Water Activity & pH

Measurement for Food Safety

Retail Food Systems Research Conference

IFT Foodservice Division
January 7, 2008

Anthony J Fontana Jr. Ph.D.

Senior Research Scientist
Decagon Devices, Inc.

Decagon Devices, Inc.

 Outline
ƒ Introduction
ƒ pH
ƒ What is it?
ƒ pH Measurement
ƒ Water activity
ƒ What is it?
ƒ Water activity vs. Moisture content
ƒ Water activity Measurement
ƒ Water activity & pH for safety
ƒ Conclusion

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 Introduction
ƒ Water
ƒ Unique molecule - abundant in all 3 phases
ƒ Essential for Life
ƒ Water affects taste, structure, &
susceptibility to spoilage
ƒ Fresh foods (meat, fruit, veg.) contain large
amount of water
ƒ Need to control water in food for safety and
storage
ƒ Control water through pH
and/or water activity

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 Introduction
δ+H Hδ+

O 2δ- H2O + H2O <-> H3O+ + OH-

ƒ Water
ƒ able to Hydrogen bond 3-dimensionally
with other water molecules
ƒ types of bonding with other molecules:
ionic bonds, hydrogen bonds, dipole-
dipole, van der Waals

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 Introduction

ƒ Food Safety
ƒ Control Pathogenic Bacteria:
ƒ pH
ƒ Water Activity
ƒ Temperature
ƒ Potentially Hazardous Food
(Time/Temperature Control for Safety Food).
ƒ Except a FOOD that because of the interaction of
its aW and pH values is designated as not
supporting pathogenic microorganism growth or
toxin formation.

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 Outline

ƒ Introduction
ƒ pH
ƒ What is it?
ƒ pH Measurement
ƒ Water activity
ƒ What is it?
ƒ Water activity vs. Moisture content
ƒ Water activity Measurement
ƒ Water activity & pH for safety
ƒ Conclusion

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 pH

ƒ 1901 – Fritz Haber

ƒ German chemist
ƒ Voltage at certain glass surfaces changed with
the acidity of a solution
ƒ 1909 – Soren Sorensen
ƒ Danish biochemist
ƒ Developed the pH scale
ƒ Defined pH as minus logarithm base 10 of [H+]
ƒ ‘p’ – German word for ‘power’

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 2005 Food Code Definition

ƒ "pH" means the symbol for the negative

logarithm of the hydrogen ion
concentration, which is a measure of the
degree of acidity or alkalinity of a
solution.
ƒ Values between 0 and 7 indicate acidity
and values between 7 and 14 indicate
alkalinity. The value for pure distilled
water is 7, which is considered neutral.

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 pH Definition
ƒ pH scale from 0 to 14

ƒ pH 0 – means a very high acid activity

ƒ [H3O+] > [OH-] acidic
ƒ Lemon juice & vinegar (pH 2 to 3), Stomach acid (pH 1),
Nitric acid & Hydrochloric acid (pH 0)

ƒ pH 7 – neutral
ƒ [H3O+] = [OH-] neutral
ƒ Water

ƒ pH 14 – very low active acid concentration

ƒ [H3O+] < [OH-] basic
ƒ Sea water (pH 8), household ammonia (pH 11), oven
cleaners (pH 13), sodium hydroxide (pH 14)

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 pH Definition

ƒ pH = - log [H+]

More accurate:

ƒ pH = - log aH+

ƒ aH+ = hydrogen ion activity

ƒ Activity = effective concentration of H+
ƒ Other ions shield H+ and affect their
ability to participate in chemical reactions

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 pH Measurement
ƒ Indicator Dye
ƒ Certain organic dye solutions change color over a
small pH range
ƒ Indicate approximate pH of a solution
ƒ Dyes are deposited on a strip of paper
ƒ pH Meter / Electrode System
ƒ In an unknown solution the voltage on the outer
electrode surface changes proportionally to changes
in [H+]
ƒ Compares the potential of unknown [H+] to a known
reference potential
ƒ pH meters convert the voltage ratio between a
reference and sensing half-cells to pH values

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 pH Measurement

ƒ Nernst equation
E = Eo + (2.3RT)/nF log{unknown [H+]/internal [H+]}
ƒ E = total potential difference (in mV)
ƒ Eo = reference potential
ƒ R = gas constant
ƒ T = Temperature in Kelvin
ƒ n = number of electrons
ƒ F = Faraday’s constant
ƒ [H+] = hydrogen ion concentration

