SRC TR-10-056

HPV1 TEST RULE – DATA ADEQUACY ASSESSMENT

Methanesulfinic acid, hydroxy-, mo nosodium salt, CAS No. 149-44-0

Repeated-Dose with Reproductive/D evelopmental Toxicity Screen

Prepared by:

Teresa Manyin, Chris Janson, and Joan D. Garey

Environmental Science Center SRC, Inc.

7502 Round Pond Road North Syracuse, NY 13212

Contract No. EP-W-09-027

Task FG007.0.009.004

Submitted to:

OPPT/RAD/HPVCB (7403M) U.S. Environmental Protection Agency 1200 Pennsylvania Avenue

Washington, D.C. 20460

Elizabeth Margosches, Project Officer Meena Sonawane, Work Assignment Manager David Brooks, Alternate Wo rk Assignment Manager

March 5, 2010HPV1 Test Rule – Data Adequacy Assessment

Methanesulfinic acid, hydroxy-, mo nosodium salt, CAS No. 149-44-0

Repeated Dose with Reproductive /Developmental Toxicity Screen

March 2010

Submitted to: High Production Volume Chemicals Branch

Risk Assessment Division

Office of Pollution Prevention and Toxics Environmental Protection Agency

WAM: Meena Sonawane

Alternate WAM: David Brooks 1200 Pennsylvania Avenue, NW

Washington, DC 20460-0001

Prepared By:

SRC, Inc.

Environmental Science Center 2451 Crystal Drive, Suite 804 Arlington, VA 22202Draft HPV1 Test Rule – Data Adequacy – Repeated-Dose/Repro/Develop Tox CAS No. 149-44-0

PREPARATION OF THE DAT A ADEQUACY ASSESSMENT

This document contains the review of a toxi city study conducted in response to the first

HPV TSCA Section 4 Test Rule (71 FR 13708, March 16, 2006). Using the applicable test guideline and professional judgment as guide s, the study was reviewed for adequacy and evaluated for compliance with the test guideline. (T he applicable test guid eline is not always the test guideline required by the test rule, because some test sponsors requested and received approval from EPA to use a different guideline .) The study was summarized according to a standard format, assessed for adequacy, and assi gned a Klimisch reliability code. If the study was determined to be inadequate, a basis for th at conclusion is supplie d. For studies already assigned Klimisch codes, an explanation is prov ided if the reviewer’s assessment of data adequacy differed from the pre-assigned Klimisch code.

A detailed evaluation of the test report a nd study conduct is provided at the end of the summary document in a section entitled “Revie w of TSCA Section 4 Final Study Report”. Any deviations from the applicable test guideline are described. The study is also evaluated for compliance with Standard Operating Procedur es, Good Laboratory Prac tice Standards, and Quality Assurance procedures, as well as the use of appropriate controls and test substances. Any deficiencies in the descri ptions of the study method, test system, animals studied, physical conditions, dosage information, indi vidual animal data, calculations, and statistical analyses are described, and internal inconsistencies within the study report are identified. An overall assessment is provided for each of the following: study conduct, data reliability, and data utility.

1Draft HPV1 Test Rule – Data Adequacy – Repeated-Dose/Repro/DevelopTox CAS No. 149-44-0

REPEATED-DOSE/REPRODUCTIVE/ DEVELOPMENTAL TOXICITY

Title: A combined 28-day repeated dose toxicity study with the reprodu ction/developmental toxicity screening test of methane sulfinic acid, hydroxy- , monosodium salt in rats by oral

gavage

The study is: Adequate Not Adequate

TEST SUBSTANCE

Identity: Methanesulfinic acid, hydroxy-, monosodi um salt, aka sodium hydroxymethane sulfinate (SFS), CAS No. 149-44-0

Remarks: 88.4% purity by weight (7 6.9% sodium hydroxymethane sulfinate, 11.5% water); 100.3% if calculated as dihydrate; impurities and stabi lizers include 1.9% alka linity (as sodium carbonate), 4.4% sodium hydroxymethanesulfonate, 1.9% sodium sulfite, 2.4% sodium sulfate, 0.2% iron (as Fe), and 0.2% zinc (as Zn); Batch Number 07041205 from Brüggemann; commercial substance; white powder of a crystall ine substance consisting of colorless crystals;

MW = 118.08

METHOD

Method/guideline followed: OECD 422 and EPA, 40 CFR 799.9365

Test type: Combined 28-day repeated-dose toxicity study with reproductive/developmental toxicity screening test

