C H A P T E R

12
Biobased Fats (Lipids) and Oils from Biomass
as a Source of Bioenergy
Ciarán John Forde, Marie Meaney, John Bosco Carrigan, Clive Mills,
Susan Boland, Alan Hernon*
AER BIO, National Institute for Bioprocessing Research & Training (NIBRT), Blackrock, Co. Dublin, Ireland
*Corresponding author email: alan.hernon@aer-bio.com

O U T L I N E

Introduction 185 Hot Continuous Extraction (Soxhlet) 194

Countercurrent Extraction 194
Sources of Biolipids 186
Ultrasound Extraction (Sonication) 194
Plant-Derived Biolipids 186
Supercritical Fluid Extraction 194
Edible Lipids 186
Nonedible Lipids 187 Properties of Pure Plant Oil 195
Waste Edible oil 187 Degumming 195
Animal-Derived Biolipids 188 Alkaline Neutralization 195
Microalgae and Other Oleaginous Microorganisms- Winterization 195
Derived Biolipids 189 Bleaching 196
Transesterification 196
Supply and Projected/Purrent Volume 190
Properties of Biodiesel 196
Energy Balance 192
Biomass to Liquid Fuels (Bio-oil) 197
Processing of Biolipids and Properties of
Gasification 197
Biolipid-Derived Biofuels 193
Cleaning Process 197
Extraction 193
Synthesis 197
Steam Distillation 193
Maceration (Solvent Extraction) 193 Conclusion 198
Enzymatic Hydrolytic Maceration 193
References 198
Expression (Cold Pressing) 194

INTRODUCTION predators and for cooking. The earliest lamps recorded

Biolipids have been an important source of energy
since prehistoric times. While the term “biofuel” is
now often synonymously used with “biodiesel”, the first
biofuels used were wood or other plant materials, which
were burnt to provide heat, light, protection from
were made using plant material that was soaked with
animal fat, such as lard. Later lamps, which used oils,
were introduced in the eighteenth century, with early
lamp fuels being oils from fish, whale and a variety of
nut and other plant sources. Whale oil was much sought
after for a lamp fuel as it produced a cleaner flame with

