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

MEMORANDUM

DATE: June 12,2014 OFFICE OF CHEMICAL SAFETY

AND POLLUTION PREVENTION

SUBJECT: Macleaya Extract: Preliminary Risk Assessment for Registration Review

PC Code: 069095

Decision No.: 489 178

P etition No.: Not Applicable (N/ 

### A)

Risk Assessment Type: Qualitative

TXRNo.:N/A

MRID No.: N/A DP

. Barcode: 4 18 970 Registration No.: 69876-1 Regulatory Action: Registration Review

Case No.: 7024

CAS No.: 112025-60-2

40 CFR: N/A

FROM: Jessica Kidwell, Ris k Assessor 

### WVU.<!.W

# ~ ........ 

## A.
Kelly O'Rourke,

## Occupational/Resi~ntial**Ex sure** **A~sessor "f** **'...L~** 

# v{/, Q

.. Abdallah Khasawinah, Ph.D

. , Tox i cologist /

### {/~ _)

RAB IV

, Health Effects Division (7509P)

THROUGH: Elissa Reaves, Ph.D., Branch Chief

# ?"~~**2c:: ---..** **-­** RAB IV, He alt h Effects Division

## (7509~

TO: Susan Bartow, Chemical Review Ma n ager Kevin Costello

, Act ing Branch Chief

RMIB IV, Pesticide Re-Evaluation Division (7508P)

Attac h ed i s the Health Effect Division's (HED's) human health r i sk assessme n t fo r macleaya

extract to support registration review. A screening level occupational assessment did not identify any potential risks of concern via dermal or inhalation routes of exposure; the margins

of exposure (MOEs) for dermal and inhalation route$ did not fall below the Agency's leve l of

concern (LOC). A dietary risk assessment is not required because macleaya extract has no registered food uses

. A drinking water exposure assessment is not r equired based on l abel specifications that li mit use to enclosed commercial greenhouses. In addition, the

Environmental Fate and Effects Division (EFED) an t icipates that no quantitative drinking

water exposure assessment will be n eeded. Based on th e r:egistered uses, th ere are no r es iden tial

exposures for mac l eaya extract, therefore, a resident ial exposure assessment is no t applicable.

### Macleaya Extract/069095 Human Health Risk Assessment D418970

Introduction

Macleaya extract is t he act i ve i ngredient (1.5%) in the product Qwel TM 

### (CTI 13 -1 9B). It 

### is an ornamental plant fungicide used in enclosed comme r cial greenhouses to contro l powdery

mi ld ews and l eaf spots. Macleaya extract i s formulated as a liquid concentrate and applied as a foliar spray by backpack or h i gh pressure sprayer. The maximum application rate is 5 fl

. oz. formulation/! 0 gallons water applied t o 1 ,25

. 0 ft 2 

### for roses and other larger plants (i.e.

, 0.17 pounds active ingredient per acre (lb ai / A) or 0.0005 lb a. i

. per gallon). Foliar applications may be made at 8- 1 0 day intervals as needed. The label does not specify t he number of applications. Macleaya extract has no registered residential uses; residential exposures are, therefore, not expected. Usage of macleaya extract was not reported in the Nationa l Agr i cul t ural Statistics

Serv i ce (NASS) nursery and floriculture usage surveys conducted in 2002, 2004, or 2007 nor

was it reported in the 2006

- 2008 California D epartment of Pes t icide Regulation (CALDPR) pestic i de usage reports (BEAD Memo, Stebbins

, K, Yourman, L. 

### and Fau l kner, 12/7 / 2010).

Hazard Identification and Dose Response Assessment

Macleaya extrac t is a fungicide used for the contro l of fo li ar fungal diseases, such as powdery mildew, alternar i a leaf spot and septaria l eaf spot on ornamenta l plan t s grown in greenhouses. It



### i s the extract of perennia l p l ants Macleaya cordata and Macleaya microcarpa (Macleaya spp.).

R aw macleaya extract is a mixture of alkaloids be l onging to t he quaternary benzophenathridine

alkaloid (QBA) Class. The purified product conta i ns two princ i pal QBAs, sanguinarine (47

- 53%) and chelerythrine (20

- 26% )

.

The t oxicology database for macleaya extract is very lim i ted and i s supported by some stud i es on the closely related sanguinaria extract (contains approximately 3 7

- 40% sanguinarine, 16

- 18%

che l erythrine, and other related products rang i ng from less than 1% to 9% of the total composition), and sanguinari n e chloride, a major component of macleaya extract. Based on a comparison of s t ruc t ural similarities, physical and chemical characteristics and toxicological

information found i n l i terature studies, HE D determ i ned that available tox i city stud i es on sanguinaria extract and sanguinarine chloride are l ikely to be acceptable for use as surrogate data

fo r macleaya extract. The tox i cology database for macleaya extract (based on the use of
surrogate data) is adequate for registration review. T he availab l e stud i es a r e listed below and i n Appendix A.

• Acute: Oral, dermal, inhalation on macleaya extract. Acute oral and derma l on sanguinaria extract. Acute oral, inhalation, eye irritation, dermal i rritation, and skin

se n s i t i zation on sanguina r ine chlor i de.

• Subchronic: Sangu i naria extract/sangui n a r ia chloride, 90-day oral tox i c i ty (rat, monkey)

.

