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An Improved Gravimetric Method to Determine Total Petroleum

Hydrocarbons in Contaminated Soils

Article  in  Water Air and Soil Pollution · October 2008

DOI: 10.1007/s11270-008-9704-1

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Water Air Soil Pollut (2008) 194:151–161
DOI 10.1007/s11270-008-9704-1

An Improved Gravimetric Method to Determine Total

Petroleum Hydrocarbons in Contaminated Soils
Mario Villalobos & Ana Paulina Avila-Forcada &
Margarita Eugenia Gutierrez-Ruiz

Received: 26 November 2007 / Accepted: 28 March 2008 / Published online: 25 April 2008
# Springer Science + Business Media B.V. 2008

Abstract A gravimetric method to determine heavy
fractions of total petroleum hydrocarbons (TPH) in
soils is reported. The method was adapted and
calibrated by modifying previous standard methods
published, incorporating energy and cost savings
where possible. Artificially contaminated soils with
different organic matter content, and aged in stationary
mode for a period of 8 months were used for
calibration. Insufficient solvent evaporation was iden-
tified as the most prevalent and largest positive
interference in the gravimetric detection. To overcome
this, while minimizing the need for heating, a combi-
nation of three 10-min rotary evaporator steps and
30 min of vacuum in a desiccator were applied, for a
total solvent volume of 60 ml. Hexane was chosen as
the extraction solvent and a 40–60 min treatment in an
ultrasound bath of 260 W was found suitable to extract
80–95% of TPH extracted by the Soxhlet method.
Finally, the use of silica gel for cleanup of co-extracted
natural organic matter was found unnecessary, because
of the low amounts co-extracted for soils with up to 5%
organic carbon, and because the chemical nature of the
co-extracted organic matter prevents its selective
adsorption to silica.
Keywords Contaminated soils . Gravimetric method .
Silica gel . Solvent evaporation . Total petroleum
hydrocarbons . Ultrasound extraction

1 Introduction
M. Villalobos (*) : M. E. Gutierrez-Ruiz
Grupo de Bio-Geoquímica Ambiental,
Laboratorio de FisicoQuímica Ambiental (LAFQA)
Soil pollution by petroleum hydrocarbons usually
Instituto de Geografía, originates from spills or leaks of storage tanks during
Universidad Nacional Autónoma de México (UNAM), fuel supply and discharge operations. Most petroleum
Coyoacán 04510 D.F., México components are hazardous to human health and to the
e-mail: marvilla@igg.unam.mx
soil biota (MDEP 1994); therefore, measurement of their
M. E. Gutierrez-Ruiz total concentration (total petroleum hydrocarbons—
e-mail: ginny@servidor.unam.mx
TPH) is the initial and most general indicator of hazard
A. P. Avila-Forcada by hydrocarbons for environmental diagnostic purposes.
Dirección General del Centro Nacional de Investigación y The US Environmental Protection Agency (USEPA)
Capacitación Ambiental (CENICA), has published a series of methods for quantifying TPH
Instituto Nacional de Ecología, San Rafael Atlixco 186,
in solid matrices (including soils; Table 1). The four
Col. Vicentina,
Iztapalapa. D.F. 09340, México, México categories of analytical methods described in Table 1
e-mail: apavila@ine.gob.mx are based on the final detection method, i.e. gravimetry,
152 Water Air Soil Pollut (2008) 194:151–161

Table 1 USEPA methods to quantify TPH in various matrices (including soil; taken from Weisman, 1998)

Detection EPA method Extracting solvent and Approximate Aprox. Cost/ Advantages Disadvantages
method no. (year of most range of hydrocarbon detection limit, samplea
recent revision) species extracted mg/kg (soil)

