Synthesis and Refinement Process of Base Catalyzed Biodiesel

by

Michael Witkowski

A Research Paper
Submitted in Partial Fulfillment of the
Requirements for the
Bachelor of Science Degree
in
Biochemistry

Under the direction of

Dr. Christina Edwards

_____________________________ _____________________________
Dr. Christina Edwards Dr. Adam Moser

Loras College
May 2019
 ii

Division of Molecular, Life, and Health Sciences

Chemistry/Biochemistry Program
Loras College
Dubuque, IA

Author: Michael Witkowski

Title: Synthesis and Refinement Process of Base Catalyzed
Biodiesel
Degree/ Major: BS Biochemistry
Research Adviser: Dr. Christina Edwards
Month/Year: May 2019
Number of Pages: 29

Abstract

Fossil fuels are projected to diminish completely in the next 50 to 100 years. An alternative

fuel that could replace fossil fuels is biodiesel, which is environmentally friendly and a

renewable resource. However, the refinement costs are what make it more expensive to

purchase than fossil fuels. This study focuses on what effect impurities have on the

properties of biodiesel and how the properties change when different refinement methods

are performed, such as wet-washing and dry-washing. Characterization tests such as FTIR,

NMR, density, flashpoint, viscosity, and heat of combustion were done on crude, refined,

and commercial grade (B100) samples. When looking at the FTIR and NMR spectrums,

the FTIR showed a small OH peak, while the NMR showed no methanol shift. The results

for the crude sample for density, flashpoint, and viscosity show impurities were having an

effect on the properties, but the type of impurity(s) was unknown. With the data collected

from this study, future work can be done to show how different impurities effect certain

properties of the biodiesel. This would not only help us to better understand what role

impurities are playing, but also lead us closer to improving the refinement methods.
 iii

Acknowledgements

I would like to thank Dr. Christina Edwards for her guidance during this research. I

would also like to thank Dr. Adam Moser and Dr. David Oostendorp for assisting in the

writing process for this research. Finally, I would like to thank Loras College Science

program for providing me with the funds to work on my research and Clarke University,

especially Joe Tubbs, for allowing me to use their NMR and Bomb Calorimeter.
 iv

Table of Contents

....................................................................................................................................... Page

Abstract .............................................................................................................................. ii

List of Figures ..................................................................................................................... v

List of Tables .................................................................................................................... vi

Introduction ........................................................................................................................ 1

Previous Research .............................................................................................................. 8

Methods ........................................................................................................................... 10

Results/Discussion ........................................................................................................... 13

Conclusion ........................................................................................................................ 18

References ......................................................................................................................... 19

Appendix A ...................................................................................................................... 21

Appendix B ...................................................................................................................... 25
 v

List of Figures

Figure 1. The History and Projections of the Usage of Fossil Fuels until 2040 ................ 1

Figure 2. The Comparison of the Broken Cycle and Closed Cycle .................................... 2

Figure 3. Transesterification Reaction ................................................................................ 5

Figure 4. Wet-Washing Method of Biodiesel ..................................................................... 6

Figure 5. FTIR Spectra of Vegetable Oil .......................................................................... 21

Figure 6. FTIR Spectra of B100 Biodiesel ....................................................................... 22

Figure 7. FTIR Spectra of Crude Biodiesel ...................................................................... 22

Figure 8. FTIR Spectra of Wet-Washed Biodiesel ........................................................... 23

Figure 9. FTIR Spectra of Dry-Washed Biodiesel ........................................................... 23

Figure 10. FTIR Spectra of Methanol ............................................................................... 24

Figure 11. FTIR Spectra of Glycerol ................................................................................ 24

Figure 12. NMR Spectra of B100 Biodiesel ..................................................................... 26

Figure 13. NMR Spectra of Crude Biodiesel .................................................................... 26

Figure 14. NMR Spectra of Vegetable Oil ....................................................................... 27

Figure 15. NMR Spectra of B100 Biodiesel with Methanol ............................................ 27

Figure 16. NMR Spectra of B100 with Glycerol .............................................................. 28

Figure 17. NMR Spectra of Wet-Washed Biodiesel......................................................... 28

Figure 18. NMR Spectra of Dry-Washed Biodiesel ......................................................... 29

 vi

List of Tables

Table 1. Emissions for Diesel Compared to Biodiesel ....................................................... 3

Table 2. Characteristic Regions of Vegetable Oil and Biodiesel ..................................... 13

Table 3. Measured Densities ............................................................................................. 14

Table 4. Measured Flashpoints ......................................................................................... 15

Table 5. Measured Viscosities .......................................................................................... 16

Table 6. Measured Heat of Combustion Values .............................................................. 16

