May 9, 2014

MEMORANDUM

SUBJECT: Decision Document for Petition Number 1E7843: FD&C Red No. 40 (CAS Reg. 

No. 25956-17-6); Human Health Risk Assessment and Ecological Effects 

Assessment for Proposed Exemption from the Requirement of a Tolerance When 

Used as an Inert Ingredient in Pesticide Formulations.

PC Code: 911363

DP Barcode: N/A

Decision No.: N/A

Regulatory Action: Inert Tolerance Exemption

40 CFR 180.940(a)

Petition No.: 1E7843

All CAS Reg. Nos: 25956-17-6

Inert Tracking No N/A

FROM: Roger Chesser, Biologist

Inert Ingredient Assessment Branch (IIAB)

Registration Division (7505P)

THRU: Kerry Leifer, Team Leader

Inert Ingredient Assessment Branch (IIAB)

Registration Division (7505P)

TO: P.V. Shah, Chief

Inert Ingredient Assessment Branch (IIAB)

Registration Division (7505P)

UNITED STATES ENVIRONMENTAL PROTECTION AGENCY

WASHINGTON, D.C. 20460

OFFICE OF CHEMICAL SAFETY 

AND POLLUTION PREVENTION
2

Table of Contents

Executive Summary

I. Background...........................................................................................................................................................4

II. Inert Ingredient Profile..........................................................................................................................................5

III. Human Health Assessment...............................................................................................................................6

A. Summary of Toxicity Data ................................................................................................................6

B. Toxicity Endpoint Selection...............................................................................................................13

C. Special Consideration for Infants and Children .................................................................................13

IV. Exposure Assessment ............................................................................................................................................13

V. Aggregate Risk and Determination of Safety .........................................................................................................13

Acute Dietary Risk..................................................................................................................................13

Chronic Dietary Risk ..............................................................................................................................14

Aggregate Cancer Risk for U.S. Population ...........................................................................................14

Short-Term Aggregate Risk....................................................................................................................14

Intermediate-Term Aggregate Risk ........................................................................................................14

Determination of Safety..........................................................................................................................14

VI. Occupational Exposure / Risk Pathway.................................................................................................................14

Occupational Handler Risk .....................................................................................................................15

Occupational Post-Application Risk.......................................................................................................15

 Exposure Scenarios.........................................................................................................................15

VII. Cumulative exposure............................................................................................................................................16

VIII. Environmental Justice Statement........................................................................................................................16

IX. Environmental Fate Considerations.......................................................................................................................17

X. Ecotoxicity..............................................................................................................................................................17

XI. Risk Characterization ............................................................................................................................................17

References ...................................................................................................................................................................18
3

Executive Summary

Diversey, Inc. (8310 16th Street, MS 707, Sturtevant WI 53177-1964) submitted a 

petition (1E7843) to establish an exemption from requirement of a tolerance, pursuant to section 

408 of the Federal Food, Drug, and Cosmetic Act, for the use of FD&C Red No. 40 as an 

antimicrobial pesticide formulation inert ingredient (colorant) in food contact surface sanitizer 

products under 40 CFR §180.940 (a) at a maximum level in the end-use concentration of 20 parts 

per million (ppm). The majority of the data in this risk assessment decision document have been 

taken from the tolerance reassessment document for FD&C Red No. 40 dated December 21, 

2004 (MRID 48779801) evaluating its use as an inert ingredient in pesticide formulations 

applied to growing crops and raw agricultural commodities after harvest (40 CFR 180.910), and 

to growing crops only (pre-harvest seed treatment) (40 CFR 180.920).

 The reassessment document also summarized available information on the use, 

physical/chemical properties, toxicological effects, exposure profile, environmental fate, and 

ecotoxicity of FD&C Red No. 40, including data considered by the Joint Expert Committee on 

Food Additives of the Food and Agriculture Organization/World Health Organization 

(FAO/WHO) Expert Committee on Food Additives (JECFA) (MRIDs 48779804 and 48779805)

as well as the European Food Safety Authority (EFSA) (MRID 48779803). As summarized in 

that reassessment document, all toxicological studies were conducted at the limit dose or close to 

limit dose. No adverse toxicological effects were detected at these doses. A residential risk 

assessment was not performed since there are no residential uses. No endpoint of concern was 

identified in the database. 

FD&C Red No. 40 is a FDA permanently listed color additive used in food, drugs and 

cosmetics, including drugs and cosmetics for eye area. The FDA’s color additive evaluation 

included the consideration of an extensive set of toxicological data on FD&C Red No. 40. FDA 

concluded that this colorant was safe and established a maximum acceptable daily intake for 

FD&C Red No. 40 of 7 mg/kg-bw/day. Similarly, JECFA and SCF have evaluated FD&C Red 

No. 40 for the purpose of establishing estimates of acceptable daily intake (ADI) as a food 

additive. 

