Industrial Stoichiometry 1

Material Balance

Dr. M. Azam Saeed

Objectives:
• Understand the terms: system,
surroundings, boundary and process;

• Be able to identify open and closed

systems, and batch, semibatch, fed batch
and continuous processes;

• Understand the difference between steady

state and equilibrium;
• Be able to write appropriate equations for
conservation of mass for processes with
and without reaction;

• Be able to solve simple mass-balance

problems with and without reaction; and

• Be able to apply stoichiometric principles

for macroscopic analysis of cell growth
and product formation.
 Overview
• One of the simplest concepts in process
engineering is a material or mass balance.

• The law of conservation of mass provides

the theoretical framework for material
balances.
 Introduction
• Mass balances provide a very powerful
tool in engineering analysis.

• Many complex situations are simplified by

looking at the movement of mass and
equating what come out to what goes in.
 System and Process
• A system consists of any matter identified for
investigation.

Surroundings

System

System boundary
• The system boundary may be real and
tangible, such as wall of a beaker fermenter,
or imaginary.

• If the boundary does not allow mass to pass

from system to surrounding and vice versa,
the system is a closed system with a
constant mass.

• Conversely, a system able to exchange mass

with surroundings is an open system.
 Continuous
Batch allows matter to flow
operates in a in and out of
closed system the system

Types of Process

semi-batch fed-batch
allows either input allows input of material
or output of mass, to the system
but not both. but not output
 Batch process
In batch cultures, a bioreactor is
filled with fresh medium and then
inoculated.

At the end of the fermentation,

the contents are removed for down
stream processing
othe reactor is then cleaned
osterilized and
orefilled for the next fermentation
 Fed batch
With fed batch cultures, fresh
media is added to the
bioreactor without continuous
removal.

When the reactor volume is

full, the fermenter is emptied,
either partially or completely,
and the process restarted
Continuous process
• With continuous cultures
– fresh media is continuously added into the
bioreactor and at the same time
– bioreactor fluid is continuously removed.

• The cells thus continuously propagate on the

fresh medium entering the reactor and at
same time, products, metabolic waste
products and cells are removed in the
effluent.
• Continuous culture reactors need to be shut
down less frequently than batch systems. Cells
can also be immobilized in the reactor to
maximize their retention and thus increase
productivity.

• Microbial populations can be maintained in a

state of exponential growth over a long period
of time by using a system of continuous
culture.
 Steady state and Equilibrium
steady state

•all properties of a system, such as temperature, pressure,

concentration, volume, mass, etc do not vary with time.
•does not change with time.
• According to this definition of steady state, batch, fed
batch and semi-batch processes cannot operate under
steady state conditions.

• Mass of the system is either increasing or decreasing

with time during fed-batch and semi batch process.

• In batch processes, even though the total mass in

constant, changes occurring inside the system cause
the system properties to vary with time.

• Such processes are called transient or unsteady-state

processes.
• Continuous processes may be either steady
state or transient.

• It is usual to run continuous processes as

close to steady state as possible.

• However, unsteady-state conditions will exist

during start-up and for some time after any
change in operation conditions.
• Steady state is an important and useful
concept in engineering analysis.

• A system at equilibrium is one in which all

opposing forces are exactly counter-balanced
so that the properties of the system do not
change with time.
• From the experience we know the system
tend to approach an equilibrium condition
when they are isolated from their
surrounding.

• In equilibrium there is no net change in either

the system or the universe.

• Equilibrium implies that there is no net driving

force for change.
• The energy of the system is at minimum and
in rough terms, the system is ‘static’,
‘unmoving’ or ‘inert’.

• For example, when liquid and vapour are at

equilibrium in a closed vessel, although there
may be constant exchange of molecule
between the phases, there is no net change in
either the system or the surroundings.
 Law of Conversation of Mass
• Mass is conserved in ordinary chemical and
physical processes.

Mi Mo
System

(kg/h glucose) (kg/h glucose)

• Consider the system ,operating as a continuous

process with input and output streams containing
glucose.
• The mass flow rate of glucose into the
system is Mi kg h-1 and the mass flow rate
out Mo kg h-1. If Mi and Mo are different
there four possible explanations:

Measurement of Mi and Mo are wrong

The system has a leak allowing glucose to
enter or escape undetected
Glucose is consumed or generated by
chemical reaction within the system
Glucose accumulate within the system
• If we assume that the measurement are
correct and there are no leaks, the difference
between Mi and Mo must be due to
consumption or generation by reaction and/or
accumulation.
• A mass balance for the system can be written
in a general way to account for these
possibilities:
• Equation 1
mass in  mass out  mass  mass  mass 
through  through  generated  consumed  accumulate d 
        
  -   +   -   =  
 system   system   within   within  within 
boundaries  boundaries  system  system  system 

• The accumulation term in the above equation

can be either positive or negative.

