Waste Management 30 (2010) 18811888

Contents lists available at ScienceDirect

Waste Management
journal homepage: www.elsevier.com/locate/wasman

Analyzing alternative bio-waste feedstocks for potential biodiesel production using

time domain (TD)-NMR
Rebecca M. Willson a,b, Zeev Wiesman a,*, Asher Brenner b
a

Phyto-Lipid Biotechnology Laboratory, Department of Biotechnology Engineering, The Institutes for Applied Research, Ben-Gurion University of the Negev, P.O. Box 653,
Beer-Sheva 84105, Israel
b
Unit of Environmental Engineering, Ben-Gurion University of the Negev, P.O. Box 653, Beer-Sheva 84105, Israel

a r t i c l e

i n f o

Article history:
Received 21 December 2009
Accepted 9 March 2010
Available online 27 March 2010

a b s t r a c t
Production of biodiesel is currently limited due to lack of economically benecial feedstocks. Suitability
of municipal wastewater sludge and olive mill waste as feedstocks for biodiesel production was evaluated. The various bio-waste sources were analyzed for their oil content and fatty acid composition using
conventional analyses complemented with time domain (TD)-NMR analysis. TD-NMR, a rapid nondestructive method newly applied in this eld, yielded good correlations with conventional methods.
Overall biodiesel yields obtained by TD-NMR analysis were 7.05% and 9.18% (dry wt) for olive mill pomace and liquid wastes, and 11.92%, 7.07%, and 4.65% (dry wt) for primary, secondary, and anaerobically
stabilized sludge, respectively. Fatty acid analysis indicated fundamental suitability of these agro-industrial waste resources for biodiesel production. Evaluation of bio-waste materials by TD-NMR revealed the
potential of this tool to identify waste-oil sources cost effectively and quickly, supporting expansion of a
sustainable biodiesel industry in Israel and other regions.
2010 Elsevier Ltd. All rights reserved.

1. Introduction
Increased global energy consumption along with instability in
the price of conventional energy sources has led to a renewed
interest in alternative energy sources. Extensive use of fossil fuels,
the most popular energy source for transportation, has had an
cumulative negative impact on the environment (Houghton et al.,
2001), resulting in a desire for clean energy to replace fossil fuels.
Another environmental concern is the increasing production of
wastes by human societies and industries around the world. Large
quantities of highly polluted wastes accumulate, threatening soil
and freshwater resources due to toxic substances and high nutrient
and organic content (including oils and fats). Moreover, disposal of
wastes is restricted by strict regulations (Fytili and Zabaniotou,
2008), making reduction and clean up of these wastes important
to all societies.
In this context, biomass has a strong potential as a natural and
renewable source of energy. One of the promising fuels that can be
produced from biomass is biodiesel, a renewable and carbon neutral alternative to petroleum fuels. Biodiesel is biodegradable and
nontoxic, and has low emission proles, reducing its negative impact on the environment (Dorado et al., 2003). This alternative fuel
is currently produced from oil crops, waste cooking oil, and animal
fat (Demibras, 2005). However, production of biodiesel from these
* Corresponding author. Tel./fax: +972 8 647 7184.
E-mail address: Wiesman@bgu.ac.il (Z. Wiesman).
0956-053X/$ - see front matter 2010 Elsevier Ltd. All rights reserved.
doi:10.1016/j.wasman.2010.03.008

resources cannot realistically satisfy even a small fraction of the

existing demand for transport fuels (Chisti, 2007). Lack of agricultural lands for growing biodiesel feedstocks limits the expansion of
this alternative. World oil and food prices have risen dramatically
over the past few years and this has impacted the economic possibilities of biodiesel. Moreover, the relatively high production costs
of biodiesel compared to fossil-based diesel currently restrict its
commercialization on a large scale (Ma and Hanna, 1999). In order
to expand the biodiesel industry, research has been carried out to
investigate alternative oil feedstocks, such as oil yielding crops
grown in arid climate and marginal lands (Chapagain et al.,
2009), oil yielding algae farms (Chisti, 2007), and reutilization of
wastes (Dufreche et al., 2007). The worlds energy industry would
benet from a low-cost, high-energy, and renewable feedstock
supply for efcient biodiesel production.
A promising and environmentally benecial feedstock for biodiesel is bio-waste material, such as food production residues
and sewage sludge (Hepbasli et al., 2003; Mondala et al., 2009).
Organic wastes, generated in large volumes around the world,
contain substantial amounts of oil and fats that pose great
challenges to treatment and disposal strategies. Reutilization of
bio-wastes as alternative energy holds the possibility of reducing
their impact on the environment along with the potential to
expand the currently limited biodiesel industry.
Olive mill waste treatment and disposal is one of the most serious environmental problems in the Mediterranean countries,
which account for over 97% of the worlds olive oil production

