CHAPTER 3

RESEARCH METHODOLOGY

3.1 Introduction

This section focuses on types of chemicals, materials, equipment and procedures

involved to investigate the effect of temperature and catalyst loading in epoxidation of crude
palm oil (CPO) to epoxy yield and oxirane ring stability. The steps to carry out this study is
summarized in the experimental flow chart as Figure 3.1.

3.2 Flowchart

The experimental work flowchart for this research is shown in Figure 3.1:

Design of Experiment (DoE)

Experimental Work

Analytical Techniques

Optimisation by Response
Surface Methodology (RSM)

Figure 3.1: Experimental flowchart

3.3 Chemicals and Materials
Crude Palm Oil which contains unsaturated double bond for epoxidation was purchased
from Chung Chemical Sdn Bhd with 90% purity. Aqueous hydrogen peroxide (30 w/w%) and
formic acid reagent grade (98%) for peracid formation were obtained from Fisher Company.
Meanwhile, sulphuric acid (98%) as catalyst was purchased from Fisher Scientific. For OOC
test procedure, glacial acetic acid, crystal violet indicator solution, hydrogen bromide and
potassium phthalate were purchased from Across Organics Sdn Bhd.

3.4 Equipment

For this research, there are several types of equipment used where Table 3.1 shows the
lists of equipment and their function.

Table 3.1: Lists of equipment and their function.

Equipment Function
Motored stirrer with impeller To stir the mixture continuously
Peristaltic pump To add hydrogen peroxide in dropwise manner
FTIR To confirm the presence of epoxide’s functional group

3.5 Design of Experiment (DoE)

One of the important aspect in Response Surface Methodology (RSM) is the design of
experiments (DoE). The objective of DoE is the selection of points where the response should
be evaluated [1]. A commercial software, Minitab software was used to generate the
experimental design and response surface models where the coefficients of equation were
determined [2]. A central composite design (CCD) has been chosen as the model employed.
CCD combines the original fractional factorial or full factorial design with axial runs and centre
points to produce Central Composite Design experiments [3]. CCD are first-order (2𝑁 ) designs
augmented by additional centre and axial points to allow estimation of the tuning parameters
of a second-order model. It has two variables at three different levels of a high (+1), centre
point (0) and a low (-1) [4]. Two factors including catalyst loading (X1) and temperature (X2)
to target yield of epoxy and oxirane ring stability which were measured through percentage of
oxirane oxygen content (OOC) as a function of time.
The centre points at each face of the factorial space is α = ±1. The design provided
would give high quality of predictions over the entire design space and does not required points
outside the original factor range [5]. A multiple regression analysis of the second-order
polynomial equation was generated as a general mathematical expression for the experimental
data and was described as equation below [6]:

𝑌 = 𝛽0 + ∑𝑛𝑖=1 𝛽𝑖 𝑋𝑖 + (∑𝑛𝑖=1 𝛽𝑖𝑖 𝑋𝑖 )2 + ∑𝑛𝑖<𝑗 𝛽𝑖𝑗 𝑋𝑖 𝑋𝑗 (3.1)

In which 𝛽0 ; 𝛽𝑖 ; 𝛽𝑖𝑖 and 𝛽𝑖𝑗 are constant, linear, square and interaction regression
coefficient terms respectively. Meanwhile, 𝑌 is the predicted response, 𝑋𝑖 and 𝑋𝑗 are the
independent variables [6]. In this study, the n-value was 2, so equation (3.1) can be written as
[5]:

𝑌 = 𝛽0 + 𝛽1𝑋1 + 𝛽2 𝑋2 + 𝛽12 𝑋1 𝑋2 + 𝛽11 𝑋12 + 𝛽22 𝑋22 (3.2)

Table 3.2: Codes and actual levels of the independent variables for design of experiment.

Independent variables Symbols Coded levels

-1 0 +1
Catalyst Loading (g) X1 0 0.5 1
Temperature (°C) X2 45 65 85

In order to evaluate the pure error in the experiment, a total of 13 experiments were
amplified with replications of the centre points. The design of experiments carried out are
shown as the table below:
Table 3.3: The order and conditions for three level Central Composite Design (CCD).