Bioenergy Research: Advances and Applications 185

http://dx.doi.org/10.1016/B978-0-444-59561-4.00012-7 Copyright Ó 2014 Elsevier B.V. All rights reserved.
186 12. BIOBASED FATS (LIPIDS) AND OILS FROM BIOMASS AS A SOURCE OF BIOENERGY
less odor and smoke. Another source of light was
candles, which were made from tallow and other oils
rendered from animal waste. These fuels are known as
primary biofuels, fuels that are used without any signif-
icant processing in contrast to secondary biofuels where
significant processing is required before the raw prod-
ucts can be used as fuels. As they were discovered,
coal, gas and petroleum products (kerosene in partic-
ular) slowly replaced tallow and other animal-based
fuels. Similarly, the use of biolipids as transport fuel is
not novel; in fact, in 1900 when Rudolf Diesel showcased
his internal combustion engine at The Exposition
Universelle in Paris it was fuelled by peanut oil (Stauffer
and Byron, 2007). However, advancements in the use of
petroleum as fuel at the turn of the century resulted in
the selection of this abundant, cheap and efficient hydro-
carbon as the fuel of choice for transport. It was not until
the oil crisis of 1973 when oil became expensive and the
security of supply became paramount that biolipids
were investigated again; however, this interest was short
lived as the supply of crude oil from the Organization of
Arab Petroleum Exporting counties was restored in
1974. Now over 100 years after Diesel’s invention we
are almost completely dependent on this finite, expen-
sive and polluting hydrocarbon (petroleum) as a trans-
port fuel. Consequently, the use of petroleum-based
products has resulted in a significant number of envi-
ronmental issues including global warming via the
greenhouse gas (GHG) effect. Also, in an era when it is
generally accepted that we have reached peak oil
production and it is projected that the demand for trans-
port fuel will increase globally by 39% in the next
10 years interest in the use of biolipids as fuel has
reached new heights. Recent years have seen significant
research, investment and advances in sustainable en-
ergy technologies such as solar, wind, geothermal, tidal
and hydroelectrical. It should be noted, however, that
these energy sources, along with nuclear power, relate
to the generation of electricity. Currently electricity
only accounts for about 33% of the world energy market,
whereas liquid fuels account for the remaining 67% of
global energy consumption. These figures, along with
the finite nature of crude oil stocks, illustrate the need
to drastically increase the production of sustainable
liquid fuels (Schenk et al., 2008). Alternative liquid fuel
sources are continually being sought (Bereczky, 2012;
Singh and Singh, 2010) and while the obvious solution
is to revert to the use of vegetable oil used in 1900, there
are several problems with that approach. Most notably is
the need to use arable land to feed the world’s exponen-
tially growing population. Land use for the production
of liquid biofuels has become a hotly debated topic since
2007 when a combination of poor harvests and alloca-
tion of vast quantities of land for the production of
biofuel (mostly corn ethanol) resulted in a spike in world
food prices (Tenenbaum, 2008). The ease in supply of
food to the world market in 2007/2008 acted at an indi-
cator to what will happen in the future as the world’s
population increases beyond 8 billion people and we
struggle to meet the nutritional needs of humankind. It
will simply be impossible to grow enough terrestrial
crops to meet the worlds nutritional and energy needs.
It is therefore necessary to explore the use of biolipids
from all sources including lipids from plant, animal
and microalgae sources. Recovering lipids from waste
products like recovered vegetable oil and beef tallow
will also have a role to play in meeting our insatiable
demand for energy. Therefore, it is important to
judicially select biolipids that require the minimum
land usage (maximizing ton of oil per hectare) and lipids
with good fuel properties, as discussed below. In addi-
tion, the energy consumed in growing and recovering
the biolipid is also an important consideration when
selecting a biomass for the production of biofuel.
SOURCES OF BIOLIPIDS
Biolipids can be derived from plant, animal, oleagi-
nous microorganisms and algal sources. The composi-
tion of biolipids derived from each of these sources
differs greatly and has varying degrees of suitability to
the biofuel production industry. The major lipids
produced from each of these sources are listed below
and the degree of suitability to the production of biofuel
production is discussed.
Plant-Derived Biolipids
In 2007, 95% of world biodiesel was produced via
edible plant oils, which were supplied by the agricul-
tural industry, with the vast majority supplied by
rapeseed oil, 84% (Food and Agriculture Organization,
2008). Overall, plant lipids are divided into three major
categories: edible, nonedible and waste vegetable oils
described below.
Edible Lipids
The main edible oils used for biofuel production are
rapeseed, palm and soy bean oils. Edible oils have the
disadvantage of competing directly with food produc-
tion. The use of edible oils for the production of biodiesel
competes directly with the use of land for the produc-
tion of food and without proper planning results in
reduced food production (Gui et al., 2008). However,
the productivity from edible oils is high in terms of oil
yield and the quality of the resulting biofuel. The oil
yield from palm is the highest of the commonly grown
edible oil crops at 5 tons per hectare while rapeseed
 SOURCES OF BIOLIPIDS
produces 1 ton per hectare and soy bean 0.52 tons per
hectare. A high lipid yield is vital for the economical
production of biofuel from these plants. Although the
productivity from palm oil is particularly high its use
as a biofuel is limited as it is the world’s most commonly
used edible lipid and thus competition for the oil
between the food and biofuel industry would result in
an increase in the price of this oil (Lam et al., 2009). In
terms of the suitability for biofuel, palm oil has a high
degree of saturation and thus is not the most suitable
for biofuel production with the resulting fuel having
poor cold flow properties. However, the cold flow prop-
erties of a lipid can be altered by the use of cold filtration
(Kerschbaum et al., 2008) or alternatively the use of alco-
hols such as ethanol, isopropanol or isobutanol, which
results in the production of fatty alkyl esters with lower
freezing points and therefore improved cold flow prop-
erties (Dunn, 2009). There are also some environmental
and ecological concerns surrounding palm oil produc-
tion, with the clearing of rain forests to make way for
palm plantations. The plantation costs of edible oil crops
are relatively low with the exception of palm oil, which
has a higher cost; however, this is offset by the high oil
yield from the crop. The overall estimated energy
balance of rapeseed and soybean is similar at 3.7 and
3.4, respectively, while palm oil is significantly higher
at 9.6 due to the high yields (Food and Agriculture
Organization, 2008). Currently rapeseed oil is the most
commonly used plant oil used in biodiesel production
because it makes an excellent biofuel with excellent
cold flow properties. The main disadvantage of using
rapeseed oil is the growth of rapes is difficult and unsus-
tainable as it must be part of a one in five rotation due to
the large quantity of nutrients required for the growth of
the organism and the buildup of pathogens and disease
in the environment targeting rapeseed if grown
annually.
Nonedible Lipids
Nonedible oils that may be used in biofuel production
include Jatropha, Pongamia, jojoba, linseed and cotton
seed oil. Nonedible oils are not suitable for human
consumption due to the presence of toxic compounds
in the oils, for example, curicin present in Jatropha oils
is a toxic lectin. Biofuels from nonedible lipids have
many advantages over the edible alternative including
the ability of these organisms to grow in harsh nutrient-
and moisture-limiting conditions and the reduction in
carbon emissions. Nonedible oils are generally more
cost-effective as they do not have applications in food
production and thus are lower value oils, containing
low sulfur concentrations and low aromatic compound
concentrations and the lipids produced are biodegrad-
able (No, 2011). A disadvantage of using nonedible oils
187
is the large amounts of free fatty acids (FFAs) that cannot
be converted into biodiesel using an alkaline catalyst
(Demirbas et al., 2011).
Jatropha is one of the most widely used nonedible oils
due to the high potential yield of 0.5e12 tons per hectare
per year; the yield is highly effected by the conditions in
which it is produced, and the ability of the organism to
grow in harsh environmental conditions of low water
availability and low nutrient content (Francis et al.,
2005). The oil produced by Jatropha has good cold flow
properties due to the composition of the oil. The Jatropha
plant is a small tree and produces seeds with high lipid
content. In addition to the drought resistance within the
plant it is also pest tolerant and unpalatable to animals
and grows rapidly with a lifetime of 30 years; each of
these factors makes it a suitable choice for the produc-
tion of biofuels. The ability of the plant to grow in harsh
conditions led to Jatropha being considered a revolution-
ary plant that could provide the solution for the produc-
tion of large volume of lipids without competing with
the food industry. However, when grown in marginal
lands studies revealed that the number of seeds pro-
duced by the plant was quite low and although the
tree is capable of growing in low nutrient conditions,
the lipid production is low (Pandey et al., 2012). There-
fore, the economic returns of Jatropha grown on marginal
lands is low; however, growing the crop in developing
areas with poor land may be a viable method of produc-
tion of oil on a small scale. The energy balance from the
crop is also low if only the seeds are used for the produc-
tion of biofuel; however, the value is increased if all
components, for example, the husks are also utilized
(Prueksakorn and Gheewala, 2006).
Waste Edible oil
Waste edible oil (WEO) is the waste product of cook-
ing or frying foods. The disposal of WEO is difficult and
thus the use of WEO as a biofuel would both alleviate
the problem of disposal in addition to providing a
renewable source of biodiesel. WEO has a high volume
of FFAs, 0.5e15% in comparison with the 0.5% content
of refined virgin vegetable oil, which cannot be con-
verted to biodiesel using an alkaline catalyst as the
FFAs undergo a saponification reaction with the catalyst
thus reducing efficiency and yield (Knothe et al., 2005).
The problem may be overcome by using a supercritical
methanol transesterification for the transesterification
process rather than an alkaline catalyst (Kusdiana and
Saka, 2004). The volume of WEOs available is quite
high with approximately 1 million tons produced in
Europe each year while 10 million tons are produced
annually in the United States (Gui et al., 2008). WEO is
available two to three times cheaper than virgin
vegetable oils (Phan and Phan, 2008) and the high
188 12. BIOBASED FATS (LIPIDS) AND OILS FROM BIOMASS AS A SOURCE OF BIOENERGY
volume of WEO available means it a viable method for
biodiesel production. WEO has a higher estimated
energy balance than rapeseed and soybean of 5.8;
however, the value is lower than that of palm oil at 9.5
(Food and Agriculture Organization, 2008).
Animal-Derived Biolipids
As outlined above many biological sources can be
used for the generation of biofuels (Demirbas et al.,
2011; Vasudevan et al., 2005); however, one source of
biomass for the production of biodiesel that is often
overlooked is the waste fat from animals (e.g.
(Ali et al., 2012; Duku et al., 2011; Feddern, 2011;
Panneerselvam et al., 2011; Wisniewski Jr et al., 2010)).
Generally three broad categories of waste animal fats
are describeddtallow and related raw fats from process-
ing industries, yellow grease from waste cooking oil
used to cook, for example, chicken, and brown grease
that is obtained from traps used to prevent waste fats
and oils being released into the environment. Animal
fats can be sourced as room temperature solids or
semisolids from a variety of animals and include tallow
and suet (cattle and mutton), lard (pigs), schmaltz
(poultry especially chicken and goose), duck, fish oil
and dairy products (milk, butter) (Jayasinghe and
Hawboldt, 2011; Kerihuel et al., 2005; Mrad et al., 2012;
Panneerselvam et al., 2011; Wisniewski Jr et al., 2010).
It is also possible to reclaim waste animal fats from
wastewater (Awad et al., 2012a). Many of the properties
of animal fats used put to specific uses have been known
for a long time (Andés, 1898; Shahidi and Zhong, 2005).
A significant percentage of waste animal fat can be con-
verted to biodiesel using similar techniques to those
used for plant oils, the main process being transesterifi-
cation, described later (Prosková et al., 2009). The
triglycerides in animal fats are saturated, compared to
unsaturated plant triglycerides, and this has some impli-
cations when used as biodiesel. In particular the cloud
point, the temperature at which the oils solidify, is
higher for animal fats. However, when used as additives
to other sources of diesel, for example, 5% or 20%
biodiesel (B5 or B20 blends), the high cloud point does
not affect the blend overall.
Production of biodiesel from waste animal fats has
been shown using a variety of methods including a
novel, integrated method in which fat from lamb meat
is continuously extracted by supercritical CO2 followed
by enzymatic production of biodiesel (Schenk et al.,
2008). Feedstocks containing high levels of FFAs require
an additional preproduction step to convert the FFAs
into esters, which can subsequently be converted into
biodiesel. Waste sources that contain high levels of
FFAs require a separate step (acid catalyzed pretreat-
ment) before the base catalyzed reactions can be used
to provide maximal yields of biodiesel (Canakci and
Van Gerpen, 2001; Knothe et al., 2005; Popescu and
Ionel, 2011). Multistep processes using waste restaurant
oil and animal (pig) fat containing high levels of FFAs
can achieve high yields of biodiesel of up to 80% by
volume on a small scale (Math et al., 2010). Other high
FFA content oils, including used cooking oils, rendered
animal fat and some inedible plant oils (Mathiyazhagan
et al., 2011) can be processed in a similar fashion (Bakir
and Fadhil, 2011).
The feasibility and sustainability of using waste
animal fats as feedstocks for biofuel production has
been the subject of many studies in many areas, for
example, general studies (Demirbas, 2009; Nigam and
Singh, 2011), Australia (Puri et al., 2012), Ghana (Duku
et al., 2011), the United States (Groschen, 2002), Brazil
(Aranda et al., 2009), Ireland (Thamsiriroj and Murphy,
2010) and Hungary (Lakó et al., 2008). In addition, the
use of animal fats from waste tissue may also have envi-
ronmental benefits, such as being considered as a waste
management process and as a fuel source that does not
compete with food resources (e.g. soybean), the food
versus fuel debate. Table 12.1 shows typical values
reported for triglycerides in several animal fats in com-
parison to values for soy, a commonly used plant-
derived feedstock. In all cases, waste animal fats contain
high levels of the fatty acids that are capable of being
converted to methyl esters by transesterification
reactions to produce usable biodiesel. From a sustain-
ability point of view an estimate of the total annual US
production of animal fats as compared to plant-
derived oils is shown in Table 12.2.
Vegetable oils tend to be produced for human con-
sumption, whereas animal fats form part of a wide
group of animal by-products that are rendered into
many products that may be used in part for human con-
sumption (e.g. production of gelatin). All animal by-
products, including fats, are coded and classified
(Alakangas et al., 2011) according to their intended use
and animal fats not intended for human consumption
are controlled in the European Union by Regulation
(EC) No 1069/2009 and related legislation. Similarly,
TABLE 12.1 Percentages of Fatty Acids in Animal Fats
Fatty Acid Beef Tallow Pork Lard Chicken Fat Whale Soy
Myristic 14:0 1.4e6.3 0.5e2.5 1 4e8 e
Palmitic 16:0 20e37 20e32 25 7e12 w10
Palmitoleic 16:1 0.7e8.8 1.7e5 8 7e18 e
Stearic 18:0 6e40 5e24 6 1e3 w5
Oleic 18:1 26e50 35e62 41 28e32 w20
Linoleic 18:2 0.5e5 3e16 18 1e2 w50