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R.M. Willson et al. / Waste Management 30 (2010) 18811888

(Kiritsakis, 1998). Large quantities of highly polluting olive mill

wastes, in terms of land, water reservoirs and air quality, currently
lack a generally adopted viable treatment and disposal strategy
(Roig et al., 2006). Over the past 20 years the Israeli olive oil industry has undergone signicant growth. However, the downfall of the
newly emerging industry is the dramatic increase in olive mill
wastes produced, creating a new environmental problem for Israel
waste management (www.sviva.gov.il/olivemill wastewater). A recent approach suggested the use of olive mill wastes for the production of alternative energy (Hepbasli et al., 2003). The large
and signicant quantities generated solid and liquid waste and
the high oil content indicate the elds nancial potential. As was
previously stated (Dorado et al., 2003), for exhaust emission proles of waste cooking olive oil derived biodiesel, hydrocarbon fuels
made from olive residue have negligible sulphur content, and thus
a reduced impact on the environment. Olive oil residue is a renewable source of energy that has been converted into biofuel materials using thermochemical technologies (Roig et al., 2006). Cookingwaste olive oil, exhibiting a similar composition to olive mill waste
oils (Kiritsakis, 1998), has recently been proven to be a suitable
feedstock for biodiesel (Dorado et al., 2003).
Biological treatment of industrial and municipal wastewater
has been gaining credibility around the world due to improved levels of water purication and effective stabilization of organic matter and pathogenic content (Fytili and Zabaniotou, 2008). The main
waste by-product of this process is sludge, generated in vast quantities on a daily basis around the world. Moreover, the amount of
sewage sludge is expected to increase due to urbanization and
industrialization (Mondala et al., 2009). The handling of sewage
sludge is currently one of the most signicant challenges in wastewater management (Fytili and Zabaniotou, 2008). In Israel, 47 municipal activated-sludge treatment facilities treat over 75% of the
wastewater generated in the country (www.sviva.gov.il/waste
water), resulting in an estimated 500,000 tons/year of stabilized
sludge (semi-solid, approximately 7580% solids). The costs of
sewage sludge treatment and disposal are increasing and have
been evaluated to be in the order of 50% of the total wastewater
treatment costs (Boocock et al., 1992). Alternative new technologies may offer the greatest potential for cost reduction through improved energy recovery. Research has indicated that lipids
contained in sewage sludge may be a potential feedstock for biofuel production (Boocock et al., 1992). More recently, biodiesel
has been successfully produced directly from sewage sludge lipids
(Dufreche et al., 2007). These researchers tested various methods
of extracting lipids from secondary sludge, achieving an optimal
biodiesel yield of 4.41% using a solvent mixture extraction. Mondala et al. (2009) tested in situ biodiesel production from sludge under various conditions, achieving biodiesel yields of 14.5% and 2.5%
for primary and secondary sludge, respectively.
Further testing of the suitability of food residue waste, sewage sludge, and other wastes is needed in order to promote biodiesel production from these untapped benecial lipidfeedstocks to answer the growing demand for biodiesel. Technologies for wide-scale screening of potential biodiesel feedstock
are needed to identify suitable candidates. Currently the most
common oil analysis technologies are quantitative solvent
extraction methods such as Soxhlet combined with qualitative
gas chromatograph (GC) and gas chromatography mass spectroscopy (GCMS) fatty acid analysis (Casado et al., 1998). Conventional oil analysis methods are typically tedious, time
consuming, and environmentally unfriendly due to their consumption of large quantities of organic solvents. Advanced technologies such as 1H nuclear magnetic resonance (NMR) may aid
the search for alternative biodiesel feedstocks by making oil
testing of various materials more reliable, efcient, and environmentally friendly (Gao et al., 2008).

