       UNITED STATES ENVIRONMENTAL PROTECTION AGENCY
			WASHINGTON, D.C.  20460




						                   OFFICE OF CHEMICAL SAFETY 							                                                            AND POLLUTION PREVENTION						




MEMORANDUM  


DATE:		December 10, 2018



SUBJECT:      IN-11074; Waxes and Waxy Substances, Rice Bran, Oxidized: Human
            Health Risk and Ecological Effects Assessment of a Food Use Pesticide Inert Ingredient 
             
		CAS Reg. No. 1883583-80-9; 	 
      PC Code:  800245;     
      Decision 533871


FROM:	Deirdre Sunderland, MHS, Industrial Hygienist
		Chemistry, Inerts & Toxicology Assessment Branch (CITAB)
		Registration Division (RD); 


THROUGH:	Kerry B. Leifer, Team Leader
		CITAB/RD 	


TO:		Donna Davis, Acting Branch Chief
		CITAB/RD 










                               Table of Contents

Executive Summary.....................................................................................................3

 Background.........................................................................................................5
2.0 Inert Ingredient Profile..............................................................................................5
   2.1 Summary of Uses.................................................................................................5
   2.2 Physical and Chemical Properties................................................................................5
   2.3 Metabolism and Pharmacokinetics.............................................................................6
   2.4 Surrogate Data....................................................................................................7

3.0 Hazard Assessment.................................................................................................9
   3.1 Acute Toxicity.....................................................................................................9
   3.2 Subchronic Toxicity.............................................................................................10
   3.3 Chronic............................................................................................................10
   3.4 Reproductive and Developmental Toxicity..................................................................13
   3.5 Neurotoxicity.....................................................................................................16
   3.6 Mutagenicity/Cytotoxicity......................................................................................16
   3.7 Carcinogenic Potential..........................................................................................17
   3.8 Endpoint Selection and Levels of Concern..................................................................17
   3.9 FQPA Safety Factor Considerations..........................................................................18

4.0 Exposure Assessment.............................................................................................18
   4.1 Dietary Exposure...............................................................................................18
   4.2 Cancer Exposure.................................................................................................18
   4.3 Residential Exposure...........................................................................................18
   4.4 Occupational Exposure........................................................................................19

5.0 Aggregate Risk....................................................................................................19

6.0 Cumulative Risk...................................................................................................19

7.0 Environmental Fate................................................................................................19

8.0 Ecological Effects..................................................................................................20
   8.1 Fish Toxicity......................................................................................................20
   8.2 Invertebrate Toxicity.............................................................................................20
   8.3 Algal Toxicity....................................................................................................20
   8.4 Terrestrial Toxicity...............................................................................................20

9.0 Risk Characterization..............................................................................................21





EXECUTIVE SUMMARY

On August 21, 2017, Spring Trading Company, on behalf of Clariant Corporation, submitted a petition to the Environmental Protection Agency, herein referred to as the agency or EPA, to amend 40 CFR 180.910, 180.930, and 180.940(a) by establishing an exemption from the requirement of a tolerance for the use of the inert ingredient, waxes and waxy substances, rice bran, oxidized, (CAS Reg. No. 1883583-80-9), herein referred to as rice bran wax oxidized or RBWO, in pesticide formulations applied pre-and post-harvest, to animals, and in antimicrobial formulations (food-contact surface sanitizing solutions). 

Rice bran wax (RBW) is a hard crystalline vegetable wax processed from rice bran oil obtained from rice husks. When oxidized, the polarity of RBW increases by partial cleavage of the esters and oxidation of the resulting alcohols leading to the formation of carboxylic acids. Rice bran wax, oxidized consists mainly of long chain fatty acids, alcohols, and esters. 

Rice bran wax, oxidized, is used in agricultural applications (e.g. seed coatings, fertilizer coatings, adjuvants), polish and care applications (e.g. shoe, floor, and car care), wax emulsions and dispersions, coating and inks, textiles, leather, and paper. It currently has no approved pesticidal uses. According to the submitter, rice bran wax, oxidized will be used as an inert ingredient in agricultural formulations as a flow aid, surface protection, film-forming, carrier, coating agent, and adjuvant. 
 
Available studies include an acute oral toxicity study, and dermal irritation, eye irritation, and dermal sensitization studies. Rice bran wax, oxidized was also tested in an Ames assay to determine the potential for reverse gene mutation. No subchronic or chronic studies are available for RBWO. Since data on rice bran wax, oxidized is limited, surrogate data on various other long chain fatty acids, long chain fatty alcohols and long chain esters were submitted to support the safety finding for rice bran wax, oxidized. Submitted data consisted of various subchronic, chronic, and reproductive/developmental studies on chemicals including carnauba wax, rice bran wax, D-002 (a mixture of primary long chain alcohols isolated from beeswax), D-003 (a mixture of long chain fatty acids isolated from sugar cane wax), docosanol, docosanoic acid, and policosanol.

The potential for absorption of rice bran wax via the gastrointestinal (GI) tract is limited. The long-chain fatty acids, alcohols, and esters present in plant-based waxes are generally thought to be poorly absorbed in the GI tract as uptake decreases as chain length and hydrophobicity increase. RBW is being used as a surrogate for RBWO based on its similar physical and chemical properties and its expected potential for toxicity; therefore, it is unlikely that RBWO would be systemically available. 

No endpoint of concern was identified for any of the acute studies conducted. In addition, no endpoint of concern was determined in any of the studies up to the limit dose of 1000 mg/kg/day. There was no evidence of carcinogenicity in any of the studies presented including chronic studies and studies on mutagenicity and cytotoxicity. In addition, no neuropathological changes or effects were reported in any of the studies. The agency does not believe RBWO will be carcinogenic or neurotoxic. 

Since no endpoint of concern was identified in acute, subchronic, or chronic studies and because rice bran wax, oxidized is not expected to be absorbed by the body, a quantitative risk assessment for RBWO was not performed. 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. 

While residential and occupational exposure are also possible, there are no toxicological effects of concern in available studies and therefore, it is not necessary to conduct a quantitative assessment of residential (non-occupational) exposures and risks. Similarly, a quantitative occupational risk assessment is not necessary. 

Based on ecological and fate data the agency concluded that rice bran wax, oxidized does not pose an ecological risk to terrestrial or aquatic species. Considering all evaluated toxicity studies coupled with the expected exposure from the use of this chemical as inert ingredient in pesticide products, EPA concludes that there is a reasonable certainty that no harm will result to the general population, including infants and children, from aggregate exposure to residues of waxes and waxy substances, rice bran, oxidized when used as an inert ingredient in pesticide formulations under 40 CFR 180.910, 180.930, and 180.940(a).
 


































 1.0       BACKGROUND

On August 21, 2017, Spring Trading Company, on behalf of Clariant Corporation, submitted a petition to the EPA, to amend 40 CFR 180.910, 180.930, and 180.940(a) by establishing an exemption from the requirement of a tolerance for the use of the inert ingredient waxes and waxy substances, rice bran, oxidized, (CAS Reg. No. 1883583-80-9)  herein referred to as rice bran wax oxidized or RBWO for use in agricultural, animal, and antimicrobial pesticide formulations as a flow aid, surface protectant, film-former, carrier, coating agent, or adjuvant.

2.0       INERT INGREDIENT PROFILE

2.1    Summary of Uses

According to the submitter, rice bran wax oxidized is currently used in "agricultural applications (e.g. seed coatings, fertilizer coatings, adjuvants), polish & care applications (e.g. shoe, floor and car care), wax emulsions & dispersions, coating & inks, textile, leather, paper." Rice bran wax oxidized has no currently approved pesticidal uses. According to the U.S. National Institutes of Health's Household Product Database (https://householdproducts.nlm.nih.gov), rice bran wax oxidized is not found in any household consumer goods. 
 2.2    Physical and Chemical Properties

Some of the physical and chemical characteristics of rice bran wax oxidized, are found in Table 1 below.  When oxidized, crude rice bran wax is changed in color from brown to white. The brown color is due to a resinous material in the crude rice bran wax. Oxidation increases the polarity of the product by partial cleavage of the esters and oxidation of the resulting alcohols leading to the formation of carboxylic acids. For comparison, the physical and chemical properties for rice bran wax have also been included in Table 1. 

The composition of the two oxidized rice bran waxes were tested and the results showed one consisted primarily of rice bran wax carboxylic acids with chain lengths of >19 (31.68%) and rice bran wax esters (39.77%).  The other also consists primarily of rice bran wax carboxylic acids with chain lengths of >19 (19.83%) and rice bran wax esters (72.95%) but in different quantities. The alcohols making up the wax esters in both products are saturated with chain lengths of C-24 to C-36 with C-30 (triacontanol) having the highest content. The long chain fatty acids are primarily C-24 with lesser amounts of C-22, 26, 28, 30, 32 and 34 for both test substances.

