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REPUBLIC of TURKEY
ISTANBUL UNIVERSITY
ISTANBUL FACULTY of MEDICINE
Department of Virology and Basic Immunology
Number: 307
Subject: Anti-viral activity report Istanbul, 24th September 2009
To whom it may concern:
The report, related to the studies performed in our laboratory regarding the anti-viral activity of Am-500 may kindly be found in the attachments.
For your kind attention.
Yours sincerely,
Prof. Dr. Selim BADUR
Istanbul Faculty of Medicine
Department of Virology and Basic Immunology
Academic Member

RESULTS OF the TEST PERFORMED in ACCORDANCE with RULES OF “Virus Killing Quantitative Suspension Experiment for Chemical Disinfectants and Antiseptics Used in Medicine” belonging to TURKISH STANDARDS INSTITUTION (TSE) with the number TS EN 14476 (March 2007)
Tested product: Am-500
Virus used for the test: Influenza A (H1N1) Brisbane Strain
Date of report: 24th September 2009
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A. TEST LABORATORY
Istanbul University,
Istanbul Faculty of Medicine, Department of Virology and Basic Immunology
Influenza Reference Laboratory
Capa, 34390, Istanbul
Phone: (212) 635 2882
B. SAMPLE DEFINITION
1. Name of tested product: Am-500
2. Lot number: 06083524
3. Date of expiry: 06/2010
4. Producing company: Indusco Distribution Ltd.
5. Date of production: 06/08
6. Storage conditions: Room temperature
7. Form of product: Liquid
8. Company which sent the product: Nano MSK Technology Marketing Co. Ltd.
C. TEST ENVIRONMENT
1. Test request date: 25.08.2009
2. Temperature of test environment: Room temperature (20oC)
3. Virus titration method: Dilution
4. Concentration at which the disinfectant tested: 20%
5. Contact time of disinfectant and influenza A/H1N1: 1 minute, 10 minutes and 60 minutes
6. Special environmental conditions: PBS (clean environment without any organic debris), 0.3% BSA, 0.3% washed sheep erythrocytes (in order to create a dirty environment with organic debris)
7. Test environment: MDCK cell culture
8. Dilution buffer: Sterile distilled water
D. SUMMARY of METHOD
MDCK cells: Confluent MDCK cells were used for production of virus and to perform the experiment.
Detection of Virus Titration: Influenza A/H1N1 virus was inoculated after serial dilutions and the virus titer was detected based on the virus dilution which caused a clear cytopathic effect that can be visible in an invert microscope.
Detection of Sub-Cytotoxic Concentration of Disinfectant: The concentration of disinfectant recommended by the producer was serially diluted and the concentration which has no toxic effect in cell culture was determined and used in the experiment.

Controls: As negative controls, confluent MDCK cells having neither viral inoculation nor addition of disinfectant as well as virus titration control and controls containing also toxic levels of disinfectant were used.
Clean Environment: Environment containing no organic debris.
Dirty Environment: Environment containing organic debris.

E. TEST VALIDATION
1. Virus titer: log 7.8 TCID50/100μl influenza A (H1N1) Brisbane strain
2. Toxicity experiment: Effects of 10 times serially diluted disinfectant in MDCK cell culture
3. Formaldehyde: 1.4% formaldehyde

F. TEST RESULTS
Virus titer before application: 7.8
Effect of Am-500 (20%)
1 minute
10 minutes
60 minutes
Virus titer after application*
≤2.8
≤2.1
≤1.8
Decrease in virus titer* (log)
≥5.0
≥5.7
≥6.0
*Logarithmic TCID value of virus in 100μl

G. DECISION
The efficacy of 20% Am-500 as a SURFACE PROTECTOR against influenza A (H1N1) has been investigated in accordance with the TS EN 14476 (March 2007) standards of Turkish Standards Institution (TSE) and it has been found that the aforementioned product has decreased the virus titer 5 log, 5.7 log and 6 log respectively in a clean environment when applied for 1, 10, 60 minutes at room temperature (20oC). According to the international standards, a disinfectant must decrease the virus titer 3 log in order to conclude that it is an efficient disinfectant. The results of this experiment indicate that Am-500 is an EFFICIENT SURFACE PROTECTOR against Influenza A/H1N1.

H. SIGNATURE
Meral Akcay Ciblak, DVM, MS Prof. Dr. Selim Badur
Chief of Laboratory Department of Virology and Basic Immunology Academic Member