in JECFA, the LD 50 was determined to be greater than 2000 mg/kg body weight. In a second study with dogs the LD 50 was greater than 200 mg/kg.

No other acute studies were found in the literature.

Subchronic Toxicity

No subchronic toxicity studies are available in the database.

Chronic Toxicity (Oral)

Groups of 60 male and female Charles-River SC- mice, including 120 control animals, were fed diets containing 0, 0.5, 1.5 or 5.0% (equivalent to 0, 750, 2250 and 7500 mg/kg bw/day) Fast Green FCF beginning at 43 days of age for approximately 24 months. Hematological examinations were conducted at 3, 6, 12 and 18 months on 10 animals/sex/group. Post-mortem

examination was conducted on all animals that di ed during treatment or were killed in moribund

condition. Animals that survived to termination were also subject to post-mortem examination. All survivors from the control and 5% dose groups were subject to hist ological examination of tissues including adrenals, aorta, bone and marrow (femur), brain (3 sections), eyes (with optic nerve), gall bladder, gastrointe stinal tract (esophagus, stomac h, duodenum, ileum, caecum, colon), heart, kidneys, liver, lung, lymph nodes (mesen teric and mediastinal), mammary gland, nerve (sciatic), ovaries, pancreas, pitu itary, prostate, salivary gland, se minal vesicles, skeletal muscle,8 skin, spinal cord, spleen testes with epididym ides, thymus, thyroid/para thyroid, trachea, urinary bladder, uterus, and gross lesions and tissue masses. All animals that that died or were killed in extremis from these groups were subject to the same histological ex aminations. All animals in the lower dose test groups were examined for signs of gross changes and tissue masses.

There were no treatment related effects on morta lity in this study. The mean body weights for the females in the 5% dose group were consistently lower than the controls. Mean body weights for males were consistently lower than the controls at weeks 52 and 78. There were no other effects on body weight noted in the study. In high-dose males at 18 months, there were slight reductions in hemoglobin, hematocrit and erythrocyte counts. However, there were no other consistent or dose-related changes observed. Histological exam ination of all animals did not reveal any

treatment related lesions. There was also no di fference between control and treated animals in terms of the incidence of benign and maligna nt neoplasms. The NOAEL was 5% in the diet (equivalent of 7500 mg/kg bw/day), the highest dose tested. (JECFA, 1986)

In a second study with 50 male and 50 female C3HeB/FeJ mice were fed diets containing 1.0 or 2.0% Fast Green FCF for 2 years (equivalent to 1500 and 3000 mg/kg bw/day). One hundred mice

of each sex served as controls. After 78 weeks, 56 controls, 27 an imals in the 1.0% test group and 17 animals in the 2.0% test group still survived. Microscopic examination reveled no treatment

related lesions attributable to feed ing of the color. (JECFA, 1986)

In a third study in dogs, 4 beagle dogs/ group, equally divided by se x, were fed Fast Green FCF for a time period of 2 years at 0, 1.0 and 2.0% in the diet (equivalent to 250 and 500 mg/kg bw/day). Histopathology attributable to the color was limited to green blobs of pigmen t in the renal cortical tubular epithelial cytoplasm of a male dog at th e high-dose level. A fe male dog at the high-dose level showed slight interstitia l nephritis and slight bone marrow hyperplasia. (JECFA, 1986)

In a fourth study in rats, groups of 50 weanling Osborne-Mendel rats, even ly divided by sex, were fed diets containing 0, 0.5, 1.0, 2.0 or 50.% Fast Green FCF for 2 years (equivalent to 250, 500, 1000 or 2500 mg/kg bw/day). There were no treatm ent related effects on growth or mortality. Microscopic examination revealed no lesions attributed to the f eeding of the test substance. (JECFA, 1986)

In a fifth study in rats, the co lor was fed at a dietary level of 4.0% (equivalent to 2000 mg/kg bw/day) for a time period of 18 to 20 months. This resulted in gross stai ning of the forestomach, glandular stomach, small intestine and colon. Gra nular deposits were also noted in the stomach. No tumors were observed. (JECFA, 1986)

Carcinogenicity (Oral)