The chemical constituents of RBW are mainly saturated monoesters (C-46 to C-60) of long chain fatty acids (C-22 to C-26) and long chain fatty alcohols (C-26 to C-30). The three primary esters in rice bran wax are mericyl cerotate (43 to 45%), ceryl cerotate (21 to 22%) and isoceryl isocerotate (9-10%). (Maru, Surawase, & Bodhe, 2012) The alcohols forming these esters, mericyl alcohol or triacontanol, ceryl alcohol or hexacosanol and isoceryl alcohol are C-30, C-26 and C-27 alcohols, respectively.  Cerotic acid or hexacosanoic acid has a fatty acid tail of 24 carbons.  

Table 1.  Physical and Chemical Properties of Rice Bran Wax, Oxidized and Rice Bran Wax 
Parameter
RBWO
RBW
CAS Reg. No. 
1883583-80-9
8016-60-2
Physical State
solid
solid
Melting Point
74.6-78.6°C
75-85.5
Density 
0.99-1.01 @23°C
0.96 @20°C
Boiling Point
273-366 °C Thermal decomposition
343°C
Vapor Pressure 
1 x10[-5] mm Hg (@25°C)
N/A
Water Solubility
<1 g/L
Insoluble
 2.3    Metabolism and Pharmacokinetics 
No information was submitted on the absorption, distribution, metabolism, or excretion of the oxidized form of rice bran wax; however, the following was noted in GRAS Notice (GRN) #655 (https://www.fda.gov/downloads/Food/IngredientsPackagingLabeling/GRAS/NoticeInventory/ucm516030.pdf) for rice bran wax. 

      The potential for absorption via the gastrointestinal (GI) tract is limited for rice bran wax. The long-chain fatty acid esters present in plant-based waxes such as rice bran wax and carnauba wax are generally thought to be poorly absorbed in the GI tract (EFSA, 2012 a,b); uptake is thought to decrease as chain length and hydrophobicity increase (Hargrove et al., 2004). While some species have adapted to the use of wax esters as energy sources, humans are thought to be inefficient at this process (Hargrove et al., 2004). Supporting this conclusion, EFSA's Scientific Panel on Food Additives and Nutrient Sources (ANS) added to Food (EFSA, 2012b) reviewed one study evaluating the bioaccumulation of carnauba wax in rats as part of a 90-day toxicity feeding study; they note that the results of this study "indirectly suggested that lipid like components from the wax are not accumulated in tissues" (Edwards, 1998 as cited in EFSA, 2012b). EFSA's Panel on Contaminants in the Food Chain (CONTAM; EFSA, 2012a) considered that absorption of carnauba wax into the GI tract is likely to be low or will not occur, so that metabolism of wax by digestive enzymes or intestinal microbiota is unlikely. In situations where minor amounts of digestion occur, it has been shown that the resulting free fatty acid and alcohol can be absorbed by the epithelium and incorporated into normal cellular metabolic pathways (EFSA, 2012 a,b; Hargrove et al., 2004). If a small amount of rice bran wax was absorbed and metabolized to some degree into ethyl alcohol (ethanol), exposure to ethanol would be low in contrast to exposure from the daily diet. Consumers are routinely exposed to incidental amounts of ethanol from consumption of food items such as orange juice, soft drinks, and breads. GRN 151 (FDA, 2004) received a "no questions letter" for the use of ethyl alcohol as a preservative in the filling used in shelf-stable croissants at a concentration of 3,000 ppm. In addition, GRN 151 reported ethanol levels in ripening fruit and fruit juice ranging from 117 to 1,900 ppm and Logan and Distefano (1998) reported levels of ethanol in various baked good ranging from 0 -1.66 %. It is reasonable to conclude that any absorption of rice bran wax via the oral route of exposure would be negligible and does not present any safety concern.
For the purpose of making a safety finding on RBWO, data on RBW is being used as a bridge. Based on its similar physical and chemical properties and similar composition the two substances would be expected to behave similarly in the body. RBW is chemically oxidized to form RBWO. The small portion of RBW that is absorbed is expected to undergo this same process by endogenous enzymes like esterases and oxidases. Subsequently, RBWO is expected to be poorly absorbed via the GI tract as is RBW. 
2.4    Surrogate Data
Limited toxicity data are available on rice bran wax oxidized. Therefore, the petitioner has submitted information on various surrogate chemicals in lieu of studies on rice bran wax oxidized. RBW is chemically oxidized to form RBWO. This same process is believed to occur naturally in mammalian organisms by endogenous enzymes like esterases and oxidases.
Rice bran wax (RBW) is a hard, crystalline vegetable wax obtained from rice husks. It is processed from the rice bran oil that comes from the husks. The exact composition of RBW is not fully established as the results on studies looking at the compositional nature of RBW have been varied. According to Vali (2005), 

      Iwama and Maruta (6) fractionated RBW into hard wax (38.5%) and soft wax (11.2%) and found the composition of hard wax to be a mixture of esters of C22, C24, and C34 FA and C18 to C34 fatty alcohols (FAL). Yoon and Rhee (7) reported the presence of a substantial amount of dark brown resin-like matter in both soft and hard wax and only C22 to C30 FAL and C16 to C26 FA. Belavadi and Bhowmick (8) concluded that some esters exist in polymeric form involving aromatic moieties. Ito et al. (9) reported that RBW contains esters of branched-chain aliphatic C32, C34, and C36 FAL. Wang et al. (10) reported that refined RBW contains C22 - C38 FA and C14 - C38 FAL. The inconsistency of the reports on the composition of RBW is likely due to the lack of an efficient purification method and a suitable analytical technique."

The Vali article also states:

      "The resinous matter was a mixture of aliphatic aldehydes, fatty alcohols, and FA. High-temperature GC analysis of the purified rice bran wax indicated that it contained 11 major and 9 minor types of saturated wax esters. The major and minor peaks contained C44 - C64 and C45 - C59 wax esters, respectively. Rice bran wax was mainly a mixture of saturated esters of C22 and C24 FA and C24 to C40 aliphatic alcohols, with C24 and C30 being the predominant FA and fatty alcohol, respectively. The alcohol portion of the wax esters also contained small amounts of branched and odd carbon number fatty alcohols."

In addition to RBW, studies on other chemicals with similar compositions have been submitted to support the safety assessment of RBWO. The names of these chemicals along with a brief description are listed in Table 2 below. Previous work has also been done to compare RBW to Carnauba Wax. A table (i.e., Table 3), from a 2012 article by Maru, Surawase, and Bodhe in the International Journal of Pharmaceutical and Phytopharmacological Research, making this comparison, is presented below.









                   Table 2: Surrogate Chemical Descriptions
Chemical
Description
Rice Bran Wax
1. Derived from rice (oryza sativa). Rice bran wax is hard non- tacky wax and is the byproduct of rice bran oil refinery. The oil is extracted from Rice bran which is the outer covering of the rice kernel after removing the husk. 
2. Contains mainly saturated monoesters (C-46 to C-60) of long chain fatty
acids (C-22 to C-26) and long chain fatty alcohols (C-26 to C-30). Main esters are myricyl cerotate (43-45 %), ceryl cerotate (21-22 %) and Isoceryl isocerotate (9-10 %).
Carnauba wax
1. Derived from the leaves of the palm Copernicia prunifera
2. Monoester fractions (54-84%) very similar to rice bran wax (87-98%) and RBWO (72.95%). 
3. Composition: Hydrocarbon 0.3-1%, aliphatic esters 38-40%, monohydric alcohols 10-12%, w-hydroxy aliphatic esters 12-14%, p-methoxycinnamic aliphatic diesters 5-7%, p-hydroxycinnamic aliphatic diesters 20-23%, an uncombined triterpene type diol 0.4% and uncombined acids and other unknown constituents 5-7%. 
D-002
1. Mixture of primary long chain alcohols isolated from beeswax.  
2. The composition varies slightly with batch but consists of approximately of 26.6% triacontanol, 17.5% octaconsanol, 17% C-32 primary alcohol (dotriacontanol), 15.3% hexaconsanol and 13.2% C-24 primary alcohol (tetracosanol).  
D-003
1. Mixture of long chain fatty acids isolated from sugar cane wax. 
2. Composed of 1-octacosanoic acid (40.0%; C-28), 1-triacontanoic acid (18 %; C-30), 1-nonacosanoic acid (3%; C-29), 1-tetratricontanoic acid (12%; C-34) and other long chain fatty acids in lesser amounts
Docosanoic acid 
(behenic acid),
1. A straight chain fatty acid with a chain length of 22 
2. It is a constituent of rice bran wax oxidized and is also found in beeswax and other parts of the human diet. 
3. Found naturally in humans (e.g, skin, earwax, and vernix caseosa- sebaceous secretion that covers the human fetus)
Policosanol
1. Mixture of higher primary aliphatic alcohols                                                                 2. Isolated from sugar cane, also found in beeswax. 
3. Consists of 66% octaconsanol, 12% triacontanol and 7% hexacosanol.   Octaconsanol, triacontanol and hexaconsanol are C-28, C-30 and C-26 long chain aliphatic alcohols, respectively
Docosanol 
(behenyl alcohol),
C-22 primary long chain fatty alcohol
Triacontanoic acid
C-30 primary long chain fatty acid
Triacontanol
C-30 primary long chain fatty alcohol
Mixtalol
A mixture of 7-10% tetracosanol (C-24), 12-16% hexacosanol (C-26), 15-20% octaconsanol (C28), 24-30% triacontanol (C30), 11-14% dotriacontanol (C-32), and 4-5% tetratriacontanol (C-34).

