— Sponsor ee Bil Liao NELSOI Jingzhou Haixin Green Cross Med. Prod. Co., Ltd Century Bidg. 19F A, Jianghan RD 206 Wuhan City, Hubei CHINA CN 430010 ISO 10993 Part 10 Guinea Pig Buehler Sensitization Tes! LABORATORIES ‘Summary: Enclosed is the final report for the testing we coordinated for you. The information is retained by the testing laboratory. Test Article: ‘Surgical Mask Nelson Laboratory Number: 634772 Testing Lab: sinclair Research Results: The test article (device) extracts demonstrated no sensitization reactions under the conditions of this assay. It you have any questions, please feel free to call or email any of our Subcontracting personnel at 601- 290-7500 or subcontractina@nelsonlabs.com. Thank you for testing with Nelson Laboratories, Inc. 20 Wu 2o\2. Mindy Schvaneveldt, AS. Date© sinclair research STUDY TITLE: PROTOCOL NUMBER: STUDY NUMBER: TEST ARTICLE NAME: TEST ARTICLE LOT NO: TEST FACILITY: SPONSOR: NELSON REFERENCE NO: DATA REQUIREMENTS: DATE AMPLE RECEIVED: STUDY INITIATION DATE: STUDY COMPLETION DATE: RESULTS SUMMARY: (AceREDITED) Dog et or FINAL REPORT ISO 10993 Part 10 Guinea Pig Buehler Sensitization Test D10971.046 (Version 2) D10971.046-28 Surgical Mask NA Sinclair Research Center (SRC), LLC. (AAALAC Accredited) 562 State Road DD ‘Auxvasse, MO 65231, USA Phone: (573) 387-4400 Fax: (573) 387-4404 Nelson Laboratories, Inc. 6280 South Redwood Road Salt Lake City, UT 84123 634772 GLP 23-May-2012 23-May-2012 18-Jul-2012 The test article (device) extracts demonstrated no sensitization reactions under the conditions of this assay.‘SRC Study No, Di Final 0 QUALITY ASSURANCE UNIT SUMMARY ‘The study has been performed under Good Laboratory Practice regulations (FDA, 21 CFR, Part 58 - Good Laboratory Practice for Non-clinical Laboratory Studies) and in accordance with standard operating procedures and study protocol. The quality assurance unit inspected this study on the dates listed below. The report accurately reflects the raw data. Date Reported to Study Director Phase Inspected Date Inspected ‘& Management Protocol 10-Feb-2012 10-Feb-2012 48 hour Skin Score 29-Jun-2012 29-Jun-2012 Draft Report and Data 05-Jul-2012 05-Jul-2012 Final Report 18-Jul-2012 18-Jul-2012 Ashley Abernathy GOOD LABORATORY PRACTICES STATEMENT ‘The SRC study referenced in this report was conducted in compliance with Good Laboratory Practice (GLP) Regulations set forth in Title 21 Part 58 of the Code of Federal Regulations of the United States of America. Other portions of this study that were not performed by or under the direction of SRC, including the characterization and stability testing of the test le (device), were the responsibility of the sponsor and are exempt from this GLP statement. Study Director: KEY STUDY PERSONNEL Luke Zhang, M.D., M.Se. Study Director Jason Liu, Ph.D. Test Facility Management Chris Hanks, DVM, M.S. Staff Veterinarian Dean Wehmeier, B.S. Animal Facility Management Beth Derendinger, B.S. Data QC and Report Writing‘SRC Study No, D10971,046-28 Ei Page 3 of I OBJECTIVE ‘The objective of this study was to assess the potential of the test article (device) to produce a dermal sensitization reaction using the ISO Buehler Closed Patch Sensitization Test as required of medical device biocompatibility as identified in ISO 10993 Part 10. TEST ARTICLE IDENTIFICATION Test Article Name: Surgical Mask LovBatch #: NA Nelson Reference No.: 634772 SRC Test Article ID #: 3832 Sterile/ Non-sterile: Non-sterile Storage Conditions: Room temperature Disposition: Al used and unused discarded at the completion Description: White surgical mask with ear loops MSDS/CofA: NA ‘The test article was cut prior to dose. One piece of the surgical mask (part of mask with metal strip not used due to inability to cut material) and ear loops were included on ‘each patch for the test. Patch size: ~2 »3 em Preparation: CONTROL ARTICLE IDENTIFICATION AND PREPARATION Control Article Name: Cotton Gauze Supplier: ‘TW Medical LoBatch #: ABI911 Expiry: NA Preparation: The control article was cut prior to dose. Patch size: ~2 »3 cm TEST ARTICLE CHARACTERIZATION The sponsor was responsible for all test article characterization data as specified in the GLP regulations. The identity, strength, stability, purity, and chemical composition of the test le was solely the responsibility of the sponsor. It was the responsibility of the sponsor to ensure that the test article submitted for testing was representative of the final product that ‘was subjected to materials characterization, EXPERIMENTAL DESIGN Seventeen (17) adult Guinea pigs were used in this study with 11 animals in the test group and 6 animals in the control group. Study day 0 was the first day of dosing. The test article‘SRC Study No, D10971,046-28 Final Report Page 4 of 10 and control article were dosed