Time domain (TD)-NMR is a fast, non-destructive, and accurate

method widely used in the oilseed and food industry for the evaluation of oil and fat content (Nordon et al., 2001; Todt et al., 2006).
Compared to high resolution NMR requiring highly homogeneous
magnetic elds, low resolution NMR experiments are obtained by
permanent magnet technology, which signicantly reduces overall
system and running costs (Todt et al., 2006). TD-NMR applications
involve complex macromolecule structures that behave differently
when exposed to magnetic elds, as they relax at different rates.
ISO 10565 (1998) is an ofcial TD-NMR method suited for oil and
water content determination in oilseeds. In the standard method,
application of a spin-echo NMR pulse sequence separates the lipid
hydrogen nuclei signal from other hydrogen nuclei signals, which
enables determination of the samples lipid content after appropriate calibration (Gao et al., 2008). Application of this technology has
been broadened to many industries, such as petroleum, polymer,
and agricultural (Viereck et al., 2008). Recently, Gao et al. (2008)
successfully applied this method for measuring lipid content in algae. However, the use of TD-NMR for evaluation of lipids in wastes
has received little attention. Therefore, the scope of this research
was to assess the feasibility of using TD-NMR for oil content evaluation in bio-waste materials.
As part of the search for alternative feedstocks for wide-scale
biodiesel production, we have tested olive mill wastes and municipal wastewater sludge as potential oil feedstocks for biodiesel production, using the advanced technology of TD-NMR. Quantitative
conventional oil analysis by Soxhlet extractor was complemented
with TD-NMR to enable wide-scale evaluation of the wastes oil potential. Qualitative fatty acid analysis by GC and GCMS was conducted to determine suitability of the waste oils for biodiesel
production.
2. Methods
2.1. Chemicals
Methanol, acetone, n-hexane, heptane, sulphuric acid, and sodium chloride of analytical grade were purchased from Bio-lab,
Jerusalem, Israel. Chemicals were used as received.
2.2. Sample preparation
Olive mill pomace (OMP) and wastewater (OMW) samples were
collected from 10 olive oil mills across Israel operating a threephase oil extraction process (consisted on the following fractions:
oil; water waste and pomace), constituting a representative 10%
of olive oil mills in Israel. Primary, secondary, and anaerobically
stabilized sludge samples were collected from a municipal wastewater treatment plant in the city of Beer-Sheva, Israel. Secondary
sludge samples, produced by an activated-sludge process, were
collected after partial thickening by gravity belt thickener. Stabilized sludge, produced by anaerobic digestion of primary and secondary sludge, was sampled after belt lter press dewatering.
Sampling of municipal wastewater sludge was conducted over a
one-year period to account for seasonal variations in sludge
composition.
After collection, the water content of the waste samples was removed to avoid interference during oil analyses. Sample drying
was conducted by an industrial air forced oven operating at 70 C
for about 45 days until full dryness. Following the drying procedure, samples were mechanically ground to ne particles using a
Hsiangtai electric grinder (Taipei Hsien, Taiwan) in order to maximize the waste materials surface area. Samples were stored in airtight containers under refrigeration. Moisture content was
determined on a weight basis.

R.M. Willson et al. / Waste Management 30 (2010) 18811888

2.3. Organic solvent extraction

The total oil content of olive mill waste and municipal wastewater sludge was determined gravimetrically by Soxhlet extractor
method. Dried, nely ground waste samples of 50 g were washed
by 500 ml of organic solvents: n-hexane for olive mill wastes,
and a solvent mixture (20% methanol, 20% acetone and 60% hexane) for municipal wastewater sludge, for efcient lipid extraction
from sewage sludge (Dufreche et al., 2007). Soxhlet oil extraction
was carried out in a Soxhlet apparatus for 24 h. The weight of oil
was measured gravimetrically after removing the extracting solvent by a rotary evaporator (4000WB, Laborata, Germany). The
yield of extracted material was then determined and expressed
as the percentage of extractable oil from dry solids on a weight basis. Each experiment was conducted in three repetitions to the
samples yield average oil content.
2.4. TD-NMR analysis
All NMR measurements were performed on a Maran bench-top
pulsed NMR analyzer (Resonance Instruments, Witney, UK),
equipped with a permanent magnet and 18 mm probe head, operating at 23.4 MHz. Prior to NMR analysis, pre-dried samples
(<10 wt% moisture content) were equilibrated at 40 C for 30 min.
The standard TD-NMR oil analysis method (ISO 10565, 1998)
was used. The spin echo pulse sequence used 90 and 180 pulse
widths of 6 and 12 ls, respectively, and pulse spacing (s) of
3.5 ms. Each run consisted of 16 scans with a recycle delay of 1 s,
and acquisition of a single data point at t = 7 ms.
The TD-NMR instrument was separately calibrated for each of
the two bio-wastes. TD-NMR sampling of olive mill wastes (OMP
and OMW) and municipal wastewater sludge (primary, secondary,
and stabilized sludge) comprise individual waste streams, sampled
in equal quantities for each testing. Each set consisted of 20 samples; 10 were used for constructing the calibration curves (training
set), and the remaining 10 samples were used for validating the
model (testing set). For each measurement, the NMR acquired signal was weight standardized and cross-correlated to the reference
oil content obtained by the Soxhlet method to construct a linear calibration curve. This calibration was stored in the NMR computer
program enabling the evaluation of unknown waste sample oil content using NMR analysis alone. Goodness of the t (R2) was tested
by comparing the calculated versus measured oil content in a linear
regression t. After training and testing of each waste, TD-NMR
analysis was used to characterize the oil content of individual waste
stream. All TD-NMR screenings were conducted in three replicates.
NMR sampling values represent average values of the screenings.
2.5. Transesterication
Conversion of olive mill waste and municipal wastewater
sludge oils to fatty acid methyl esters (FAMEs), i.e., biodiesel, was
carried out through chemical transesterication reaction by two
catalysis methods. Alkaline-catalysis transesterication was used
for olive mill waste samples according to AOCS ofcial method
Ce 2-66 (AOCS, 1997) slightly modied by Kaufman and Wiesman
(2007). Due to higher free fatty acid value than olive oil mill waste,
conversion of municipal wastewater sludge lipids to FAMEs was
carried out through acid-catalysis transesterication using a modied version of Christies method (Christie, 2003) proposed by
Dufreche et al. (2007).
2.6. Fatty acid analysis
Qualitative analysis of olive mill waste and municipal wastewater sludge oil fraction was conducted by GC and GCMS fatty acid

