July 24, 2014 UNITED STATES ENVIRONMENTAL PROTECTION AGENCY WASHINGTON, D.C. 20460

OFFICE OF CHEMICAL SAFETY AND POLLUTION PREVENTION



### MEMORANDUM SUBJECT: PC Code: 879709; Sweet Orange Peel Tincture; Human Health Risk Assessment and Ecological Effects Assessment to Support Proposed Exemption from the Requirement of a Tolerance When Used as Inert Ingredients in Pesticide Formulations.


### All PC Code : 879709 

### Decision No. : N/A DP Barcode: N/A Regulatory Action: Inert Tolerance Exemption; 40 CFR 180.910. 

### Petition No: IN

- 10547 All CAS Reg. No: 8028- 48-6 Inert Tracking No: N/A

FROM: Lisa Austin, Ph.D., Toxicologist ) Inert Ingredient Assessment Branch (IIAB)**~.ra--_,_** 

## __ _ Registration Division (7505P) 

### THROUGH: Kerry Leifer, Team Leader

# ~

# .rt--

Inert Ingredient Assessment Branch (IIAB) 

# h L Registration Division (7505P)
. 

# L TO: PV Shah, Ph.D., Branch Chief Inert Ingredient Assessment Branch (IIAB) Registration Division (7505P)EXECUTIVE SUMMARY EPA received a petition (PP#IN-10547) from AG-Chem Consulting (12208 Quinque Lane , Clifton VA 21 024), on behalf of Oro-Agri, Inc.(990 Trophy Club Drive, Trophy Club TX 76262) requesting an exemption from the requirement of a tolerance for sweet orange peel tincture (CAS Reg. No. 8028-48-6) for use as a s urfactant, fragrance and adjuvant inert ingredient up to 10% concentration in pesticide products on all pre- and post-harvest food commodities under 40 CFR part 180 .910. EPA published the notice of filing (NOF) for this petition in the Federal Register on July 19, 2013 (78 FR 43117). No comments were received in response to this notice. Sweet orange peel tincture and sweet orange peel oil are**chemi~ally** the same (EINECS No. 232-43 3 -8; CAS 8028-48-6). The only difference is the method of extraction. Sweet orange peel tincture is extracted in alcohol and the extraction process leads to the formation of a "tincture"
. Sweet orange peel oil is extracted by cold press expression. Both are extracted from the same plant, Citrus sinensis family Ru taceae. Both forms of the sweet orange peel extract contain d-limonene as its primary component(> 90 %) and lesser amounts ofmyrcene. The EPA has conducted a "Screening-Level Hazard Assessment" of a class of compounds called monoterpene hydrocarbons, d-limonene and myrcene are among the monoterpene hyrdrocarbons assessed. The chemical class was "based on structural similarity, similar molecular weights and functional groups and the expectation that inherent physicochemical, environmental and toxicological properties are predicted to be similar". That analysis also included a review of data on sweet orange peel oil since it is a complex mixture containing greater than 90% monoterpene hydrocarbons (including d-limonene and myrcene), and is expected to have "physicochemical , environmental and toxicological properties similar to the major components .. .limonene and myrcene". Therefore, the Agency assessed the potential toxicity of sweet orange peel tincture based on the available toxicity data for sweet orange peel oil, and where data for sweet orange peel oil is missing, relied upon available data for the monoterpene hydrocarbons chemical class. The acute oral and dermal toxicity of sweet orange peel. oil is low. The oral LD50 was >5,000 mg/kg in rats and rabbits, re spectively. In a 28 -day study with sweet orange peel oil, lesions in the non-glandular stomach and clinical chemistry were observed at 1 ,500 mg!kg/day (above the limit dose of 1 ,000 mg/kg/day) in rats. Lesions in the non-glandular stomach is attributed to the irritating property of the chemical. The NOAEL was 600 mg/kg/day. In a combined reproductive/developmental toxicity screening test with sweet orange peel oil, stillbirths and pup mortality were observed at 1,500 mg!kg/day. The offspring NOAEL was 750 mg/kg/day. Signs of toxicity were not observed in maternal rats, the NOAEL was 1,500 mg/kg/day.