GLP compliant?: Yes

Year study performed: 2007

Test species: Male and virgin female Wistar (Crl:WI(H an); outbred, SPF-Quality) rats; supplier: Charles River, Deutschland, Sulzfeld, Germany; age at initiation: appr oximately 10 weeks old; mean initial body weights: males: 292 g, females: 190 g

Number of animals/sex/dose: 10/sex/dose

Route of administration: Oral gavage

Vehicle: Water (Elix, Millipore S.A.S., Molsheim, France)

Exposure duration: Males were exposed for 30 days (dur ing 2 weeks prior to mating, during mating [up to 14 days], and to termination). Females were exposed for 42 - 53 days (during 2 weeks prior to mating, during mating and ge station, and up to Day 4 of lactation).

Doses/concentrations tested: 0, 100, 300, and 1000 mg/kg-bw/day

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Statistical methods used: Normally distributed data were an alyzed using the Dunnett test, based on a pooled variance estimate. The Steel test was applied if data were not normally distributed. The Fisher Exact test was applied to frequenc y data. The following were calculated for each group: percentage mating, fertility index, concepti on rate, gestation index, duration of gestation, percentage of live males at first litter check, percentage of live females at first litter check, percentage of postnatal loss from Days 0-4 post partum, and viability index. All tests were twosided, with p < 0.05 accepted as the lowest level of significance.

Test method/conditions remarks: Animals were acclimated at least 5 days prior to start of treatment. Th ey were housed in rooms maintained at 19.0-23.5 °C, with a relative humid ity of 26-95% and a 12-hour light/ 12 hour dark cycle. There were 15 air changes per hour. Prio r to mating, animals were housed in groups of 5 animals/sex/cage in Macrolon cages (MIV type, he ight 18 cm). Females were caged with males on a one-to-one basis during mating. After mating, males were housed in their home cage, with a maximum of 5 animals/sex/cage. Females we re individually housed after mating and were kept with their offspring until termination.

Cages were generally supplied with sterilized sawdust as beddi ng material and paper as cageenrichment. During overnight activity monito ring, animals were hous ed individually with

sterilized sawdust beddi ng material without paper cage-enrich ment. Animals were allowed free access to tap water and pelleted rodent diet (SMR/M-Z from SSNIFF Spezialdiaten GmbH, Soest, Germany). Analyses of bedding, paper, diet and water did not reveal any findings that were considered to affect study integrity.

Test material: Formulations (w/w) were prep ared daily, within 4 hours of dosing, and were homogenized to a visually acceptable level. No adjustment was made for specific gravity, vehicle and/or formulation. The pH of the solu tions was determined weekly. The pH of a 10% solution was approximately 10. Sampling and chemi cal analysis of formul ations were performed on one day during the treatment period; duplicate samples from each group were analyzed for accuracy. Duplicate samples of dosing solutions taken from the bottom, middle and top for the low- and high- dose groups were analyzed for homogeneity and stability at 0 hours after preparation. Duplicate samples of these solutions (taken from the middle) were also analyzed for stability at 5 hours after prepara tion. The analytical method used was based on the results of a separate project for the development and validat ion of the analytical method (NOTOX Project 485209).

The pHs of the formulations were be tween 8.31-8.68 (control), 10.12-10.17 (low dose), 10.10- 10.17 (mid dose) and 9.94-10.02 (high dose). The accuracy of the formulations was 102-109% (low dose), 98-103% (mid dose) and 87-105% (high dose). Based on the results, it was concluded that the formulations were prepared correctly. The low- and high-dose formulations were homogeneous ( ≤ 10% coefficient of variation). Form ulations at the entire range were

stable for at least 5 hours when stored at room temperature.

A health examination was performed prior to treat ment to ensure that the animals were in a good

state of health. A total of 40 animals/sex were randomized into four groups of 10/sex according

3Draft HPV1 Test Rule – Data Adequacy – Repeated-Dose/Repro/DevelopTox CAS No. 149-44-0

to body weight, with all animals w ithin 20% of the sex mean. They were identified by earmark

and tattoo.

