Energy Fuels 2010, 24, 1037–1042 : DOI:10.

1021/ef9011388
Published on Web 01/14/2010

Molecular and Isotopic Analysis of Motor Oil from a Biodiesel-Driven Vehicle

Emily E. Peacock,*,† J. Samuel Arey,‡ Jared A. DeMello,† Ann P. McNichol,§ Robert K. Nelson,† and
Christopher M. Reddy†
†
Department of Marine Chemistry and Geochemistry, Woods Hole Oceanographic Institution, Woods Hole, Massachusetts 02543,
‡
Environmental Chemistry Modeling Laboratory, Swiss Federal Institute of Technology at Lausanne, 1015 Lausanne, Switzerland,
and §National Ocean Sciences Accelerator Mass Spectrometry Facility, Department of Geology and Geophysics, Woods Hole
Oceanographic Institution, Woods Hole, Massachusetts 02543

Received October 7, 2009. Revised Manuscript Received December 20, 2009

Biodiesel, a mixture of fatty acid methyl esters (FAMEs), is increasingly recognized as a renewable fuel.
While some environmental impacts of biodiesel usage have been investigated, accumulation of organic
compounds in motor oil, which can subsequently leak onto roads, has not been studied. Because studies
have shown that toxic polycyclic aromatic hydrocarbons (PAHs) accumulate in the motor oil of engines
fueled with fossil diesel or gasoline, the objective of this study was to determine if this also occurs for
engines fueled with biodiesel. Here, we sampled and analyzed motor oil of a biodiesel-powered 2005
Volkswagen Passat Wagon over 3240 km of personal-use driving. Using gas chromatography with flame
ionization detection (GC-FID), we found a total of 0.5% FAMEs in the motor oil after 3240 km. We also
used gas chromatography-mass spectrometry and comprehensive two-dimensional gas chromatography
and did not detect PAHs or other organic compounds not present in the initial motor oil. Using natural
radiocarbon analysis, a powerful technique capable of detecting biodiesel-derived carbon that would be
otherwise undetectable by gas chromatography, we found a total of 0.68% biodiesel-derived carbon after
3240 km. This is similar to the amount of FAMEs found in these samples with GC-FID, indicating that
the primary source of biodiesel-derived carbon in the motor oil is FAMEs (and not PAHs or other
carbonaceous species). This result suggests that used motor oil of biodiesel vehicles can be less toxic based
on PAH content than vehicles fueled with fossil diesel or gasoline.

Introduction use associated with producing biodiesel,3,9-12 but efforts are

Biodiesel is a mixture of fatty acid methyl esters (FAMEs)
derived from animal fats or vegetable oils. The environmental and
health impacts of the biodiesel lifecycle have received considerable
attention over the last 2 decades.1-3 Large-scale implementation
of biodiesel continues to be of great interest because of its potential
to be a “low carbon fuel” that is nearly interchangeable with fossil
diesel,2 usually with a minimal decrease in engine power and
increase in fuel consumption.1,4-6 It also presents an opportunity
to lessen the dependence upon foreign oil imports for many
countries.7,8 Some concerns exist about excess water and land
*To whom correspondence should be addresssed. Telephone:
(508)-289-3568. Fax: (508) 457-2134. E-mail: epeacock@whoi.edu.
(1) Basha, S. A.; Gopal, K. R.; Jebaraj, S. Renewable Sustainable
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Third Liquid Fuel Conference, Liquid Fuels and Industrial Products from
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Press: Urbana, IL, 2005; pp 17-25.
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Environ. Sci. Technol. 2009, 43, 3005–3010.
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underway to develop alternative sources and production techni-
ques.13-16 Hence, understanding the environmental impacts of
biodiesel usage continues to warrant study.
To evaluate the environmental impacts of this alternative
fuel in the context of factors commonly associated with the use
of traditional fossil diesel, it is prudent to at least consider
terrestrial and marine spills, biodegradability, exhaust emis-
sions, and accumulation of toxic compounds in motor oil.