2Evidence of mutagenicity was not observed in the Ames test. Although, evidence of mutagenicity was observed in mouse lymphoma assay, it was noted that cytotoxic concentrations were not reported and this could contribute to an inflated mutation rate. In addition, d-limonene was not mutagenic in an Ames test, mouse lymphoma, sister chromatid exchange nor in chromosome aberrations in Chinese hamster ovary cells assays. Therefore , based on the weight of evidence sweet orange peel tincture is not expected to be mutagenic. Evidence of immunotoxicity was not observed. Sweet orange peel oil exhibited no effects on cell-mediated or humoral immune response at doses up to 2 , 500 mglkg/day in a plaque-forming cell assay in mice. Although a carcinogenicity study was not available for sweet oi:ange peel tincture, carcinogenicity studies were available in the rat and mouse for d-limonene. An increased incidence oftumor formation was not observed in a 2 year carcinogenicity study on female rats or in male and female mice treated with d-limonene up to 1 ,000 mg/kg/day. An increased incidence oftubula r cell hyperplasia, adenomas, and adenocarcinomas ofthe kidney was observed in the male rat. However, these lesions are related to the accumulation ofthe alpha 2uglobulin protein which is specific to the male rat and is not relevant for human risk assessment. EPA has determined that reliable data show the safety of infants and children would be adequately protected if the FQPA SF were reduced to IX. Available toxicity data on sweet orange peel oil demons trated effects at 1,500 mg/kg/day and no toxicity at doses up to 600 mg/kg/day. Therefore , since there are no endpoints of concern for sweet orange peel tincture a qualitative dietary exposure risk assessment is appropriate. Sweet orange peel tinctu re is used as an inert ingredient in pesticide products that could result in residential exposure. However., based on the lack oftoxicity, a quantitative exposure assess ment from "residential exposures" was not performed. The Agency believes that amending the tolerance exemption to include 40 CFR 

### § 180.910 will not significantly/measurably increase occupational exposure . The change in use pattern would be expanded to include use as a surfactant, fragrance and adjuvant inert ingredient up to 10% concentration in pesticide products on all pre- and**post-harve~t** foo d commodit ies. Based on the absence of toxicity of sweet orange peel tincture , it is not necessary to quantitatively assess occupational exposure. Ecotoxicity and fate and effects data were not available for sweet orange peel tincture. However, limited data is available for d-limonene. The acute toxicity of d

-limonene ranges from slight to high for aquatic organisms
. It caused moderate acute toxicity in insects and mites
. It was biodegradable. Potential areas of environmental justice concerns, to the extent pos sible, were considered in this human health risk assessment, in accordance with U.S. Executive Order 12898, "Federal Actions to Address Environmenta l Ju stice in Minority Populations and Low-Income Populations," http: //www.eh.doe.gov/oepa/guidance / justice /eo12898
.pdf.
3Taking into consideration all available information on sweet orange peel tincture (CAS Reg. No. 8028-48

- 6), EPA concludes that there is a reasonable certainty that no harm will re sult to the general population or to infants and children from aggregate exposure to sweet orange peel tincture. Therefore, the establishment of an . exemption from tolerance under 40 CFR 180.91 0 for residues of sweet orange peel tincture for use as a surfactant, fragrance and adjuvant inert in gredient up to 1 0% concentration when used as an inert ingredient in pesticide formulations in pre- and post-harvest food commodities in the end-product formula tion can be considered safe under section 408 of the FFDCA. 

### I. BACKGROUND On Februar y 13, 2013 AG

-Chem Consulting (12208 Quinque Lane, Clifton VA 2 1 024), on behalf of Oro-Agri, Inc. (990 Trophy Club Drive, Trophy Club TX. 76262), submitted a petition (IN

- 1 0547) requesting an exemption from tolerance for sweet orange peel tincture (CAS Reg
. Nos. 8028-48-6) when used as a surfactant, fragrance and adjuvant inert ingredient up to 10% concentration in pesticide products on all pre- and post
-harvest food commodities under 40 CFR part 180.910. EPA published the Notice of Filing for this petition in the Federal Register on July 19 , 2013 (78 FR 43117)
. No comments were received in response to thi s notice. Currently, d

-limonene is exempted from the requirement of a tolerance when used in accordance with good agricultural practice as a solvent or fragrance (or occasionally active as an insecticide, insect repellent, and animal (dog and cat repellent)) ingredients in pesticide formulations under 4 0 CFR 180.910 and 180.930. It is also approved under 40 CFR 1 80.539 for used in insect-r epellent tablecloths and in insect-repellent strips u sed in food- or feed-handling establi shments. It i s a l so considered by FDA to be GRAS as a food additive when used as a synthetic flavoring substance and adjuvant (21 CFR 182.60). It is not registered for food or feed crop uses as an active ingredient, but can be used on compost and manure . This document provides an assessment ofthe risk to human health and the environment for for sweet orange peel tincture when used as an inert ingredient in pesticide formulations applied to pre- and post-harvest agricultural commodities. 