Gravimetric 9071 B (1998) n-Hexane. Most 50 50 Simple, quick, Lack of specificity, low
1664 A, for compounds except and inexpensive sensibility, high loss of
water samples volatile fractions volatiles, prone to
(1999) interferences; only
useful for total
quantification.
Infrared (IR) 8440 (1996) Supercritical CO2. 10 50–80 Simple, quick, Lack of specificity, low
Most compounds and inexpensive sensitivity, high loss of
except volatile and volatiles, poor
very heavy fractions extraction of high
molecular weight
hydrocarbons, prone to
interferences; only
useful for total
quantification
Gas chromatography 8015 C (2000) Methanol, n-hexane, 10 100–150 Detects a broad Normally it cannot detect
(GC) 2-propanol, range of compounds below C6;
ciclohexane, hydrocarbons does not detect polar
acetonitrile, acetone, compounds; hydrocarbons (alcohols,
methylene chloride, or simple and ethers, etc.); chlorinated
mixtures of these. sensitive; solvents may be
Normally between C6 chemical quantified as TPH
and C25 or C36 (may speciation is
be modified for a higher possible.
range)
Immunoassay 4030 (1996) The extractant depends 10–500 13b Simple, quick, Low sensitivity, prone to
on the commercial kit. 64c inexpensive, interferences, primarily
Aromatic hydrocarbons and may be measures aromatics; low
(BTEX, PAHs) used in the field accuracy and precision;
must be used as a
screening method; only
useful for total
quantification
a
US dollars, MDEP (1994)
b
CMECC (1996)
c
VRAI (2007)