A carcinogenicity study with a in utero phase wa s conducted with Charles-River albino rats. Groups of 60 male and female rats of the F 0 generation were fed diets containing q, 1.25, 2.5 or 5.0% (equivalent to 0, 625m 1250 and 2500 mg/kg bw/d ay) Fast Green FCF for 2 months prior to mating and throughout gestation and lactation. Ther e were also 120 control animals used in this study. Following the reproductive phase, a maximum of 4 rats/sex/litter we re randomly selected9 from the F 1 generation for the long-term carcinogenicity study. Seventy animals of each sex/group

were fed the test substance at the same concentr ations as the parent generation. After 12 months, an interim kill was carried out for 10 rats of each sex per group. The remaining animals continued to receive the test diet for 29 months (males) or 31 months (females). Hematology, clinical chemistry tests and urinalysis were performed on 10 rats/sex/group at 3, 6, 12, 18 and 24 months. Gross autopsies were performed on all animals. Histological examination of tissues was

conducted on all animals killed at interim sacrific e and all survivors from the control and 5% dose groups, as well as all animals that died or were killed in extremis for these dose groups. Histological examination of tissues included adre nals, aorta, bone and marrow (femur), brain (3 sections), eyes (with optic nerve), gall bladde r, gastrointestinal tr act (esophagus, stomach, duodenum, ileum, caecum, colon), heart, kidneys, liver, lung, lymph nodes (mesenteric and mediastinal), mammary gland, nerve (sciatic), ovaries, pancreas, p ituitary, prostate, salivary gland, seminal vesicles, skeletal muscle, skin, spinal cord, spleen, testes with epididymides, thymus, thyroid/parathyroid, trachea, urin ary bladder, uterus, and gross lesions and tissue masses. All animals in the lower dose test groups were examin ed for signs of gross changes and tissue masses.

During the premating period, there were no treatm ent-related effects on mo rtality or body-weight gain. However, there was a dose-related increase in food consumption. There were no treatmentrelated effects on the number of successful pregna ncies or pup viability at birth, but pup mortality was increased in the 5% dose group during day 4-14 of lactation. M ean pup weight was reduced in all test groups, most distin ctly in the high-dose group.

In the F 1 generation mortality was sligh tly higher in all treatment groups as compared to control groups. However, it did not occur in a does-related manner. In spite of el evated food intake in high-dose males, mean body weights were consiste ntly lower than controls. Fasting blood glucose levels were elevated in females in all groups at 3 and 12 months. Females in the 1.25 and 2.5% groups showed elevated levels at 18 months. Males in all treatment groups showed elevated

levels at 12 and 18 months.

At interim sacrifice (12 months), the mean and re lative thyroid weights we re elevated in high-dose males and relative kidney weights were elevated in high-dose females. At termination, males in the 2.5 and 5% dose groups and females of the 5% group showed elevated thyroid weights. Kidney weights were elevated in both sexes of the 5% group. Females in the 2.5% group also showed increased kidney weights. There were no treatment related effects seen in urinalyses, hematology determinations, physical observations or opthanlmology.

“Histopathological examination reve aled an increased incidence of urothelial hyperplasia in treated males and of urinary bladder transitional cell/uroth elial neoplasms in males of the 5%-dose group.

The overall incidences in males are summarized below:”

Group Control 1 Control 2 1.25% 2.5% 5%

Number

examined 58 61 58 55 60

Number with neoplasia 1 2 1 2 5

Number with hyperplasia 1 4 7 10 310 “Non-statistically-signifi cant increases in testicular Leydig cell tumors and neoplastic nodules in

the liver were also observed. When time-to-tumor analysis was performed on pathology incidence data, the increased incidence of bladder tumors was confirmed and the inci dence of several other tumor types showed statistically-significant differe nces related to treatme nt, including neoplastic nodules in the liver (males and females), fema le mammary adenomas and pituitary adenomas, male parathyroid adenomas, male thyroid medullary carcinomas, female uterine leiomyosarcomas, and male testicular interstitial/Leydig cell tumors.”

