Characterization of Naphthenic Acids from

Athabasca Oil Sands Using Electrospray

Ionization: The Significant Influence of Solvents
John V. Headley* and Kerry M. Peru
Aquatic Ecosystem Protection Research Division, Water Science and Technology Directorate, Environment Canada,
11 Innovation Boulevard, Saskatoon, Saskatchewan, S7N 3H5, Canada
Mark P. Barrow and Peter J. Derrick
Institute of Mass Spectrometry and Department of Chemistry, University of Warwick, Coventry, CV4 7AL, United Kingdom
There is a need to develop routine and rugged methods
for the characterization of oil sands naphthenic acids
present in natural waters and contaminated soils. Mass
spectra of naphthenic acids, obtained using a variant of
electrospray ionization coupled with a Fourier transform
ion cyclotron resonance mass spectrometer, are shown
here to vary greatly, reflecting their dependence on
solubilities of the acids in organic solvents. The solubili-
ties of components in, for example, 1-octanol (similar
solvent to fatty tissue) compared to polar solvents such
as methanol or acetonitrile are used here as a surrogate
to indicate the more bioavailable or toxic components of
naphthenic acids in natural waters. Monocarboxylic com-
pounds (C
n
H
2n+z
O
2
) in the z ) -4, -6, and -12 (2-, 3-,
and 6-ring naphthenic acids, respectively) family in the
carbon number range of 13-19 were prevalent in all
solvent systems. The surrogate method is intended to
serve as a guide in the isolation of principle toxic com-
ponents, which in turn supports efforts to remediate oil
sands contaminated soils and groundwater.
Naphthenic Acids. There is a need to better characterize
naphthenic acids within crude oils and aquatic environments.
1,2
Naphthenic acids are defined as carboxylic acids that include one
or more saturated ring structures, though the definition has
become more loosely used to describe the range of organic acids
found within crude oil (Figure 1). The structural formulas for such
acids may be described by C
n
H
2n+z
O
2
,
3-7
where z is referred to
as the hydrogen deficiency and is a negative, even integer. More
than one isomer will exist for a given z homologue, and the
carboxylic acid group is usually bonded or attached to a side chain,
rather than directly to the cycloaliphatic ring.
3,4
The molecular
weights differ by 14 mass units (CH
2
) between n series and by
two mass units (H
2
) between z series.
8
Naphthenic acids can be
solubilized to produce metal salts (e.g., sodium and copper
naphthenates) that have industrial applications such as surfactants
and fungicides for wood preservatives.
6,9,10
Naphthenic Acids: The Oil Industry and Fate in the
Environment. Several studies have focused on the environmental
fate, transport, degradation, epidemiology, and isolation of specific
toxic naphthenic acids.
11
In a recent investigation of several
commercial naphthenic acid mixtures and those extracted from
oil sands, significant differences were established among four
commercial mixtures and between the extracts from various oil
sands ores and tailings ponds.
12
The concentrations and composi-
tion of the naphthenic acids were highly varied among commercial
sources, oil sands ore, and tailings pond sources.
12
The oil industry is concerned with the presence of naphthenic
acids in crude oils due to the association with corrosion. Crude
oil typically contain naphthenic acids in quantities of up to 4% by
weight, and characterization of the acids present within a sample
has become a topic of great interest due to the fact that the acids
corrode refinery units. Naphthenic acids are known to be weakly
biodegradable and therefore well-suited for use in identification
of oil source maturation.
13,14
Biodegradation of naphthenic acids
also occurs within oil reservoirs as the crude oil matures. Koike
* To whom correspondence should be addressed. E-mail: John.Headley@
ec.gc.ca.
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6222 Analytical Chemistry, Vol. 79, No. 16, August 15, 2007 10.1021/ac070905w CCC: $37.00 2007 American Chemical Society
Published on Web 06/30/2007
et al. have clearly demonstrated biodegradation of the low
molecular weight carboxylic acid fraction
15
in crude oil. The
susceptibility of crude oil to biodegradation has been shown to
increase following photolysis.