TABLE 12.2 Total Annual Production of US Fats and Oils
Vegetable Oil Production (billion pounds per year)
Canola
Corn
Cottonseed
Soybean
Sunflower
Total Vegetable Oil
Animal Fats (billion pounds per year)
Edible Tallow
Inedible Tallow
Lard & Grease
Yellow Grease
Poultry Fat
Total Animal Fat
Source: U.S. Department of Agriculture, 2010; U.S. Census Bureau, 2010.
the storage of animal fats for use as fuels also needs to be
addressed. The storage of raw animal fat under unsuit-
able conditions can lead to oxidation and other undesir-
able chemical and microbial processes that can affect the
quality of the final biodiesel product. The stability of the
final biodiesel:diesel blend can also be affected by long-
term storage under unsuitable conditions, and additives
such as antioxidants might be added to improve stability
(Geller et al., 2008; Jain and Sharma, 2010).
With the advent of Bovine spongiform encephalopathy
(BSE) and more specifically Transmissible spongiform
encephalopathies (TSE), there is a greater need to monitor
human health issues when using waste animal fats for the
production of biofuel, at all stages of the production pro-
cess. The rendering industry recognizes that safe product
(fats) can only be supplied if certain standards are adhered
to (Woodgate and Van Der Veen, 2004). The raw materials
could well have microbial contamination including path-
ogenic bacteria and possibly prion material (Baribeau
et al., 2005; Brown et al., 2007; Bruederle et al., 2008; Greene
et al., 2007). There is also concern that prions will survive
the rendering process itself (Bruederle et al., 2008). These
concerns have in part led to the publication of guidelines
for the safe handling and use of biodiesels (National
Renewable Energy Laboratory, 2009).
Many trials of waste animal fat biodiesel-powered
engines have been published (Darunde Dhiraj and
Deshmukh Mangesh, 2012; Kleinová et al., 2011;
Panneerselvam et al., 2011; Varuvel et al., 2012). One trial
using public transport buses (Proc, 2006) showed that
the biodiesel does not have any harmful effects on
the engines at B5 and B20 mixes and also shows
SOURCES OF BIOLIPIDS 189
environmental benefit by way of reduced exhaust
pollutants. However, there are other potential health
and environmental issues in using animal fats as a feed-
1.04 stock for biodiesel production (Greene et al., 2007) and
2.49 the production of safe biodiesel is in part dependent
on a safe feedstock (Woodgate and Van Der Veen,
0.617
2004). Finally, the processes involved (e.g. rendering,
19.61 cleanup, transesterification, etc.) in the production of
0.731 biodiesel will generate waste that also needs to be
assessed (Ellis, 2007).
24.49
Microalgae and Other Oleaginous
1.859 Microorganisms-Derived Biolipids
3.299
Microalgae are a heterogeneous group of organisms
1.63 consisting of both prokaryotes such as cyanobacteria
1.40 and eukaryotes such as diatoms (Bacillariophyta),
dinoflagelates (Dinophyta), green algae (Chlorophyta),
1.42
yellow-green algae (Xanthophyta), and red algae
9.61 (Rhodophyta) (Brennan and Owende, 2010; Hu et al.,
2008). Similarly, other oleaginous microorganisms are
defined as microorganisms with lipid content in excess
of 20%. The number of bacteria that produce lipids that
could be used for biodiesel production is very small. As
a result, bacteria are mainly used for special lipid produc-
tion such as Docosahexaenoic acid (DHA). Many yeasts
and fungi also produce high quantities of lipid. Yeasts
with high lipid content include Candida curvata (58%),
Cryptococcus albidus (65%), Lipomyces strakeyi (64%) and
Rhodotorula glutinous (72%). Oleaginous fungi include
Aspergillus oryzae (57%), Mortierella isabellina (86%), Humi-
cola lanuginose (75%) and Mortierella vinacea (66%) (Meng
et al., 2009). In terms of microalgae, species are generally
unicellular organisms but there are also a number of sim-
ple multicellular organisms that occur as colonial or fila-
mentous groups of cells. Microalgae are capable of
autotrophic, heterotrophic and mixotrophic growth.
Microalgae populate a wide variety of ecological niches
due to a wide range of tolerance for various growth con-
ditions such as availability of nutrients, salinity, pH and
temperature (Brennan and Owende, 2010; Gong and
Jiang, 2011; Schenk et al., 2008). Currently, microalgae
contribute very little biolipid to the overall bioenergy
market as full-scale commercialization has yet to be
realized. Despite this fact, microalgae remain the feed-
stock with the greatest potential for supplying future
demand for bioenergy in the form of liquid fuels. The
idea of using microalgae as a source of biolipids for bio-
fuel is not a new one, however. For example, the Aquatic
Species Program was launched in 1978 by what is now
known as the National Renewable Energy Laboratory
(NREL) with its main focus being, “the production of bio-
diesel from high lipid-content algae grown in ponds, uti-
lising waste CO2from coal fired power plants” (Sheehan
et al., 1998). Over 3000 microalgae strains were initially
190 12. BIOBASED FATS (LIPIDS) AND OILS FROM BIOMASS AS A SOURCE OF BIOENERGY
collected, 300 of which were eventually identified as oil
rich. When the program was officially closed in 1998
the conclusions were that no “fundamental engineering
and economic issues” were identified that would hamper
the feasibility of large-scale microalgae culture. The au-
thors noted, however, that total biomass and algal lipids
produced were still below “theoretical potential, and the
requirements for economic viability” (Sheehan et al.,
1998). The economic viability was, of course, based on a
time when oil prices in the United States were among
their all-time lowest at less than $20 per barrel (adjusted
for inflation). Today the average oil price is approxi-
mately $100 per barrel and this, along with increased
pressure to reduce GHG emissions as well as significant
technical advances, has made microalgae-derived bio-
fuels even more relevant to meet current bioenergy
demands.
SUPPLY AND PROJECTED/PURRENT
VOLUME
Growing microalgae for biolipid production usually
involves a lag phase of growth followed by a stationary
phase induced by some sort of “stress” This “stress”,
often nitrogen depletion, induces a switch in the meta-
bolism of the microalgae, which encourages the produc-
tion of storage lipids in the form of triacylglycerides
(TAGs) rather than cell division (Meng et al., 2009;
Widjaja et al., 2009). Currently microalgae can be grown
at industrial scale autotrophically in open raceway
ponds (Sapphire Energy, 2013) or closed photobioreactor
(PBR) systems (Solix BioSystems, 2013). In addition,
many microalgae species have the ability to grow
heterotrophically, in closed fermenters, given a suitable
carbon source (Solazyme Inc., 2013). Open culture sys-
tems, such as race way ponds, are significantly lower
cost in terms of capital expenditure. They require greater
land area than closed systems and are more prone to
contamination by invasive species. Water loss due to
evaporation can also be a significant problem when
compared to closed systems (Chisti, 2007; Pulz, 2001;
Sheehan et al., 1998). Closed systems, on the other
hand, such as PBRs or fermenters are by their nature
closed and thus less likely to be contaminated. Nutrient
concentration can be more easily controlled and water
loss through evaporation is negligible. However, some
have argued that loss of cooling water, used to control
temperature, negates any savings made from using a
closed culture system. The tighter control over culture
conditions facilitated by a closed culture system, along
with more sterile cultures, results in PBRs producing
much greater levels of microalgae biomass, when
compared to raceway ponds. However, the increased
production capability must be offset against the much
larger capital cost involved in commissioning and main-
taining a closed culture system (Carvalho et al., 2006;
Pulz, 2001; Ugwu et al., 2008). Hybrid systems have
also been proposed whereby a closed system is used
for the log phase production of biomass and the nutrient
depleted lag phase is allowed to occur in large raceway
ponds. It is hoped that the relatively concentrated inoc-
ulation of the raceway ponds will not allow any invasive
species to become established (Greenwell et al., 2010;
Huntley and Redalje, 2007; Rodolfi et al., 2008).
Microalgae present significant potential as a
source of biolipids for bioenergy over more traditional
sources of biolipids such as palm, soya or Jatropha for a
number of reasons. Firstly, the oil content of microalgae
as a percentage of the dry weight, shown in Table 12.3,
is generally in the range of 20e70%, although levels
above 40% are rarely observed (Borowitzka, 1988).
Similarly, the potential yield of biolipids and derived
biodiesel from microalgae per area far outweighs that
of any current oilseed crop. For example, one of the
best available studies of large-scale algae cultivation
produced 0.1 g/l day or 20e23 g dry weight/m2 day.
A conservative lipid content of 30% could therefore
yield 24,000 l biodiesel/ha year (Moheimani and
Borowitzka, 2006; Schenk et al., 2008). This compares
extremely favorably with both Jatropha (1892 l
biodiesel/ha year) and oil palm (5950 l biodiesel/ha
year) (Schenk et al., 2008).
The high potential yield of biodiesel from microalgae-
derived biolipids is due to a number of factors including
the growth rate of microalgae (Scott et al., 2010) all year
round production capability (Schenk et al., 2008) and the
higher photon conversion efficiency compared to terres-
trial plants (Melis, 2009). Unlike algae-derived biofuels,
first-generation biofuels directly competed with food
crops for arable land sparking the “Food vs Fuel” debate
(Gui et al., 2008). Although second-generation fuel crops
such as Jatropha can grow on marginal land (Francis
et al., 2005), microalgae are capable of growing on
nonarable land ensuring competition for land with
food crops is significantly reduced. Similarly, in terms
of other resource demands, 1 kg of algae biomass
requires 1.83 kg of CO2 to grow (Chisti, 2007) and
much research has investigated the potential of indus-
trial flue gases as a source of this CO2 (Bilanovic et al.,
2009). This possibility of both sequestering excess CO2
from flue gases that would otherwise be released into
the atmosphere, while also increasing the growth rate
of microalgae to be used for bioenergy, offers both
environmental and economic advantages (Pires et al.,
2012; Yun et al., 1997). More recently, the apparent
“peak phosphorus” problem has been identified
whereby phosphorus will become a limiting resource
in agriculture. As a result, the potential industrial scale
culture of microalgae, which requires a phosphorus
 SUPPLY AND PROJECTED/PURRENT VOLUME 191
TABLE 12.3 Lipid Content and Biomass Productivity of Biofuel Relevant Algae Species