Groups of animals were given 0 (water vehicle), 100, 300 or 1000 mg/kg-bw test material (at 5 mL/kg-bw) by oral gavage, once daily, 7 days/week up to the day of scheduled termination. A limit dose of 1000 mg/kg-bw/day was selected as the high dose level based on data obtained in a preliminary range-finding study. Actual dose volu mes were calculated according to the latest body weight. Following 14 days of exposure, males and females were co-housed (1:1). Sibling mating was avoided. Detection of mating was co nfirmed by presence of sperm in the vaginal lavage or by the appearance of an intravaginal copulatory plug. This da y was designated as Day 0 postcoitum. Once mating had occurred, the male s and females were separated. A maximum of 14 days was allowed for mating. There were at le ast 8 pregnant females per dose level. Females were allowed to litter normally. Day 1 of lact ation was defined as the day when a litter was found completed. Females that we re littering were left undisturbe d. On Day 1 of lactation, all pups were individually identified by a tattoo. Examination of maternal care revealed no

deficiencies.

Parental animals were observed for mortality and viability at least twi ce daily. At least once daily, detailed clinical exams were conducted for a ll animals. This was also performed outside the home cage once before treatment and at week ly intervals. These observations were not performed when the animals were mating or housed individually. The time of onset, degree and duration was recorded. Cage debr is of pregnant females was ex amined for evidence of abortion or premature birth. No signs of abortion, prem ature birth or difficult or prolonged parturition

were recorded.

Functional observations (hearing ab ility, pupillary reflex, static righ ting reflex and grip strength and a motor activity test) were recorded for the first five mated males/group and the first five females/group with live offspring. During the motor activity test, males were caged individually and females were caged with their offspring. Th e males were tested during week 4 of treatment and the assigned females were tested during lactation (all before blood sampling).

Males and females were weighed on the first da y of exposure and weekly thereafter. Mated females were weighed on Days 0, 4, 7, 11, 14, 17 and 20 of gestation, and during lactation on Days 1 and 4. Food consumption was recorded weekly (with the ex ception that it was not recorded during breeding). Food consumption of mated females was measured on Days 0, 4, 7, 11, 14, 17 and 20 post-coitum and after delivery on Days 1 and 4 postpartum. Water consumption was not quantified, but was monitored subjectively. Identities of animals paired, mating date, confirmation of pregnanc y, and delivery day were recorded.

After an overnight fast, blood was collected from th e retro-orbital sinuses of the first five mated males/group and the first five females/group with live offspring, under isoflurane anesthesia. Standard hematological (white blood cells, di fferential leukocyte count, red blood cells, reticulocytes, red blood cell distribution widt h, hemoglobin, hematocrit, mean corpuscular volume, mean corpuscular hemoglobin, mean cor puscular hemoglobin conc entration, platelets, prothrombin time, and activated partial throm boplastin time) and c linical biochemistry parameters (alanine aminotransferase, aspartat e aminotransferase, alkaline phosphatase, total

4Draft HPV1 Test Rule – Data Adequacy – Repeated-Dose/Repro/DevelopTox CAS No. 149-44-0

protein, albumin, total bilirubin, urea, creatinin e, glucose, cholesterol, sodium, potassium, chloride, calcium, and inorganic phosphate) were performed after appropriate blood processing.

On lactation Day 5 (or shortly ther eafter), parental females were an esthetized with isoflurane and exsanguinated. Animals were subjected to a gros s examination, organ weights (adrenal glands, brain, heart, kidneys, liver, spl een, and thymus) of the first 5 females with live offspring per group were collected and terminal body weights were recorded. Ti ssues (adrenal glands, aorta, brain, cecum, cervix, clitoral gland, colon, duodenum, eyes with optic nerve and Harderian gland, mammary gland area, femur, heart, ileum , jejunum, kidneys, larynx, exorbital lacrimal gland, liver, lung, mandibular and mesenteric lymph nodes, nasopharynx, esophagus, ovaries, pancreas, Peyer's patches, pituitary gland, pr eputial gland, rectum, mandibular and sublingual salivary glands, sciatic nerve, skeletal muscle, skin, spinal cord, spleen, sternum with bone marrow, stomach, thymus, thyroid with parathyr oid, tongue, trachea, and urinary bladder) and gross lesions of these animals were preserved fo r possible future histopathological examination. The cervix, clitoral gland, ovaries , preputial gland, uterus, vaginaand all gross lesions from remaining animals also were preserved. Slides of preserved organs a nd tissues of selected control and high dose animals and gross lesions of all animals were prepared and examined by a pathologist. The numbers of former implanta tion sites and corpora lu tea were counted and

recorded.

On post-mating Days 24-26, females that had not delivered were anesthetized and exsanguinated. No organs were collected for weighing. Animal s were subjected to a gross examination and tissues described above for parental females (and any gross lesions) were preserved. The reproductive organs of all females that failed to conceive were examined by a pathologist. Additionally, nongravid uteri were stained using the Salewski t echnique to identify former implantation sites.