The JECFA and SCF evaluations of FD&C Red No. 40 included reviews of an extensive 

set of toxicological data including genotoxicity, chronic toxicity /carcinogenicity and 

reproductive and developmental toxicity. Both evaluations concluded that the available data 

demonstrated that no adverse effects were seen in studies at limit dose levels. No hazard was 

identified in any of the data evaluated by JECFA for FD&C Red No. 40 at the limit dose of 1000 

mg/kg/day to either parental animals or their offspring. Acute oral toxicity was low based on 

studies with rats (LD50 > 10000 mg/kg bw). Based on other acute oral toxicity studies with 

rabbits and dogs, JECFA set the the LD50 values at > 10000 mg/kg bw and 5000 mg/kg bw, 

respectively. The JECFA evaluation resulted in the establishment of an ADI of 7.0 mg/kg/day 
4

based on a chronic oral toxicity study in rats in which the no-observed-adverse-effect level 

(NOAEL) was 695 mg/kg/day (highest dose tested) with a safety factor of 100. (JECFA 

evaluation based upon Serota et al, 1977, as cited in JEFCA, 1980) The JECFA and SCF 

evaluations of FD&C Red No. 40 established ADIs of 7.0 mg/kg/day based upon a chronic 

dietary toxicity study in rats in which the no-observed-adverse-effect level (NOAEL) was 750 

mg/kg/day (highest dose tested) with a safety factor of 100. 

More recently, the European Food Safety Authority (EFSA) has reevaluated FD&C Red 

No. 40 and concluded that the present database does not give reason to revise the ADI of 7.0

mg/kg bw/day. As a result of these extensive evaluations of FD&C Red No. 40, in which either 

no adverse effects were noted, or the effects of single or repeated dosing were observed only at 

levels beyond the respective limit doses, EPA has utilized a qualitative approach to assessing 

human health risks from exposure to FD&C Red No. 40. No hazard endpoint of concern was 

identified for the acute and chronic dietary assessment (food and drinking water), or for the 

short, intermediate, and long term residential assessments (via all exposure routes), therefore, 

acute and chronic dietary and short-, intermediate, and long-term residential exposure 

assessments were not performed. 

Thus, due to its low potential hazard and the lack of a hazard endpoint, it has been 

determined that a quantitative risk assessment using safety factors applied to a point of departure 

protective of an identified hazard endpoint is not appropriate for FD&C Red No. 40 and an 

FQPA safety factor is not needed to protect the safety of infants and children. 

Residues of concern are not anticipated for dietary exposure (food and drinking water) or 

for residential exposure from the use of FD&C Red No. 40 for the proposed use pattern as an 

inert ingredient in pesticide products. EPA expects aggregate exposure to FD&C Red No. 40 to 

pose no appreciable dietary risk given that the data on FD&C Red No. 40 show a lack of any 

systemic toxicity at limit dose levels and a lack of any apparent developmental effects (SIDS). 

Taking into consideration all available information, EPA concludes that there is a 

reasonable certainty of no harm to the general population or to infants and children from 

aggregate exposure to FD&C Red No. 40 when used as an inert ingredient in pesticide 

formulations. Therefore, the establishment of an exemption from the requirement of a tolerance 

under 40 CFR 180.940 (a) for FD&C Red No. 40 can be considered safe under section 408 of the 

FFDCA. 

I. Background

Diversey, Inc. (8310 16th Street, MS 707, Sturtevant WI 53177-1964) is requesting EPA to 

establish a tolerance exemption for FD&C Red No. 40, CAS #: 25956-17-6 for use as an 
5

antimicrobial pesticide formulation inert ingredient (colorant) in no-rinse, food contact surface 

sanitizer products under 40 CFR § 180.940(a) at a maximum level in the end-use concentration 

of 20 parts per million (ppm).

In addition to the EPA approved uses as an inert ingredient in pesticides, FD&C Red No. 40 is 

approved by the Food and Drug Administration (FDA) as a color additive in food products as 

specified at 21 CFR §74.340, for use in drugs at 21 CFR §74.1340 and for use in cosmetics at 21 

CFR §74.2340. It is permanently listed as safe for general use in these products. 

EPA completed reassessment of the current established tolerance exemption FD&C Red 

No. 40 in December 2004. The Agency concluded that taking into consideration all available 

information, including the extensive toxicology database in which no adverse effects were noted, 

the low toxicity, the lack of risk concerns from usage as color additives, and the low exposures 

likely to result from uses as pesticide active ingredients, that there was reasonable certainty of no 

harm to any population subgroup from aggregate exposure to FD&C Red No. 40 , considering 

dietary and all other non-occupational exposures. 

The European Food Safety Authority (EFSA); Panel on Food Additives and Nutrient 

Sources added to Food also reevaluated the safety of FD&C Red No. 40 as a food coloring 

additive and published their scientific opinion in 2009. They concluded that no revisions to the 

current established ADI of 7 mg/kg/bw were needed. 