### II
. PHYSICAL AND CHEMICAL PROPERTIES Sweet orange peel tincture (CAS # 8028-48-6) is comprised ofterpenes, aldehydes and alcohols all present in concentrations of <2%
. The constituents of sweet orange peel tincture are found in Table 1(V ora et.al., 1983). 

### Table 1. 

### Sweet Orange Peel Tincture (CAS # 

### 8028-48-6) Constituents CAS# Constituents Typical Concentrat ion (% w/w) 5989-27-5 d-limonene >95% 123-35-3 myrcene <2%
480-56-8 a-pinene <1% 3387-41-5 Sabinene < 1% 99-83-2 Alpha- , Beta

- 

### < 1% 555-10-2 phell andrene(iso mers) 111-87-5 1-0ctanol 

### < 1% 124- 19-6 Nonanal 

### < 1% 78-70-6 Linalool 

### < 1% 106-23 -0 Citro nella! <1% 98-55-5 a-Terpineol <1% 112-3 1-2 Decanal <1% 106

-26-3 Neral <1% 14 1

-27

- 5 Geranial <1% 2111-75-3 Perillaldehyde <1% 1 12-14- 1 O ctyl acetate <1% 112-44 -7 Undecanal 

### < 1% 112-54-9 Dodecanal 

### < 1% 13877-93 -5 P-Caryophyllene 

### < 1% 3856-2 5

- 5 P- Copaene < 1 % 18794-84-8 P- Farnesene <1% 46 30-07-3 Valencene <1% 3779-62-2 P-Sinensa l <1% 17909-77

-2 a - Sinensal <1% 4674-50-4 N ootkatone <1% Some
.ofthe physical and chemical characteristics of sweet orange peel oil (CAS# 8008

- 57-9) are found in Ta ble 2. Table 2 . Physical and Chemical Properties of Sweet Orange Oil Parameter Value Source

Structure Limonene: 91-94% Myrcene : 2
.0-2.1% Chemical Name Sweet Orange Oil CAS # 8008-57-9 Molecul ar Weigh t As cited in Hi gh Production Common Names Citrus sinensis oil

Physical State liquid Vo lume SCREENING-LEVEL HAZARD -97 to -74 ° C (measured values for CHARACTERIZATION Melting Point (0 C) limonene); < - l0°C ( measured value f or · Monoterpene Hydrocarbons myrcene) Boiling Point ( 0 C) 167-l 72 ° C (measured values for myrcene) Category 1.43 at 20 ° C (measured value for Vapor Pressure (Pa at 20°C) limonene) ; 2 .01 at 25 ° C (measured value for myrcene) Partition Coefficient (Kow) 4.57 (measured value for li monene) ; 4.47 (measured value for terpinolene and sweet
5orange oil); 4 .1 7 (measured value for myrcene) Water Solubility (mg/liter at 13.8 at25°C (measured value for limonene); 5.6 at 25°C (measured value for 25°C) myrcene) Henry's Law Constant (kPa 0.032 (measured value for limonene) ; m