The monoester fraction in carnauba wax can is very similar to the monoester fraction of rice bran wax. The monoester fraction of rice bran wax comprises 87-98% of the total and carnauba wax monoesters accounts for 54-84% of the total. The predominant monoester carbon chain lengths have been reported to be C48-64 and C56 for both rice bran wax and carnauba wax, respectively. 


Table 3: Study comparison of Standard Valued for Rice Bran Wax and Carnauba Wax*

Observed values of
Rice Bran Wax
Standards value of
Carnauba Wax
Solubility
Insoluble in water, soluble in ether, ethanol
and isopropyl alcohol

Insoluble in water,
soluble in ether, ethanol

 Melting point
80.5 °C
78- 88°C
Specific Gravity
0.912
0.990-.0999 at 25°C
Moisture content 
0.074 %w/w
019% w/w**
Saponification value
80.88
78-95
Acid Value
2.848
NMT 12
Ester value
78.04
68-85
Hydroxyl value
19.62
---
Unsaponifiable matter
40% w/w
50-55 w/w%
Iodine value
10
5-14
*Taken directly from Maru et al. 2012
**This value is copied directly from the article.; however, it is likely a typo.  

3.0       HAZARD ASSESSMENT 

Studies conducted with rice bran wax oxidized have been summarized below and include an acute oral toxicity study, a dermal irritation study, an eye irritation study, and a dermal sensitization study. Rice bran wax oxidized was also tested in an Ames assay. No subchronic or chronic studies are available for RBWO. Although there are limited long term studies available for rice bran wax oxidized, various studies have been conducted on surrogate chemicals (e.g., rice bran wax, carnauba wax, D-002, D-003, docosanol, docosanoic acid, and policosanol) which were used to make a safety finding on RBWO. The data from these studies, conducted on mice, rats, rabbits, and dogs over various time periods, have also been summarized below. 
 3.1    Acute Toxicity 

Acute Oral Toxicity 
Wistar (Crl:WI Han) rats (3 female/dose) were administered 2000 mg/kg of rice bran wax oxidized (in 1% aqueous carboxymethyl cellulose) via gavage. No mortalities occurred at 2000 mg/kg; therefore, the LD50 > 2000 mg/kg. (Latour, 2016a)

Acute Dermal Toxicity 
No acute dermal toxicity studies are available for rice bran wax oxidized. 

Acute Inhalation Toxicity
No acute inhalation toxicity studies are available for rice bran wax oxidized. 

Dermal Irritation 
Rice bran wax oxidized was tested for potential skin irritation. EPISKIN Small Model(TM), a three-dimensional human epidermis model, was used. The mean absorption at 570 nm was 0.840 for the negative control, 0.070 for the positive control, and 0.608 for the test material. Viability was 8% for the positive control and 72% for the test material. Since the mean relative tissue viability was above 50% after the 15-minute exposure period, the test material is considered non-irritant. (Eurlings, 2015a)
Eye Irritation 
Rice bran wax oxidized was tested in an in vitro bovine corneal opacity and permeability assay. The mean in vitro irritation score was 0 for the negative control, 149.5 for the positive control and 2.8 for the test material after 4hrs of treatment. Based on the results of this study, rice bran wax oxidized did not cause eye irritation to the bovine cornea by either measurement of opacity or permeability. (Eurlings, 2015b)
Dermal Sensitization
Rice bran wax oxidized was tested in a local lymph node assay (LLNA) using female CBA/J mice (5/group) at doses of 0 (methyl ethyl ketone) 2, 5 and 10% (w/w). No irritation was observed and all lymph nodes appeared normal. Body weights and body weight gain of treated mice were similar to controls. The mean disintegration per minute (DPM)/animal values were 756, 1006, 1015 and 948 for the control, 2, 5 and 10% groups, respectively. The mean stimulation index (SI) was < 3 (i.e., 1.3 for all doses), therefore, rice bran wax oxidized was not considered to be a dermal sensitizer. (Latour, 2016b)
3.2    Subchronic Toxicity  

A 90-day gavage study with D-003 was conducted on male and female Sprague-Dawley rats (12/sex/group) at dose levels of 0, 50, 500, or 1250 mg/kg/day. No adverse effects of treatment were observed. One death occurred in a female in the 500 mg/kg/day group which was attributed to a dosing accident (i.e., test substance was found in the lungs). There were no signs of clinical toxicity observed and no significant differences were seen in body weights, hematology, clinical chemistry parameters, and organ weights. In addition, no treatment-related macroscopic or microscopic changes were observed. The No Observed Adverse Effect Level (NOAEL) was 1250 mg/kg/day (highest dose tested). (Gamez et al., 2000)

A 90-day gavage study was conducted with D-002 in male and female Sprague-Dawley rats (12/sex/group). The test material was administered as a suspension in acacia gum/water (10 mg/ml) at dose levels of 0, 5, 25, 125, or 625 mg/kg/day. There were six deaths in the study which upon necropsy were found to be related to animal manipulation errors during gavage and not treatment related. There were no treatment-related effects on body weight gain, food consumption, hematology, clinical chemistry parameters or organ weights. In addition, no microscopic effects were seen. The NOAEL was 625 mg/kg/day (highest dose tested). (Roderio et al., 1998)

3.3     Chronic

	Carnauba wax
Male and female Beagle dogs (6/sex/group) were fed diets containing 0, 0.1, 0.3 or 1% carnauba wax (i.e., 25, 75, or 250 mg/kg/day) for 28 weeks. Body weight, food consumption, hematology, clinical chemistry and urinalysis values and organ weights of animals fed carnauba wax were comparable to those of control dogs. No findings were observed upon ophthalmic examinations nor were there any treatment-related effects observed in gross or histopathological examinations. The NOAEL was 250 mg/kg/day (highest dose tested). (Parent et al., 1983a)

Docosanol
Male and female CD rats (20/sex/dose) were administered docosanol by gavage in 1% w/w aqueous Tween 80 at dose levels of 0, 10, 100, or 1000 mg/kg/day for 26 weeks. One male rats died in the 100 mg/kg/day group during study week 24.  The death was attributed to a dosing error.  No effects were observed on body weight, body weight gain, food conversion efficiency, organ weights, hematology, clinical chemistry or urinalysis parameters, ophthalmology or bone marrow at any dose tested. No treatment related findings were observed at necropsy or upon microscopic examination of tissues. The NOAEL was 1000 mg/kg/day (highest dose tested). (Inglesias et al., 2002)