Taking into consideration all available information, EPA concludes that there is a 

reasonable certainty of no harm to the general population or to infants and children from 

aggregate exposure to FD&C Red No. 40 when used as an inert ingredient in pesticide 

formulations. Therefore, the establishment of an exemption from the requirement of a tolerance 

under 40 CFR 180.940(a) for FD&C Red No. 40 can be considered safe under section 408 of the 

FFDCA. 

II. Inert Ingredient Profile

A. Summary of Uses

FD&C Red No. 40 is used as an inert ingredient in pesticide formulations that are intended for 

application on growing crops or on raw agricultural commodities after harvest (40 CFR 

180.910). The colorant cannot be used above 0.002% (20 ppm) by weight in the formulation 

and must meet the certification requirements specified in 21 CFR 74.340. FD&C Red No. 40 is 

similarly approved for use as a dye or coloring agent for seed treatment with a maximum 

concentration of 2% by weight in the pesticide formulation (40 CFR 180.920).
6

B. Physical and Chemical Properties

Some of the physical and chemical characteristics of FD&C Red No. 40, along with its structure 

and nomenclature follow:

Source ChemIDplus Lite

Chemical Name: FD&C Red No. 40

CAS No.: 25956-17-6

Molecular Formula: C18-H16-N2-O8-S2.2Na 

Structure: 

Source SRC, 2004:

Molecular Weight: 496.427

Water solubility: 2.25 x 105 mg/L at 25° C (M)

Octanol-water Partition Coefficient (Kow) log Kow= -0.550 (E)

Source: (MPBPVPWIN EPI Suite, 2000 as cited by IACM, 2003)

MP: 350 C 

Vapor pressure: 1.25 x 105 mm Hg @ 25° C

Henry’s Law Constant: 1 x 10-15 atm-m3/mole @ 25°C (estimated)

Based on the above data, FD&C Red No. 40 has the following physicochemical 

profile:

 Water soluble

 Nonvolatile

III. Human Health Assessment

A. Summary of Toxicity Data
7

The European Food Safety Authority (EFSA) has conducted the most recent (2009) full 

review of the toxicology of FD&C Red No. 40. This document relied heavily on the earlier 

reviews conducted by the Joint FAO/WHO Expert Committee on Food Additives (JECFA) 

which set the current, permanent Acceptable Daily Intake (ADI) of 07 mg/kg-bw/day.(JECFA, 

1981). 

Acute oral toxicity. No compound related effects were observed when FD&C Red No. 40

(Allura Red AC) was given to rats by gavage at doses varying from 215 to 10000 mg/kg. Based on

these studies, JECFA set the LD50 for rats at 10000 mg/kg-bw. Other studies in rabbits, dogs and mice 

produced LD50 values of 10,000, 5000 and >2000 mg/kg, respectively. Overall, the EFSA 

concluded that “the acute oral toxicity of Allura Red AC (FD&C Red No. 40) islow.”

Dermal Toxicity Tests for dermal irritation in rabbits on intact and abraded skin 

showed no gross irritation at levels of .316, 1,03,16 abd 10 g/kg b/w of Allura Red. Daily 

applications of the color at rates of (JECFA, 1980 )

Inhalation Toxicity. No studies available in the database.

Primary Eye Irritation. No studies available in the database.

Table 1. Acute Toxicity Profile of FD&C Red No. 40 

Study Type Results

Toxicity 

Category MRIDs # 

/Reference

Acute Oral - rat LD50 > 10,000 mg/kg 

bw 

IV 48779804; JECFA

1981

Acute Oral - rabbit LD50 > 10,000 mg/kg 

bw 

IV 48779804; JECFA

1981

Acute Oral - dog LD50 = 5,000 mg/kg 

bw 

III 48779804; JECFA

1981

Acute Oral - mice LD50 > 2,000 mg/kg 

bw 

IV 48779804; JECFA

1981

Primary Skin Irritation -

rabbit

Minimal to mild 

irritation 

IV 48779804; JECFA

1981
8

Genotoxicity. EFSA cited reviews by the European Scientific Committee for Food 

(SCA), the JECFA and by TemaNord, to conclude that FD&C Red No. 40 was determined to 

be non-mutagenic in a variety of in vitro and in vivo based mutagenicity studies. This 

determination is consistent with EPA’s conclusion in the 2004 Tolerance Reassessment for 

FD&C Red No. 40 (EPA, 2004).

In their 2009 review, EFSA cited a study from 2001 conducted on FD&C Red No. 40 

using the Comet assay. The study showed an increase in migration of nuclear DNA in the 

glandular stomach and the colon of male mice exposed to doses of FD&C Red No. 40 of 100 

mg/kg-bw and 10 mg/kg-bw respectively. Since histopathology of the effected organs 

appeared negative, the study authors concluded that observed effect was not likely to be related 

to cytotoxicity. In light of the negative in vivo carcinogenicity studies conducted on FD&C 

Red No. 40 (see below), EFSA concluded that the significance of the Comet assay results was 

uncertain.

