Environmental disinfection for coronavirus Revision

10/03/2020
BBZIX® Spain

Resume of microbicidal activity of environmental disinfection carried out by BBZIX disinfectants.

Background:
Novel Coronavirus SARS-CoV-2, the Cause of COVID-19, is a virus causing the 2020 pandemic
outbreak. Disinfection is one of the mayor prevention tools we do have in order to keep the
spreading of the disease under control.
BBZIX is a disinfectant manufacturer company based in Spain. The two disinfectants under this
analysis are Aquazix Plus® and Zix Virox®

Introduction:
Coronaviruses are not as common in human medicine but are more frequently found in animal
production. It is frequently found at poultry production, the Avian Infectious Bronchitis Virus,
which is also a Coronavirus.
Coronaviruses can even cause a certain level of casualties, but they are not very resistant against
the microbicidal activity of many disinfectants.
Microbicidal activity against Novel Coronavirus SARS-CoV-2, has been proved by the collection
of bibliography that shows the activity of the active ingredients at BBZIX disinfectants, against
this Virus and by its comparison against other Coronaviruses, whose virucide activity seems to
be similar.

Active ingredients VIRUCIDE Activity

Hydrogen Peroxide Virucide activity:

SARS: severe acute respiratory syndrome (SARS), was another episode of human coronavirus
that caused an outbreak (2003) disease but with a smaller impact of number of people and
countries affected.
As COVID-19 is very recent, not many studies have been carried out and have been published in
order to be useful for the current outbreak. That’s why the reference for this Virus, would be
SARS outbreak 2003.
Regarding this virus (SARS), some studies 1(A. Schmidt, M.H. Wolff and O. Weber, 2005) have
shown effective disinfection activity of Hydrogen peroxide at a concentration of 0.5%.
This study shows that within less than 3 minutes, 4 Log10 can be reduce of this coronavirus
(SARS) using a concentration of 5,000 ppm or 0.5% of Hydrogen Peroxide.

Furthermore, in another study 2(Anthony F. Henwood, 2020), it is described the microbicide

activity of Hydrogen Peroxide against this virus. The effective virucide activity it is carried out
by a concentration of 0.5% of Hydrogen Peroxide, within 1-minute exposure time.

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We can find more information regarding the Hydrogen Peroxide Virucide activity at 3(Navid
Omidbakhsh, BSc,a and Syed A. Sattar, PhDb, 2005), at this study Hydrogen Peroxide is widely
studied against many pathogens and also against to Human coronavirus 229E-also a surrogate
for the SARS virus (VR-740).
This study shows that a concentration of 0.5% of Hydrogen Peroxide during 1-minute exposure,
is virucide against the Virus. This Virucide activity is quantified as more than 4 Log10 reduction
of infectivity. It was carried out under a presence of 5% of serum (mixture of proteins), in order
to simulate the presence of body fluids but also the level of stringency of methods for added
confidence in the test data.

Peracetic Acid Virucide activity:

Peracetic Acid is a well-known disinfectant, it is known due to its wide spectrum microbicide
activity. A very recent study 4(Minjin Wang et al., 2020.) recommends Peracetic Acid, as an
effective virucide against 2019-nCov Coronavirus,
As it has been said before, coronaviruses are not especially resistant as many other envelop
Viruses. The problem could be found when the virus is under presence of organic material. Some
disinfectants are not effective enough under this condition. Peracetic Acid is highly
recommended 5(Xin-Wei Wang a et al.), for effective disinfection against Coronavirus in
conditions of high levels or organic material as faces or urine.

Cell culture and genome integrity detected by nested RT PCR. Coronavirus SARS-CoV as well as
RSV seem to be sensible to the different disinfectants tested in a study, 6(F. ANSALDI et al. 2004).
This Study was carried out as follows: A known quantity (1ml) of saline solution containing a
standard concentration of cell-grown virus, was mixed with a larger volume (2 ml) of different
disinfectants under evaluation at their use dilution. The mixtures were held for a defined contact
time (30 seconds, 1 minute, 2 min, 5 min, 15 min, 30 min) at controlled temperature (18°C) and
humidity (40%). Peracetic Acid disinfectants were used at a dosage of 0.035%. SARS CoV appears
to be completely inactivated by disinfectants such as Peracetic Acid, after < 1-minute exposition.