• Developmental t oxicity: Sanguinaria extract, developmental tox i c i ty (rat and rabb i t)

• Reproduction: Sanguina ri a extract, !-generat i on reproduction (rat)

• Chronic : Sanguinaria extract, combined oral chronic toxicity/carcinogenicity (rat).

• Other : Sangu i naria extract/sanguinaria chloride, mutagen i city battery

.



### Al t hough the 201 1 scop i ng document (D384098) had previously recommended multiple studies

be submitted (90-Day Inha l ation Toxicity Study, 90-Day Dermal T oxicity Study

, Developmental

Page 2 

### o f 17

### Macleaya Extract/069095 Human Health Risk Assessment D418970

Page 3 of 17 Toxicity Study in Rabbit s , Developmental Toxicity Study in Rat s
, 2

- Generation Reproduction Toxicity in Rat s , Acute Neurotoxicity Study in Rat s , Subchronic Neurotoxicity Study in Rat s)

, HED’s Hazard and Science Policy Council (HASPOC) w aived the requirements for these studies

in 2013 (TXR 0056588 )

. The HASPOC used the Weight of Evidence ( WOE ) in waiving the requirements for the se studies based on the following : 1) the limited exposure prof ile of macleaya extract; 2) there are no registered food uses of macleaya extract at this time ; 3) no exposure to macleaya extract is expected through drinking water ; 4) the use of an oral point of departure ( POD ) results in dermal MOEs that d o not fall below HED’s level of concern ( target
LOC is for MOEs < 300) for highest occupational exposures ( i.e., MOEs are greater than 3,600 and 440 for handlers and post

- application workers, respectively ); 5 ) the use of an oral POD

results in inhalation MO Es that do not fall below HED’s level of concern for workers (MOE greater than 13,000 ); and 6 ) the low vapor pressure (3.92 x10

- 11 

### mm Hg at 25  C) and physical chemical properties of s anguinarine , a major component of macleaya extract. HASPOC also

waived th e immunotoxicity study data requirement (TXR No. 0056719 )

.

Sanginaria is available commercially in various over

- the

- counter products. Sanginaria extract is

used in Viadent ® 

### oral rinse and toothpaste products (Frankos et al

. 1990). Sanguinaria extract

also is known to treat gum disease (removes plaque), athlete’s foot, chronic cough, asthma, ringworm, rashes, and fungus on the skin when applied topically.

Macleaya extract exhibits low acute toxicity via oral and dermal rou tes of exposure. It is not a dermal irritant. Primary eye irritation and dermal sensitization studies are available on the

formulation but not on the technical material.

Available chronic, subchronic, developmental, and reproductive studies on surrogate sanguinaria compounds are available primarily from the published literature. Original reports of the published studies have been submitted, but not yet reviewed. A preliminary screen of the study reports indicates concordance with the conclusions of the p ublished studies. The test material in all these studies (developmental, subchronic, chronic, reproduction) was administered by gavage. These studies indicate that macleaya extract is likely to be irrita ting to the stomach and lungs. Subchronic and chronic oral studies in monkeys and rats

, respectively

, showed gastrointestinal tract effects indicative of irritation. The subchronic study in rats reported lung

abnormalities indicative of inflammation. Clinical signs of rales, breathing difficulty, excessive

s alivation, lethargy, reduced motor activity were observed in parental animals in developmental and reproductive toxi city studies in rats. No developmental effects were observed in a developmental rat study at maternally toxic doses. In the developmental ra bbit study, fetal effects (e.g., decreased number of fetus/litter) were observed at maternally toxic doses. No reproductive effects were observed in the rat reproduction study.

Sanguinarine c hloride showed equivocal positive response in mammalian cell f orward mutation assay in the presence of microsomal activation system, was negative in in vitro mammalian cytogenic test system, equivocal response in a DNA repair system. Sanguinaria extract was negative in the E. coli mutation test, but produced positive response in Salmonella strains TA98, TA100, TA1537, TA1538 (+S9). Sanguinaria e xtract was not mutagenic in Salmonella strains TA98, TA100, TA1535, TA1537, TA1538 (

- S9) and TA1535 (+S9). Sanguinaria e xtract was negative in a chromosomal aberration assay and a DNA repair system. Macleaya extract,

### Macleaya Extract/069095 Human Health Risk Assessment D418970

Page 4 of 17 sanguinaria extract, and sanguinarine chloride have not been evaluated for carcinogenicity. A published article demonstrated that Macleaya Cordata extract administered as a dietary

admixture to rats at 600 mg/kg f eed induced no DNA damage to rat lymphocytes or hepatocytes

after 90

- days of oral administration (Stiborova, M. et al., 2008)

At the time the scoping document was prepared in 2011, t here were no macleay a extract cases in the main or aggregate Incident Data Syste m (IDS) databases; therefore, there did not appear to be

a concern at th at time that would warrant further investigation (Recore, 11/30/10, D384109)

. A n updated IDS query for macleaya extract also indicated there were no incidents reported to either

Main or Aggregate IDS (Email correspondence, S. Recore, 4/29/14).

