WO2022248901A1 - 100% natural aqueous-alcoholic gel and solution 100% natural hand sanitizer - Google Patents

Abstract

Virucidal aqueous-alcoholic solution or gel.

Description

Description

Title of the invention: 100% natural hydroalcoholic gel

1. Thanks to Dr Matthieu Rigaud and Sébastien BALDI as well as with the laboratory

SleA, manufacturer of natural sanitary products, following long research we were able to develop the first 100% NAUREL hydro alcoholic gel. Indeed we are the only ones today to manufacture this type of 100% natural disinfection not derived from synthetic products or any additive from petrochemicals. It took us months of research to arrive at a very high efficiency result.

Our Hydro alcoholic gel is not toxic for humans and the environment, is said to be "Eco friendly".

2. Product Texture:

It comes in two very distinct forms, the first has a more viscous appearance like "GEL". It is obtained thanks to a very precise manufacturing process and a gelling agent that is also 100% NATURAL.

The second aspect is in liquid form, the only difference between the two is only pure the aspect of viscosity according to the demand of the users.

The gel from our SleA laboratories will be dedicated for human use only (TPI Classification)) while the Solution can also be used for surfaces (TP2 Classification + TPI Classification)

3. Efficiency test:

Our hydroalcoholic gel and solution has been tested by a private laboratory ANALYTICE, in order to know the effectiveness.Important: the name of our product in the analysis report entitled MEDICLEAN.

The tests have been validated according to European standard EN 14476+ A2:07-2019:

The conclusion report demonstrates that in accordance with EN

14476+A2:07-2019, our Hydroalcoholic Gel and Solution product, is therefore effective on all viruses (including H INI and coronavirus) under the conditions tested: 97%, 1 mm, 20°C and cleanliness (EN 14476+A2 :07-2019).

Our product is VIRUCIDE, FUNGICIDE, BACTERICIDE up to 99.99% from 30 seconds after application.

The full report is attached to the file entitled "R/20/20167V2"

SleA Laboratories

Mr. Sebastien BALDI

42 avenue Irene and Jean Frederic Joliot Curie

SOMMAIRE 83130 THE GUARD

SUMMARY

Page

1. Mission framework 3

2. Identification of samples 3

3. Experimental conditions 3

4. Validation of method 4

4.1. Cytotoxicity 4

4.2. Sensitivity of cells to viruses 4

4.3. Validations of the efficacy of stopping the activity of the product 5

4.4. Reference inactivation test 6

5. Actual tests - Calculation of virucidal activity 7

5.1. For ADENOVIRUS 7

5.2. For POLIOVIRUS 8

5.3. For NOROVIRUS 8

6. Verification of the methodology 9

7. Closing 10

APPENDICES

Appendices: pages 11 to 27 IDENTIFICATION OF SAMPLES

Your order : /

1. Mission framework

SleA Laboratoires asks Analytice to have the virucidal activity of the MEDICLEAN product determined according to standard EN 14476+A2:07-2019: chemical antiseptics and disinfectants - Quantitative suspension tests for the evaluation of virucidal activity in the medical field.

2. Identification of the samples - Manufacturer: SleA Laboratoires, - Date of manufacture: not communicated, - Optimum use-by date: not communicated, - Storage conditions: room temperature and darkness, - Active substances: ethanol, - Appearance: colorless gel. - Thinner recommended by the manufacturer: none, product ready to use. - Date of receipt at the laboratory: 10/07/2020. - Study period: from 07/10/2020 to 08/21/2020.

3. Experimental conditions - Test temperature: 20°C ± 1°C. - Titration method: log TCID50 and log PFU/mL (Poliovirus). - Contact time: 30 s (adenovirus and norovirus) - 1 min (poliovirus). - Final concentration: 97% (modified method), 80% and 10%. - Diluent of the product used during the tests: distilled water. - Viral strains: Adenovirus type 5, adenoid 75 strain, cultured on HEp-2 cells,

Murine norovirus, strain S99, grown in RAW cells

264.7, under 5% CO2 atmosphere, and

Poliovirus type 1 strain LSc-2ab, cultured in cells

VERO. - Interfering substances: cleanliness: 0.3 g/L BSA. - Stability of the product in the presence of interfering substance: good. - Technique for stopping the virucidal action: cold.