1883

analyses. Prior to analysis, fatty acid methyl esters of the oils were
prepared by the transesterication procedures detailed in Section
2.5. The waste oils were analyzed for their fatty acid composition
using a Varian star 3400 gas chromatograph with a ame-ionization detector (Palo Alto, CA, USA). Column dimensions were
15 m  0.32 mm, with phase thickness of 0.25 lm.
GCMS was used to identify individual components of municipal wastewater sludge lipids. FAMEs were analyzed using a 6890N
model gas chromatograph (Agilent Technologies) equipped with a
30 m  0.25 mm i.d., 0.25 lm column. Fatty acids were identied
both by their retention time and by mass spectra obtained with a
5973 Network Mass Selective Detector (Agilent) operating at
70 eV according to the GCMS library.
Biodiesel (FAME) quantication of the two wastes was conducted by the internal standard method using heptadecanoate acid
(C17:0) as an internal standard. The European standard method EN
14103 was used for olive mill waste samples (EN 14214, 2003). A
modied version of EN 14103 proposed by Schober et al. (2006),
which accounts for the natural content of C17:0 in the sample,
was used for municipal wastewater sludge samples.
2.7. Statistical analysis
Statistical analysis was carried out on a minimum of three replicates for all experiments.
Ten samples of OMP and OMW, and 10 samples of each sludge
type (primary, secondary, and stabilized) were analyzed by conventional and TD-NMR oil analysis. Individual waste sources were
characterized for their moisture content, oil yield, and overall biodiesel yield by average values of all single samples tested, presented as mean standard error.
3. Results and discussion
3.1. TD-NMR analysis
TD-NMR was tested as a tool for qualitative characterization of
olive mill waste and municipal wastewater sludge oil content.
TD-NMR training procedure yielded the two waste representative
calibration curves, presented in Fig. 1a. High correlations of
R2 = 0.983 and R2 = 0.979 were achieved between TD-NMR screening values and Soxhlet oil content for olive mill waste and municipal wastewater sludge, respectively. The relationship between
the oil content determined by TD-NMR and Soxhlet methods
using the testing set of samples is illustrated in Fig. 1b. A linear
regression model was obtained for both wastes with correlation
coefcients of R2 = 0.995 for olive mill wastes and R2 = 0.993 for
municipal wastewater sludge. The high correlation coefcients
obtained suggest that TD-NMR is as successful for oil quantication of the two wastes as the conventional Soxhlet oil analysis,
commonly used to analyze oil content in various materials.
Characterization of the two wastes oil content by TD-NMR is presented in Table 1. Average oil yields by TD-NMR were 12.25% and
16.97% (dry wt) for olive mill pomace and wastewater, respectively, and 23.35%, 18.65%, and 12.61% (dry wt) for primary,
secondary, and stabilized sludge, respectively. Results obtained
by TD-NMR are compatible with oil yields obtained by the conventional Soxhlet method for all waste streams tested (Table 1).
These results demonstrate the suitability of TD-NMR for oil quantication of bio-waste materials such as olive mill waste and municipal wastewater sludge. TD-NMR is a fast, easy to use, and
non-destructive method (Todt et al., 2006), and may be benecial
when implemented for the evaluation of alternative biodiesel
feedstocks. This technology has the potential for wide-scale
screening of waste materials to broaden the characterization of

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R.M. Willson et al. / Waste Management 30 (2010) 18811888

Fig. 1. (a) TD-NMR calibration of olive mill pomace and wastewater (R2 = 0.983),
and of primary, secondary, and stabilized municipal wastewater sludge (R2 = 0.979),
illustrating a linear correlation between conventional Soxhlet oil content and TDNMR screening values. (b) Oil content determined by conventional Soxhlet analysis
in relation to TD-NMR analysis. Correlation coefcients were of R2 = 0.995 for olive
mill waste and R2 = 0.996 for municipal wastewater sludge.

untapped environmentally benecial oil sources for future biodiesel production.