Biodiesel biodegrades rapidly in seawater and rainwater rela-
tive to fossil diesel.17,18 Numerous studies have focused on the
regulated exhaust emissions associated with biodiesel combus-
tion, with the general result of a reduction of most emissions
but an increase in NOx gases.7,19,20 In one study of biodiesel
use under low-speed, urban conditions and in engines that are
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Elobeid, A.; Fabiosa, J.; Tokgoz, S.; Hayes, D.; Yu, T.-H. Science
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r 2010 American Chemical Society 1037 pubs.acs.org/EF

Energy Fuels 2010, 24, 1037–1042 : DOI:10.1021/ef9011388
optimized for petroleum diesel, it was found that regulated
exhaust emissions increased relative to emissions observed with
petroleum diesel use.21 However, the combined results of recent
studies affirm that more work is needed regarding emission
levels, especially with respect to toxicity.22,23 For example,
particle extracts of exhaust emissions from biodiesel usage
exhibit increased cytotoxicity or mutagenicity compared to
those from fossil diesel exhaust.24,25 Finally, the analysis of
motor oil for toxic organic compounds has yet to be studied for
biodiesel-operated vehicles.
For traditional diesel and gasoline engines, numerous
studies have shown that used motor oil contains toxic and
mutagenic polycyclic aromatic hydrocarbons (PAHs) that
accumulate steadily with mileage. This is due to direct leakage
of fuel into the motor oil as well as the accumulation of
incomplete combustion products.26,27 For gasoline-driven
vehicles, levels as high as 10 000 μg g-1 or about 1% of fuel-
derived PAHs have been measured in used motor oil.26-31
Diesel motor oils also accumulate PAHs, although typically at
lower amounts than gasoline-driven vehicles.26,32 Used motor
oil is a main contributor of PAHs to urban runoff and,
consequently, to the aquatic environment.33-39 Thus, as a
new fuel enters the market, it is important to determine what
organic compounds, including PAHs, will accumulate in
motor oil from biodiesel-operated vehicles.
Research on used motor oil from engines operating with
biodiesel has focused primarily on the accumulation of metals
as an indication of engine wear. Replacing fossil diesel with
biodiesel reduces engine wear as well as the accumulation of
aluminum, iron, chromium, and lead in the motor oil.5,40-43 In
addition, a number of studies from the early 1980s examined
(21) Fonteras, G.; Karavalakis, G.; Kousoulidou, M.; Tzamkiozis,
T.; Ntziachristos, L.; Bakeas, E.; Stournas, S.; Samaras, Z. Fuel 2009, 88,
1608–1617.
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Perspect. 2007, 115, 496–499.
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Res. 2009, 9, 18–31.
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M. C.; Ghio, A. J. Open Toxicol. J. 2009, 3, 8–15.
(25) Krahl, J.; Knothe, G.; Munack, A.; Ruschel, Y.; Schr€ oder, O.;
Hallier, E.; Westphal, G.; B€ unger, J. Fuel 2009, 88, 1064–1069.
(26) Grimmer, G.; Jacob, J.; Naujack, K. W.; Dettbarn, G. Fresenius’
J. Anal. Chem. 1981, 309, 13–19.
(27) Pruell, R. J.; Quinn, J. G. Environ. Pollut. 1988, 49, 89–97.
(28) Payne, J. F.; Martins, I. Science 1978, 200, 329–330.
(29) Clonferno, E.; Nardini, B.; Marchioro, M.; Bordin, A.; Gabbani,
G. Mutat. Res. 1996, 368, 283–291.
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(31) Lu, S. T.; Kaplan, I. R. Environ. Forensics 2008, 9, 295–309.
(32) Carmichael, P. L.; Jacob, J.; Grimmer, G.; Phillips, D. H.
Carcinogenesis 1990, 11, 2025–2032.
(33) Hunter, J. V.; Sabatino, T.; Gomperts, R.; MacKenzie, M. J.
J. Water Pollut. Control Fed. 1979, 51, 2129–2173.
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Technol. 1981, 15, 315–326.
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Quinn, J. G. Water, Air, Soil Pollut. 1990, 52, 1–21.
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310–315.
(38) National Research Council. Oil in the Sea III: Inputs, Fates, and
Effects; The National Academies Press: Washington, D.C., 2003.
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Sci. Technol. 1984, 18, 580–587.