 1

Introduction

Our society, today, is still very reliant on the use of fossil fuels for energy. Fossil
fuels, such as coal, oil, and natural gas, all consist of animal and plant deposits that decayed
into this organic matter over millions of years1. They are first obtained by drilling down
into the earth and then brought to the surface to be refined down so they can be used
commercially. The
use of fossil fuels does
not look like it will be
slowing down as
indicated by the
predictions in Figure
1. The estimated
consumption of the
three most used fossil
Figure 1: The History and Projections of the Usage of Fossil Fuels until
20402. fuels are on the rise.
The issue with using fossil fuels is they are very harmful to the environment and a
nonrenewable resource. The byproducts associated with fossil fuels when combusted
include: sulfur oxides, nitric oxides, carbon monoxide, carbon dioxide, and volatile organic
compounds3. The sulfuric and nitric oxides are main contributors to acid rain, which are
destroying many forested areas. Both carbon monoxide and carbon dioxide are contributing
to warmer climates because they are trapping heat from the sun inside the atmosphere,
warming the planet. Furthermore, fossil fuels are a nonrenewable resource, so they cannot
be reused after they have been burned. This is why fossil fuels have what is called a broken
cycle (Fig. 2)4. Once fuel is extracted from the ground, it is brought to a refining plant so
the impurities can be washed from the fuel. After refinement, the fuel is bought by the
costumers and used to power their vehicles or any other fuel powered device. This is where
the circle ends. Once the fuel is burned, the contaminants are ejected into the atmosphere,
and are unable to be used again in our lifetime. The reason the plants cannot use up the
carbon dioxide is because burning diesel fuel releases 161.3 tons of carbon dioxide per year
 2

into the atmosphere alone which is more than every plant on earth can handle24. At this
rate, it is predicted the oil reserves will be depleted in the next 50-100 years and the current
oil reserve sites are becoming more unstable5. The efforts in finding an alternative resource
have increased and are becoming more popular. One of these potential renewable resources
is biodiesel.

Figure 2. The Comparison of the Broken Cycle from Fossil Fuels to the Closed Circle from Biofuels 4.

Biodiesel is a renewable, biodegradable fuel manufactured from vegetable oils,

animal fats, or recycled restaurant grease6. Unlike fossil fuel with the broken circle,
biodiesel has a closed circle (Fig. 2)4. Biodiesel starts off as a plant, like a soybean or
sunflower plant. The plant is then harvested and brought to a refinery plant to be made into
biodiesel. The major byproduct that is released when it is burned, is carbon dioxide, which
plants use to grow7. The amount of carbon dioxide that is released from biodiesel is reduced
by 74% compared to the amount released by fossil fuels25. Having this closed circle is what
makes biodiesel more beneficial for the environment. It shows that the plants can keep up
with the release of CO2 produced, which will help maintain the CO2 concentration in the
atmosphere. Another benefit of biodiesel is that it is biodegradable. It takes roughly 28
days for 90-98% of biodiesel to completely degrade in soil and water compared to fossil
fuels in the same amount of time, which only degrade up to 50-56%8. This makes the fuel
much safer for the environment when a spill occurs. It is less of a risk for the wildlife when
the biodiesel decomposes a lot faster than fossil fuels.
Biodiesel is beneficial for three more reasons. The production process is simple
because all it requires are vegetable oils, which are easily made from crops, and methanol.
This makes obtaining biodiesel very easy and can be made in mass amounts. While being
 3

renewable, other advantages of biodiesel pertain to its properties which make it better than
fossil fuels. Biodiesel has a high flashpoint. A flashpoint is the temperature at which the
vapors of a material can be easily ignited. Having a high flashpoint is better for storage of
the material. Biodiesel’s flash point was tested to be over 130 oC, while fossil fuels, more
specifically diesel, has a tested flashpoint of 55oC26,4. Biodiesel is therefore safer to store
than diesel.
Finally, biodiesel emissions are reduced from that of fossil fuels. When burned, the
carbon dioxide emitted is considered carbon neutral. This means that the amount of carbon
dioxide that is emitted
into the air is the same
amount that can be
absorbed by plants10. As
for the sulfur dioxide, the
amount that gets emitted
is so low that their levels
can’t be measured10,17. In
Table 1. Emissions of Diesel Compared to Biodiesel.17 Table 1, the only harmful
emission that biodiesel emits more of is the nitrates, and even then, it is only 5% more17.
When looking at all these advantages of biodiesel over fossil fuels, it looks to be the clear
renewable, biodegradable replacement.
Biodiesel does come with some disadvantages to fossil fuels. The big issue with
biodiesel is that it is more expensive to produce than fossil fuels. In the US, the average
price to pay for one gallon of biodiesel was $3.38, while diesel was $2.76 per gallon in
201811. This is largely due to refinement costs, which make up 90% of the running costs12.
Another disadvantage is that biodiesel has a high viscosity. This means that the fuel does
not flow very well. The fuel will then require a stronger fuel pump to push it through the
tubing to the engine. One last disadvantage is that biodiesel has a low heat of combustion.
This is an issue because the amount of energy that is produced from the fuel is lower
compared to the amount from fossil fuels. While biodiesel itself doesn’t produce a lot of
carbon emission, the refinement processes that it goes through produces more waste, like
contaminated water, than that of fossil fuels12.
 4

Looking at where fossil fuels are heading in the near future and the impact they are
having on our planet, biofuels have a lot of promise. With their advantages of producing
less harmful gases and being a renewable energy source, the cut down on fossil fuels can
come to fruition. The only hurdles to overcome are cutting down on refinement costs,
lowering the viscosity, and being able to produce a higher heat of combustion. Figuring
out how to accomplish those goals, the environment will be a lot healthier and fuel would
be cheaper to obtain than fossil fuels.
 5

Background
Biodiesel is synthesized from some sort of plant based oil, methanol, a base
catalyst, and heat. In the biofuel industry, the most widely used types of oils that companies

