ANTI-MICROBIAL CONFORM - Graffiti Shield · THE ANTI-MICROBIAL ACTION OF SILVER IONS In its (elemental) metallic form, silver is inert and does not kill bacteria. Silver atoms (written

ANTI-MICROBIAL CONFORMPROTECTIVE FILM

714-575-1100 www.Graffiti-Shield.com

Tested by third party to ISO testing standards against most common illness-causing bacteria and coronavirus. Antimicrobial technology is applied to the product; it does not directly protect the user from any exposure to bacteria or viruses, or provide additional health benefits. Pure Zone antimicrobial film contains an active ingredient (silver ions approved by EPA under file number 73148-1-91742). Pure Zone has been tested and proven to reduce Coronavirus by up to 99.87% (after 1 hour) and reduce up to 99.99% of bacteria (after 1 hour) such as: Listeria, Salmonella, Golden Staph, Methicillin-resistant Staphylococcus aureus, and Pseudomonas aeruginosa. It is strongly recommended to practice good hygiene, wash hands regularly, and maintain social distancing as often as possible.

During cleaning operations, as well as in the presence of humidity, silver ions are released from the top layer of the film. These ions come into contact with the bacteria, blocking their metabolism and/or interrupting their proliferation mechanism, leading to their destruction When the Anti-Microbial film is manufactured, silver ions encapsulated in a glass matrix are distributed over the film in a uniform manner.

Active strains (according to ISO 22196)Reducing > 99.99 % of bacteria ascertained for:

• Escherichia coli• Salmonella (Salmonelle enterica)• Listeria (Listeria monocytogenes)• Golden staph (Staphylococcus aureus)• Methicillin-resistant Staphylococcus aureus (MRSA)• Pseudomonas aeruginosa

Antiviral activity on the Human coronavirus HCoV-229E strain (according to the ISO 21702 standard):

• 94.99 % after a contact time of 15 minutes• 99.87 % after a contact time of 60 minutes

Under normal usage conditions, harmless when in contact with human skin (skin compatibility study carried out under dermatological control). • No allergenic potential • No irritant potential

• Active compound: silver ions, < 0.3 % w/w of the entire product

• Antimicrobial activity maintained after 365 cleanings with water, alcohol, and chlorine bleach

• Resistance to solvents: the adhesive is resistant to most chemicals (alcohol, diluted acids, oils). The film can be cleaned/disinfected by all conventional cleaning methods, using non-abrasive accessories, cleaning products, detergents or products currently used in healthcare environments

• The disinfection levels achieved are compatible with common usage in the most sensitive areas in terms of infectious risks

• Recommended minimum application temperature: +10 °C (+50 °F).

• Operating temperature range: from -40 °C to +90 °C (-40 °F to +194 °F)

ANTI-MICROBIAL CONFORMPROTECTIVE FILM

714-575-1100 www.Graffiti-Shield.com

Anti-Microbial film applied in areas of frequent handling (e.g.: handles) or high visitor frequency will be subject to repeated abrasion which will reduce its durability (change of appearance, lifting, etc.). The film durability depends on the intensity and frequency of the mechanical stress to which it is exposed.

ANTI-MICROBIAL FILM FEATURES:Indicative value

• Thickness: 2.4 mil Average Values

• Tensile strength (N/25 mm): min. 15 • Elongation at break (%): min. 100• Shrinkage 168 hours at 70 °C (158 °F) (mm): < 0.6 mm

ADHESIVE PROPERTIES:(Measured average values at publication of the technical data sheet)

Average value Standard• Peel strength test 180° on glass (N/25 mm): - after 20 minutes of application 15 - after 24 hours of application 17• Initial tack (N/25 mm): 20• Release (N/25 mm): 0.5

THE ANTI-MICROBIAL ACTION OF SILVER IONS

In its (elemental) metallic form, silver is inert and does not kill bacteria. Silver atoms (written as Ag or Ag0) must lose an electron and become positively charged silver ions, Ag+. Silver is ionised in air, but above all in a humid environment. Silver ions are highly reactive and affect multiple sites in bacterial cells, guaranteeing their destruction.