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changes in motor oil viscosity using experimental bench en-
gines.44-47 In a 1994 study comparing fossil diesel and biodiesel
(B100) in bus engine oil, it was found that biodiesel accumu-
lated in bus motor oil to a much higher level (9.5%) than fossil
diesel (2.5%) after about 10 000 km of driving.48 More recently,
a bench engine experiment using B100 exhibited only a fuel
accumulation of 3.5% in the used motor oil after 150 h of
operation.49
Here, we investigated whether organic compounds accumu-
late in the motor oil of a modern, biodiesel-fueled car. Our
motivation was to determine whether motor oils from biodiesel-
driven cars could become a major PAH (as well as other
organic compounds) input term to the aquatic environment,
presuming that the usage of this fuel continues to increase. In
addition, our hope was that this study may shed light on the
performance of modern engines fueled by biodiesel.
Experimental Section
We employed an approach similar to that of Pruell and
Quinn,27 who sampled the oil of a 2 1/2-year-old Mazda GLC
gasoline-fueled automobile and found that PAHs increased with
mileage following an oil change. Here, we analyzed the motor oil
of a biodiesel-fueled car for FAMEs and PAHs via gas chromato-
graphy with flame ionization detection (GC-FID) and mass
spectrometry (GC-MS), respectively. We also analyzed the
initial and final samples on comprehensive two-dimensional gas
chromatographs with either a flame ionization detector (GC

GC-FID) or time-of-flight mass spectrometry (GC
GC-MS).
Finally, building on previous work, we analyzed select samples
for natural radiocarbon (14C; half-life of 5730 years)50-52 to
quantify the total biodiesel-derived carbon in the motor oil,
including components that may be undetected via gas chroma-
tography. This approach relies on differences in the 14C abun-
dance in motor oil and biodiesel.52 Motor oil, derived from fossil
carbon, contains no detectable 14C. Biodiesel has contemporary
or modern levels of 14C.52 Hence, 14C is an ideal tracer for
biodiesel in motor oils because it is effectively prelabeled with 14C.
We began this study with an oil change of a 2005 Volkswagen
Passat Wagon (2.0 L engine; 18 805 km driven) using an Elf
5W-40 synthetic motor oil, the only product used in this vehicle.
We then collected oil samples during a total of 3240 km of routine
driving in the Los Angeles region of California. The current
owner purchased the car when it had 3494 km and since then had
used only B99.9. During the course of the study, the owner used
only B99.9 from one fueling station, and fuel was collected for
analysis at three time points. Because B99.9 gels at ∼3 °C,53 we
chose to perform this experiment in a warm climate, where average
temperatures for the study time period are between 18 °C (low) and
(44) Fort, E. F.; Blumberg, P. N.; Staph, H. E.; Staudt, J. J. SAE Tech.
Pap. 820317, 1982.
(45) Pischinger, G. H.; Seikman, R. W.; Falcon, A. M.; Fernandez,
F. R. Proceedings of the 5th International Symposium on Alcohol Fuels
and Technology, Auckland, New Zealand, 1982.
(46) Siekmann, R. W.; Pischinger, G. H.; Blackman, D.; Carvalho,
L. D. Proceedings of the International Conference on Plant and
Vegetable Oils as Fuels, American Society of Agricultural Engineers
(ASAE), Fargo, ND, 1982.
(47) Blackburn, J. H.; Pinchin, R.; Nobre, J. I. T.; Crichton, B. A. L.;
Cruse, H. W. SAE Tech. Pap. 831355, 1983.
(48) Sadeghi-Jorabchi, H.; Wood, V. M. E.; Jeffery, F.; Bruster-Davies,
A.; Loh, N.; Coombs, D. Spectrosc. Eur. 1994, 6, 16–20.
(49) Y€ €
uksek, L.; Kaleli, H.; Ozener, € guz, B. FME Trans. 2009,
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37, 91–97.
(50) Bennett, M.; Volckens, J.; Stanglmaier, R.; McNichol, A. P.;
Ellenson, W. D.; Lewis, C. W. Aerosol Sci. 2008, 39, 667–678.
(51) Cheng, A. S.; Buchholz, B. A.; Dibble, R. W. SAE Tech. Pap.
2003-01-2283, 2003.
(52) Reddy, C. M.; DeMello, J. A.; Carmichael, C. A.; Peacock, E. E.;
Xu, L.; Arey, J. S. Environ. Sci. Technol. 2008, 42, 2476–2482.
(53) Joshi, M. R.; Pegg, M. J. Fuel 2007, 86, 143–151.
Energy Fuels 2010, 24, 1037–1042 : DOI:10.1021/ef9011388 Peacock et al.