16-18
In unrefined Athabasca bitumen (northern Alberta, Canada),
the carboxylic fraction is 2%, of which 90% is composed of the
tricyclic acids that primarily make up the naphthenic acid
fraction.
19,20
The naphthenic acids may enter surface water systems
through such mechanisms as groundwater mixing and erosion
of riverbank oil deposits in oil-producing regions such as the
Athabasca oil sands.
21
The most significant possible environmental
receptor is suspected to be water due to direct contact with oil
sands material. Ambient levels in northern Alberta rivers in the
Athabasca oil sands are generally below 1 mg/L. However, tailings
pond waters may contain as much as 110 mg/L.
The relatively low aqueous solubility and moderately strong
sorption to soils are key factors limiting the bioavailability of oil
sands naphthenic acids in aquatic environments.
22
Naphthenic
acids are, however, known to be toxic to fish.
5
The aquatic toxicity
is associated with their concentration
23
and surfactant characteris-
tics.
24-27
There is little information about mammalian toxicity, but
the human lethal dosage was reported as 11 g kg
-128
and the oral
LD
50
for rats is between 3.0 and 5.2 g kg
-1
with death caused by
gastrointestinal disturbances.
29
Instrumental Techniques. Conventionally employed chro-
matographic methods are not effective for a complete separation
of the naphthenic acid species prior to mass spectrometric
detection. However, a variety of mass spectrometric techniques
have been used for the structural elucidation of naphthenic acids,
including useful approaches such as gas chromatography-mass
spectrometry,
3,6,16
liquid chromatography-mass spectrometry,
30
atmospheric pressure chemical ionization (APCI),
7
and electro-
spray ionization (ESI).
31-33
High mass accuracy and high resolution are a prerequisite for
full characterization of different ions of very similar mass, but same
class in complex mixtures. Although magnetic sector and FT-
ICR
34-36
instruments have been most commonly applied to such
investigations, time-of-flight mass spectrometry has also been used
to analyze crude oil samples.
37
Techniques such as APCI in
negative-ion mode can produce very clean spectra with good
sensitivity compared to other techniques.
7
Electrospray ionization,
however, is becoming the ionization technique of choice for the
mass spectrometry of naphthenic acids in crude oil samples.
Electrospray ionization
38-42
has the advantage of being an ioniza-
tion technique that imparts little energy to the nascent ions and
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Figure 1. Representative structures for oil sands naphthenic acids, showing alkylation sites. For z ) -4, for example, a typical range for n is
1-5. The average molecular weight of oil sands naphthenic acids in the Athabasca region is 260, with a preponderance of components in the
range of 180-300.
Analytical Chemistry, Vol. 79, No. 16, August 15, 2007 6223
results in the vaporization and ionization of a sample while
minimizing fragmentation. The combination of the ultrahigh mass
accuracy, ultrahigh resolution, and selective observation of the
deprotonated naphthenic acids make negative-ion mode FT-ICR
mass spectrometry an attractive technique for the characterization
of the naphthenic acids within a crude oil.
1,43-45
The ability to
determine the empirical formulas of the acidic species contained
within a given crude oil is of relevance to the continued fight
against corrosion within the petroleum industry and environmental
concerns.
2
Fourier transform ion cyclotron resonance mass spectra of
naphthenic acids are shown here to be strongly dependent on
the choice of solvents. The differences among the FT-ICR mass
spectra for different solvents are assumed to be primarily a result
of the selective solubilities of the respective components in a given
solvent. The electrospray process involves formation of colloidal
droplets, which shrink through evaporation of volatile solvents.
46
The more volatile the solvent, the more readily desolvation of
droplets occurs in the electrospray ionization process. The
consequence is effectively to concentrate the solution and to make
relevant the relative solubilities of the naphthenic acids in the
particular solvent. The solubilities of components in, for example,
1-octanol (similar solvent to fatty tissue) compared to polar
solvents such as methanol or acetonitrile are used here as a
surrogate to indicate the more bioavailable or toxic components
of naphthenic acids in natural waters. This approach is a simpli-
fication of the complex processes occurring in real world environ-
ments, as depending on the organism, the mode of living and
exposure (ingestion of benthos, dermal, across gills, etc.); the less
lipophilic, more polar fractions may also be important. However,
the surrogate method is intended to serve as a tool to guide
researchers in the isolation of principle toxic components, which
in turn supports efforts to remediate oil sands contaminated soils
and groundwater.