SILVER IONS : • Produce alterations to the

cell wall

• When transported into the cell, bind with proteins and interfere with the production of energy, enzyme function and cell replication

• Silver ions are active on a broad spectrum of bacteria

Reference : CHL-R14-0815 Intertek France - Etablissement de Chalon/Saône CHL-R16-1301 12 rue Alfred Kastler – Boite N° 7 71530 FRAGNES www.intertek-france.com

Intertek France – Etablissement de Chalon/Saône

This report only concerns samples submitted for testing. This report must not be reproduced, if not in entirely, without the written authorization of the laboratory General Conditions of Services information available upon request. You can send your comments on this report in two months by email [email protected]

Certificate of analysis

Society : HEXIS Address : ZI Horizons sud

34110 FRONTIGNAN

To the attention of: Nathalie SIBOLD

Customer Reference: Film Antimicrobien Pure Zone Intertek Sample Reference: 14-CHL-0815-02 / 16-CHL-1301-01Date of Sample Receipt: September 1st, 2014 / November 28th, 2016Date of Sample Analysis: September 10th, 2014 / December 6th, 2016Date of the Certificate of Analysis: July 21st, 2017

Test: Evaluation of antimicrobial efficiency based on ISO 22196

Results: The results are given as log reduction R, corresponding to the value of antimicrobial activity.

Salmonella enterica subsp enterica R > 4.56 (for sample 14-CHL-0815-02) Listeria monocytogenes R > 4.21 (for sample 14-CHL-0815-02) Staphylococcus aureus R > 4.12 (for sample 14-CHL-0815-02) Escherichia coli R = 4.54 (for sample 14-CHL-0815-02) MRSA R > 3.51 (for sample 14-CHL-0815-02) Pseudomonas aeruginosa R > 5.37 (for sample 16-CHL-1301-01)

Certified by Sylvie LEBRAT Laboratory Director

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R2007HEX001-1

Antiviral activity of PUREZONE060/PRZ150/PURECOVER surface on

human coronavirus HCoV-229E for a contact time of 15 and 60min.

Adapted protocol from ISO 21702 (201) standard

CLIENT

HEXIS

Monsieur Christophe BAUDRION

Z. I. Horizons Sud

34110 FRONTIGNAN

FRANCE

PRESTATION

EFFECTUÉE PAR

S.A.S VIRHEALTH

Site Laennec-La Buire, 2éme étage, Bat B

7-11 rue Guillaume Paradin,

69372 Lyon Cedex 08

CONTRIBUTION

TECHNIQUE Léa Szpiro, responsable laboratoire

Dounia Bouchami, technicienne de laboratoire

Approbateur/Validation Qualité

Nom : Dr Vincent Moulès, CEO

Date : At Lyon, 01/07/2020

Signature :

This report includes 10 pages

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SUMMARY

I. CONCLUSION ...................................................................................................................... 3

II. CONTRACTUAL DOCUMENTS ............................................................................................... 4

III. TEST CONDITIONS AND SAMPLES DATA ............................................................................... 4 III.1 SAMPLES IDENTIFICATION ............................................................................................................... 4

II.1 EXPERIMENTAL CONDITIONS ............................................................................................................ 5

III. RÉSULTS ............................................................................................................................. 6 ANTIVIRAL ACTIVITY OF THE PUREZONE060/PRZ150/PURECOVER SURFACE ON HUMAN CORONAVIRUS HCOV-

229E FOR A CONTACT TIME OF 15 AND 60 MINUTES ..................................................................................... 6

a. Cell susceptibility ............................................................................................................................. 6 a. Cytotoxicity ...................................................................................................................................... 6 a. Essai ................................................................................................................................................. 7

VI. ANNEXES ............................................................................................................................ 8 ...................................................................................................................................................................................... 8

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I. CONCLUSION

Antiviral activity of the PUREZONE060/PRZ150/PURECOVER surface and non-active surface have

been tested under conditions defined by the ISO 21702 (2019) adapted protocol for contact times of 15

and 60 minutes on the human coronavirus HCoV-229E

Stainless steel surface is the control for this test.

● PUREZONE060/PRZ150/PURECOVER surface, 15 minutes of contact time

Under experimental conditions, (20°C, 15 minutes), the PUREZONE060/PRZ150/PURECOVER

surface shows an antiviral activity associated with a logarithmic reduction of 1.30 log10 which is

equivalent to a 94.99% efficiency under the ISO 21702 adapted protocol.