• Negative accumulation represents depletion of

pre-existing reserves.
• Eq. (1) is known as general mass-balance
equation.

• The mass; referred in the equation; can be

total mass or mass of a particular molecular/
atomic species.
 Types of Material Balance
Continuous processes it is
usual to collect information
about the process referring
to a particular instant
in time.
Amount of mass entering
and leaving the system
are specified using a
flow rate
Can be used directly Eq. (1)
Called a differential balance.
Batch and semibatch
processes
Information about these
systems is usually collected
over a period of time rather
Two Types than at a particular instant.
Each term of the
mass-balance equation
in this case is a quantity
of mass, not a rate.
Called an integral balance.
• Two types of balances may be written:
– Differential balances, or balances that indicate
what is happening in a system at an instant in
time. Each term of the balance equation is a rate
(rate of input, rate of generation etc.) and has
units of the balanced quantity unit divided by a
time unit (people/yr, g SO2 /s, barrels/day).
– This is the type of balance usually applied to a
continuous process.
– Integral balances, or balances that describe what
happens between two instants of time. Each term
of the equation is an amount of the balanced
quantity and has the correspond in g unit (people,
g SO2, barrels).
– This type of balance is usually applied to a batch
process with the two instants of time being the
moment after the input takes place and the
moment before the product is withdrawn.
• The following rules may be used to simplify
the material balance equation:
– If the balanced quantity is total mass, set
generation = 0 and consumption = 0.
– If the balanced substance is a nonreactive species
(neither a reactant nor a product), set generation
= 0 and consumption = 0.
– If a system is at steady state, set accumulation =
0, regardless of what is being balanced. By
definition, in a steady-state system nothing can
change with time, including the amount of the
balanced quantity.
 Simplification of the General Mass-Balance
Equation
• If a continuous process is at steady state, the
accumulation term on the right-hand side
must be zero.
• This follows from the definition of steady state:
because all properties of the system, including
its mass, must be unchanging with time, a
system at steady state cannot accumulate
mass.
• Eq. (1) becomes:
 mass in + mass generated = mass out + mass
consumed (Eq. 2)
• Eq.(2) is called the general steady-state mass-
balance equation.
• Eq. (2) also applies over the entire duration of
batch and fed-batch processes» 'mass out in
this case is the total mass harvested from the
system so that at the end of the process there
is no accumulation.
• If reaction does not occur in the system, or
if the mass balance is applied to a
substance that is neither a reactant nor
product of reaction, the generation and
consumption terms in Eq (1) and (2) are
zero.

• When reaction does not occur, Eq. (2) can

be further simplified to:

mass in = mass out (3)

Example 1: General mass-balance equation
A continuous process is set up for treatment of
wastewater. Each day, 105 kg cellulose and 103 kg
bacteria enter in the feed stream, while 104 kg
cellulose and 1.5 x 104 kg bacteria leave in the
effluent. The rate of cellulose digestion by the
bacteria is 7 x 104 kg. The rate of bacteria growth is 2 x
104kg d-1; the rate of cell death by lysis is 5 x 102kgd-1.
Write balance for cellulose and bacteria in the system.
Solution
Cellulose is not generated by the process, only
consumed. Using a basis of 1 day, the cellulose
balance in kg from Eq. (1) is:
(105 – 104 + 0 - 7 x 104) = accumulation
Therefore » 2 x 104 kg cellulose accumulates in the
system each day.

Performing the same balance for bacteria:

(103 – 1.5 x 104 + 2 x 104 – 5 x 102) = accumulation
Therefore 5.5 x 103 kg bacterial accumulates in the
system each day.
Material Balances on a Continuous Distillation
Process
• One thousand kilograms per hour of a mixture of
benzene (B) and toluene (T) containing 50%
benzene by mass is separated by distillation into
two fractions. The mass flow rate of benzene in
the top stream is 450 kg B/h and that of toluene
in the bottom stream is 475 kg T/h. The
operation is at steady state. Write balances on
benzene and toluene to calculate the unknown
component flow rates in the output streams.
Mass balance for the Cells

Mass balance for the fluid

 Integral Balances on Batch Processes
• Ammonia is produced from nitrogen and
hydrogen in a batch reactor.
• At time t = 0 there are no mol of NH3 in the
reactor, and at a later time tf the reaction
terminates and the contents of the reactor,
which include nf mol of ammonia, are
withdrawn.
• Between to and tf no ammonia enters or
leaves through the reactor boundaries, so
the general balance equation (Equation 1) is
simply generation = accumulation.
• Moreover, the quantity of ammonia that
builds up (accumulates) in the reactor
between to and tf is simply nf - no, the final
amount minus the initial amount.