Reproductive and developmental toxicity. Relevant reproductive and developmental 

toxicity studies cited in the EFSA document are summarized in Table 1. Blackmore, 1969, 

observed slight growth suppression in F1 and F2 pups exposed to 5.19% of FD&C Red No. 40 

in the diet during a multi-generation type reproduction study in rats. This dose was calculated 

to be equivalent to 2595 mg/kg/day. The fetal No Adverse Effect Level (NOAEL) was 1.39% 

in the diet; equivalent to 695 mg/kg/day. The maternal NOEAL was 5.19% in the diet, which 

was equivalent to 2595 mg/kg/day. No other significant anomalies were detected in growth, 

litter size, pup weight, gross pathology, implantation sites, resorption sites, live fetuses indices, 

appearance, anatomy and structure were observed. The ADI of 7 mg/kg-bw/day for FD&C 

Red No. 40 was partially based on this study.

Teratology studies in rats and rabbits showed no evidence of adverse effects at 200 mg/kgbw/day (gestation day (g.d.) 0-19) and 700 mg/kg/day (g.d. 6-18), respectively.

Reproductive toxicity endpoints were also monitored in three long-term studies.

In a lifetime study in mice, the animals were fed up to 5.19% of FD&C Red No. 40 in 

the diet. Animals were also exposed throughout gestation and lactation. At the highest dose 

level, lower body weights and effects on organ weight and organ/body weight ratios were 

noted but considered to be “inconsequential.” (Serota et al., 1977a).

In a nearly identical lifetime study with larger group numbers (100/sex/group versus

50/sex/group in the original study), no compound related adverse effects were observed. The 

NOAEL’s in both studies were determined to be 5.19% FD&C Red No. 40 in the diet, which was 
9

calculated to be equivalent to 7300 and 8300 mg/kg- bw/day in male and female mice, 

respectively. (Reno et al., 1978)

Dietary administration of 5.19% FD&C Red No. 40 in rats was also studied (Serota et 

al., 1977b). Administration started throughout parental breeding, gestation and lactation and 

continued for 118 and 121 weeks in male and female rats, respectively. For the female rats in 

the study, mean body weights and growth rates were significantly lower in the high dose group 

relative to controls. No other significant adverse effects were reported. The NOAEL was 

5.19% (2821 mg/kg-bw/day) in male rats and 1.3% (901 mg/kg-bw/day) in female rats.

A review conducted by TemaNord, 2002, discussed four additional 

reproductive/developmental toxicity studies.

Rats (group number not reported) were exposed to up to 10% of FD&C Red No. 40 in 

the diet (calculated doses of 0, 1250, 2500 and 5000 mg/kg-bw/day). Litter mortality was 

increased between 22-24 days of age at a concentration of 10% in the diet. Significantly 

decreased running wheel activity was observed in all exposed groups. Increased open-field 

rearing was observed in the two highest dose groups. The Low Adverse Effect Level (LOAEL) 

was determined to be 1250 mg/kg-bw/day; a NOAEL could not be determined from this study 

(Voorhees, et al., 1983).

In a mouse study, animals (10/sex/group) were exposed to up to 1.68% FD&C Red 

No. 40 in the diet over two generations (equivalent to 2400 mg/kg-bw/day). The study 

showed no significant, dose-related adverse effects on reproduction or

neurobehavioral endpoints following exposure to FD&C Red No. 40 (Tanaka, 1994).

In rats (group number not reported) dosed with FD&C Red No. 40 up to 0.7% in 

drinking water (equivalent to 939 mg/kg-bw/day) on g.d. 0-20, a significant increase in the 

incidence of fetuses with reduced ossification of the hyoid was observed at the highest dose 

level. No other fetal malformations were observed. The NOAEL from this study was 

determined to be 546 mg/kg-bw/day (Collins et al., 1989a).

In rats dosed with FD&C Red No. 40 on g.d. 0-19 (group number not reported), no 

significant, dose-related effects were observed at doses up to 1000 mg/kg/day (Collins et al., 

1989b).

Subchronic toxicity. FD&C Red No. 40 was studied in rats, dogs and pigs in the 

following short-term studies.

In rats, groups of 20 animals (10 males and 10 females per group) were fed diets 

containing 0-2595 mg/kg-bw/day) for six weeks. The NOAEL was determined to be at least 

2595 mg/kg-bw/day.
10

Both of the dog studies were determined to be too limited to derive a NOAEL.

The pig study appeared to be a non-standard design. Groups of 4 animals (2 males 

and 2 females per group) were dosed with 1000 mg/kg-bw/day of FD&C Red No. 40 for 

21 days. The dose was increased to 1500 mg/kg-bw/day for an additional 54 days. No 

compound related effects were reported.

Chronic toxicity. The chronic toxicity/carcinogenicity of FD&C Red No. 40 was 

studied in 5 key studies. These are summarized in Table 2.

In general, exposure to FD&C Red No. 40 did not result in significant adverse effects. 