infrared spectroscopy, gas chromatography, and immu-
noassays. Currently, Mexican legislation has imple-
mented many of these as standard methods for
diagnosis of contaminated soil sites, especially meth-
ods that employ various forms of gas chromatographic
techniques, but also a gravimetric method has been
recently adopted (NMX-AA-134-SCFI-2006 2006).
Chronologically, gravimetric methods were among
the first ones developed, but were readily superseded
with the advent of more specific spectroscopic and
chromatographic methods that additionally allow for
varying degrees of hydrocarbon chemical speciation.
Presumably, for this reason, gravimetric methods
show insufficient investigation pertaining proper
method development, calibration, and validation that
allow identification of the important variables that
control interferences to the final gravimetric measure-
ment. For example, it is not clear what conditions are
required (and what their sensitivity is) to ensure
complete evaporation of the solvent once the TPH
extraction has been performed. Frequently, the proce-
dures are vaguely stated to complete distillation in
less than 30 minutes (EPA Method 9071 B 1998;
NMX-AA-134-SCFI-2006), but no indication is given
Water Air Soil Pollut (2008) 194:151–161
about the way to guarantee or to quantify such
complete distillation. Thus, there is a high potential
either for TPH overestimation, or for over expenditure
of energy to ensure complete distillation, which
increases the costs of analyses.
For laboratories with low resources, such as rural
laboratories in developing countries, gravimetry is the
method of choice due to the lower costs involved as
compared to other methods. The most basic laboratory
is equipped with an analytical balance as a basic tool,
and thus, gravimetric methods require no additional
expense for detection of hydrocarbons. Additionally,
gravimetry allows for simplicity and low-cost analysis
per sample (Table 1), which decreases considerably
overall costs in situations where large numbers of
samples are required for analysis, as may occur in
conditions of high soil pollution heterogeneity.
Gravimetric detection is non-specific, although
quite sensitive, and the potential for interferences is
high. The present work focuses on developing/
calibrating a gravimetric method for non-volatile
TPH quantification in contaminated soils, in which
cost-savings, simplicity, and speed have been defined
as sought-after attributes for general applicability in
low-resources laboratories. This investigation provided
the procedures to recognize the important parameters for
accurate quantification of TPH, identifying especially
the highly sensitive steps that control the final precision
and accuracy of the method.
The starting gravimetric method chosen in the
present work is EPA Method 9071 B (1998), in
combination with extraction EPA Method 3550 B
(1996), which utilizes ultrasound treatment, both of
which form the basis for the official Mexican norm
NMX-AA-134-SCFI-2006. Ultrasound was chosen
over Soxhlet extraction to further decrease costs.
The latter method provides a highly quantitative
extraction of TPH but is considerably more costly
time- and energy-wise. Furthermore, as a variation
from the published methods, the use of an ultrasound
water bath instead of a probe horn was investigated,
due to the higher availability of the former as basic
lab equipment. Three principal factors were investi-
gated in the overall method proposed: Extraction
efficiency, quantitative evaporation of the extracting
solvent, and co-extraction of natural organic matter. In
this latter, the need for silica-gel cleanup of the
extract, as recommended in some reference methods,
was additionally investigated.
153
2 Materials and Methods
2.1 General Gravimetric Method
Soil samples were sieved (Sieve 10, ASTM—2 mm,
followed by Sieve 200, ASTM—75 μm), and dried to
105°C for 12 h (Schwab et al. 1999; Xie et al. 1999).
These were homogenized mechanically for several
hours, and approximate accurately weighed 10 g
subsamples were placed in round flasks previously
dried (105°C) to constant weight. Ten grams of
anhydrous Na2SO4 were added to create a free-
flowing powder (EPA Method 3630 B 1996), and
35 ml n-hexane (here to forth referred to simply as
hexane) were used for extraction in an ultrasound bath
(EPA Method 3550 B 1996), the conditions of which
were investigated for optimal extraction efficiency (cf.
Section 3). The extracts were filtered through a
column packed first with 0.6 g treated cotton or glass
fibre, and optionally second and third with 5 g treated
silica gel, and 1 g treated celite (EPA Method 3630 B
1996), respectively, and washed down with additional
25 ml hexane to complete 60 ml in the final liquid
extract for analysis. The hexane was evaporated in a
rotary evaporator, followed by drying of the flask
outer walls with lint-free absorbent paper and evap-
oration of the remnant hexane under variable con-
ditions (cf. Section 3). The residue was weighed in an
analytical balance and designated as TPH.
The equipment and material used were: a semi-
microanalytical Sartorious R 2000 balance; a Heidolph
Laborota Digital 4003 rotary evaporator, equipped with
Rotovac Senso vaccum system, and Rotacool cooling
system (the conditions on the rotary evaporator were as
follows: Water bath temperature=40°C, Cooling water
temperature=14°C, Rotation speed=70 rpm, automatic
mode. Subsequently the flasks were dried with absor-
bent paper and placed in a desiccator for 2 h); two
ultrasonic baths of 5.7 l: a Bransonic, Branson 3510
