Virus Inactivation Test Report

KIWITA
Jan 15
3 min read

No.: 14028212001-01

Date: April 28, 2014

Test Report

Client:

APA Corporation

Test Substance:

Water-Soluble Silicon UMO Concentrated Solution

Title:

Virus Inactivation Test

The sample listed above, submitted to our center on March 18, 2014, was tested.

When publishing this report elsewhere, please follow the publication rules of the testing center.

1. Client

APA Corporation

2. Test Substance

Water-Soluble Silicon UMO Concentrated Solution

3. Test Objective

To evaluate the inactivation effect of the test substance on feline calicivirus.

4. Test Overview

The test substance was mixed with a suspension of feline calicivirus to prepare the reaction solution. The mixture was allowed to act at room temperature, and viral infectivity was measured at 5, 15, and 60 minutes after mixing. A preliminary test was conducted in advance to confirm the measurement method for viral infectivity.

Note: Feline calicivirus is widely used as a surrogate virus for norovirus, which cannot be cultured in cells.

5. Test Results

1) Preliminary TestIt was confirmed that diluting the reaction solution 100-fold with maintenance medium allowed viral infectivity to be measured without interference from the test substance.

2) Measurement of Viral InfectivityThe results are shown in Table 1.

Table 1 – Viral Infectivity of Reaction Solution

TCID₅₀: Median Tissue Culture Infectious Dose, 50% tissue culture infectious dose
Start: TCID₅₀ of the control measured immediately after mixing
Control: Purified water
Virus Suspension: 10× diluted with purified water
Reaction Temperature: Room temperature
<2.5: Not detected
–: Not performed

1 Logarithmic value of TCID₅₀ per 1 mL of reaction solution2 Surrogate virus for norovirus

6. Test Method

1) Test VirusFeline calicivirus F-9 ATCC VR-782

2) Host CellsCRFK cells (Crandell-Rees Feline Kidney) [Dainippon Pharmaceutical Co., Ltd.]

3) Culture Media

Cell Growth Medium: Eagle MEM medium (Nissui) with 10% fetal bovine serum
Maintenance Medium: Eagle MEM medium (Nissui) with 2% fetal bovine serum

4) Preparation of Virus Suspensioni) Cell Culture: Cells were grown as a monolayer in tissue culture flasks using the growth medium.

ii) Virus Inoculation: After removing growth medium, virus was added to the flask, followed by maintenance medium, and incubated at 37°C ± 1°C with 5% CO₂ for 1–5 days.

iii) Preparation of Virus Suspension: After observing cytopathic effects (CPE) under an inverted phase-contrast microscope, the culture medium was centrifuged (3000 rpm, 10 minutes), and the supernatant was diluted 10-fold with purified water to prepare the virus suspension.

5) Test Operation1 mL of test substance was mixed with 0.1 mL of virus suspension to prepare the reaction solution. After incubation at room temperature for 5, 15, and 60 minutes, the mixture was diluted 100-fold with maintenance medium, and viral infectivity was measured. Purified water was used as a control, with measurements at start and 60 minutes.

6) Measurement of Viral InfectivityCells were grown as a monolayer in 96-well microplates using growth medium. After removing the growth medium, 0.1 mL of maintenance medium was added per well. The reaction solution and control, after 10-fold serial dilution, were inoculated into 4 wells each. Plates were incubated at 37°C ± 1°C with 5% CO₂ for 4–7 days. Cytopathic effects were observed under an inverted phase-contrast microscope, and TCID₅₀ was calculated using the Reed–Muench method. Viral infectivity was then expressed per 1 mL of reaction solution.

Note: This paper is translated from the following URL. The content is provided for reference on the scientific research of the raw material only. Whether APA raw materials are used or not, we hope this research will help increase understanding and awareness of body minerals.