After completion of the mating period, parental males were anesthetized with isoflurane and exsanguinated and grossly examined. Organ weights (adrenal glands, brain, heart, kidneys, liver, spleen, and thymus) of the first 5 mated male s per group were collect ed and terminal body weights were recorded. Weights of epididymides and testes of all males were also recorded. The same tissues preserved for selected females (see above) were preserved for the first 5 mated males per group (with the exception of female reproductive organs and the addition of the epididymides, prostate gland, seminal vesicles, te stes, and coagulation gl and). The coagulation gland, epididymides, preputial glan d, prostate gland, seminal vesicles , testes and a ll gross lesions from remaining animals also were preserved. Slid es of preserved organs and tissues of selected control and high-dose animals and gross lesions of all animals were prepared and examined by a pathologist. Of the selected 5 males/group of the control and high-dose group, additional slides of the testes were prepared to examine staging of spermatogene sis. The reproductive organs of all males that failed to sire were examined by a pathologist.

The numbers of live and dead pups at the first litter check (Day 1 of lactation) and daily

thereafter were determined. Detailed clinical ob servations of pups were made at least once daily. Live pups were weighed during l actation Days 1 and 4. Sex of all pups was determined on Days 1 and 4 of lactation (by measur ing the anogenital distance).

5Draft HPV1 Test Rule – Data Adequacy – Repeated-Dose/Repro/DevelopTox CAS No. 149-44-0

Pups were killed by decapitation on Day 5 of lactation (or shortly thereafter). All offspring were sexed and externally examined. Stomachs were ex amined for the presence of milk. Descriptions

of all external abnormalities were recorded. If possible, defects or causes of death were evaluated. Any abnormal pup, organ or tissue wa s preserved for possible future examination.

RESULTS

Endpoint value(s): The parental LOAEL was 1000 mg/kg- bw/day, based on reductions in mean body weight, body weight gain, and food consumpti on, as well as increase s in the number of reticulocytes (males only) and the level of alanin e aminotransferase (males only). The parental NOAEL was 300 mg/kg-bw/day. The reproductive and developmental NOAEL was 1000 mg/kg-bw/day, based on no adverse effect s observed at the highest dose tested.

Results remarks: Formulation analysis showed that the formulations were prepared accurately, were homogeneous (i.e., coefficient of variation ≤ 10%) and were stable for at leas t 5 hours at room temperature. The accuracy of formulation (determined by chemi cal analysis) for the low-, mid-, and high-dose groups were 102-109%, 98-103%, and 87-105%, respectively.

Parental animals (effects by dose level): 1000 mg/kg-bw/day: None of the animals died a nd functional observations were not affected. One female showed hunched posture and piloer ection from Day 20 post-coitum until Day 3 of

lactation. Piloerection was noted in another female on Day 4 of lactation. Slight salivation noted incidentally in both sexes was attributed to the taste of the formulation and was not considered toxicologically relevant. Lower mean body wei ghts were recorded for males and females at 1000 mg/kg-bw during the complete treatment period (n ot always statistically significant). Body weight gains of males were also lower than the control throug hout the treatment period. For females, body weight gain was decreased on Day 8 during the pre-mating pe riod and on Day 1 of mating. Food consumption before or after allo wance for body weight was reduced in males and females during the whole observation period (not al ways statistically significant). A slightly higher white blood cell count was noted in males (not statistically significant) and females. Relative numbers of reticulocyte s were increased in males (3.6 ± 0.8% in treated vs. 2.3 ± 0.1 % in control). Increased mean corpuscular volume and decrease d mean corpuscular hemoglobin

concentration in females were not co nsidered to be related to treatm ent as they were either within the range of normal or showed no dose-re sponse relationship. Increased alanine aminotransferase was found in males (59.2 ± 20.4 U/I in treated vs. 38.0 ± 3.6 U/I in control). One male had increased activity of aspartate am inotransferase. Increased total bilirubin and

decreased sodium in males were not considered to be related to treatment due to the lack of a dose-response relationship. One male showed exopht halmus of the eye, which was considered to be caused by blood sampling. Increases in orga n/body weight ratios for the liver (2.57 ± 0.14% in treated vs. 2.33 ± 0.12% in control), spl een (0.225 ± 0.032 in treated vs. 0.162 ± 0.033 in control), testes (1.16 ± 0.15% in treated vs. 0.94 ± 0.16% in control) and epididymides (0.388 ± 0.071 % in treated vs. 0.301 ± 0.050 in control) of males and liver (3.64 ± 0.22% in treated vs. 3.04 ± 0.33% in control), spleen (0.287 ± 0.023% in treated vs. 0.225 ± 0.0303% in control), kidneys (0.83 ± 0.05% in treated vs. 0 .73 ± 0.06% in control) and adrenals (0.043 ± 0.004% in treated vs. 0.034 ± 0.004% in control) of fema les were observed. There were no corresponding