Beagle dogs (4/sex/dose) were given docosanol by gavage in 1% w/w aqueous Tween 80 at dose levels of 0, 20, 200 or 2000 mg/kg/day for 27 weeks. There were no effects on general condition, body weight, food consumption, relative organ weights, ophthalmoscopy, macroppahology, or histopathology. Clinical signs were limited to pale feces in all dogs treated with 2000 mg/kg/day. The occurrence of pale feces is indicative of unabsorbed long-chain fatty alcohol. The NOEAL was 2000 mg/kg/day (highest dose tested). (Inglesias et al., 2002)
	D-003
Male and female Beagle dogs (3/sex/group) were administered D-003 by gavage at dose levels of 0 (aqueous gum acacia), 200, or 400 mg/kg/day for 9 months. There were no deaths or clinical signs of toxicity in the study. Body weights of the dogs were unaffected by treatment. In addition, no treatment-related effects were observed on ophthalmology, hematology parameters, organ weights, or on macroscopic or microscopic examination of tissues/organs. The only clinical chemistry parameters affected by treatment were triglyceride and total cholesterol levels which are known pharmacological effects of D-003. Effects on platelet aggregation and bleeding time were observed, but not on coagulation parameters. The effects on platelet aggregation and bleeding time are also expected pharmacological effects. Consequently, it was considered that D-003 was not toxic to dogs at the doses tested. The NOAEL was 400 mg/kg/day (highest dose tested). (Gamez et al., 2004)
Male and female OFI mice (50/sex/dose) were administered D-003 by gavage six days/week for 18 months at dose levels of 0, (10% aqueous gum acacia), 50, 500 or 1500 mg/kg/day. No difference in mortality was observed among the groups including the control group and there were no clinical signs of toxicity. Body weights and food consumption were similar among the control and treated groups throughout the study. No significant effects were seen on mortality, bodyweight gain, food consumption, hematology, clinical chemistry parameters, absolute and relative organ weights, and tumor incidences. There was no increase in the frequency of neoplastic and non-neoplastic tumors. Of the 41 deaths that occurred during the study, 9 were due to neoplastic lesions which were without difference in incidence between groups. D-003 was not carcinogenic to OFI mice administered over a period of 18 months under the conditions of this study. The NOAEL was 1,500 mg/kg/day (highest dose tested). (Noa et al., 2009)
 - 
Male and female Sprague-Dawley rats (60/sex/group) were administered D-003 at dose levels of 0 (10% aqueous gum acacia), 50, 500 and 1,500 mg/kg/day by gavage, five days/week for 24 months. There were no treatment-related effects on morality, clinical signs of toxicity, body weights, food consumption, hematology parameters, clinical chemistry parameters (except for a reduction in triglycerides), organ weights or macroscopic or microscopic findings. Glomerulonephrosis, the most frequent non -neoplastic lesion observed, occurred in 9 control males (15%) and 1 (1.7%), 5 (8.3%) and 7 (11.7%) males treated with 50, 500 and 1,500 mg/kg/day D-003, respectively and in 6 controls females (10%) and 1 treated (500 mg/kg/day) female (1.7%). No increase in any tumor type was observed. The NOAEL was 1,500 mg/kg/day. D-003 was not carcinogenic in rats under the conditions of this study. (Gamez et al., 2007)

	Policosanol
Policosanol was administered to male and female Sprague-Dawley rats (20/sex/dose) by gavage at dose levels of 0 (10% aqueous gum acacia), 0.5, 5, 50, or 500 mg/kg/day for 12 months. Treatment of rats for 12 months had no effect on mortality, body weights, food consumption, hematology or clinical chemistry parameters, organ weights or histopathology. The NOAEL was 500 mg/kg/day. (Aleman et al. 1994b)

Beagle dogs (4/sex/dose) were treated with policosanol by gavage for 52 weeks at dose levels of 0 (10% aqueous gum acacia), 30, or 180 mg/kg/day. There were no effects on body weight gain, histopathology, or blood chemistry. Total serum cholesterol, but not triglyceride or high-density lipoprotein-cholesterol, was statistically significantly decreased at both dose levels at each time point evaluated. This is a known pharmacological effect. The NOAEL for policosanol in dogs was 180 mg/kg/day (highest dose tested). (Mesa et al., 1994)

Male and female Sprague-Dawley rats (55/sex/dose) were administered policosanol by gavage daily at dose levels of 0 (acacia gum-water), 50, or 500 mg/kg/day for 24 months. Survival, body weight gain, organ weights and the incidence of non-neoplastic and neoplastic lesions were unaffected by treatment. The NOAEL was 500 mg/kg/day (highest dose tested). Policosanol was not carcinogenic when administered to Sprague-Dawley rats in this study. (Aleman et al., 1994a)

In an 18-month mouse carcinogenicity study, male and female Swiss mice were administered policosanol by gavage at dose levels of 0 (10% aqueous gum acacia), 50, or 500 mg/kg/day. Treatment of mice for 18 months had no effect on mortality, clinical observations, body weights, food consumption, organ weights or on the incidence of neoplastic or non-neoplastic lesions. The NOAEL was 500 mg/kg/day (highest dose tested). (Aleman et al., 1995)

	D-002
Groups of male and female Sprague-Dawley rats were administered D-002 for one year by gavage at dose levels of 0 (10% aqueous gum acacia), 250, 500, or 1000 mg/kg/day. Seven males and six females in various dose groups, including one female and one male in the control group, died during the study. The journal article outlining the study (Roderio et al., 1998) states that all deaths were due to gavage errors. "All of them related with the animal manipulation during the gastric entubation (sic)". There were no statistically significant effects on body weights or food consumption.  In addition, no effects were observed on hematology, clinical chemistry parameters, or organ weights. There was also no evidence of increases in the incidence of any microscopic lesions.  The NOAEL was determined to be 1000 mg/kg/day (the highest dose tested). (Roderio et al., 1998)

D-002 was administered to male and female beagle dogs (4/sex/group) at levels of 0 (gum acacia), 50 or 250 mg/kg/day by gavage for one year. No signs of toxicity were observed throughout the study. No mortality occurred and there was no effect on weight gain or food consumption. No hematological, blood biochemical or histopathological effect were seen. The NOAEL in this study was 250 mg/kg/day. (Aleman et al., 2001)
 3.4    Reproductive and Developmental Toxicity
	
      Carnauba wax 
Male and female Wistar (Harlan/Wistar HAN) rats (25/sex/group) were administered diets containing 0, 0.1, 0.3 or 1% carnauba wax four weeks prior to and throughout mating. Females were also fed throughout gestation and lactation. Twenty-five pups per sex for each dose where randomly selected to take part in a 13-week feeding study. According to the journal article reviewing the study, "Mean compound intake calculated on the basis of food consumption over the full 13-wk study was 0.08, 0.25, and 0.81 g/kg/day for males and 0.09, 0.27 and 0.67 g/kg/day for females treated at levels of 0.01, 0.3 and 1.1% carnauba wax, respectively". There were no effects of treatment on clinical signs, mortality, body weights or food consumption. In addition, no effects were observed on the reproductive parameters including fertility, gestation, viability, lactation indices, and pup weights. The maternal and reproductive toxicity NOAEL in this study is the highest dose tested (i.e., 1% dietary carnauba wax equivalent to 810 g/kg/day in males and 670 g/kg/day in females). 

In the subchronic portion of the study, body weights and food consumption were not affected by treatment. Statically significant increase in hematocrit in females (low and high dose) and a statically significant decreased of serum glutamate-pyruvate transaminase in males only (two higher doses) but without a dose response. Free fatty acid levels were statistically decreased at the two highest dietary levels as well. The decrease in males was dose related; however, it was not in females.  The significance of this finding is unclear considering one of the components of carnauba wax is long chain fatty acids. The NOAEL was considered by the review author to be the highest dose tested (i.e., 810 mg/kg/day in males and 670 mg/kg/day in females). (Parent et al., 1983b)

Docosanoic acid 
A summary of a study on docosanoic acid described in the European Chemicals Agency (ECHA) database as a result of the Registration, Evaluation, Authorization, and Restrictions of Chemical (REACH) dossier describes a study of male and female Crj:CD(SD) rats (13/sex/group) administered docosanoic acid by gavage at dose levels of 0 (corn oil), 100, 300, or 1000 mg/kg/day. Animals were dosed from 14 days prior to mating to day 3 of lactation for females and for males, from 14 days prior to mating, through mating and after for a total of 42 days. No treatment related clinical signs of toxicity or mortality were observed.  No effects were observed on body weights and body weight gain. No gross or microscopic treatment-related abnormalities were observed. Reproductive parameters and indices were unaffected by treatment. Additionally, there were no adverse effects on pups, including on sex ratio, body weights or viability. No external abnormalities were observed in the any of the treated groups nor were any abnormalities observed upon gross necropsy of the pups. Effects were observed on some hematology and clinical chemistry parameters and on organ weights. According to the summary, significant decreases in MCHC were observed in male rats at 300 and 1000 mg/kg/day but in the absence of any other effects on blood parameters or effects on females. Glucose levels were significantly decreased in male rats at 1000 mg/kg/day and alkaline phosphatase level test (ALP) was significantly decreased at all dose levels in males.  None of the conditions typically associated with decreased ALP were reported. Absolute and relative liver weight was significantly decreased in male rats and absolute kidney weight was significantly decreased in female rats at 1000 mg/kg/day. The observed decreases in absolute and relative organ weights were not considered of toxicological significance because no histopathology of the kidney or liver was observed. No raw data were presented in the ECHA summary, so the Agency is unable to determine if these effects are truly adverse. The effects on the liver and kidney may have been within historical controls or may not have shown a clear dose response. The study author states that "no treatment related adverse effects were found in either dose group 0, 100, 300, or 1000 mg/kg/day", and considered the NOAEL to be 1000 mg/kg/day for systemic, reproductive and developmental toxicity. The Agency agrees that a NOAEL of 1000 mg/kg/day (highest dose tested) is appropriate for reproductive and developmental toxicity based on the information in the dossier. Based on the lack of effects seen in other studies the Agency feels that the effects mentioned in this study may not be representative of effects of toxicity and may in fact be an adaptive response. 