3.2. Oil and biodiesel yields
Quantitative characterization of olive mill waste and municipal
wastewater sludge oil and biodiesel potential was conducted by
the conventional Soxhlet method and the advanced technology of

TD-NMR. Results obtained by both methods were compared, further testing the novel application of TD-NMR for oil analysis of
waste materials. Average moisture content, oil yields, saponiable
oil, and overall biodiesel yields of olive mill waste and municipal
wastewater sludge are presented in Table 1.
Olive mill waste generated average oil yields by Soxhlet extraction of 11.39% and 17.41% (dry wt) for olive mill pomace and
wastewater, respectively. Oil yields obtained by TD-NMR are comparable to these results, differing by less than 1% (Table 1a). Taking into account moisture content, these values change to 5.57% for
OMP and 1.16% for OMW on a wet weight basis, exhibiting slightly
greater oil content than values reported in literature of 4% and 1%
for OMP and OMW, respectively (Hepbasli et al., 2003). Comparing
the oil yields of the two olive mill wastes on a wet weight basis implies a greater potential of OMP for biodiesel production due to signicantly lower moisture content (<50%). OMPs lower moisture
content will result in lower drying costs, reinforced by the fact that
the oil extraction process from this waste to produce low grade olive oil is well known (Roig et al., 2006). These factors make OMP a
benecial feedstock for future biodiesel production. Overall biodiesel yields, accounting for both oil and transesterication yields
(saponiable matter), were evaluated based on oil yields obtained
by Soxhlet and TD-NMR methods (Table 1). Olive mill waste overall
biodiesel yields by Soxhlet extraction method were 6.57% and
9.40% (dry wt) for OMP and OMW, respectively. TD-NMR analysis
obtained overall biodiesel yields of 7.05% and 9.18% (dry wt) for
OMP and OMW, respectively. Strong similarity between the results
obtained by both methods (<0.5% difference) show the suitability
of TD-NMR for quantication of oil and biodiesel potential in olive
mill wastes (Table 1a). Overall biodiesel yields obtained by both
methods show that olive mill wastes can be reutilized as a benecial alternative feedstock for biodiesel production.
Municipal wastewater sludge oil yields evaluated by Soxhlet
and TD-NMR methods are presented in Table 1b. Average oil yields
by Soxhlet extraction were 23.11%, 17.83%, and 13.24% (dry wt) for
primary, secondary, and stabilized sludge, respectively. Oil yields
obtained by TD-NMR for all three sludge types were comparable
to these results differing by less than 1% as presented in Table
1b. Overall biodiesel yields using the Soxhlet extraction method
were 11.88%, 6.76%, and 4.88% (dry wt) for primary, secondary,
and stabilized sludge, respectively. TD-NMR analysis obtained
overall biodiesel yields of 11.92%, 7.07%, and 4.65% (dry wt) for primary, secondary, and stabilized sludge, respectively. Comparing
the overall biodiesel yields obtained by the two methods shows a
strong similarity between the results (<0.5% difference), indicating
the fundamental suitability of TD-NMR for oil and biodiesel

Table 1
Moisture, oil, saponiable matter, and biodiesel yields of olive mill waste and municipal wastewater sludge by Soxhlet and TD-NMR analysis.

a
b
c
d

Waste material

Moisturea (%)

Oil analysis method

Oil yielda (%)

Saponiable
matterb (%)

Overall biodiesel yielda (%)

(a) Olive mill waste

Olive mill pomace

49.25 2.40
93.33 1.41

11.39 2.35
12.25 1.42
17.41 2.01
16.97 1.74

57.53

Olive mill wastewater

Soxhlet
TD-NMR
Soxhlet
TD-NMR

6.57 1.56
7.05 0.85
9.40 1.49
9.18 0.53

(b) Municipal wastewater sludge

Primary sludge

96.38 1.32
94.46 1.72

Stabilized sludged

80.75 1.54

23.11 3.72
23.35 2.67
17.83 2.48
18.65 1.98
13.24 1.48
12.61 1.23

51.04

Secondary sludgec

Soxhlet
TD-NMR
Soxhlet
TD-NMR
Soxhlet
TD-NMR

Average moisture, oil, and overall biodiesel yields are presented as mean standard error.
Percentage of transesteried oil determined by internal standard method (Schober et al., 2006).
Moisture content following thickening by a gravity belt thickener.
Moisture content following dewatering by a belt lter press.