6Draft HPV1 Test Rule – Data Adequacy – Repeated-Dose/Repro/DevelopTox CAS No. 149-44-0

histological changes. An incr ease in the organ/body weight ra tio of the brain in males (0.66 ± 0.05% in treated vs. 0.60 ± 0 .01 % in control) was considered to be due to the relatively low terminal body weight (300 ± 14 g in treated vs. 336 ± 19 g in control). Absolute brain weight (but not relative) was slightly decreased in females (1.77 ± 0.05 g in treated vs. 1.87 ± 0.05 in control). Reproductive and breeding parameters were not affected by treatment at any dose tested. No treatment-related findings were observe d for mating performance, fertility parameters (fertility index, conception rate, and gest ation index), and duration of gestation.

300 mg/kg-bw/day: None of the animals died a nd functional observations were not affected. Females had slightly lower mean absolute body weights on Day 8 of premating (189 ± 6.6 g in treated vs. 197 ± 7.2 g in control). As this had recovered on Day 1 of mating (198 ± 11.8 g in treated vs. 203 ± 8.2 g in control), it was not considered to be toxi cologically relevant. Increased basophils in males, increased mean corpuscula r volume and mean corpuscular hemoglobin in females, decreased total bilirubin in males, and decreased albumin in females were not considered to be related to treatment as they we re either within the rang e of normal or showed no dose-response relationship. No ot her abnormal findings were noted.

100 mg/kg-bw/day: None of the animals died a nd functional observations were not affected. Females had slightly lower mean absolute body weights on Day 8 of premating (189 ± 6.6 g in treated vs. 197 ± 7.2 g in control). As this had recovered on Day 1 of mating (196 ± 6.3 g in treated vs. 203 ± 8.2 g in control), it was not considered to be toxi cologically relevant. Increased basophils in males and decreased total bilirubin in males were not considered to be related to treatment as they were either within the range of normal or showed no dose-response relationship. No other abnormal findings were noted.

Control: One female showed exophthalmus of the eye, which was considered to be caused by blood sampling.

Offspring: The numbers of dead and living pups at first litter check were not affected by exposure to the test substance. Postnatal loss and viability index were similar for the control and treated groups. Development of pups was not affected by trea tment with up to 1000 mg/kg-bw/day. Mean pup body weights were similar for the control and treated groups (means ranged from 5.6-5.8 g and 5.3-5.6 g in males and females on Day 1, and 8.3-8.8 and 8.0-8.6 g in males and females on Day 4, with none of the low values in the listed ranges being in high -dose animals). None of the clinical or macroscopic findings (scabs at the nec k, dark red discoloration of tail apex, blue spot on neck, wound at the head) were co nsidered to be related to trea tment as they were considered to be within normal biological variation. Summary Table: Parameter Control 100 mg/kg 300 mg/kg 1000 mg/kg Percentage Mating 100 100 100 100 Fertility Index 80 100 90 100

Gestation Index 100 100 100 100

Mean Precoital Time* 2.5 2.9 1.4 1.7

Duration of Gestation (days) 21.1 ± 0.4 21.1 ± 0.3 21.0 ± 0.0 21.1 ± 0.3 Dead Pups at

7Draft HPV1 Test Rule – Data Adequacy – Repeated-Dose/Repro/DevelopTox CAS No. 149-44-0

First Litter Check 0 0 0 0 Postnatal Loss (%) 0.0 2.6 2.9 1.0

Litters Affected 0 3 2 1 Viability Index 100.0 97.4 97.1 99.0 *units not listed, but presumed to be days

CONCLUSIONS

Treatment with methane sulfinic acid, hydroxy-, monosodium salt by oral gavage in male and female Wistar Han rats at dose levels of 100, 300 and 1000 mg/kg-bw/day revealed parental toxicity at 1000 mg/kg-bw/day. No reproduc tion, breeding, and developmental toxicity was observed for treatment up to 1000 mg/kg-bw/day. Based on these findings, the parental NOAEL was established at 300 mg/kg-bw/day. The defi nitive reproduction, breeding, and developmental

NOAEL was established as being 1000 mg/kg-bw/day.