in two phases (induction and challenge) over the proposed duration. Dermal scoring was performed at 24 + 2 and 48 + 2 hours post challenge phase patch removal. All animals were euthanized at the end of the study, JUSTIFICATION FOR SELECTION OF THE TEST SYSTEM ‘The Guinea pig is recognized as the standard model to predict dermal sensitization reaction in humans, and is recommended by the FDA for dermal sensitization studies. For this reason, the Guinea pig was used in this study to evaluate the potential dermal sensitization reaction of the test article. INSTITUTIONAL ANIMAL CARE AND USE COMMITTEE (IACUC) ‘The care and use of animals on this study were reviewed and approved by SRC IACUC. IACUC Protocol Approval Date: 7-Mar-2011 IDENTIFICATION OF TEST SYSTEM Species/Strain: Cavia porcellus, albino Guinea Pig Source: Elm Hill Labs (Chelmsford, MA) Age: 7.7 weeks ‘Weight Range: 442-513 g Gender: Female Identification Ear tag and cage card No. of Animals Used: Total: | 17 |Comtro:| 6 | Test: | Il In-life Duration (Termination): _ | Euthanized on Study day 35 HUSBANDRY Acclimation: Pre-acclimated Housi Animals were group housed (5-6 animals/cage). Room temperature and lighting were monitored and controlled while Environment: humidity was monitored, but not controlled. ‘Animals were provided ad libitum food and water daily. Food and water Food/Water: analyses are maintained as facility records and there were no known contaminants expected to interfere with the test results. ANIMAL SELECTION AND RANDOMIZATION Animals were selected for study based on a physical examination. The selected animals were randomized into treatment groups.‘SRC Study No. D10971,046-28 Final Report Page 5 of 10 ANIMAL PREPARATION ‘The hair on the target dose areas (~S x 7 cm on the left flank area for the right flank area for challenge phase) was clipped prior to dose adm ‘TEST AND CONTROL ARTICLE ADMINISTRATION Induction Phase (Day 0-20) A patch of control or test article of ~ 2 x 3 cm size was applied to the designated left dose site of control or test animal, respectively, at each dose administration. The dose area was ‘wrapped with an elastic bandage and secured with hypoallergenic tape. The patch and dressing were removed at 6 + 0.5 hours after application. This procedure was repeated three (3) times per week for three (3) consecutive weeks. Challenge Phase (Day 33-35) A patch of control or test article of ~ 2 » 3 cm size was applied to the designated dose site on the right sides of control or test animals, respectively, at each dose administration. The dose area was wrapped with an elastic bandage and secured with hypoallergenic tape. Each patch and dressing was removed at 6+ 0.5 hours after application. GENERAL OBSERVATIONS AND DOSING SITE SCORE All animals were observed daily for general appearance. Dermal observations of the challenge phase dose sites were performed at 24 + 2 and 48 + 2 hours after patch removal. Scores could range from 0 to 3 as shown in Table 1 ‘Table 1: Dermal Observation Scoring System \duction phase and on Reaction Score No visible change 0 Discrete or patchy erythema 1 Moderate and confluent erythema 2 Intense erythema and swelling (edema) 3 TERMINATION All animals were euthanized with Fatal Plus IP injection at the termination of the study. EVALUATION CRITERIA ‘Test results were based on incidence and severity of the sensitization reaction. Incidence was defined as the percentage of animals exhibiting a sensitization reaction at each observation time point (24 and 48 hours) post challenge. Severity was calculated by dividing the sum of the challenge scores by the total number of animals in each group at each time point. Severity scores of one (I) or greater in the test group generally indicated sensitization, provided grades of less than one (1) were observed on the control animals. If severity of one (1) or greater was noted on the controls, then the reaction of the test animals exceeded the most severe control reaction to be considered due to sensitization. In the final analysis of data, consideration was given to the overall patterns, intensity, duration, and character of reactions‘SRC Study No. D10971.046-28 Final Report Page 6 of 10 of the test as compared to those of the control animals. The skin reaction results are presented as positive (+, sensitization) or negative (-, non-sensitization).. ASSAY VALIDITY ‘The assay was considered valid based upon the fact that there was no excessive skin irritation in control animals and the test area consisted of normal skin prior to the test. The control article sites did not receive any dermal scores greater than zero. METHOD FOR CONTROL OF BIAS. Although the study was not blinded, the use of a control article controlled experimental bias. ARCHIVAL ‘A copy of the original report, original