54.1

37.89
36.88

11.88 1.41
11.92 0.78
6.76 1.67
7.07 0.98
4.88 1.04
4.65 0.46

R.M. Willson et al. / Waste Management 30 (2010) 18811888

quantication of municipal wastewater sludges (Table 1b). It

should be emphasis that sludge composition typically varies
depending on the origin of the wastewater and the specic treatment applied, rationalizing the different oil and biodiesel yields
obtained by other studies (Fytili and Zabaniotou, 2008).
Among the three sludge types tested, primary sludge achieved
the greatest overall biodiesel yield, with an average of 11.88%
and 11.92% by Soxhlet and TD-NMR methods, respectively (Table
1b). Mondala et al. (2009) also achieved greater biodiesel yields
for primary sludge in comparison to secondary sludge, though
their results were obtained by in situ biodiesel production, a process where biodiesel is directly produced from dry sludge. Primary
sludge consists of relatively fresh organic matter originating from
the raw wastewater prior to biological processing (Fytili and Zabaniotou, 2008). Therefore, it is logical that primary sludge contains
the largest quantity of oil and most intact saponiable material.
Comparing overall biodiesel yields of the three sludge types shows
primary sludge as the most benecial feedstock for biodiesel production. Moreover, using primary sludge for biodiesel production
may reduce energy consumption of further biological processes
for both wastewater and sludge, and decrease the total amount
of sludge residual.
3.3. Fatty acid analysis
Qualitative analysis of olive mill waste and municipal wastewater sludge oils was conducted to determine the suitability of these
oils for biodiesel production. GC and GCMS fatty acid analysis
yielded characteristic fatty acid proles of the wastes. The oils
fatty acid composition is essential when testing the suitability of
potential feedstock for biodiesel production, since it directly inuences the produced fuel properties (Knothe, 2005).
A characteristic GCMS fatty acid prole representing an average of primary, secondary, and stabilized sludge proles is presented in Fig. 2. Mass spectra proles of the main fatty acid
components identied in the three types of sludge are presented
as linked boxes above the fatty acid peaks. GCMS analysis enabled

1885

the identication of 14 fatty acids in municipal wastewater sludge

ranging from C11:0 to C24:0 (Fig. 2). The main FAME components,
which consist of over 90% of the fatty acid composition, range between myristic (C14:0) and linoleic (C18:2) fatty acids. GCMS identication of municipal wastewater sludge fatty acid composition
was dominated by: 16:0 (MW = 256.42 and main MS nger prints:
58, 83, 194, 235m/z); 16:1 (MW = 254.41 and main MS ngerprints: 74. 143, 227, 270m/z); 18:0 (MW = 284.48 and main MS nger prints: 74, 143, 255, 298m/z); 18:1 (MW = 282.52 and main MS
nger prints: 55, 74, 97, 264m/z); and 18:2 (MW = 278.43 and
main MS nger prints: 55, 81, 109, 294m/z), as shown in MS boxes
in Fig. 2. Relative composition of the main fatty acids in wastewater sludge were: myristic (4.6%), palmitic (37.6%), stearic (17.6%),
palmitoleic (4.8%), oleic (19.5%), and linoleic (6.3%). This distribution constitutes the main FAME components dominating common
biodiesel feedstocks (Schober et al., 2006). Characterization of municipal wastewater sludge fatty acid composition by GCMS adds
to the reliability of previous qualitative GC analysis conducted on
these bio-wastes as suitable lipid-feedstocks for biodiesel
production.
Gas chromatograph fatty acid proles of individual waste
streams are presented in Fig. 3. Fatty acid compositions of olive
mill pomace and wastewater are illustrated in Fig. 3a, and of primary, secondary, and stabilized sludge in Fig. 3b. The source of
the two oils differs signicantly: olive mill waste oil originates
from pure vegetable oil (Kiritsakis, 1998); while municipal wastewater sludge oil originates from a mixture comprised of microorganisms among other organic lipids (Fytili and Zabaniotou,
2008). Both waste oils have in common the main fatty acid groups
of C16 and C18, as can be observed in Fig. 3a and b. OMP and OMW
fatty acid proles presented in Fig. 3a exhibit similar fatty acid
compositions, dominated by the oleic fatty acid (>60%). The great
similarity between the two olive mill waste oils enabled TD-NMR
analysis of the wastes using one representative calibration
(Fig. 1a). The main fatty acids found in olive mill waste oils were:
stearic (11.0%), palmitoleic (12.7%), oleic (61.0%), and linoleic
(10.8%). Fatty acid composition of olive mill pomace and wastewa-

Fig. 2. GCMS fatty acid prole of municipal wastewater sludge (primary, secondary, and stabilized sludge) integrated with mass spectrometer identication ngerprints of
main fatty acids present.