● PUREZONE060/PRZ150/PURECOVER surface, 60 minutes of contact time

Under experimental conditions, (20°C, 15 minutes), the PUREZONE060/PRZ150/PURECOVER

surface shows an antiviral activity associated with a logarithmic reduction of 2.90 log10 which is

equivalent to a 99.87% efficiency under the ISO 21702 adapted protocol.

PRODUCT

Contact time (min)

Logarithmic reduction (log10) Antiviral efficiency (%)

PUREZONE060/PRZ1

50/PURECOVER

15

1.3

94,99%

60

2.9

99.87%

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II. CONTRACTUAL DOCUMENTS

The present service is defined by the following contractual documents:

. Quotation DEV0033

. Order Good for agreement date: 03/06/2020

III. TEST CONDITIONS AND SAMPLES DATA

III.1 Samples identification

Test surface: PUREZONE060/PRZ150/PURECOVER (batch N° C014023C)

The test surface (film) is marketed under the following names:

PUREZONE060, PRZ150 et PURECOVER

HEXIS has provided VirHealth with technical data sheets and a certificate attesting that the test

surface evaluated in this test is identical for the 3 references.

Control surface: stainless steel

Product appearance: white, self-adhesive film, smooth and non-porous

Manufacturer: HEXIS

Supplier: HEXIS

Storage conditions: room temperature

Evaluation period: 06/2020

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II.1 Experimental conditions

Test surface: PUREZONE060/PRZ150/PURECOVER

Experimental Conditions

Date - 19/06/2020

Viral strain - Human coronavirus HCoV-229E

Sample size (cm²) - 1.5 cm x 1.5 cm = 2.25 cm²

Inoculum size (cm²) - 1 cm x 1 cm = 1cm²

Inoculum volume - 50uL

Temperature 20°C

Contact time 15 and 60 minutes

Interfering substance n.a.

Neutralisation - 2 mL of infection medium without FCS

Quantification - endpoint titration on permissives cells

Number of wells per dilution 4

Incubation temperature 34 ± 1 °C

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III. RÉSULTS

Antiviral activity of the PUREZONE060/PRZ150/PURECOVER surface on

human coronavirus HCoV-229E for a contact time of 15 and 60 minutes

a. Cell susceptibility

Surface lOG

TCID50/mL

PUREZONE060/PRZ150/PUR

ECOVER 5.1

Stainless steel 5.0

Difference < 1 log10 ☒ yes ☐ no

Comparative titre of human coronavirus HCoV-229E on MRC5 cells inoculated with

PUREZONE060/PRZ150/PURECOVER surface and stainless steel surface recuperation buffer show

a difference less than 1log10. Results showed that recuperation buffers of the test surfaces didn’t affect

the MRC5 susceptibility to human coronavirus HCoV-229E under test conditions.

a. Cytotoxicity

The test surface cytotoxicity is determined by reading of cytopathic effect (CPE) on MRC5 permissive

cells and quantified by TCID50 technique.

For viral recuperation on surface, the surfaces are submerging in 2mL of infection medium without

FCS (recuperation buffer). The recuperation buffer cytotoxicity is determined by reading of cytopathic

effect (CPE)

Under test conditions, the recuperations buffers from PUREZONE060/PRZ150/PURECOVER and

stainless steel surfaces didn’t show cytopathic effects on MRC5 cells for a contact time of 15 and 60

minutes.

The test results are dependant and take into account the cytotoxicity results.

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a. Essai

Raw data for antiviral activity of PUREZONE060/PRZ150/PURECOVER and stainless steel surfaces

on human coronavirus HCoV-229E under test conditions (20°C, 5, 15 and 60 minutes) are presented

in appendices

Results have been determined by visual reading of cytopathic effects (CPE) and quantified by TCID50

technique on MRC5 cells.

Surface Cytotoxicity

(log10 TCID50)

Support T0

(log10 TCID50)

T15 (log10 TCID50)

T60 (log10 TCID50)

PUREZONE060/PRZ150/PUREC

OVER 0.5

S1 5 3,5 2,1

S2 5,1 4 2,4

S3 5,1 4,1 2,4

Average N1 5,1 3,9 2,3

SD 0,1 0,3 0,1

stainless steel 0.5

S1 5 5 5,1

S2 5,3 5,3 5,3

S3 5 5,3 5,1

Average N2 5,1 5,2 5,2

SD 0,1 0,1 0,1

Reduction D1 (log10

DICT50) * / 1,3 2,9

N1: viral quantity in log10 (average of triplicate) stainless steel surface

N2: viral quantity in log 10 (average of triplicate) PUREZONE060/PRZ150/PURECOVER surface

* D: antiviral activity for every contact time (logarithmic reduction in log10) for1 cm2 of test surface

D1=N1-N2

V. CONCLUSION

PUREZONE060/PRZ150/PURECOVER surface shows antiviral activity of 94.99% (1.3

log10 TCID50) and 99.87% (2.90 log10 TCID50) on human coronavirus HCoV-229E after

a contact time of 15 and 60 minutes respectively at 20°C.