Lipid Content Biomass Productivity

Algae Species (% Dry Weight) (g/l day) References

Botryococcus braunii 25e75 (Chisti, 2007)

Chlorella protothecoides 15e58 1.214 (Gong and Jiang, 2011)
Chlorella emersonii 63 (Gouveia and Oliveira, 2009)

Chlorella minutissima 57 (Gouveia and Oliveira, 2009)

Chlorella protothecoides 55 (Gouveia and Oliveira, 2009)
Chlorella sorokiana 22 (Gouveia and Oliveira, 2009)
Chlorella sorokiniana 19e22 0.045 (Gong and Jiang, 2011)
Chlorella sp. 28e32 (Chisti, 2007)
Chlorella vulgaris 56 (Gouveia and Oliveira, 2009)

Chlorococcum sp. 19 0.054 (Gong and Jiang, 2011)

Crypthecodinium cohnii 20 (Chisti, 2007)
Cylindrotheca sp. 16e37 (Chisti, 2007)
Dunaliella bioculata 8 (Gouveia and Oliveira, 2009)
Dunaliella primolecta 23 (Chisti, 2007)
Dunaliella salina 6e25 (Gong and Jiang, 2011; Gouveia and
Oliveira, 2009)
Ellipsoidion sp. 27 0.047 (Gong and Jiang, 2011)

Isochrysis sp. 25e33 (Chisti, 2007)

Monallanthus salina 20 (Chisti, 2007)
Nannochloris sp. 20e35 0.038e0.061 (Chisti, 2007)
Nannochloropsis oculata 22e30 0.084e0.142 (Gong and Jiang, 2011)
Nannochloropsis sp. 31e68 (Chisti, 2007; Gong and Jiang, 2011)
Neochloris oleoabundans 29e65 0.090e0.134 (Chisti, 2007; Gong and Jiang, 2011;
Gouveia and Oliveira, 2009)
Nitzschia sp. 45e47 (Chisti, 2007)
Pavlova lutheri 36 0.05 (Gong and Jiang, 2011)

Pavlova salina 31 0.049 (Gong and Jiang, 2011)

Phaeodactylum tricornutum 18e57 0.045 (Chisti, 2007; Gong and Jiang, 2011)
Scenedesmus dimorphus 16e40 (Gouveia and Oliveira, 2009)
Scenedesmus obliquus 35e55 (Gouveia and Oliveira, 2009)
Scenedesmus sp. 20e21 0.041e0.054 (Gong and Jiang, 2011)
Schizochytrium sp. 50e77 (Chisti, 2007)

Spirulina maxima 4e9 (Gouveia and Oliveira, 2009)