Silver Microbicide Activity:

Silver particles have a long history of been well-known for its microbicide activity. Unlike
Hydrogen peroxide and Peracetic acid, studies involving Silver particles, do not talk about
effective dosages against viruses, but they do explain that there is a virucide effect, and its
synergetic action together with other disinfectants, which is the way we will present it as you
will see further in this document, combined with Hydrogen Peroxide.
Having said that, there are many studies, regarding its activity against bacteria, and some against
viruses. The use of Silver is highly recommended as there is not cytotoxicity evidence to humans,
but also can provide with some residual activity on surfaces.

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A study 10(Eric Rentz Do Comm Cnmo (2003)) states that it can be concluded that the smaller
the colloidal Silver particle is, the more biologically active it becomes. So colloidal Silver at
suspension or diluted in a fluid, has more microbicide activity than other bigger particles of
silver.
If indeed the SARS strain of human coronavirus is enveloped, as it is supposed to be, with a
minimal fatty wrapper or without a fatty wrapper, which is more likely, then the oligodynamic
Ag+ should readily denature its envelope, its capsid, its protein constituents, and its entire
genome, as suggested by the work of (Feng et al.)(. Zhang et al.) and as stated at the study
previously mentioned, 10(Eric Rentz Do Comm Cnmo (2003)), largest genome of RNA virions, an
envelope averaging 80–160nm in diameter, thus making it a large target for oligodynamic Ag+
adsorption when presented in picoscalar colloidal form. In the case of SARS, a picoscalar (8A°) 1
oligodynamic Ag+ formula can impregnate up to 30,000 particles into the outer envelope of a
single coronavirus 100nm in diameter.
There is more evidence about how nanoparticle of Silver can bind Virus. One study 11(Stefania
Galdiero 2,3,4 et al., 2011) shows, Silver nanoparticles within the range of 1–10 nm were able
to bind to the virus (HIV), the evidence is clear when it was found that nanoparticles were not
randomly attached to the virus but it was found established regular spatial relationships with
the viral envelope.
This study explains that there is proved virucide activity against HIV-1 [37–40], hepatitis B virus
[41], respiratory syncytial virus [42], herpes simplex virus type 1 [43,44], monkeypox virus [45],
influenza virus [46] and Tacaribe virus [47].
HIV can be related to coronavirus as it also is enveloped and it has capsid.

And a very interesting finding is that Silver together with other disinfectants have shown that
there is an interesting synergetic microbicide activity: This activity has been proven by some
studies for Hydrogen Peroxide (Armon et al. 2000; Rafter et al. 1999). It seems that metal binds
to a biological molecule and is reduced by superoxide radicals or other reductants and then
reoxidized by Hydrogen Peroxide. Continuous redox reactions in a cyclic manner result in
damage, as radical formation occurs near the target site of the molecule (Samuni et al. 1984;
Thurman and Gerba 1989; Yahya et al. 1992).
There are many different mechanisms already proven by evidence as follow:
Affinity for sulfhydryl groups (Davis and Etris 1997¸Feng et al. 2000)
Site-specific Fenton mechanism (Thurman and Gerba, 1989)
Immobilization of the virus to a surface (Thurman and Gerba, 1989)
Blocks or destroys host-cell receptors (Thurman and Gerba, 1989)
Inactivation of the nucleic acid within the viral capsid (Thurman and Gerba, 1989)

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BBZIX DISINFECTANTS COMPOSITION:

Aquazix Plus®:
Hydrogen peroxide 50%. Silver 0.038%. Analysis of peroxides according to: UNE-EN 902,
European Chemical
Codex, Standard of Quality: US and EU Pharmacopoeia. Synergistic combination of stabilized
peroxide. Peroxide manufactured with Food-industry registration.