In one mouse study, higher rates of early onset lymphoma were observed in animals exposed 

to FD&C Red No. 40 (Serota et al., 1977a). However, this observation was not statistically 

significant in this study and could not be replicated in a subsequent study that used a higher 

number of animals per group (100 versus 50) and an additional control group. In general, the 

only significant effects reported in these studies appeared to be growth suppression or body 

weight reductions at high dietary doses used in these studies.

Metabolism and Pharmacokinetics. The applicable studies were summarized in 

JECFA, 1980. In rats fed 5.19% FD&C Red No. 40 in the diet, only 0.1% and 29% of the 

unmetabolized dye was found to be excreted in the urine and feces, respectively. Using 35S 

labeled FD&C Red No. 40, in dogs, 92-95% of the recovered radioactivity appeared in the 

feces and between 2.7 and 3.6% in the urine within 72 hours of administration. In rats, 76-

92% of the radioactivity was recovered in the feces and 5.7-19.8% in the urine within the 

same timeframe. Significant retention of the dye was noted in the intestinal contents and in 

the washed intestines of both species. The total carcass and viscera of both species retained 

only 0.4% of the total administered dose.

FD&C Red No. 40 is absorbed to a limited extent and the major route of excretion is 

through metabolic by-products eliminated through the feces. Urinary excretion is negligible.

Metabolite toxicology. Cresidine-4-sulfonic acid was identified as the major metabolite 

in the urine and feces of both rats and dogs along with the parent compound and two unknown 

metabolites. In the dog, an additional unknown metabolite was observed in the feces and 

possibly the urine. Metabolism in the GI tract may occur through the reduction of the azo 

group in the parent compound.

EFSA, 2009, reported that FD&C Red No. 40 was able to inhibit aromatase activity 

in an in vitro rat ovarian microsome system with an IC50 of 24M. Aromatase catalyzes 

the conversion of androgens to estrogens and presents a possible target for endocrine 
11

disrupting chemicals. EFSA concluded; however, that these data did not trigger a concern 

for FD&C Red No. 40 since the chemical has limited systemic bioavailability and that 

there was no evidence of related adverse effects in the various reproduction studies 

conducted on the chemical.

Table 1: Summary of Reproductive and Developmental Toxicity Studies Conducted on FD&C Red No. 40 base

Reference Study Type Species Duration Dose Summary 

Blackmore et 

al., 1969

Reproduction 

(multigeneration)

Rat

10 male

10 female

Mating 

occurred 

after 27 

weeks of 

exposure

0, 0.37, 0.72, 1.39 or 

5.19 % in the diet. 

Equivalent to 0, 185, 

360, 695 and 2595 

mg/kg-bw/day as 

recalculated by 

Borzelleca et al., 

1991.)

Fertility indices for both filial generations 

were decreased in both treated and control 

groups. Slight growth suppression was 

observed in high dose pups. No other 

anomalies in regards to fertility, fetal 

development, teratology or pathology were 

observed. 

Collins et al., 

1974

Teratology Rat Gestation 

Day

0-19

0, 15,30, 100 and 200 

mg/kg-bw/day

No compound related effects were observed. 

Reno, 1974 Teratology Rabbit Gestation 

Day

6-18

0, 200 and 700 

mg/kg-bw/day

No compound related effects were observed. 

Serota et al., 

1977a; 

JECFA, 1980

Chronic Mouse

50/sex/

group

Lifetime 0, 0.37, 1.39 and 

5.19% in the diet 

(equivalent to 0, 507, 

1877 and 7422 

mg/kg-bw/day in 

male mice and 0, 

577, 2043 and 8304 

mg/kg-bw/day in 

female mice- as 

recalculated by 

Borzelleca et al., 

1991). 

Animals were exposed through gestation 

and lactation. At the highest dose levels, 

decreased body weight, organ weights and 

effects on organ/body weight were 

observed. No significant compound related 

effects were observed.

Reno, et al., 

1978. Later 

published in 

Borzelleca, 

1991

Chronic Mouse

100/sex/

group

Two 

negative 

control 

groups

Lifetime 0, 0.37, 1.39 and 

5.19% in the diet 

(equivalent to 0, 492, 

1821 and 7318

mg/kg-bw/day in 

male mice and 0, 

526, 2057 and 8356

mg/kg-bw/day in 

female mice- as 

recalculated by 

Borzelleca et al., 

1991). 

Sporadic changes in clinical signs, body 

weight, adrenal weights (both absolute and 

relative), food consumption and survival 

rates were observed. None of the changes 

were significant or followed a dose-response 

relationship.
12

Serota et al., 

1977b; later 

published as 

Borzelleca et 

al., 1989)

Chronic with 

breeding

Rat

50/sex/

group

Lifetime118-121 

weeks

0, 0.37, 1.39 and 

5.19% Allura Red 

AC in the diet 

(equivalent to 180, 

701 and 2829 mg/kgbw/day for male and 

0, 228, 901 and 3604 

mg/kg-bw/day for 

female rats- as 

recalculated by 

Borzelleca et al., 

1989)

Administration started throughout parental 

breeding, gestation and lactation, and lasted 

for 118 and 121 weeks for male and female 

rats, respectively.