3/mol at 25 °C) 0.064 (measured value for myrcene)
3 . Hazard Assessment Sweet orange pee l tincture and sweet orange peel oil are chemically the same (EINECS No. 232-433-8; CAS 8028-48-6). The only difference is the method of extraction. Sweet orange peel tincture is extracted in alcohol and the extraction process leads to the formation ofa "tincture". Sweet orange peel oil is extracted by cold press expression. Both are extracted from the same plant, Citrus sinensis family Rutaceae. Both forms of the sweet orange peel extract contain d-limonene as its primary component(> 90 %) and lesser amounts ofmyrcene. The EPA has conducted a "Screening-Level Hazard Assessment" of a class of compounds called monoterpene hydrocarbo ns , d-limonene and myrcene are among the monoterpene hyrdrocarbons assessed. The chemical class was "based on structural similarity, similar molecular weights and functional groups and the expectation that inherent physicochemical, environmental and toxicological properties are predicted to be similar". That analysis also included a review of data on sweet orange peel oil since it is a complex mixture containing greater than 90% monoterpene hydrocarbons (including d-limonene and myrcene), and is expected to have "physicochemical, environmental and toxicological properties similar to the major components .. .li monene and myrcene"
. Therefore, the Agency assessed the potential toxicity of sweet orange peel tincture based on the available toxicity data for sweet orange peel oil, and where data for sweet orange peel oil is missing, relied upon available data for the monoterpene hydrocarbons chemical class. The following summaries were reproduced from the "Screening

-Level Hazard Characteri zation; Monoterpene Hydrocarbons Category" (USEP A 2009).
Sweet Orange Oi l CCASRN 8008

- 57-9) Acute Toxicity Studies Table 3. Acute Toxicity Profile of Sweet Orange Peel Oil Toxicity Study Type Results Category Acute Oral

- rat LD50 > 5000 mg/kg bw IV Acute Dermal

- Rabbit LD50 > 5000 mg/kg bw IV Acute Oral Toxicity Wistar rats 

### (10 males) were administered orange peel oil, sweet via gavage at 

### 5000 mg/kg-bw and observed fo r 14 days
. No deaths were reported
. The LDso (male)> 5000 mg/kg-bw
. 6Acute Dermal Toxicity New Zealand White rabbits (1 0, sex not reported) were administered orange peel oil, sweet dermally at 5000 mg/kg-bw on to their clipped, abraded abdominal skin for 24 hours and observed for 7 days. There were no mortalities. The 

### LDso > 5000 mg/kg-bw. Repeated Dose Studies 28-Day Oral Toxicity Study-rat In a 28-day study, Sprague Dawley rats (10/sex/dose) were administered orange peel oq, sweet , in 1% methyl cellulose, via gavage at 0, 240, 600 or 1500 mg/kg-bw /day
. No treatmentrelated effects were reported on survival , body weights or food consumption. T reatment
-related decreases in glucose were observed in males at the high dose and in females at the mid- and high dose
. Increases (significance not reported) 

### in serum albumin and total serum protein were observed in all treated females and in the high dose males. Kidney and liv er weights were increased in t he treated male groups and in the high dose female groups. Treatment-related lesions were observed in the nonglandular stomach ofthe high dose males and females, and in the kidney ofall treated males , characteristic of hyaline droplet formation and a lpha2u-globulin accumulation. Nephropathy associated with alph a2u

-g lobulin formation in male rats is not considered relevant to humans (US EPA, 1991). The LOAE L was 1500 mg/kg -bw/day based on lesions in the stomach and clinical chemistry. The NO AEL was 600 mglkg

-bw/day. Reproductive/Developmental Toxicity

-rat In a combined reproductive/developmental toxicity screening test j female Sprague

-Dawley rats . (10 /dose) were admini stered orange peel oil , sweet via gavage at 0, 375,750 or 1500 mg/kg/day for 7 days prior to and through mating, gestation, delivery and four days of lactation. Treated animals had decreased weight gains , and significantly decreased absolute and relative food consumption at 750 and 1500 mg/kg/day during the seven day pre-mating period. No treatment related effects on mating performance or fertility were observed at any dose level. A significant number of stillbirt hs and pup deaths were observed at 1500 mg/kg/day. No other treatment
-rela ted effects on offspring were observed. The maternal NOAEL was 1500 mglkg/day, the highest dose tested. The offspring LOAEL was 1 500 mg/kg/day based on stillbirths and pup mortality. The offspring NOAEL 750 mglkg/day. Mutagenicity In a reverse -mutation assay , Salmonella typhimurium strains TA98, TA 100, TA1535, TA1537 or TA1538 were exposed to orange peel oil, sweet at 5000 f.!g/plate in the presence and absence of metabolic activation. No information on the use of positive controls and cytotoxic concentrations was provided. No evidence ofmutagenic activity was reported at any concentration tested, with or without metabolic activation. In a forward mutation assay, mouse lymphoma L5178Y TK+/