EXPERIMENTAL SECTION
The surrogate procedure for isolating potentially toxic and
bioavailable components of oil sands naphthenic acids in natural
waters partially depends upon the repeatability of the FT-ICR mass
spectra. Due care was given to experimental procedures to ensure
conditions were comparable from run to run. Concentration is
known to be an important factor, as there is mass spectrometric
evidence for naphthenic acids being prone to multimer formation.
47
The concentration at which multimer formation occurs is, how-
ever, strongly instrument dependent. In particular, adjustment of
parameters such as cone voltage and curtain gas can be used to
promote or eliminate multimers at a given concentration. Care
was therefore given to ensure that the same concentration and
instrument conditions were used in comparisons of respective
mass spectra. Changes in the mass spectra may also arise from
possible degradation of standards with time. To achieve the
optimum results, experiments were performed under the same
experimental conditions, as the temperature of the ions and fields
within the ion source are known to influence significantly the mass
spectrum.
48
Additionally, the same extract was used and the
samples were analyzed on the same day. The repeatability of the
FT-ICR mass spectra was determined using repeat, consecutive
runs (typically six) for the same solvent system under the same
instrumental conditions. For the octanol-acetonitrile solvent
system, runs with poor stability of the spray were discarded in
preference for runs acquired with a more stable spray. The
selection in this case was thus based upon the experience of the
instrument operator.
Instrumentation. Experiments were performed using a 9.4-T
Bruker BioApex II (Bruker Daltonics, Billerica, MA) Fourier
transform ion cyclotron resonance mass spectrometer.
49
The
superconducting magnet (Magnex Ltd.) was maintained at the
operating temperature using liquid helium and liquid nitrogen,
where the entirety of the magnet was enclosed by passive
shielding. The heart of the instrument was the Infinity Cell,
50
which was cylindrical in geometry, rather than the traditional
cubic, and consisted of six plates: two trapping plates, two
detection plates, and two excitation plates. Segmentation of the
trapping plates, and the application of differing radio frequency
(rf) potentials to these segments, lead to homogeneous electro-
static fields from the excitation plates that simulates an infinitely
long cell. The advantage of the cell design was the minimization
of ion loss during excitation, which can arise from the effect of
radial components of the excitation field; such ion loss can lead
to discrimination effects during the excitation stage of an experi-
ment. During the course of these experiments, a trapping potential
(PV1 and PV2) of -1.5 V was maintained to constrain the ions
movement within the cell and detection was performed over a
range of m/z 144-3000, with the excitation range being slightly
wider to compensate for possible inhomogeneities at the edges
of the rf chirp used for excitation.
The flight tube was pumped by three cryogenic pumps, which
maintained a base pressure of the order of 10
-10
mbar within the
cell region during experiments. A base pressure of the order of
10
-7
mbar was maintained within the interface between an external
ion source and the flight tube. Ions were accumulated within the
hexapole ion trap for a period of 2 s (D1 ) 2 s) prior to extraction.
For a user-defined period of time, ions were extracted from the
hexapole, traversed the ion optics, and entered the FT-ICR cell.
The period of extraction of ions from the hexapole, referred to as
P2, was set to 2400 s for the course of these experiments. The
SideKick mechanism was employed to deflect ions off of the ion
optical axis as they enter the cell and increase trapping efficiency
(EV1 ) 1.09 V, EV2 ) 1.60 V; where EV values corresponded to
ICR cell extraction plates), DEV2 (ICR cell delta extraction plate
2) ) -4.13 V). Dipolar excitation was used to excite the ions to
(42) Mann, M., Ed. Electrospray Mass Spectrometry; Kluwer Academic Publish-
ers: Dordrecht, The Netherlands, 1992.
(43) Qian, K.; Robbins, W. K.; Hughey, C. A.; Cooper, H. J.; Rodgers, R. P.;
Marshall, A. G. Energy Fuels 2001, 15, 1505-1511.
