Thermal Death Kinetics

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KINETICS OF

THERMAL
REDUCTION
PHRM 433
OUTLINE

 Decimal reduction time (D-value)


 Z-Value
 Survival factor
 Inactivation factor
 First-order thermal specific death rate
 Arrhenius equation
 Worked examples
DECIMAL REDUCTION TIME (D-VALUE)

106 D is the time required for one log reduction (90% kill)
Can be calculated using:
Δt
DT=
log N1-logN2
10 5
# Bacteria

100oC Δt: total exposure time


N1: initial population
1 log N2: population size after treatment
104 T= applied Temperature
D100

103

Time
Example 1:

 calculate the D value for a bacterial suspension of 109 cfu/ml


that was subjected to 85˚C for 15 minutes at which point its density
was reduced to 106 cfu/ml.

Δt Δt: 15 minutes
DT= N1: 109 cfu/ml
log N1-logN2
N2: 106 cfu/ml
15 T= 85˚ C
D85=
log 109-log106

15
D85=
9- 6

D85= 5 minutes
Example 2:

 the D90 value for a bacterium is 2 minutes. If starting culture has


108 cfu/ml, how long should this suspension be kept at 90C
to kill the entire population?

Δt Δt: ? minutes
DT= N1: 108 cfu/ml
log N1-logN2
N2: 1 cfu/ml
Δt T= 90˚ C
2=
log 108-log101

Δt
2=
8- 0

Δt = 16 minutes
The D value: an index for sensitivity to thermal killing

• Which one is more sensitive to heat killing at 100˚C?


Bacillus subtilis or E.coli?
106
• At 1000C the time required to reduce Bacillus population
is longer than that required for E.coli

105
# Bacteria

104
DE.coli DB.subtilis

100oC
103

Time
The D value is temperature dependent

D value decreases as the temperature increases


106 ie. there is less time required to reduce
the population by one log at higher temperatures

105
# Bacteria

D120 D110 D100


104

120oC 110oC 100oC


103

Time
Kinetics of thermal reduction: the Z value

Z value
100 increase in temperature required to reduce
D by 1/10 (one log reduction)
D value (min)

ΔT
Z=
log D1-logD2
10
ΔT: Temperature change
1 log D1: initial D value
D2: secondary D value
1
Z =10˚C

100 105 110 115 120


Temperature (T)
Kinetics of thermal reduction: the Z value

 by having D values for different temperatures

 one can seek for altering the sterilization protocol to fit


to the industrial setting

 One question would be: how much the temperature can


increase to reduce the D value to a given length

 This would provide a pragmatic approach in setting up


SOPs in industrial settings
The use of Z value
 Example:
 A food processing company produces canned meat. Prevention of
Clostridium botulinum spores from growing is important. The D 121 for
botulinum spores is 0.2 minutes and the Z value is 10˚C. the company
wants to sterilize the canned food at 111˚C. what should be the length of
sterilization if they consider to kill 1012 spores per can content.

since every 10˚C decrease in treatment causes 10-fold increase in D value


then:
D111= D121x10 ie. D111= 0.2x10 = 2 minutes
Δt Δt
using, D111= log10 -log10
12 0
2=
12- 0

Δt= 2x12= 24 minutes


They should heat treat their product at 111˚C for 24 minutes.
Problem : try this on your own

The Z value for a microorganism is 2oC. it takes 54 minutes at 75oC to reduce the
population from 109 to 106. At what temperature should the microorganism be
treated to achieve the same result in 10.8 sec.

Answer=800C
 The thermal death kinetics may be represented by the following equation:
-dN/dt = kd N 1
 Where,
 N, is the number of viable organisms present,
 T, is the time of the sterilization treatment
 kd, is the reaction rate constant of the reaction,
or the specific death rate per time.
 On integration of equation (i) from t=0’ to t=t,we have the following expression :

Nt/N0 =e-kdt 2

 where
 No is the number of viable organisms present at the start of the sterilization treatment,
 Nt is the number of viable organisms present after a treatment period, t.
On taking natural logarithms, equation (2) is reduced to:

ln(Nt/N0) = - kd t 3
The graphically equations (1) and (3) are represented as,

Fig: Plots of the proportion of


survivors and the natural logarithm of
the proportion of survivors in a
population of microorganisms
subjected to a lethal temperature over
a time period.

The relationship observed in the above graph would be found only with the
sterilization of a pure culture in one physiological form, under ideal sterilization
conditions.
From Principles of Fermentation
Technology,- Peter F. Stanbury,
Allen Whitaker, Stephen J. Hall,
Second Edition,
Thermal Death Kinetics:
14
dnt
= - k d nt (1)

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dt

Jan
where
nt is the number of live organisms present
t is the sterilization time
kd is the first-order thermal specific death rate

kd depends on the type of species, the physiological form of the cells, as well as the
temperature.

kd for vegetative cells > kd for spores > kd for virus


(10 to 1010/min) (0.5 to 5/min)
Thermal Death Kinetics :
Integrating (1) using the initial condition n = no at t = 0 gives 15
t

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n (2)
ln = - kd dt
t

no

Jan
0
t

n
no
t
= exp - kd dt
0
( ) (3)

