Hop Beta Acids PC Codes: 005115 DP Numbers: 417758 & 417759 EPA Fi le No.: 83623-G , -R, & -E Rev iew Ty pe: Residue Studies , Tier I Tox , Tier I Non-targets

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

OFFICE OF CHEMICAl SAFETY & POUUTIOI PIEVEITIOI 

### MEMORANDUM DATE: 12/04/2014 SUBJECT: Science Review in Support of the Registration of HopGuard TGAI, HopGuard Liquid , and HopGuard , Respectively Contain in g 48.02% , 16% , and 16% Hops Beta Acids Resin, a New Active Ingred ie nt. Review of Residue Studies, Tier I Toxicity and Tier I Non

-Target Organism Studies and Rational es.



### FROM:

TO: EPA Reg. No.: Pet ition No.: Decision Nos.: DP Nos.: Chemical Class: PC Codes: CAS No. MRID s: 83623-G, -R & -E 3E8217 484728 , 484725&484727 417759,417758,417760 Biochemical 005115 None Ass ig n ed 492519-0 I to -05 Russell S. Jones , Ph.D. , Senior Biologist 

### I s / 12 / 04/ 2014 Biochemical Pesticides Branch Biopesticides & Pollution Prevention Division (7511 P) Menyon Adams, Regulatory Action Leader 

### I s / 12/ 04/2014 Biochemical Pesticides Branch Biopesticides & Pollution Prevention Division (7511 P)



### ACTION REQUESTED The BetaTec Division of John I. Haas , Inc. requests registration ofthree products containing the new active ingredient , Hops Beta Ac id s Resin : A technical grade of the active in gredient (TGAI)Hop Beta Acids PC Codes: 0051 15 Review Type: Residue Studie s, Tier I T ox , Tier I Non-targets DP Numbers : 417758 & 4 1 7759 E PA File No.: 83623-G, -R, & -E

Hopguard TGAI , (EPA File Symbol No. 83623-G); a manufacturing use product (MP) , Hopguard L iquid (83623-R) ; and an end use product (E P ) , Hopguard II (EPA File Symbo l No. 83623-E). Simultaneous ly , M. Braverman (IR-4 Project) submitted a petition for an exemption from th e r equirement of tolerances for the active ingredient , Hops Beta Acids R es in (aka Potas sium Beta Resin , or KBR). In support of the re gistratio n and tolerance exemption petition , the registrant subm i tted product chemistry information for eac h product (MRJD 49251901 ) , field resi du e studies (MRJD 49251902) , sc ie nti fic rationales in suppo rt of the Tier I human heal th data requirements (MRJ D 49251904) , a guideline st udy for primary eye irritation (OCS PP 870. 2400 ; MRID 49251903), and sc ie ntific rational es in suppo rt of non-target organism data requirem e nt s (MRJD 49251905). Product chemistry studies and data have been reviewed under a separate Memorandum (See Memorandum from R. S. Jones toM. Adams , dated 09/ 03 / 2014). This portion of the review addresses the residue studi es , Tier I human health informati on , and Tier I non-target organisms informatio n . The EPA g ranted a Section 18 Emerge ncy Exe mption in 20 1 I in over 20 States to control Varroa mites (Varroa destructor) in honeybee h ives. The Sec ti on 18 was renewed in 20 1 2 and 20 1 3 in up to 37 States to control Varroa mites. It also is noted that in a literature review conducted in support of a GRAS determination by FDA (F DA , 20 01 ) , th e safety of all ext racted acids of hops (includ i ng Hops Beta Acids) is s upported by a l ong history of safe use in foods , es pecially beer , t hrou g hout r eco rded h is to ry . T h e literatur e review (FDA , 200 1) demonstrated that no information was available that was inconsistent with t h e safety determination for the proposed uses. EXECUTIVE SUMMARY Ther e are no concerns for human health w h e n P otass ium Sa lts of Hops Beta Acids (Hops Beta Acids) are used in accordance wit h Agency-app roved product use directions. In three years of field residue studie s , Hops Beta Acids were detected in only one sample of consumable honey (from the ho ney supers) at a leve l that was bare ly above the limit of detection (LO D 

### = 0.41 ppm); the re s idu es were present in the one sampl e at 0.44 ppm. T h e acu te oral toxicity LD50 = 1500 mg/kg (Chin and Ander so n , 1950) , a lev e l that i s approximately 3409X t he leve l found in one sample of honey. According to the National Honey Board (N HB , 2014) , annua l U.S. per capita honey consumption was approximately 1.3 I bs (approximate ly 0.6 kg or 600 mg) in 2013

. Assumi ng a 70 kg standard human and the lower acute oral toxic ity endpo int (LD so = 1500 mg!kg) , and a honey residue leve l of0.44 mg/kg , a human wou ld need to consume [( 1500 mg/kg x 70 kg)/ 0.44 mglkg) 

## = 2.39 x 10 6 kg] (approximately 5 lb s) of honey in one se r v in g to r eac h the acute tox icity endpoint of 1 5 00 mg/kg . The 5 lb level of hon ey consumption is approximate ly 3.9X greater than the annual U.S. per capita consumption of honey in 2013. It is noted here th at 2Hop Beta Acids PC Codes: 005 I I5 Review Type: Re sidue Studies , Tier I Tox, Tier I Non-targets DP Numbers: 4 I7758 & 4 I7759 EPA File No.: 83623-G, -R, & -E
the 0.44 ppm residue leve l for Hops Beta Acids was detected in o nl y one honey sampl e in three st

udies conducted between 20 10 and 2012. If ave raged ove r a ll honey samp l es, the residue level wou ld be far below the limits of detection. When fed to rats in drinking water at up to 50 ppm daily for 7 months, Hop s B e ta Acids have been shown to have potent anticarcinogenic activity against experi mentally-induced colon cancers with no observed toxic effects (Lamy et. al

. , 2007). Thi s subc hronic exposure in drinking water (50 ppm ) is approximately 114X g reater than the residues detected in one samp le of honey (0.44 ppm). There is a long history of oral/dietary exposure of humans to Hops Beta Acids via its u se as a preservative on meats (F D A, 200 1 ; USDA/FS IS, 2013) and its presence in beer making (Moir, 2000; Verze le 

### & De Keukelele , 1991 ), without any reported toxic effects. Hops Beta Acids are considered to be Generally Recognized As Safe (GRAS) by FDA (200 1 ). Hops B eta Acids have been s hown to inhibit inflammations in mice and exhib it anticarcinogenic activ ity without any known s ide effects (Cleemput et. al. 2009; Lamy et. al., 2007). Hops be ta ac id s are approved by the USDA/FSIS for use on read-to-eat meat products at up to 5.5 mglkg (ci t ed by Shen et. al. , 2009) to contro l pathog e n s, and has been approved for u se on control Listeria moncytogenes on frankfurters at up to 2.5 mg / pound on cooked meat and pou l try products (USDA/FSIS , 20 13). Acute, s ub chronic, and chronic exposure to Hop B eta Acids via inhalatio n and ocular routes of ex posure also is not expected and is highly unlikely. The TGAIIEP containing 16% Hops Beta Acids), is non-volatile , visco us , is not sprayed, and there is no dust associated with the strips so inhalation and ocular exposure is highly unlikely. Hand-to

- eye contact is mitigated by a label requi rement for eye protection. The most like l y expos ure to humans would r es ult from dermal ex posure to applicators whe n inserting the product- treated strips into th e bee hives. Dermal exposure will be mitigated by precautionary statements on the label requiring the use of gloves and eye protections w h e n app ly in g the product. Bee keepers already wear ex tensive PPE to protect themselves from bee stings, g l oves, p rotective suits, and a hat with a face mask or netting. There are no concerns for non-target organisms, including threatened and endangered species whe n the product is used in accordance with E PA- ap proved product labelin g. Based on a lack of expos ure , effects of Hops Beta Acids on non-targe t organis ms , including threatened and e ndangered species, are highl y unlikely and not ant icipated. Runo ff into g round and surface waters is not anticipated. The product (plastic strip s treated with a 16 % solution of Hops Beta Acids) is manuaJiy inserted into beehives. It is highly unlikely that organisms externa l to the hive wi ll have access to the treated strips, and li ttle or no likelihood that the active ingredient wi ll migrate from the hive to the external environment. In the highly unlike ly event that Hops Beta Acids would migrate out of the beehive to the ex te rnaJ environment, environmental fate data indicate that Hop s B eta Acids would degrade rapidl y, with 100% degradation in 36 hours in the light and 4 days in the dark (H uszcza, 2008). B ased on littl e or no likelih ood of exposure to any 