(44) Brandal, O.; Hanneseth, A. M.; Hemmingsen, P. V.; Sjoblom, J.; Kim, S.;
Rodgers, R. P.; Marshall, A. G. J. Dispersion Sci. Technol. 2006, 27, 295-
305.
(45) Hemmingsen, P. V.; Kim, S.; Pettersen, H. E.; Rodgers, R. P.; Sjoblom, J.;
Marshall, A. G. Energy Fuels 2006, 20, 1980-1987.
(46) Peschke, M.; Verkerk, U. H.; Kebarle, P. J. Am. Soc. Mass Spectrom. 2004,
15, 1424-1434.
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2002, 85, 182-187.
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1812-1822.
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2000, 6, 267-275.
(50) Caravatti, P.; Allemann, M. Org. Mass Spectrom. 1991, 26, 514-518.
6224 Analytical Chemistry, Vol. 79, No. 16, August 15, 2007
a detectable cyclotron orbit prior to the detection stage. The
duration for the excitation steps, P3, was set to 12 s, the rf
attenuation for the excitation, PL3, was set to 10.15, and XBB,
which determines the excitation step size, was maintained at a
value of 200. Data acquisition in broadband mode used a 12-bit,
10-MHz digitizer. This maximum sampling rate can be translated
to a minimum mass-to-charge ratio of 29 on the basis of the
Nyquist criterion, which states that the sampling frequency must
be at least equal to twice the frequency being sampled. The
instrument was controlled using a Silicon Graphics Indy worksta-
tion running XMASS 5.0.10 (Bruker Daltonics, Billerica, MA)
under IRIX. 5.3. Data files consisted of 512 K (524 288) data points.
The raw data were electronically converted from the time domain
to the frequency domain via a fast Fourier transform.
Oil Sands Naphthenic Acids Samples. The samples of
interest were obtained from four sites at an industrial location in
the oil sands region of the Athabasca River Basin, Alberta, Canada.
The naphthenic acid concentrate was obtained by extraction of
oil sands process water following procedures described by Rogers
et al.
26
The extracted naphthenic acid concentrate was made up
in acetonitrile to a measured concentration of 8500 mg/L.
This naphthenic acid extract was used to prepare soluble
fractions in five separate solvents using a ratio of 1 mg of extract
to 1 mL of solvent. Visible differences in solubility were evident
particularly for the 1-octanol/acetonitrile solvent system. The
resulting solvent systems, with the polarity indices given in
parentheses for the solvents, were as follows: Milli-Q water (9.0)/
acetonitrile (5.8); acetonitrile (5.8); methanol (5.1)/acetonitrile
(5.8); dichloromethane (3.1)/acetonitrile (5.8); and 1-octanol
(<3.9)/acetonitrile (5.8). An aliquot of 1% (by volume) ammonia
solution (35% by volume in water) was added to the solutions to
assist deprotonation of the naphthenic acid species.
Ion Source Conditions. Nanospray was chosen instead of
normal ESI. The latter gave comparable results with a steady spray
for the octanol/acetonitrile cosolvent, but gave very poor desol-
vation in the ion source region for this cosolvent and required
considerably more sample for a given experiment. The nanospray
apparatus was based on an electrospray ion source (Analytica of
Branford), with a metal-coated glass needle replacing the arrange-
ment of a metal needle connected in series to a syringe pump.
Sample solution (10-20 L) was placed in the metal-coated glass
needle (nanospray needle), and the needle was secured within a
mount. These nanospray needles had been purchased from
Proxeon Biosystems (formerly Protana, Odense, Denmark). A gas
line was connected to the mount to provide a propelling gas, with
carbon dioxide being the gas of choice at a pressure typically of
10 psi. The nanospray needles had sealed tips, which needed to
be broken prior to initialization of the spray. Using a microscope
to view the process, the sealed needle tip was broken carefully
on the protective end cap placed over the Pyrex capillary, which
allowed the initiation of the spray of the sample solution. The
capillary-skimmer region of an electrospray ion source plays an
important role,
48
and potential at the inside end of the capillary
was maintained at -110.40 V and that on the skimmer at -4.00
V in all experiments. The potential difference between the nano-
spray needle and the outside end of the capillary was namely
300 V.
Data Processing. ESI Tuning Mix (Agilent, Palo Alto, CA)
was used to tune and calibrate externally the instrument.
