Plasma generated ozone for decontamination
of surfaces
Innovative technology for rapid, safe and chemical-free decontamination using ozone
Declan Diver,
School of Physics & Astronomy, University of Glasgow
& a Founding Director, Anacail
Dekontamineringsdagene 2016 Oslo
Structure of presentation
Topic
slides
Background to our research: plasmas and ozone
6
Applications to food decontamination
12
Applications in medical sector
10
Environmental applications
3
Summary & Acknowledements
3
Total:
34
2
Anacail: origins
 Anacail is a University spin-out from the Astrophysical Plasma
Group, School of Physics & Astronomy, University of Glasgow,
Scotland
 Our technology was a serendipitous find from the development of
a plasma loud speaker…….
Plasma: gas
mixture of neutrals,
ions and free
electrons
- Created by the
partial electrical
breakdown of a
neutral gas (an
extended ‘spark’)
Dekontamineringsdagene 2016 Oslo
Plasma creation
involves localised
heating – a
pressure wave.
Modulate at right
frequencies:
sound!
3
Anacail: concept development
the loud speaker was built as a sparetime curiosity research project (Hugh
Potts)
It was found to produce ozone copiously
and quickly on demand (less desirable
feature of a domestic loudspeaker
system…. despite the flawless frequency
response )
It quickly became apparent that the
device was more useful as an ozone
source than a loudspeaker…
Dekontamineringsdagene 2016 Oslo
The plasma source
is an atmospheric
pressure dielectricbarrier discharge
(DBD) plasma,
designed to
generate ozone
and other radicals
inside a sealed
package, using a
patented electrode
system that
operates from the
package exterior.
The system
operates at 8 kV,
modulated at 1050kHz, & consumes
20-30W power
4
What and Why : Ozone
Why Ozone?
The ideal sterilising agent
Very powerful germicide (better than Chlorine)
Created from oxygen in air – no chemical additives
Unused ozone decays to oxygen – no toxic residuals
In our device it’s created on demand and in situ – no storage or
handling problems….Sustainable and repeatable
Dekontamineringsdagene 2016 Oslo
5
Ozone production: detail
Plasma creates low-energy free electrons e (and
positive ions)
Electron attachment to oxygen molecule causes it
to split: e + O2 -> O + O- (nb this isn’t ionization,
but dissociation)
Negative oxygen ion can combine with oxygen
molecule to produce ozone: O- + O2 -> O3 + e
Ozone is also destroyed by interaction with atoms
and electrons (and indeed, bacterial cells)
Trick is to get the right balance between creation
and destruction of O3
Note our technique is entirely indirect: the
target surface is not exposed to charged
plasma species
Dekontamineringsdagene 2016 Oslo
6
Anacail – Plasma Head with Sealed Package
(1) Before: Sealed package placed under head
(2) Treatment: Head attached, ozone generated inside pack
Ozone head
Ozone head
O2
N2
N2
O2
N2
O2
N2
air
N2
N2
N2
O2
(3) Decontamination: Ozone diffuses through pack
O2
N2
N2
air + ozone
O
2
N2
O3
Dekontamineringsdagene 2016 Oslo
O2
N2
O3
N2
N2
N2
O2
O2
N2
O3
O2
N2
N2
O2
O3
N2
N2
N2
N2
O2
(4) Completed: All unused ozone decays back to oxygen
O2
N2
N2
O2
N2
air
N2
O2
N2
N2
N2
O2
7
Short video showing Anacail operation
please see www.anacail.com for this movie
The device has been designed with high volume manufacture in mind and
uses standard manufacturing techniques.
Dekontamineringsdagene 2016 Oslo
8
Anacail Targets Medical and Food Market Applications
Food
Medical
Food waste reduction
through shelf life extension
treatment for:
• Fruit, Vegetable
• Ready-Meals
• Bakery
• Fish
Point of use high level
decontamination and
sterilisation of small
instruments
• Sterilisation in pack for
storage
• Multiple market segments
Dekontamineringsdagene 2016 Oslo
9
The Food Story
Food Medical
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10
$bn food wastage: extending shelf life is one solution
The Problem: Wasted Food
15m tonnes
£5b
90m tonnes
Target
problematic/high
value markets:
Soft Fruit*
One Key Solution: Shelf Life Extension
Additives
• $300m market
MAP&CAP**
• >$10b market
Target areas
where MAP&CAP
cannot compete
* Table Grapes, blueberries, snacking tomatoes, which will represent 40-50% of European tomato market in 2015
**Modified Atmosphere Packaging & Controlled Atmosphere Packaging
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11
What are the mechanisms for food waste and which of
those is Anacail addressing?