### 3Hop B eta Acids PC Co des: 0051 1 5 Review Ty pe : Residue Stud i e s, T ier I Tox , Tier I Non-targets DP Numbers : 417758 & 4 177 59 EPA Fil e No.: 83623-G , -R , & -E
non-target organis m , th e Agency makes a " No Effect " de terminati on for threatened and endangered spec ies.
RECOMMENDATIONS AND CONCLUSIONS 1. The petition fo r an exe mption of Potassium Salts of Hops Beta Acids from the req uire ment of tolerances is s upp o rted. 2a. Exce pt for Primary Eye Irritation , a s ub stantial amount of the information s ubmitted in support of th e data r e quire ment s for Tier I Human Hea lth Ass essme nt was not useabl e in that it did not s pec ifi ca ll y address the r espective routes of exposure. An Acceptabl e Prim ary Eye Irritat io n stu d y was submitted in s upp ort of the TGAI an d the E P. 2b. S uffi cient, accept abl e information (obtained from the t ec hnical li terature) as we ll as c re dible exposure rati onales were ava il ab le, how ever, t o support all of the T i er I Data require ments for both the TGAI and the EP. No additi o nal human health data are required at this tim e . 2c. Based on se lected acceptable in formation obtained fr o m th e technical literature, and/or acceptab le minimal ex posure rational es for each route of exposure for both the TGAJ and th e E P , no hum a n hea lth concerns are anticipate d when Hop s Beta Acids are use d in accordance w ith product label use directions. 3a

. Based on the method of application (manual in serti on of treated s trip s into the brood sup ers of bee hiv es) , there is littl e o r no lik e lih ood of any exposure to H op s Beta Ac id s to terrestri a l and aquatic non-target orga ni s ms. No additiona l non-target o rgan is m data are required at thi s tim e. 3b. B ased o n the lack of ex po s ure to no n-t arget organism s, a " No Effect '

. determinat ion i s m ade for threate ned and e ndan gered spec ies

STUDY SUMMARIES 

### Residue Data (40 CFR 158.2040) The reg is tr ant s ubmitted res idu e chemi stry dat a in support o f a request for an exe mption from th e require m e nt s of tolerances for Hops B eta Ac id s (PP 3E82 17 ). Residue chemistry data are not


### 4Hop Beta Acids PC Codes: 0051 15 Review Type: Residu e Studies , Tier I Tox , Tie r I Non-targets DP Numbers: 4 1 7758 & 417759 EPA File No.: 83623

- G , -R , &

- E typically required unless T ier II or Tier III toxicology studies are required and w he n numerical tolerances are required. Tier II or Tier III toxicology studies are not required for Hops Beta Acids and numerical tolerances are not requ ir ed for Hops Beta Acids at this time , although the registrant elected to submit burden of the residue studies and analytical methods for review. Three residue studi es were submitted in support of the r esidue data requirements

. The ftrst two studies were previously s ubmitted and reviewed in support of Section 18 Emergency Exemptio n requests ( 10ID0 6 , 100Rl2, and 10WA06) in 2010. Chemical Identity (OCSPP 880.1 00) : Hops Beta Acids (HBAs) are derived from the resin components of the cones of female hop plants. The three major components of Hop Beta acids are the structurall y-similar ofLupulone (30 -55 % with a isopropyl side chain) , Colupulone (20-55% with an isobutyl side chain); and Adlupulone (5-1 0% with a secbutyl s ide chain). See str uctures below. The components differ only in the R- side chain attached (see Table 2).

Lupulone Colupulone Adlupulone

Table 2

. Identity of Three Primary Hops Beta Acids Components Component CAS No. Mo l. Formu l a Mo l

. Weight R

- Side chain Lupulone (30 -55 %) 1 • 2 468-28-0 C26l-bs04 414.58 -CH2C H (CH3)2 Co lupul one (20-55%) 1 • 3 468-27-9 C2sl-b604 400.55

- CH(CHJh Adlupulone (5-10%) 1 • 4 31769-60-5 C21H3 s04 414.58

- CH(CH 3 )CH2CH3

.. 1 Percentages may vary accord in g to variety and growmg cond1t10ns 2 3,5-dihydroxy

- 2

- (3 -m ethy lbutan oy l}-4 , 6,6-tris(3 -methylbut-2-enyl)cyclohexa-2 , 4-dien-l-one 3 3 , 5-d ih ydroxy-4 , 6 , 6-tris(3-methylbut-2-eny 1)-2-(2-methylpropanoyl)cyc lohexa-2 , 4-dien- 1- one 4 3,5 -dih ydroxy-2,6,6-tris(3-me th yl-2-butenyl)-4-(2-methyl-1-oxobutyl)-2 ,4-cyclohexadien-l -on e 

### 5Hop Beta Acids PC Codes: 0051 15 Review Type: Re s idue Stud ies, Tier I Tox , Tier I Non-targets DP Numbers: 417758 & 417759 EPA Fi le No.: 83623-G, - R. & -E

The nominal concent rati on of Hops Beta Aci d s (Potassi um B eta Re s in , o r KBR) in the TGATi s 48.02% by weig h t of product , the remainder being co-extracted impuritie s (see Table 1) and intentionall y add ed inert ingredients [see Confide nti al Statement of For mul a, for HopGuard ® Technical Grade Ac ti ve Ingredient (Potassium Beta Resin= KBR), dated 10/21120 1 3; p 188 of 225 in MRID 49251901]. The nominal concentration of Hops B eta Acids respectively in th e M P [HopGua rd® Liquid ( Manufacturin g -u se Prod uct, MP)] and EP [HopGuard ® (E nd-u se Product)) is 1 6.01% and 16.0% (see CS F s da t ed 10/2 1/2 013 and 07 123/20 14). Deficiencies: None C l as s ific ation: ACCEPT ABLE

R es idu e Ana lyti ca l Method (OCSPP 860.1340) : The reg istrant s ubmi tted a me thod entitled , " QAM 05.14 Id e ntifi cation and assay for b e ta acids in honey and po ll e n " (See MRID 49 25 1 902 , pp . 28-48 for an ana ly tic a l s ummary , description of m ethod s, sampl e preparation , rep re se ntati ve chromatog ram s, standard curves, determination of minimum detectio n leve l s in hon ey (MDLs) , and cal cu l ations for limits of detection (LOD) and limit s of quantitation (L OQ) in honey). Th e ge neral scope of the met h od is to adsorb the be ta ac ids fo und in hone y and po ll en o nto a solidphase extracti on column and selectively de so rb the component s for analysis by HPL C. Th e method is based upon a published analytical method for hops beta acids (Ame rican Society of Brewing Chemists. M e thods of Analysis , 81 h Ed iti o n . Beer-23-C I so - a.-Aci d s by So lid Phase Extraction and HPLC ; & E urop ea n Br ewery Co nve nti on, Anal ytic a-Sec ti on 9 Beer Method). T he LO D for th e m e th o d is 0.4 1 ppm; the LOQ fo r the m ethod is 1. 2 ppm . Magnitude of the Residue in Honey (No O CS PP Number): In support of a request for an exem ption from the requirements of t olerances for potassium salts o f hop s beta acid s inio n hon ey (PP 3E8217), the r eg is trant submitted data from two field studies conducted in 2010 and a third in 2012. The first s tud y was conduc t ed during honey flow during late April/ May 2010; the sec ond study was conducted during honey flow dur ing late June /July 20 1 0; and the third study was conducted durin g nec tar flow from July 26

- August 23,2 01 2. In the fir st two s tudi es (2010), s in gle HopGuard app li ca tion s we r e use d. In the thi rd st ud y , three consec uti ve HopGuard a

pplications were u se d

. The fir st study was conducted at the be e ya rd of the US D AIA R S Carl Hayde n Bee R esearc h Ce nter (T uc son, AZ). S tudi es two and three were co ndu c t ed at the Casa Grande honey Bee Ap iary in (Tucson, AZ) by personnel fro m the USDA! ARS Carl Ha yde n B ee Research Ce nter. A n a lyses of residues in ho ney and pollen we re conducted at John I. Haas Laboratories ( Yakima , WA). Specific d e tail s of product application , honey collec ti on, sample 6Hop Beta Acids PC Cod e s: 005115 R e view Type: Residue Studie s, Tier I Tox , Tier I Non-targets DP Numbers: 417758 & 417759 EPA File No

. : 83623-G , -R, & -E
handling , storage conditions , and data for individual samples of each study are presented in MRID 49251902, pp. 49-66. The three studies are summarized below: 2010 ApriVMay Study: The first study was conducted in Arizona in commercial honey bee hives during late April and Early May 20 1 0. There were 8 colonies per trial w ith 6 treatment and 28 untreated control co l onies. Four treated co l onies had only one box with brood and the remaining two treated colonies had one honey box on top of the brood (bottom) box. HopGuard was app li ed to hives on cardboard strips (1.92 g a.i./strip). Two strips were hung over the brood frame in the bottom box only within eac h hive (3