In females, mean body weights and growth 

rates were significantly decreased (-12.5%) 

at 3604-m/kg bw/day. 

Some abnormalities in gross pathological 

and histopathological appearance of the 

kidney were observed in groups fed FD&C 

Red No. 40. There was no observed doseresponse relationship. 

No compound-related adverse effects were 

observed, except for a reduction in body 

weight in high-dosed females at the end of 

the study. Borzelleca et al. (1989).

Voorhees et 

al., 1983-

from Tema 

Nord, 2004.

Multigeneration-2Rat

Group 

number 

not 

recorded.

Not 

specified.

0, 2.5, 5 or 10% 

FD&C Red No. 40 in 

the diet (equivalent to 

0, 1250, 2500 and 

5000 mg/kg-bw/day, 

respectively). 

Reproductive success (dams producing 

litters) was reduced at all dose levels but 

effect was not dose-related. Litter mortality 

was increased between 22-24 days of age at 

a concentration of 10% in the diet. 

Significantly decreased running wheel 

activity as observed in all exposed groups. 

Increased open-field rearing was observed in 

the two highest dose groups. (Voorhees et 

al., 1983). 

Tanaka, 1994 Multi-generation 

(2)

Mouse

10/sex/

group

0, 0.42, 0.84 or 

1.68% in the diet 

(equivalent to 0, 600, 

1200 and 2400 

mg/kg-bw/day). 

Animals showed no significant, dose-related 

adverse effects on reproduction or 

neurobehavioral endpoints following 

exposure to FD&C Red No. 40. 

Collins et al., 

1989a

Teratology Rat

Group 

number 

not 

recorded.

G.d. 0-20 0, 0.2, 0.4 or 0.7% in 

drinking water 

(equivalent to 0, 241, 

546 and 939 mg/kgbw/day). 

A significant increase in the incidence of 

fetuses with reduced ossification of the 

hyoid was observed at the highest dose 

level. No other fetal or maternal effects 

were observed. 

Collins et al., 

1989b

Teratology Rat G.d. 0-19 0, 30, 75, 150, 300, 

600 and 1000 mg/kgbw/day by gavage.

No significant, dose-related effects were 

observed at doses up to 1000 mg/kg-bw/day. 
13

B. Toxicity Endpoint Selection

No toxicological endpoints have been identified for FD&C Red No. 40 due to no adverse 

observable effects seen in toxicity studies at or above limit dose levels. 

C. Special Consideration for Infants and Children

At this time, there is no concern for potential sensitivity to infants and children resulting 

from exposures to FD&C Red No. 40. There is no reported quantitative or qualitative evidence 

of increased susceptibility of rat fetuses to in utero exposure to FD&C Red No. 40 in 

developmental toxicity studies in rats. No quantitative or qualitative evidence of increased 

susceptibility has been reported following the pre/postnatal exposure to rats in 2-generation 

reproduction toxicity studies in rats. Given the lack of adverse toxicological effects at limit dose 

levels, a safety factor analysis has not been used to assess the risk. For these reasons the 

additional tenfold safety factor is unnecessary.

IV. Exposure Assessment

No hazard endpoint of concern was identified for the acute and chronic dietary 

assessment (food and drinking water), or for the short, intermediate, and long term residential 

assessments (via all exposure routes), therefore, acute and chronic dietary and short-, 

intermediate, and long-term residential exposure assessments were not performed for FD&C Red 

No. 40. In addition, exposures to FD&C Red No 40 resulting from its use as an inert ingredient 

in pesticide formulations would be orders of magnitude less than the ADI’s established for 

FD&C Red No. 40 by FDA/JECFA/SCF.

V. Aggregate Risk and Determination of Safety

EPA determines whether acute and chronic pesticide exposures are safe by comparing 

aggregate exposure estimates to the aPAD and cPAD. The aPAD and cPAD represent the 

highest safe exposures, taking into account all appropriate safety factors (SFs). EPA calculates 

the aPAD and cPAD by dividing the POD by all applicable UFs. For linear cancer risks, EPA 

calculates the probability of additional cancer cases given the estimated aggregate exposure. 

Short-, intermediate, and chronic-term risks are evaluated by comparing the estimated aggregate 

food, water, and residential exposure to the POD. The MOE is the product of all applicable UFs 

and should not be exceeded. 

Acute Dietary Risk 

An acute aggregate risk assessment takes into account exposure estimates from acute dietary 

consumption of food and drinking water using the exposure assumptions discussed in section IV 
14

for acute exposure and the use limitations in pesticide formulations. There was no hazard 

attributable to a single exposure seen in the toxicity database for FD&C Red No. 40; therefore an 

acute aggregate risk assessment is not required. 

Chronic Dietary Risk

A chronic aggregate risk assessment takes into account exposure estimates from chronic dietary 

consumption of food and drinking water using the exposure assumptions discussed in section IV 

for chronic exposure and the use limitations in pesticide formulations. There was no hazard 

attributable to chronic exposures in the toxicity database for FD&C Red No. 40; therefore a

chronic aggregate risk assessment is not required.