The calibration was compared with that obtained through
usage of a Kodak naphthenic acid mixture (Kodak Chemicals,
Rochester, NY), and the two, separately obtained, external
calibrations were virtually indistinguishable when applied
to the data. As illustrated in Figure 2, over 70% of the signals
(m/z 199-379) had a mass accuracy of 0.5 ppm or better, and
86% of the signals had an associated mass error of 1 ppm or
less. The combination of high mass accuracy and high reso-
lution, allowing closely spaced signals within a complex
mixture to be distinguished, makes FT-ICR mass spectro-
metry a preferred technique for the analysis of com-
Figure 2. Representative mass accuracies obtained for oil sands naphthenic acids, illustrated using data for the Milli-Q water/acetonitrile
solvent system. The mass error was typically less than 0.5 ppm for most assignments (over 70%).
Analytical Chemistry, Vol. 79, No. 16, August 15, 2007 6225
plex mixtures, where assignment of empirical formulas is
important.
Once the mass spectra were obtained, the data were processed
using XMASS. The relevant signals were categorized according
to their z number, or hydrogen deficiency, using a custom-
designed Microsoft Excel spreadsheet. The sorted data were
transferred to Microcal Origin prior to the creation of the plots
of the naphthenic acid content of each sample. The results were
displayed as profiles of the relative intensities of the signals as a
function of the carbon content (n), for the different hydrogen
deficiencies. Such plots can serve as fingerprints for different
samples, when obtained under similar experimental conditions.
Conversely, it is possible to study a single sample and monitor
how the profiles are affected by experimental conditions, as
described here.
RESULTS AND DISCUSSION
The efficiency of the spray in electrospray ionization is
dependent on the physical properties of the solvent. In this work,
it was relatively easy to obtain a reliable nanospray signal (as
evidenced by measuring the ion current and monitoring the mass
spectra on a scan-by-scan basis) for all naphthenic acid extracts
investigated when using acetonitrile alone as the solvent, but this
was not possible using 1-octanol alone or 1-octanol/water solvents.
Acetonitrile was thus used as a cosolvent in all the solvent
mixtures investigated to avoid irreproducibility arising from
instability during spraying. The nature of the spray in the
electrospray ionization is influenced by factors including surface
free energy, the viscosity of the solvent, size of droplets, and
desolvation of droplets. The appearance of the FT-ICR mass
spectra will depend upon the spray conditions. For the purposes
of the surrogate method evaluated herein, differences in FT-ICR
mass spectra are assumed to be predominantly due to the choice
of solvent, because all other ion source conditions were kept
constant.
Headley et al. reported that the negative-ion ESI mass spectra
of Fluka naphthenic acids spiked in natural waters were strongly
dependent on the pH of the water.
47
The mass spectra acquired
for the Fluka naphthenic acid mixture were less complex when
obtained at lower pH, compared to pHs greater than 7. It will be
demonstrated here that the FT-ICR mass spectra of the oil sands
naphthenic acids are also dependent on the choice of solvent
system.
In general, the FT-ICR mass spectra for many of the solvent
systems investigated were very similar, but key differences were
evident for the 1-octanol/acetonitrile and dichloromethane/
acetonitrile cosolvents. The majority of solvent systems were
characterized by a number-average molecular weight (N
i
M
i
/
N
i
) of 260, with a preponderance of naphthenic acids in the range
of m/z 180-300. The major difference observed for the 1-oc-
tanol/acetonitrile cosolvent was the appearance of a wider range
of naphthenic acids at heavier molecular weights in the range of
m/z 300-370. The latter are presumably less polar but selectively
enriched for detection in the less polar 1-octanol/acetonitrile
cosolvent, compared to the other cosolvents investigated. The key
difference for the dichloromethane/acetonitrile cosolvent was the
low abundance of C
n
H
2n+z
O
4
species, discussion of which is given
later. For the purpose of clarity when illustrating the surrogate
method presented here, the discussion has been limited to
C
n
H
2n+z
O
2
and C
n
H
2n+z
O
4
species and will not extend to hetero-
atomic species containing N, S, or O
3
species. Full evaluation of
the latter and other species is deferred to a future communication.