. 84 g a.i ./ hive). After 35 days of exposure, honey samples were collected from each of the six top (honey) boxes and from the two bottom (brood) boxes. Honey samp l es were stored at -20 ° C until shi pment to at John I. Haas Laboratories (Yakima, W A) for analysis

. A ll adult bees and brood remained healthy and exhibited normal bee behavior throughout the 35- day exposure period. All strips were dry and had evidence of substantial chewing by bees. Residues were detected in 4 of 6 bottom (brood) boxes after 35 days exposure to HopGuardtreated cardboard strips. Residues ranged from 0.27 ppm to 9.52 ppm in 9 of 30 samp les from the brood boxes , and were not detectable in the remaining 21 samples. The average residue content of all 30 brood box samp les was 0.51 ppm. Hops beta acid residues in pollen obtained from treated frames contained an average of3.6 ppm over the 35-day exposure period

. Residue s were not detected in any samples obtained from the two top (honey) boxes (the ones used for human honey consumption). Residues were not detected in the untreated colonies. 2010 Jun e/July Study: The second study was conducted in Arizona in commercia l honey bee hives during late June and early July 2010. There were 20 co lonies per trial with two boxes of 10 frames each and a honey super. Ten colonies were treated with four HopGuard-treated strips (1.92 g a. i ./strip; two strips per box) in the bottom (brood) boxes for one week. Traps were monitored daily for dead bees during the one week exposure period and for 1 , 2 , 3 , and 7 days post treatment. Honey supers (not containin g brood), specifically de s igned for honey collection , were placed in the hive during nectar flow , and were removed from the hives after the one-week exposure period and set as id e for analysis of residues in th e honey. Honey samp les were stored at

- 20 ° C until shipment to at John I. Haas Laboratories (Yakima, WA) for analysis. According to the study auth or , 2010 was notable for an unseasonably l ow nectar flow , and only five treated colonies and one control co lony produced s ufficient honey for sampling. Except for the fust day (12 dead bees in treated hives , 6 dead bees in the controls) , numbers of dead bees were approximate l y the same in treated and control hives (6-8 dead bees per co l ony per day). The first day observations were considered to be not treatment related

. Residues of Hop Beta Acids were not detected in any honey samples obtained from the honey supers of 7Hop Beta Acids PC Codes: 0051 1 5 Review Type: Residue Studie s, Tier I Tox , Tier I Non-targets DP Numbers: 417758 & 417759 EPA File No.: 83623-G, -R, & -E

treated and untreated colon ies. Analysis of a spent strip at the end of the study , showed that after 35-days the HopGuard ® s trip contained 505 ug beta acids/ in2
, w hereas a fresh strip just out of the package contained 30,000 ug/i n 2

. 2012 July/ August Study: The third study was conducted in Arizona 

### in commercia l honey bee hives durin g late Jul y through late August 2012. There were 20 colo nies used in the study, each containing a simi lar number of frames of bees and brood , a queen, food storage and empty frames f or the queen to lay eggs. In mid-July , each colony was set up w ith honey supers. On July 26, two HopGuard ® stri ps (1.92 g a. i ./strip) were applied to each brood c hamber in 10 of the colonies. At one week post treatment , honey samp le s were collected from the honey s up ers, and the original HopGuard ® strips were replaced with new ones. At the end of the second week of treatme nt , a second honey sample was obtained from the honey supers of the treated colonie s. This process was repeated for a third week , for a total of three weekly HopGuard ® applications immed iate l y followed by weekly samp lin g of honey from the honey sup ers. Honey samp les were stored at -20 ° C until s hipment to at John I. Haas Laboratories (Yak ima, WA) for analysis. Colony activities (worker bee tasks of foraging , queen tending, brood rearing , cleaning , thermoregulation , guarding , comb building. etc.) , as well as queen egg laying were remained undisturbed by the treatments with HopGuard ® throughout the stu dy. H ops beta ac id s were not detectable in 21 of22 sampl es; in one sa mpl e, r es idue s were present 0.44 ppm which is barely above th e Limit of D etec tion (0.41 ppm) but below the Limit of Quantitation ( 1. 2 ppm). 

### CONCLUSIONS: When used in accordance with EPA-approved labeling , it is highly unlikely that the u se ofHopGuard® on the brood boxes of honey bee hives will result in detectable residues in edi ble honey. In three different studies conducted over three yea rs , only one honey samp le from one honey super was shown to ha ve residu es of hop s beta acids , and the residue leve l was only s lightly above the analytical limit s of dete c tion

. HopGuard ® is practically nontoxic to honey bees and was not observed to adversely affect normal hon eybee behavior. 

### Classification: ACCEPT ABLE

Review of Tier I Toxicity Data The regi s trant did not submit any Guideline stud ies 

### in support of the Tier I toxicity data requirements for the TGAIIEP. In lieu of Guideline studies , the registrant s ubmitted rationale s submitted by data and information from th e technical literature. These rationales are summari ze d below



### 8Hop Beta Acids PC Codes: 005115 Review T ype: Resid ue Stu die s, Tier I Tox, Tier I Non-targets DP N umbers: 417758 & 417759 E PA File No.: 83623-G , -R, & -E



### T ech ni ca l G rad e Active I n g red ien t/E n d-Use Pro d u ct (TGAI/EP) : Po t ass iu m Sa lts o f Ho p s Be ta Ac id s Reviewer's Note: The 

### TGAl is defmed as Potassium salts of Hops Beta Ac i ds and is present in the TGAl at 

### 16% (w/ w). The end

- use product consists of plastic strips coated with the 

### 16% Hop s Beta Acids solution and are contained in a sealed pouch until use. The plasti c strips are considered to b e the de l ivery system into the b ee hives (ab ov e the brood super) , and not an integral part of the formulation. The Agency co n s i ders the TGAl and the EP to be ide ntical for purposes of human health hazard and risk assessment. 

### A: Ac ute O ra l T ox icity ( OCSPP 87 0 .11 00) : The registrant did not submit Guideline s tudi es for either the TGA I or the EP, although none are requ ired at th is time. In l ieu of an acute ora l tox icity study, the registrant submitted a rationale supported by toxicity and ex posure information obtained from the technical literature (MRJD 49251904, pp . 5-7; and 27-29) i n fulfillment of the acute ora l toxicity data requirement for the TGAliEP ; the rele vant information i s presented below: 1. A s ing l e intragastric (oral gavage) administration of hops beta acids (as l upulon) to m i ce and rats resulted in LDs os of 1500 m g/kg and 1800 mg/kg , re s pectivel y (Chin and Anderson, 1950). These data classify Hops Beta Acids i n Toxicity Category III for acute oral toxicity

. 2. Hops beta acids are approved by the USDA/FSIS for u se on ready to eat meat products at up to 5.5 m g/ kg (cited b y Shen et. al., 2009) to control pathogen s, and has been approved for u se on control Listeria mon cytogenes on frankfurters at up to 2.5 m g/ products on cooked meat and poultry products at up to 2

. 5 m g/pound (USDA/FSIS , 2013). According to the NHDSC (2014) , US per capita consumption is approximately 70 hotdogs w ith an average weight of 

### 1.6 oz (45 g) per hot dog (Nutrition Facts, 20 14) or approximately 7 lbs of hot dogs per year. Fo r hot dog s alone , this would be equivalent to a d i etary intake of 17.5 mg Hop s Beta Acids/ person/year or an estimated daily dietary intake of0

. 048 m g/pe rson/day . 3

. In a review of information us e d to support FDA GRAS status , it was stated that there i s a long history of dietary exposure to Hops Beta Acids [comprised primaril y of lupulon e (C26H3&0 4), ad lupulone (C2s H 37 Q4), and colupu lone (C 2sH 3s0 4 )] , via beer consumption ( FDA , 2001 ). The same notice pro v ided estimates of dietary intake, via the us e of hops beta acids as antimicrobial agents in cooked ready-to-eat m eat s and on frankfurters , to b e up to 0

. 27 m g per pe rson per day

. A rev iew of the literature in support of the FDA GRAS status demonstrated that no estrogenic activi ty was observe d for polar extracts of hop s beta acids (FDA , 2001)

. 

### 9Hop Beta Acids PC Codes: 005115 Review Type: Re sidue Stud ies , Tier I Tox , Tier I Non-targets DP Numbers : 41775 8 & 417759 EPA Fil e No.: 83623-G, -R, & -E

4. The active in g red ient occurs naturally in the lupulin glands of th e flower s (cones) of hops (Humulus lupulus and contain up to 10% hop s beta acids (Moir , 2000; Verzele & De Keukelele, 1991 ). Hops are used extensively in the brewing of beer and have been used to inhibit acute localized inflammati ons in mice at concentrations of up to 0.4 ppm ( intravenous injection) with no side effects (Clee mput et. a l. 2009). 5. When fed to rats in drinking wate r at up to 50 ppm , hops beta acids have been shown to have potent anticarcinogen ic activity aga in st co lon cancers with no observed toxic effects (Lamy et. al., 2007). 