“Of the non-neoplastic pathology, ch ronic nephropathy was a common finding in all groups but the severity was greater in females in the 5%-dose gro up. Other lesions did not a ppear to be related to treatment.” (JECFA, 1986)

The results of the carcinogenicity study were re-evaluated in 1987. “A review of the histopathological data from the ra t oral carcinogenicity study wa s performed from which it was concluded that inappropriate statis tical tests had been performed on some of the data by the testing laboratory. Using appropriate stat istical procedures the reviewers concluded that there were no significant differences in tumour incidence be tween combined control groups and the high-dose group with respect to tumours of the liver, test es, or thyroid; the ti ssues from the low- and intermediate-dose groups were not examined. In a b lind review of the slides of the bladders from all dose groups, only three prol iferative lesions, two benign (p apilloma) and one malignant (carcinoma), were observed in bladders of male ra ts in the high-dose group and a treatment-related

increase in the incidence or se verity of transitional cell hyperp lasia was not detected. It was concluded that the observation of only three ne oplasms in the high-dose male group, two of which were of questionable neoplastic character, and the absence of ev idence of a pre-neoplastic process in the observed hyperplasia, estab lished that there is no indicati on of a neoplastic effect on the urinary bladder from administration of Fast Green FCF.” The NOAEL for carcinogenicity was 5% in the diet (equivalent of 2500 mg/kg bw/day; th e highest dose tested). No reproductive toxicity was observed at doses up to 5% in the diet (equivalent to 2500 mg/kg bw/day). The NOAEL for systemic toxicity is parental animals was 2.5% in the diet (equivalen t to 1250 mg/kg bw/day) based on decreased in food consumption, increase d in thyroid and kidney weights seen at the LOAEL of 5% in the diet. The NOAEL for off spring toxicity was 2.5% in the diet (equivalent to 1250 mg/kg bw/day) based on decreased in pup body weight and pup mortality seen at the LOAEL of 5% in the diet. (JECFA, 1987)

Reproductive Toxicity

A 3-generation reproduction study was completed on Fast Green FCF in Long-Evans rats at dose levels of 0, 10, 100, 300 or 1000 mg/kg bw/day. Th e first generation parents (10 males and 10 females) were given the appropria te dose of the test substance 2 weeks prior to mating with dosing continuing throughout gestation, lactation, and post -weaning phases for 3 successive generations.

The F 0 generation rats were mated twice. The F 1a litters were necropsied at weaning and 10

males and 10 females from the F 1b litters were used for breeding.11 After and 80-day growth period, animals from the F 1b generation were mated 3 times. And the offspring from the F2a and F2b generations were treated in the same manner as the F1a and F1b animals. After completion of the third mating, half of the pregnant dams were sacrificed on day 19 of gestation with the uterine contents being examined for to tal embryos/resorption sites and corpora lutea per ovary. The othe r half were allowed to deliver normally and were sacrificed at weaning. The F2b animals were mated once and allo wed to raise their offspring to weaning when both parents and offspr ing were sacrificed.

Gross necropsies were performed on all parent animals. Offsping from the F1a, F2a, F2c and F3a generations were subject to gro ss necropsies at weaning. Tissues from 5 animals of each sex/dose of the F1b parents and the F3a generation were fi xed at necropsy. Tissue examination from the control and high dose groups included stomach, ile um, jejunum, colon, liver, spleen, hearth, lungs, adrenals, kidneys, urinary bladder, th yroid, ovaries, and ut erus or testes.

No treatment related effects on food cons umption, body weight, adult mortality, mating performance, pregnancy and fertility rates, gest ation length, offspring survival, weights and sex, litter survival, resorption rates, or necropsy findings. There were also no macroscopic or microscopic tissue abnormalities attributable to treatment. The NOAEL was 1000 mg/kg bw/day. (JECFA, 1986)

Mutagenicity

In a Salmonella/microsome assay, Fast Green FCF was determined to be non-mutagenic. Negative

results were also found in bact erial DNA repair tests. In another test, color-induced cell

transformation occurred in cult ured Fisher rat embryo cells at a concentration of 1μg/ml. Chromosome damage was also repor ted in an in vitro test using Chinese hamster ovary cells. (JECFA, 1986)

Neurotoxicity

No neurotoxicity studies were av ailable for review. However, signs of neurotoxicity were not observed in any of the studies that were submitte d and EPA concluded that a study is not required

at this time.

Immunotoxicity

No immunotoxicity studies were available for revi ew. However, signs of immunotoxicity were not observed in any of the studies that were submitted and EPA concluded that studies are not required

at this time.