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 pH Electrodes
ƒ Glass Electrodes
ƒ Combination electrode with both half cells in
one body in either glass or epoxy body
ƒ General purpose glass – pH 0-14, temp to 100°C
ƒ Blue glass – pH 0-13, temp to 110°C
ƒ Amber glass – pH 0-14, temp to 110°C, low Na+ error
ƒ Types: Solution vs gel filled
ƒ Ag/AgCl
ƒ Most common - Suitable for almost all applications
ƒ Temp limit 80°C
ƒ Calomel – Hg/Hg2Cl2
ƒ For samples containing proteins, organics, or heavy
metals that could react with silver and clog the
reference junction
ƒ Temp limit 70°C
ƒ Storage – in pH storage solution (never dry)
ƒ Not water as will cause ions to leach from glass bulb
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 pH Electrodes

ƒ ISFET Electrodes
ƒ Ion-specific field effect transistor
ƒ Non-glass measuring surface
ƒ Semiconductor electrodes on microchip
ƒ Store dry

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 pH Measurement

ƒ pH is a function of temperature
ƒ Voltage output from the electrode
changes linearly in relation to pH and
the temperature of the solution
determines the slope of the graph
ƒ 25°C = 59.16 mV / pH unit
ƒ 0°C = 54.20 mV / pH unit
ƒ 100°C = 74.04 mV / pH unit

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 pH Measurement
ƒ Rinse electrode with dH2O and blot dry
ƒ Note: Never wipe the electrode to remove excess
water – wiping can create static charges that interfere
with pH measurement
ƒ Place electrode in standard buffer – adjust pH
meter to pH value of standard buffer
ƒ Rinse electrode with dH2O and blot dry
ƒ Place electrode in test solution
ƒ Record pH of the test solution
ƒ Remove electrode from test solution and rinse
with water

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 Accurate pH Measurement
ƒ Components
ƒ Electrode
ƒ Calibration (one-point vs two-point)
ƒ 90% problems are related to the improper use, storage
or selection of electrodes
ƒ Cleaning
ƒ Meter
ƒ Temperature compensation (either automatic or
manual)
ƒ Buffers
ƒ Use fresh buffer solutions
ƒ Buffers should frame the range of pH for the samples
being tested
ƒ Buffers should be at the same temperature as the
samples

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 Outline

ƒ Introduction
ƒ pH
ƒ What is it?
ƒ pH Measurement
ƒ Water activity
ƒ What is it?
ƒ Water activity vs. Moisture content
ƒ Water activity Measurement
ƒ Water activity & pH for safety
ƒ Conclusion

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 2005 Food Code Definition

ƒ "aw" means water activity which is

a measure of the free moisture in a
FOOD, is the quotient of the water
vapor pressure of the substance
divided by the vapor pressure of
pure water at the same
temperature, and is indicated by
the symbol AW.

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 Water Activity Definition

Water Activity is a measure of the energy

status of the water in a system.

Old definition: Water activity is the amount of

“free” or “available” water in a product as
opposed to “bound” water.

“Free”: same physical properties as pure water

“Bound”: different physical properties

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 Water Binding Properties

ƒ Factors that Control Water Activity

ƒ Colligative effects or Solute interactions
ƒ Matrix effects or Surface interactions
ƒ Capillary effects

It is a combination of these factors in an

ingredient or product that reduces the
energy of the water as compared to pure
water.

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 Water Activity Definition

ƒ How does water get “Bound” in a

product?
ƒ Ionic bonds
ƒ Hydroxyl groups of sugars
ƒ Carbonyl and amino groups of proteins
ƒ Other polar sites that hydrogen bond
ƒ Dipole-dipole forces
ƒ Van der Waals forces

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 Water Activity Definition

Vapor pressure of water above sample @ °C

aw = Vapor
———————————————
pressure of pure water @ same °C

aw = p/po = %ERH/100

Range: 0 (bone dry) — 1.0 (pure water)

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 Water activity vs. Moisture Content

ƒ Moisture Content
ƒ Quantitative amount of water in a sample on
a wet or dry basis.
ƒ An extensive property that depends on the
amount of material.
ƒ Water Activity
ƒ A measure of the energy status of the water
in a system (Qualitative).
ƒ A intensive property that does not depend
on the amount of material.