Viral title:

Titration by cytopathic effect of the viral test suspensions (calculated according to the Spearman-Kärber method):

- Adenovirus = 6.875 log TCID50

- Poliovirus = 6.680 log PFU/mL

- Murine norovirus = 6,750 log TCID50 4. Validation of the method

4.1. Cytotoxicity

- For the adenovirus, the HEp-2 cells were exposed to the MEDICLEAN product and low toxicity was observed up to the 10 ^-1 dilution.

- For poliovirus, the VERO cells were exposed to the MEDICLEAN product and low toxicity was observed up to the 10 ^-1 dilution.

- For norovirus, the RAW cells were exposed to the MEDICLEAN product and low toxicity was observed up to the 10 ^-1 dilution.

4.2. Sensitivity of cells to viruses

For each of the viral suspensions used during the performance of these tests, comparative titrations of the virus are carried out on the cells treated or not with the product.

The product tested at the concentration indicated above does not show any notable influence on the adenovirus titration method (difference < 1 log).

The product tested at the concentration indicated above does not show any significant influence on the poliovirus titration method (difference < 1 log).

The product tested at the concentration indicated above does not show any significant influence on the norovirus titration method (difference < 1 log). 4.3. Validations of the efficacy of stopping the activity of the product

The method is validated if the difference is < 0.5 log).

FOR ADENOVIRUS

FOR POLIOVIRUS

FOR NOROVIRUS

4.4. Reference inactivation test

FOR ADENOVIRUS

Validation of the tests is effective if the reduction in the viral titer between the control suspension and the suspension subjected to formaldehyde is between 0.5 and 2.5 log after 30 min. The reduction is 1.500 log after 30 min and the conditions of the standard are therefore fulfilled.

FOR POLIOVIRUS

Validation of the tests is effective if the reduction in the viral titer between the control suspension and the suspension subjected to formaldehyde is between 0.5 and 4.5 log after 60 min. The reduction is 3.424 log after 60 min and the conditions of the standard are therefore fulfilled.

FOR NOROVIRUS

Validation of the tests is effective if the reduction in the viral titer between the control suspension and the suspension subjected to formaldehyde is between 0.5 and 2.5 log after 30 min. The reduction is 1.375 log after 30 min and the conditions of the standard are therefore fulfilled.

5. Actual tests - Calculation of virucidal activity

5.1. For ADENOVIRUS

Trial 1

The control viral suspension has a concentration of 6.875 log TCID

The product concentrations tested are virucidal if the reduction in viral titer is greater than or equal to 4.0 log.

Z-test

The control viral suspension has a concentration of 6.875 log TCID ₅₀

The product concentrations tested are virucidal if the reduction in viral titer is greater than or equal to 4.0 log.

5.2. For POLIOVIRUS

Trial 1

The control viral suspension has a concentration of 6.680 log PFU/mL

The product concentrations tested are virucidal if the reduction in viral titer is greater than or equal to 4.0 log.

Trial 2

The control viral suspension has a concentration of 6.680 log PFU/mL

The product concentrations tested are virucidal if the reduction in viral titer is greater than or equal to 4.0 log.

5.3. For NOROVIRUS

Trial 1

The control viral suspension has a concentration of 6.750 log TCID ₅₀

The product concentrations tested are virucidal if the reduction in viral titer is greater than or equal to 4.0 log.

Trial 2

The control viral suspension has a concentration of 6.750 log TCID

The product concentrations tested are virucidal if the reduction in viral titer is greater than or equal to 4.0 log. 6. Verification of the methodology

The tests have been validated according to European standard EN 14476+A2:07-2019:

- The titer of the test viral suspension is high enough to allow a reduction of 4 log to be observed after the tests: º It is 6.875 log TCID50 for the adenovirus. º It is 6.680 log PFU/mL for poliovirus. º It is 6,750 log TCID50 for murine norovirus.