There are no registered food, feed or residential uses for macleaya extract. T herefore

, toxicity PODs were not selected for dietary or r esidential exposures. PODs were selected for occupational exposures via dermal and inhalation pathways for use in the 2002 occupational exposure and risk assessment based on registered uses. The 2002 occupational exposure

assessment relied on the use of en dpoints of toxicity derived from the one

- generation reproduction study in rats and the subchronic oral toxicit y study in monkeys. D uring its deliberations on waiving various studies , the HASPOC recommended that the endpoint of toxicity derived from the sub chronic oral toxicity study in monkey be used for assessing both the dermal and inhalation exposures ( short

- and intermediate

- term durations) . In this study, monkeys were orally gavaged with sanguinarine chloride at 0, 10 , 30 or 60 mg/kg/day. The LOAEL was 10 mg/kg/day (lowest dose tested) based on diarrhea and emesis

. T he default 10 X factor f or extrapolation from a LOAEL to NOAEL was reduced from 10X to 3X since the test animal was a monkey. The total uncertainty factor is 300X (10X for intraspecies variat ion, 10 X for interspecies extrapolation, and 3X for use of a LOAEL). Since no inhalation absorption data are available, toxicity by the inhalation route is considered to be equivalent to the estimated toxicity by the oral route of exposure (100% absorption factor).

T here are no dermal absorption studies or ADME available for this product. Raw macleaya extract is a mixture of alkaloids belonging to the quaternary benzophenathridine alkaloid (QBA) Class. The purified product contains two principal QBAs, san guinarine (47

- 53%) and chelerythrine (20

- 26%). These are charged molecules and are not expected to be absorbed through the skin. Experimental data in humans have demonstrated that the quaternary chemical diquat (with chemical structure resembling the macle aya extract components) was minimally absorbed with only 0.3% absorbed and excreted over a 5 day period (Feldman and Maibach,

1974

, MRID 43736616) . This human study was subjected to the appropriate ethical evaluation by the Agency (Memo from John Carley to Anne Overstreet, July 27, 2006). Thus for macleaya extract, the dermal absorption is expected to be low and to allow for lack of a dermal absorption study, therefore, a 10% dermal absorption factor is proposed for this risk assessment and is

considered to be a conservative assumption

. See Appendix B for Endpoint Selection.

Sanguinarine C helerythrine Diquat

### Macleaya Extract/069095 Human Health Risk Assessment D418970

Page 5 of 17 Dietary Exposures

There are no food uses for macleaya ext r act , so exposure from consumption of food is not expected . Therefore, there is no dietary risk

.

Occupational and Residential Exposures

Based on the registered uses, there are no residential exposures for macleay a extract, therefore,

there is no residential risk.

Although exposure may occur during occupational handling and post

- application activities, a screening level assessment based on maximum application rates and baseline attire (additional

PPE [i.e., gloves ] are required on the label) indicated no risk of con cern via dermal and

inhalation routes of exposure. Estimates of occupational handler and post

- application

exposure /risk are summarized in Tables 1 and 2, respectively. Furthermore, t he current label requires all handlers to wear personal protective equipme nt (i.e., chemical

- resistant gloves in addition to baseline attire) and specifies a restricted entry interval of 12 hours

, which is considered to be adequately protective.

### Macleaya Extract/069095 Human Health Risk Assessment D418970

Page 6 of 17 Table 1

. Summary of Short

- /Intermediate

- Term Handler Exposures/Risks for Macleaya Extract

Exposure
Scenario Max

Applica

tion Rate 1 Unit Exposure 2

(mg/lb ai) A rea

Treated

or Amount

Handled daily 3 Absorbed Dose

(mg/kg/day) 4 MOE 5

Total
MOE 6 Dermal Inhal a
tion Dermal Inhal a
tion Dermal Inhal ation

GREENHOUSE ORNAMENTALS

- MIXER/ LOADER (single layer, no gloves )

Groundboom 0.17

lb ai/A 0.22 0.00022 60 A 0.00048 0.000028 3,600 360,000 3,600

GREENHOUSE ORNAMENTALS

- APPLICATOR (single layer, no gloves)

Groundboom 0.17

lb ai/A 0.0786 0.00034 60 A 0.00021 0.000043 10,000 230,000 9,600

GREENHOUSE ORNAMENTALS

- MIXER/ LOADER /APPLICATOR (single layer, no gloves) Backpack

0.0005

lb ai/gal 13.2 0.14 40 gal 0.00028 0.000035 30,000 290,000 27,000 Manually

pressurized

Handwand 100 0.030 40 gal 0.000011 0.0000075 4,000 1,300,000 4,000

Mechanically

pressurized Handgun 3.5 0.12 1,000 gal 0.0016 0.00075 4,600 13,000 3,400

1 Application r ate based on label for macleaya extract (EPA Reg. No. 69876

- 1).

2 Based on HED Exposure Science Advisory Council ’s Occupational Pesticide Handler Unit Exposure Surrogate Reference Table

. 3 HED Exposure Science Advisory Council Policy #9.1

4 Daily Absorbed Dose (mg/kg/day) = daily unit exposure (mg/ lb ai) × application rate (lb ai/ A or gal ) × a rea treated ( A or gal/ day) × a bsorption ( 10 % for dermal) ÷ body weight (80 kg )

. 5 MOE = L OAEL (dermal = 10 mg/kg/day for short

- /intermediate

- term exposure; inhalation = 10 mg/kg/day for short

- /intermediate

- term ) ÷

a bsorbed d ose (mg/kg/day). LOC = 300.

