Sterilization on a practical basis means the absence of any detectable viable organism. : only the...
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Transcript of Sterilization on a practical basis means the absence of any detectable viable organism. : only the...
Sterilization on a practical basis means the absence of any detectable
viable organism.
: only the desired organism is detectably present.
Sterilization methods:
Medium & equipment: .
Heat-sensitive materials in medium: (microfiltrator pore sizes<0.2μm).
Heat-sensitive equipment: UV radiation (energy intensive) or chemical agents (toxic to the desired culture. e.g.70% ethanol-water, and ethylene oxide (gas).
Gas (air): sterilized through adiabatic compression process for supplying oxygen (~2200C) followed by (e.g. glass wool filter).
SterilizationThermal deactivation
The probability of an unsuccessful sterilization:
1- P0(t)
The probability of extinction (successful sterilization):
P0(t)=[1-p(t)]N0
where p(t) is the probability that an individual will still be viable at time t. N0 is the number of individuals (cells or spores) initially present.
Assume first order death rate:p(t)=e-kdt
1- P0(t)=1-[1-p(t)]N0 = 1-[1-e-kdt]N0
where the specific death rate kd = αe-E0d/RT(1/time), constant at specified T. Eod is the activation energy for the death of the organism.
Sterilization
(D): time for the number of viable cells to decrease tenfold.
E[N(t)] = N0 p(t)
E[N(t)] = N0 e-kdt
0.1= e-kdD
D=2.303/kd
E[N(t)] is the expected value of the number of individuals present at time t.
Sterilization1- P0(t)= 1-[1-e-kdt]N0
kd = αe-E0d/RT
From the above equation:- Known N0, T, t, determine Kd, the probability of
an unsuccessful sterilization is determined.
- Given N0, T, acceptable probability of failure e.g. 10-3, required time can be determined
- Higher Kd tends to achieve low probability of sterilization failure. Normally at 121oC.
Kd of vegetative cells > 1010 min-1, spores 0.5-5 min-1. The major concern is spores.
Sterilization Charte.g. N0=108,Kd=1min-1 (1210C), t=26minKdt=26Probability of failure:1-P=0.001
e.g. N0=108,Kd=1min-1 (1210C), 1-P=0.001 Kdt=26Required sterilization time:t=kdt/kd=26 min
(M. Shular, Textbook, p.319)
Batch Sterilization
Temperature verse time in a batch sterilization process
T (K)
t time
heat
ing
holding
cooling
Simplified calculations for deactivation of spores and medium components
T (K)
t time
Kd1
Kd2
Kd3
SterilizationDegradation of components in the medium in
sterilization process.
vitamin and growth factor
The inactivation of viability is much more sensitive to temperature changes than the degradation of important growth factors in the medium
It is important to assure complete killing of foreign organisms without the destruction of important components in the medium.
SterilizationDegradation of components in medium
Assume the degradation rate of such components is first order.
dC/dt=-kdC
The degradation rate constant kd can also be related to temperature by Arrhenius equation.
Integrating the above equation, ln C/C0=-kdt or C=C0e-kdt
where C0 is the initial concentration of the component.
To determine the components remaining active:
the temperature T → determine kd → with known t, determine C.
T (K)
t time
Kd,1
Kd,2
Kd,3
SterilizationDegradation of components in the medium.
33,
23tkdeCC
C0 C1
C2C1
C2 C3
kd = αe-E0d/RT
Δt1
Δt2
Δt3
12,
12tkdeCC 11,
01tkdeCC