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R.M. Willson et al. / Waste Management 30 (2010) 18811888

Fig. 3. (a) GC fatty acid prole comparison of olive mill pomace and wastewater. (b) GC fatty acid prole comparison of primary, secondary, and stabilized municipal
wastewater sludge.

ter show strong similarity to virgin olive oil, indicating high quality
and stability of waste oils (Kiritsakis, 1998).
Fatty acid proles of primary, secondary, and stabilized sludge,
presented in Fig. 3b, enabled comparison between the fatty acid
compositions of the various sludge types produced during activated-sludge wastewater treatment. This comparison constitutes
an innovation in the characterization of municipal wastewater
sludge lipid fraction. Prole comparison between the three sludge
types revealed strong similarity in their fatty acid compositions. In
the context of TD-NMR analysis, the similarity in fatty acid composition of the three sludge types enabled analysis of wastewater
sludge samples by TD-NMR using one representative calibration
(Fig. 1a). Primary, secondary, and stabilized sludge all have a large
palmitic fatty acid content, ranging from 35% to 40%, followed by
stearic and oleic fatty acids ranging from 15% to 25% (Fig. 3b). Large
content of these fatty acids, which are abundant in typical vegetable oil biodiesel feedstocks, shows the waste oils potential suitability for biodiesel production (Schober et al., 2006). Most
previous research on this subject has focused on one sludge type
due to the complex lipid fraction present in sewage sludge. Recently Mondala et al. (2009) conducted a prole comparison between primary and secondary sludge. Their ndings show similar
FAME compositions for primary and secondary sludge, dominated
by palmitic, oleic, and stearic fatty acids.
Typical fatty acid composition of olive mill waste and municipal
wastewater sludge were compared to common biodiesel feedstocks on the basis of fatty acid saturation level, as shown in
Fig. 4. Olive mill waste fatty acid composition shows strong similarity to rapeseed oil due to a high content of mono-unsaturated
fatty acids in both vegetable oils. Both olive mill waste and rape-

seed oil contain over 60% oleic acid (C18:1), which has been proven
to be a benecial feedstock for biodiesel production since it exhibits a combination of improved fuel properties (Knothe, 2005).
Moreover, total unsaturation levels in olive mill waste oil are very
similar to those of soy and rapeseed oil (84.5% versus 91.5% in soy
and 83.8% in rapeseed oil; Chapagain et al., 2009). This similarity
also implies a potential compatibility of olive mill waste oils with
international biodiesel standards. Moreover, recent research by
Dorado et al. (2003) found the fuel properties of cooking-waste

Fig. 4. Fatty acid prole comparison of olive mill waste and municipal wastewater
sludge to soy and rapeseed standard biodiesel oil feedstocks based on saturation
level. Fatty acid prole of all oils was carried out in the Phyto-Lipid Biotechnology
Laboratory, at BGU.

R.M. Willson et al. / Waste Management 30 (2010) 18811888

olive oil methyl esters, with properties similar to those of olive mill
waste oils, compatible with European FAME standards.
The fatty acid composition of municipal wastewater sludge presented in Fig. 4 represents an average of primary, secondary, and
stabilized sludge. This typical sewage sludge fatty acid composition is slightly different from the three vegetable-based oils due
to higher levels of saturation (Fig. 4). Generally, it is expected that
high levels of saturated fatty acids may yield a relatively high viscosity in the obtained oil and later in the produced biodiesel (Doshi
et al., 2005). However, a study by Knothe (2005) has shown that a
decrease in the chain length, as shown in the case of municipal
wastewater sludge oil (Figs. 2 and 3b), dominated by palmitic acid
(16:0, >35%), leads to decreased viscosity, melting point, cetane
number, and heat of combustion.
Fatty acid compositions of both wastes indicate that the waste
oils are of good quality and have an intact FAME composition.
Greater levels of saturation reduce susceptibility to oxidation,
increasing stability of the oils and therefore the derived biodiesel
(Chapagain and Wiesman, 2009). Since the present and near future
international regulations allow only blended biodiesel such as B5,
B10, or B20 (US standard: ASTM, 2007; EU regulations: EN
14214, 2003), there is a strong chance that these potential biowaste oil feedstock resources may meet the required international
standards. Biodiesel production limitations of current vegetable oil
feedstocks have resulted in multi-feedstock processing of almost
all types of fats and oils into biodiesel, which has become possible
due to recent development of new production technologies (Schober et al., 2006). As a result, current strict biodiesel standards will
become more exible to the qualitative perspectives of alternative
oil feedstocks as recently proposed by Schober et al. (2006).
3.4. Biodiesel potential of olive mill waste and sewage sludge in Israel
Statistics on current waste generation in Israel along with their
oil and biodiesel potential are presented in Table 2. The Israeli olive
mill industry currently generates 120,000 tons of waste per annum. Based on oil and biodiesel yields obtained in this research,
an estimated 2800 tons of oil, which may produce 2500 tons of biodiesel per annum in Israel, is currently wasted. Based on current
wastewater generation in Israel, a total of 18 Mm3/year of primary
and secondary sludge may be potentially generated (www.sviva.com/olive/mill wastewater). Primary and secondary sludge are