### Conclusions: Hop s B eta Acids can be classified in Toxicity Category III fo r acute oral toxicity, based on studies where mice and rats were dosed w ith Hop s Beta Ac id s by oral gavage; Observed LD50 s ranged between 1500 to 1800 mg/ kg (Chin and Anderson , 1950). ln addition, acute oral expos ure to Hops Beta Acids is highly unlikely. H ops Beta Acids are applied inside the hive to the brood supers, on p lastic strip s treated with a 16 % s olution of Hop s Beta Ac id s (two strips per hive; 1.92 g/s trip). Residues have been shown not to exceed 0.44 ppm (detected in only o ne sa mple of ed ibl e honey in three years of stud ies; see re s idues studies above), well be low any known endpoints fo r Hops B eta Ac id s ( 1 500 mg/ k g). No additional data are required at this tim e. There is a long hi story of oral/ dietary exposure of humans to Hop s Beta Acids via its use as a preservative on meats (FDA , 2001; USDA/FSIS , 20 13) and its presence in beer making (Mo ir , 2000; Verzele & De Ke uke lele , 1991 ). Hop s Beta Acids are considered to be Generally Recognized As Safe (GRAS) by FDA (200 I ). Hops Beta Acids have bee n shown to inhibit inflammations in mi ce and ex hibit anticarcinogenic acti v it y without any known side effects (Cleemput et. a l. 2009; Lamy et. a!., 2007). 

### Classification: ACCEPTABLE

B: Acute Dermal Toxicitv (OCSPP 870.1200): The registrant did not submit Guideline studies for either the TGAI or the 

### EP, althoug h none are required at th is tim e. Dermal exposure to the active ingredient is highly unlikely based on the method of app li cation (manual appl ication of treated plastic strips into beehives) , and the requirement of worker/app li cato rs to wear gloves when applying the end-use product. Non-occupational exposure to th e active ingred ien t doe s not occur.

10Hop Beta Acids PC Codes: 0051 15 R

eview Type: Residue Studie s, Tier I Tox , Tier I Non-targets DP Number s: 417758 & 417759 EPA Fil e No. : 83623-G , -R, & -E
Ho weve r, in li eu of an ac ute dermal toxicity stud y , the reg istrant s ubmitted a rationale supported by toxicity and exposure information (MRID 49251904 , p. 8; and 30-31) obta in ed from the technical literature in fu lfillment of the acute dermal toxicity data requirem e nt for the TGAI. Much of the information was not useable in that it did not specifically address the acute dermal toxicity rou te of exposure. One study (Dumas , et. al. , 2009) reported the effects of a deodorant st

udy whereby 0.4 g of a mixture ofHops Alpha Ac id s (50%), Hops Beta Ac id s (26%), and other s ubstances (zinc , ricinoleate) present at unknown concentration s , were applied to human underarms as a deodorant stick. Preliminary studi es demonstrated that the ex tract inhibited the growth ofunderann odor cau si ng bacteria in Petri dish cultures

. Each sub j ect received two app li catio ns of the deodorant s tick , although th e precise amount ofl·lops Beta Acids applied is uncertain

. D erma l irritation and toxicity were not observed in any test subject. Due to the mixture of substances u se d as the test substance , thi s study cannot be u se d to fulfill the data req uirement for acute derma l toxicity. 

### REV IEWE R 'S NOTE and Co nclu s ion s: The sub mitted information (MR JD 49251904 , pp. 8; and 30-31; and Dwna s, et. a l. , 2009) did not spec ifi cally address the acute dermal toxicity route of exposure. In add i tion , humans were used as th e te s t s ubj ects a nd the study was not submitt ed to the Hum an Studies Review Board (HSRB) fo r a review of its suitabi lit y for u se in s upporting a pestic ide registration. The BPB reviewer notes that acute dermal exposure to Hops Beta Acids is highly unlikel y and would only occur to the worke r / applicato r. Based on the method of application ( manually applied v ia plastic strip s in s ide beehives ), non- occupational exposure to Hops Beta Acids wi ll not occur . Occupational ex posure is mitigated by protective clothing nonnally worn by beekeepers, including gloves , protective su it s , and a hat with a face mask or netting. Precautionary statements o n the labe l must require the use of g lo ves and eye protection for workers/ app li cators. 

### C la ss ification : ACCEPTABLE , based on a lack of acute exposure via the dermal route of ex po s ure. It is recomm e nded that precautionary s tatements requiring the u se of gloves and eye protection for workers / applicators be placed on th e product label , based on the results of the Primary Eye Irrit ation s tud y (see below)

.


### C: Acute Inhalation Toxicitv (OCSPP 870.1300): The reg istrant did not s ubmit Guideline stud ies for e ither the TGAI or the EP, a lthough non e are requ ired at this time. Inhalation ex posure to the active ingredient is highly unlikely based on the non-volatile nature of the active ingredient and method of application (manual application of treated plastic s trips into beehives. Non-occupational exposure t o the active ingredi e nt does not occur. Th e regis trant also provided the following exposure rationale: '·HopGuard (the end-use product contain ing 16% Hop s Beta IIHop Beta Acids PC Codes: 005 1 15 Review Type: Res id ue Studies , Tier I Tox , Tier I Non-tar gets DP Numb er s : 4 1 775 8 & 4 17759 E P A File No.: 83623 -G , -R, & -E
Acids), is non-volatile, viscous, is not s pra yed, and th e re is no du s t associated with the strips so inhalation exposure is highly unlikely. "
Th e reg i strant a l so s ubmitte d toxicity and exposure in formation (MRIO 4925 1 904, p. 9; and 32

- 33 ) obtained from the tec hnj callit e rature in fulfillment of the acute inha lation toxicity data
req uirement fo r the TGA I / E P

. However, much o f th e information was not u seable in that it did not spec i fica ll y a ddress the ac ute inhal a ti on toxic ity ro ute of exposure. 

### Con clu s ion s: Du e to the lack of vo latility a nd th e m e thod of product application (manual in sertion of Hops Beta Acids-treated s trip s into beehjv es), occupational and non -occ upati o n a l inh a latio n exposure is hl g hl y unlikel y. No additional data are required at this tim e. 

### C la ss ification: ACCEPTABLE , b ase d on a lack expos ure via th e inhalation rout e of exposure



### D: Primary Eve Irritation (O CSPP 870.2400): A Guideline prim ary eye irritation study (M RID 4925190 3) was conducted u s in g p otass ium salt s of Hop s B eta A cid s ( the TGAI) o n rabbit s. The n o rmal pH of the test sub s tan ce is pH II and was a dju s ted to pH 9 prio r to u se. Eac h of three te s t rabbit s was administered a 0.1 mL of the test s ub s tance to th e conjunctiva l sac of the right eye ; the left eyes we re u sed as the controls. Test and contro l eyes were observed f o r sig n s o f irritati o n fo r 2 1 d ays post treatment. Corn ea l opac ity , iriti s , and conjunctivitis were observed a 1-hour post treatment. By D ay 2 1 po st treatme nt , non e of the ocular res ponse s had complete ly r eso l ved in any o f the test eyes. 

### Co nclu s ion s: Based on the data , Potass ium Salt s of H o p s B eta Acids are cl assified as Co rrosive , Toxicity Cate gory I. 

### C lassificatio n : ACCEPT ABLE

E: Primary Dermal Irritati o n (O CSPP 870.2500): The registran t did no t s ubm it Guidelin e s tud ies for e ith e r the TG A f or th e E P , although non e a r e re quired a t thi s time

. D e rmal ex po s ure to the active ingredient is hi g hl y unlike l y based on the method of app l ication ( manual application of treated plas ti c stri ps into beehives) , and the requirement of worker / a pplic ators to wear g l oves when app l yi n g th e end

- u se product. Non-occupationa l ex posure to the active in gre dient do es not occur. H owever , In li eu of a primary dermal i rritation study , the registrant s ubmitted a rationale
s upp orted b y to x icity and ex po s ure inf o rm a tion (MRID 49251904 , pp. 11-1 2; and 35 -36 ) obtained from th e tec hn ica l lite rature in fulfillment of th e prim a r y dermal irritation data 1 2Hop Beta Acids PC Codes: 005115 Review Type: Residue Studies, Tier I Tox , Tier I Non-targets DPNumbers: 417758&417759 EPA Fi le No.: 83623-G , -R, & -E requirement for the TGAl!EP. Much of the inform at ion was not u seable in that it did not specificall y address the primary dermal irritation route of exposure. The registrant also provided the following exposure rationale: 

### "Contact with growing hop plants is known to cause an allergic contact dermatitis and is attributed to the myrcene oil in .fresh hop oil. Growing hop plants can also cause a mechanical dermatosis attributed to the rough climbing hairs on the stem and s e cretions of the yellow glandular hairs on hops. It is hypothesized that the hop maturingprocess may turn certain hop proteins into allergens, I rare cases leading to a diagnosis of immunological contact urticaria. Hops Beta Acids were not implicated in this process (Estrada et. a/., 2002).
In addition, one study (Dumas, et. a l. , 2009) reported the effects of a deodorant study whereby 0.4 g of a mixture of Hops Alpha Ac id s (50%), Hops Beta Acids (26%), and other s ub stances (zinc , ricinoleate) present at unknown concentrations, were applied to human underarms as a deodorant stick. Each subject r eceived two applications of the deodorant stick , although the precise amount of Hops Beta Acids applied is unc ertai n. Dermal itTitation and toxicity were n ot observed in any test subject. Due to the mixture of substances used as the test s ub stance, this study cannot be used to fulfill the data requirement for primary dermal irritation , although it does provide s uppl emental information. 

