Dimas Firmanda Al Riza, ST, M.Sc
TPPB 9
INSTRUMENTASI BIOPROSES
CTD.
Dimas Firmanda Al Riza,
ST, M.Sc
INSTRUMENTASI BIOPROSES
PROSES BIOLOGIS
PEMODELAN
MATEMATIKA
PENGUKURAN SAAT
PROSES
BERLANGSUNG
Sulit memprediksi
dengan akurat
meski cukup
bermanfaat
KOMPLEKS
MULTIVARIABEL
VARIASI
PROPERTIES
INOKULAN TINGGI
KINETIKA REAKSI
TIDAK DAPAT/SULIT
DITENTUKAN
KARAKTERISASI
REAKSI BIOKIMIA
KOMPETITIF
(Vogel and Todaro, 1997)
PENGUKURAN ADALAH
KUNCI UNTUK MEMAHAMI
BIOPROSES!
Teknologi Pengukuran dapat diklasifkasikan menjadi tiga kategori
besar (dapat pula merupakan klasifkasi variabel proses fermentasi):
1. Biological: laju pertumbuhan, forescence, dan laju sintesis
protein;
2. Chemical: konsentrasi glukosa, DO – oksigen terlarut, pH,
konsentrasi gas keluaran CO2, O2, N2, ethanol, ammonia dan
bermacam konsentrasi substansi organik;
3. Physical: Temperatur, level, tekanan, fow rate dan massa.
METODE-METODE
PENGUKURAN PARAMETER
BIOPROSES
1. Biosensor
7. Pengukuran kemurnian air
2. Pengukuran massa/kepadatan
sel/jumlah bakteri
8. Pengukuran Temperatur
3. Pengukuran komposisi bahan
kimia
10.Pengukuran massa
4. Pengukuran DO
5. Pengukuran/analisis exhaust
gas
6. Pengukuran pH
9. Pengukuran tekanan
11.Pengukuran laju massa dan
volume (fow rate)
12.Pengukuran broth level
BEBERAPA HAL YANG HARUS
DIPERHATIKAN DALAM PENGUKURAN
PARAMETER PROSES BIOLOGI
1. Pengkondisian Lingkungan yang STERIL mencegah kontaminasi
2. Alat pengukuran INLINE harus memenuhi “AAA Sanitary Standard”, untuk
spesifkasi permukaan luar dan material pada bagian “basah” (dalam)
3. Material harus tahan terhadap sterilisasi dengan uap panas (steam
sterilization) yang diperlukan secara periodik untuk mencegah penumpukan
bakteri
4. Alat yang terletak pada line proses harus sesuai dengan koneksi sanitari
untuk memfasilitasi pemindahan selama pembersihan ekstensif dan operasi
steril.
5. Port untuk pengambilan sampel, yang digunakan untuk pengambilan bagian
kecil dari isi reaktor untuk analisis laboratorium harus dilengkapi dengan
sistem sterilisasi untuk mencegah kontaminasi mikroorganisme dari luar
selama pengambilan sampel
CONTOH SENSOR UNTUK
PENGUKURAN PARAMETER
FERMENTASI
Culture
parameters
Sensor
Range
Accuracy
Temperature
Pt-100
0-150C
0.1C
Pressure
Piezoresistor
0-2 bar
20 mbar
Gas fow
Thermal
mass fow
meter
Upto 2 v v-1 min
Based on
bioreactor
volume
0.1%
pH
pH electrode
2-12
0.02
pO2
Polarografc
electrode
0-400 mbar
2 mbar
pCO2
Membrane
covered pH
electrode
0-100 mbar
2 mbar
PELETAKAN SENSOR PADA
BIOREAKTOR
Application
measurement
Lower ports
Upper ports
Headplate ports
Temp./pH/DO/ DCO2/
Turbidity
probes/substrate (s)
on-line
+ (for pilot
scale
bioreactors)
-
+ (only for small scale
bioreactors / Glass
bioreactors)
Pressure gauge or
Pressure sensor
-
-
+
Line to outgas
analysis
-
-
+ (after condenser and
out-gas fleter)
Feeding / acid-based /
Antifoam addition
-
Highly suitable (Small
Highly suitable (Large
scale pilot)
scale)
Less Suitable (Large scale) Less Suitable (Small scale
pilot)
Harvest
(Continuous Culture)
(Repeated batch)
+
-
-
BIOSENSOR
BIOSENSOR
Biosensors adalah istilah untuk mengkombinasikan substrat biologis pada
rangkaian elektronik. Hal ini merupakan generasi baru untuk pengukuran
analitis interaksi biomolekular spesifk.