Survival factor

1 no
Inactivation factor ≡ = nt
Survival factor
Thermal Death Kinetics (isothermal operation):
kd is a function of temperature, and therefore it is a constant for isothermal operations. 16

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(2) therefore gives

Jan
n (4)
ln = - kd t
t

no

n
= exp(- kd t)
t
(5)
no
Thermal Death Kinetics
(non-isothermal operation): 17
kd is expressed by the Arrhenius equation given below:

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R. Shanthini
( )

Jan
Ed
kd = kdo exp - RT
(6)

where
kdo Arrhenius constant for thermal cell death
Ed is the activation energy for thermal cell death
R is the universal gas constant
T is the absolute temperature
Thermal Death Kinetics
(non-isothermal operation): 18
When kd of (6) is substituted in (2), we get the following:

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R. Shanthini
Jan
t

 ( )
Ed
n - (7)
ln -k
t
exp dt
no = do RT
0

To carry out the above integration, we need to know how the temperature (T) changes with
time (t).
Determining the Arrhenius constants:
19
kd = kdo exp - ( ) Ed

RT
(6)

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Jan
Ed
ln(kd) = ln(kdo) - RT
(7)

ln(kd) ln(kdo)

Ed
R

1/T
Example 1:
A fermentation medium contains an initial spores concentration of 8.5 x 10 10. The medium is
20
sterilized thermally at 120oC, and the spore density was noted with the progress of time as
given below:

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Time 0 5 10 15 20 30

Jan
(min)
Spore density 8.5 x 4.23 x 6.2 x 1.8 x 4.5 x 32.5
(m-3) 1010 109 107 106 104
a) Find the thermal specific death rate.
b) Calculate the survival factor at 40 min.
Solution to Example 1:
Data provided: no = 8.5 x 1010
21
nt versus t data are given

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R. Shanthini
Isothermal operation at 120oC.

Jan
a) Since it is an isothermal operation, thermal specific death rate (kd) is a constant.
Therefore, (4) can be used as follows:

nt
ln = - kd t + c
no

Plotting ln(nt /no) versus t and finding the slope will give the numerical value of kd.
Solution to Example 1:
t (min) 22
0 10 20 30
0

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Jan
-5
ln(nt/no)

-10
-15
y = -0.7201x
-20 2
R = 0.9988
-25

kd = -slope = 0.720 per min


Solution to Example 1:
23
1

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0.8

Jan
0.6
nt/no

0.4
0.2
0
0 10 t (min) 20 30
Solution to Example 1:
b) Since kd is known from part (a), the survival factor at 40 min can be calculated using
24
(5) as follows:

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nt
= exp (- 0.720 per min x t)

Jan
no

= exp (- 0.720 per min x 40 min)

= 3.11 x 10-13 = survival factor

nt = 3.11 x 10-13 no = 3.11 x 10-13 x 8.5 x 1010 = 0.026

We know that nt cannot be less than 1.


The above number is interpreted as the chances of
survival for the living organism is 26 in 1000.
Example 2:
The thermal death kinetic data of Bacillus stearothermophilus (which is one of the most heat- 25
resistant microbial type) are as follows at three different temperatures:

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Temperature (oC) 115 120 125

Jan
kd (min-1) 0.035 0.112 0.347
a) Calculate the activation energy (Ed) and Arrhenius constant (kdo) of the thermal
specific death rate kd.
b) Find kd at 130oC.
Solution to Example 2:
26
Data provided: kd versus temperature data are given

a) Activation energy (Ed) and Arrhenius constant (kdo) of the thermal specific death rate

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(kd) can be determined starting from (10.7) as follows:

Jan
Ed
ln(kd) = ln(kdo) -
RT

Plot ln(kd) versus 1/T (taking T in K).

Slope gives (–Ed/R) and intercept gives ln(kdo).


Solution to Example 2:
27
1/T (per K)
0.0025 0.00252 0.00254 0.00256 0.00258 0.0026

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0

Jan
-0.5 y = -35425x + 87.949
-1 R2 = 1
-1.5
ln(kd)

-2
-2.5
-3
-3.5
-4
Slope = –Ed/R = –35425 K
Intercept = ln(kdo) = 87.949
Solution to Example 2:
28
Slope = –Ed/R = –35425 K

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Ed = (35425 K) (R) = 35425 x 8.314 kJ/kmol = 294.5 kJ/mol

Jan
Intercept = ln(kdo) = 87.949 Activation energy

kdo = exp(87.949) = 1.5695 x 1038 per min


= 2.616 x 1036 per s

Arrhenius constant
Solution to Example 2:
29
b) Since activation energy (Ed) and Arrhenius constant (kdo) of the thermal specific
death rate (kd) are known from part (a), kd at 130oC can be determined using (6) as

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follows:

Jan
kd = kdo exp
( ) -
Ed

RT

= (2.616 x 1036 per s) exp


( -
294.5 x 103
8.314 (273+130) )
= 0.0176 per s = 1.0542 per min
Solution to Example 2:
30
1/T (per K)
0.00245 0.0025 0.00255 0.0026

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0.5

Jan
0
y = -35484x + 88.101
-0.5
-1 R2 = 1
ln(kd)

-1.5
-2
-2.5
-3
-3.5
-4

Calculated value at 130oC

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