### REVIEWER'S NOTE and Conclusions: The submitted information (MRI D 49251904, pp. 11-12; and 35-36 ; Dumas , et. a l. , 2009; and Estrada et. al., 2002) did not spec ificall y address the dermal irri tat ion route of exposure. In addition , humans we re u sed as the test s ubj ec ts in both cited studies and they were not submi tted to the Human Studies Review Board (HSRB) for a review of its suitability for use in support in g a pesticide registration. The BPB rev iewer notes that dermal exposure to Hops Beta Ac id s is highly unlikely and would o nl y occur to the worker / app li cator. Based on the method of application (applied via plastic strips inside beehives) , non-occupational exposure to Hops Beta Acids will not occur. Occupational exposure is mitigated by protective clothing normally worn by beekeepers, includ in g gloves , protective suits , and a hat with a face mask or netting. 

### Classification: ACCEPTABLE, based on the rationale (Estrada et. a l. , 2002) subm itted above and the mitigation of expos ure with appropriate PPE. It is recommended that Tox ici ty Category I PPE precautionary lan g ua ge for ac ute dermal toxicity be placed on the label , based on the results of the Primary Eye Irri tation study (see above).



### E: Dermal Sensitization (OCSPP 870.2600): The registrant did not s ubmit Guideline studies for either the TGAI or the EP , although none are required at this time. Dermal exposure to the active ingredient is highly unlikely based on the method of application (manual application of 13Hop Beta Ac id s PC Codes: 005115 Review Type: Residue Studie s , Tier I Tox , Tier I Non-targets DP Numbers: 4 17758 & 417759 E

PA File No.: 83623-G , -R, & -E

treated plastic strips into bee hi ves), and the requirem ent of worker /applicators to wear gloves when app lyi n g the end-use product. Non-occupat ional exposure to the active ingredient d oes not occur. However, in lieu of a dermal se n s iti zation study, the r eg istrant s ubmitted a r a ti ona l e s upported by toxicity and exposure information (MRlD 49251904, p. 1 3; and 37 -38 ) obtained fro m th e technical literature 

### in fulfillm ent of the dermal se n s iti zation d ata requirement fo r the TGAl/EP. Much of th e information was not u seab le in that it did not spec ifi cally address primary de rm al sensitization. Howe ve r, th e registrant did prov ide the following ex posur e rati o nal e: "Con ta ct with growing hop plants is known to cause an allergic contac t dermatitis and is attributed to the myrcene oil in fresh hop oil. Grow ing hop plants can also cause a mechanical dermatosis altributed to the rough climbing hairs on the stem and secretions of the yellow glandular hairs on hops. It is hypothesized that the hop maturingprocess may turn certain hop pro teins into allergens, I rare ca ses leading to a diagnosis of immunological contact urticaria. Hops Beta Ac id s were not implicated in this process (Es trada e t
. a!., 2002). Sim ilar to the conclusions reached for primary dermal irritation (see above) , the BPB re viewe r notes that dermal exposure to Hops Beta Acids is highly unlikely and wo ul d only occurto th e worker /appl icato r. B ased on the method of applicat ion (applied via plastic strips inside beehives) , non- occupational exposure to Hops Beta Acids will no t occur. Occupational exposure is miti gated by protectiv e clothing normally worn by beekeep e rs, includin g g l oves, protectiv e s ui ts, a h at with a face ma sk or nettin g . Precautionary stat eme nt s on the labe l mu st require the use of g loves and eye protection for workers /app licators. 

### Classification: ACCEPT ABLE , based on the rationale (Estrada et. al. , 2002) submitted above and the mi ti gation of exposu re with appropriate PPE. It is recommended that Tox icity Category 1 PPE pr ecautionary lan g ua ge for ac ute de rmal toxicity be place d on the labe l, b ased on the re s ult s of the Pri mary Eye l rri tation stu dy (see above).



### F: 90-Day Oral Toxicity (OCSPP 870.3100): T he registrant did not sub mit Guideline studies for e ithe r th e TGA I or the EP , although no ne are required at thi s time. Rep eated , s ubchroni c exposure to the active in g r e dient i s highl y un lik e ly bas e d on the method o f app li cation (manual application of treated plastic stri ps into bee hi ves). Di etary ex pos ur e also, is hi g hl y unlikel y, based on the lack of residues in e di b le honey (see residue stu di es above). In three different studies conducted over three years , o nl y one honey sample from one honey super was shown to have residues of hop s beta acids (0.44 ppm) , and the r es idu e level was only slightly above the analytica l limit s of d etec tion (0.41 ppm) and well below the Limit of Quantitat ion ( 1.2 ppm).

14Hop Beta Acids PC Codes: 005 II 5 Review T y pe: Residu e Studi es, T ier I Tox , T ier I Non-targets DP Numbers: 417758 & 417759 EPA File No

. : 83623-G, -R. & -E

In addition , the r egistrant submitted a rationale supported by toxicity and exposure information obtain ed fi·om the technical literature (MRJD 49251904, pp. 14-16; and 39-41) in fulfillment of the 90-day oral toxicity data requirement for the TG AIIEP. The relevant information, much of which i s s imil ar to that presented in suppo rt of the acute oral toxicity data requirement, is presented below: 1. Hops beta acids are app ro ved by the USDNFSIS for u se on ready to eat meat products at up to 5.5 mglkg (c ited by Shen et. al. , 2009) to contro l pathogens , and has been approved for use on control Lis teria moncytogenes o n frankfurters at up to 2.5 mg/pound on cooked meat and poultry products (USDAJFS IS , 2013)

. 2. In a review of inf ormation used to support FDA GRAS sta tu s, it was stated that there i s a lo ng history of dietary exposure to Hops Beta Ac id s [compri sed primarily of lupulone (C2 6HJs04) , adlupulone (C2s H 3 7Q4 ) , and colupulone (C2 sH 3 s04) ] , via bee r cons umption (FDA , 2001 ). The same notice provided estimate s of dietary intake, via the use of hops beta acids as antimicrob ial agents in coo ked ready-to-eat meats and on frankfurters, to be up to 0.27 mg per person per day. A review of the literature in support of the FDA GRAS status demonstrated that no estrogenic activity was observed for polarity extracts of hops beta acids (F D A , 200 1). 3

. The active in gredient occurs naturally in the lupulin glands ofthe flowers (cones) of hops (Humulus lupulus and contain up to 10% hops beta acids (Moi r , 2000; Verzele & De Keukelele , 1 991 )

. Hops are used extensively in the brewing of beer and have been used to inhibit acute localized inflammations in mice at concentrations of up to 0.4 ppm (i ntra venous injection) w ith no s id e effects (C l eemp ut et. al. 2009). 5. When fed to rats in drinking water at up to 50 ppm daily for 7 months , hops beta acids have been shown to have potent anticarcinogenic activity against experi me nta ll y-induced colon cancers wi th no observed toxic effects (Lamy et. al., 2007). 