"...a self-contained integrated device which is capable of providing specifc
quantitative or semi-quantitative analytical information using biological
recognition element which is in direct spatial contact with a transducer
element." – IUPAC (International Union of Pure and Applied Chemistry)
Immobilize one of
interacting molecul
(Ligand) onto inert
substrate
Substrat is bounded
onto metal surface
(gold or platinum)
Convert to electricity
signal
METODE TEKNIK
TRANSDUSER BIOSENSOR
1. Potentiometric or amperometric, where a chemical
or biological
reaction
produces a potential
difference or current fow across a pair of
electrodes.
2. Enzyme thermistors, where the thermal effect
of the chemical or biological
reaction is
transduced into an electrical resistance change.
3. Optoelectronic, where a chemical or biological
reaction evokes a change in light transmission.
4. Electrochemically
sensitive
transistors
whose
PRINSIP KERJA BIOSENSOR
PROBLEM 1
Sebutkan contoh jenis BIOSENSOR
dan penerapannya untuk sistem
instrumentasi bioproses!
MASSA SEL
CELL MASS MEASUREMENT
Pengukuran massa sel antara lain dapat dilakukan dengan metode
optical density. Pengukuran jenis ini paling efektif untuk bakteri
yang memiliki bentuk spheris misal E. coli.
Metode lain yang digunakan diantaranya adalah titrasi
spektrofotometri
Optical density (OD) =
A/L
Where A refers to
absorbance
ONLINE CELL MASS
DETERMINATION METHOD
Principle
Advantages
Disadvantages
Optical density
Wide linear range
Some interference
Culture fuorescence
Measurement of cellular
activity
Singanal interpretation
is difcult
Capacitance
Wide measurement range
Interferance from
aeration and aeration
Signal interpretation is
difcult
Measurement of cellular
activity
Ultrasonic
Wide linear range
Self cleaning
Interference from
aeration and agitation
Temperature sensitive
ONLINE CELL MASS AND
VIABILITY MEASUREMENT
Theory:
Cells with intact plasma membranes
act like tiny capacitors under the
infuence of electric feld. The nonconducting
nature
of
plasma
membranes allows charge to build
up. The resulting capacitance can
be measured and is usually
expressed
in
pico-farad
per
centimeter (pF/cm). It depends on
the cell type and is directly
proportional to membrane bound
volume of viable cells.
KOMPOSISI KIMIA
PENGUKURAN KOMPOSISI
KIMIA
Metode yang paling banyak digunakan adalah
kromatograf
Gas chromatography
Liquid Chromatography
Low Pressure
High Pressure
Thin Layer
DISSOLVED OXYGEN
DO
DO merupakan salah satu parameter terpenting yang perlu diamati
dalam proses fermentasi. Parameter ini mengindikasikan potensi
kembang biak bakteri.
Dua prinsip operasi digunakan pada probe pengukuran DO yaitu:
reaksi elektrokimia dan prinsip amperometrik (polarographic).
DO-METER
PRINSIP KERJA PROBE DO
In the amperometric (polarographic) approach, oxygen again
permeates a diffusion barrier and encounters an electrochemical
cell immersed in basic aqueous solution. A potential difference of
approximately 1.3 V is maintained between the anode and cathode.
As the oxygen encounters the cathode, an electrochemical reaction
occurs:
The hydroxyl ion then travels to the anode where it completes the
electrochemical reaction process:
The concentration of oxygen is directly proportional to the amount
of current passed through the cell.