D-003
In a developmental toxicity study, pregnant female Sprague-Dawley rats (25/dose) were administered D-003 by gavage at dose levels of 0 (1% gum acacia), 5, 100, or 1000 mg/kg/day from gestation day 6 through 15. No maternal clinical signs of toxicity were observed or were there any effects on maternal body weight or body weight gain. No effects were observed on reproductive parameters or on the incidence of external, visceral, or skeletal findings in fetuses. The maternal and developmental toxicity NOAEL was 1000 mg/kg/day (highest dose tested). (Rodrequez et al., 2003)

In a developmental toxicity study, 27 mated female New Zealand white rabbits were administered D-003 by gavage at dose levels of 0 (1% gum acacia), 500, or 1000 mg/kg/day from gestation day 6 through 18. Body weights and body weight gain of the dams were unaffected by treatment. No effects on reproductive parameters, including fetal body weights, were observed. The incidence of external, visceral and skeletal abnormalities was similar in the treated and control groups. The maternal and developmental NOAEL in the study was 1000 mg/kg/day (highest dose tested). (Rodriguez et al., 2004)
	D-002
Groups of pregnant Sprague-Dawley rats were administered D-002 by gavage at dose levels of 0 (10% aqueous gum acacia),100, 320, or 1000 mg/kg/day from gestation days 6 through 15. No clinical signs of toxicity were observed. Reproductive parameters were unaffected by treatment. The numbers of implantations, corpora lutea, sex ratio, resorptions, live fetuses and fetal weight were comparable to controls. No treatment-related fetal abnormalities were observed. The NOAEL for maternal and developmental toxicity was 1000 mg/kg/day (highest dose tested). (Rodriguez, Gamez, Sanchez, & Garcia, 1998)

Groups of 16 to 20 pregnant New Zealand white rabbits were administered D-002 by gavage at dose levels of 0 (10% gum acacia), 100, 320, or 1000 mg/kg/day from gestation days 6 to 18. No clinical signs of toxicity were observed in the dams and there were no effects on body weight or body weight gain. In addition, reproductive parameters were unaffected by treatment and no treatment-related fetal abnormalities were observed. There was no effect on implantations, resorptions, litter size, sex ratio or fetal body weight. The NOAEL for maternal and developmental toxicity was 1000 mg/kg/day (highest dose tested). (Rodriguez, Gamez, Sanchez, & Garcia, 1998)

      Policosanol
In a three part teratogenic and reproductive study (Rodriguez & Garcia, 1994), rats and rabbits were exposed to policosanol via gavage. Policosanol was administered to pregnant Sprague-Dawley female rats (25/dose) at dose levels of 0 (10% aqueous gum acacia), 5, 50, or 500 mg/kg/day from day 6 to day 15 of gestation to determine the potential for developmental toxicity. No effects were observed on maternal or fetal body weights or on the mean number of corpora lutea, implantations, resorptions, or dead and live fetuses. In addition, no statically significant treatment-related visceral or skeletal findings were observed. The NOAEL for maternal and developmental toxicity in the rat was 500 mg/kg/day (highest dose tested). 

Policosanol was also administered to pregnant New Zealand white rabbits (15/dose) at dose levels of 0 (10% aqueous gum acacia), 500, or 1000 mg/kg/day from gestation day 6 through 18. No effects were observed on maternal or fetal body weights or on the mean number of corpora lutea, implantation, resorptions or dead and live fetuses. No statistically significant treatment-related external, visceral or skeletal findings were observed. The NOAEL for maternal and developmental toxicity in the rabbit was 1000 mg/kg/day (highest dose tested). (Rodriguez & Garcia, 1994)

In order to further evaluate fertility and reproductive toxicity, policosanol was given to female Sprague Dawley rats for two weeks prior to mating and throughout mating, pregnancy and lactation to postnatal day 21. It was also give to male rats for 60 days before mating and during mating at levels of 0 (10% gum acacia), 5, 50, or 500 mg/kg/day policosanol. Fertility was unaffected by treatment. In addition, no effects were observed on reproductive parameters in the females. There was no difference in the number of corpora lutea, implantations, resorptions, dead or live fetuses, preimplantation loss, postimplantation death and fetal weights. The reproductive toxicity NOAEL was 500 mg/kg/day (highest dose tested. (Rodriguez & Garcia, 1994)

A two-generation peri- and post-natal gavage toxicity study was conducted with policosanol in female Sprague-Dawley rats (16/dose) at dose levels of 0, 5, 50, or 500 mg/kg/day from day 15 of pregnancy to day 21 after parturition. No significant differences were observed on litter size, survival through weaning period, sex ratio, and body weight. No adverse effects were seen in the postnatal growth, behaviors, or reproductive ability of pups. One male and one female from each litter were selected at random to produce the second generation (F2) of pups. There were no significant differences in the F2 generation in regard to number of pups born, postnatal pup survival, sex ratio, and mean pup weights. In addition, there were no effects on physical or behavioral development of the F2 generation. The NOAEL for maternal and offspring toxicity was 500 mg/kg/day (highest dose tested). (Rodriguez & Garcia, 1998)

Policosanol was tested in a 3-generation gavage reproductive toxicity study in male (15/dose) and female (30/dose) Sprague-Dawley rats at dose levels of 0, 5, 50 or 500 mg/kg/day. For subsequent mattings 20 females and 10 males were used. For each generation, two litters were reared until they were 3 weeks old. No treatment related clinical signs were recorded in pregnant or lactating mothers. In subsequent generations there were no differences among groups in the number of animals conceived, number of pups born, sex ratio, number of pups that survived until weaning, and pup body weight. There was also no effect on behavior or physical appearance. The NOAEL for maternal, reproductive, and developmental toxicity was 500 mg/kg/day (highest dose tested). (Rodriguez, Sanchez, & Garcia, 1997)
 3.5    Neurotoxicity

No neurotoxicity studies were conducted on rice bran wax oxidized or any of the structurally similar chemicals. Although no specific neurotoxicity studies were conducted there was no evidence of neurotoxicity in any of the acute, subchronic, chronic, or developmental studies; therefore, the agency does not expect rice bran wax oxidized to be neurotoxic. 
 3.6    Mutagenicity/Cytotoxicity 

Reverse Gene Mutation 
Rice bran wax oxidized was tested in the Ames Salmonella assay (plate incorporation and pre-incubation assays) in Salmonella strains TA1535, TA1537, TA100 and TA98 and E. coli strain WP2uvrA at concentrations of 0, 17, 52, 164, 512 and 1600 ug/plate in the presence and absence of metabolic activation. When tested up to its limit of solubility, rice bran wax oxidized did not induce reverse mutations in any strain tested with or without metabolic activation and was considered negative in the Ames Salmonella and E. Coli reverse mutation assays. (Verspeek-Rip, 2015)    

D-003 was tested in the Ames Salmonella assay (plate incorporation method and the preincubation method) using tester strains TA98, TA100, TA1535, TA1537 and TA1538.  In both experiments concentrations of 0 (vehicle not indicated), 5, 50, 500, 2000 and 5000 ug/plate were tested both with and without metabolic activation. No increase in reverse mutations was observed in any test with or without metabolic activation using either the plate incorporation or preincubation methods. D-003 was negative in the Ames Salmonella assay. (Gamez, Roderio, Fernandez, & Acosta, 2002)

An Ames Salmonella assay (plate incorporation test) was conducted on docosanol (behenyl alcohol) using tester strains TA1535, TA1537, TA1538, TA98 and TA100 with and without metabolic activation.  Docosanol was negative both with and without metabolic activation. The positive controls produced the expected response. (Inglesias et al., 2002)

An Ames Salmonella assay (plate incorporation test) was conducted on policosanol using tester strains TA1535, TA1537, TA1538, TA98 and TA100 with and without metabolic activation. Policosanol was negative both with and without metabolic activation. (Rodriguez, Alfonso, & Acosta, 2006)

Micronucleus Assay
Male and female NMRI mice (6/sex/dose) were administered D-003 by gavage for 6 consecutive days at 0 or 2000 mg/kg and their bone marrow was evaluated for the presence of micronuclei. Treatment of mice with D-003 did not result in an increase in either micronuclei/PCE or PCE/NCE. D-003 was considered negative in the mouse micronucleus assay. (Gamez et al., 2001)