Table 1. Isotopic Composition of Motor Oil and Select Samples of Fuel Used in the Study
distance after NOSAMSa
sample type date oil change (km) δ13C (%) Δ14C (%) accession number
motor oil sampled motor oil 7/4/2006 0 -29.0 -996 OS-66205
from a 2005 motor oil 7/7/2006 188 nab na na
Volkswagen Passat Wagon motor oil 7/8/2006 591 -29.0 -994 OS-67370
motor oil 7/10/2006 787 na na na
motor oil 7/26/2006 1556 -29.0 -993 OS-67371
motor oil 8/3/2006 1868 na na na
motor oil 8/12/2006 2210 -29.0 -992 OS-67369
motor oil 8/14/2006 2634 na na na
motor oil 8/23/2006 2989 na na na
motor oil 9/4/2006 3240 -29.0 -989 OS-66206

fuels used during experiment B99.9 fuel 7/4/2006 0 -28.9 7.7 OS-56773
B99.9 fuel 7/8/2006 591 -28.6 9.7 OS-56774
B99.9 fuel 8/16/2006 2797 -27.8 11 OS-56775
a
National Ocean Sciences Accelerator Mass Spectrometry Facility located at the Woods Hole Oceanographic Institution, Woods Hole, MA. b Not
analyzed.

29 °C (high). The motor oil was sampled a total of 10 times,
starting immediately after the oil change during July, August,
and September of 2006 (Table 1). Each sample was collected
when the engine was cold using the dipstick of the engine to
transfer the oil directly to precombusted 4 mL amber glass vials
with solvent-cleaned Teflon-lined caps. All samples were ship-
ped to our laboratory in Woods Hole, MA, and stored at 1 °C
until analysis. Samples were prepared for gas chromatographic
analysis by dissolving 25 mg of sample into 5 mL of methyl
tert-butyl ether (MTBE). In addition, both the original and final
(after 3240 km of driving) motor oil samples were esterified in
methanolic HCl to check for any changes in the amount of free
fatty acids.
We quantified FAMEs using a Hewlett-Packard 5890 Series II
GC-FID. A 0.5 μL sample was injected cool-on-column. Com-
pounds were separated on a glass capillary column (J&W DB-1MS,
30 m, 0.25 mm inner diameter, 0.25 μm film) with H2 as the
carrier gas at a constant flow of 5 mL min-1. The GC oven
temperature was programmed from 45 °C (5 min hold) to 315 °C
at 7 °C min-1 to 320 °C at 20° min-1 (10 min hold). Samples were
spiked with an internal standard hexadecane-d34, and FAMEs
were quantified using previously published methods.17
We checked the identity of the individual FAMEs with two
standards purchased from Supelco, FAME Mix Rapeseed Oil
Lot Number LB-34981 and FAME Mix RM-6 Lot Number
LB-37097. The estimated method detection limit for individual
FAMEs was 0.05 μg g-1 oil. Solvent blanks were free of FAMEs.
Precision of FAMEs content based on duplicate analysis was
about 10%. To test the performance of the method, FAMEs were
spiked into synthetic motor oil. The recoveries of the FAMEs
were greater than 95% (we employed a similar approach to
evaluate PAH analysis).
PAHs were measured with an Agilent 6890 series gas chromato-
graph with an Agilent 5973 mass selective detector (GC-MS).
The instrument was operated in full-scan mode ranging from
50 to 800 amu. Compounds were separated on a J&W DB-XLB
capillary column (60 m, 0.25 mm inner diameter, 0.25 μm film)
with He as the carrier gas at a constant flow of 1.5 mL min-1. The
GC oven had an initial temperature of 50 °C (1 min hold) and
was ramped at 20 °C min-1 to 115 °C (10 min hold) and then at
5 °C min-1 to 320 °C (10 min hold). Samples were spiked with the
following standards: naphthalene-d8, fluorene-d10, acenaphthene-
d10, phenanthrene-d10, dibenzothiophene-d8, fluoranthene-d10,
chrysene-d10, and perylene-d12. We then analyzed for the following
parent PAHs and their alkylated homologues: naphthalene, phe-
nanthrene/anthracene, fluorene, dibenzothiophene, fluoranthene,
pyrene, chrysene, benz[a]anthacene, benzofluoranthenes, benzo-
[e]pyrene, benzo[a]pyrene, and perylene. Method detection limits,
estimated from replicate analysis of PAH-containing bunker fuel
using the approach of Glaser et al.,54 ranged from 0.004 to 0.05 μg
1039
of PAH g-1 of oil. Solvent blanks were free of PAHs and other
petroleum hydrocarbons. On the basis of replicate analysis, preci-
sion of PAH content in oil was 10% or lower. Recoveries of PAHs
spiked into the synthetic motor oil were greater than 90%.