6 Total MOE = 1 ÷ [(1 ÷ Dermal MOE) + (1 ÷ Inhalation MOE)]

Table 2. Post

- application Exposure and Risk for Macleaya Extract

Crop Applicatio

n Rate (lb ai/acre) Activity Exposure
Time (hrs/day) Transfer

Coefficient 1 (cm 2 /hr) Day 0

DFR 2 (ug/cm 2 ) Daily

Dermal

Dose 3 (mg/kg

- day) MOE 4

Greenhouse Crop (Ornamentals,

Non

- bearing Plants) 0.17 Harvesting, Hand; Pruning, Hand; Scouting; Container Moving; Weeding, Hand; Transplanting; Grafting; Propagating; Pinching; Tying/Training 8 230

0.48 0.0011 9,100

Cut Flowers Hand Harvesting 4,800 0.023 440

1 Transfer coefficient from HED Exposure Sc ience Advisory Council Policy #3

2 D islodgeable F oliar R esidue (DFR) on the day of application (i.e., “day 0”) = a pplication r ate × percent of residue available for transfer

(25%) × 4.54E8 μg/lb × 2.47E

- 8 acre/cm 2

3 Daily Dermal Dose = [DFR (μg/cm 2 ) × t ransfer c oefficient × 0.001 mg/μg × 8 hrs/day × dermal absorption ( 10 %)]  BW ( 80 kg).

4 MOE = LOAEL ( 10 mg/kg/day) / d aily d ermal d ose. LOC = 300.

### Macleaya Extract/069095 Human Health Risk Assessment D418970

Page 7 of 17 Volatilization Based on the Agency's current practices, a quantitative post

- application inhalation exposure assessment was not performed for macleaya extract at this time primarily because of the very low vapor pressure ( 3.92 x10

- 11 

### mm Hg at 25  C ) ) . Although a quantitative post

- application inhalation exposure assessment was not performed, a screening

- level inhalation exposure assessment was performed for handlers. This exposure scenario is representative of a worse case inhalation exposure and sho uld be considered protective of most post

- application inhalation exposure scenarios.

Spray Drift The registered use for macleaya extract is limited to ornamentals in greenhouses , therefore, this
use pattern will not result in spray drift outdoors

.

Endo crine Disruptor Screening Program

As required by FIFRA and FFDCA, EPA reviews numerous studies to assess potential adverse

outcom es from exposure to chemicals. Collectively, these studies include acute, subchronic and chronic toxicity, including assessmen ts of carcinogenicity, neurotoxicity, developmental, reproductive, and general or systemic toxicity. These studies include endpoints which may be susceptible to endocrine influence, including effects on endocrine target organ histopathology, organ weights, estrus cyclicity, sexual maturation, fertility, pregnancy rates, reproductive loss, and sex ratios in offspring. For ecological hazard assessments, EPA evaluates acute tests and chronic studies that assess growth, developmental and reproductive effects in different taxonomic groups. As part of its preliminary risk assessment for registration review of macleay a

extract

, EPA reviewed the available data and selected the most sensitive endpoints for relevant risk assessment scenarios from the existing hazard database. However, as required by FFDCA section 408(p), macleay a extract is subject to the endocrine screening part of the Endocrine Disruptor Screening Program (EDSP).

EPA has developed the EDSP to determine whether certain substances (including pesticid e active and other ingredients) may have an effect in humans or wildlife similar to an effect produced by a “naturally occurring estrogen, or other such endocrine effects as the Administrator may designate.” The EDSP employs a two

- tiered approach to making the statutorily required determinations. Tier 1 consists of a battery of 11 screening assays to identify the potential of a chemical substance to interact with the estrogen, androgen, or thyroid (E, A, or T) hormonal systems. Chemicals that go through Ti er 1 screening and are found to have the potential to interact with E, A, or T hormonal systems will proceed to the next stage of the EDSP where EPA will determine which, if any, of the Tier 2 tests are necessary based on the available data. T ier 2 testing is designed to identify any adverse endocrine

- related effects caused by the substance, and

establish a dose

- response relationship between the d ose and the E, A, or T effect. Under FFDCA section 408(p), the Agency must screen all pesticide chemicals. Betwe en October 2009 and February 2010, EPA issued test orders/data call

- ins for the first group of 67 chemicals, which contains 58 pesticide active ingredients and 9 inert ingredients. A second list of chemicals identified for EDSP screening was published on J une 14, 2013 ( http://www.regulations.gov/#!documentDetail;D=EPA

- HQ

- OPPT

- 2009

- 0477

- 0074 ) and includes some pesticides scheduled for registration review and chemicals found in water

. Neither

### Macleaya Extract/069095 Human Health Risk Assessment D418970

Page 8 of 17 of these lists should be construed as a list of known or likely end ocrine disruptors. For further information on the status of the EDSP, the policies and procedures, the lists of chemicals, future lists, the test guidelines and the Tier 1 screening battery, please visit our website ( http://www.epa.gov/endo/ )

.

Data Requirements

None. The stu dies required as part of the 2011 HED scoping document were waived by

HASPOC in 2013 (TXR Nos. 0056 588 and 0056 719 )

.

Conclusions

A screening

- level exposure assessment did not identify potential risks via dermal or inhalation

routes resulting from occupational exposure scenarios . The conservative dermal and inhalation

MOEs are not of concern. A d ietary risk assessment is not required because mac leaya extract has no registered food uses and drinking water exposure is not anticipated based on label specifications that limit use to commercial greenhouses

. There is also no residential exposure

based on the label uses. Therefore, based on the lack of exposure, there are no dietary or

residential risks for macleaya extract

.

References

Collantes, M. 8/9/2002. D284631. Occupational and Residential Exposure and Risk for the Proposed Use of Macleaya Extract in Enclosed Comm ercial Greenhouses.