Metabolism and Pharmacokinetics

The metabolism potential of Fast Green FCF was test ed in rats and dogs. Each species was given orally 200 mg of the color. In the rats, the urine and feces were collected for 36 hours. In the dogs, a bile fistula was made for bi le analysis. Almost all of th e color was excreted unchanged in

the feces of the rats and no color was found in the ur ine. The color did not exceed 5% of the given12 dose in the bile of the dogs. After feeding, the color was found in the bile of rats, but not in the

urine. It was concluded that the amount found in the bile prov ides an estimate of the amount absorbed through the gastrointe stinal tract. (JECFA, 1986)

C. Toxicity Endpoint Selection

The available toxicity study indica tes that FD&C Green No. 3 has a very low overall toxicity. The lowest NOAEL in the database is 1000 mg/kg bw /day. In the carcinogenicity study with an in utero phase, the effects on the pups (decrease d body weights and pup mortality) and kidney and thyroid toxicity in adults were observed at 5% in diet (equivale nt to 2500 mg/kg/day). Since these

effects were observed at 2.5 times the limit dose of 1000 mg/kg/day, there are low concerns for the hazard. Since no endpoint of concern was identif ied for the acute and chronic dietary exposure

assessments and short- and intermediate-term de rmal and inhalation exposure assessments, a quantitative risk assessment for FD &C Green No. 3 is not necessary.

V. FQPA Safety Factor Considerations

Section 408(b)(2)(c) of FFDCA provides that EPA sh all apply an additional tenfold (10X) margin of safety for infants and children in the case of threshold effects to account for prenatal and postnatal toxicity and the comp leteness of the database on to xicity and exposure unless EPA

determines based on reliable data that a different margin of safety will be safe for infants and children. This additional marg in 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 safety factor when reli able 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 n eeded for assessing risk to infants and children. The available toxicity studies suggest low toxi city of FD&C Green No. 3. The t oxicity database for FD&C Green

No. 3 contains an acute oral t oxicity study and chroni c toxicity studies, in cluding carcinogenicity, and reproductive toxicity studies. No reproductive or developmental toxic ity was observed in the 3-generation reproduction study. The database also contains mutagenicity studies, and metabolism data. There is no indication, base d upon the available data, that FD &C Green No. 3 is a neurotoxic

or immunotoxic chemical or results in increased qualitative or quant itative susceptibility in infants

or children. Based on this informa tion, 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.

VI. Exposure

A. Dietary Exposure13 Dietary exposure to FD&C Green No. 3 can occur from eating food that has come in contact with surfaces treated with pesticide fo rmulations containing this inert ingredient. Dietary exposure to FD&C Green No. 3 can also occur from eating f oods which contain FD&C Green NO. 3 as an ingredient. However, since an endpoint of concern for risk assessment was not identified, a quantitative dietary exposure assessment for FD&C Green No. 3 was not conducted.

i. Acute exposure . No adverse effects attributable to a single exposure of FD&C Green No. 3 were seen in the toxicity databa ses. Therefore, acute dietary ri sk assessment for FD&C Green No. 3 is not required.

ii. Chronic exposure. A chronic dietary exposure asse ssment was not conducted for FD&C Green No. 3 because no toxicological endpoint of concern was identified for chronic dietary exposure. The available studies/l iterature contained no evidence of a chronic dietary endpoint of

concern.

iii. Dietary exposure from drinking water: Dietary exposure from dr inking water to FD&C Green No. 3 can occur by drinking water that has been contaminated by run-off from a pesticide treated area, such as countertops. Since an e ndpoint for risk assessment was not identified, a quantitative dietary exposure assessment from drinking water for FD&C Green No. 3 was not

conducted.

iv. Cancer exposure. Based on the available toxicity data (carcinogenicity study in rats and mice) and lack of mutagenicity concerns, FD&C Green No. 3 is not expe cted to be carcinogenic. Therefore, a cancer dietary exposure assessment is not necessary to assess cancer risk.

B. Residential (Non-Occupational) 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, i ndoor pest control, termiticides, and flea and tick control on pets). The proposed use of FD&C Green No. 3 as a dye 40 CFR 180.940(a) is not expected to result in residential exposure to this chemical. Reside ntial exposure is possible based on other currently approved inert uses of this chemical.