We analyzed the initial and final motor oil samples on a Leco
GC
GC-FID system that employed a dual-stage cryogenic
modulator installed in an Agilent 7890A gas chromatograph
configured with a 7683 series split/splitless autoinjector, two
capillary gas chromatography columns, and a flame ionization
detector. The extract was injected in splitless mode, and the purge
vent was opened at 1.0 min. The inlet temperature was 300 °C.
The first-dimension column and the dual-stage cryogenic modu-
lator reside in the main oven of the Agilent 7890A gas chromato-
graph. The second-dimension column is housed in a separate
oven installed within the main GC oven. The first-dimension
column was a nonpolar Restek Rtx-1MS (19 m, 0.2 mm inner dia-
meter, 0.2 μm film), programmed to remain isothermal at 110 °C
(10 min hold) and then ramped from 110 to 330 °C at 1.5 °C min-1.
Compounds eluting from the first-dimension column were cryo-
genically trapped and re-injected (modulated) onto the second-
dimension column at a period of 5 s. The modulator cold jet gas
was dry N2, chilled with liquid N2. The thermal modulator hot jet
air was heated to 35 °C above the temperature of the main GC
oven (thermal modulator temperature offset = 35 °C). The hot
jet was pulsed for 0.33 s every 5 s, with a 2.17 s cooling period
between stages. Second-dimension separations were performed
on a 50% phenyl polysilphenylene-siloxane column (SGE
BPX50, 1 m, 0.10 mm inner diameter, 0.1 μm film) that was
programmed to remain isothermal at 125 °C (10 min hold) and
then ramped from 125 to 345 °C at 1.5 °C min-1. The carrier gas
was H2 at a constant flow rate of 0.95 mL min-1. The FID
detector signal was sampled at 100 Hz.
We also analyzed the initial and final motor oil samples on a Leco
GC
GC time-of-flight-mass spectrometer. GC
GC-MS oven
temperature conditions were identical to the GC
GC-FID, with
the following exceptions: the main GC oven on the GC
GC-MS
system is a 6890N; the carrier gas was He; and the first-dimension
column was a Restek Rtx-1MS (25 m, 0.2 mm inner diameter,
0.2 μm film). The MS detector signal was sampled at a data rate of
50 Hz. The transfer line from the second oven to the MS was
deactivated fused silica (0.5 m, 0.18 mm inner diameter) that was
held at a constant temperature of 275 °C. The MS source tempera-
ture was 225 °C; the detector voltage was 1575 V; the mass scan
range was 50 to 650 amu; and the MS mass defect was manually set
at 98.6 amu 100 units-1. The mass spectrometer was set at 70 eV
electron ionization and with a push-pulse rate of 5 kHz.
(54) Glaser, J. A.; Foerst, D. L.; McKee, G. D.; Quave, S. A.; Budde,
W. L. Environ. Sci. Technol. 1981, 15, 1426–1435.
Energy Fuels 2010, 24, 1037–1042 : DOI:10.1021/ef9011388
Figure 1. Time series of GC-FID chromatograms of the five engine oil samples that were also analyzed for 14C: (a) 0 km, (b) 591 km, (c) 1556 km,
(d) 2210 km, and (e) 3240 km. The peak marked with an asterisk is the internal standard. The boxed area in each panel is an expanded view of the
FAMEs that are labeled in e.