HUBUNGAN ANTARA DO DAN
TEMPERATUR
EXHAUST GAS
ANALYSIS
ANALISIS GAS EXHAUST
Banyak yang bisa dipelajari dari gas
hasil metabolisme seperti CO2, O2, N2,
NH3 dan ethanol. Faktanya analisis
prediktif didasarkan pada penghitungan
Laju uptake oksigen, laju pertukaran
karbon dioksida yang menggambarkan
proses respirasi.
Metode yang digunakan secara luas
adalah spektrometri massa. Dua metode
operasi digunakan yaitu:
Magnetic
Quadrapole
MAGNETIC
QUADRAPOLE
Menggunakan elektron beam
untuk ionisasi partikel.
The quadrapole is a set of four
similar and parallel rods (see Fig.
4) with opposite rods electrically
connected. A radio frequency and
dc charge of equal potential, but
opposite charge, is applied to each
set of the rods. By varying the
absolute potential applied to the
rods, it is possible to eliminate all
ions except those of a specifc
mass-to-energy ratio.
PENGUKURAN PH
NILAI PH UNTUK BEBERAPA
CAIRAN
PH DAN KONSENTRASI ION
HIDROGEN
PENGUKURAN PH
EFEK TEMPERATUR PADA
PENGUKURAN PH
TABEL ERROR TEMPERATUR
JURNAL –
PENGUKURAN PH FERMENTASI – NDT
ULTRASONIK
BROTH LEVEL
As the broth in a fermenter or bioreactor becomes more
viscous and is subjected to agitation from sparging (the
introduction of tiny sterilized air bubbles at the bottom of the
liquid) and from mixing by the impeller, it has a tendency to
foam. This can be a serious problem as the level may rise to
the point where it enters the exhaust gas lines clogging the
ultraflters and possibly jeopardizing the sterile environment
within the reactor. Various antifoam strategies can be
employed to correct this situation, however, detection of
the condition is frst required.
SENSOR ANTIFOAM
STANDAR
Type of foam:
Early foaming
Late foaming
Antifoam efects on:
DO
Growth morphology
(Filamentous MO)
Foam sensors:
Low foam sensor
High foam sensor
PROBLEM 2
Jelaskan mengapa strategi antifoam
diperlukan, dan jelaskan cara kerja
sensor untuk pengukurannya!
INSTRUMENTASI UNTUK
PENGUKURAN PROSES
FERMENTASI AKTIF
CONTOH DATA HASIL
PENGUKURAN
PENGGUNAAN DATA
PENGUKURAN BIOPROSES
Data hasil monitoring bioproses antara lain digunakan untuk:
1. FAULT ANALYSIS
2. PROCESS MODELING
3. STATE ESTIMATION
4. CONTROL SYSTEM
FAULT ANALYSIS
FAULT ANALYSIS
Faults in reactor operation affect 15-20% of fermentations [25].
Fermentation measurements can be used to detect faults; for example,
signals from a fow sensor could be used to detect blockages in a
pipe and trigger an alarm in the factory control room. Normally
however, the sensors themselves are the most likely components to
fail; rates of failure of some fermentation instruments are listed in
Table 13.2.
PEMODELAN
BIOPROSES
PEMODELAN BIOPROSES
Model secara umum adalah hubungan matematika dari variabel2
proses.
As an example, a frequently-used mathematical model for batch
fermentation consists of the Monod equation for growth and an
expression for rate of substrate consumption as a function of
biomass concentration:
STATE ESTIMATION
PENDUGAAN KONDISI
PROSES
Terkadang ada beberapa variabel yang tidak
dapat diukur secara langsung pada bioproses
maka diperlukan estimasi.
Often, considerable delays are involved in
off-line measurement of important variables
such as biomass, substrate and product
concentrations.
Such delays make effective
control of the reactor difcult if control action
is dependent on the value of these parameters,
but must be undertaken more quickly than
off-line analysis allows. One approach to this
problem
is to
use
available
on-line
measurements
in
conjunction
with
mathematical models of the process to
estimate unknown variables.
Program komputer yang dikembangkan untuk ini
dinamakan software sensor, estimator atau
observer.
SISTEM KONTROL
UMPAN BALIK
PEMODELAN BIOPROSES
Jenis sistem kontrol yang digunakan umumnya adalah feedback
control system.