Docosanol was evaluated for its ability to induce micronuclei in the bone marrow of NMRI mice (5/sex/group) at a single gavage dose of 0 (polyethylene glycol), 50, 150 or 500 mg/kg. Docosanol did not increase the frequency of bone marrow polychromatic erythrocyte (PCE) micronuclei in mice in vivo. (Inglesias et al., 2002)

Alkaline Comet Assay
Five Male Sprague-Dawley rats were administered D-003 by gavage at doses of 0 (acacia gum/water) or 1250 mg/kg for 90 days. A cell suspension was made from a small piece of liver from each animal and used for the Comet assay. D-003 did not result in DNA damage. No single strand breaks or alkali-labile site were introduced in isolated liver cells. D-003 was negative in the alkaline Comet assay. (Gamez et al., 2001)

Chinese Hamster Cell in vitro 
Docosanol was tested for its ability to produce gene mutations in Chinese hamster V79 cells at concentrations of 0 (ethanol), 2.0, 7.5, 15.0 and 20.0 ug/mL with and with metabolic activation.  Docosanol was negative with and without metabolic activation in this test system. (Inglesias et al. 2002)

Docosanol was tested for its ability to induce chromosomal aberrations in Chinese hamster V79 cells at concentrations of 0, 0.6, 10 and 20 ug/mL with and without metabolic activation.  Docosanol did not induce the incidence of chromosomal aberrations at the concentrations tested, at any of the three treatment times or with or without metabolic activation. (Inglesias et al. 2002)
 3.7    Carcinogenic Potential 

Various long-term studies have been conductive on surrogates of rice bran wax oxidized. There was no evidence of increased neoplastic or non-neoplastic lesions. Similarly, all mutagenicity studies on rice bran wax oxidized or acceptable surrogates were negative which makes it unlikely that rice bran wax oxidized would be carcinogenic.

3.8    Endpoint Selection and Levels of Concern

Available toxicity studies on rice bran wax oxidized and various surrogate chemicals indicate that rice bran wax oxidized has a very low acute and chronic toxicity. With the exception of one reproductive/developmental study on docosanoic acid, no adverse effects of treatment were seen in any of the studies in mice, rats, rabbits, and dogs at any dose tested up to the limit dose of 1000 mg/kg/day. In the study on docosanoic acid some statically significant effects were seen in hematology and clinical chemistry parameters and organ weights. However, only the REACH summary was available, the full data set was not provided for this study so the Agency is unable to determine if the effects seen were truly adverse with a clear dose-response or if they were within historical controls. The REACH document gave the NOAEL for systemic, reproductive, and developmental toxicity as 1000 mg/kg/day (highest dose tested). 

Considering the weight of evidence and the lack of an endpoint in all other studies, this is study is not considered as reliable for use in evaluating toxicity although the reproductive and developmental toxicity portion of the study showed no effect of treatment on reproductive and developmental parameters. No adverse effects were seen in any of the other studies, therefore, no endpoint of concern has been identified for any of the acute, subchronic, or chronic studies up to and beyond the limit dose of 1000 mg/kg/day. Since no endpoint of concern was identified a quantitative risk assessment for rice bran wax oxidized is not necessary.
   

3.9    FQPA Safety Factor Considerations

FFDCA Section 408(b)(2)(c) provides that EPA shall 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 completeness of the database on toxicity and exposure unless EPA determines based on reliable data that a different margin of safety will be safe for infants and children. This additional margin of safety is commonly referred to as the FQPA safety factor (SF). In applying this provision, EPA either retains the default value of 10X, or uses a different additional safety factor when reliable data available to EPA support the choice of a different factor. 

The available acute and repeat dose toxicity studies on rice bran wax oxidized and surrogate chemical data indicate that rice bran wax oxidized has low toxicity with no adverse effects of treatment to seen at the limit dose of 1000 mg/kg/day. There is no indication, based upon the available data, that rice bran wax oxidized is neurotoxic or immunotoxic and therefore, would not result in increased susceptibility in infants or children. 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. Taking into consideration all available information, there is no concern, at this time, for increased sensitivity to infants and children to rice bran wax oxidized when used as inert ingredient in pesticides formulations.

4.0       EXPOSURE ASSESSMENT

4.1    Dietary Exposure

Although dietary exposure to rice bran wax oxidized may occur from eating foods treated with pesticide formulations containing this inert ingredient and drinking water containing runoff from soils containing the treated crops, no endpoint of concern was identified and therefore, a quantitative dietary exposure assessment for rice bran wax oxidized was not conducted. 

4.2    Cancer Exposure 

Surrogate chemical data submitted to support rice bran wax oxidized did not show any evidence of carcinogenicity in various chronic studies. In addition, rice bran wax oxidized and surrogate chemicals tested negative in mutagenicity assays and no evidence of specific target organ toxicity was observed in the repeat dose studies. Based on the available toxicity data, rice bran wax oxidized is not expected to be carcinogenic. Therefore, a cancer dietary exposure assessment is not necessary to assess cancer risk. 

4.3    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, indoor pest control, termiticides, and flea and tick control on pets). The proposed use of rice bran wax oxidized under 40 CFR 180.910, 180.930, and 180.940(a) could result in residential exposures to rice bran wax oxidized; however, there are no toxicological effects of concern in available studies and therefore, it is not necessary to conduct a quantitative assessment of residential (non-occupational) exposures and risks. 

4.4    Occupational Exposure 

The Agency has reviewed the available toxicological information for rice bran wax oxidized and there are no substantiated adverse toxicological effects observed in the studies submitted up to the limit dose of 1000 mg/kg/day, therefore a quantitative occupational risk assessment is not necessary. 

5.0       AGGREGATE RISK 

Because no substantiated adverse effects, attributable to a single or repeat exposure, is seen in the toxicity database for the rice bran wax oxidized or its surrogates a qualitative risk assessment was conducted and subsequently, it is not necessary to aggregate dermal and inhalation residential exposures with estimated dietary exposures. 

6.0       CUMMULATIVE EXPOSURE 

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 rice bran wax oxidized to share a common mechanism of toxicity with any other substances, and rice bran wax oxidized does do not appear to produce a toxic metabolite produced by other substances. For the purposes of this tolerance action, therefore, EPA has assumed that the rice bran wax oxidized 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. 
 7.0       ENVIRONMENTAL FATE 

In determining the environmental fate of rice bran wax oxidized the Agency considered physical/chemical properties (Table 1) and studies submitted. 

The substance is soluble in water (< 1g/L) and could exist in both the vapor or particulate phase (VP 1 x 10-5 p/hPa). A Ready Biodegradability Modified Sturm Test was conducted on rice bran wax oxidized. A carbon content (TOC) of 10.8 mg C/L was used in the two test vessels. Colony forming units (CFU) of the inoculum corresponding to approximately 1.04 x 107 CFU/L were determined by standard dilution plate count. The mean degradation after 28-days was 25%, indicating that the substance is not readily biodegradable. (47) (Maischak, 2016)

Rice bran wax oxidized is derived from a naturally occurring substance. The Agency does not expect that use as an inert ingredient will significantly add to the overall natural presence of RBW.



 8.0      ECOLOGICAL EFFECTS
 8.1   Fish Toxicity 

Freshwater and Saltwater Fish Acute Toxicity Test
Zebra fish (Danio rerio) were exposed to 0, 5, 10, or 15 mg/L of docosanoic acid of for 96 hours under static conditions. No mortality or clinical signs of toxicity occurred. The LC50 was >15 mg/L. (ECHA Dossier 15830/6/2/2)
 8.2   Invertebrate Toxicity 
Daphnia magna 
Four replicates of Daphnids (5/dose/replicate) were exposed to 0 (dilution water), 6.25, 12.5, 25, 50 or 100 mg/L saturated solution of rice bran wax oxidized for 48 hours under static conditions. Total organic content (TOC) did not vary significantly from start to end of exposure. The EC50 at both 24 and 48 hours was >100 mg/L based on nominal (percent saturated solution).  (Scheerbaum, 2016a)

8.3    Algal Toxicity

Alga Growth Inhibition Test
A saturated solution of rice bran wax oxidized was prepared and tested at 0, 10.0, 17.8, 31.6, 56.2 and 100% for growth rate and yield effects on fresh water green alga, Pseudokirchneriella subcapitata. The saturated solution was prepared with a nominal concentration of 100 mg test material/L. The total organic carbon did not change over the course of the 72-hour study. No effects on growth rate were observed at 10.0, 17.8 and 31.6% of the saturated solution. At 58.2 and 100% saturation, growth rate was inhibited 10 and 26%, respectively. Yield was unaffected at 10.0 and 17.8% saturated solution. At 31.6, 56.2 and 100% saturated solution, yield was inhibited 9, 42 and 77%, respectively. The No Observed Effect Concentration (NOEC) and Lowest Observed Effect Concentration (LOEC) for growth rate and yield were reported as 31.6 and 56.2% saturated solution, respectively. The 72-hour ErC50 was >100% saturated solution and the EyC50 was 61.9% saturated solution. (Scheerbaum, 2016b)