• The same reason may be applied to any

substance participating in a batch process to
obtain
• accumulation
= final output - initial input (by definition)
= generation - consumption ( Equation 1)
• Equating these two expressions for the
accumulation yields
initial input + generation = final output + consumption (3)

• This equation is identical to Eq. 2 for continuous

steady-state processes, except that in this case
the input and output terms denote the initial
and final amounts of the balanced substance
rather than flow rates of the balanced
substance in continuous feed and product
streams.
Procedure for material-balance
calculations
• The first step in material-balance calculations
is to understand the problem. Certain
information is available about a process; the
task is to calculate unknown quantities.

• Material balances should be carried out in an

organised manner; this makes the solution
easy to follow
 The guidelines
• Draw a clear process flow diagram showing
all relevant information.
A simple box diagram showing all streams
entering or leaving the system allows
information about a process to be
organised and summarised in a convenient
way. All given quantitative information
should be shown on the diagram.
• Select a set of units and state it clearly.

• Select a basis for the calculation and state in

clearly.
In approaching mass-balance problem it is helpful
to focus on a specific quantity of material entering
or leaving the system.
Continuous processes at steady state we usually
base the calculation on the amount of material
entering or leaving the system within a specified
period of time.
 Batch or semi-batch processes, it is
convenient to use either the total amount
of material fed to the system or the
amount withdrawn at the end.

• State all assumptions applied to the

problem.
• Identify which components of the system, if
any, are involved in reaction.
This is necessary for determining which mass
balance equation (2) or (3) is appropriate. The
simpler Eq. (3) can be applied to molecular
species which are neither reactants nor
products of reaction.
Example Setting up a flow sheet.
Humid air enriched with oxygen is prepared
for a gluconic acid fermentation. The air is
prepared in a special humidifying chamber. 1.5
l h-1 liquid water enters the chamber at the
same time as dry air and 15 gmol min-1 dry
oxygen gas. All the water is evaporated. The
outflowing gas is found to contain 1 % (w/w)
water. Draw and label the flow sheet for this
process.
Solution:
Let us choose units of g and min for this
process; the information provided is first
converted to mass flow rates in these units.
The density of water is taken to be 103 g l-1 ;
therefore:
Unknown flow rates are represented with
symbols. As shown in Figure, the flow rate of
dry air is denoted D g min-1 and the flow rate
of humid, oxygen-rich air is H g min-1. The
water content in the humid air is shown as 1
mass%.
Stoichiometry of growth and product
formation
• The law of conservation of mass has been
used to determine unknown quantities
entering or leaving processes.

• For mass balances with reaction, the

stoichiometry of conversion must be known
before the mass balance can be solved.
• When cell growth occurs, cells are a
product of reaction and must be
represented in the reaction equation.

• In this section we will discuss how

reaction equations for growth and product
synthesis are formulated.
Growth Stoichiometry and Elemental Balances

• Despite its complexity and the thousands of

intracellular reactions involved, cell growth
obeys the law of conservation of matter.

• All atoms of carbon, hydrogen, oxygen,

nitrogen and other elements consumed during
growth are incorporated into new cells or
excreted as products.
• Confining our attention to those compounds
taken up or produced in significant quantity, if
the only extracellular products formed are CO2
and H2O, we can write the following equation
for aerobic cell growth:

CwHxOyNz + aO2 + bHgOhNi cCHαOβNδ + dCO2 + eH2O

(Eq. 4)
• In Eq. 4,
CwHxOyNz is the chemical formula for the
substrate (e.g. for glucose w= 6, x= 12, y= 6
and z= 0)
HgOhNi is the chemical formula for the nitrogen
source
CHαOβNδ is the chemical 'formula' for dry
biomass.
a, b, c, d and e are stoichiometric coefficients.
• The equation is written on the basis of one
mole of substrate; therefore a moles O2 are
consumed and d moles CO2 are formed per
mole substrate reacted, etc.