Daiss, B. 1/5/2011. D384098. Macleaya Extract: Human Health Risk Scoping Document in Support of Registration Review.

Feldman and Maibach, 1974. Percutaneous penetration of some pesticides and herbicides in man. Toxicology and Applied Pharmacology 28: 126

- 132; MRID 43736616.

Frankos , V. et al. 1990. Safety of sanguinaria extract as used in commercial toothpaste and oral rinse products. J. Can. Dent. Assn. 56:41

- 47.

Habiba, U. 7/18/2013. TXR No. 0056719. Macleaya Extract: Summary of H azard and Science Policy Council (HASPOC) Meeting of July 18, 2013: Recommendation on the Need for Immunotoxicity Study (870.7800).

Hurley, P. 7/30/2002. TXR No. 0050979. Macleaya Extract

– 1 st 

### Report of the Hazard

Identification Assessment Review Committ ee.

Recore, S. 11/30/2010. D384109. Macleaya Extract: Review of Human Incidents.

Recore, S. 4/29/2014. Email correspondence from S. Recore to J. Kidwell.

### Macleaya Extract/069095 Human Health Risk Assessment D418970

Page 9 of 17 Rury, K. 3/5/2013. TXR No . 0056588. Macleaya Extract: Summary of Hazard and Science Policy Council (HASPOC) Meeting of February 14, 2013: Recommendations on the Need for Multiple Studies.

Stebbins, K. Yourman, L. Faulkner, J. 12/7/ 2010. BEAD Chemical Profile for Registration Review: Macleaya Extract.

Stiborova, M. et al. 2008. Macleaya Cordata Extract and Sangrovit Genotoxicity Assessment In

Vivo . Biomed. Pap. Med. Fac. Univ. Palacky Olomouc Czech Repub. 152:35

- 39.

### Macleaya Extract/069095 Human Health Risk Assessment D418970

Page 10 of 17 APPENDI X A

A. TOXICOLOGY DATA SUMMARY

A.1. Macleaya Extract Guideline Toxicology Data Requirements: The toxicology data requirements (40 CFR 158. 500 ) for non

- food use are presented in Table A.1.1. Use of the new guideline numbers does not imply that the new (1998) guideline protocols were used.

Table A.1.1. Macleaya Extract Guideline Data Requirements

Study Technical

Required Satisfied

870.1100 Acute Oral Toxicity

................................ ................................ .. 870.1200 Acute Dermal Toxicity

................................ ............................. 870.1300 Acute Inhalation Toxicity

................................ ......................... 870.2400 Primary Eye Irritation

................................ ............................... 870.2500 Primary Dermal Irritation

................................ ......................... 870.2600 Dermal Sensitization

................................ ................................ . yes

yes

yes

yes

yes

yes yes

yes

yes

yes

yes

yes

870.3100 Oral Subchronic (rodent)

................................ .......................... 870.3150 Oral Subchronic (nonrodent)

................................ .................... 870.3200 21/28

- Day Dermal

................................ ................................ .... 870.3465 21/90

- Day Inhalation

................................ ................................ y es (CR)

yes (CR) yes* (CR) yes* (CR) -

-

-

-
870.3700a Developmental Toxicity (rodent)

................................ .............. 870.3700b Developmental Toxicity (nonrodent)

................................ ........ 870.3800 Reproduction

................................ ................................ ............ Yes*

Yes*

Yes* -

-

-
870.4100a Chronic Toxicity (rodent)

................................ ......................... 870.4200a Carcinogenicity (rat)

................................ ................................ . 870.4200b Carcinogenicity (mouse)

................................ ........................... 870.4300 Chronic/ Carcinogenicity

................................ .......................... yes (CR)

yes (CR)

yes (CR)

yes (CR) yes

yes

yes

yes

870.5100 Mutagenicity

— Gene Mutation

- bacterial

................................ 870.5300 Mutagenicity

— Gene Mutation

- mammalian

........................... 870.5375 Mutagenicity

— Structural Chromosomal Aberrations

.............. 870.5900 Mutagenicity

— Other Genotoxic Effects

................................ .. yes

yes

yes

yes yes

yes

yes

yes

870.6100a Acute Delayed Neurotox. (hen)

................................ ................ 870.6100b 90

- Day Neurotoxicity (hen)

................................ ...................... 870.6200a Acute Neurotox. Screening Battery (rat)

................................ ... 870.6200b 90

- Day Neuro. Screening Battery (rat)

................................ ..... 870.6300 Develop. Neuro

................................ ................................ ......... no

no yes* yes*

no -

-

-

- yes

870.7485 General Metabolism

................................ ................................ . 870.7600 Dermal Penetration

................................ ................................ ... 870.7800 Immunotoxicity

................................ ................................ ........ no

no yes* * yes

yes

-

* Waived (HASPOC report TXR # 0056588)
**Waived (HASPOX report TXR #0056719) CR: CONDITIONALLY REQUIRED

### Macleaya Extract/069095 Human Health Risk Assessment D418970

Page 11 of 17 A.2 Toxicity Profiles

Table A.2. 1: Acute Toxicity Profile of Macleaya Extract

GDLN Study Type MRID Results Tox Category

81

- 1 Acute Oral 44525106 Males:= 1016 mg/kg in corn oil); 1544 mg/kg (CMC) Females:= 629 mg/kg in corn oil; 960 mg/kg (CMC) III