INSTRUMENTASI BIOPROSES
CTD.
Dimas Firmanda Al Riza,
ST, M.Sc
INSTRUMENTASI BIOPROSES
PROSES BIOLOGIS
PEMODELAN
MATEMATIKA
PENGUKURAN SAAT
PROSES
BERLANGSUNG
Sulit memprediksi
dengan akurat
meski cukup
bermanfaat
KOMPLEKS
MULTIVARIABEL
VARIASI
PROPERTIES
INOKULAN TINGGI
KINETIKA REAKSI
TIDAK DAPAT/SULIT
DITENTUKAN
KARAKTERISASI
REAKSI BIOKIMIA
KOMPETITIF
(Vogel and Todaro, 1997)
PENGUKURAN ADALAH
KUNCI UNTUK MEMAHAMI
BIOPROSES!
Teknologi Pengukuran dapat diklasifkasikan menjadi tiga kategori
besar (dapat pula merupakan klasifkasi variabel proses fermentasi):
1. Biological: laju pertumbuhan, forescence, dan laju sintesis
protein;
2. Chemical: konsentrasi glukosa, DO – oksigen terlarut, pH,
konsentrasi gas keluaran CO2, O2, N2, ethanol, ammonia dan
bermacam konsentrasi substansi organik;
3. Physical: Temperatur, level, tekanan, fow rate dan massa.
METODE-METODE
PENGUKURAN PARAMETER
BIOPROSES
1. Biosensor
7. Pengukuran kemurnian air
2. Pengukuran massa/kepadatan
sel/jumlah bakteri
8. Pengukuran Temperatur
3. Pengukuran komposisi bahan
kimia
10.Pengukuran massa
4. Pengukuran DO
5. Pengukuran/analisis exhaust
gas
6. Pengukuran pH
9. Pengukuran tekanan
11.Pengukuran laju massa dan
volume (fow rate)
12.Pengukuran broth level
BEBERAPA HAL YANG HARUS
DIPERHATIKAN DALAM PENGUKURAN
PARAMETER PROSES BIOLOGI
1. Pengkondisian Lingkungan yang STERIL mencegah kontaminasi
2. Alat pengukuran INLINE harus memenuhi “AAA Sanitary Standard”, untuk
spesifkasi permukaan luar dan material pada bagian “basah” (dalam)
3. Material harus tahan terhadap sterilisasi dengan uap panas (steam
sterilization) yang diperlukan secara periodik untuk mencegah penumpukan
bakteri
4. Alat yang terletak pada line proses harus sesuai dengan koneksi sanitari
untuk memfasilitasi pemindahan selama pembersihan ekstensif dan operasi
steril.
5. Port untuk pengambilan sampel, yang digunakan untuk pengambilan bagian
kecil dari isi reaktor untuk analisis laboratorium harus dilengkapi dengan
sistem sterilisasi untuk mencegah kontaminasi mikroorganisme dari luar
selama pengambilan sampel
CONTOH SENSOR UNTUK
PENGUKURAN PARAMETER
FERMENTASI
Culture
parameters
Sensor
Range
Accuracy
Temperature
Pt-100
0-150C
0.1C
Pressure
Piezoresistor
0-2 bar
20 mbar
Gas fow
Thermal
mass fow
meter
Upto 2 v v-1 min
Based on
bioreactor
volume
0.1%
pH
pH electrode
2-12
0.02
pO2
Polarografc
electrode
0-400 mbar
2 mbar
pCO2
Membrane
covered pH
electrode
0-100 mbar
2 mbar
PELETAKAN SENSOR PADA
BIOREAKTOR
Application
measurement
Lower ports
Upper ports
Headplate ports
Temp./pH/DO/ DCO2/
Turbidity
probes/substrate (s)
on-line
+ (for pilot
scale
bioreactors)
-
+ (only for small scale
bioreactors / Glass
bioreactors)
Pressure gauge or
Pressure sensor
-
-
+
Line to outgas
analysis
-
-
+ (after condenser and
out-gas fleter)
Feeding / acid-based /
Antifoam addition
-
Highly suitable (Small
Highly suitable (Large
scale pilot)
scale)
Less Suitable (Large scale) Less Suitable (Small scale
pilot)
Harvest
(Continuous Culture)
(Repeated batch)
+
-
-
BIOSENSOR
BIOSENSOR
Biosensors adalah istilah untuk mengkombinasikan substrat biologis pada
rangkaian elektronik. Hal ini merupakan generasi baru untuk pengukuran
analitis interaksi biomolekular spesifk.