• As illustrated in Figure, the equation

represents a macroscopic view of metabolism;
it ignores the detailed structure of the system
and considers only those components which
have net interchange with the environment.
Figure 1: Conversion of substrate, oxygen and
nitrogen for cell growth
• In Eq. (4), biomass is represented by the
formula CwHxOyNz

• There is no fundamental objection to having a

molecular formula for cells, even if it is not
widely applied in biology.

• The formula is a reflection of the biomass

composition.
• As shown in Table 1, microorganisms such as
Escherichia coli contain a wide range of
elements; however 90-95% of biomass can be
accounted for by four major elements: C, H, O
and N.

• Eq. (4) is not complete unless the

stoichiometric coefficients a, b, c, d and e are
known.
Table 1 Elemental composition of Escherichia
coli bacteria
• Once a formula for biomass is obtained, these
coefficients can be evaluated using normal
procedures for balancing equations, i.e.
elemental balances and solution of
simultaneous equations.

• C balance: w =c+d (5)

• H balance: x + bg =cα+2e (6)
• O balance: y + 2a+bh =cβ +2d+e (7)
• N balance: z + bi = cδ (8)
• Notice that we have five unknown coefficients
(a, b, c, d and e) but only four balance
equations. This means that additional
information is required before the equations
can be solved. Usually this information is
obtained from experiments. A useful
measurable parameter is the respiratory
quotient (RQ):
 moles CO 2 produced d
Respirator y Quotient, RQ = =
moles O 2 consumed a

• When an experimental value of RQ is

available, Eqs (5) to (9) can be solved to
determine the stoichiometric coefficients.

• The results, however, are sensitive to

small errors in RQ, which must be
measured very accurately.
• When Eq. (4) is completed, the quantities of
substrate, nitrogen and oxygen required for
production of biomass can be determined
directly.
 Example 3 Stoichiometric coefficients for cel growth

Production of single-cell protein from hexadecane is

described by the following reaction equation:

C16H34 + aO2 + bNH3 cCH1.66O0.27N0.20 + dCO2 + eH2O

where CH1.66O0.27N0.20 represents the biomass.

If respiratory quotient,RQ = 0.43, determine the
stoichiometric coefficients.
C balance: 16 = c+ d (1)

H balance: 34+3 b=1.66c+2e (2)

O balance: 2 a=0.27 c+ 2 d+ e (3)

N balance: b=0.20 c (4)

RQ: 0.43 = d/a (5)

b is already written simply as a function of c in
(4); let us try expressing the other variables
solely in terms of c. From (1):

d= 16- c. (6)

From (5):
d
a = = 2.326 d .( 7 )
0.43
Combining (6) and (7) gives an expression
for a in terms of c only:
a = 2.326 (16 -c)
a = 37.22 - 2.326 c (8)

Substituting (4) into (2) gives:

34 + 3 (0.20 c) = 1.66 c+ 2 e
34 = 1.06 c+ 2 e
e=17 - 0.53c
Substituting (8), (6) and (9) into (3) gives:

2 (37.22 - 2.326 c) = 0.27 c+ 2 (16 - c) + (17 -

0.53 c) 25.44 = 2.39 c
c= 10.64

Using this result for c in (8), (4), (6) and (9) gives:

a = 12.48
b = 2.13
d = 5.37
e = 11.36
Check that these coefficient values satisfy Eq.
(1)-(5). The complete reaction equation is:

C16H34 + 12.5O2 + 2.13NH3

10.6CH1.66O0.27N0.20 +5.37CO2 + 11.4H2O
Recycle, purges and bypasses
• We have performed mass balances on
simple single-unit processes. However,
steady-state systems incorporating recycle,
by-pass and purge streams are common in
bioprocess industries.
Figure 2: Flow sheet for process with
(a) recycle
(b) by-pass and
(c) purge streams
• As an example, consider the system use cells as
catalysts in fermentation processes.

• It is often advantageous to recycle biomass from

spent fermentation broth.

• Cell recycle requires a separation device, such as

a centrifuge or gravity settling tank, to provide a
concentrated recycle stream under aseptic
conditions.
Figure 3: Fermenter with recycle
Figure 4: System boundaries for cell-
recycle system
• System I represents the overall recycle
process; only the fresh feed and final product
streams cross this system boundary.

• In addition, separate material balances can be

performed over each process unit: the mixer,
the fermenter and the settler.
• Other system boundaries could also be
defined; for example, we could group the
mixer and fermenter, or settler and fermenter,
together.

• Material balances with recycle involve

carrying out individual mass-balance
calculations for each designated system.
Problem
Sub systems