Aggregate Cancer Risk for U.S. Population

The Agency has not identified any concerns for carcinogenicity relating to FD&C Red No. 40; 

therefore, a cancer dietary exposure assessment was not performed and an aggregate risk 

assessment is not a concern.

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 background exposure level). FD&C Red No. 40

is used as an inert ingredient in pesticide products that are currently registered for uses that could 

result in short-term residential exposure however no adverse effects were attributable to shortterm exposure to FD&C Red No. 40 were seen in the toxicity database; therefore, a short-term 

aggregate risk assessment is not required. 

Intermediate-Term Aggregate Risk

Intermediate-term aggregate exposure takes into account intermediate-term residential exposure 

plus chronic exposure to food and water (considered to be a background exposure level). FD&C 

Red No. 40 is used as an inert ingredient in pesticide products that are currently registered for 

uses that could result in intermediate-term residential exposure however no adverse effects were 

attributable to intermediate-term exposure to FD&C Red No. 40 were seen in the toxicity 

database; therefore, an intermediate-term aggregate risk assessment is not required. 

Determination of Safety

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 FD&C Red No. 40.

VI. Occupational Exposure / Risk Pathway
15

The representative occupational scenarios selected by the Agency for assessment are evaluated 

based on likely maximum application rates for products which may contain the inert ingredient 

for the short-term exposure assessment, and average application rates for products likely to 

contain the inert ingredient for the intermediate- and long-term exposure durations. Active 

ingredient application rates are corrected for the maximum amount of inert ingredient likely to be 

in the final formulations to determine exposure and risk from exposure to the inert ingredient by 

fungicide, insecticide or herbicide. 

RD traditionally considers a level of concern (LOC) for these risk assessments to be an MOE of 

100 based on the standard 10X inter and 10X intra species extrapolation safety factors and 

additional uncertainty in the database.

Occupational Handler Risk

 Exposure Scenarios

Due to the absence of dermal and inhalation toxicity endpoints, occupational handler risks are 

not of concern. Therefore an occupational handler risk assessment was not performed.

Occupational Post-Application Risk

 Exposure Scenarios

RD uses the term post-application to describe exposures that occur when individuals are present 

in an environment that has been previously treated with a pesticide (also referred to as re-entry 

exposure). Such exposures may occur when workers enter previously treated areas to perform 

job functions, including activities related to crop production, such as scouting for pests or 

harvesting. Post-application exposure levels vary over time and depend on such things as the 

type of activity, the nature of the crop or target that was treated, the type of pesticide application, 

and the chemical’s degradation properties. In addition, the timing of pesticide applications, 

relative to harvest activities, can greatly reduce the potential for post-application exposure.

Inhalation exposures are not typically calculated for occupational post-application scenarios 

because inhalation exposures generally account for a negligible percentage of the overall body 

burden for most pesticide chemicals. 

Due to the absence of dermal and inhalation toxicity endpoints, occupational post-application 

risks are not of concern. Therefore an occupational post-application risk assessment was not 

performed.
16

VII. 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 Red No. 40 to share a common mechanism of toxicity with any other 

substances, and that FD&C Red No. 40 does not appear to produce a toxic metabolite produced 

by other substances. For the purposes of this tolerance action, therefore, EPA has assumed that 

FD&C Red No. 40 does 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 and to evaluate the cumulative effects of such chemicals, see EPA's website at 

http://www.epa.gov/pesticides/cumulative.

VIII. Environmental Justice Statement

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 Low-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, RD 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 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 Intakes 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 risks 

for adult applicators and for toddlers, youths, and adults entering or playing on treated areas 

post-application are evaluated. Further considerations are currently in development as OPP has 

committed resources and expertise to the development of specialized software and models that 

consider exposure to bystanders and farm workers as well as lifestyle and traditional dietary 

patterns among specific subgroups.
17

IX. Environmental Fate Considerations

FD&C Red No. 40 is water soluble, mobile and not readily biodegradable. It is 

essentially nonvolatile; volatilization from soils and water is not likely to be a transport process 

in the environment.

EPA reviewed the environmental impact from use of FD&C Red No. 40 in agricultural 

pesticides. In agricultural use, the products are discharged directly into the environment without 

treatment. Based on their review, EPA concluded that “modeling results indicate that expected 

potential risks to aquatic organisms and soil and sediment dwelling organisms resulting from the 

use of FD&C Red No. 40 as an inert ingredient in pesticide formulations is low.” (EPA, 2004)

No-rinse food contact sanitizing solutions are typically indoor use products. These 

products are used at commercial facilities where there is no direct discharge to the environment 

during recommended use. The used solutions are drained into the sanitary sewer of the facility, 

where they are treated in accordance with permits issued under the Clean Water Act. After 

leaving the food processing facility, the concentration of FD&C Red No. 40 in the waste-water 

will be reduced further by dilution with other waste streams and by processing at the waste water 

treatment facility. Therefore, the ultimate concentration of FD&C Red No. 40 discharged by a 

waste-water treatment facility to the environment would be negligible

X. Ecotoxicity

There are no available aquatic toxicity studies on FD&C Red No. 40 (ECOTOX, 2002), 

however, based on estimates of toxicity using physical and chemical property data, FD&C Red 

No. 40 would be of low concern for toxicity to fish, algae, and aquatic invertebrates as well as 

terrestrial/benthic organisms (ECOSAR, 2000).