Finally, we analyzed the fuel samples and select motor oil
samples for stable carbon and radiocarbon content using methods
previously described in detail.52,55 Briefly, each sample was quanti-
tatively converted from organic carbon into CO2 and then into
graphite for isotopic analysis. On a small fraction of the CO2, the
stable carbon isotope ratios (δ13C) were determined via isotope
ratio mass spectrometry. The 14C content of the graphite was
measured by accelerator mass spectrometry (AMS) at the National
Ocean Sciences Accelerator Mass Spectrometry (NOSAMS) faci-
lity in Woods Hole, MA.55 In this study, all 14C measurements are
expressed with the Δ14C nomenclature, as defined in Stuiver and
Polach;56 Δ14C expresses the deviation of the radiocarbon content
of a sample, normalized to a constant δ13C (-25%) and time
(1950), from “modern” as defined by the international radiocarbon
community. With this nomenclature, fossil carbon and modern
carbon have Δ14C values of -1000 and >0%, respectively. The
precision and accuracy of these measurements are discussed in
detail elsewhere.52
(55) McNichol, A. P.; Osbourne, E. A.; Gagnon, A. R.; Fry, B.; Jones,
G. A. Nucl. Instrum. Methods Phys. Res., Sect. B 1994, 92, 162–165.
(56) Stuiver, M.; Polach, H. A. Radiocarbon 1977, 19, 355–363.
1040
Peacock et al.
Results and Discussion
In this study, we investigated the accumulation of organic
compounds in motor oil from a biodiesel-driven car starting
at the time of an oil change and sampling periodically over
3240 km (Table 1).
In the GC-FID chromatograms, the biodiesel FAMEs
elute between the n-C19 and n-C22 alkane elution window and
prior to three humps (panels a-e of Figure 1). These humps or
unresolved complex mixtures residing within the elution
window of n-C23 and n-C43 alkanes are consistent with
published chromatograms of Elf 5W-40 synthetic oil;57 their
relative abundance and distribution did not change through-
out the study. However, the amount of FAMEs in the motor
oil increased steadily with mileage from 0.1 to 0.5% during the
course of the study (Figures 1 and 2) [in the initial sample, we
assume that the FAMEs content (0.1%) was a carryover of
the residual oil from the previous change]. The final amount of
(57) Mao, D.; Lookman, R.; Vanermen, G.; De Brucker, N.; Diels, L.
Fuel 2009, 88, 312–318.
Energy Fuels 2010, 24, 1037–1042 : DOI:10.1021/ef9011388
Figure 2. Plots of changes in chemical composition versus the
distance traveled. (a) FAMEs measured by GC-FID (all samples)
and (b) 14C content (select samples). Note that the 14C approach
does not have an estimate for the beginning of the study because we
used the Δ14C value of the initial motor oil as an end member.
FAMEs is much lower than levels published in previous
efforts. For example, in one study using a bench engine
running at full power burning 100% fatty acid ethyl esters
(FAEEs), the motor oil was over 20% fuel at the end of the
experiment.47 A more recent study employing a bench engine
combusting biodiesel found a 3.5% accumulation of FAMEs
in the motor oil after 150 h of engine operation.49 It is difficult
explain these differences compared to our results; presumably,
numerous factors may be responsible, including the study
conditions or engine performance.
Further analysis of GC-FID data provided additional
information about the FAMEs in the motor oil. The FAMEs,
identified in Figure 1e, include methyl hexadecanoate
(C16 FAME), methyl octadecanoate (C18 FAME), and C18
FAME isomers with one, two, or three double bonds refer-
red to as C18:1, C18:2, or C18:3 FAMEs, respectively. The rela-
tive distribution of the FAMEs was nearly constant over
3240 km but differed from the three fuel samples, which had
similar distributions, taken during the sampling period
(results not shown). Comparing the fuel to the motor oil, we
observed a relative increase in the proportion of C16:0 and
C18:0 FAMEs and a decrease in C18:2 FAME, indicating the
preferential loss of unsaturated FAMEs, likely because of
oxidation. Because we did not sample B99.9 at every fill up, we
are hesitant to make further inferences based only on
GC-FID data. To test for any free fatty acids buildup with
usage, we esterified the initial and final oil sample. No increase
in the overall FAME content of the motor oils was observed.
1041
Peacock et al.
Hence, there was no detectable generation or accumulation of
free fatty acids.
GC-MS analysis of the motor oil samples revealed that
there were no PAHs present at levels above our method
detection limits. This indicates that replacing fossil diesel with
biodiesel can lead to less input of toxic compounds to the
environment from leaking oil as well as improper disposal.