Figure 3. Negative-ion mode broadband mass spectra of a sample from a site in the oil sands region of the Athabasca River basin, Alberta,
Canada. The sample solutions were based upon usage of (a) acetonitrile alone, (b) methanol/acetonitrile (1:1), or (c) Milli-Q water/acetonitrile
(1:1).
6226 Analytical Chemistry, Vol. 79, No. 16, August 15, 2007
The similarity of the FT-ICR mass spectra using the acetonitrile
(only), methanol/acetonitrile (1:1), and Milli-Q water/acetonitrile
(1:1) solvent systems is evident from Figure 3 for the naphthenic
acid extract. The m/z ranges are similar for the respective
solvents, with minor differences within the m/z 200-250 region.
For most runs, the distribution of molecular weights was similar
with a number-average molecular weight of 260. These polar
solvents were apparently well suited for the polar fraction of the
oil sands naphthenic acids.
Figure 4 shows the FT-ICR mass spectra obtained using
three solvent systems: (a) dichloromethane/acetonitrile (1:1),
(b) water/acetonitrile (1:1), and (c) 1-octanol/acetonitrile (1:1).
The mass spectrum shown in Figure 4 b is representative
of the three mass spectra shown in Figure 3, acetonitrile only,
methanol/acetonitrile (1:1), and water/acetonitrile (1:1). The
mass spectrum obtained using dichloromethane/acetonitrile ex-
hibits a smaller number of peaks, while the mass spectrum
acquired using 1-octanol/acetonitrile displays a larger number of
peaks.
The FT-ICR data have been processed and replotted to show
the respective compounds of a given z series in the mixture. The
presentation of the data as such illustrates how observation of
each naphthenic acid z series is dependent on the solvent system.
The respective plots for the water/acetonitrile, Milli-Q water/
acetonitrile, and the 1-octanol/acetonitrile cosolvents are given
in Figure 5. The C
n
H
2n+z
O
2
series are shown in Figure 5a, c, and
e, while the C
n
H
2n+z
O
4
series are shown in Figure 5b, d, and f.
The monocarboxylic compounds in the z ) -4, -6, and -12
families in the carbon number range of 13-19 are among the most
prevalent for all three solvent systems. These components along
with other naphthenic acids in the range z ) 0 to z ) -12 have
been reported to sorb strongly to soils in aquatic systems. Their
removal from the water column by sorption is particularly strong
in the presence of salts found in groundwater.
51
Taking the results
obtained with the water/acetonitrile solvent system as a bench-
mark, the results acquired using 1-octanol/acetonitrile and dichlo-
romethane/acetonitrile can be compared. The C
n
H
2n+z
O
2
species
are relatively less intense in the 1-octanol/acetonitrile solvent
system (comparing Figure 5e and c), while, conversely, it is the
C
n
H
2n+z
O
4
species that are relatively less intense when using the
dichloromethane/acetonitrile solvent system (comparing Figure
5b and d).
The solubility of some components with molecular weights
above 300 amu in 1-octanol/acetonitrile cosolvent compared to
the other solvents investigated is apparently higher. There
appears, to be selective enrichment of the z ) -6 series for the
dicarboxylic or C
n
H
2n+z
O
4
naphthenic acids, as illustrated in Figure
5f. The reasons for this apparent enrichment are not established
and may be due in part to differences in polarity and structure of
the naphthenic acids. Furthermore, it is evident that a key factor
controlling the solubility in the 1-octanol/acetonitrile solvent
system is the size or molecular weight of the components. As the
molecular weight increases, the influence of the hydrocarbon tail
of the naphthenic acid may become less pronounced for selective
solubility in 1-octanol. By contrast, the distribution of the C
n
H
2n+z
O
4
species were low to nondetectable in the dichloromethane/
acetonitrile cosolvent, as seen in Figure 5b. The z ) -6 and z )
-8 families are among the most visible, in a manner similar to
profiles seen in Figure 5d (water/acetonitrile) and Figure 5f (1-
octanol/acetonitrile). The low abundance of the C
n
H
2n+z
O
4
species appears to be limited to the dichloromethane/acetoni-
(51) Janfada, A.; Headley, J. V.; Peru, K. M.; Barbour, S. L. J. Environ. Sci. Health
A 2006, 41, 985-997.