Spoilage Mechanism
Dekontamineringsdagene 2016 Oslo
Anacail Addresses
Mould
√
Yeast
√
Bacteria
√
Goes Dry
x
Goes Soggy
x
12
MAP is dominant packaging competitor but has issues
MAP&CAP
• Dominant Tech
• Introduction or removal of gases
(varies by food type);
infrastructure and gas cost
• Often requires specialised
packaging
• Major players include:
Other Tech
• Food additives: industry wants
to remove or reduce
• Packaging technology eg
“fresh’n smart” Fresha bags
In many cases ozone can work alongside the above
technologies for enhanced shelf life extension
Dekontamineringsdagene 2016 Oslo
13
Bakery Fruit
Anacail can double shelf life
After 20 days
Anacail treated bags are
all mould free
Almost all the untreated
tomato bags have mould
Day 1
Un-treated
Dekontamineringsdagene 2016 Oslo
Treated
Day 17
Un-treated
Treated
All untreated
muffins
mouldy
All Anacail
treated muffins
mould free
http://www.anacail.com/anacail_muffin_demo.mpg
14
Anacail in operation – bakery treatment
please see www.anacail.com for this movie
Dekontamineringsdagene 2016 Oslo
15
Anacail – iterative process for parameters
We have had significant success in extending the shelf-life of packaged
berries and fruit, by inactivating the spoilage organisms
Number of completely mould-free bags
controls
controls
Dekontamineringsdagene 2016 Oslo
controls
controls
16
Anacail – iterative process for parameters
Proportion of mouldy tomatoes (based on individual counts)
Dekontamineringsdagene 2016 Oslo
17
Anacail in operation – Table Grapes
Grapes (green).
Thompson seedless, stored
at room temperature
Dekontamineringsdagene 2016 Oslo
18
Anacail in operation – strawberries, peaches
Strawberries: Pack quality
3.5
Average pack quality
3
2.5
Control
2
1.5
Anacail Treated
1
0.5
0
0
2
4
6
8
10
Peaches: Ambient : Quality
12
days
since
treatment
C
Q
Average quality score
NB ‘quality’ scores are
produced by retailer, not
Anacail
3.5
3
2.5
2
1.5
1
0.5
0
0
5
10
15
20
days since treatment
C
Q
Dekontamineringsdagene 2016 Oslo
19
Anacail – food applications unit
First prototype
Latest in-line machines
Ceramic electrode head
Plasma is the pink-purple glow
note – a thin layer, not a jet!