However, since there are no toxicological effects of concern identified in the available database_, it is not necessary to conduct a ssessments of residentia l (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

i. Acute Aggregate Risk. There was no hazard attributable to a single exposure seen the in the toxicity database for the FD&C Green No. 3; therefore an acute aggregate risk assessment is not required

.14 ii. Short-term Aggregate risk

. Short-term aggregate exposure ta kes into account short-term residential exposure plus chronic exposure to food and water (considered to be a background exposure level). The requested use pattern for FD&C Green No. 3 could result in short-term residential exposure.

Since there are no oral, dermal or inhalation toxicological endpoints of concern identified in the available database, it is not necessary to aggreg ate dermal and inhalation exposures with estimated dietary exposures.

Residential Post-application Exposure

Assessments of residential post-application exposure are not necessa ry since there are no toxicological endpoints of concern iden tified in the available database.

VIII. Aggregate Risk Assessme nts and Risk Characterization

i. Acute Aggregate Risk. There was no hazard attributable to a single exposure seen the in the toxicity database for FD&C Green No. 3, therefore, an acute aggregate risk assessment is not required.

ii. Short-Term/Intermediate -Term Aggregate Risk. Short-term and intermediate-term aggregate risk assessments for FD&C Green No. 3 were not conducted. The assessments are not necessary since there are no dermal or inhalation t oxicological effects of concern identified in the

available database. There are no risks that exceed the Agency’s level of concern.

IX. Occupational Exposure

In the case of FD&C Green No. 3, the Agen cy has reviewed the av ailable toxicological

information. The data indicate that there ar e no adverse toxicologica l effects (endpoints of concern) identified in the available database, therefore 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 cons idering whether to establish, modi fy, or revoke a tolerance, the Agency consider “available information” conc erning the cumulative effects of a particular pesticide's residues and “other substances th at have a common mechanism of toxicity.”

EPA has not found FD&C Green No. 3 to share a common mechanism of toxicity with any other substances, and FD&C Green No. 3 does do not a ppear to produce a toxic metabolite produced by other substances. For the purposes of this tolera nce action, therefore, EP A has assumed that the

FD&C Green No. 3 does not have a common mechan ism of toxicity with other substances. For information regarding EPA's efforts to determin e which chemicals have a common mechanism of15 toxicity and to evaluate the cumulative effect s of such chemicals, see EPA's website at http://www.epa.gov/pesticides/cumulative

.

XI. Enviromental Fate

“Fast Green FCF's production and use as a colo r additive in foods, drugs and cosmetics may result in its release to the environment th rough various waste streams. If released to air, Fast Green FCF will exist solely in the particulate phase in the atmosphere because it is a salt and is nonvolatile. Particulate-phase Fast Green FCF will be removed from the atmosphere by wet or dry deposition. Fast Green FCF may be sus ceptible to direct photolysis by sunlight; its absorption maximum is 628 nm. If released to soil, Fast Green FCF is expected to be mobile; it is expected to exist almost entirely in anion form in the environment and anions generally do not adsorb more strongly to so ils containing organic carbon and clay than their neutral counterparts . Volatilization from mo ist soil surfaces is not expected to be an important fate proce ss because the compound exists as an anion and anions do not volatilize. If released into water, Fast Green FCF is not expected to adsorb to suspended solids and sediment based upon this compound's ionic nature in the environment. The concentrations of azo dye s with a phenolic OH and two sulfonic acid groups remain constant throughout activated sludge treatment and are therefore not expected to be biodegradable in water and so ils. Volatilization from water surfaces is not expected to be an important fate process based up on the ionic character of the compound. The potential for bioconcentrati on in aquatic organisms is lo w based on the ionic character of the compound. Hydrolysis is not expected to be an important environmental fate process since this compound lacks f unctional groups that hydrol yze under environmental conditions. Occupational exposur e to Fast Green FCF may o ccur through dermal contact with this compound at workplaces where Fast Green FCF is produced or used. Use data indicate that the gene ral population may be exposed to Fa st Green FCF via ingestion of food and drug products that contain this color additive, and dermal contact with cosmetics that use this compound.” (US NLM, TOXNET search)

XII. Ecological Effects

Ecological Structure-Activity Relationship M odel (ECOSAR) estimations indicate that FD&C Green No. 3 has potentially low toxicity to aquatic organisms (fish and invert ebrates) and to algae.