Figure 3. Glycerol Production via Triacylglyceride Transesterification with Methanol or Ethanol. 14

use are soybean (vegetable oil), rapeseed, and canola13. The main reason these are used is
because they are mass produced by farms, with soybeans producing 1.5 gallons of oil per
bushel13. The process that turns these oils into biodiesel is called a transesterification (Fig.
3) 14. The transesterification takes the triacylglyercides found in the oils and reacts them
with methanol and a base catalyst. The base catalyst removes the hydrogen from the alcohol
group on the methanol. The negatively charged methanol will make a nucleophilic attack
on one of the fatty acid groups. This is done leaving the carbon backbone with one
negatively charged oxygen. Since the base is a catalyst, it will return the hydrogens it took
from the methanol and give them to the oxygen. The reaction happens again in a stepwise
function to remove the next fatty acid group. When the reaction is completed, the products
are biodiesel and glycerol17.
While the biodiesel is made, there are many impurities that are formed in this
reaction that have to be removed if the fuel is to be sold commercially. Some those
impurities are glycerols, free fatty acids, metals, and unsaturates15. Glycerol, which is one
of the products of the transesterification reaction, causes thermal instability in the fuel15.
Thermal instability causes the fuel to be unstable at lower than standard temperatures.
Another impurity that is formed are free fatty acids (FFAs), which come about when the
fatty acids on the triacylglyceride come off and don’t react with the methanol. These FFAs,
like glycerol, cause thermal instability as well15. The next impurity are metals, which come
from the base catalyst. The catalyst is the negatively charged alcohol, which needs a
 6

positive counter-ion, like potassium or sodium ions, to be stable. These ions are metals and
can potential be found in the biodiesel. When metals are present in the fuel, they can clump
up on the walls of the fuel lines, inhibiting the flow of fuel into the engine15. The last
impurity is unsaturates, which are formed when the triacylglyceride do not react fully with
the methanol. There would then be a combination of alcohol and fatty acid groups
connected to the carbon backbone of the triacylglyceride and they would inhibit the flow
of fuel15.
With all of these impurities inhibiting the performance of biodiesel, the United
States made specific standards fuels must abide. The standards that are of importance to
this research are flash point, density,
and kinematic viscosity. The flash point
temperature that biodiesel must achieve
is between 200-275oC16. This would
ensure the biodiesel can be safely stored
and help avoid any accidental fires. For
density, the fuel must be between 860-
900 kg/m3, so the fuel can be properly
and efficiently injected in an engine16.
Finally, the standard set for the
kinematic viscosity is between 1.9-6.0
mm2/s 16
. A kinematic viscosity is a
measure of how well the fuel can flow
through a tube. The range is set low
because the higher the viscosity, the
slower the fuel will move through the
Figure 4. Wet-Washing Method of Biodiesel19. tube. Having a low viscosity can ensure
the fuel can flow through an engine with ease. For companies to reach these standards,
there are different types of refinement methods they can use to purify the biodiesel.
There are two types of refinement processes companies use: wet-washing and dry-
washing. The wet-washing method is the oldest and most common for cleaning biodiesel18.
The biodiesel is continuously sprayed with water to rid the fuel of its impurities (Fig. 4)19.
 7

The water is able to purify the fuel because of its polar nature, attracting all the glycerols,
free fatty acids, metals, and unsaturates, and settles at the bottom. The density difference
between water and biodiesel allow for two layers to form with the water at the bottom (Fig.
4)19. This process is usually done several times and over a long period of time to ensure
that all the impurities are removed from the biodiesel18.
This type of method does come with some disadvantages. First, it uses a lot of water
to refine the fuel so it adds to the price of the fuel18. The amount of water that is used is the
same amount that becomes contaminated with the waste from the unrefined biodiesel12.
The usage of water and the process to filter the contaminated water is what accounts for
the first 45% of the running costs. The other 45% of the running costs is that it takes time
for this process to happen. Time is very valuable for companies so when washes take long
periods of time to do, companies are losing out on potential profits. Therefore, companies
increase the price to make up for lost time.
Dry-washing is a type of washing method that can purify biodiesel without using
any water. Dry-washing in itself has two methods, one using magnesol and another using
an ion exchange resin. Magnesol is a fine powder that is polarized21. Just like the wet-
washing method, the magnesol is added to the biodiesel, and the powder attracts all the
polar impurities found in the fuel. The powder will then settle at the bottom because of the
density difference from biodiesel. The best thing about magnesol is that it can be reused
multiple times before it goes bad21. However, there is a huge issue with using magnesol as
a purifier. The particles are so small that they cannot be filtered out without losing some
biodiesel21. The smallest filters at 1 micron can’t prevent the magnesol from getting into
the biodiesel21. The only way around this issue is to filter the sample multiple times to
ensure the magnesol is not in the refined biodiesel. Another type of dry washing method is
the ion exchange resin. This method uses a column filled with polarized resin beads, which
allow the nonpolar biodiesel to flow through, while the polar impurities stick to the resin21.
The beads are rinsed with a solvent to remove all the impurities from it and give it back its
charge to be used again21. The only disadvantage from using the resin is that it won’t work
if the biodiesel is high in methanol content. If too much methanol is used during the
synthesis, it would overwhelm the column enough to allow small amounts of methanol to
get through, and contaminate the biodiesel.
 8