Zix Virox®:
Hydrogen peroxide 25 %, Peracetic Acid 5 %,

BBZIX DISINFECTANTS MICROBICIDE ACTIVITY:

All our disinfectants are broad spectrum and have been tested for years under UNE Official
effectiveness norms against different types and strains of bacteria, viruses and fungi. Also due
to their action mechanisms, by oxidation, there is no way microorganisms could develop any
kind of resistance to it. Also, it is under the general knowledge that envelop virus, where
Coronaviruses are classified, are less resistant than other pathogens already tested with an
effective Virucide proven efficacy.

Aquazix Plus®:
The main component of Aquazix Plus® is 50% of stabilized Hydrogen Peroxide. The virucide
activity against Coronavirus (mainly SARS as per the bibliography) is proven to be effective at a
dosage of 1%, of Aquazix Plus® in a dilution with water.
As we can see in the previous chapter “Active ingredients VIRUCIDE Activity (Hydrogen Peroxide
Virucide activity)”, is well recorded in some studies, already stated, at this previous chapter but
also at the end of the resume, at the bibliography. In those studies, they all have shown activity
at a concentration of 0.5% of Hydrogen Peroxide. In order to reach this 0.5% of Hydrogen
Peroxide, we need to use a concentration of 1% of Aquazix Plus® as this active ingredient is
concentrated in the product at 50%.
It is also important to consider, that the Virucide activity is greater at this concentration for the
Silver content in the product, 0.038%. The Silver increases the virucide activity, as microbicide
activity to any other pathogen, but there is not specific evident yet, against coronavirus. As it
has been described before, there are many different mechanisms where the Silver can have
virucide efficacy to enveloped viruses, and RNA genetic material. But also, it has been described
that together with Hydrogen Peroxide, it has a proven synergetic effect with a result of a greater
microbicide activity. One of the positive uses for disinfection of surfaces is that silver can provide
with a residual effect on those surfaces for a few hours. Nevertheless, without any Cytotoxicity
for humans.

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Zix Virox®:
As previous composition description. There are two main active ingredients at this product
against Coronavirus.
It is already explained, the Virucide activity against Coronaviruses, by Hydrogen Peroxide and
Peracetic Acid.
Regarding the Peracetic Acid, it was previously stated that at a concentration of 0.035% it is
proven to have effective virucide activity against Coronaviruses. For that reason, in order to keep
this concentration of the active ingredient we need to use a dosage of 0.7%, diluting the pure
product directly with water.
The product also has a 25% of Hydrogen Peroxide, therefore it should be used at a 2% to keep
a 0.5% of pure active ingredient, if it was the case of being the only active ingredient.
But taking in consideration that 0.7% is already effective due to Peracetic Acid composition,
there is no need to use a greater concentration. Furthermore, it would be expected, to have an
effective virucide activity at a concentration under 0.7% against Coronavirus, cause this
concentration is only given by the Peracetic Acid but not together with Hydrogen Peroxide.
Therefore, it should not be recommended a lower concentration for the lack of evidence at
lower concentrations, but at the same time we should consider that the range of efficacy
security at this dosage is much greater.

Additional information:
In order to clarify and to give extra confidence for the use of BBZIX disinfectants, it has recently
been published a list of approved products against the Coronavirus SARS-COV-2 (COVID-19)
7(List N: EPA’s Registered Antimicrobial Products for Use Against Novel Coronavirus SARS-CoV-
2, the Cause of COVID-19, 03/03/2020).
In that list there are several products with similar or different concentrations of the same active
ingredients, that we have been discussed.