"...a self-contained integrated device which is capable of providing specifc
quantitative or semi-quantitative analytical information using biological
recognition element which is in direct spatial contact with a transducer
element." – IUPAC (International Union of Pure and Applied Chemistry)
Immobilize one of
interacting molecul
(Ligand) onto inert
substrate
Substrat is bounded
onto metal surface
(gold or platinum)
Convert to electricity
signal
METODE TEKNIK
TRANSDUSER BIOSENSOR
1. Potentiometric or amperometric, where a chemical
or biological
reaction
produces a potential
difference or current fow across a pair of
electrodes.
2. Enzyme thermistors, where the thermal effect
of the chemical or biological
reaction is
transduced into an electrical resistance change.
3. Optoelectronic, where a chemical or biological
reaction evokes a change in light transmission.
4. Electrochemically
sensitive
transistors
whose
PRINSIP KERJA BIOSENSOR
PROBLEM 1
Sebutkan contoh jenis BIOSENSOR
dan penerapannya untuk sistem
instrumentasi bioproses!
MASSA SEL
CELL MASS MEASUREMENT
Pengukuran massa sel antara lain dapat dilakukan dengan metode
optical density. Pengukuran jenis ini paling efektif untuk bakteri
yang memiliki bentuk spheris misal E. coli.
Metode lain yang digunakan diantaranya adalah titrasi
spektrofotometri
Optical density (OD) =
A/L
Where A refers to
absorbance
ONLINE CELL MASS
DETERMINATION METHOD
Principle
Advantages
Disadvantages
Optical density
Wide linear range
Some interference
Culture fuorescence
Measurement of cellular
activity
Singanal interpretation
is difcult
Capacitance
Wide measurement range
Interferance from
aeration and aeration
Signal interpretation is
difcult
Measurement of cellular
activity
Ultrasonic
Wide linear range
Self cleaning
Interference from
aeration and agitation
Temperature sensitive
ONLINE CELL MASS AND
VIABILITY MEASUREMENT
Theory:
Cells with intact plasma membranes
act like tiny capacitors under the
infuence of electric feld. The nonconducting
nature
of
plasma
membranes allows charge to build
up. The resulting capacitance can
be measured and is usually
expressed
in
pico-farad
per
centimeter (pF/cm). It depends on
the cell type and is directly
proportional to membrane bound
volume of viable cells.
KOMPOSISI KIMIA
PENGUKURAN KOMPOSISI
KIMIA
Metode yang paling banyak digunakan adalah
kromatograf
Gas chromatography
Liquid Chromatography
Low Pressure
High Pressure
Thin Layer
DISSOLVED OXYGEN
DO
DO merupakan salah satu parameter terpenting yang perlu diamati
dalam proses fermentasi. Parameter ini mengindikasikan potensi
kembang biak bakteri.
Dua prinsip operasi digunakan pada probe pengukuran DO yaitu:
reaksi elektrokimia dan prinsip amperometrik (polarographic).
DO-METER
PRINSIP KERJA PROBE DO
In the amperometric (polarographic) approach, oxygen again
permeates a diffusion barrier and encounters an electrochemical
cell immersed in basic aqueous solution. A potential difference of
approximately 1.3 V is maintained between the anode and cathode.
As the oxygen encounters the cathode, an electrochemical reaction
occurs:
The hydroxyl ion then travels to the anode where it completes the
electrochemical reaction process:
The concentration of oxygen is directly proportional to the amount
of current passed through the cell.