Previous Research
A study done by Faccini et al. investigated dry washing in biodiesel purification.
The purpose of their study was to compare the efficiency of different absorbents used for
dry washing. The biodiesel was synthesized by mixing soybean oil with KOH and
methanol. A wet wash was done on some of the crude biodiesel. The fuel was washed three
times with distilled water while being constantly stirred for five minutes before the
heterogeneous mixture was separated. For dry-washing, they used different types of
powders like Magnesol, Silica, Amberlite BD10 Dry and Purolite PD 206 as their
absorbents. The crude biodiesel was heated to 65 oC then the absorbents were added. The
contents were stirred for 20 minutes and the absorbents were filtered out, leaving purified
biodiesel. The results that were obtained was that the Magnesol and silica were the best in
removing the glycerol compared to the wet-washing and other two absorbents. In
conclusion, using a dry wash decreased the total time of production and is environmentally
friendly. Even though they were able to successfully remove the excess glycerol, they
didn’t look at any of the other impurities involved with crude biodiesel, like methanol,
metals, water, and free fatty acids. Dry-washing method may not be a viable source of
refinement unless more research is done on the other impurities.
Additionally, a study done by Karaosmanoglu et al. investigated the refining step
of biodiesel production. This group wanted to find a way to obtain a high yield of purified
biodiesel and keep the refining costs down. In the study, they tested 3 different types of
refinement. The first process was a wet-wash using hot deionized water at temperatures 50,
65, and 80oC. The second refinement process used wet-washing with petroleum ether and
water. The last type of process uses sulfuric acid at a 1:1 ratio to neutralize the solution.
The results that were found were the water at the temperature of 50oC produced the highest
yield at 84.2%. The second method with the petroleum ether and water produced a yield at
82.6% and the final process with the 1:1 ratio of sulfuric acid produced a yield of 81.2%.
The group made the conclusion that the process using the hot water at a temperature of
50oC was the best method for the refinement of biodiesel because it produced the highest
yield. It was also found to be the cheapest of the three methods. The issue with this study
was that they didn’t test the properties. Even though they found what they were initially
asking, they don’t know the quality of the biodiesel they produced, or whether or not one
 9

of the other processes produced a better quality than the 50oC hot deionized wet-washing
method.
Past Loras student, Josh Schulte, investigated the comparison of commercial wet
wash refined, dry wash refined, and unrefined biodiesel. His goal was to produce a
refinement process that is cheaper and creates a better performing biodiesel than what is
commercially available. This goal was split into 2 sub goals, one was testing the
characteristics of refined biodiesel from wet washing and dry washing, and the other one
was determining the cost efficiency of both wet and dry washing. He synthesized the
biodiesel using peanut oil, methanol, and potassium hydroxide. He tested the
characterization of the biodiesel’s density, viscosity, flash point, heat of combustion, and
NMR. From the data, he was able to determine that the dry-washing method was more
successful than the wet wash. The viscosity and NMR spectra showed that the biodiesel
that was dry-washed had less impurities than the wet-washed biodiesel. This then supported
his original goal. The wet-washed biodiesel however had a higher heat of combustion. This
result shows that both types of wash had their benefits in what they eliminated from the
crude biodiesel. Josh was able to look at which refined biodiesel had the better
performance. What was not looked at, was what the impurities were doing to the crude
biodiesel, that inhibited its properties from fitting the ASTM standards.
Research Goals
The main goal of this research project is to show what effect impurities are having
on the properties of biodiesel and how the properties are changing when different
refinement methods are performed. Along with this main goal, a sub goal would be to
successfully refine the crude biodiesel so it can be compared to the ASTM and the B100
biodiesel. Knowing how the impurities are removed with different refinement methods and
how the impurities are influencing biodiesel properties, researchers can then come up with
an effective and cost efficient method of refinement. Finding a new more cost effective and
environmentally friendly refinement method will help get biodiesel on to the market, and
help save the environment from all the contaminants fossil fuels are dumping into the
atmosphere.
 10

Methods
Biodiesel Synthesis:
The synthesis required Hyvee vegetable oil (soybean oil), methanol, and KOH. A
KOH/methanol solution was made by mixing 35g of KOH to 1L of methanol. A sample of
55mL of the vegetable oil, with a temperature probe, was heated and maintained around
45-55oC while being stirred. Once the target range was reached, 14mL of KOH/methanol
solution was slowly added. The solution was heated/mixed for 30min, then poured into a
separatory funnel and allowed to cool for 15min. Two layers formed with the top being the
biodiesel and the bottom being glycerol, and both layers. This procedure was done 3
additional-times and samples collected.
Biodiesel Refinement:
A wet-wash method was done with 30mL of crude biodiesel and 30mL of distilled
water at 50oC. Both liquids were poured into a separatory funnel and shaken for 5 minutes
to agitate the biodiesel. The mixture was allowed to sit for 24hrs then two layers formed,
top layer being the biodiesel and bottom being the glycerol. The two layers were separated.
This procedure was done again with the newly refined biodiesel.
A dry-wash method was done with 30mL of crude biodiesel and solid Magnesol-
XL. The crude biodiesel was stirred and heated to 45oC. A 1% by weight Magnesol was
added and the mixture was heated/mixed for 2min. The mixture was allowed to sit for 24
hours and separated using 8 µm qualitative cellulose filter paper.
Characterization Tests:
A density test was done using the Mettler AE 160 analytic scale to measure mass.
A graduated cylinder was used to measure volume. This was done two additional times so
an average and standard deviation were calculated. This test was done with crude biodiesel,
wet-wash biodiesel, dry-wash biodiesel, KOH/methanol solution, vegetable oil, and
glycerol.
A flash point test was done with a small beaker, nearly full to the brim, of a sample
placed on a Barnstead Thermolyne Cimarec hot plate. A Vernier Lab Quest with
temperature probe recorded the temperature at the surface of the sample. A long neck
lighter was waved over the beaker every 10oC, starting at 150oC to 170oC, then every 5oC,
to 180oC, and every 1oC until ignition. This procedure was done 2 additional times so an
 11