- cells were exposed to orange peel oil at concentrations ranging from 40 to 200 )lg/mL in the presence and absence ofmetabolic

7activation. Increase s in mutation were observed with and without metabol ic activation. Cytotoxicity (co ncentration not sta ted) was also observed. Thi s could contribute to an inflation of mutatio n s. Immunotoxicity In a plaque

-formin g cell (PFC) assay, female B6C3 Fl mice were admini stered orange peel oil via at gav age 0, 625 , 1250 or 2500 mg/kg bw daily for 5 days to determine effects o n humoral and cellmediated immune responses. A host re s istance assay (Li steria monocytogenes bacterial challenge) was used to assess cell-mediated immunity w hile the antibody plaque f o rming cell response to sheep erythrocytes was used to measure humoral immunity. B ody weights, lympho id organ weights and spleen cellularity were also evaluated
. In the absence of modulation of the PFC response , these effects were not considered as indicators of immunotoxicity. Orange oil had no e ffects on cellmediated or humoral immune response at any do se level te sted. The NOAEL 

## = 2500 mg/kg/day, the highest dose tested).

Table 4. Toxicology Profile for Sweet Orange Oil Guideline No./ Study MRID No. (year)/ Results Type Classification /Doses 28-week-Oral Toxicity 0, 240 , 600 or 1500 NOAEL: 600 mglkglday Study

- Rat-gavage mg/kglday LOAEL : 1500 mglkglday based on lesions in the stomach and clinical chemistry. Developmental Toxicity

- 0 , 250 , 500 and 1000 Matern al NOAEL 

### = 500 mglkglday. Sprague- Daw ley rat
- mglkg/day gavage Maternal LOAEL 

### = I 000 mglkg/day based on reduced body weight , body weight gain , food consumption. Developmental NOAEL' 

### = I 000 mglkg/day. Developmental LOAEL was not estab lished
. Re production/Development 0 , 375, 750 or 1500 Mate rnal NOAEL 

### = 1500 mglkg/day. a ! Toxicity -Rat -gavage mglkglday Maternal LOAEL was not established. Developmental NO AEL = 750 mglkg/day. Developmental LOAEL = 1500 mg/kg/ day based on on stillbirths and pup

**ort~lity.** Ames Test 0 and 5000 ug/plate Negative.

Mouse Lymphoma Assay 40-200 ug/ml Increases in mutation were observed w ith and without metabolic activation. Cytotoxicity (concentration not stated ) was also observed.

8

### Table 4. Toxicology Profile for Sweet Orange Oil Guideline No./ Study MRID No. (year)/ Results Type Classification /Doses Plaque-Forming Cell Assay 0 , 625 , 1250 or 2500 mglkg NOAEL: 2500 mg/kglday -Mice bw LOAEL was not established.



### D. Toxicity Endpoint Selection The avai lable toxicity studies indicate that sweet orange peel tincture has low toxicity. These data demonstrated effects at 1500 mg/kg/day (above the limit dose) and no toxicity at doses up to 600 mg/kg/day. Therefore, since no endpoint of concern was identified for sweet orange peel tincture, a qualitative risk assessment is appropriate. 

### E. Special Considerations for Infants and Children Section 408 ofthe FFDCA provides that EPA shall apply an additional margin of safety for infants and children in the case ofthreshold effects to account for prenatal and post natal toxicity and the completeness of the database on toxicity and exposure unless EPA determines that a different margin of safety will be safe for infants and children
. . EPA concludes that the FQP A safety factor could be reduced to 1X for sweet orange peel tincture for the following reasons:
1. The database is considered adequate for FQPA assessment based on the toxicity information on sweet orange oil and d-limonene. The following accepta ble studies are available: Subchronic (rat) Combined Reproductive/Developmental toxicity study (rat) Immunotoxicity s tudy (rat)

u . Quantitative fetal susceptibility was observed in combined repro duction/developmental study with rats administered sweet orange peel oil. In rats , maternal toxicity was not observed at 1500 mg/kg/day (above 1000 mg!kg/day, the limit dose). Developmental effects (stillborn pups and pup mortality) were observed at 1500 mg/kg/day. However, there is no concern for these effects since they occur above the limit do se.
111. Neurotoxicity studies were not available for review. However, signs of potential neurotoxicity were not observed in any of the s ubmitted studies with sweet orange oil or d-limonene. EPA concluded that neither a neurotoxicity nor a deve lopmental neurotoxicity study is required at this time.