### Conclusions: T h ere is a l ong hi story of oraVd ietary exposure of humans to Hops Beta Acids v i a its u se as a preservative on meats (F D A , 2001 ; USDA!FS IS, 2013) and its presence in beer making (Mo ir , 2000; Verzele & De Keukelele , 1991) , without any reported tox i c effects. Hops Beta Ac id s are cons id ered to be Generally R ecognized As Safe (GRAS) by FDA (2001). H ops Beta Acids have been shown to inhibit inflammati ons in mice and exhib it anticarcinoge nic activity without any known side effec ts (Clee mput et. al. 2009; Lamy e t. al. , 2007). It is also noted th at rat s have been administered Hops Beta Acids in drinking water at levels up to 50 ppm with no observed toxic effects. Tllis subchro nic exposure in drinking water (50 ppm) i s Approximately 114X greater than the residues detected in one sample of ho n ey (see residue 1 5Hop B eta Acids PC Codes: 005115 Rev iew Type: Residue Stud ies , T ier I Tox , T ier I Non-targets

s tud ies above). C l ass i ficat i o n : ACCEPT A B LE DP Numbe rs: 417758 & 417759 EPA Fi le No.: 83623-G , -R , &

- E

G: 9 0

- D av D erm a l Tox icity ( O CS PP 87 0 .325 0 ): The registran t did not s ubmit Guideline s tudies for either th e TGA I o r the EP , although none are required at thi s time. Repeated dermal exposur e to the active in g r ed ient is hi ghl y un lik e l y based on the method of ap pli cation (m anual application of treate d plastic stri ps into be e hives), and the requ irem e nt of worker / applicators to wear g loves when applying the end-use product. No n-o cc upational ex po s ure to the active ingredient doe s not occu r. Th e registrant s ubmitted a rationale supported by toxicity and exposure inf ormat io n obtained from the t echni cal literature ( MRID 49251904 , p . 16 ; and 42

- 43) in fulfillment oft he 90-day dermal toxicity data requirement for the TGAI/E P

. Much of the information was not u seable in that it did not s pec i fically address subchronic ( 90-day) dermal toxicity. However , the registrant did provide th e fo ll owing expos ure ratio na le : " Contact with growing hop plants is known to cause an alle r g i c con ta ct dermatitis and is allributed to the rnyrce ne oil in .fresh hop oil. Grow in g hop plant s can also ca use a m echa nical dermatos is allributed to the rough climbing hairs on the stem and secre tions of the yellow g landular hairs on hops. It is hypothesized that the hop maturing pro cess ma y turn ce rt ain hop proteins into allergens, I rare cases leading to a diagnos is of immun ological contact ur ticaria. H ops Bet a Acids we re not im plicated in this p r ocess (Es trada e t
. a!. , 2002). REVIEWE R 'S NO T E and Co nclu s i o n s: The submitte d informati o n ( MRlD 49251904 , p . 1 6; and 42-43 ; and Estrada e t. al. , 2002) d id no t specifica ll y address sub c hron ic derma l tox icity. However, the BPB r ev iewer notes that rep eated , subchronic exposure to Hop s Beta Acids i s hi g hly unlikely b ased o n th e me thod of appl ication (manually in serted into beehives), low app lic ation rate , lack of appreciable residues in ho ney , and appro priate PP E (g loves and eye pro tection) worn by workers/ app licators. C l ass i fi c a ti o n : ACCEPT ABLE , based on minimal exposure and a ru story of exposure with no observed to xic i ty.

H: Ac ute Inh a l a ti o n Tox i c ity (O CS P P 87 0 .3 4 65 ) : The regi s trant did not s ubmit Guideline s tudies for e ith er th e TGA I or the EP , although no n e are r eq uired at thi s time. Repeated , s ubchronic inhalati on expos ur e to the active ingredient is ru ghl y unlike ly ba se d o n the nonvo latile nature of the active ingredient and me thod of application (manua l application of treated plas ti c strips into beehi ves. No n- occupatio n al exposure to the active ingredi e nt does n ot occur. 16Hop B eta Acids PC Codes: 005115 Review Type: Residu e Studi es, Tier I Tox , Tier I Non

-targets DP Numbers: 417758 & 417759 EPA Fi le No.: 83623-G , -R, & -E

The registrant also provided the following exposure rationale: "HopGua rd (the end-use product contain ing 16% Hops Beta Acids) , is non-volatile, viscous , is not sprayed, and th ere is no dust associated with the strips so inhalation exposure is highly unlikely. " The registrant also submitted toxic i ty and expos ure information (MRJD 49251904, p. 17 ; and 44- 45) obtained from the technical literature in fulfillment of the acute inh alation toxicity data requirement for the TGAIIEP. However , much of the infom1ation was not useable 

### in that it did not specifically address the acute inhalation toxicity route of exposure. Conclusions: Due to the lack of vo latility and the method of product application (manual ins ertion of Hops Beta Acids-treated strip s into beehives) , repeated, s ubchronic occupational and non-occupational inhalation exposure is highl y unlikel y. No additional data are required at this time. Classification: ACCEPTABLE , based on a lack exposure via the inhalation route of exposure.

1: Pre-Natal Development-Rat (OCSPP 870.3700): The registrant did not submit Guideline studies for either the TGAI or the EP, although none are required at this time. Use of the product under widespread and commonly recognized practice will not result in significant exposure to female humans. Occupational and non-occupational exposure is no t expected due to the method of application and the lack of appreciable residue s in honey (see residue studies above)

. Hops Beta Acids are applied inside the hiv e to the brood s upers , on plastic strips treated with a 16 % so lution of Hop s Beta Acids (two strips per hive; 

### 1.92 g/strip). Residues have been shown not to exceed 0.44 ppm in honey (see r es idu es studies above) , well below any known acute (1500 mg/kg ; Chin and Anderson , 1950) or s ubchronic (50 ppm in drinkin g water; Lamy et. al. , 2007) endpoints for Hops Beta Acids. No additional data are req uired at this tim e. The registrant also submitted toxicity and exposure information (MRID 49251904 , p . 18-19; and 45-47) obtained from the techn i cal lite rature in fulfillment ofthe acute inl1alation toxicity data requirement for the TGAIIEP. Ho weve r , much of the inform ation was not useable in that it did not specifical l y address pre-natal d evelop ment. Co nclusion s: Based on the method of product application (manual in sertion of Hops Beta Acids-treated str ip s into beehives) , a lack of acute or subchronic exposure , lack of significant exposure to female humans , and lack of appreciable residues in edible honey (see residue studies above) , pre-natal developmental toxicity is h ighly unlikely. No additional data are re quired at this time. C l ass ific ation: ACCEPTABLE 17Hop B eta Acids PC Codes: 0051 15 Review Type: Re s idu e Srudi es, T ier I T ox , Ti e r I Non-tar gets DP Num bers : 417758 & 4 1 7759 E PA F il e No.: 83623 -G , -R, & -E

1: Mutagenicity Testing (OCSPP 870.5100,870.5300 , & 870.5375): The reg i strant did not submit Guideli ne studies for e ither the TGAl or the EP , although none are r eq uired at thi s time. Occupational and non-occupational exposure is not expecte d due to the me thod of application and th e lac k of appreciable r es idues in hone y (see residue st udi es above). Hop s Beta Acids are app lied in s ide the hi ve to the brood s up ers, on p las tic strips treated with a 16 % so lution of Hops Beta Aci d s (two strips per hive ; 1.92 g/stri p). Re si due s ha ve bee n s ho wn not to exceed 0 .44 ppm in hon ey (see re s idue s stud ies above) , we ll below an y known ac ute ( 1500 m g/kg; Chin and Ander so n , 1950) or s ubchronic (50 ppm in drinking water; Larny et. al. , 2007) e ndpoints for Hop s B e ta Ac id s. No additional data are required at this tim e . The registrant also subm itted toxicity and ex posur e informati o n ( MRID 49251904 , p . 18-19 ; and 48-54) o bta ined from the technical literature in fulfillment of th e mutag e ni c ity data requirements for th e TGAI/E P. How eve r, much of the information was no t u seable in th at it did not specificall y address mutagenicity. Conclusions: Based on the m ethod of product application (manual insertion of H ops Beta Ac ids-treated strips into beehives) , lack of acute o r subchronic ex po sur e , long history of exposure to H ops B e ta Ac id s 

### in food preservation and in th e makin g ofbeer , lack o f appreciable residu es in edible honey , and the lack of any s tructural or chemical similarity to known mutage n s , mutage ni c i ty is hi ghly unlike ly. No ociditional data are r eq uired at thi s time. Classification: ACCEPTABLE
QUALITATIVE HUMAN HEALTH RISK ASSESSMENT
Acute , s ub c hronic , and c hro nic exposure to Ho p Beta Acids v ia the oral route of ex posur e is not expected and is highly unlike ly. Residu e d ata s ubmitted by th e applican t indi cate th at re s idue s of H op Beta Ac id s w ill lik ely not be pr esent in ed ible hon ey , when only the brood boxes (supers) of a hon ey bee colo ny/hive are treated with th e e nd-u se produ ct (a pla stic strip coate d with 1.9 2 g a.i with two s trip s/b rood b ox)