R-MTH of 100 watts, and a Fisher Scientific, FS60, of
260 watts; two Pyrex glass desiccators with silica gel
desiccant: one of 7.5 l with plate diameter of 230 mm,
and a second one of 2.2 l and plate diameter 140 mm
connected to lab vacuum; a Felisa 243, Series 87013
oven of 1500 W; 250 ml Erlenmeyer Pyrex flasks; 50 ml
flat-bottom round Pyrex flasks; and glass columns of
250 mm length×10 mm internal diameter.
The reagents employed were Baker analytical
reagents (n-hexane, acetone, silica gel 40–140, and
154
anhydrous sodium sulfate). Celite (Química Meyer)
and cotton (Protec) were industrial grade. The diesel
used was from PEMEX (Mexican State Petroleum
Company), and a Diesel/Lubricating Oil Standard
(BAM 2000) was utilized. Hexane was chosen as the
extractions solvent due to its lower toxicity, in
contrast to other solvents, such as dichloromethane,
which have been identified previously as ideal in
mechanical shaking extractions (Schwab et al. 1999).
Cotton Treatment Washed first with acetone, and then
with hexane both in ultrasound bath for 25 min. Each
solvent was decanted and the remnant evaporated in
the hood after the washing procedure (EPA Method
3550 B 1996).
Silica Gel Treatment Washed first with hexane in
ultrasound bath for 25 min. Filtered, and washed
again with fresh hexane, which was decanted. Left in
the hood all night to ensure dryness. ‘Activation’ was
performed at 150°C for at least 16 h (EPA Method
3630 B 1996).
Celite Treatment Washed with distilled water in an
ultrasound bath for 25 min, and filtered. Washed
secondly with dicholoromethane for 25 min in
ultrasound bath. Filtered and dried in hood all night.
Heated in oven to 150°C for 4 h (Ricardo Alfaro,
Chemistry Institute, UNAM, personal communication).
Soxhlet Extraction Ten grams of soil samples were
accurately weighed in duplicate in Whatman thimbles,
and placed inside the glass tube of a Soxhlet apparatus
with 180 ml hexane extractant. An empty thimble as
blank was also processed. The extraction was per-
formed for 8 h to ensure maximum recovery, after
which the extract was concentrated to 40-ml total
Table 2 Main characteristics of non-contaminated soils used in the preparation of TPH-contaminated soil standards
Code Soil descriptor pH
F Fertilized 7.7
S Sodic saline 9.5
V Vertisol 7.1
R Red 7.2
Soils provided by courtesy of Dr. Arturo Aguirre Gómez, Department of Chemistry, FES Cuautitlán, UNAM.
a
Analysed in triplicate according to Aguirre et al. (2002)
Water Air Soil Pollut (2008) 194:151–161
volume in the rotary evaporator and filtered through a
cotton-packed column, washing with additional hex-
ane to make a total volume of 60 ml. The procedure
described in the general method was followed from this
point onwards.
2.2 Preparation of Standard Soils Contaminated
with TPH
Four non-contaminated soils were characterised (cf.
Table 2) and artificially contaminated and aged in a
stationary mode for 8 months, to be used in the final
calibration of the method, especially pertaining
optimization of the extraction procedure. Diesel was
chosen as representative of relatively heavy TPH
fractions, due to its homogeneous composition (well
balanced between aliphatic and aromatic compounds
in a C9 to C28 range; Schwab et al. 1999), and high
boiling point (about 400°C).
For the artificial contamination (BAM 2002; Arce
2000), soils were sieved and dried as described above,
and 100 g lots of each were weighed in round flasks
of 1,000 ml. Mixtures of different amounts of diesel,
accurately weighed, and dissolved in approximately
100 ml hexane were added to each lot, and homog-
enized by shaking for 6 h, followed by evaporation of
the bulk hexane in the rotary evaporator. Soils were
left overnight inside the hood (to achieve remnant
hexane evaporation) covered with aluminium foil, and
then kept stationary in the lab for 8 months. This
provided conditions for diesel sorption to the solid
minerals and natural organic matter (NOM) that were
more representative of aged contamination scenarios,
as compared to fresh contact. For each soil, five
samples were contaminated with the diesel concen-
trations shown below, which represent a wide range
Electric conductivity (mS/cm) % Organic carbon (OC)a
0.730 1.72
1.322 0.46
1.500 4.56
1.224 0.57
Water Air Soil Pollut (2008) 194:151–161
of TPH soil contamination levels (from 0.1% to 10%
in weight):
Soil number code 1 2 3 4
Applied diesel concentration 1,018 2,135 20,103 100,110 0
(mg/kg)
In the following section the details of the proce-
dures carried out and the parameters studied to
calibrate the method are described in the order in
which they were performed, providing proper sensi-
tivity tests to these parameters.
2.3 Detection Limit (DL) and Quantification Limit
(QL)
The DL definition adopted here is the one obtained
from three standard deviations of the blank. In this
particular case, because no standard soil with non-
hexane-extractable NOM content was available that
would serve as adequate blank, we limit this to a
calculation of a theoretical DL, taken from the
uncertainty of the analytical balance of 0.0005 g.
The DL is thus 0.0015 g. If this figure comes from a
10 g soil sample processed by the method, the overall
DL corresponds to 150 mg/kg. The quantification
limit, defined as 10 times the uncertainty of the
balance, i.e. 5 mg, translates to an overall QL of
500 mg/kg for a 10 g soil sample extraction. Analysis
of detection limits will be discussed below for soils
with hexane-extractable NOM.
2.4 Chromatographic Detection of Hexane-Extracted
Soil NOM and TPH Standards
Gas chromatograms were generated from the soil