The presence of FAMEs is viewed without any major con-
cerns because they are not harmful and have been shown
to biodegrade readily in laboratory studies.17,18,58 Taken
together, the presence of FAMEs and absence of PAHs
suggests that the motor oil composition changed because of
the direct dilution by fuel during operation. This is consistent
with previous work on gasoline-fueled vehicles. Grimmer
et al.26 found that the major portion of the PAH accumulation
in the motor oil of a gasoline-powered car originated from the
fuel and not incomplete combustion that could produce
PAHs. To evaluate our PAH data, we considered the follow-
ing calculations. At the end of the study, the used motor oil
had 5 mg g-1 of FAMEs (total biodiesel) in the motor oil.
Because the fuel did contain 0.1% fossil diesel, which un-
doubtedly also leaked into the motor oil, then it is reasonable
that PAHs from this leakage also occurred. Generally, total
PAHs are about 5% for a fossil diesel.59 Then, the resulting
PAH content in the motor oil would be 5% of 5 μg g-1 or
250 ng g-1. However, because that amount of total PAHs is
split among many individual compounds (∼25),59 we calcu-
late that each compound would be ∼0.01 μg g-1 in the used
motor oil. Such levels are at or below our calculated method
detection limits. This would imply that, when used with
biodiesel, motor oil contains PAH levels that are 2-3 orders
of magnitude lower than reported values for PAH levels in
used diesel motor oils26,32 and 4-5 orders of magnitude
smaller than PAH levels in used gasoline motor oil, which
were 100-1000 μg g-1.26,27
GC
GC further confirmed results found using GC-FID
and GC-MS. The initial and final samples were analyzed by
GC
GC-FID and GC
GC-MS (Figure 3; GC

GC-FID images only shown). In comparison to traditional
gas chromatography, this technology separates and resolves
at least an order of magnitude more compounds, has a much
larger signal-to-noise ratio, and sorts compounds based on
their chemical class; hence, it provides highly refined inven-
tories of organic compounds in complex mixtures.60 GC

GC chromatograms confirmed the accumulation of FAMEs
and absence of PAHs (and other organic compounds) in the
used motor oil.
To determine whether any breakdown products of com-
busting biodiesel, not detected by GC, were present in the
motor oil samples, we measured Δ14C values for five select
samples (Table 1). The 14C content in the motor oil samples
increased with time and mileage, exhibiting Δ14C values of
-996.0 ( 0.4% at the time of the oil change and -989.2 (
0.4% after 3240 km of driving. These values indicate that
most of the carbon in the sample has a fossil origin, which
(58) Owsianiak, M.; Chrzanowski, Le.; Szulc, A.; Staniewski, J.;
Olszanowski, A.; Olenjnik-Schmidt, A. K.; Heipieper, H. J. Bioresour.
Technol. 2009, 100, 1497–1500.
(59) Stout, S. A.; Uhler, A. D.; McCarthy, K. J.; Emsbo-Mattingly, S.
Chemical fingerprinting of hydrocarbons. In Introduction to Environ-
mental Forensics; Murphy, B. L., Morrison, R. D., Eds; Academic Press: San
Diego, CA, 2002; pp 137-260.
(60) Reddy, C. M.; Eglinton, T. I.; Hounshell, A.; White, H. K.; Xu,
L.; Gaines, R. B.; Frysinger, G. S. Environ. Sci. Technol. 2002, 36, 4754–
4760.
Energy Fuels 2010, 24, 1037–1042 : DOI:10.1021/ef9011388
Figure 3. GC
GC-FID images of the (A) initial and (B) final
engine oil samples. A volatility-based separation with a nonpolar
column occurs along the x axis. A polarity-based separation with a
semi-polar column yields chemical class-type separation along the
y axis. The FAMEs, hopanes, which are chemical fossils found in
synthetic motor oil, and bulk motor oil peaks are annotated. No
other peaks appear in the final sample relative to the initial sample
besides the FAMEs. These chromatograms also are consistent with
previous studies on Elf motor oil.57
would again be -1000%, reflecting that the sample is predomi-
nantly composed of synthetic motor oil. By comparison, the
B99.9 fuels used in the car and collected from fueling stations 3
times during the experiment had values Δ14C ranging from 7.7 to
11% (Table 1). The δ13C values of the motor oil samples
remained constant at -29%, while the fuel samples ranged from
-27.8 to -28.9%; we did not consider these results in our
calculations.