STUDY RELIABILITY

[1] = Reliable without restriction.

REFERENCE

Beekhuijzen, M.E.W. 2008. A combined 28-da y repeated dose toxicity study with the reproduction/developmental toxicity screeni ng test of methane sulfinic acid, hydroxy-, monosodium salt in rats by oral gavage. NOTOX B.V. Project No. 485218 (and 485615), dated

12 June 2008

8Review of TSCA Section 4 Final Study Report

CITB Project Manager: Catherine Roman

Chemical Name: Methanesulfinic acid, hydroxy-, monosodium salt

CAS Registry Number: 149-44-0

Report Title: A combined 28-day repeated dose toxic ity study with the reproduction/developmental toxicity screening test of methane sulfinic acid, hydroxy-, monosodium salt in rats by oral gavage

Test Sponsor: The SFS Consortium, consisting of BrüggemannChemical US, Inc. and Royce Associates

Testing Lab: NOTOX B.V., Hambakenwetering 7, 5231 DD 's-Hertogenbosch, The Netherlands

Test Standard: 40 CFR 799.9365

Date Study Received by EPA: June 2008 Date Study Due to EPA: May 17, 2007

Special Instructions (e.g., regarding approved protocol modifications, etc.):

Part I. Test Report and Study Conduct

Enter one of the following codes into each of the checkboxes 1 through 13: Agree [ A ], Disagree [ D ], Not Applicable [ NA ], or Cannot Determine [ C ], and add comment, below, as needed

[D] 1. The final study report indicates evidence of gross, unauthorized or undocumented deviation from the EPA test standard requirements, including “shall,” “must,” or “will” violations. List significant test standard deviations and “shall,” “must,” or “will” violations in comments below.

[D] 2. The final study report indicates evidence of deviations from the “should” statements in the study protocol that may have a bearing on the study conclusions. List these in comments below.

[D] 3. Approved modifications of test standard or testing protocol (CITB PM describe or attach) were not followed or are inadequately documented.

[D] 4. There is evidence that laboratory Standard Operating Procedures (SOPs) were inadequately followed.

[D] 5. The Statement of Quality Assurance (QA) inspecti ons, their dates, and findings were missing or not signed. There is evidence that QA pr ocedures may be inadequate.

[D] 6. There is evidence that Good Laboratory Practice Standards (GLPS) were inadequately followed.

[D] 7. The test, control, or reference substances are di fferent from that specified in the section 4 test rule or

Enforceable Consent Agreement (ECA).

[D] 8. Animal identification is inadequate or unclear; use of unhealthy animals or other factors (e.g., deviations from temperature, lighting, dosing) are apparent.

[D] 9. The final study report inadequately describes the study method.

[D] 10. The final study report inadequately describes the test system.

[D] 11. The final study report inadequately describes the dosage information.[D] 12. The final study report does not accurately reflect the individual animal data (or that of other discrete observations pertinent to the type of study conducted), or individual data were not provided.

[D] 13. The final study report inadequately describes the calculations, interpolations, data transformations, statistical analyses, or modeling parameters used.

Part II. Conclusions

Place a check mark beside the summar y statement(s) that apply to this review and add comments below as appropriate

Part A. To be filled out by Reviewer

Study Conduct 1. The study was well-conducted. [X] 2. The study was generally well-conducted, with some minor [ ] deficiencies (list in notes below). 3. The study conduct was significantly deficient or questionable, (give [ ] reasons in notes below).

Data Reliability 4. The study results are precise and accurate. [X] 5. Appropriate methods were used to determine [X] statistical significance. 6. If repeated, the test results would be expected to be the [X]

same.

Data Utility 7. The data are useful for ri sk assessment for the endpoint tested. [X] 8. The data are useful for some other purpose (e.g., dose-setting). [ ] 9. The data are not useful for any purpose (describe in notes below). [ ]

Reviewer Comments (continue on separate page, if needed):

Reviewer Name/Title/Br anch/Division/Office: Teresa Manyin/SRC, Inc.

Reviewer Signature: David Brooks/HPVCB/RAD/OPPTS

Part B. To be filled out by CITB PM

Tiered Testing Decision 1. The tiered testing trigger was me t, additional testing is indicated. [ ] (when applicable) 2. The trigger for additional (tiered) testing was not met. [ ]

Study Acceptability 3. The study is acceptable. [ ] 4. The study may not be acceptable [ ] (provide rationale in memo to CITB chief).