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 Cheese and Cracker System

ƒ Experiment 1 - Cracker is equilibrated

over saturated NaCl in a sealed container

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 Cheese and Cracker System

Initial Final
Moisture Content 4% 20%
Water Activity 0.30 0.75

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 Cheese and Cracker System

ƒ Experiment 2 - Cheese is equilibrated

over saturated NaCl in a sealed container

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 Cheese and Cracker System

Initial Final
Moisture Content 60% 30%
Water Activity 0.90 0.75

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 Cheese and Cracker System

ƒ Experiment 3 - Cracker and Cheese are

placed together in a sealed container

20% Moisture 30% Moisture

Which way does water move?

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 Cheese and Cracker System

Equilibrium Conditions
aw cracker = aw cheese = aw air
At equilibrium the energy status
(chemical potentials) are the same
and thus no net exchange of water.
µcracker = µcheese = µair

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 Water activity vs. Moisture content

Traditional Fruit Cake

make components to same
water activity (flour, sugar,
candied fruits, raisins,
butter, eggs, nuts)

Water activity Moisture Content

(% db)
Dough 0.857 24.5
Fruits (mixed) 0.862 52.2

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 Water activity vs. Moisture content

ƒ Differences in water activity between

components, layers, or individual
ingredients leads to moisture migration
and potential problems within a product.

ƒ At equilibrium the energy status of the

water is the same and thus, no net
exchange of water.

aw component 1 = aw component 2

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 Water Activity vs. Moisture Content

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 Measurement of Water Activity

ƒ Water Activity is measured by

equilibrating a product with the vapor
phase and measuring the relative humidity
of the vapor phase.

water vapor
liquid
(humidity)

Sample

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 Water Activity Measurement

ƒ Water activity measurement methods

ƒ Hair or Polymer Hygrometers
ƒ Freezing Point Depression
ƒ Isopiestic Equilibration
ƒ Electric Hygrometers
ƒ Chilled Mirror Dew Point

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 Electrical Properties Sensor
ƒ Moisture changes the electrical
properties of a hygroscopic material
in equilibrium with air above sample.
ƒ Two types:
ƒ Resistance
ƒ Capacitance

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 Electrical Properties Sensor

ƒ Advantages
ƒ Accuracy = ±0.01aw
ƒ Relatively insensitive to volatiles
ƒ Measures entire aw range

ƒ Disadvantages
ƒ Needs calibration (secondary method)
ƒ Need temperature control or
compensation of sensor
ƒ Some sensor hysteresis

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 Chilled Mirror Dewpoint
ƒ Mirror is chilled until dew is formed. The
temperature at which saturation is
achieved is determined by observing
condensation on a chilled surface
(mirror).
Fan
Optical Sensor

Mirror
Infrared Sensor

Sample

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 Chilled Mirror Dewpoint

ƒ Advantages
ƒ Primary method of measuring vapor
pressure (not calibrated)
ƒ Highest accuracy ±0.003aw
ƒ Rapid measurement <5 minutes
ƒ Measures entire aw range (0.03 – 1.0aw)
ƒ High reliability

ƒ Disadvantages
ƒ Need clean mirror
ƒ Readings affected by ethanol and propylene
glycol (>1%)

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 Accurate Measurements

ƒ Calibration / Verification Standards

ƒ Saturated Salt Slurries
ƒ Unsaturated Salt Solutions
ƒ Sample Preparation
ƒ Need representative sample
ƒ If slicing / grinding – be consistent
ƒ Prevent moisture exchange with
environment
ƒ Temperature

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 Outline

ƒ Introduction
ƒ pH
ƒ What is it?
ƒ pH Measurement
ƒ Water activity
ƒ What is it?
ƒ Water activity vs. Moisture content
ƒ Water activity Measurement
ƒ Water activity & pH for safety
ƒ Conclusion

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 Microbial Growth

ƒ Scott (1953) showed that

microorganisms have a limiting
water activity level below which
they will not grow.

ƒ Water activity, not water content,

determines the lower limit of
available water for microbial
growth.

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 Microbial Growth
ƒ Growth Limit
ƒ Every microorganism has a water activity level below
which it cannot grow.