9iv. An immunotoxicity study was available for review. Evidence of immunotoxicity was not observed in a plaque-forming cell assay in mice treated with sweet orange oil. Therefore, EPA concluded that the immunotoxicity study is not required at this time. Taking into consideration the available information , EPA concludes the additional 1OX FQPA safety factor can be reduced to IX.
4. Exposure Assessment In evaluating dietary exposure to sweet orange peel tincture, EPA considered exposure under the petitioned for exemptions from the requirement of a tolerance. EPA assessed dietary exposures from sweet orange oil in food as follows: Dietary Exposure Dietary exposure can occur from eating foods containing residues of sweet orange peel tincture. Because no hazard endpoint of concern was identified for the acute and chronic dietary assessment (food and drinking water), a quantitative dietary exposure risk assessment was not conducted. Residential Exposure (non-dietary exposure) The term " residential exposure" is used in this document to refer to non-occupational, nondietary exposure (e.g. , textiles (clothing and diapers), carpets , swimming pools , and hard surface disinfection on walls , floors, tables). Sweet orange peel tincture is used as an inert ingredient in pesticide products that could result in short- and intermediate-term residential exposure . However, based on the lack oftoxicity, a quantitative exposure assessment from residential exposures was not performed, 5. Aggregate Risk Based on the lack of any endpoints of concern, 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 sweet orange peel tincture residues. 6. Occupational Exposure/Risk Pathway The Agency believes that amending the tolerance exemption to include 40 CFR 

### § 180.910 w ill not significantly / measurably increase occupational exposure. The change in use pattern would be expanded to include use as a surfactant, fragrance and adjuvant inert ingredient up to 10% concentration in pesticide products on all pre- and post-harvest food commodities . B ased on the regulatory history of d-limonene, the major component of sweet orange peel tincture and no new toxicological information to indicate otherwise , it is not necessary to quantitatively assess occupational exposure.

107. Cumulative Exposure: Section 408(b)(2)(D)(v) ofFFDCA requires that, when considering whether to establish, modify, or revoke a tolerance, the EPA consider "available information" concerning the cumulative effects of a particular pesticide's residues and "other substances that have a common mechanism oftoxicity." Unlike other pesticides for which EPA has followed a cumulative risk approach based on a common mechanism of toxicity, EPA has not found sweet orange peel tincture to share a common mechanism oftoxicity with any other substances and sweet orange peel tincture does not appear to produce a toxic metabolite produced by o ther substances. The major component of sweet orange peel tincture is d-limonene. An endpoint of concern was not identified for d

- limonene in various dietary and non-dietary exposure scenarios. Additionally, the risk to aggregate exposure of d-limonene was below the Agency's concern. For information regarding EPA's efforts to determine which chemicals have a common mechanism oftoxicity and to evaluate the cumulative effects of such chemica ls, see the policy statements released by EPA's Office of Pesticide Programs concerning common mechanism determinations and procedures for cumulating effects from substances found to have a common mechanism on EPA's webs ite at http://www
.epa.gov/pesticides/cumulative/. 8. Environmental Fate The environmental fate summary was reproduced from International Programme on chemical Safety's Concise Internationa l Chemical Assessment Document No. 5 Limonene (1998). Limonene was completely biodegraded in 8 days in a laboratory study using unacclimated soil obtained from a coniferous forest. It was also shown to be readily biodegradable using the modified MITI test (OECD 301 C)(Limonene (CICADS)). Ecotoxicity Ecotoxicity summary was reproduced from International Programme on chemical Safety's Concise International Chemical Assessment Document No. 5 Limonene ( 1998). Aquatic envirorunent The acute toxicity of d-limonene ranges from s light to high for aquatic organisms. The lowest acute toxicity values (EC50 or LC50) identified were approximately 0.4 mg/litre for 

### Daphnia (US EPA, 1990b) and 0. 7 mg/litre for fish. The no -observed-effect concentration (NOEC) for green algae is approximately 4 mg/litre. The acute toxicity (EC50 or LC50) of dipentene to 

### Daphnia and fish is about 50-70 time s lower than that for d-limonene. No studies were identified on the chronic toxicity of limonene to aquatic organisms. Terrestrial environment Limonene generally has moderate acute toxicity in insects and mites. The acute toxicity of dlimonene to earthworms 