NOM extracted by hexane, using GC-FID (Varian
Star 3400 CX series) and compared to those generated
under the same conditions for two TPH standards
(commercial PEMEX Diesel used in the present work,
and certified diesel/lubricating oil BAM standard
mixture; BAM 2002), and for the extracts of an
actual petroleum+oil−contaminated soil and several
non-contaminated soils, including one adjacent to the
contaminated one. The GC conditions applied for
these experiments were: Column injection technique,
DB-5 (30 m × 0.25 mm ID × 0.25 μm) column;
nitrogen carrier gas (1 ml/min); flame ionization
detector temperature, 320°C; furnace program, 70°C
(isotherm, 0.5 min) to 125°C (rate, 8°C/min) to 245°C
155
(rate, 6°C/min) to 300°C (rate, 4°C/min); total time,
49 min.
5
3 Results and Discussion
3.1 Evaporation of the Hexane Extractant
In gravimetric methods any mass that contributes to
the final measured extract weight not originating from
the analyte of interest generates a positive interference.
These types of interferences are the most common ones
in such methods when applied to measurements of
heavy-fraction TPH, and it’s of utmost importance to
minimize them. Principally, two steps in the procedure
for evaporating the extractant potentially contribute to
such positive interferences: (1) water sorbed to the
glass of flasks placed on the rotary evaporator, and
especially (2) insufficient evaporation of the solvent
used, because of its high relative initial volume.
The mass contributed by (1) was found to be
negligible for the glass flasks utilized, either from
drying at 105°C flasks that were not previously oven-
dried (error<0.0007 g), or from previously oven-dried
flasks at 105°C that were subjected to the conditions
of the rotary evaporator procedure, simulating the
solvent evaporation step, and placed in the desiccator
for 2 h after drying externally with lint-free absorbent
paper, (error<0.0005 g).
Optimal conditions to achieve quantitative hexane
evaporation were investigated in sets of duplicate
flasks to which approximate amounts of 0.6 g diesel
were accurately weighed in each, corresponding to
hypothetical 60,000 mg/kg TPH extracted from 10 g
of soil, and were mixed with 60 ml hexane. Two
rotary evaporator time length regimes were investi-
gated: (1) varying a first time step from 15 to 120 min
while maintaining a second step constant at 10 min;
and (2) applying three 10-min time-steps to achieve
higher evaporation efficiencies.
Figure 1 shows the average results obtained for the
set of experiments (1). Evaporation times above
60 min were necessary to achieve quantitative hexane
evaporation, but losses of diesel were observed at
higher times, producing negative errors.
The practical application of this experiment to real
samples has two limitations. One arises from the high
sensitivity of the final evaporated mass to the
evaporation time, in such a way that a very accurate
156
0.2500
Final hexane weight/(g)
0.2000
0.1500
0.1000
0.0500
0.0000
0 20 40 60 80
-0.0500
Time/(min)
Fig. 1 Weight of hexane as a function of time of the first step
in the rotary evaporator procedure (programmed vacuum
pressure=352 mbar)
time must be reproduced in every experiment to avoid
the positive or negative interferences that are incurred
around it; and two, and most important, is the long
time required for complete evaporation, which ele-
vates the energetic costs of the overall procedure.
In the set of experiments (2), other conditions were
investigated to achieve the quantitative evaporation
while reducing the total time in the rotary evaporator.
After each 10-min time step the distilled hexane was
removed from the receiving flask of the evaporator
system to alleviate vapour saturation. Decreasing the
programmed vacuum pressure from 352 mbar, recom-
mended for hexane, down to 200 mbar, was found to
achieve high evaporation yields (total “diesel recoveries”
of 107–116%). However, quantitative evaporation was
only accomplished after implementing an additional
vacuum system in the final desiccation step (100.3%
diesel recovery). The optimal conditions found for the
applied vacuum were 30 min at 22.5 in Hg (=0.75 atm),
but the procedure was not found to be too sensitive to the
vacuum time, since increasing it to 60 min yielded only a
1.1% diesel loss (i.e., 98.9% diesel recovery).
Thus, an approximate 30 min total vacuum time is
enough to achieve complete evaporation of hexane,
but the procedure does not require a very strict control
to adhere to this exact time. This makes it a very
practical procedural step, while at the same time
reducing rotary evaporator time costs to half of what
was necessary in the previously found conditions [set
of experiments (1)]. Also, it avoids the use of a drying
oven as a final step (EPA Method 1664 A 1999;
NMX-AA-134-SCFI-2006), which decreases energetic
Water Air Soil Pollut (2008) 194:151–161
costs, and it optimizes the desiccation step by, simulta-
neously: cooling, desiccating external adhered water,
and evaporating final hexane fractions. Finally, repro-
ducibility was considerably improved with this final
evaporation method [relative standard deviations (RSD)
of 0.2–0.3%], as compared to all previous experiments
(RSD of 2–7%).
100 120 140 3.2 Natural Organic Matter Co-extraction
and Cleanup
The fraction and nature of natural organic matter
(NOM) extracted from the standard soils by the
hexane treatment was investigated. NOM is responsi-
ble for most of the sorption phenomena that petroleum
hydrocarbons experience when equilibrating with soils
presumably via a homogeneous distribution through-