XI. Risk Characterization

No toxicological endpoints have been identified for FD&C Red No. 40 due to no adverse 

observable effects seen in toxicity studies at doses at or above limit dose levels .

The toxicity database for FD&C Red No. 40 is adequate for FQPA assessment and the potential 

exposure is adequately characterized given the low toxicity of the chemical. No hazard was 

identified and there is no residual uncertainty regarding prenatal and/or postnatal toxicity. No 

acute or subchronic neurotoxicity studies are available, but there were no clinical signs of 

neurotoxicity or any systemic toxicity observed in the available database at doses up to and 

exceeding limit dose levels. No developmental or reproductive effects were seen in the available 

studies at doses at or above limit dose levels. Based on this information, there is no concern, at 

this time, for increased sensitivity to infants and children to FD&C Red No. 40 when used as a

component of food contact sanitizing solutions applied to all food contact surfaces and a safety 
18

factor analysis has not been used to assess risk. For the same reason, EPA has determined that an 

additional safety factor is not needed. The Agency believes that any exposure to FD&C Red No. 

40 used as an inert ingredient in pesticide formulations would occur at a level much lower then 

an effect level noted in animal studies. 

EPA has determined that the available data supports an exemption from the requirement 

of a tolerance for residues of FD&C Red No. 40 (CAS Reg. No 25956-17-6) as a pesticide inert 

ingredient (dye) in antimicrobial formulations in accordance with 40 CFR 180.940 (a) as 

requested by the petitioner. There is a reasonable certainty that no harm to any population 

subgroup will result from FD&C Red No. 40 when used as inert ingredient in pesticide 

antimicrobial formulations applied to food contact surfaces in public eating places, dairy 

processing equipment, food processing equipment and utensils, as result of aggregate exposure. 

Therefore, it is recommended that a tolerance exemption for FD&C Red No. 40 (CAS Reg. No. 

25956-17-6) be established for use as an inert ingredient (dye), limited to 20 ppm, in 

antimicrobial formulations under 40 CFR 180.940(a), which includes use on food contact 

surfaces in public eating places, dairy processing equipment and food processing equipment and 

utensils. 

References

ChemIDplus, 2004, ChemIDplus Advanced. U.S. National Library of Medicine. National 

Institutes of Health. Department of Health and Human Services. Online Search Database 

http://chem.sis.nlm.nih.gov/chemidplus/

Search terms: FD&C Red No. 40. (May 7, 2014)

ECOSAR. 2000. Ecological Structure Activity Relationships Version 0.99g. Environmental 

Protection Agency. 

ECOTOX. U.S. Environmental Protection Agency. 2002. ECOTOX User Guide: 

ECOTOXicology Database System. Version 4.0 http://www.epa.gov/ecotox/

Search terms: CAS Reg Nos: 25956-17-6 (July 9, 2013)

EFSA. 1980. Joint Food and Agriculture Organization (FAO) and World Health Organization 

(WHO) Committee on Food Additives. “JECFA Evaluation – Allura RED AC”. MRID 

48779805

EFSA. 1981. Joint Food and Agriculture Organization (FAO) and World Health Organization 

(WHO) Committee on Food Additives. “Evaluation of certain food additives”. MRID 48779804

EFSA. 2009. Scientific Opinion on the re-evaluation of Allura Red AC (E 129) as a food 

additive. MRID 48779803
19

EPA, ISK Biosciences Corporation (2011); Information Submitted to Support Petition Number 

1E7843, Petition Proposing an Exemption from The Requirement of a Tolerance for Residues of 

FD&C Red No. 40 In or On Raw Agricultural Products and Food Products

EPI Suite. 2004. Estimation Programs Interface Suite Version 3.12 (August 17, 2004).

Environmental Protection Agency. http://www.epa.gov/opptintr/exposure/docs/episuite.htm

JECFA. 1980. Joint FAO/WHO Expert Committee on Food Additives. Allura Red AC. WHO 

Food Additive Series Vol. 15. http://www.inchem.org/documents/jecfa/jecmono/v15je02..htm

US Environmental Protection Agency (EPA), December 21, 2004, Action Memorandum, Dan Rosenblatt, Chief Minor 

Use, Inerts and Emergency Response Branch to Lois Rossi, Director Registration Division, Inert Ingredient Tolerance 

Reassessment – FD&C Blue No. 1, FD&C Red No. 40 and FD&C Yellow No. 5 (Tartrazine). MRID 48779801