TIP - Manila

363 P. Casal, Quiapo

City of Manila, Philippines
Website: www.tip.edu.ph
Tel. Nos. (+632) 733-9117 / (+632) 733-9142

CHE 503: BIOCHEMICAL ENGINEERING

LECTURE 8:
STERILIZATION, DESIGN AND
OPERATION
Lecturer:
ENGR. MICHAEL ALLAN G. RAMOS
Department of Chemical Engineering
Technological Institute of the Philippines
1st Semester, A.Y. 2016-2017

STERILIZATION
Definition
- absence of detectable levels of viable organisms in a culture
medium or gas.
- complete removal of all microorganisms.
- A requirement for an aseptic process.
- reason for sterilization: economics, loss of product due to
contamination is a costly price to pay. No chance of recovery.
Disinfection
- to reduce number of viable pathogenic microorganisms where
they can no longer infect
Pasteurization
- method of reducing the # of viable microorganism from a
product (such as milk or beverage) to extend shelf-life

METHODS OF STERILIZATION
1. Thermal
- preferred for large-scale operation; moist heat usually
- pressurization leading to production of wet steam
- 30psia, 1210C, 15 minutes

2. Chemical
preferred for heat-sensitive equipment
- ethylene oxide (gas) for equipment
- 70% ethanol solution (acidified to pH =2 using HCl) for surfaces of
equipment
- 3% sodium hypochlorite solution for surfaces

3. Radiation UV for surfaces, X-ray for liquids

4. Filtration membrane ultra-filters with 1 nm size to filter off viruses and
bacteria; for oxygen or air sterilization

METHODS OF STERILIZATION
1. sterilization of all equipment especially inside surfaces where
contamination can start at crevices, roughness etc.
2. sterilization of the medium, so that contaminants will not
compete getting food vs. your target or microorganism of choice
3. can be done direct or indirect. Batch or continuous

RESISTANCE OF MOS TO MOIST HEAT

THERMAL METHOD
1. Not all organisms have identical death kinetics.
(increasing difficulty; vegetative cells < spores < virus)
2. Individuals within a population of the same organism respond
differently
Probability Theory:
P(t) = the probability that an individual cell is still viable at time t.

P(t ) = e

k d .t

(simplest form assuming 1st order death kinetics)

- 0 = will not occur; 1 = will occur
kd .t No probability of an
1 Po (t ) = 1 1 e
unsuccessful sterilization

THERMAL METHOD

kd .t No

1 Po (t ) = 1 1 e
Po (t ) = probability of extinction of a population at time=t,
<=1
k d = death rate constant, 1/time (1st order kinetics)
holding time, exposure time, time (temp. dependent)
t=
N o = initial number of viable microorganism at time=0,
# of spores

Use probability chart to evaluate 1-Po(t), 0.001 is the acceptable safe

standard probability

With increments,
0.18,0.018 etc..

Temperature Effects on Kinetics

of Thermal Sterilization
Arrhenius equation

k d = .e
kd =
=
R=
Ed =

Ed
R .T

Death rate constant, 1/time

Frequency factor, 1/time
Gas constant, 8.314 J/kmole-K
Deactivation energy, (50-150 kcal/g-mole = spores)
(2-20 kcal/g-mole = vitamins/ growth factors

T = Sterilization temperature, K

Temperature Effects on Kinetics

of Thermal Sterilization

kd = .e

Ed
R.T

Ed 1
ln k d = ln .
R T

ln k d

THERMAL METHOD
dN
= k d .N
dt
dN
= k d .dt
N
N
ln
= k d .t
No

N = N o .e

k d .t

N = final # of viable MOs

THERMAL INACTIVATION
Goal is to kill microorganisms as contaminants as well as to preserve
the chemical compounds/ nutrients in fermentation broths

HTST
- Degradation of vitamins/ nutrients in the media can be avoided via brief
exposure to high temperature

BATCH vs. CONTINUOUS

HEAT STERILIZATION
BATCH
STERILIZATION

steam out

steam in

BATCH vs. CONTINUOUS

HEAT STERILIZATION
CONTINUOUS
STERILIZATION

Pe/ Da chart

Pe/ Re chart

Use of an equation instead of Re/Pe chart

For

3,500 < N Re < 300,000

Dz
1.3 x10 0.4
+

=
2.195
0
.
04
N

v.D N Re
Re
(Espaldon, 9/25/2012)

Use of an equation instead of Re/Pe chart

4.00

3.50

3.00

2.50
Levenspiel
Dz/v.D 2.00

Aki1
Aki2

1.50

Sjenitzer

1.00

0.50

0.00
0

10000

20000
Nre

30000

REFERENCES
Dr. David Shonnard, Chemical Engineering, Michigan Technological
University
Bioprocess Engineering, Shuler, Kargi
Principles of Biochemistry, Lehninger, 4th edition
Bioprocess Engineering Fundamentals, Pauline Doran
Dr. Loh Kai Chee, Chemical and Biomolecular Engineering, NUS