average and standard deviation were calculated. This test was done for crude biodiesel,
wet-wash biodiesel, dry-wash biodiesel, and KOH/methanol solution.
A viscosity test was done with a Canon-Fenske viscometer tube, size 150. Roughly
5mL of the sample was poured into the large end of the viscometer. The viscometer was
placed in a hot water bath, held at 40oC, for 10 minutes. A rubber hose was placed over the
small end so the sample could get sucked up by vacuum filtration. The knob for the suction
apparatus was turned slightly so the sample would slowly move up the viscometer. Turning
the knob too fast created too strong of a suction and air pockets formed in the sample.
Suction was stopped after the sample halfway filled the second bulb. The rubber tube was
removed from the small end and the sample began to equilibrate back down the viscometer.
A stopwatch was used to time the moment the sample reached the first marked line until it
passed the second marked line. The time collected was multiplied by the manufacture’s
constant, 0.01377, at 40oC. This was done 2 additional times so an average and standard
deviation were calculated. This test was done with crude biodiesel, wet-wash biodiesel,
dry-wash biodiesel, vegetable oil, and glycerol.
A FTIR was taken on samples using the Brunker Alpha-E FTIR. For the computer
procedure to run the FTIR, refer to Appendix A. This test was done with crude biodiesel,
wet-wash biodiesel, dry-wash biodiesel, KOH/methanol solution, vegetable oil, and
glycerol.
A heat of combustion test was done on each sample using a Parr Adiabatic Bomb
Calorimeter. Parr USA Standard benzoic acid pellets, 6318 cal/g, were used to find the heat
capacity of the bomb. A Parr fuse wire, 1400 cal/g, roughly 15cm, was weighed out and
inserted through the slits of the bomb. The benzoic acid tablet was placed in the little dish
and put onto the holder. With tweezers, the wire was coiled and pushed down onto the
tablet so the coils made contact with it. The bomb was put together and sealed shut. The
bomb was filled up to 30atm of O2 gas. The bomb was placed into the bin, which was filled
with 1600mL of distilled water. The bin, with the bomb, was placed into the bomb
calorimeter and the leads were connected to the bomb. The bomb calorimeter was closed
up and the mixing wand and temperature probe were lowered into the bin. After the
temperature equilibrated the initial temperature was recorded and the ignite button was
pushed. After the temperature stopped rising, the final temperature was measured. The
 12

bomb was opened to make sure there was no black substances in the dish. If there was, the
reaction did not fully combust and had to be repeated over again. The mass of the remaining
nickel wire was measured. The Cbomb was found using Equation 1, each sample could be
ran using the same procedure above. The qsample variable is the specific heat of the tested
sample. The three m variables are the masses of the initial wire, final wire, and water
respectively. The 𝛥𝐻𝑤𝑖𝑟𝑒 is the enthalpy value for the wire. The cbomb variable is the heat capacity
of the bomb. The 𝛥𝑇 variable is the change in the temperature of the water. The 𝑆𝐻2 𝑂 variable is
the heat capacity of water. With this equation, the qsample could be calculated and the heat of
(1) −(𝑞𝑠𝑎𝑚𝑝𝑙𝑒 + (𝑚𝑤𝑖𝑟𝑒 𝑖𝑛𝑖𝑡𝑖𝑎𝑙 − 𝑚𝑤𝑖𝑟𝑒 𝑓𝑖𝑛𝑎𝑙 )𝛥𝐻𝑤𝑖𝑟𝑒 ) = 𝑐𝑏𝑜𝑚𝑏 𝛥𝑇 + 𝑚𝐻2 𝑂 𝑆𝐻2 𝑂 𝛥𝑇

combustion would be determined. This was done 2 additional times so an average and
standard deviation were calculated. This test was done for crude biodiesel, wet-wash
biodiesel, and dry-wash biodiesel. An F test and T test at the 95% confidence level, were
done on wet-washed and dry-washed biodiesel.
NMR was performed on each sample using the Eft-90 NMR Spectrometer at Clarke
University. CDCl3 was used as the solvent. The sample was mixed with the CDCl3 and
filled the NMR tube 1/5 full. For the series of commands that were put in the computer,
refer to Appendix B. This test was done for crude biodiesel, wet-wash biodiesel, dry-wash
biodiesel, vegetable oil, B100 biodiesel, B100 w/methanol, and B100 w/glycerol.
Limitations:
The NMR and FTIR are very quick and easy ways for identifying impurities that
can be found in each biodiesel sample. Both of these tests produced different results for the
crude biodiesel, which lead to further tests with the NMR for B100 contaminated with
methanol and another with glycerol. The methanol shift, in the NMR, was not present,
which was likely due to using a 90MHz NMR at Clarke University. The 300MHz NMR at
Loras College was currently down so the loss in resolution might be the reason for the
methanol shift not being present. The methanol shift was not present for the B100 with
glycerol, which there should have been a shift. The flashpoint method used is not how
flashpoint is supposed to be tested, so the temperatures could very in a way that makes the
crude fit the standard. There is an actual instrument that measures flashpoint, but this study
did not have access to the device. The other impurities in the crude sample might have gone
undetected, so finding a method that can locate them, would be beneficial.
 13

