The chemical equation of the esterification process was as shown:

CH3COOH (A) + CH3 CH2CH2CH2OH (B)  CH3CH2CH2COOCH2CH3 (C) + H2O (D)

Assumptions:
1. Acetic acid is chosen as a limiting reactant.

2. This is an elementary, irreversible reaction in a batch reactor

Design Equation:
𝑑𝑋
CAO 𝑑𝑡 = -rA
Rate law:
-rA = kCACB
Stoichiometry:
CA = CAo(1-X)
CB = CAo(1-X)
Combine:
𝑑𝑋
CAO 𝑑𝑡 = kCAo2(1-X)2

𝑋 1 𝑡
∫0 dX = ∫0 𝑘𝐶𝐴𝑂 dt
(1−𝑋)2

𝑋
= kCAo t
1−𝑋

Y= mX + C

𝑋
∴ A graph of 1−𝑋 versus t is plotted based on the linear equation derive from the algorithm above to get
the value of the reaction rate constant, k.
3.3 DISCUSSIONS

The purpose of this experiment is to study the relationship of esterification of butanol and acetic acid
to form ethyl butyrate and water with the present of immobilized lipase and the temperature on the
enzymatic synthesis. The lipase activity is measured with different temperature.

According to Figure 1, at both temperature which are 27oC and 37oC show that the volume of NaOH
required to turn the phenolphthalein into pink will decrease with the time. This can be explained by
decrement of immobilized lipase existed in the mixture of acetic acid and butanol. Besides, the amount
of NaOH added decreases because at the beginning of reaction, the acetic acid is not fully become ester
which is ethyl butyrate and it is become constant after some time, that mean the esterification almost
complete.

Based on Figure 2, we can conclude that at 37℃ the conversion of reactant at a specific time is higher
than that at 27℃. As the higher of the reaction temperature, the higher kinetic energy of reactant
molecule gained which then increase the frequency of collision of reactant molecule and thus increase
the conversion of reactants at a specific time. Therefore, the volume of NaOH required to be titrated
with acetic acid at 37℃ is less at the 6th minute compare to temperature 27℃, which corresponds to
the conversion of reactant at the specific time. The maximum conversion for this esterification process
at 27℃ is 40% and at 37℃ is 48% respectively. The conversion is relatively low probably due to the
assumption that the esterification reaction is irreversible which in fact it is a reversible process which
involves the equilibrium constant in the rate law. Furthermore, the reaction rate constant obtained at
37℃ (k = 0.1542 L/mol.min) is higher than at 27℃ (k = 0.1302L/mol.min) which obey the theory of
Arrhenius equation which states that the reaction rate constant will increase as the temperature increase.
The optimum temperature for the lipase activity is about 37℃ (Mulalee.S et al, 2013) which fulfil the
result of this experiment whereby the conversion is higher at 37℃ compared to 27℃.
3.3.1 OFFSET TO ELIMINATE MASS TRANSFER EFFECT OF THE IMMOBILIZED
LIPASE IN THE REACTION KINETIC STUDY

The offset to eliminate the mass transfer effect of the immobilized lipase in the reaction kinetic study
is by using sufficient high stirring speed and small size of heterogeneous immobilized lipase. The speed
of the magnetic stirrer was set at 5 rpm and it is sufficiently high as the volume of mixing solution
involved is very small (100 mL) and the amount immobilized lipase used is only 0.6 g. According to
Fogler (2014), the mass transfer coefficient, kc is inversely proportional to the square root of particle
size, dp1/2. In order to have a reaction-limited reaction, where specific reaction rate must be very small
with respect to mass transfer coefficient (kr <<< kc), hence the particle size must be small enough to
achieve high mass transfer coefficient. Therefore, the immobilized lipase size must be small enough.

3.3.2 DETERMINATION OF LIMITING REACTANT

Limiting reactant is a reactant which is fully consumed first compared to other reactants.In our
experiment ,acetic acid is choose to become limiting reactant because immobilized lipase will be
denatured when excessive acetic acid present as the optimum pH of lipase is 7.5.

There are two ways to determine the limiting reagent. First method is to find and compare the
mole ratio of the reactants used in the reaction (approach 1). Second method is to calculate the grams
of products produced from the given quantities of reactants; the reactant that produces the smallest
amount of product is the limiting reagent (approach 2). (Staley, Dennis. (2007)

Approach 1
Find the limiting reagent by looking at the
number of moles of each reactant.
1. Determine the balanced chemical equation
for the chemical reaction.
2. Convert all given information into moles
(most likely, through the use of molar
mass as a conversion factor).
3. Calculate the mole ratio from the given
information. Compare the calculated ratio
to the actual ratio.
4. Use the amount of limiting reactant to
calculate the amount of product produced.
5. If necessary, calculate how much is left in
excess of the non-limiting reagent.
Approach 2
comparing the amount of product each
1. Balance the chemical equation for the
chemical reaction.
2. Convert the given information into moles.
3. Use stoichiometry for each individual
reactant to find the mass of product
produced.
4. The reactant that produces a lesser
amount of product is the limiting reagent.
5. The reactant that produces a larger
amount of product is the excess reagent.
6. To find the amount of remaining excess
reactant, subtract the mass of excess
reagent consumed from the total mass of
excess reagent given.
3.3.3 RATE LAW USED TO ELABORATE THE KINETICS OF ESTERIFICATION
REACTION

Le Chatelier's principle or "The Equilibrium Law” can be used to elaborate the

kinetics of esterification reaction and predict the effect of a change in conditions on a
chemical equilibrium.
When any system at equilibrium is subjected to change in concentration, temperature,
volume, or pressure, then the system read just itself to (partially) counteract the effect of the
applied change and a new equilibrium is established. (Petrucci, R., Harwood, W., Herring, F.,
Madura, J. (1993)) The yield of ester may be increased by increasing the concentration of
either reactant. Since the reaction is reversible, the simultaneous removal of product would
shift the reaction in forward direction. (M.A. Suryawanshi, N.H. Shinde, R.V. Nagotkar.
(2014))