Run Order Catalyst Temperature (°C)

Loading (g)
1 1.0 45
2 0.5 65
3 0.5 55
4 1.0 65
5 0.5 55
6 0.5 45
7 0.0 45
8 0.0 65
9 0.5 55
10 0.5 55
11 0.0 55
12 0.5 55
13 1.0 55
3.6 Experimental Work
The experimental work for crude palm oil epoxidation consists of experimental set-up
that portrayed the arrangement of apparatus for laboratory works and the procedure to perform
the experiment. Meanwhile, the reaction parameters for epoxidation of CPO can be seen as
Table 3.4.
Table 3.4: Reaction parameters for epoxidation of CPO.

Reaction parameters Value Units

Agitation speed 600 rpm
Catalyst loading 0-1 gram
Temperature 45-65 °C
Formic acid to unsaturated double bond molar 2 :1 -
ratio
Hydrogen peroxide to unsaturated double bond 4:1 -
molar ratio

3.6.1 Experimental Set-up

Figure 3.2: Experimental Set-up for epoxidation of CPO.

3.6.2 Experimental Procedure
The epoxidation of palm oil with performic acid generated in-situ was carried out in a
500 ml beaker equipped with motored stirrer with low shear impeller to ensure complete
mixing, thermometer, and water bath [7]. The required amount of CPO was placed in the
beaker. The calculated amounts of sulphuric acid and formic acid added. The mixture of the
CPO and formic acid were heated to the reaction temperature by controlling the temperature of
hot plate [2] and continuously stirred at 600 rpm. Then, aqueous hydrogen peroxide was added
dropwise by peristaltic pump [8]. Fast introduction to this reagent is not recommended as the
reaction will cause an excessive development of oxygen due to the decomposition of hydrogen
peroxide at high temperature [9].

Then, samples are taken every 5 minutes interval for Oxirane Oxygen Content (OOC)
analysis. Once the OOC reached maximum value, the heating is stopped. The epoxidation
reaction was repeated with different parameters of temperature and catalyst loading. The
sample are analysed by using FTIR to detect the presence of epoxide’s functional group.

3.7 Analytical Techniques

The sample was retrieved every 5 minutes to determine the experimental oxirane
oxygen content (OOC) that eventually shows the percentage conversion of unsaturated double
bond to epoxide. Then, the sample was tested for its functionality group by using FTIR
spectrum.

3.7.1 Oxirane Oxygen Content (OOC) Analysis

The product which is epoxide was analysed by using wet analytical method to
determine the percentage of experimental oxirane oxygen content (OOCexp) by using direct
method Cd 9-57 as proposed by American Oil Chemists’ Society (1997). It is the standard
method to measure oxirane oxygen which is the oxygen contained either in epoxidized fatty
materials or epoxy compounds in general. Under prescribed conditions of this method, the
oxygen is titrated directly with hydrogen bromide in acetic acid [10]. The procedure to perform
OOC analysis can be referred in Appendix A. The experimental oxirane oxygen content can be
calculated as in equation 3.1:

𝑇 𝑥 𝑁𝐻𝐵𝑟 𝑥 1.6
OOC exp = (3.1)
𝑊
 where T is the volume of HBr solution used for titration (in mL), N is normality of HBr
and W is the weight of the sample (in gram). The iodine value (IV) of palm oil used which
indicates the degree of unsaturation of palm oil was taken based on the average value of IV
determined in previous studies. Table 3.5 shows summary of IV determined from previous
study.

Table 3.5: Iodine value determined from previous studies.

Tittle of Journals IV Value

Comparison of Iodine Values (IV) of crude palm oil from 53.4560
GLC calculations with those from Wus method [11].
Determination of Iodine Value of palm oil 54.0440
by differential scanning calorimetry [12].
Epoxy resins modified with palm oil derivatives - 51.0300
preparation and properties [13].