ƒ aw Limit
ƒ 0.94 Growth and toxin production for all
types of Clostridium botulinum
ƒ 0.90 Growth of pathogenic bacteria
except for S. aureus (aerobic)
ƒ 0.88 “Practical” limit for yeast
ƒ 0.86 Staphylococcus aureus (aerobic)
ƒ 0.80 Production of mycotoxins
ƒ 0.70 “Practical” limit for the growth of mold
ƒ 0.60 Absolute limit for the growth of any
microorganism

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 Microbial Growth
ƒ Minimum aw Limit of Growth for Bacteria
Microorganism Minimum aw
Clostridium botulinum E 0.97
Pseudomonas fluorescens 0.97
Escherichia coli 0.95
Clostridium perfringens 0.95
Clostridium botulinum A, B 0.94
Salmonella spp. 0.95
Vibrio parahaemoliticus 0.94
Bacillus cereus 0.93
Listeria monocytogenes 0.92
Bacillus subtilis 0.91
Staphylococcus aureus (anaerobic) 0.90
Staphylococcus aureus (aerobic) 0.86

* Optimum conditions for pH, temperature, etc.

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 Water activity & pH for Safety

ƒ In the United States alone annually due to

foodborne disease :
ƒ 76 million illnesses
ƒ 325,000 hospitalizations
ƒ 5,000 deaths
Source: Centers for Disease Control (CDC)

ƒ The annual medical costs, productivity

losses, and costs of premature deaths due to
five major foodborne pathogens are
estimated to be $ 6.9 billion.
(Crutchfield and Roberts, 2000 FoodReview 23(3):44-49)

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 Microbial Growth

ƒ Hurdle technology
ƒ combines existing and new preservation techniques to
establish a series of preservative factors (hurdles)
that the microorganisms in question are unable to
overcome (jump over).
ƒ These hurdles may be:
ƒ water activity
ƒ pH / acidity
ƒ temperature
ƒ redox potential
ƒ preservatives
ƒ others
Leistner, L., (1994) Further developments in the utilization of hurdle technology for food preservation.,
Journal of Food Engineering, 22:421-432.

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 Microbial Growth

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 Interactive Table A
Table A. Interaction of pH and aw for control of spores in food
heat-treated to destroy vegetative cells and subsequently
packaged.
aw Values pH Values
4.6 or less > 4.6 – 5.6 > 5.6
0.92 or less Non-PHF* Non-PHF Non-PHF
/non-TCS** /non-TCS /non-TCS

> 0.92 – 0.95 Non-PHF Non-PHF PA***

/non-TCS /non-TCS
> 0.95 Non-PHF PA PA
/non-TCS
* PHF means “Potentially Hazardous Food”
** TCS means “Time/Temperature Control for Safety Food”
*** PA means “Product Assessment Required”

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 Interactive Table B
Table B. Interaction of pH and aw for control of vegetative cells
and spores in food not heat-treated or heat-treated but not
packaged.
aw Values pH Values
< 4.2 4.2 – 4.6 > 4.6 – 5.0 > 5.0
< 0.88 Non-PHF* Non-PHF Non-PHD Non-PHF
/non-TCS** /non-TCS /non-TCS /non-TCS
0.88 – 0.90 Non-PHF Non-PHF Non-PHF PA***
/non-TCS /non-TCS /non-TCS
>0.90–0.92 Non-PHF Non-PHF PA PA
/non-TCS /non-TCS
> 0.92 Non-PHF PA PA PA
/non-TCS
* PHF means “Potentially Hazardous Food”
** TCS means “Time/Temperature Control for Safety Food”
*** PA means “Product Assessment Required”

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 Conclusion
ƒ Water Activity and pH are critical for
food safety.
ƒ Both aw & pH are easily and accurately
measured with instrumentation
ƒ aw = p/po
ƒ pH = -log aH+

Decagon Devices, Inc.

 Thank you

Anthony J. Fontana Jr. Ph.D.

anthony@decagon.com

Decagon Devices, Inc.

2365 NE Hopkins Court
Pullman, Washington 99163
Phone: (509) 332-2756 / (800) 755-2751
Fax: (509) 332-5158
Email: aqualab@decagon.com
Web: www.decagon.com/aqualab/

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