protocol (including amendments, deviations, Test Requisition Form and attachments) and all original in-life study specific raw data as well as pertinent in-life facility data are archived at Sinclair Research Center LLC. COMPLIANCE ‘The procedures including care, housing and handling of the animals were performed in compliance with the ISO 10993-10: Tests for Irritation and Skin Sensitization. The study was conducted and reported in compliance with U.S. Food and Drug Administration Good Laboratory Practice (GLP) regulations set forth in 21 CFR Part 58 and in accordance with the global protocol and the associated Test Requisition Form, facility standard operating procedures, and USDA Policy 12. Protocol amendments and deviations, if any, were handled according to facility SOP. POSITIVE CONTROL DATA Sinclair Research Center conducts positive control tests approximately every 6 months. The most recent positive control test was completed on 02 Apr 2012 (SRC study number: D10971.050-2). The methods used for the positive control assay were similar to the reported test. The same strain of animal was used. The sensitizer used was 0.1% dinitrochlorobenzene (DNCB) in acetone. The negative control animals were exposed to solvents only. RESULTS General Observations All animals were successfully dosed and appeared normal during the study period. Dermal Observations and Scoring: ‘The results of dermal observations are presented in Table 2. The results of dermal ‘observations from the SRC validation study are presented in Table 3 (A & B), ANALYSIS AND CONCLUSION There was no skin irritation or sensitization observed with either the control or the test groups during the post challenge phase. In the final analysis of data, consideration was given to the overall patterns, intensity, duration, and character of reactions of the test as compared to those of the control animals. In conclusion, the test article (device) did not cause a sensitization reaction under the conditions of this assay.‘SRC Study No. D10971.046.28 Final Report Page 7 of 10 Table 2: Dermal Observations: Challenge Phase (Dermal Scorin; an 24 Hour Score 48 Hour Score ae Right Dose Site Right Dose Site Control Group TFL:352, 0 0 = 1F2:353, 0 0 - 1F3:354 0 0 - TPA355 0 0 = IFS:356 0 0 - 1F6:357 0 0 - ‘Score Incidence (%) 0.0 00 B Sever 0.0 0.0 = Test Group F338 0 0 = 2F2:359 0 0 : 2F3:360 0 0 = 2F4:361 0 0 = 2F5:362 0 0 = 2F6:363 0 0 = SP 1364 0 0 = 3F2:365 0 0 = 3F3:366 0 0 - 3F4:367 0 oO = 3F5:368 0 0 z Score Incidence (%) 0.0 0.0 = Severity Score 0.0 0.0 =‘SRC Study No, D10971,046-28 Final Report 7 Page 8 of 10 ‘Table 3 (A): Positive Control Dermal Observations: Challenge Phase (Dermal Scoring) Animal ID. 24 Hour Score 48 Hour Score Right Dose Site Right Dose Site Results Control Group (Acetone) CIFLI8S CIF2:186 (CIF3:187 CIF4:188 CIFS:189 CIP6:190, ‘Score Incidence (%) 0.0 0.0 Severity Score ‘Test Group (0.1% DNCB) TIFI:I1 TIF2:192 TIF3:193, TIFA194 TIFS:195 TIF6:196 TIFT:197 TIF8:198 TIF9:199 TIF10:200 ee a (as FT TIFIL379 Score Incidence (%) 72.7 81.8 Severity Score 07 08‘SRC Study No. D10971,046-28 Final Report Page 9 of 10 ‘Table 3 (B): Positive Control Dermal Observations: Challenge Phase (Observation Notes) Animal ID Observation Time point C1F5:189 | 3 mm line caused by clippers created after scoring and shaving | 24 hour score Cranial half of dose area patchy erythema 24 hour score TIFI:191 Entire dose site affected 48 hour score Cranial half of dose area patchy erythema 24 hour score TIF2:192 Entire dose site affected 48 hour score TIF3:193. | Small area in cranial-lateral side of dose site affected 48 hour score TIF4:194 | Entire dose site affected 48 hour score 2 patches erythema, center and eranially located 24 hour score TIFS:195 Small area in cranial-lateral side of dose site affected 48 hour score -lateral dose site margin, very discrete erythema 24 hour score TIF6:196 I-lateral side of dose site affected 48 hour score uteral dose site margin, very discrete erythema 24 hour score TLF7:197, Lateral half of dose site affected 48 hour score Patchy erythema across entire dose site 24 hour score TIF8:198 Cranial-lateral side of dose site affected 48 hour score Patchy erythema across entire dose site 24 hour score TIF10:200 Entire dose site affected 48 hour score‘SRC Study No, D10971,046-28 Final Report Page 10 of 10 REFERENCES SRC SOP. ISO 10993-10:2010 ISO 10993-12:2007 FDA 21 CFR-Part 58 USDA Policy 12 Sinclair Research Center, LLC, Standard Operating Procedure Manual Biological Evaluation of Medical Devices Part 10: Tests for Irritation and Skin Sensitization Biological Evaluation of Medical Devices Part 12: Sample Preparation and Reference Materials Good Laboratory Practice For Nonclinical Laboratory Studies Consideration of Alternatives to Painful/Distressful Procedures