- The difference in titers between viral control and reference virus in the inactivation test is between 0.5 and 2.5 after 30 min (4.5 after 60 min for poliovirus): º The difference is 1,500 log after 30 min formaldehyde inactivation for adenovirus. º The difference is 3.424 log after 60 min of formaldehyde inactivation for poliovirus. º The difference is 1.375 log after 30 min of inactivation by formaldehyde for norovirus.

- The product tested does not significantly affect cell morphology.

- The cleanliness test conditions do not affect the infectivity of the 3 viruses tested.

- The product tested does not reduce the sensitivity of cells to viruses: º For adenovirus, the difference is 0.500 log.

° For poliovirus, the difference is 0.453 log. º For norovirus, the difference is 0.625 log.

7. Closing

The tests carried out on the MEDICLEAN product have demonstrated: . That the MEDICLEAN product, used from 97%, has virucidal activity on poliovirus, adenovirus and murine norovirus, according to the methodology of standard EN 14476+A2:07-2019, for 1 minute of contact at 20°C, in clean conditions.

- That in accordance with standard EN 14476+A2:07-2019, the MEDICLEAN product is therefore effective - That in accordance with standard EN 14476+A2:07-2019, the MEDICLEAN product is therefore effective on all viruses (including H1N1 and coronavirus) under the conditions tested: 97%, 1 min, 20°C and cleanliness (EN 14476+A2:07-2019, point 4, table 1 page 9). . That the MEDICLEAN product, used from 80%, has virucidal activity on murine adenovirus and norovirus, according to the methodology of standard EN 14476+A2:07-2019, for 30 seconds of contact at 20°C, under cleanliness.

- That in accordance with standard EN 14476+A2:07-2019, the MEDICLEAN product is therefore effective on all enveloped viruses (including H1N1 and coronaviruses) adenoviruses, noroviruses and rotaviruses, from 80% for 30 sec of contact at 20°C, under clean conditions (EN 14476+A2:07-2019, point 4, table 1 page 9 "The test relating to "virucidal activity against enveloped viruses [with adenovirus and murine norovirus as model viruses] "will cover all enveloped viruses, adenoviruses, noroviruses and rotaviruses")

APPENDICES

ANNEX 1

Adenovirus: - Cell line: HEp-2 cells (RD-Biotech ref. 84011, batch no. 110315-118) - Viral strain: adenovirus type 5, adenoid strain 75 (ATCC ref. VR-5, batch no. 3679877)

Poliovirus: - Cell line: VERO cells (RD-Biotech ref. 84009, batch no. 110118-110V) - Viral strain: poliovirus type 1, strain LSc-2ab (ref RGB-1260 - batch no. 2/10121998- Friedrich Loeffler Institute)

Norovirus:

- Cell line: RAW 264.7 cells (ATCC TIB-71) - Viral strain: murine norovirus, STRAIN S99 (batch n° 4/200409/220409- Friedrich Loeffler Institut)

Culture buffers and media:

- PBS buffer: sodium chloride, Panreac, ref. 141659.1211, lot no. 0000204679; sodium phosphate dibasic, Sigma Aldrich, ref. S5136, Batch No. BCBC7067V; sodium phosphate monobasic, Sigma Aldrich, ref. S5011, batch no. 1019K01021V

- MEM media, Sigma Aldrich, ref. 0268, lot no. 040M8301 - DMEM media, Sigma Aldrich, ref. D5796, Lot No. RNBB9336

- Fetal Bovine Serum, Sigma Aldrich, F7524, Batch No. 098K3397

Reagents:

- Bovine serum albumin powder, Dominique DUTSCHER, ref. 871001, lot D1304039

Inactivation solution: - Formaldehyde, Sigma Aldrich, ref. F-1635, Lot No. BCBB3510