XIII. Determination of Safety

.

Taking into consideration all available inform ation on FD&C Green No. 3, EPA has determined that there is a reasonable certainty that no harm to any population subgroup will result from aggregate exposure to FD&C Green No. 3 under reas onable foreseeable circumstances. Therefore, the establishment of an exemp tion from tolerance under 40 CFR 180

. 940(a) for residues of FD&C Green No. 3 when used as an inert ingredient (d ye) in antimicrobial formulations, for use on food contact surfaces in public eating places, dair y processing equipment, and food processing equipment and utensils, is safe under FFDCA section 408.16 XIV. Environmental Justice

Potential areas of environmental ju stice concerns, to the extent possi ble, were considered in this human health risk assessment, in accordance w ith U.S. Executive Order 12898, "Federal Actions

to Address Environmental Just ice in Minority Populations an d Low-Income Populations," http://www.eh.doe.gov/oepa/ guidance/justice/eo12898.pdf ).

As a part of every pesticide risk assessment, OP P 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 patterns of that subgroup’s food and water consumption, and activities in and ar ound 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 Indivi duals (CSFII) and are used in pesticide risk assessments for all registered food uses of a pestic ide. These data are analyzed and categorized by subgroups based on age, season of the year, ethni c group, and region of the country. Additionally, OPP is able to assess dietary exposure to sma ller, specialized subgroups and exposure assessments are performed when conditions or circumstances warrant. Whenever appropriate, non-dietary xposures based on home use of pesticide products and associated risks for adult applicators and for toddlers, youths, and adults ente ring or playing on treated area s postapplication are evaluated. Further considerations are currently in developm ent as OPP has committed resources and expertise to the development of specialized software and m odels that consider exposure to bystanders and farm workers as well as lifestyle and traditio nal dietary patterns among specific subgroups.

XV. Human Studies

This assessment does not rely on an y studies in which human subjec ts were intentionally exposed to a pesticide.

XVI. International limits

In making its tolerance decisions, EPA seeks to harmonize U.S. tolerances with international standards whenever possible, consis tent with U.S. food safety standa rds and agricultur al practices.

EPA considers the international maximum resi due limits (MRLs) established by the Codex Alimentarius Commission (Codex), as requi red by FFDCA section 408(b)(4). The Codex Alimentarius is a joint United Nation Food and Agriculture Organization/World Health Organization food standards prog ram, and it is recognized as an international food safety standards-setting organization in tr ade 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 explain the re asons for departing from the Codex level. The Codex has not

established a MRL for FD&C Green No. 3.17 References

ECOSAR, 2000. Ecological Structure Activity Rela tionships Version 0.99g. Environmental Protection Agency. http://www .epa. gov I oppt!N ewchems/21 ecosar.htm

ECOTOX. 2002 U.S. Envir onmental Protection Agency, 2002 ECOTOX User Guide: ECOTOXicology Database System. Versi on 3.0 http://www.epa .gov/ecotox/ Search terms: CAS Reg Nos: 2353-45-9.

FDA, 2011, Code of Federal Regulations , Title 21, Volume 1, 21 CFR 74.203

HSDB. 2004. Hazardous Substance Data Bank (HSD B). Online Scientific Search Engine, National Library of Medicine, Na tional Institutes of Health. http://toxnet.nlm.nih. gov Icgibin/sis/htmlgen?HSDB

Search terms: FD&C Green No. 3 (CAS RN 2353-45-9); August 18, 2013

WHO Food Additive Series 21, Prepared By: The Joint FAO/WHO Expert Committee on Food Additives (JECFA), 1987, FAST GREEN FCF, In ternational Program Chemical Safety (IPCS)

INCHEM

WHO Food Additive Series 20, Prepared By: The Joint FAO/WHO Expert Committee on Food Additives (JECFA), 1986, FAST GREEN FCF, In ternational Program Chemical Safety (IPCS)

INCHEM

WHO Food Additive Series 16, Prepared By: The Joint FAO/WHO Expert Committee on Food Additives (JECFA), 1982, FAST GREEN FCF, In ternational Program Chemical Safety (IPCS)

INCHEM

WHO, 30th Report of the Joint Expert Committ ee on Food Additives, 1987, Evaluation of certain food additives and contaminants, Technical Report Series, 751, Page 24. Page 18