### (Eiseniafoetida Savigny) was high (LC50 

### = 6.0 ppm; mg/kg)
.

. 11Sublethal effects (i.e. abnormal rebounding of medial giant fibre pathway [MGF] impulses and spontaneous lateral giant fibre pathway [LGF] spiking) were observed following exposure of earthworms to 4.2 ppm (mg/kg) limonene. Limonene has lo w subacute toxicity to bobwhite quail 

### (Colinus virginianus) exposed via the diet (LC50 

### > 5620 ppm; mg/kg). 9 . Analytical Methods Analytical methods are not available for residue measurement. 10. Environmental 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 Low- Income Populations," http: //www.eh.doe.gov/oepalguidance /justice/eol2898.pdf). As a part of every pesticide risk assessment, OPP considers a large variety of consumer subgroups according to well -e stablished procedures. In line with OPP policy, HED estimates risks to population subgroups from pesticide exposures that are based on patterns ofthat subgroup's food and water consumption, and activities in and around the home that involve pesticide u se in a res i dential 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. Thes e data are analyzed and categorized by subgroups based on age, season ofthe year, ethnic group, and region ofthe 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
. 11. Human Studies This assessment relies in part on data from studies in which adult human subjects were intentionally exposed to a pesticide. These studies, listed below, have received the appropriate ethical review for use in risk assessment. The PHED Task F orce, 1998. The Pesticide Handler E xposure Database (PHED), Version 1.1. Task Force members: Health Canada, U.S. Environmental Protection Agency, the California Department of Pesticide regulation, and the American Crop Protection Association; released August 1998
. 12. Risk Characterization

12Sweet orange peel tincture and sweet orange peel oil are chemically the same (EINECS No. 232-433-8; CAS 8028-48-6). The only difference is the method of extraction. Sweet orange peel tincture is extracted in alcohol and the extraction process leads to the formation of a "tincture". Sweet orange peel oil is extracted by cold press expression. Both are extracted from the same plant, Citrus sinensis family Rutaceae. Both forms ofthe sweet orange peel extract contain d-limonene as its primary component(> 90 %) and lesser amounts ofmyrcene. The EPA has conducted a "Screening-Level Hazard Assessment" of a class ofcompounds called monoterpene hydrocarbons, d-limonene and myrcene are among the monoterpene
- hyrdrocarbons assessed. The chemical class was "based on structural similarity, s imilar molecular weights and functional groups and the expectation that inherent physicochemical, environmental and toxicological propert ies are predicted to be similar". That analysis also included a review of data on sweet orange peel oil since it is a complex mixture containing greater than 90% monoterpene hydrocarbons (including d-limonene and myrcene), and is expected to have "physicochemical, environmental and toxicological properties similar to the major components .. .limonene and myrcene". Therefore , the Agency assessed the potential toxicity of sweet orange peel tincture based on the available toxicity data for sweet orange peel oil, and where data for sweet orange peel oil is missing, relied upon available data for the monoterpene hydrocarbons chemical class. The acute oral and dermal toxicity of sweet orange peel oil is low. The oral LD50 was >5,000 mg/kg in rats and rabbits, respectively. In a 28-day study with sweet orange peel oil, lesions in the non-glandular stomach and clinical chemistry were observed at 1,500 mg/kg/day (above the limit dose of 1 ,000 mg/kg/day) in rats. Lesions in the non-glandular stomach is attributed to the irritating property of the chemical. The NOAEL was 600 mg/kg/day. In a combined reproductive/developmental toxicity screening test with sweet orange peel oil, stillbirths and pup mortality were observed at 1,500 mg/kg/day. The offspring NOAEL was 750 mg/kg/day. Signs of toxicity were not observed in maternal rats, the NOAEL was 1 ,500 mg/kg/day. Evidence of mutagenicity was not observed in the Ames test. Although, evidence of mutagenicity was observed in mouse lymphoma assay, it was noted that cytotoxic concentrations were not reported and this could contribute to an inflated mutation rate. In addition, d-limonene was not mutagenic in an Ames test, mouse lymphoma, sister chromatid exchange nor in chromosome aberrations in Chinese hamster ovary cells assays. Therefore, based on the weight of evidence sweet orange peel tincture is not expected to be mutagenic. Evidence of immunotoxicity was not observed. · Sweet orange peel oil exhibited no effects on cell-mediated or humoral immune response at doses up to 2,500 mg/kg/day in a plaque-forming cell assay in mice. Although a carcinogenicity study was not avai lable for sweet orange peel tincture, carcinogenicity st udies were available in the rat and mouse for d-limonene. An increased inc idence of tumor formation was not observed in a 2 year carcinogenicity study on female rats or in male and female mice treated with d-limonene up to 1 ,000 mg/kg/day. An increased
13incidence of tubular cell hyperplasia, adenomas, and adenocarcinomas ofthe kidney was observed in the male rat. However, these lesions are related to the accumulation of the alpha 2uglobulin protein which is specific to the male rat and is not relevant for human risk assessment. EPA has determined that reliable data show the safety of infants and children would be adequately protected ifthe FQPA SF were reduced to IX. Available toxicity d ata on sweet orange peel oil demonstrated effects at 1,500 mg/kg/day and no toxicity at doses up to 600 mglkg/day. Therefore , since there are no endpoints of concern for sweet orange peel tincture a qualitative dietary exposure risk assessment is appropriate. Sweet orange peel tincture is used as an inert ingredient in pesticide products that could result in residential exposure. However, based on the lack of toxicity, a quantitative exposure assessment from "res idential exposures" was not performed. The Agency believes that amending the tolerance exemption to· include 40 CFR 