Tetraselmis sueica 15e23 (Chisti, 2007)

and nitrogen source for growth, would also be affected Commercial harvesting of algae blooms from waste-
(Cordell et al., 2009). Both phosphorus and nitrogen water has already been demonstrated in New Zealand
are available in plentiful supply within waste water (Aquaflow, 2013) and the use of wastewater streams as
streams (Sawayama et al., 1995; Yun et al., 1997). a nutrient source in large-scale cultivation of microalgae
192 12. BIOBASED FATS (LIPIDS) AND OILS FROM BIOMASS AS A SOURCE OF BIOENERGY
has been well studied and implemented. Similarly, in
terms of water usage, microalgae cultivation, particu-
larly in closed cultivation systems, demonstrates signif-
icant water savings when compared to traditional
biofuel crops. Many microalgae species are also capable
of growing in brackish water most notably Dunaliella
salina (Weldy and Huesemann, 2007).
ENERGY BALANCE
Any credible source of bioenergy should not only be
economically viable but also environmentally sustain-
able. The economic and environmental impacts of any
source of bioenergy, including biolipids from microal-
gae, will usually be measured in terms of energy return
on energy investment (EROI) and/or GHG emissions.
These economic and environmental impacts of biofuels
and microalgae biofuels in general have been hotly
debated in recent years. A number of life cycle analyses
(LCAs) have been undertaken with seemingly conflict-
ing results (Benemann et al., 2012; Liu et al., 2011;
Resurreccion et al., 2012; Sun et al., 2011). Similar
disparities arose in the case of second-generation bio-
fuels such as corn ethanol before the introduction of
the Energy and Resources Group (ERG) Bioenergy
Meta-Model (Farrell et al., 2006). The results of reported
LCA analyses are hindered by the lack of fully inte-
grated commercial-scale microalgae to bioenergy sys-
tems from which to obtain accurate measurements.
Estimates are based on projections from laboratory-
and pilot-scale tests, as well as some commercial data.
Despite these facts an overall meta-analysis concluded
that algae-based biodiesel would result in energy con-
sumption and GHG emissions on par with terrestrial al-
ternatives (Liu et al., 2011). In this study the authors
consider a microalga-based bioenergy system whereby
CO2 and nitrogen for microalgae cultivation are recycled
from waste streams and the microalgae coproducts are
used for further bioenergy production in the form of
methane. This concept of an integrated “biorefinery”
has been proposed previously (Borowitzka, 1995, 1999;
Chisti, 2007; Martı́n and Grossmann, 2012). As alluded
above, the “biorefinery” concept envisages the main in-
puts into the cultivation process such as carbon, nitro-
gen and phosphorus being supplied through various
waste streams. Similarly, the microalgae product result-
ing from cultivation could be fully “refined” into a num-
ber of outputs including biolipids for bioenergy,
biolipids for nutraceutical applications, proteins for an-
imal feeds, sugars for bioethanol production, etc. At pre-
sent, where fully commercial scale cultivation of
microalgae and conversion to fuel alone is still not
economically feasible, the “biorefinery” concept appears
to offer the best short to medium term path to scale-up.
In addition to the potential economic and environ-
mental advantages of using microalgae-derived bio-
lipids, the properties of the resulting biodiesel product
are also worth considering. As detailed later in this
chapter, biodiesel is produced by transesterification of
the biolipids from an appropriate feedstock. Much like
the plant- and animal-based biolipids discussed previ-
ously, the profile of the microalgae-derived biolipids
that undergo transesterification will ultimately deter-
mine the quality of the biodiesel product. This profile
will include the level of polyunsaturated fatty acids
(PUFAs), the level of FFAs and the level of TAGs.
Although the lipid profile of microalgae varies among
species and even among the same species under
different conditions of growth, approximately 80% of
the lipid content of microalgae, in general, will be
made up of storage lipids in the form of TAGs. TAGs
are made up of three fatty acid chains, usually with a
chain length of C14 to C22 for microalgae-derived bio-
lipids, joined to glycerol through three ester bonds (Scott
et al., 2010). These TAGs can be easily transesterified in
the presence of methanol, as described later in the chap-
ter, to fatty acid methyl esters (FAMEs), which make up
biodiesel. The presence of FFAs, however, results in the
formation of soaps during transesterification in the pres-
ence of a base catalyst such as NaOH. This increases the
downstream processing required to produce a finished
biodiesel product. Similarly, the presence of PUFAs in
biolipids derived from some microalgae species can
cause tar formation resulting from fatty acid chains
cross-linking (Burton et al., 2009). A high PUFA content
could also mean that a biodiesel product would not pass
European standards for biodiesel (EN14214), which de-
mand the content of FAMEs with four or more double
bonds to be below 1% mol (Knothe et al., 2005). Other
properties that have been considered with regard to
other feedstocks mentioned in this chapter include the
cloud point, the cetane number and the oxidation stabil-
ity of the biodiesel fuel. It has been suggested that bio-
diesel from microalgae oils may face significant
performance problems regarding cold flow and oxida-
tive stability in particular (Knothe, 2011); however,
exceptions to this observation may apply to some micro-
algae such as Trichosporon capitatum. Also, in a recent
study, biodiesel derived from the microalgae Chaetoceros
gracilis was found to generate similar torque and power
to soy-derived biodiesel. In terms of emissions, the
C. gracilis-derived biodiesel also produced less CO,
NOx and hydrocarbons than petroleum diesel (Wahlen
et al., 2012).
It is clear that the potential for algae to supply a sus-
tainable source of biolipid for transportation fuel and
other forms of bioenergy is not in doubt. However, there
remain technical, economic and environmental chal-
lenges to be overcome. In a recent report by the National
 PROCESSING OF BIOLIPIDS AND PROPERTIES OF BIOLIPID-DERIVED BIOFUELS
Research Council in the United States entitled, “Sustain-
able development of algal biofuels” a number of sustain-
ability concerns were highlighted. These included EROI;
GHG emissions and resource usage such as land, water,
nitrogen, phosphorus, and carbon dioxide (National
Research Council, 2012). None of these concerns,
however, were considered a “definitive barrier to sus-
tainable development of algal biofuels”. This is because
a number of strategies have already been implemented
to tackle these challenges. As mentioned previously
the use of wastewater streams can drastically reduce
resource usage and GHG emissions as well as greatly in-
crease EROI. Current projects, at industrial scale, such as
Sapphire Energy’s “Green Crude Farm” (Sapphire
Energy, 2013) aim to have a capacity of 1 million gallons
per year of finished biofuel product. It is predicted that
this will result in a 60e70% reduction in GHG emissions
compared to traditional fossil crude oil, which, if
achieved, will make the potential of microalgae-
derived biofuel a very definite reality.
PROCESSING OF BIOLIPIDS AND
PROPERTIES OF BIOLIPID-DERIVED
BIOFUELS
Independent of the biomass source, biolipids can be
used in various ways as a source of bioenergy. There
are a number of basic steps involved in processing bio-
lipids to biofuel. These can include some or all of lipid
extraction, degumming, neutralization, winterization,
bleaching and transesterification. The sources of
biomass and how they are produced have been
described previously in this chapter and the first pro-
cessing step will usually involve efficient extraction of
the biolipid from the biomass. Following extraction,
some biolipids can be used in their pure form as pure
plant oils (PPOs). Other biolipids are further processed,
usually into biodiesel. Here the extraction step is
followed by purification and stabilization of the biolipid
and the conversion to biodiesel. The various steps
involved in processing biolipids are described below,
beginning with extraction, along with the fuel properties
of both PPO and biodiesel.
Extraction
Extraction is a process consisting of the separation of
a specific substance from a complex matrix. In the
context of extraction lipids from biomass, the purpose
is to use standardized extraction procedures to isolate
the biomolecules of interest, i.e. lipids, concurrent to
rejection of the remaining inert biomass. This is most
commonly achieved by using a selective solvent known
as menstruum (Handa, 2008), or by solventless physical
193