out the entire volume of NOM, in an analogous process
to solvent partitioning (Essington 2004). Due to its
low polarity, hexane is expected to extract a low
fraction of this NOM, which contains a variety of
polar functional groups (Essington 2004), however
even low fractions in absolute terms may represent
high relative gravimetric errors in the final determi-
nation of extracted TPH.
3.2.1 Quantification of NOM Extracted by Hexane
Quantification was performed on the clean soils
described in Section 2.2, with NOM contents ranging
from 0.5 to 4.6% organic carbon. The general
extraction procedure was followed (cf. Section 2) at
two different ultrasound power conditions (100 and
260 W), for a period of 25 min. The total hexane
volume after extraction and filtration was evaporated
according to the optimal conditions found in the
previous section. Additionally, two soils from the
Tabasco region in Southeast Mexico(courtesy of Dr.
Rutilio Ortiz, LAFQA, Geography Institute, UNAM),
with very high organic matter contents were included
in the experiments: Soils T1 (pH 4.8, 40.1% organic
carbon), and T2 (pH 3.6, 78.6% organic carbon). All
analyses were performed in triplicate.
Table 3 shows that for soils with OC contents
below 5%, extracted NOM was found below or barely
at the detection limit of the method (150 mg/kg) (thus
explaining the high relative errors found). From these
experiments, the potential co-extraction of NOM from
soils with hexane was not deemed interfering with the
Water Air Soil Pollut (2008) 194:151–161
Table 3 Average concentrations (mg/kg) of NOM extracted by n-hexane from non-contaminated soils at two ultrasound power
conditions, for 25 min
Soil
Ultrasound Power (W) F V
100 93
260 180
Relative SD (%) 30 and 32, respectively
a
These soil were obtained in a later phase of the experimental procedure, in which a higher ultrasound power was deemed necessary,
therefore their extraction was performed only at the higher power.
determination of contaminating TPH at a quantifica-
tion limit of 500 mg/kg.
However, in soils with very high NOM, extracted
fractions are considerable and depend on the total
NOM content. If no selective cleanup of these co-
extracted fractions is performed, they will necessarily
raise the effective TPH quantification limits on these
soils. For example, for soil T1 the DL would be
1.44 g/kg and the QL 4.80 g/kg, while for T2 these
would amount to 0.72% and 2.40% in mass,
respectively (see DL and QL definitions in Section 2).
3.2.2 Extract Cleanup Using Silica-gel
Silica-gel, according to EPA methods, is utilized to
eliminate interferences that are co-extracted with the
analytes. It is not clear what the actual cleanup
mechanism is since the hydroxyl groups at the silica
gel surface tend to be deprotonated and thus negatively
charged (Langmuir 1997) and would in principle
show repulsion for predominantly negatively charged
NOM functional groups (Essington 2004). Nonethe-
less, the use of silica-gel as a sorbent of interferent
compounds is widespread in standard methods that
quantify hydrocarbon contamination, and its efficacy
will be investigated here.
According to results shown in the previous
subsection, and given that NOM does not vary widely
in composition across soils in this range (Essington
2004), this cleanup procedure would only be neces-
sary for soils with total natural OC contents higher
than 5%. The following experiments were designed to
quantify the effectiveness and errors involved by
introducing such a cleanup step in the overall
gravimetric method.
Diesel Recovery After Eluting Through a Silica-Gel
Packed Column Accurately weighed 0.6 g of diesel
157
S R T1a T2a
108 30 75 – –
160 – – 480 2,400
34 and 33, respectively 33 23 – –
were mixed with 35 ml of hexane. The mixture was
eluted through a column packed with cotton, silica
gel, and celite, according to the description in the
general method, and subsequent hexane washings
through the column were applied: one of 5 ml, and
two of 10 ml (i.e., at the end, the extract contained a
total of 60 ml hexane). The hexane was evaporated
according to the optimal conditions established above.
A 0.3% diesel loss was registered. This corresponds
to a negligible concentration of 180 mg/kg loss from a
hypothetical 10 g soil (i.e., barely above the detection
limit of 150 mg/kg, and below the quantification limit
of 500 mg/kg—cf. Section 2.3), and was deemed
acceptable.
Efficiency of Silica Gel for Eliminating Co-Extracted
Soil NOM Accurately weighed 0.0025 g of NOM
extracted from the Vertisol (V) soil in the experiment
of the previous section were mixed with 0.0103 g of
diesel (accurately weighed) in 35 ml of hexane. These
proportions correspond to a hypothetical extraction of
10 g of soil containing 1 030 mg/kg TPH, from which
250 mg/kg NOM was co-extracted, which represents
an amount intermediate from soils with total OC
contents between 5% and 40%. The mixture was
transferred through a column packed according to the
general method, and washed with additional 25 ml of
hexane. The total hexane was evaporated according to
the optimal conditions found before.
A total mass recovery of 0.0120 g was registered
from the total 0.0128 g added, i.e. a 0.0018 g loss. If
we consider that this loss pertains exclusively to co-
extracted soil NOM retained, it would correspond to
(0.0008/0.0025×100=) 32% retention by the silica
gel. This amount represents a very low retention
efficiency, as was expected. To find out possible
reasons for this low experimental efficiency of silica-
gel as an adsorber of hexane-extracted soil NOM, a
158 Water Air Soil Pollut (2008) 194:151–161