Ex. Product a). *Ready To Use

Hydrogen Peroxide …………………………….1.4%
Other Ingredients: ………………………………98.6%
Total: ………………………………………………….100.0%

Ex. Product b).*Ready To Use

ACTIVE INGREDIENTS:
Hydrogen Peroxide……………………………..3.130%
Octanoic Acid………………………………………0.099%
Peroxyacetic Acid ...................................0.050%
OTHER INGREDIENTS: ............................96.721%
TOTAL: ...................................................100.000%

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Ex. Product c).:

Active Ingredients:
Hydrogen Peroxide .................................27.5%
Peroxyacetic Acid ....................................5.8%
Other Ingredients: ...................................66.7%
Total: .......................................................100.0%

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Bibliography:

1- Environmental survival and microbicide inactivation of coronaviruses

Coronaviruses with Special Emphasis on First Insights Concerning SARS 201
ed. by A. Schmidt, M.H. Wolff and O. Weber
© 2005 Birkhäuser Verlag Basel/Switzerland
2- Anthony F. Henwood (2020): Coronavirus disinfection in histopathology,
Journal of Histotechnology
3- Broad-spectrum microbicidal activity, toxicologic assessment, and materials
compatibility of a new generation of accelerated hydrogen peroxide-based
environmental surface disinfectant, Navid Omidbakhsh, BSc,a and Syed A. Sattar, PhDb
Oakville and Ottawa, Ontario, Canada, 2005
4- A precision medicine approach to managing 2019 novel coronavirus pneumonia, Minjin
Wang et al., 2020.
5- Study on the resistance of severe acute respiratory syndrome-associated coronavirus
Xin-Wei Wang a, Jin-Song Li b, Min Jin a, Bei Zhenb, Qing-Xin Kong a, Nong Songa,
Wen-Jun Xiao b, Jing Yin a, Wei Wei b, Gui-Jie Wang b, Bing-yin Si b,
Bao-Zhong Guob, Chao Liu c, Guo-Rong Oua, Min-Nian Wang b,
Tong-Yu Fang d, Fu-Huan Chaoa, Jun-Wen Li a,∗
a Tianjin Institute of Environment and Health, 1 Da Li Road, Tianjin 300050, PR China
b Beijing Institute of Microbiology and Epidemiology, Beijing 100072, PR China
c Beijing Institute of Pharmacology and Toxicology, Beijing 100850, PR China
d BeijingInstitute of Basic Medicine, Beijing100850, PR China
Received 8 December 2004; received in revised form 3 February 2005; accepted 7
February 2005
Available online 20 March 2005
6- SARS-CoV, influenza A and syncitial respiratory virus resistance against common
disinfectants and ultraviolet irradiation
F. ANSALDI, F. BANFI, P. MORELLI, L. VALLE, P. DURANDO, L. STICCHI, S. CONTOS, R.
GASPARINI, P. CROVARI
Department of Health Sciences, University of Genoa
2004
7- List N: EPA’s Registered Antimicrobial Products for Use Against Novel Coronavirus SARS-
CoV-2, the Cause of COVID-19
Date: 03/03/2020
8- Silver Disinfection in Water Distribution Systems. University of Arizona. Silvestry
Rodriguez, Nadia, 2008.
9- No touch’ technologies for environmental decontamination: focus on ultraviolet devices
and hydrogen peroxide systems
David J. Weber a,b, Hajime Kanamori a, and William A. Rutalaa,b. 2016

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10- Eric Rentz Do Comm Cnmo (2003) Viral Pathogens and Severe Acute Respiratory
Syndrome: Oligodynamic Ag+ for Direct Immune Intervention, Journal of Nutritional &
Environmental Medicine, 13:2, 109-118, DOI: 10.1080/13590840310001594061
11- Silver Nanoparticles as Potential Antiviral Agents. Stefania Galdiero 2,3,4, Annarita
Falanga 2, Mariateresa Vitiello 1, Marco Cantisani 2, Veronica Marra 1 and Massimiliano
Galdiero 1,3, 2011.
12- Novel Antimicrobial Surface Coatings And The Potential For Reduced Fomite
Transmission Of SARS And Other Pathogens. Craig Feied, MD © 2004

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