Results/Discussion
After the crude biodiesel was synthesized, characterization tests were performed to
analyze the biodiesel. In the FTIR spectrum, the following characteristics in Table 1 were
used to differentiate vegetable oil and biodiesel27. With the regions of the vegetable oil and
Region (cm-1) Assignment Vegetable oil Biodiesel

1425-1447 CH3 asymmetric bending Absent Present

1188-1200 O-CH3 stretching Absent Present

1075-1111 O-CH2-C asymmetric axial stretching Present Absent

1700-1800 C=O stretch Present Present

2800-3000 Symmetry CH2 stretching and the asymmetry CH3 Present Present
and CH2 stretching

Table 2. Characteristic Regions of Oil and Biodiesel Spectra Used for Quantification 27

biodiesel clearly identified, the FTIR spectra from the vegetable oil, crude biodiesel, and
B100 biodiesel could be analyzed. The spectra for vegetable oil showed its three present
assignments and both B100 and crude biodiesel spectrums showed their four present
assignments from Table 2. The spectra of vegetable oil, crude biodiesel, and B100
biodiesel, with labeled regions, can be found in Appendix A. In the crude biodiesel spectra,
there was a broad peak around 3400cm-1, which signifies an OH group. The OH group
could have been from the glycerol, residual methanol, or the catalysis, KOH, getting into
the biodiesel during separation. This was expected because the biodiesel did not go through
the refinement processes to rid it of contaminants. This was supported by the spectra
produced by the glycerol and methanol because both contained very broad OH peaks at
3320cm-1, therefore they were in the biodiesel and needed to be removed. The FTIR spectra
of the wet-washed and dry-washed biodiesel did not have the broad peak at 3320cm-1, and
was comparable to the B100 biodiesel spectra. With the FTIR data, it was possible to not
only know biodiesel was created, but also see potential contaminants be removed when
refined.
For the NMR results, the vegetable oil and crude biodiesel had differences. The
vegetable oil lacked the methyl ester shift at 3.66ppm, which is consistent with its carbon
back bone31. That structure is able to be seen in the spectra at 4.27ppm, 4.23ppm, and
4.17ppm. The spectra for the crude biodiesel has the methyl ester peak and loses the carbon
 14

backbone structure. That confirms to how biodiesel was pictured in Figure 3. Comparing
the crude and the B100 biodiesel spectrums, they both produced the sample spectra. This
caused concern because the crude biodiesel should have had a methanol shift at 3.31ppm
and a methyl ester shift at 3.66ppm28,29. In Appendix B, the spectra for crude biodiesel does
not contain the methanol shift that is expected to be there from the FTIR spectra. The NMR
used was scanning at 90 MHz, so it may have been possible the methanol shift was masked
by the methyl ester shift. A test to prove whether or not the methanol and methyl ester
peaks overlapped was performed in which B100 was purposely contaminated with
methanol. The NMR spectra of the contaminated B100 showed a large shift at 3.392ppm
which was not present in the B100 spectra. This indicated that the NMR used can
distinguish a methyl ester and methanol shift from one another. The most reasonable
answer for the crude biodiesel not having the methanol shift was that the methanol in the
crude sample was in such a small amount that the NMR could not detect it. This can be
supported by the FTIR spectra for the crude biodiesel because the broad band for the OH
was not a prominent OH peak showing that the crude biodiesel had a small amount of
methanol in it. For the wet-washed and dry-washed biodiesel spectra, they too, matched
the B100 spectra, which confirms the FTIR spectrums, showing no detectable contaminants
were present in the samples.
Characterization Tests:
The density of the crude biodiesel fell within the range of 0.860-0.900 g/mL. When
ASTM 0.860-0.900 g/mL referring to Table 3, the crude biodiesel
had a value of 0.865±0.008 g/mL, which
Crude Biodiesel 0.865 ± 0.008 g/mL
compared to the B100 biodiesel at
B100 Biodiesel 0.86 ± 0.01 g/mL
0.86±0.01 g/mL. This showed the
Wet-Wash Biodiesel 0.866 ± 0.007 g/mL densities were similar, which is one sign
Dry-Wash Biodiesel 0.886 ± 0.001 g/mL the biodiesel produced was performing
like its purified counterpart. When the
Table 3. Measured Densities with Uncertainty Values
for Crude, B100, Wet-washed, and Dry-washed biodiesel was refined by a wet-wash, the
biodiesel.
value was within the range of the ASTM and comparable to the B100 biodiesel. For the
dry wash biodiesel, the density was significantly different compared to the other samples.
The dry-wash showed no OH peak or methanol shift from the FTIR and NMR,
 15