81

- 2 Acute Dermal

- rabbit 44525107 (1997) LD 50 > 2000 mg/kg III

81

- 3 Acute Inhalation 44525108 (1997) Males : < 0.22 mg/L

Females:> 0.22 mg/L and < 0.52 mg/L II

81

- 4 Primary Eye Irritation

of macleaya end use product formulation 44525109 (1998) Eye irritant I

81

- 5 Primary Skin Irritation

end use product
formulation 44525110

(1997) Not irritating IV

81

- 6 Dermal Sensitization

- Not conducted N/A

Table A.2.2 : Acute Toxicity Profile of Sanguinaria Extract

GDLN Study Type MRID Results Tox Category

81

- 1 Acute Oral 44525111

Literature

summary 1440 mg/kg in water 1250 mg/kg in corn oil III

81

- 2 Acute Dermal

- rabbit 44525111

Literature

summary

46273804 (1983) LD 50 > 200 mg/kg II

81

- 3 Acute Inhalation

- Not conducted

-

81

- 4 Primary Eye Irritation

- Not conducted

-

81

- 5 Primary Skin Irritation

- Not conducted

-

81

- 6 Dermal Sensitization

- Not conducted N/A

### Macleaya Extract/069095 Human Health Risk Assessment D418970

Page 12 of 17 Table A.2.3 : Acute Toxicity Profile of Sanguinarine Chloride

GDLN Study Type MRID Results Tox Category

81

- 1 Acute Oral 44525111

Literature

summary LD 50 = 1525 mg/kg in CMC 1664 mg/kg in corn oil III

81

- 2 Acute Dermal

- rabbit

- Not conducted.

-

81

- 3 Acute Inhalation 44525111

Literature

summary Mild pulmonary and ocular irritant

0.16

- 0.37 mg/mL, LC 50 unclear. Conservative

estimate of 0.16 mg/mL II?

Acute Inhalation 46273806 (1983) Males: < 0.22 mg/L

Females :> 0.22 mg/L II

81

- 4 Primary Eye Irritation 44525111

Literature

summary Mild irritant cannot
estimate

81

- 5 Primary Skin Irritation 44525111

Literature

summary Slight irritant cannot
estimate

81

- 6 Dermal Sensitization 46273803 (1990) Not a sensitizer

-

### Macleaya Extract/069095 Human Health Risk Assessment D418970

Page 13 of 17 Table A. 2.4 Subchronic, Chronic and Other Toxicity Profile for Macleaya Extract

Guideline No. Study Type MRID No. (year)

Classification

Dose Levels Results

28

- day dietary toxicity study

- rat sanguinaria extract (33.1%, sanguinarine chloride and 68.4%

total alkaloid ) 45400501
Unacceptable/non

guideline

0, 5, 15, 45, 135, and 405 mg/kg/day NOAEL = 405 mg/kg/day (HDT)

LOAEL was not observed

870.3100

13

- w eek feeding

- rat ( s anguinaria

e xtract: (44.5% sanguinarine chloride and 78.6% alkaloid)) 45400502 (1987)
Unacceptable guideline.

0, 50, 100, 200, 300, or

400 mg/kg/day by gavage (20 mL/kg) NOAEL : not established LOAEL: 50 mg/kg/day (decreased overall body weight gain, labored breathing and rales in both sexes (LDT)).

Blood clotting measurements were not quan tified.
The nose, pharynx, larynx, and seminal vesicles were not examined histologically.
There was a very high rate of death and gavage error incidences

which tended to confound the results. Insufficient numbers of animals were available in some groups to allow for an adequate interpretation of the results.

4

- week range finding study
cynomolgus monkey
( s anguinaria extract (purity not provided) 45400503 (1987)
Acceptable/non

guideline

O, 50, 100, 150, 200 mg/kg/day in O.2% CMC 2 mL/kg
One monkey/sex/dose NOAEL : 50

LOAEL: 100 mg/kg/day increased and/or severity of clinical signs (emesis, diarrhea, inappetence, and dehydration).

870.3100

13

- w eek cynomolgus monkey ( s anguinarine c hloride (98.8% a.i) 45400504 (1988)
Acceptable guideline

0, 10, 30, or 60 mg/kg bw/day by gavage i n 0.1% CMC (2.0 mL/kg)

4 monk e y s/sex/dose NOAEL : not established

LOAEL: 10 mg/kg/day (increased incidence (number of affected animals) and frequency of emesis and diarrhea in both sexes (LDT)).

No other clear treatment

- related eff ects were observed in the study.

Special Study

28

- d ay Subchronic Mucosal Irritancy Study of Three Sanguinarine Chloride Preparations in Dogs 46273802 (1987)
Acceptable/non

guideline This is a special study and no DER was prepared. However, a screening of the study findings did not impact the dermal or inhalation endpoints for the qualitative screen.

Special Study
Gingival Irritation Study in Dogs:

10% S anguinarine Chloride (57% NMP), 33% PLA Polymer, 50% Sanguinarine Chloride (35% NMP), 15% PLA Polymer, 100% Sanguinarine Chloride 46273805 (1990)
Acceptable/non

guideline This is a special study and no DER was prepared. However, a screening of the study findings did not impact the dermal or inhalation endpoints for the qualitative screen.