extraction means. The resultant lipid may be ready for
use in the form of fluid extracts, it may be further pro-
cessed into a variety of biofuel and nutraceutical prod-
ucts, or it may be fractionated to isolate individual
chemical entities or a combination of the above as
proposed by the “biorefinery” concept discussed previ-
ously. The most common biolipid extraction procedures
are summarized below.
Steam Distillation
Steam distillation is a process that is commonly
applied to the extraction of essential oils (Gutierrez
et al., 2009). Plant material is placed into a still where
pressurized steam penetrates the plant material causing
internal lipid vacuoles to rupture. Upon exposure to the
surrounding environment, the lipid evaporates to form a
mixture of easily separable vapors (essential oil and
water). The vapors condense and the distillate (sepa-
rated into two immiscible layers) is collected in a gradu-
ated tube connected to a condenser. The aqueous phase
is recirculated into the flask, while the volatile oil is
collected separately. The main disadvantage associated
with steam distillation is that thermolabile components
risk being degraded (Sarker et al., 2005). A combination
of solvents and steam distillation is often used to
improve the final product of a biodiesel production
process.
Maceration (Solvent Extraction)
Maceration is used for creating extracts and resins in
a simple yet well-established procedure. Whole or
coarsely powdered biomass is placed in intimate contact
with a suitable extractant in a closed vessel. The mixture
is allowed to stand at room temperature for a defined
period of time, typically at least 3 days, with frequent
agitation (using mechanical shakers or mixers) to ensure
homogeneity (Sarker et al., 2005). The organic phase is
separated from the solids by either filtration, decanta-
tion or in some cases centrifugation and the remaining
solid material is pressed to ensure efficient solvent
recovery. The recovered liquid phases are combined
and clarified for further processing. This process can
be repeated several times to achieve maximum lipid
recovery. The main disadvantage associated with macer-
ation is that the process can be quite onerous, potentially
taking from a few days up to several weeks (Takahashi
et al., 2001).
Enzymatic Hydrolytic Maceration
Certain plant materials require enzymatic maceration
prior to lipid release as their volatile components
are glycosidically bound. Enzymes can be either
194 12. BIOBASED FATS (LIPIDS) AND OILS FROM BIOMASS AS A SOURCE OF BIOENERGY
endogenous or exogenous to the biomass. For example,
methyl salicylate (wintergreen oil) is an organic ester
that is naturally produced by many species of plants.
The plant leaves contain the precursor gaultherin and
the enzyme primeverosidase; when the leaves are
macerated in warm water, the endogenous enzyme
acts on the gaultherin and liberates free methyl salicylate
and primeverose (Handa, 2008). In the case of the exog-
enous addition of enzymes, recent advances in the field
of algal lipids have demonstrated the addition of com-
plex mixtures of enzymes to selectively degrade cell
walls in a cascade of hydrolytic reactions. Released
lipids are isolated and collected for further processing
(Liang et al., 2012).
Expression (Cold Pressing)
Expression or cold pressing is commonly used in the
production of essential and food oils. The term expres-
sion refers to any physical process in which the essential
oil glands in the biomass are crushed or broken to
release the oil. The resulting oilewater emulsion is typi-
cally separated by centrifugation. Traditionally, cold
pressing was conducted by hand; however, for large-
scale commercialization, this is impractical. Thus, with
the advancement of industrialization, a number of
machines have been designed to achieve the same
results on commercial scale. It is important to note that
oils extracted using this method have a relatively short
shelf life (Martı́nez et al., 2008).
Hot Continuous Extraction (Soxhlet)
In this method, finely ground biomass is placed in a
porous bag or “thimble” made of strong cellulose, which
is placed in the extraction chamber of a Soxhlet appa-
ratus. The menstruum is heated, and the condensed
extractant drips into the thimble containing the biomass,
ensuring intimate continuous contact with the biomass.
When the level of liquid in the extraction chamber
reaches overflow, the liquid contents siphon into the
heating chamber. This process is continuous and is
carried out until complete extraction is achieved
(Morrison and Coventry, 2006). The advantage of this
method is that large amounts of lipid can be extracted
with a much smaller quantity of solvent.
Countercurrent Extraction
Counter-current extraction is a process whereby wet
raw material is pulverized using toothed disc disinte-
grators to produce slurry in a semicontinuous stream.
As the pulverization of the biomass is in aqueous media,
the heat generated during comminution is counterbal-
anced by the slurry water, preserving thermolabile
compounds. The slurry stream is moved in one direction
within the cylindrical extractor where it comes
into discreet contact with a suitable menstruum
(Vishwakarma, 2010). Complete extraction is possible
when the quantities of solvent and material and their
respective flow rates are optimized. The quantity of
solvent required is generally minimal and as the process
is most often conducted at room temperature, the threat
to thermodegradation of volatile compounds is negated
(Handa, 2008).
Ultrasound Extraction (Sonication)
The use of sonication is an emerging technology
that is gaining widespread industrial acceptance due
to recent advances in the scalability of the technology
(Awad et al., 2012b; Dolatowski et al., 2007). In the
context of lipid extraction from biomass, ultrasound
technology is used to increase the permeability of
biomass cell walls by generating cavitation events.
These events are created by the use of high frequencies
(20e2000 kHz) to generate a microbubble in solution;
the intensity of the waves leads to the eventual
collapse of the bubble generating extreme localized
pressure and temperature events in close proximity
to the biomass. These cavitation events assist in the
rupturing of the cell walls to release the intercellular
constituents into the surrounding environment. Once
the biomolecules of interest are released from the
biomass they can be recovered using conventional
techniques. One disadvantage of using ultrasonics in
the occurrence of sonolysis, i.e. the occasional but
deleterious effect that when high power (typically
greater than 20 kHz) is applied in aqueous media it
can lead to the formation of free radicals and hydrogen
peroxide. These are generated at the interfacial double
layer established during cavitations, which subse-
quently diffuse into solution (O’Donnell et al., 2010).
Supercritical Fluid Extraction
Another technology in the extraction space is super-
critical fluid extraction (SFE) whereby a solvent is
subjected to temperature and pressure conditions to
adjust the properties to those intermediate to a gas
and liquid in a dedicated reactor setup. This in turn
effects the solubilization of solutes in a matrix
(Wenclawiak, 1992). The main supercritical solvent
employed is carbon dioxide. Carbon dioxide (critical
conditions: T ¼ 30.9  C and P ¼ 73.8 bar) is cheap, envi-
ronmentally friendly and has generally recognized as
safe status from the US Food and Drug Administra-
tion. Supercritical CO2 (SC-CO2) is also attractive
because of its high diffusivity combined with its
easily tunable solvent strength (Herrero et al., 2010).
 PROPERTIES OF PURE PLANT OIL
However, due to its chemical nature, it possesses
several polarity limitations. As mentioned previously,
solvent polarity is particularly important when
extracting polar solutes and when strong matrix inter-
actions are present. To augment the process, organic
solvents are commonly added to the carbon dioxide
extracting fluid to alleviate the polarity limitations
(Handa, 2008). CO2 is gaseous at room temperature
and pressure, which makes recovery very simple and
provides solvent-free products, i.e. once the liquid
depressurizes, the CO2 returns to a gaseous state,
and only the extracted products remain. SFE using
CO2 can be operated at low temperatures, which
allows the extraction and integrity preservation of
thermolabile compounds (Mendiola et al., 2007).
PROPERTIES OF PURE PLANT OIL
Following extraction from biomass, biolipids can be
used as pure oil (generally plant) or can be converted
to biodiesel by a process known as transesterification,
described later. However, the use of PPO as a fuel re-
quires the modification of diesel engines unlike bio-
diesel, which, particularly when blended with
petroleum diesel, can be used in unmodified diesel
engines. These engine modifications are needed as