respectively. A possible contaminant that may have resulted in the increase might be the
magnesol beads. There is a likelihood a few beads passed through the filter paper and were
not detected by the FTIR or NMR resulting in the significant difference in the data.
The flashpoint was tested on the crude biodiesel and in Table 4, the value was lower
than the ASTM and the B100 Biodiesel. Having the lower temperature meant that the crude
biodiesel was not as safe for storage compared to the B100. This brings up the OH peak
that was found in the FTIR spectra. The impurity that was causing the low flashpoint
between glycerol and methanol, is leftover ASTM 200-275oC
methanol from the synthesis reaction. This
Crude Biodiesel 182.7 ± 0.2oC
can be determined by the very low
Methanol < 20 oC
flashpoint of methanol. The glycerol was
tested for flashpoint, but would not spark a B100 Biodiesel 202. ± 0.1oC
flame, thus could not be the impurity Wet-Washed Biodiesel 202.1 ± 0.7oC
causing the low value. When the wet-
Dry-Wash Biodiesel 201. ± 1.oC
washed biodiesel was tested, the value in
Table 4 shows that it fit within the ASTM Table 4. Measured Flashpoints with Uncertainty
Values for Crude, Methanol, B100, Wet-washed,
and comparable to the B100 biodiesel. The and Dry-washed biodiesel.

dry-wash biodiesel also fit within the standards so the disappearance of the OH peak in the
FTIR spectra of the wet-washed and dry-washed biodiesel allowed the flashpoint to
increase to acceptable values. The flashpoint test on both refined biodiesel samples further
supports that the samples were refined to ASTM.
The viscosity results are displayed in Table 5. The crude biodiesel fit within the
ASTM, but was a little slower compared to the B100 biodiesel. This circles back to the
impurities in the crude biodiesel from the FTIR spectra. The slower value is likely due to
a small amount of glycerol that contaminated the biodiesel, during separation. An NMR
spectra of B100, purposely contaminated with glycerol, was analyzed and showed no
addition peaks compared to the crude and B100 spectrum. Even though glycerol was not
identified in the NMR spectra, it fits better than methanol because of the differences in
densities as well as the viscosity results. The value for glycerol was larger so if there was
a little bit present in the crude biodiesel, that may have caused the value to be slower
 16

ASTM 1.9-6.0 mm2/s compared to the B100. In the FTIR

Crude Biodiesel 4.768 ± 0.005 mm2/s spectra, this can be seen where the peak for
the OH is shallower than the OH peak of the
Glycerol 14.7 ± 0.5 mm2/s
glycerol, indicating there is not a definite
B100 Biodiesel 4.46 ± 0.07 mm2/s amount of OH in the crude biodiesel. When
Wet-Washed 4.68 ± 0.04 mm2/s the wet-wash and dry-washed biodiesel were
tested, their values in Table 5, fit within the
Dry-Washed 4.65 ± 0.03 mm2/s
ASTM, improved from the crude biodiesel,
Table 5. Measured Viscosity Values with their but a little slower than the B100 biodiesel. The
Uncertainty Values for Crude, Glycerol, B100,
Wet-washed, and Dry-washed Biodiesel. improvement was from the successful removal
of the impurities but still does not explain why it was slower than the B100 biodiesel. The
FTIR show that the OH peaks were gone so glycerol or methanol cannot be affecting the
value. This than brings up if there are any other impurities that might be affecting the crude
biodiesel, that are not getting detected by the FTIR or NMR. The viscosity test performed
further supports that the crude biodiesel sample was successfully refined.
The heat of combustion results, Table 6, for crude biodiesel did not compare to the
B100 biodiesel. The potential cause for this difference is due to the impurities in the crude
biodiesel which is not allowing the fuel to produce more energy per gram. One possible
Crude Biodiesel 38.1 ± 0.2 KJ/g reason for the decrease in energy might

B100 Biodiesel 42.99 ± 0.03 KJ/g be from methanol, because the literature
value shows that it produces 22.689KJ/g.
Wet-Washed Biodiesel 41.9 ± 0.3 KJ/g
This is a lot lower than the B100 value so
Dry-Washed Biodiesel 42.7 ± 0.2 KJ/g
it might give a good indication that it is
Methanol30 22.689 KJ/g lowering the output of energy for the
Table 6. Measured Heat of Combustion Values and crude biodiesel. When the impurities were
Uncertainty Values for Crude, B100, Wet-washed,
Dry-washed Biodiesel, and Methanol. removed, the amount of energy produced
went up, for wet-washed and dry-washed biodiesel, showing what effect impurities have
on heat of combustion. The statistical test showed that the heat of combustion for wet-
washed and dry-washed biodiesel are statistically different. The dry-washed biodiesel
produces more energy per gram than wet-washed biodiesel. The heat of combustion test
 17

further supports that the crude biodiesel can be successfully refined and have comparable
results with B100 biodiesel.
 18