### Macleaya Extract/069095 Human Health Risk Assessment D418970

Page 14 of 17 Table A. 2.4 Subchronic, Chronic and Other Toxicity Profile for Macleaya Extract

Guideline No. Study Type MRID No. (year)

Classification

Dose Levels Results

870.3700
Developm ental toxicity

- rat ( s anguinaria extract (33% sanguinarine chloride and

— 68% total benzophenanthridine alkaloid)) 44525111(1989)
Acceptable/nonguideline

0, 5,20, and 60 mg/kg/day by
gavage in 1% citric acid (10 mL/kg)

From literature

46273808 (1988) GD

15

- 20
46273810 (1990) GD 6

- 15 Maternal NOAEL : 20 mg/kg/day

Maternal LOAEL :60 mg/kg/ reduced body weight gain and clinical toxicity (rales)

. Developmental NOAEL : Greater than 60 mg/kg/day
(HDT)
Developmental LOAEL : Could not be established.
This study was r eported as a scientific investigation (open literature study); therefore, many of the elements required under OPPTS guidelines were not present. Nevertheless, this study does provide some data which may be used for regulatory purposes. The study report (MR ID 46273810) provided more details with same

conclusions.

870.3700b Developmental Toxicity in non

rodents (rabbit)

( s anguinaria extract (33% sanguinarine chloride and

- 68% total benzophenanthridine alkaloid)) 44525111(1989)
Acceptable/ nonguideline 0, 5, 15,25, 50, and 75 mg/kg/day by gavage in
1% citric acid (4 mL/kg), GD 6

- 18

From literature

46273807 (1988) Maternal NOAEL = 15 mg/kg/day
LOAEL = 25 mg/kg/day based on decreased maternal body weight gain. Developmental NOAEL = 25 mg/kg/day
LOAEL = 50 mg/kg/day based on decreased number of fetuses/litter and increased postimpl a ntation loss.
This study was reported as a scientific investigation (open literature study); therefore, many of the elements required under OPPTS guidelines were not present . Nevertheless, this study does provide some data which may be used for regulatory purposes. The most significant deficiency in this study is the low number of litters available for examination. The study only started with 16 animals per group. Without the individual animal data, the pregnancy rate cannot be determined. In addition, for some parameters, the standard deviations are very high. Again, without the individual animal data, it cannot be determined whether or not the high

standard deviations are du e to a few outliers. The study report (MRID 46273807) did not provide additional information on these questions.

870.3800

1

- Generation Reproduction

- rat ( s anguinaria extract (33% sanguinarine chloride and

— 68% total benzophenanthridine alkaloid)) 44525111(1989)
Acceptable/ nonguideline 0, 10, 30, or 100 mg/kg/day by

gavage in 1% citric acid (10 mL/kg)

From literature

46273809 (1988) Parental NOAEL: 10 mg/kg/day
Parental LOAEL: 30 mg/kg bw/day (clinical signs indicative of central nervous system toxicity) Offspring NOAEL: 30 mg/kg/day Offspring LOAEL: 100 mg/kg bw/day (decreased body weight of pups at birth and during lactation) Reproductive NOAEL: Greater than 100 mg/kg/day (HDT) Reproductive LOAEL: Could not be established.
This study was reported as a scientific investigation (open literature study); therefore, many of the elements required under OPPTS guidelines were not present. Nevertheless, this study does provide some data which may be used for regulatory purposes. Stud y report (MRID 46273809) provides additional detail.

### Macleaya Extract/069095 Human Health Risk Assessment D418970

Page 15 of 17 Table A. 2.4 Subchronic, Chronic and Other Toxicity Profile for Macleaya Extract

Guideline No. Study Type MRID No. (year)

Classification

Dose Levels Results

870.4300 Chronic toxicity/ carcinogenicity in rodents (rat)
Sanguinaria extract (purity not reported) 45400505 (1989)
Unacceptable/ guideline 0, 5, 20, or 60 mg/kg/day by gavage (10 mL/kg) NOAEL = 20 mg/kg/day
LOAEL = 60 mg/kg/day based on decreased overall body weight gain in females and increased salivation and rales in both sexes.
Not oncogenic under conditions of study.
Excessive mortality, including in the control groups, prevented the 104 g avage errors. For the chronic toxicity portion of the study, a significant number of guideline measurements were not performed. These included ophthalmology, clinical chemistry, urinalysis, and organ weight determinations. In addition, the purity of the

sa nguinaria extract was not reported. However, some of the data may be useful for regulatory purposes.

870.5300

CHO/HGPRT Mammalian Cell Forward Gene Mutation Assay: Sanguinarine c hloride. 46273811 (1985) Acceptable/Guideline Sanguinarine c hloride was cytotoxic at doses of 0.3

- 8.0 μg/mL (

- S9) and at doses ≥20 μg/mL (+S9). Sanguinarine c hloride was not mutagenic up to toxic dose of 0.3 μg/mL (

- S9). Sanguinarine c hloride produced reproducible statistically significant increase in the mutant frequencies of the cultures treated with 10 μg/mL (+S9).

870.5375 Mammalian spermatogonial

chromosomal aberration In Vitro Mammalian Cytogenetic Test Using Human Peripheral Lymphocytes: Sanguinarine c hloride 46273812 (1991) Acceptable/Guideline Sanguinarine c hloride was insoluble at 500 μg/mL and higher. It was tested at 0.63, 1.25, 5, 10 μg/mL (

- S9) and at 1.25, 2.5, 5.0, 10, 20 μg/mL(+S9). The test material was soluble at these doses. Cytotoxicity was noted at the two higher doses. No statistically signific ant increase in structural chromosome aberrations was observed in the absence or presence of metabolic activation, regardless of harvest time (20 or 44 hours harvest).