. Brood box hon ey i s not us e d for human con s umption

. A series of thre e fie ld studies conducted be tween 201 0 and 20 12 demon strated that w he n the product is app li e d according to la be l u se direction s , h o ney obtained from the hon ey boxes (s upers) , hon ey u se d for human consumption , was virtually residue free. O nl y o ne s ampl e of ho ney (o ut o f the three field s tu dies) h ad residues of H op s B eta Aci d s pre se nt at 0.44 ppm , barely above th e Limit of D e tect io n of (0.4 1 ppm) and s ub stantially be low the Limit o fQuantitation ( 1 .2 ppm). Mea s ure d acute oral to x icity e ndpoint s (LD sos) for Hop s Beta Acids rang e from 1500 mg!kg to 1800 m g/kg in mic e and rat s by oral gavage (C hin and And ers on , 1950). Th e lower acute ora l toxicity e ndpoint i s 3409X g reater than th e s in g l e me as ured r es idue leve l in ho ney (0.44 ppm ). A 18Hop Beta Acids PC Codes: 0051 15 Review Type: Residue Studies, Tier I Tox , Tier I Non-targets DP Numbers: 417758 & 417759 EPA File No.: 83623-G , -R, & -E

subchron i c study where in rats were administered Hops Beta Acids at 50 ppm daily for 7 months in drinking water reported no toxic effec ts (Lamy et. al. , 2007)

. The same study also observed substantial anti

- carcinogenic effects in rats that were chemically-induced to have colon tumors. According to the National Ho ney Board (NHB, 2014) , annual U.S. per capita honey consumption was approximate l y 1. 3 lbs (app rox im ately 0.6 kg or 600 mg) in 2013. Assuming a 70 kg standard human and the lower 

### LDs o acute oral toxicity endpoint of 1500 mg/kg , and a honey residue leve l of 0.44 mglkg , a human would need to consume ((1500 mg/kg x 70 kg) / 0.44 mg/kg) 

### = 2.39 x I 0 6 kg (approximately 5 lbs) of honey to reach the acute toxicity endpoint of 1500 mg/kg. The 5 lb level of honey consumpti on is approx im ate ly 3.9X g reater than the U.S. per capita consumption of honey in 2013

. It is noted here that the 0.44 ppm residue level for Hops Beta Acids was detected in only one honey sample in three studies conducted between 20 I 0 and 2012. If averaged over all honey samples , the residue level would be far below the limits of detection. Acute, subchronic , and chronic exposure to Hop Beta Acids via inhalation and ocu lar routes of exposure also is not expected and i s highly unlikely. The TGAI/EP containing 16% Hops Beta Acids) , is non-volatile , v iscous , is not sprayed , and there is no dust associated w ith the strips so inhalatio n and ocular exposure is highly unlikely. Hand-to-eye contact is mitigated by a label requirement for eye protection. The most likely exposure to humans would result from dermal exposure to applicators when inserting the p roduct -t reated strips into the bee hives. Dermal exposure will be mitigated by precautionary statements on the label requiring the use of gloves and eye protections when applying the product. Bee keepers already wear extens ive PPE to protect themselves from bee stings, g loves, p rotective suits , and a hat with a face mask or netting. CONCLUSIONS: When the product is used according to l abel use directions , there are no human health concerns w ith the use of Hop Beta Acids in bee hives to control Varroa mites at this time.

Review of Tier I Non- Target Organism and Environmental Fate Data The registrant did not submit any Guideline studies in support of the Tier I toxicity data req uirements for the TGAI/EP. In lieu of Guide line studies, the registrant submitted rationales submitted by data and information from the technical literature. These rationales are summarized below

19Hop Beta Acids PC Codes: 005 I I 5 Review Type: Residue S tud ies , Tier l Tox , Tier I Non-targets DP Numbers: 4 I 7758 & 4 I7759 EPA File No.: 83623

- G , -R, & -E
Technical Grade Active Ingredient/End-Us c Product (TGAI/EP): Potassium Sa lts of Hops Beta Acids Reviewer's Note: Th e T GAI is defined as Pot assium salts of Hops B eta Acids and is present in the TGAI at 16 % (w / w). The end

- use product consists of pl astic strips coated with the 16% Hops Beta Acids sol uti on and are contained in a sealed pouch until use. The plastic strips are considered to be the de li very system into th e beehives (above the brood super) , and not an integral part of the formulation. The Agency considers the TGAI and the E P to be identical for purposes of non-tar ge t organism hazard and ri sk assessment. Overall, the rationales are based on a lack of expos ure to the active ingredient due to the manner in which the end-use product is applied to co ntro l Varroa mites in beehives. Hop Beta Acidstreated are applied to the inside of honey bee hives (colonies) on product-treated cardboard strips. The two strips eac h (four per hive) are pl aced on brood frames (bottom boxes of the commercial bee hive). T hi s method of application, as we ll as the low application rate (1.92 g a.i ./ cardboard strip), preclude s any sig nificant exposure to any non-target organisms or the environment external to the bee hive. In th e highly unlikely eve nt that Hops Beta Acids would migrate out of the beehive t o the ex ternal environment, environmental fate data indicat e that Hops Beta Acids wou ld degrade rapidl y, with I 00% degradation in 36 hours in the li g ht and 4 days in the dark (Huszcza, 2008). Avian Acute Oral & Av ian Dietary Toxicitv (OCSPP 850.2100 & 850.2200): It is highly unl ikely that there will be any ac ute oral or dietary exposure of hop s beta ac ids to birds via the application ofHopGuard ® (16.0% a.i.) to the ins ides of commercial beehives. There is little or no opportunity for a bird to gain access to the active in g redient-treated cardboard strips inside a beehive, nor i s there a li keli hood that the act ive in gred ien t will mi g rate outside of the hive after app li cat ion . In the unlikely event that hop s beta acids would migrate outside the beehive , they would be rapidly degraded by soil fungi. In laboratory experiments , Trametes versicolor was shown to fully degrade humulon es and lupulones in commercial hop extract and sterilized spent hops after 4 days incubati on in th e dark and 36 hours in the light (Hu szcza, 2008). In add ition , limited testing data indicate that chic ken s administered up to 250 ppm of a hops beta acid ( lupul one) in drink in g water for 1 3 to 22 days post hatchin g had no effects on bird behavior or weight gain (Siragusa et. a !. , 2008), and improved th e ir health by inhibiting pathog e nic clostrid ia in the ch i cken intestina l tract. In experi m ents desi gned to evaluate the effects of brewery waste in the di e ts of Muscovy ducks , it was observed that brewery waste (known to contain hops beta acids) could replace up to 50% of the diet without reducing daily weight ga in s (Dong and Og le, 2003). The Agency has granted Section 18 Emergenc y Exemptions in 20 I I in over 20 States to con trol 20Hop B eta Acids PC Codes: 0051 1 5 Rev iew Type: Residue Stud ies , Ti e r I To x , Ti er I No n-targe ts DP Numbers: 417758 & 417759 EPA File No.: 83623-G , -R , & -E

Varroa mites (Va.rro a d estructor) in hon ey b ee hi ves, whjc h we r e renewe d in 20 12 and 2013 in up to 37 State s to co ntrol Va rroa mite s. No adve r se effec t s have ever been r e p o rted for bird s. C onclusion: B ased upon a lack of exposure t o birds v ia the use of th e e nd-u se p roduct in bee hi ves, no adverse effec t s are expected in bird s a s a result of the u se o f HopGuard® ( 16.0 % potass ium salts of hop s beta acids). No toxici ty is expec ted in the unlikel y eve nt that birds are orall y exposed to h op s beta ac id s . The r e are no co ncern s for non-targe t birds. C la ss ification: ACCEPTABLE

Freshwater Fish Acute Toxicity & Freshwater Aquatic Invertebrate Toxicity (OCSPP 850.1085 & 850.1010): It is hi g hl y unlike ly that th e r e w ill be any exposure of potassium salts of hops beta aci d s to aquat ic o rgan is m s v ia the ap pli cation o f H opGuard ® ( 16.0% a.i.) to the in s id es of com me rc ia l beehives

. There is li ttle or no opportunity for an aquatic organism to ga in access to the active in gr e di e nt-trea ted cardboard stri ps in s id e a beehiv e, nor is there a likelihood that the act ive in g r edient w ill mi g rate outside of th e hi ve and into aquatic e nv iro nments after a ppli cation.
In the unlikely even t that hops beta acids wo ul d mi g r ate o ut s id e the beehiv e , th ey would b e rapidly deg raded by soi l fungi. In laboratory exper ime nts , Trameles ve r sico lor was s hown to fu ll y d eg rade humul o nes and lupulones in commerc ia l hop extract and steri li zed spent hop s afte r 4 days incubation in th e dark and 3 6 hour s in the l ig ht ( Hu szcza, 2008). Hops (Humulu s s pp .) ha ve been ob served t o g rown as a wetland spec ies in North Caro lina , indi cat ing th at the re may be so me level of natural backgro und exposu r e of aqu at ic o r ganis m s to h ops be ta acids (Krin gs, 2 002). Conclusion: Base d up o n a lack of ex p os ure to aqu at i c e nvironme nt s v ia th e u se of t he end-use produc t in beehi ves, no adve rse e ffect s are ex pec te d in aquatic o rganisms as a r es ult of the u se of HopGuard ® (16