### § 180.910 will not significantly /measurably increase occupational exposure. The change in use pattern would be expanded to include use as a surfactant, fragrance and adjuvant inert ingredient up to 10% concentration in pesticide products on all pre- and post-harvest food commodities. Based on the absence oftoxicity of sweet orange peel tincture, it is not necessary to quantitatively assess occupational exposure. Ecotoxicity and fate and effects data were not available for sweet orange peel tincture. However, limited data is available for d

-limonene. The acute toxicity of d-limonene ranges from slight to high for aquatic organisms. I t caused moderate acute toxicity in insects and mites. It was biodegradable. 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 /eol2898.pdf. Taking into consideration all available information on sweet orange peel tincture (CAS Reg. No. 8028-48-6) , EPA concludes that there is a reasonable certainty that no harm will result to th e general population or to infants and children from aggregate exposure to sweet orange peel tinctur e. Therefore, the establishment of an exemption from tolerance under 40 CFR 180.910 for residues of sweet orange peel tincture for use as a surfactant, fragrance and adjuvant inert ingredient up to 10% concentration when used as an inert ingredient in pesticide formulations in pre- and post-harvest food commodities in the end-product formulation can be considered safe under section 408 ofthe FFDCA.

14REFERENCES International Programme On Chemical Safety. Concise International Chemical Assessment Document No.5 Limonene. 1998. USEPA (1994) Reregistration Eligibility Decision (RED): Limonene. USEPA (2003) d-Limonene. Exposure and Risk Assessment on Lower R isk Pesticide Chemicals. d-Limonene. USEPA (2009) Screening-Level Hazard Characterization; Monoterpene Hydrocarbons Category
. USEPA (2010) d-Limonene. Human-Health Assessment Scoping Document in Support of Registration Review. DP Barcode D374492. USEPA EPA 738-R-94-034. 1994 Reregistration Eligibility Decision (RED): Limonene. Vora, J.D. , Matthews, R.F
. , Crandall, P.G
. , Cook, R. (1983). Preparation and Chemical Composition of Orange Oil Concentrates. Journal of Food Science, (48) : 14-16. Friedrich K, Delgado IF , Santos LM , Paumgartten F J. Assessment of sensitization potential of monoterpenes using the rat popliteal lymph node assay . Food Chern Toxicol. 2007 Aug;45 (8):1516-22. Epub 2007 Feb 17. Hoberman, A. (1992) Developmental Toxicit y (Embryo-Fetal Toxicity and Teratogenic Potential) Study ofD-Limonene Technical Administrered Orally Via Gavage to Crl:CD®BR VAF/Plus® Presumed Pregnant Rats . Argus Research Laboratorie s, Inc., 905 Sheehy Drive Horsham, PA 19044. Laboratory Project ID 2 119-001. MRID 42316301

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