Dekontamineringsdagene 2016 Oslo
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The Medical Story
Food Medical
“Anacail offers a unique holistic
cleaning, disinfecting and storage
capability ideally suited to
healthcare challenges “
Professor Andrew Smith – Consultant
Microbiologist and Lead Consultant for Medical
Device Decontamination, Institute of Infection,
Inflammation and Immunity, NHS Glasgow
Dekontamineringsdagene 2016 Oslo
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The Market Opportunity
Relevant Global Market Facts
2017 Infection Control Market:
$14b
Sterilisation Equipment Market:
$3b
Low Temperature Sterilisation Devices sold: ~57,000 units/p.a. – CAGR:10%
driven by 4 major players
10%
4%
Advanced Sterilisation Products
Getinge
3M
25%
61%
Steris
However: A $3bn market plus unsatisfied
demand to treat complex instruments
Dekontamineringsdagene 2016 Oslo
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Anacail’s Medical Value Proposition
Known Sterilisation Challenges
Complex, high value instruments
cannot be treated effectively
Ozone is a powerful germicide
safe for eg heat labile surfaces
Centralised treatment centres:
slow turnaround times & duplicate
instruments stock
Distributed Treatment Solution
at point of use - small footprint
Other distributed solutions put
operators at risk eg harsh chemicals
Operator safety by avoiding
aggressive chemicals
Storage difficult: air drying racks
mean once opened, instruments
must be used immediately
Safe storage: in sterile package
until needed (cf endoscope air
drying cabinets)
Difficult to monitor and demonstrate
compliance with regulations
Confirmed decontamination
level data to satisfy regulations
Dekontamineringsdagene 2016 Oslo
23
Prototype medical applications machine
high-level
decontamination
machine for medical
device applications
One such machine is in
Glasgow Dental School
labs for trials
Designed for small-scale
activity, rather than
production line
High level decontamination is > log 6 kill
Dekontamineringsdagene 2016 Oslo
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Effective decontamination of liquid drops
Drops containing
bioluminescent
Salmonella Enteridis
4 to 128 microlitre
drops of LB broth
laced with bacteria
(grown in cultures to
OD600nm of 0.8 )
(model for food and
instrument contamination)
Salmonella Enteritidis was
transformed to express a
bioluminescent phenotype by
electroporation with the plasmid
pGLITE. This plasmid encodes the
luxCDABE operon of Photorhabdus
luminescens under constitutive
control of the lac promoter and
contains a kanamycin (Km)
resistance cassette as the selective
marker
Dekontamineringsdagene 2016 Oslo
Sealed lid
25
Effective decontamination of liquid drops
Inactivated cells collect at centre
Time lapsed light signal from the
largest treated droplet; guide lines
showing the approximate diffusion
speed of ozone into liquid water
-6
cross section (mm)
-4
-2
0
2
4
6
-15
-10
-5
0
5
10
time (min)
15
20
25
30
Dekontamineringsdagene 2016 Oslo Ozone
is effective at decontamination even if present in droplets
26
Initial (un-optimised) trials at Dental School
Treatment of dental forceps
• Three sets put individually into small 50 μm zip-lock
bags, in ambient air.
• Inoculated with Pseudomonas aeruginosa (50ml at 107
/ml) & allowed to dry
• Non-control treated for 100 s (c 4mg ozone); all
sonicated In 10ml PBS
• Liquor syringed out, plated and incubated
Decimal value
log10
avg sample
333
2.52
avg ctrl
4,400,000
6.64
avg log kill
4.12
max sample
min ctrl
min log kill
600
750,000
min sample
max ctrl
max log kill
100
9,900,000
Dekontamineringsdagene 2016 Oslo
2.78
5.88
3.10
Controls at different
dilutions
Treated (undiluted)
2.00
7.00
5.00
27
Current Status and Decontamination Data
Target
Trial 1
log kill
Trial 2
log kill
Test Soil
Inoculate: P
Aeruginosa
6.47
n/a
Endoscope
Inoculate: P
Aeruginosa
5.3
5.24
Contrast between controls and Anacail treated inoculated
samples: test soil and non-lumened endoscopes (Andersen
Caledonia)
Experimental data from various trials (Andersen
Caledonia and Glasgow Dental School
Dekontamineringsdagene 2016 Oslo
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Current Status and Decontamination Data
Surrogate Trial 1
lumens
log
reduction
Trial 2
log reduction
PA
9.51
9.36
CA
9.02
9.0
SA
9.5
>8.92
BS
10.44
>9.6
Dekontamineringsdagene 2016 Oslo
Contrast between controls and plasma
treated lumens, for a variety of pathogen
inoculants. Figures in red denote total kill –
ie no recoverable organisms in the treated
sample (Andersen Caledonia).
29
Medical: D-values in air discharge
Process development
Process developed against Most Resistant Organism (MRO), which presents high
resistance to sterilant Bacillus Stearothermophilus recognised as MRO for ozone
Biological Indicators (BI’s) of this species obtained from different suppliers to check
for consistency.