Table 2
Estimation of annual oil and biodiesel potential of olive mill wastes and municipal
wastewater sludge generated in Israel.
Waste
generated in
Israel

Waste generation
(1000 tons/year)
Total
weight

(a) Olive mill waste

Pomace
30
Wastewater
90

Dry
material
15.3
6.7

(b) Municipal wastewater sludge

Primary sludge 9000a
400
Secondary
9000a
100
sludge
500
100
Stabilized
sludgeb
Total

18,120

Oil potential
(1000 tons/year)

522

Biodiesel
potential
(1000 tons/
year)

(%)

1.7
1.1

1.5
1

2.4
1.6

92
18

46
9

71
14

14

11

112.8

57.5c

100

1887

further processed to produce an estimated 500,000 m3/year of

semi-solid stabilized sludge. Based on the present research, an estimated 92,000 and 18,000 tons of oil may be derived from primary
and secondary sludge, respectively, or 14,000 tons of oil from
anaerobically stabilized sludge. The annual biodiesel potential of
Israel municipal wastewater sludge was estimated at 46,000 and
9000 tons derived from primary and secondary sludge, or an estimated 7000 tons of biodiesel from anaerobically stabilized sludge.
These gures reveal a large potential feedstock for future biodiesel
production in Israel (Table 2). Evaluation of olive mill waste and
municipal wastewater sludge biodiesel potential may be conducted on these wastes and others generated in signicantly larger
quantities across the world. Large quantities and high oil content
show the potential of bio-wastes as favorable feedstock for future
expansion of the biodiesel industry in answer to the growing demand for its production.
A preliminary economic evaluation of the nancial biodiesel potential from olive mill waste and municipal wastewater sludge in
Israel was conducted. Currently a well-developed biodiesel industry does not exist in Israel due to lack of viable feedstocks for biodiesel production (www.sviva.com/energy). Therefore, the
potential biodiesel market of the wastes was based on current
international pricing of soy oil-based biodiesel. A potential market
value of $6977 million US was predicted for reutilization of olive
mill waste and municipal wastewater sludge in Israel for biodiesel
production. These predictions should increase over time as the
supply of diesel from conventional sources is depleted. Moreover,
the potential market value has not taken into account nancial
and environmental benets of reutilizing wastes as a source for
alternative clean energy. The most benecial waste source for
biodiesel production was primary sludge consisting over 70% of
the total biodiesel market value among the ve Israeli waste
sources evaluated (Table 2). Biodiesel production potential from
the two agro-industrial wastes tested may replace 3% of current
diesel consumption in Israel, further reinforced by the many benets of blending biodiesel with conventional diesel at up to 5%
which results in improved fuel properties (Due et al., 2008).
4. Conclusions
Olive mill waste and municipal wastewater sludge contain substantial amounts of oil, based on conventional analyses complimented by TD-NMR analysis. TD-NMR has been demonstrated as
a unique tool that can be used accurately, cost effectively, and
quickly to identify various waste-oil sources and support new biodiesel industries. Specic fatty acid analyses of the oils demonstrate their suitability for potential biodiesel production. It was
clearly shown that the two bio-waste sources tested can become
valuable oil sources for biodiesel industry expansion to new regions such as Israel. Biomass waste based alternative energy will
also reduce solid waste accumulation, addressing challenging environmental waste treatment and disposal problems. However, still
further work of industrial scaling up is needed.
Acknowledgments
The authors wish to thank Dr. Bishnu Chapagain, Mr. Shahar
Nizri, and Ms. Shirley Berman for their efcient and kind technical
support. This study was nancially supported by a grant from Chief
Scientist of the Israeli Ministry of Infrastructure.

Based on sludge treatment of all wastewaters produced in Israel

(www.sviva.com).
b
Produced by further processing of primary and secondary sludge, which are
more benecial feedstock for biodiesel production and therefore deducted from
total calculations.
c
Assuming oil to biodiesel conversion efciency of about 50%.

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