APPENDIX 2

Table A1 - Titration of adenovirus by cytopathic effect, by Spaerman-Kärber calculation method:

Log DICT ₅₀ = 6.875

Figure 1 - graphical representations of test results:

Trial 1

Tableau A2 - Tableau de résultats du produit MEDICLEAN soumis à essai contre l’adénovirus en conditions de propreté (0,3 g/L BSA)

Trial 2

Table A2 - Table of results of the MEDICLEAN product tested against adenovirus in clean conditions (0.3 g/L BSA)

Table A3 — Raw data of MEDICLEAN product tested against adenovirus, under clean conditions (cytopathic effect titration; 8 wells)

ESSAI 2

ESSA1 1

TEST 2

Table A4 - Titrage du norovirus murin par effet cytopathique, par la méthode de calcul Spaerman- Karber : Sensitivity of cells to the virus:

Table A4 - Titration of murine norovirus by cytopathic effect, by Spaerman-Karber calculation method:

Log DICT ₅₀ = 6,750

Figure 2 - graphical representations of test results:

Trial 1

Tableau A5 — Tableau de résultats du produit MEDICLEAN soumis à essai contre le norovirus murin en conditions de propreté (0,3 g/L BSA)

Trial 2

Table A5 — Table of results of the MEDICLEAN product tested against murine norovirus under clean conditions (0.3 g/L BSA)

Table A6 — Raw data of MEDICLEAN product tested against murine norovirus, under clean conditions (cytopathic effect titration; 8 wells)

ESSAI 2

ESSA1 1

TEST 2

Table A7 - Titrage du poliovirus (essai sur plaque) :

Susceptibility of cells to the virus:

Table A7 - Poliovirus titration (plate assay):

Los PFU/mL= 6.680

Figure 3 - graphical representations of test results: Test 1

Trial 2

Table A8 — Table of results of the MEDICLEAN product tested against poliovirus under clean conditions (0.3 g/L BSA)

Table A9 — Raw data of MEDICLEAN product tested against poliovirus under clean conditions.

ESSAI 2

Sensibilité des cellules au potiovirus :

TEST 1

TEST 2

Sensitivity of cells to potiovirus:

4. The positive effects of our formula:

• Not aggressive for the skin

• Non-toxic for the environment

• Ingredients from agriculture (beetroot alcohol, cereal alcohol, etc.)

• Natural product

• Usable in pregnant women (no contraindication)

• Can be used in children (from 2 years old)

• Destroys 99.9% of bacteria

• Does not weaken the hydrolipidic film

Composition of our product:

1. List of our ingredients:

Alcohol: The alcohol we use comes from agriculture, our process is 100% vegetable and 100% natural,

Distilled Water: The distilled water we use is deionized water with a conductivity of less than 1.25 μS/cm. It is accredited with the ISO 9001 certificate.

100% Vegetable Glycerin: A moisturizing agent whose effectiveness on cells has been scientifically demonstrated, vegetable glycerin improves the appearance of the skin, making it supple and smooth. It is non-comedogenic and very low allergenic.

Hydrosols: All our hydrosols are 100% pure, without preservatives. They are microfiltered at 0.2 microns for optimal cleanliness and certified microbiologically compliant. They come mostly from plants from certified organic farming and ecological cultivation.

Product classification:

Our products are classified in TP1 and TP2

To know:

TP1: Includes human hygiene products that are not cosmetics due to their disinfection purpose.

TP2: Includes disinfectants and algaecide products not intended for direct application on humans or animals.

We are the only ones to manufacture a TP1, TP2 product without synthetic ingredients.

Permission

All our products are declared and validated by the authorities, including the national health agency (ANSES). no. DI-21-01096 and no. DI-21-01097.

Claims

CLAIM: Hydro alcoholic gel and Hydro alcoholic solution 100% natural. This 100% natural gel does not currently exist, we are the first to have found the stable formula.