53.4560 + 54.0440 + 51.0300

Average Iodine Value (IV) =
3

Average IV = 52.8433

The iodine value then applied to calculate theoretical oxirane oxygen content (OOCtheo) as in
equation 3.2.

𝑜(𝐼𝑉 /2𝐴 )
𝐼
OOC theo = [100 + (𝐼𝑉 ]𝐴𝑜 𝑥 100 (3.2)
/2𝐴 )𝐴
𝑜 𝐼 𝑜

where OOCtheo is the theoretical maximum oxirane oxygen, IVo is the initial iodine value of
crude palm oil, Ao is the molar mass of oxygen and AI is the molar mass of iodine. Then, the
percentage of relative conversion to oxirane is calculated using equation 3.3 [7].

𝑂𝑂𝐶 𝑒𝑥𝑝
Relative Conversion Oxirane (% RCO) = 𝑥 100% (3.3)
𝑂𝑂𝐶 𝑡ℎ𝑒𝑜

The graph of % RCO against reaction time were plotted and analysed. The example of
the graph obtained can be seen in figure 3.3 [14].
 Figure 3.3: Graph of % RCO versus reaction time.

3.7.2 FTIR spectrum

Fourier Transform Infrared Spectroscopy (FTIR) was used to observe the functional
groups of epoxidized samples. The analysis was done twice for the epoxide samples before and
after optimization. The presence of epoxidized palm oil can be observed based on the
absorption band signal. The observation of epoxide signal band confirms the formation of
oxirane ring in the epoxidized palm oil. The typical infrared spectrum of functional groups
corresponding to their absorption range is shown in table 3.6 [15].

Table 3.6: Characteristic absorption for functional group based on FTIR analysis.

Functional group Characteristic Absorption (cm-1)

Alcohol 3200-3600
Carbon-carbon double bond 1620-1680
Carboxylic acid 1670-1820
Epoxy / oxirane ring 1210-1320
3.8 Response Surface Methodology (RSM) Optimization
RSM was applied to study the effect of catalyst loading and temperature on the
percentage of oxirane oxygen, which are identified as responses. The application of RSM not
only reduces the experimental costs and time but also recognizes the optimal process
parameters which reduces the variability in responses [3]. RSM basically combines statistical
and experimental method with data-fitting techniques [3]. The model relates the objective
functions and a group of test factors. Analysis of Variance (ANOVA) is used as multivariate
techniques to determine the optimum reaction conditions of epoxidation [1].

The significance of model was decided based on the P-value. The P-value that is less
than 0.05 indicates that the model is significant. The lower the P-value means that the variable
is more significant [6]. Other than that, through P-value, the significance of interaction among
relation variables can also be determined. Other way to show the significance interaction
among variables are through the 3-D response surfaces. Furthermore, the precision of model
can be evaluated by the regression coefficient (R2). Regression coefficient denotes the accuracy
and good ability of the polynomial model. The R2 must be in the range of 0 to 1. The closer the
value to one indicates a better statistical model. As an example, if the R 2 value is 0.9923, it
signifies that the 99.23% of the model behaviour can be interpreted for optimum condition and
only 0.77% of the full variance cannot be explained by the model [16].

Meanwhile, the adjusted coefficient of determination (adj R2) value indicates the
goodness of the model [17]. The value of adj R2 should be closed to the predicted R2 value so
that reasonable agreement and prominent correlation between the remarked values and
predicted data can be proven [17]. The regression model provides a comprehensive explanation
of the relationship between the independent process variables and the response process
variables. Lastly, the model was assessed through lack-of-fit test. It is used to compare the
variation due to the changes in combination of variable levels with the variation due to the
random errors inherent to the measurement of the generated responses [4]. The P-value of lack
of fit less than 0.05 indicates that the lack of fit was considerably significant compared to the
pure error and shows that the models predicted for the responses were adequate.

The optimum conditions chosen were tested in order to confirm the model prediction.
The experiment was carried out according to the optimum condition provided and the responses
obtained was compared with the predicted values from the model.
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