8.4    Terrestrial Toxicity
 
A mixture of long chain fatty alcohols was evaluated in a 1984 study by Menon and Srivastava. Mixtalol, contains long chain fatty alcohols (C24 to C34). In the first experiment, seeds were soaked with Mixtalol which resulted in a statistically significant increase in root length and number of laterals for paddy seedlings, a statistically significant increase in root and shoot length for wheat, and a statistically significant increase in root length and number of laterals for maize. Mixtalol (1 ppm) sprayed on paddy seedlings 4 days after germination resulted in a statistically significant increase in root length. Shoot fresh weight of wheat seedlings sprayed with 1.5 ppm Mixtalol increased 8.2 to 10.3% compared to controls, dry shoot weight increased 10.6 to 12.5 % and root dry weight 23.0 to 25.1%.  There were no adverse effects of Mixtalol noted in any of these experiments. (Menon and Srivastava, 1984)

In another study, seedlings of green gram (Vigna radiate L. Wileczek; mung beans) were sprayed with triacontanol at concentrations of 0, 0.5, 1.0 and 2.0 mg dm-3 15 and 25 days after planting. Root length was statistically increased at concentrations of triacontanol up to 1.0 mg dm[-3] when treated 15 or 25 days after planting. Shoot length was statistically increased 25 days after treatment at 0.5 mg dm[-3] triacontanol compared to controls. Plant height was also statistically increased at 0.5 and 1.0 mg dm[-3] triacontanol in plants sprayed 15 days after planting and at 0.5 mg dm[-3] when sprayed 25 days after planting.  Root fresh mass was statistically increased in plants sprayed 15 and 25 days after planting with 0.5 mg dm[-3] triacontanol.  Shoot fresh mass was statistically decreased after spraying with 1.0 or 2.0 mg dm[-3] triacontanol. Similar effects were observed on plant fresh mass. (Jones, Wert, and Ries, 1979)

9.0       RISK CHARACTERIZATION   

To make rice bran wax oxidized, RBW is chemically alter by partial cleavage of the esters and oxidation of the resulting alcohols leading to the formulation of carboxylic acid. RBW is a hard, crystalline vegetable wax obtained from the oil obtained from rice husks. Rice bran wax oxidized is currently used in agricultural applications (e.g. seed coatings, fertilizer coatings, adjuvants), polish and care applications (e.g. shoe, floor, and car care), wax emulsions and dispersions, coating and inks, textiles, leather, and paper. Under the proposed use pattern, rice bran wax oxidized will be used as an inert ingredient in pesticide formulations applied pre-and post-harvest, to animals, and in antimicrobial formulations (food-contact surface sanitizing solutions). According to the submitter, rice bran wax oxidized will be used as an inert ingredient in agricultural formulations as a flow aid, surface protection, film-forming, carrier, coating agent, and adjuvant. 

Rice bran wax oxidized consists mainly of long chain fatty acids, alcohols, and esters. According to the GRAS notification for rice bran wax, the potential for absorption of rice bran wax via the gastrointestinal (GI) tract is limited. The long-chain fatty acid esters present in plant-based waxes are generally thought to be poorly absorbed in the GI tract as uptake is thought to decrease as chain length and hydrophobicity increase. Similarly, it is unlikely that RBWO would be systemically available. 

An acute study revealed low oral toxicity. When tested, the chemical was shown to be non-irritating to the skin and eyes and was not a skin sensitizer. Acute dermal and inhalations studies for rice bran wax oxidized were not available. 

With the exception of one reproductive/developmental study on docosanoic acid, no adverse effects of treatment were seen in any of the studies in mice, rats, rabbits, and dogs up to the limited dose of 1000 mg/kg/day. In the study on docosanoic acid some effects were seen in hematology and clinical chemistry parameters and organ weights. However, the full data set was not provided for this study so the Agency is unable to determine if the effects seen were truly adverse with a clear dose response or if they were within historical controls. Without the actual data to review the Agency feels that this study cannot be properly evaluated and it is therefore, not considered as reliable as the other studies for which data was presented. The REACH document gave the NOAEL for systemic, reproductive, and developmental toxicity as 1000 mg/kg/day which was the highest dose tested indicating that they did not see these effects as adverse. 

There was no evidence of carcinogenicity in any of the studies presented including chronic studies and studies on mutagenicity and cytotoxicity. In addition, no neuropathological changes or effects were reported in any of the studies. The agency does not believe RBWO will be carcinogenic or neurotoxic. 

No endpoint of concern was identified for any of the acute, subchronic, or chronic studies submitted. In addition, no endpoint of concern was determined in any of the studies up to the limit dose of 1000 mg/kg/day. Since no endpoint of concern was identified in acute, subchronic, or chronic studies and because rice bran wax oxidized is not expected to be absorbed by the body, a quantitative risk assessment for RBWO was not performed. 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. 

While residential and occupational exposure are also possible, there are no toxicological effects of concern in available studies and therefore, it is not necessary to conduct a quantitative assessment of residential (non-occupational) exposures and risks. Similarly, a quantitative occupational risk assessment is not necessary. 

Based on ecological and fate data the Agency concluded that RBWO does not pose an ecological risk to terrestrial or aquatic species. Rice bran wax oxidized is made from a naturally occurring substance and its use as an inert ingredient in pesticide formulations is unlikely to add to the overall load of RBW in a way that would be toxic to humans or to the natural environment. 

Based on the evaluated toxicity studies coupled with the expected exposure from the use of this chemical as inert ingredient in pesticide products, EPA concludes that there is a reasonable certainty that no harm will result to the general population, including infants and children, from aggregate exposure to residues of waxes and waxy substances, rice bran, oxidized when used as an inert ingredient in pesticide formulations under 40 CFR 180.910, 180.930, and 180.940(a).







                                 Bibliography
Aleman, C., Ferreiro, R., Puig, M., Guerra, I., Ortega C., and Capote, A. (1994a) Carcinogenicity of policosanol in Sprague-Dawley rats: A 24-month study. Teratogenesis, Carcinogenesis and Mutagenesis 14:239-249.

Aleman, C., Mas, R., Hemandez, C., Rodeiro, I., Cerejido, E., Noa, M., Capote, A., Menendez, R., Amor, A., Fraga, V., Sotolongo, V., and Jimenez, S. (1994b) A 12-month study of policosanol oral toxicity in Sprague-Dawley rats. Toxicology Letters 70:77-87.

Aleman, C., Puig, M., Elias, E., Ortega, C. Guerra, R., Ferreiro, R., and Brinis, F. (1995) Carcinogenicity of policosanol in mice: An 18-month study. Food and Chemical Toxicology. 33:573-578.

Aleman, C., Rodeiro, I., Noa, M., Menendez, R., Gamez, R., Hernandez, C., and Mas, R. (2001) One-year dog toxicity study of D-002, a mixture of aliphatic alcohols. Journal of Applied Toxicology. 21:179-184.

Eurlings, I.M.J. (2015a) In vitro skin irritation test with Licocare RBW 106 using a human skin model. WIL Research Europe B.V. The Netherlands. Project. ID No. 509742. Report dated: November 23, 2015. Unpublished.

Eurlings, I.M.J. (2015b) Evaluation of the eye hazard potential of Licocare RBW 106 using the bovine corneal opacity and permeability test (BCOP Test). WIL Research Europe B.V. The Netherlands. Project ID: 509741. Report dated: November 23,2015. Unpublished.

European Chemicals Agency (ECHA) REACH Dossiers:
      https://echa.europa.eu/es/registration-dossier/-/registered-dossier/15830/7/6/2
      https://echa.europa.eu/es/registration-dossier/-/registered-dossier/15830/6/2/2


Gamez, R., Mas, R., Noa, M., Menendez, R., Aleman, C., Acosta, P., Garcia, H., Hernandez, C., Amor, A., Perez, J., and Coicochea, E. (2000) Acute oral and subchronic toxicity of D-003 in rats. Toxicology Letters 118:31-41.

Gamez, R., Mas, R., Noa, M., Menendez, R., Garcia, H., Gonzalez, J., Perez, Y., and Coicochea, E. (2004) Effects of chronic administration of D-003, a mixture of sugar cane wax high molecular acids, in beagle dogs. Drugs Exptl. Clin. Res. 30:75-88.