PPO is more than 10 times as viscous as biodiesel. As
a result, it has a tendency to gum up in cold weather,
which can be somewhat overcome by blending with
traditional fossil diesel. Nevertheless, it has some
advantages: with a flash point of over 300  C, storage
and transport are simplified. According to the VwVwS
(Verwaltungsvorschrift wassergefährdende Stoffe),
which is the national German regulation on water
hazard classification, PPO is not designated as even a
hazard to water given that it is biodegradable. In an
unmodified engine, poor atomization of the fuel will
lead to coking of the injectors and accumulation of
soot deposits. Modification is designed to preheat fuel
or involves installation of a two-tank system. In the
latter, the engine is started with diesel and only changes
to PPO when the operating temperature has been
reached. It must switch back to diesel before being
turned off, to flush out the remainder of the PPO in
order to ready the engine for the next operation. Other
options exist, such as the specialist engine developed
by Ludwig Elsbett in the 1970s. The fuel emissions of
PPO are also much lower in sulfur emissions when
compared to the fossil equivalent. For a detailed over-
view see (Russo et al., 2012). After extraction, if the
biolipid is not to be used as PPO, or other pure oil, it
needs to be further processed into a more useable bio-
fuel, usually biodiesel. Here the biolipid goes through
a series of processing steps beginning with degumming.
195
Degumming
Following extraction and regardless of the process
described above, the end product will generally be a rather
impure biolipid that contains undesirable contents such as
FFAs, tocopherols, waxes and possibly phosphatides. The
latter, if not removed before storage, will produce a thick
gum over time. Gums are formed when the biolipid
absorbs water, which causes some of the phosphatides
(such as phosphocholine) to become hydrated and thereby
lipid insoluble. Accordingly, hydrating the gums and
removing the hydrated gums from the oil before storage
can prevent the formation of a gum deposit. This treat-
ment is called water degumming and involves the addi-
tion of water at 60e90  C before the phase is separated.
An optimum temperature is sought, as it must not be so
high as to increase the solubility of phosphatides in oil.
A temperature that is too low will increase the viscosity,
making phase separation more difficult. It is never applied
to fruit oils like olive oil and palm oil, since these oils have
already had considerable water contact during their pro-
duction. The removal of nonhydratable phosphatides
(such as phosphatidic acid) requires the addition of an
acid, usually citric or phosphoric, which will form a sludge
that can be easily removed (Dijkstra and Van Opstal, 1989).
This addition of acid is proportional to the amount of
phosphorous already contained in the sample. In addi-
tion, this acid also reduces any iron salts and decreases
chlorophyll contamination. Enzymatic degumming fo-
cuses on the use of lipases, which convert nonhydratable
lipids to more hydratable forms. Although the process
has been tried at a larger scale for 20 years, it has not
made the advancement toward widespread use (Dijkstra,
2010; Yang et al., 2008).
Alkaline Neutralization
As mentioned previously, the presence of FFAs in the
biolipid is detrimental to oil quality and function,
including biodiesel production. Removal typically in-
volves the reaction of these FFAs with an alkaline solu-
tion. In the edible oil industry, usually only caustic
soda is used for this reaction, but potassium hydroxide
is also used by some producers. The acidity of the FFA
comes from the Hþ of the carboxyl group. This Hþ of
the functional group of the stearic acid reacts with the
OH group of the caustic soda (NaOH) to produce
soap and water. In addition to the removal of FFAs, other
undesirable nonglyceride materials are also removed in
this fashion such as phenol, oxidized fatty compounds,
heavy metals and phospholipids.
Winterization
Most biolipids do not need dewaxing, as they contain
little or no waxes. Only biolipids of higher melting
196 12. BIOBASED FATS (LIPIDS) AND OILS FROM BIOMASS AS A SOURCE OF BIOENERGY
temperatures, such as sunflower oil and rice bran oil,
give a hazy appearance during winter season due to
precipitation of dissolved waxes. Hence, they require
being dewaxed. This is carried out by chilling the oil
to 10e15  C, followed by filtration of precipitated solids.
The oil thus treated has a sparkling appearance, even in
winter temperatures.
Bleaching
Oil bleaching, which is performed in order to prepare
a sufficiently light-colored product of enhanced appear-
ance and improved stability, is usually achieved by
treating the crude or refined oil with powdered
absorbent. These absorbents usually contain a calcium
montmorillonite (fuller’s earth) or natural hydrated
aluminum silicate (bentonite). Adsorption of color
bodies, trace metals and oxidation products, as well as
residual soaps and phospholipids remaining after
washing neutralized oils takes place, if possible. Acid-
activated clays are the major adsorbent used, although
active carbons and synthetic silicas are also applied
industrially with more specific goals. Thus, active
carbons are used specifically to eliminate polycyclic
aromatic hydrocarbons from some oils, especially fish
oils and pomace oils, while synthetic silicas are quite
efficient in adsorbing secondary oxidation products,
phospholipids and soaps (León-Camacho et al., 2003).
There are a number of good sources of material with
more detailed descriptions of each process found online
at the Lipid Library (Hardwood and Weselake, 2013), in
“Proceedings of the World Conference on Oilseed
Technology and Utilization” (Applewhite, 1993) and
finally in, Edible Oil Processing (Hamm and Hamilton,
2000).
Transesterification
Despite being energetically favorable, the direct use
of plant or other biolipids in fuel engines is problematic
as described earlier. Briefly, due to high viscosity (over
10 times higher than diesel fuel) and low volatility,
they do not burn efficiently and can form deposits in
the fuel injector of diesel engines. Furthermore, acrolein
(a highly toxic substance) is formed through thermal
decomposition of glycerol. Different ways have been
considered to reduce the high viscosity of plant and
other biolipids, but the principal method is to engage
in chemical transesterification to produce biodiesel,
which could be used in the common diesel engine
with minor modification.
As mentioned previously, biolipids consist primarily
of triglycerides, which are three hydrocarbon chains
connected by glycerol. The bonds are hydrolyzed to
allow the formation of FFAs, which are mixed and
reacted with methanol or ethanol to form methyl (or
ethyl) fatty acid esters. The use of methanol (methanol-
ysis) is widespread and considered advantageous, as it
is cheaper than ethanol (although in Brazil, ethanol 90
is plentiful) and has less azeotrophic qualities (Encinar
et al., 2007). The same reaction using ethanol is more
complicated as it requires a water-free alcohol, as well
as a biolipid with low water content, in order to obtain
good glycerol separation. Methanolysis can happen by
heating 80e90% methanol with a small amount of cata-
lyst. The received biodiesel after methanolysis is FAME
and with ethanol to form fatty acid ethyl ester. The use of
ethanolysis reaction using bioethanol has been
discussed as being possibly more environmentally
favorable as it would involve the use of a nonfossil
fuel. Apart from this, ethanol is less toxic and slightly
increases the cetane number of the biofuel. Although
transesterification can proceed in the absence of
catalysts, the reaction proceeds much too slowly to be
economically viable and thus typically requires an acidic
or alkaline catalysis. Among the most commonly used
alkaline catalysts in the biodiesel industry are potassium
hydroxide (KOH) and sodium hydroxide (NaOH)
flakes, which are inexpensive, easy to handle and can
be transported and stored easily. For this reason, they
are preferred by smaller producers. Alkyl oxide solu-
tions of sodium methoxide (NaOCH3) or potassium
methoxide (KOCH3) in methanol, which are now
commercially available, are the preferred catalysts for
large continuous-flow production processes.
In the transesterification process, the effective species
of catalysis is the methoxide radicals (CH3O) and the
activity of a catalyst depends upon the amount of meth-
oxide radicals (Komers et al., 2001a,b). For sodium or
potassium hydroxide, the methoxide ion is prepared in
situ by reacting methanol with hydroxide, a reaction
that will also produce water that remains in the system.
Hydrolysis of triglycerides and alkyl esters may occur
due to the presence of this water, which further leads