Bionova and Excelsior are different manufacturers of open BI’s (Stainless disk in
Tyvek envelope)
Treatment was 100s plasma in air, followed by delay shown on graph
D-value is approx. 8-9 mins/log (unoptimised process)
7.0
log 10 remainig spore count
6.0
5.0
4.0
Bionova
3.0
2.0
Excelsior
y = -0.1156x + 5.9726
R² = 0.9653
y = -0.138x + 5.651
R² = 0.9501
1.0
0.0
0
Dekontamineringsdagene 2016 Oslo
10
20
30
40
50
60
time in bag post treatment/min
30
The Anacail Medical Value Proposition
• Time and money saving by avoiding instruments being sent to centralised
sterilization location (nb – washing is still necessary)
• Operator safety by avoiding aggressive chemicals
• Safe storage solution: in a sterile container until needed (different to
endoscope air drying cabinets for example)
• Confirmed decontamination level data to satisfy regulations
• Potential to de-nature prions: Recent studies have shown molecular ozone
could de-nature prions (major implications in dental and neuro surgical
instruments)
• Ozone disables antibiotic traces on surfaces* – useful environmental aspect
• Small footprint for conveniently locating equipment in
hospital/laboratory/other environments
• Compatibility of treatment with instrument surface materials, including
screens
Dekontamineringsdagene 2016 Oslo
*Lange et al Chemosphere 65 17-23
31
Environmental applications
Drain
decontamination
Dekontamineringsdagene 2016 Oslo
 Biofilms in drain wells are a
source of re-infection:
– Listeria in food factories
– Norovirous in Hospitals
(and other buildings)
 Liquid chemicals not effective
in key problem area (drain
well wall)
 Gasesous ozone can control
build up and re-infection
 Anacail has IP filed on
innovative solution (patent
now granted)
 Prototypes being built and
evaluated
32
Decontamination of drains and pipes
log reduction compared to control
7
log reduction: biofilms on
pipe interiors
Biofilm grown on standard plastic
plumbing pipe is destroyed by our
ozone treatment– even under
water!
6
Wet pipe sections
5
4
No recoverable
organisms on dry
sample
3
Biofilms are resistant to liquid
biocides, but are gas-permeable…
Possible areas of application:
drains, taps, shower heads…
Pipe
sample
subjected
to Anacail
ozone
treatment
2
1
0
Dry
Fresh stop end Horizontal rep 1 Horizontal rep 2
Investigation funded by Biomedical Catalyst, InnovateUK + MRC
Dekontamineringsdagene 2016 Oslo
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Summary
Plasmas are used to generate ozone, a gaseous biocide
Generation inside a sealed package, from outside, ensures safety and enduring
decontamination
Plasma is confined to thin layer on container surface – does not impinge on target
Ozone inactivates surface contamination
Ozone decays back to normal oxygen in a short period
Ozone gas penetrates where liquids cannot
Ozone generation uses only the air in the package or sealed container
Plasma generated ozone is a simple and effective surface cleaner
Anacail Team
Executive team
Ian Muirhead (CEO):
20 years in VC backed university spin-out companies
CEO and BD roles in a wide range of technology markets
Hugh Potts (Founder Director & Principal Scientist):
15 years in plasma experimentation and modelling
Since 2009 focussed on commercialising Anacail’s technology
Declan Diver (Founder Director & Scientific Advisor)
Reader in Plasma Physics at Glasgow University, 25 yrs
experience theory & modelling
Non executives & Advisory panel
Gareth Voyle (Chair):
previously CEO Geest, now NED Oscar Mayer
Jonathan Lintott (Sterilisation Advisor)
CEO of Andersen Caledonia, (Medical Device Sterilisation Service company)
Liz Kynoch (Food Advisor)
previously Technical Director, Tesco PLC
Dekontamineringsdagene 2016 Oslo
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Thank you for listening – Takk for lytting!
Acknowledgements
£: Grant funding
SUPA (Scottish Universities Physics Alliance)
STFC IPS (follow-on fund) ,
InnovateUK (Biomedical Catalyst),
TSB (Food manufacturing)
Scottish Enterprise (SMART awards)
££:Our investors:
IP Group, Scottish Investment Bank, Sussex Place Ventures
Plus Andrew Smith (Glasgow Dental School), Meg Pajak & Tony Buckley (P&A, Dental
School), Hugh Potts, Ian Muirhead & Dick Barton (Anacail), Dann Turner & Darren Reynolds
(UWE).
www.anacail.com
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