. 0% potass ium sa lts ofhop s be ta ac id s). There are no concerns for non -target aquatic organisms. C lassification: ACCEPTABLE

Terrestrial Plant Toxicity, Seedling Emergence & Vegetative Vigor (O CS PP 850.4100 & 850.4150): It is hi gh ly unlikel y that t h ere will be any ex po s ure of pota ss ium s alt s of hop s b e ta ac id s to plants v ia the app li cat i o n ofHopG u ard ® ( 16.0 % a .i. ) to the in s i des o f commerc ial bee hi ves . In the unlike ly event that hop s beta ac ids woul d migrate outside the beehive , th ey wo uld be rapidl y degraded by so il fungi. In labo ratory experiments , Tra m e tes versicolo r was 2 1Hop Beta Acids PC Codes: 005115 Review Typ e : Re s idue Studie s, Tier I Tox , Tier I Non-targets DP Numbers : 417758 & 417759 E PA Fil e No. : 83623-G , -R, & -E

shown to fully degrade humulones and Ju pu lones in commercial hop extract and sterilized spent hops after 4 days incubation in the dark and 36 hours in the light (Huszcza , 2008). Based upon a lack of exposure to terrestrial environments via the use of the end-use product in beehives , no adverse effects are expected in plants as a result ofthe use ofHopGuard ® (16.0% potassium salt s of hops beta acids). There are no concerns for non-target plants. Non- Target Insect Testing (OCSPP 880.4350): 

### It is highly unlikely that there will be any exposure of potassium salts of Hops Beta Acids to non-target insects via the application of HopGuard® ( 16.0% a.i.) to the insides of commercial beehives

. Three years of residue studies using Hops Beta Acids in beehives to control Varroa mites demonstrated that there were no adverse effects on bees. It is highly unlikely that non-target insects will gain access to the treated strips inside the hive due to the vigorous defense of hives by the honey bees again st invaders (Breed et. al., 2004).
QUALITATIVE ENVIRONMENTAL RISK ASSESSMENT Non -target organism studies and environmental fate data were not submitted for review. In lieu of Gu idel in e studies , the registrant submitted rationales in support of the Tier I non-target organism data requirements. Based on a lack of exposure , effects of Hops Beta Acids on non-target organisms , including threatened and endangered spec ies , are highly unlikely and not anticipated

. Runoff into ground and surface waters is not anticipated. The product (plastic strips treated with a 16% solution of Hops Beta Acids) is manually inserted into beehives. It is highly unlikely that organisms external to the hive wi ll have access to the treated strips , and little or no likelihood that the active ingredient will migrate from the hive to the external environment. In the highly unlikely event that Hops Beta Acids would migrate out of the beehive to the external environment , environmental fate data indicate that Hops Beta Acids wou ld degrade rapidly, with 100% degradation in 36 hours in the light and 4 days in the dark (Huszcza , 2008)

.

22Hop Beta Ac id s PC Codes : 005115 Review Type: Residue Studies , Tier I Tox , Tier I Non-targets DP Numbers: 417758 & 4 17759 EPA File No.: 83623-G, -R, & -E


### CONCLUS IONS: There are no co nc e rn s for non-target organisms, in cluding threatened and e ndan gered spec ies w hen the product is used in accordance with EPA-approve d product labe ling. B ased on littl e o r no li ke lihood of exposure to any non- target o r gan is m , the Agency makes a "No Effect" determination for threatened and endangered spec ies. 

### NO DERs WERE WRITTEN FOR THIS REVIEW

cc: R. S. Jones , M. Adams , BPPD Subject F ile/ BPB Fi l e/ IHAD/ Archives R. S. Jones, Ph.D. , Sr. Biologist , FT , OPY: 12 / 04 /20 14



### 23Hop B e ta Acids PC Codes: 0051 15 Review Type: R esidue Studies , T ier I Tox , Tier I Non-target s REFERE NCES DP Numbers: 41775 8 & 417759 EPA Fi l e No.: 83623

- G, -R, & -E

Breed , M.D. , E. Guz man- Nova, and G. J

. hun t. 2004. Defensive be hav ior of honey bee s: organ ization , ge n eti cs, and comparisons with other bee s . Annual Review of E ntomo l ogy 49: 27 1 -296. C hin , Y -C. , and H

. H. A nd e rson. 1 950. To x ico logy and pharmacology of lupulon. Arch

. Int. Pharmaco dy n. 82(1 ): 1-15. C lee mput , M. V. , A. H eye ri ck, C. Lib ert, K. Swerts, J. Philipp e , D

. D. Keukeleire, G

. Haege man , a nd K. D. Bo ssc he r. 20 09. Hop bitter ac id s ef fici e ntl y bl oc k inflanunation ind epe nd ent of ORa lph a, PPAR-alpha , or PPAR-gamma. Mol ec ular Nutrition and Food R esea rch 53: 114 3 -11 55.
D o ng, N. T. K and R. B. Ogle

. 2003. Effect of brewery waste r ep l aceme nt of concentrate on the p e rformance of l oca l and crossbred growin g Muscovy duck s. Asian-Australian Journal of Animal Scie nce 16(10): 1510

- 1517. Dum as , E

. R ., A. E. Mc haud , C. B. B e r ge ro n , J. L. LaFrance, S

. Mortillo , and S. Gafn er. 2009. Deodorant effects of a supercritical hops extract: antibacterial act ivi t y against 

### Coryneb act erium xerosis and 

### S ta phylococcus e pidermidis and efficacy te s ti ng of a hop s/z inc ri c in oleate stick in humans th rough the se n sory eva luation of axillary deodorancy. Journal of Cosmet ic Dermatology 8: 197-204. Estrada , J. L., F. Gorzalo , C

. Cec chini , and E. Casqueta. 2002. Co nt ac t urti caria from hop s 

### (Humulus lu pus) in a p atie nt with previou s urticaria-an g io de ma fro m peanut , c hestnut, and b a nana. Co ntac t D e rm at iti s 48 : 12 7. FDA , 200 1

. Age ncy re spon se letter GRAS Notice No. G RN 000063. C FS AN/Office of Pre m a rk et App r oval. March 15 , 2001. Hu szcza , E. , A. Bartman ska , M

. Aniol , W. Maczka, A. Zo lni e rczyk , and C

. Wawrzenczyk. 2008. Deg rad ation of hop bitter acid by fun gi. Wa s te m an ageme nt 28(8): 140 6-1 410. Krin gs, A. 2002. Keys to t h e vines of Caro li na wetland s. Vulp ia 1: 23

- 40.
Lamy, V. , S. Rous s l , M. Chaabi, F. Gosse ', N

. Schall , A. Lobstein, and F. R aul. 2007

. Carc in oge nes i s 28 : 1575-1581. Moir , M. M. 20 00. J ou rn a l of th e American Soc iety of Brewery C he mi s t s 58(4) : 1 3 1-146.

24Hop B eta Acid s P

C Codes: 0051 15 Review Type: R es idue Studies , Tier I Tox , Tier I Non-targets DP Numbers: 417758 & 417759 EPA File No.: 83623-G , 

### -R. & -E

National Hon ey B oard. 2014. Honey Industry Facts. Accessed 12/ 02/2014. http: //www .honey .com/news room/press-kit s/honey-indu stry- fact s Nat io n a l Ho t dog and Sausage Council (N HDSC)

. 20 14

. Hot Dog Fast Facts. Accessed 12/ 04/ 20 14. http: //www.hot-dog.org/ culture/hot-dog-fa s t-f ac t s Nutrition Fact s . 20 14. Frankfurter , beef[frank , hot dog , weiner]. Accessed 12/ 04/ 2014. http: // nutri tiondata.self.com/facts/ sausages-and-luncheon-meats/ 1 3 3 8 / 2 S iragusa , G. R. , G. 1. Haas, P. D. Matthews , R. J. Smith, R. J. Buhr, N. M. Dale , and M.G. Wise. 2008. Antimicrobia l activity of lupu lone against C lo s tridium perf rin gens in the ch icken inte s tinal tractjejunwn and caecwn . Journal of Antimicrobial C hem o th erap y 61: 853-858. S hen , C

. , I. Gcornaras , P. A. Kendall , and 1. N. Sofos. 2009. Co ntrol of 

### Listeria monocytogenes on Frankfurters b y dipping in hops beta acids so luti o n s. Journ al of Food Protection 72(4): 702- 706. U SD AIF S IS. 20 1 3. FSIS Directi ve 7120.1 , Revision 15. Sa fe and Suitable Ingredients Used in the Production of Meat. Pou ltry and Egg Products. Dated 04 113 / 20 1 3. Verze le , M. and D. De Keuke1ele. 1991. Chemistry and Analys i s of Hop and Beer Bitter Acids. E lsevier. Am s t erdam.

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