Conclusion
The main goal of this research was to show what effect impurities are having on
the properties of biodiesel and how the properties are changing when different refinement
methods are performed. The biodiesel was successful synthesized with a base-catalyzed
transesterification reaction using vegetable oil and the KOH/methanol solution. The FTIR
and NMR spectrums in Appendix A and B, respectively, show that the crude biodiesel was
identical to the B100 biodiesel with only the FTIR spectra showing methanol present in the
sample. Knowing the biodiesel was synthesized, the crude biodiesel was refined using the
wet-washing and dry-washing methods to rid the fuel of the leftover methanol from the
reaction. After refinement, both sample’s FTIR and NMR spectrums were taken and
produced the same results as both the B100 spectrum. This indicated that both refinement
methods produced viable biodiesel compared to the B100 sample.
Both refined biodiesel samples were tested against the ASTM values. For all the
characteristics done, both refined biodiesel samples fit within the range of values set by the
ASTM. This indicated, by the ASTM, that both refined samples could be used on the
commercial level. When looking at crude biodiesel, the impurities are shown to be
interfering with the properties. For the flashpoint and heat of combustion, it can be argued
that methanol was the impurity in the crude sample. When looking at the viscosity, its
shown that glycerol might be having an effect on the crude sample. Keep in mind, there
are other impurities, like water and catalyst, that might be present in the crude sample that
are not detected. For future work, samples of B100 biodiesel can be purposely
contaminated with each individual impurity and have them compared to one another. This
would be able to show which impurities are responsible for which properties. Another
study that could be done is to test other plant based oils, in order to find out which plant
based oil is the best in producing biodiesel. One last study could be on how many times the
water used in the wet-washing method can be used until it no longer refines the crude
biodiesel and the same goes for the magnesol beads in the dry-washing method. This study
brings the biodiesel research closer to refining biodiesel more cost efficiently to get it on
the market, to prevent the harmful emissions produced by fossil fuels from being released
into the atmosphere.
 19

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 21

Appendix A

FTIR Instructions:

The program OPUS is opened and the user name and password are plugged in.
1. Click the down arrow next to “Measurement” and click “Setup Measurement”
2. Click “Load” and select “Organic Lab”
a. Click “Accept and Exit”
3. Click “Measurement”
a. Type in sample description. Ex: Biodiesel MW
b. Click “Start Background Measurement”
c. Wait until bar hits 100%
4. Cover gold circle with sample
a. Click “Start Sample Measurement”
b. Wait until bar hits 100%
5. After Spectra is produced
a. Click “Baseline Correction”
b. Click “Peak Picking”
6. The spectra can now be saved, and the instrument is cleaned with acetone and wipe, for
addition runs.

FTIR Data:

O-CH2-C
Symmetry CH2 stretching and asymmetric
the asymmetry CH3 and CH2 axial
stretching stretching

C=O stretch

Figure 5. FTIR Spectra for Vegetable Oil.

 22

CH3
asymmetric
Symmetry CH2 stretching and bending O-CH3
the asymmetry CH3 and CH2 stretching
stretching C=O stretch

Figure 6. FTIR Spectra for B100 Biodiesel

OH stretch

CH3
O-CH3
asymmetric stretching
Symmetry CH2 stretching and bending
the asymmetry CH3 and CH2 C=O stretch
stretching

Figure 7. FTIR Spectra for Crude.

 23

CH3 O-CH3
Symmetry CH2 stretching and asymmetric
the asymmetry CH3 and CH2 bending stretching
C=O stretch
stretching

Figure 8. FTIR Spectra for Wet-Washed Biodiesel.

Symmetry CH2 stretching and CH3 O-CH3

the asymmetry CH3 and CH2 asymmetric
bending stretching
stretching C=O stretch

Figure 9. FTIR Spectra for Dry-Washed Biodiesel

 24

OH stretch

Figure 10. FTIR Spectra for Methanol

OH stretch

Figure 11. FTIR Spectra for Glycerol

 25

Appendix B

NMR Instructions for Bruker 300 MHz NMR:

The program NMR is opened on the computer. The following commands are
inputted into the computer. The command appears in quotation marks.
1. Click on “Spectra” and click on “1H”
2. Click on “Process” and click “Shim with AutoZero”
a. Wait until box turns grey
3. Type “zg” to collect data.
a. Wait until box turns grey
4. Open up NUTs program
5. Click on “Fourier Transfer”
6. Click on “Process” and click Zero Baseline
7. To set proper zero,
a. click and hold on desired peak
b. Type “O”
c. Type in 0 in box
d. Click “Okay”
8. Type “pp” for auto peak picking
a. To add/remove peaks
i. Type “dp”
ii. To add, click on peak
iii. To remove, hover mouse over peak and type “K”
 26

NMR Data:

Methyl Ester Shift

3.632

No
Methanol
Shift

Figure 12. NMR Spectra for B100 Biodiesel.

Methyl Ester Shift

3.654

No Methanol Shift

Figure 13. NMR Spectra for Crude Biodiesel

 27

Carbon Backbone

4.272, 4.227, 4.169

Figure 14. NMR Spectra for Vegetable Oil31

Methanol Shift
3.392

Methyl Ester Shift

3.655

Figure 15. NMR Spectra for B100 Biodiesel with Methanol

 28

Methyl Ester Shift

3.549

No
Methanol
Shift

Figure 16. NMR Spectra for B100 Biodiesel with Glycerol

Methyl Ester Shift

3.650

No
Methanol
Shift

Figure 17. NMR Spectra for Wet-Washed Biodiesel

 29

Methyl Ester Shift

3.648

No
Methanol
Shift

Figure 18. NMR Spectra for Dry-Washed Biodiesel