Figure 4. Comparison of the mass spectra acquired using (a) dichloromethane/acetonitrile (1:1), (b) Milli-Q water/acetonitrile (1:1), or (c)
1-octanol/acetonitrile (1:1) solvent systems.
Analytical Chemistry, Vol. 79, No. 16, August 15, 2007 6227
trile system and was not observed for the other cosolvents
investigated.
These findings have implications for the toxicity and trans-
port of naphthenic acids in water since the more soluble
components in 1-octanol are expected to be more bioavailable for
ingestion and less persistent in tailings, oil sands sludges, and
groundwater. The pronounced trace for the C
n
H
2n-6
O
4
species in
1-octanol/acetonitrile, relative to the other solvents, may thus be
indicative of their bioavailability according to the surrogate
method put forward here. The dicarboxylic components with
carbon number in the C
11
-C
16
range can be prevalent in Atha-
basca oil sands. Given that such components are likely to be
readily biodegradable, their removal from aquatic systems is
expected to alter the distribution of the molecular weights
of the naphthenic acid mixture significantly. This trend is
consistent with results reported for shifts in the average mole-
cular weight distribution of naphthenic acids observed for
fresh and aged oil sands tailing pond waters. As the more
prevalent, lower molecular weight, bioavailable, and acutely
toxic fractions of naphthenic acids are biodegraded over
time, the spectra of the molecular weight distribution shift to
higher molecular weight recalcitrant components in aged
materials.
10,23,25,52
As noted earlier, there was difficulty using 1-octanol as a solvent
in the nanospray experiments. The use of the cosolvent 1-octanol/
acetonitrile resulted in improvements but was only suc-
cessful for extracts from one of four sites studied in the
Athabasca oil sands region. This finding is probably ascribed
to the presence of a matrix effect in the investigated samples.
Further work is thus warranted to determine under what
conditions 1-octanol and possibly other solvents, such as di-
myristoylphosphatidylcholine (a synthetic lipid, commonly
known as DMPC), may be used in conjunction with electro-
spray ionization to mimic bioavailability of naphthenic
acids.
It has been demonstrated that, while there was little change
in the FT-ICR mass spectra for some of the polar solvents
investigated, there were in contrast dramatic effects in the
appearance of the mass spectra observed for the 1-octanol/
acetonitrile and dichloromethane/acetonitrile solvent sys-
(52) Clemente, J. S.; Yen, T.-W.; Fedorak, P. M. J. Environ. Eng. Sci. 2003, 2,
177-189.
Figure 5. FT-ICR MS data showing naphthenic acid z series dependence on the different solvent systems for dichloromethane/acetonitrile,
Milli-Q water/acetonitrile, and 1-octanol/acetonitrile cosolvents. For the purpose of simplicity, the CnH2n+zO2 series are shown in (a), (c), and (e),
while the CnH2n+zO4 series are shown in (b), (d), and (f).
6228 Analytical Chemistry, Vol. 79, No. 16, August 15, 2007
tems. It is hypothesized that this difference reflects the differ-
ent solubilities of the various naphthenic acids in these
solvents, and that in turn, this variability reflects their preferen-
tial availability for uptake by biota in aquatic systems. Re-
search is continuing in support of remediation efforts to de-
velop and test the aquatic toxicity of the naphthenic acids
fractionated according to their respective solubilities in organic
solvents.
ACKNOWLEDGMENT
This work was funded in part by the Program of Energy
Research and Development.
Received for review May 2, 2007. Accepted May 30, 2007.
AC070905W
Analytical Chemistry, Vol. 79, No. 16, August 15, 2007 6229