to the formation of FFAs and thus to a soap. Saponifica-
tion may also occur if a strong base, e.g. NaOH or KOH,
is present in the system by reacting with esters and
triglycerides directly. All these problems can be avoided
completely if sodium and potassium methoxide
solutions, which can be prepared water-free, are applied
(Singh et al., 2006).
PROPERTIES OF BIODIESEL
Untreated biodiesel blend stocks, generated by trans-
esterification, generally exhibit poor oxidation stability,
which can result in long-term storage problems.
Biodiesel has many similar fuel economies to fossil
diesel. Although it has about 10% less energy content
 BIOMASS TO LIQUID FUELS (BIO-OIL)
per volume, its cetane number and lubricating effect are
higher, which is advantageous (Rutz and Janssen, 2007).
The higher oxygen content leads to better combustion
and fewer pollutants, particularly sulfur oxides. Bio-
diesel is produced in a pure form (100% biodiesel blend
stock, referred to as “B100” or “neat biodiesel”) and is
typically blended with petroleum-based diesel fuel.
Such biodiesel blends are designated as BXX, where
XX represents the percentage by volume of pure
biodiesel contained in the blend (e.g. “B5” or “B20”).
According to a “Technical Statement on the Use of
Biodiesel in Compression Ignition Engines” released in
2009 by the Truck and Engine Manufactures Associa-
tion, neat biodiesel and higher percentage biodiesel
blends can cause a variety of engine performance
problems. These include fuel filter plugging, injector
coking, piston ring sticking and breaking, elastomer
seal swelling and hardening/cracking, and severe
engine lubricant degradation and dilution. The report
goes on to state that when converting from petroleum-
based diesel to a biodiesel blend, residual fuel system
deposits may accumulate in fuel filters due to the high
solvency of the fuel. Thus, more frequent filter service
may be required until the fuel system deposits are stabi-
lized. More information on biofuel handling can be
found in "Biodiesel Handling and Use Guide: Fourth edition
(Revised)" published by NREL (National Renewable
Energy Laboratory, 2009).
BIOMASS TO LIQUID FUELS (BIO-OIL)
While the focus of this chapter has been on biolipids it
is important to note that any biomass can be converted to
“bio-oil” via a high-temperature process known as pyrol-
Ò
ysis. This “bio-oil” also known as Synfuel or Sunfuel is
currently only produced on a small scale and it very
much belongs to the second-generation biofuels, as it is
a way of generating fuel from a range of biomass
including straw, wood or other materials high in lignin,
which are difficult to convert to bioethanol. The potential
for mass production remains enormous. The production
of this biomass to liquid or BtL fuel can vary in
complexity and can vary depending on the individual
needs, but it essentially comprises the following steps.
Gasification
Gasification is a form of incomplete combustion in
which a fuel is burnt in an oxygen-deficient atmosphere.
An energy-rich gas, consisting principally of methane,
CO and hydrogen, is formed but heat release is mini-
mized. Thus an energy-rich fuel (biomass) is converted
into an energy-rich gas. There are differing processes
for gasification. For example, a description of the
197
Ò
Carbo-V process first developed by Chloren Industries
but now owned by Linde Engineering GmbH was out-
lined in the Biofuel Technology Handbook (Rutz and
Janssen, 2007). This involves low-temperature gasifica-
tion, where low-temperature pyrolysis with air or oxy-
gen at 400e500  C allows the continuous production of
a gas containing both tars (volatile component) and
char (carbon solids). This is followed by a high-
temperature gasification, where the gas is further
oxidized (again hypostoichiometrically) in a combustion
chamber. The third part involves blowing the pulverized
char into the hot gasification medium. Pulverized char
and gasification medium react endothermically in the
gasification reactor and are converted into a raw synthe-
sis gas. Other gasification processes can be found, such
as the recently developed BioliqÒ , which was formed
by Lurgi AG (Frankfurt Germany) with Karlsruhe Insti-
tute of Technology (Karlsruhe Germeny).
Cleaning Process
After gasification, it is usual to have many impurities
and thus cleaning remains one of the most important
and most technical challenges. Remaining tars tend to
be refractory and difficult to remove by thermal or phys-
ical processes. Generally, the impurities in biosyngas pro-
duced from the gasifier can be grouped into three types:
(1) organic impurities, such as tars, benzene, toluene, and
xylenes; (2) inorganic impurities, such as O2, NH3, HCN,
H2S, Carbonyl sulfide (COS), and HCl; and (3) other im-
purities, such as soot and dust. Both thermal cracking,
which involves the addition of steam and oxygen at
200e1000  C, and catalytic cracking at lower tempera-
tures is possible, as is low-temperature scrubbing with
an oil-based medium may all encompass the process. A
multicontaminant syngas treatment process created by
Southern Research Institute, Birmingham, Alabama,
USA, uses a candle filter, which can be catalytic, closely
coupled with the gasifier. A variety of sorbents is injected
into the gasifier or between the gasifier and filter to
remove various contaminants (e.g. alkali metals, sulfur
species, and halides) both by reaction in the gas phase
and on the filter cake. Catalysts may be incorporated
into the candle filter or the filter may be coated with a
catalyst to crack tar and ammonia depending on the oper-
ating temperature of the candle filter. An outline of the
process can be seen in Figure 12.1.
Synthesis
Two methods are available for this production step,
but the Fischer-Tropsch (FT) synthesis is the most
widely known. It was developed at the Kaiser-Wilhelm
Institute for Research on Coal (Mühlheim/Ruhr) in
1925. In Germany, coal to liquid fuels have been
198 12. BIOBASED FATS (LIPIDS) AND OILS FROM BIOMASS AS A SOURCE OF BIOENERGY
produced with the help of FT synthesis since 1938.
During the process, CO and H2, with the aid of a
catalyst, will form hydrocarbons. A variety of catalysts
exist, but the most common are usually transition metals
such as cobalt. In the case of biomass, however, an iron
catalyst is often favored (Hu et al., 2012).
The other process is the methanol-to-gasolineÒ
method, in which the syngas is first transformed into
methanol as an intermediate state. In a following step,
fuels can be obtained from this compound. Finally, after
separating the produced liquid hydrocarbons into
heavy, medium and light fractions, these hydrocarbons
are refined and blended to achieve the desired fuel
properties.
CONCLUSION
The search for a sustainable supply of fuel that does
not contribute to global warming has consumed envi-
ronmental scientists for decades. While it is unlikely
there is a “silver bullet” solution to the pending energy
crisis the use of biolipids has enormous potential to
meet a large proportion of the global transport fuel
requirements. Similarly, no Single lipid source is pro-
duced in sufficient quantities to impact on the world’s
fuel supplies; therefore, a combination of all biolipids
outlined above will be required if biolipids are to be a
realistic alternative to petroleum-based fuels. While
plant-derived biolipids currently dominate the liquid
bioenergy markets, microalgae remain the most prom-
ising source of biolipids in the future. The limited land
FIGURE 12.1 Biobased fats (lipids)
and oils from biomass as a source of
bioenergy. Integration of a catalytic filter
into a gasifier for combined particle
separation and tar removal from
biomass gasification gas. Source: Cour-
tesy of Southern Research Institute, Bir-
mingham, Alabama, USA. (For color
version of this figure, the reader is
referred to the online version of this
book.)
usage requirement and efficient carbon fixing capabil-
ities of microalgae make them the ideal choice as a
source of biolipids; however, there are a number of
stumbling blocks to be overcome before algal biofuels
are a commercial reality. These include the challenge of
growing algae at industrial scale to meet the increasing
demand for liquid transport fuel, the energy input
involved in harvesting and dewatering algae and finally
the cost and environmental impact of efficiently extract-
ing biolipids from algae. These challenges are far from
insurmountable, however, and each challenge is being
tackled by numerous academic institutions and increas-
ingly, by large, multinational energy, food and industrial
chemical companies. This concerted effort with regard
to algae biofuels, coupled with the more established
plant- and animal-based biofuel industries can supply
a significant portion of the world’s energy needs in the
future.
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