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VI. ANNEXES

- Cell line

Name: MRC5 ATCC® CCL-171™

Number of passages: 15

Culture medium: EMEM (Lonza, batch n°0000757679, 11/2020) with 10% of FCS (Dutscher, batch

n° S16529S1810, 09/2022), 1% of antibiotics (Gibco, batch n° 2145466, 12/2020) and 1% of L-

glutamine (Gibco, batch n° 2091579, 22/2021)

- Viral strain

Name: human coronavirus 229E ATCC® VR-740™

Viral test suspension: 7.50x10^7 (batch number: 032020229-4)

Quantification technique:

- Successive tenfold dilution in infection medium: EMEM (Lonza, batch n°0000757679,

11/2020) with 2% of FCS (Dutscher, batch n° S16529S1810, 09/2022), 1% of antibiotics

(Gibco, batch n° 2145466, 12/2020) and 1% of L-glutamine (Gibco, batch n° 2091579,

22/2021)

- Add 100µL of every dilution on 8 wells on a 96 plate.

- Incubate 7 days at 34°C, 5% C02

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● V.3 RAW DATA : TCID50 quantification of human coronavirus 229E

after 15 and 60 minutes, visual reading of cytophatic effetcs (4 wells per

dilutions)

Tableau 1 : stopping activity control

Explanations: - 1-4: degrees of CPE in 8 cell culture unit (microtiter plate)

- 0: no virus present

- n.a: not applicable - n.d: not done

● Tableau 2 : cytotoxcicity

Explanations: - 1-4: degrees of CPE in 8 cell culture unit (microtiter plate)

- 0: no virus present - n.a: not applicable

- n.d: not done

ProductContact

time

P 1 2 3 4 5 6 7

0 44444444 44444444 44444444 44444444 10011010 0 0 0

0 44444444 44444444 44444444 44444444 10110110 0 0 0

0 44444444 44444444 44444444 44444444 10111010 0 0 0

0 44444444 44444444 44444444 44444444 00011110 0 0 0

0 44444444 44444444 44444444 44444444 10111101 0 0 0

0 44444444 44444444 44444444 44444444 01011001 0 0 0

dilutions (-log)

T0

PUREZONE060/PRZ

150/PURECOVER

s ta inless s teel

ProductContact

time

P 1 2 3 4 5 6 7

0 0 0 0 0 0 0 0

0 0 0 0 0 0 0 0

0 0 0 0 0 0 0 0

0 0 0 0 0 0 0 0

0 0 0 0 0 0 0 0

0 0 0 0 0 0 0 0

cytotoxicity

sta inless s teel

/

PUREZONE060/PRZ

150/PURECOVER

dilutions (-log)

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● Tableau 3 : test

Explanations:

- 1-4: degrees of CPE in 8 cell culture unit (microtiter plate)

- 0: no virus present - n.a: not applicable

- n.d: not done

ProductContact

time

P 1 2 3 4 5 6 7

15 44444444 44444444 44444444 0 0 0 0 0

15 44444444 44444444 44444444 10010110 0 0 0 0

15 44444444 44444444 44444444 00011111 0 0 0 0

60 44444444 00101111 0 0 0 0 0 0

60 44444444 11011111 0 0 0 0 0 0

60 44444444 11111110 0 0 0 0 0 0

15 44444444 44444444 44444444 44444444 10011010 0 0 0

15 44444444 44444444 44444444 44444444 10110111 0 0 0

15 44444444 44444444 44444444 44444444 10111110 0 0 0

60 44444444 44444444 44444444 44444444 00011111 0 0 0

60 44444444 44444444 44444444 44444444 10111101 0 0 0

60 44444444 44444444 44444444 44444444 01011001 0 0 0

PUREZONE060/PRZ

150/PURECOVER

dilutions (-log)

sta inless s teel

Essais