Gamez, R., Noa, M., Mas, R., Mendoza, N., Pardo, B., Menendez, R., Perez, Y., Gonzalez, R.M., Gutierrez, A., Marrero, G., Coicochea, E., Garcia, H., and Curveco, D. (2007) Long-term carcinogenicity of D-003, a mixture of high molecular weight acids from sugarcane wax, in Sprague Dawley rats: A 24 months study. Food and Chemical Toxicology 45:2352-2358

Gamez, R., Rodeiro, I., Fernandez, I., and Acosta, P. (2002) Preliminary evaluation of cytotoxic and genotoxic potential of D-003: Mixture of very long chain fatty acids. Teratogenesis, Carcinogenesis, and Mutagenesis. 22:175-181.

Gamez, R., Gonzalez, J., Rodeiro, I., Fernandez, I., Aleman, C., Rodriguez, M., Acosta, P., and Carcia, B. (2001) In vivo genotoxic evaluation of D-003, a mixture of very long chain aliphatic acids. Journal of Medicinal Food 4:85-91.

Inglesias, G., Hlywka, J., Berg, J., Khalil, M., Pope, L., and Tamarkin, D. (2002) The toxicity of behenyl alcohol: I. Genotoxicity and subchronic toxicity in rats and dogs. Regulatory Toxicology and Pharmacology 36:69-79.

Jones, J., Wert, V. and Ries, S. (1979) Specificity of 1-triacontanol as a plant growth stimulator and inhibition of its effect of other long-chain compounds. Planta 144:277-28

Latour, J.E.H.M. (2016a) Assessment of acute oral toxicity with Licocare RBW 106 in the rat (acute toxic class method). WIL Research Europe B.V., Germany, Project ID 511335. Report dated: February 15, 2016. Unpublished.

Latour, J. (2016b) Assessment of skin sensitization to Licocare RBW 106 (wax, rice bran, oxidized) in the mouse (local lymph node assay). WIL Research Europe B.V. Project ID 511336. Report dated March 21,2016. Unpublished.

Maischak, H. (2016). Licocare RBW 106 FL TP Ready Biodegradability Modified Sturm Test. Noack Laboratorien GmbH, Germany. Study ID 150622CG/AST16668. Study dated February 10, 2016. Unpublished.

Maru, A., Surawase, R. and Bodhe, P. (2012) Studies on physico-chemical properties of rice bran wax and its comparison to carnauba wax. Int. J. Pharm. Phytopharmacol. Res. 1:203-207.

Menon, K. and Srivastava, H. (1984) Increasing plant productivity through improved photosynthesis. Proc. Indian. Acad. Sci. (Plant Sci.) 93:359-378.

Mesa, A., Mas, R., Noa, M., Hernandez, C., Rodeiro, I., Gamez, R., Garcia, M., Capote, A., and Aleman, C. (1994) Toxicity of policosanol in beagle dogs: one-year study. Toxicology Letters 73:81-90.

Noa, M., Gamez, R., Mas, A., Guitierrez, A., Mendoza, N., Pardo, B., Merrero, G., Curveco, D., Coicochea, E., Garcia, H., Perez, Y., and Gonzalez, R.M. (2009) Study of the long-term carcinogenicity potential of D- 003, a mixture of high molecular weight sugarcane wax acids, in mice. Food and Chemical Toxicology. 47:687- 692.

Parent, R., Cox, G., Babish, J.G., Gallo, M.A., Hess, F.G., and Becci, P.J. (1983a) Subchronic feeding study of carnauba wax in beagle dogs. Food and Chemical Toxicology. 21:85-87.

Parent, R.A., Re, T.A., Babish, J.G., Cox, G.E., Voss, K.A., and Becci, P.J. (1983b) Reproduction and subchronic feeding study of carnauba wax in rats. Food and Chemical Toxicology. 21:89-93.

Rodeiro, I, Aleman, C., Noa, M., Menendez, R., Mas, C., Hernandez, C., and Garcia, M. (1998) Preclinical oral toxicology in rats of D-002, a natural drug with antiulcer effects. Drug and Chemical Toxicology. 21:151-162.

Rodriguez, S., Alfonso, J. and Acosta, P. (2006) In vitro mutagenic evaluation of policosanol. Revista CENIC Ciencias Biologicas 37:9-12.

Rodriguez, M., Gamez, R., Gonzalez, J., Garcia, H., Acosta, C., and Coicochea, E. (2003) Lack of developmental toxicity of D-003: a mixture of long-chain fatty acids in rats. Food and Chemical Toxicology. 41:89-93.

Rodriguez, M., Gonzalez, J., Aleman, C., Rodeiro, I., Arango, E., Gamez, R., Valdes, H., Garcia, H., Coicochea, E., and Acosta, C. (2004) Evaluation of the reproductive and developmental toxicity of the D-003, a mixture of long-chain fatty acids, in rats and rabbits. Food and Chemical Toxicology 42:1977-1985.

Rodriguez, M., Gamez, R., Sanchez, M. and Garcia, H. (1998) Developmental toxicity of D-002 (a mixture of aliphatic primary alcohols) in rats and rabbits. Journal of Applied Toxicology. 18:313-316.

Rodriguez, M. and Garcia, H. (1994) Teratogenic and reproductive studies of policosanol in the rat and rabbit. Teratogenesis, Carcinogenesis, and Mutagenesis 14:107-113.

Rodriguez, M. and Garcia, H. (1998) Evaluation of peri- and post-natal toxicity of policosanol in rats. Teratogenesis, Carcinogenesis, and Mutagenesis 18:1-7.

Rodriguez, M., Sanchez, M. and Garcia, H. (1997) Multigeneration reproduction study of policonsanol in rats. Toxicology Letters 90:97-106.

Scheerbaum, D. (2016a) Licocare RBW 106. Acute immobilisation test to Daphnia magna, static, 48 hours. Noack Laboratorien GmbH. Study ID 150622CG/DA116668. Report dated June 8, 2016. Unpublished.

Scheerbaum, D. (2016b) Alga, growth inhibition test with Pseudokirchneriella subcapitata, 72 hours. Noack Laboratorien GmbH, Germany. Study ID 150622CG/SPO16668. Study dated March 4, 2016. Unpublished.

Schmidt, D. (2006) GRAS determination of rice bran wax for use in food. June 3, 2016. Retrieved on 6/21/18 from https://www.fda.gov/downloads/Food/IngredientsPackagingLabeling/GRAS/NoticeInventory/ucm516030.pdf 
       GRAS internal citations:

      European Food Safety Authority (EFSA). 2012a. Scientific Opinion on the evaluation of the substances currently on the list in the Annex to Commission Directive 96/3/EC as acceptable previous cargoes for edible fats and oils -Part III of III. EFSA Journal 20 12; 1 0(12):2984. 
      European Food Safety Authority (EFSA). 2012b. Scientific Opinion on the re-evaluation of carnauba wax (E 903) as a food additive. EFSA Joumal2012;10(10):2880. 

      Food and Drug Administration (US FDA). 2004. GRN No. 151. GRAS Notification for Ethanol (Ethyl Alcohol). Prepared by Frito-Lay, Inc.

      Hargrove, JL, Greenspan, P, Hartle, DK. 2004. Nutritional significance and metabolism of very long chain fatty alcohols and acids from dietary waxes. Experimental Biology and Medicine, 229(3), 215-226.

      Logan BK, Distefano S. 1998. Ethanol content of various foods and soft drinks and their potential interference with a breath-alcohol test. Journal of Analytical Toxicology 32:181-183.
      
Vali, S., Ju, Y-H., Kaimal, T., and Chern, Y-T. (2005) A Process for the Preparation of Food-Grade Rice Bran Wax and the Determination of Its Composition. JAOCS (Journal of the American Oil Chemists' Society. 82:57-64
	Vali internal citations:

      Iwama, F., and S. Maruta, Composition of the Hard Portion of Crude Rice Bran Wax, Kogyo Kagaku Zasshi 72:2605 - 2608 (1969).

      Yoon, S.H., and J.S. Rhee, Composition of Waxes from Crude Rice Bran Oil, J. Am. Oil Chem. Soc. 59:561 - 563 (1982).

      Belavadi, V.K., and D.N. Bhowmick, An Investigation of Rice Bran Oil Tank Settling, Ibid. 65:241 - 245 (1988).

      Ito, S., T. Shzuki, and Y. Fujino, Wax Lipid in Rice Bran, Cereal Chem. 60:252 - 253 (1983).

      Wang, Y., H. Diao, and P. Ni, Composition of Rice Bran Wax, Zhongguo Youzhi 23:39 - 41 (1998).

Verspeek-Rip, C. (2015) Evaluation of the mutagenic activity of Licocare RBW 106 in the Salmonella typhimurium reverse mutation assay and the Escherichia coli reverse mutation assay (plate incorporation and pre-incubation methods). WIL Research Europe B.V. The Netherlands. Project ID; 509740. Report dated: September 16, 2015. Unpublished.