HUBUNGAN ANTARA DO DAN
TEMPERATUR
EXHAUST GAS
ANALYSIS
ANALISIS GAS EXHAUST
Banyak yang bisa dipelajari dari gas
hasil metabolisme seperti CO2, O2, N2,
NH3 dan ethanol. Faktanya analisis
prediktif didasarkan pada penghitungan
Laju uptake oksigen, laju pertukaran
karbon dioksida yang menggambarkan
proses respirasi.
Metode yang digunakan secara luas
adalah spektrometri massa. Dua metode
operasi digunakan yaitu:
Magnetic
Quadrapole
MAGNETIC
QUADRAPOLE
Menggunakan elektron beam
untuk ionisasi partikel.
The quadrapole is a set of four
similar and parallel rods (see Fig.
4) with opposite rods electrically
connected. A radio frequency and
dc charge of equal potential, but
opposite charge, is applied to each
set of the rods. By varying the
absolute potential applied to the
rods, it is possible to eliminate all
ions except those of a specifc
mass-to-energy ratio.
PENGUKURAN PH
NILAI PH UNTUK BEBERAPA
CAIRAN
PH DAN KONSENTRASI ION
HIDROGEN
PENGUKURAN PH
EFEK TEMPERATUR PADA
PENGUKURAN PH
TABEL ERROR TEMPERATUR
JURNAL –
PENGUKURAN PH FERMENTASI – NDT
ULTRASONIK
BROTH LEVEL
As the broth in a fermenter or bioreactor becomes more
viscous and is subjected to agitation from sparging (the
introduction of tiny sterilized air bubbles at the bottom of the
liquid) and from mixing by the impeller, it has a tendency to
foam. This can be a serious problem as the level may rise to
the point where it enters the exhaust gas lines clogging the
ultraflters and possibly jeopardizing the sterile environment
within the reactor. Various antifoam strategies can be
employed to correct this situation, however, detection of
the condition is frst required.
SENSOR ANTIFOAM
STANDAR
Type of foam:
Early foaming
Late foaming
Antifoam efects on:
DO
Growth morphology
(Filamentous MO)
Foam sensors:
Low foam sensor
High foam sensor
PROBLEM 2
Jelaskan mengapa strategi antifoam
diperlukan, dan jelaskan cara kerja
sensor untuk pengukurannya!
INSTRUMENTASI UNTUK
PENGUKURAN PROSES
FERMENTASI AKTIF
CONTOH DATA HASIL
PENGUKURAN
PENGGUNAAN DATA
PENGUKURAN BIOPROSES
Data hasil monitoring bioproses antara lain digunakan untuk:
1. FAULT ANALYSIS
2. PROCESS MODELING
3. STATE ESTIMATION
4. CONTROL SYSTEM
FAULT ANALYSIS
FAULT ANALYSIS
Faults in reactor operation affect 15-20% of fermentations [25].
Fermentation measurements can be used to detect faults; for example,
signals from a fow sensor could be used to detect blockages in a
pipe and trigger an alarm in the factory control room. Normally
however, the sensors themselves are the most likely components to
fail; rates of failure of some fermentation instruments are listed in
Table 13.2.
PEMODELAN
BIOPROSES
PEMODELAN BIOPROSES
Model secara umum adalah hubungan matematika dari variabel2
proses.
As an example, a frequently-used mathematical model for batch
fermentation consists of the Monod equation for growth and an
expression for rate of substrate consumption as a function of
biomass concentration:
STATE ESTIMATION
PENDUGAAN KONDISI
PROSES
Terkadang ada beberapa variabel yang tidak
dapat diukur secara langsung pada bioproses
maka diperlukan estimasi.
Often, considerable delays are involved in
off-line measurement of important variables
such as biomass, substrate and product
concentrations.
Such delays make effective
control of the reactor difcult if control action
is dependent on the value of these parameters,
but must be undertaken more quickly than
off-line analysis allows. One approach to this
problem
is to
use
available
on-line
measurements
in
conjunction
with
mathematical models of the process to
estimate unknown variables.
Program komputer yang dikembangkan untuk ini
dinamakan software sensor, estimator atau
observer.
SISTEM KONTROL
UMPAN BALIK
PEMODELAN BIOPROSES
Jenis sistem kontrol yang digunakan umumnya adalah feedback
control system.