870.5550 Rat Hepatocyte Primary Culture/DNA Repair Test: Sanguinarine c hloride. 46273813 (1985) Acceptable/Guideline Sanguinarine c hloride dissolved in DMSO was tested at 2, 5, 10, 20, 50, 100, 200, 400, 1000, 2000 μg/well. Dose levels at 200 μg/well and above produced net nuclear grain counts that were substantially greater than the solvent control. A dose related increases in net nuclear grains was observed at dose levels of 100, 200 and 400 /well reaching 3.4 net nuclear grains at the 400 μg/well dose. In addition, 30% of the cells had ≥ 5 net nuclear grains. A mean net nuclear gr ain of 5 or greater was not observed at any concentration. The results were considered to be equivocal.

870.5100

Bacterial reverse mutation test

Salmonella/Mammalian

- Microsome Plate Incorporation Mutagenicity Assay (Ames Test) And Escherichia

Coli WP2uvrA Reverse Mutation Assay: Sanguinaria e xtract 46273814 (1986) Acceptable/Guideline Sanguinaria e xtract was not posi tive in the E. coli Wp2uvrA with

or without aroclor

- induced rat liver microsomal activation. Sanguinaria e xtract was positive in the Salmonella strains TA98, TA100, TA1537, TA1538 (+S9). Sanguinaria e xtract was not mutagenic in Salmonella strains TA98,

TA1 00, TA1535, TA1537, TA1538 (

- S9) and TA1535 (+S9).

870.5385

Mammalian bone marrow

chromosomal aberration Micronucleus Cytogenetic Assay in Mice: Sanguinaria e xtract 46273815 (1987) Acceptable/Guideline IP injection of s anguinaria e xtract at 1.4, 7.1 or 14.2 mg/kg twice

with 24 hour interval between dosing, did not induce micronucleated polychromatic erythrocytes in male or female CD

1 mice. It was considered to be negative in this study.

### Macleaya Extract/069095 Human Health Risk Assessment D418970

Page 16 of 17 Table A. 2.4 Subchronic, Chronic and Other Toxicity Profile for Macleaya Extract

Guideline No. Study Type MRID No. (year)

Classification

Dose Levels Results

870.5395 Mammalian erythrocyte

micronucleus Unscheduled DNA Synthesis in Rat Primary Hepatocytes: Sanguinaria e xtract 46273816 (1987) Acceptable/Guideline Sanguinaria e xtract was tested at 11 dose levels ranging from 0.001 μg/mL to 100 μg/mL and was fully evaluated at 10.0, 3.0, 1.0, 0.3, and 0.1 μg/mL. It did not produce a significant increase in the mean number of net nuclear grain counts. It was considered to be negative in this test.

### Macleaya Extract/069095 Human Health Risk Assessment D418970

Page 17 of 17 APPENDIX B

Summary of Points of Departure Used in Risk Assessment

Toxicological doses/endpoints selected for the m acl e aya extract risk assessment are provided in

Table B.1

.

Table B.1 . Summary of Toxicological Doses and Endpoints for Macleaya Extract for Use in Occupational

Human Health Risk Assessments

Exposure/
Scenario Point of

Departure Uncertainty/

FQPA Safety

Factors RfD, PAD, LOC

for Risk
Assessment Study and Toxicological Effects

Acute Dietary No dietary exposure

Chronic Dietary No dietary exposure

Incidental Oral No dietary exposure

Dermal a
Short

- (1

- 30 days) and

Intermediate

- (1

- 6 months)

Term LOAEL =

10 mg/kg/day (LDT) ( NOAEL

not
established ) UF A = 10X

UF H = 10X

UF L = 3X Occupational

LOC for MOE

= 300 90

- day monkey study

( MRID 45400504 )
LOAEL = 10 mg/kg/day (LDT)

based on increased incidence (number of affected animals) and frequency of emesis and diarrhea

in both sexes

Inhalation b

Short

- (1

- 30 days) and

Intermediate

- (1

- 6 months)

Term LOAEL =

10 mg/kg/day (LDT) (NOAEL

not established) UF A = 10X

UF H = 10X

UF L = 3X Occupational

LOC for MOE =

300 90

- day monkey study

( MRID 45400504 )
LOAEL = 10 mg/kg/day (LDT)

based on increased incidence (number of affected animals) and frequency of emesis and diarrhea

in both sexes

Cancer (all routes) Carcinogenicity was not evaluated.

Point of Departure (POD) = A data point or an estimated point that is derived from observed dose

response data and used to mark the beginning of extrapolation to determine risk associated with lower environmentally rel evant human exposures. NOAEL = no observed adverse effect level. LOAEL = lowest observed adverse effect level. UF = uncertainty factor. UF A = extrapolation from animal to human (interspecies). UF H = potential variation in sensitivity among members of the human population (intraspecies). UF L = use of a LOAEL to extrapolate a NOAEL. UF S = use of a short

- term study for long

term risk assessment. UF DB = to account for the absence of key date (i.e., lack of a critical study). FQPA SF = FQPA Safety Facto r. PAD = population adjusted dose (a = acute, c = chronic). RfD = reference dose. MOE = margin of exposure. LOC = level of concern. N/A = not applicable.

a 10% default dermal absorption factor is used.

b Since no inhalation absorption data are available, toxicity by the inhalation route is considered to be equivalent to the estimated toxicity by the oral route of exposure (100% absorption factor).