We determined the amount of biodiesel-derived carbon that
was accumulating in the used motor oils based on its 14C
content as follows. First, we apportioned the carbon of the
fuel blend with respect to the biodiesel component and motor
oil component using Δ14C mass balance
Δ14 C mixture ¼ Fc, bio Δ14 C bio þ ð1 - Fc, bio ÞΔ14 C motor ð1Þ
where Δ14Cmixture is the measured 14C content of used motor oil
via AMS. We assigned Δ14Cbio from the values of the B99.9
collected 3 times in the study. The Δ14Cmotor was fixed at a value
of -996% (on the basis of the measurement of the motor oil at
the beginning of the experiment). Finally, FC,bio is the mass
fraction of the total mixture of carbon that is derived from
biodiesel carbon. Rearranging eq 1, FC,bio can be expressed as
Δ14 C mixture - Δ14 C motor
FC, bio ¼ ð2Þ
Δ14 Cbio - Δ14 C motor
1042
Peacock et al.
Equation 2 shows that the proportion of biodiesel carbon in the
sample fuel blend (FC,bio) can be determined on the basis of the
measured Δ14Cmixture of the sample and the a priori known
Δ14Cbio and Δ14Cmotor values. The percentage of biodiesel
carbon is therefore
Δ14 C mixture -Δ14 C motor
%C, bio ¼
100 ð3Þ
Δ14 C bio - Δ14 C motor
Applying these calculations, we found that, after 591, 1556,
2210, and 3240 km, the %C,bio was 0.14 ( 0.04, 0.30 ( 0.05, 0.43
( 0.05, and 0.68 ( 0.06%, respectively. These results are similar
to the amount of FAMEs found in these samples with
GC-FID (Figure 2), indicating that the main source of biodie-
sel carbon is the FAMEs. Because we were not able to sample
every B99.9 used by this vehicle, it is possible that the Δ14C
values could have a wider range of values. In a previous study,
we found that B99.9 and B100 samples exhibited a Δ14C range
of þ10 ( 10%; therefore, we also applied these values in our
calculations.52 This did not change the results.
Conclusions
We investigated the accumulation of biodiesel components
in the motor oil of a biodiesel-operated vehicle. FAMEs
increased to 0.5% after the car had traveled 3240 km with
the same oil. We did not detect any PAHs in our motor oil
samples above our method detection limits for individual
compounds (5 μg g-1 of oil). In comparison, previous studies
with fossil-diesel- and gasoline-powered vehicles found total
PAH values of ∼20026,32 and 10 00026-31 μg g-1 of oil,
respectively. Radiocarbon analysis indicated that the FAMEs
measured with GC-FID were the only significant biodiesel
components in the motor oil samples.
Future work may employ this dual chromatographic and
isotopic approach for studies on different types of diesel
engines burning biodiesel, such as trucks, buses, or older,
more worn engines, as well as engines using different types of
motor oil. A parallel need also exists in the study of emissions
and source apportion PAHs from biodiesel vehicles,61 where
conflicting evidence has suggested either decrease, increase, or
no change in certain PAH emissions with the use of bio-
diesel.62,63 A complete understanding of PAH input from both
engine oil and emissions would allow for a true estimation of
the decreased input of these compounds to the environment
that would result from large-scale replacement of fossil diesel
with biodiesel. In addition, it would be prudent to subject
biodiesel engine oil to sensitive biological testing to further
ensure that it does not contain any mutagenic or carcinogenic
compounds at extremely low concentrations that would not
have been detected by this study. Overall, the current work
indicates that using biodiesel may decrease the input of PAHs
in the environment, relative to fossil diesel, related to the
leaking of motor oil from vehicles.
Acknowledgment. We thank Steven Volpin for providing the
samples for this study and Dr. Li Xu for assistance with the
radiocarbon analysis. We also thank Dr. John Farrington and
Dr. James Quinn for their thoughtful ideas and comments.
(61) Reddy, C. M.; Pearson, A.; Xu, L.; McNichol, A. P.; Benner,
B. A.; Wise, S. A.; Klouda, G.; Currie, L. A.; Eglinton, T. I. Environ. Sci.
Technol. 2002, 36, 1774–1782.
(62) Zou, L.; Atkinson, S. Environ. Technol. 2003, 24, 1253–1260.
(63) Karavalakis, G.; Alvanou, F.; Stournas, S.; Bakeas, E. Fuel 2009,
88, 1078–1085.