more theoretical investigative approach was followed
by comparing the polar nature of this co-extracted
fraction with that of TPH standards used, through
gas chromatography with flame ionization detection
(GC-FID).
Brief Chromatographic Investigation of the Nature of
Hexane-Extracted Soil NOM Figure 2 shows the
corresponding gas chromatograms of soil extracts
and standards. The non-contaminated soil extracts
shows bands and peaks that appear in the 13 to
24 min time range. Figure 2a shows a chromatogram
example of such extracts for soil F. This same pattern
was confirmed on the extracts of other non-contaminated
soil samples listed in Table 2 (chromatograms not
shown). The bands and peaks observed overlap with
those of the diesel (Fig. 2b) and the diesel/lubricating
oil mixture standards (not shown), which appear in
the same region. The nature of the compounds
detected in this time range is of very low polarity or

Fig. 2 GC-FID chromatograms of hexane extracts from non-contaminated soils and of TPH standards [relative intensity (mV) vs.
time (min)], non-contaminated F soil (a), PEMEX diesel standard (b)
Water Air Soil Pollut (2008) 194:151–161 159

altogether non-polar, and the results suggest that at Table 4 Recovery of diesel from artificially contaminated
standard soils ‘V’ and ‘F’ by two different extraction procedures
least a fraction of the extracted soil NOM is
composed of compounds with similar chemical nature Soil Contamination % Recovery Relative
as the diesel compounds. This explains why these code level (mg/kg) average difference (%)
NOM compounds are co-extracted by hexane from
Soxhlet extraction
soils with TPH, why a major fraction of co-extracted
V1 1,018 103 3
NOM is not retained by silica, and confirms their V2 2,135 87 2
potential positive interference in the gravimetric F1 1,018 90 3
determination of petroleum-contaminated soils, if F2 2,135 89 2
high enough quantities are co-extracted (for example Ultrasound extraction bath at 100 Watts for 25 min SD (%)
from soils with very high NOM). F1 1,018 72.1 0.2
From these results it was decided that silica gel F2 2,135 68 1
F3 20,103 85.7 0.3
cleanup was not efficient and could be avoided in the
F4 100,112 90 3
overall method. However, this poses limitations of the
gravimetric method on the quantification limits for
soils with very high NOM contents. For example, for
soils with natural OC contents of 40%, in the absence not under investigation; these recovery figures will be
of silica gel cleanup, the DL would be approximately used solely as references for the ultrasound extraction
one order of magnitude higher than the DL for more recoveries.
average soils with OC contents below 5%. For soils TPH extractions following the general method
with near 80% OC, the DL would be approximately were performed using ultrasound baths at 100 W,
50 times higher. and at 260 W, using soil F. The results for the 100-W
bath are shown in Table 4 for the four levels of
3.3 TPH Extraction Efficiency Using Ultrasound contamination, and for 25 min extractions in triplicate.
Notice the low extraction efficiencies for F1 and F2, as
Soxhlet extractions of artificially-contaminated stan- compared to the Soxhlet extraction (Table 4). Figure 3
dard soils was performed to serve as reference for the shows the results for both ultrasound powers, for soils
ultrasound extraction investigations, by computing F1 and F2, and as a function of extraction time,
the maximum extractable fractions in these. The performed in triplicate. The results in this figure were
procedure was performed for samples V1, V2, F1, normalized to those obtained for the Soxhlet method
and F2. After extraction, the optimized hexane (Table 4), considering that the latter represent the
evaporation procedure described in Section 3.1 [set maximum extractable amount of diesel in the con-
of experiments (2)] was performed. ditions of the aged contaminated soils.
The results of these extractions (in duplicate) are Figure 3 shows a considerable direct effect of the
reported in Table 4 after correction from the blank, extraction time and ultrasound power on the total
but not corrected for possibly co-extracted NOM. TPH extraction. However, an increase in the hexane
Complete recovery of the diesel used as contaminant extractant volume from 35 to 45 ml did not yield
was obtained only for soil V1, the remaining soils improved extraction efficiencies. Extraction times
showed recoveries close to 90%. Since no measure- between 40 and 60 min were chosen as optimal;
ments, and thus no corrections, were made for co- however, under these conditions, it must be noted that
extracted NOM, the real diesel extraction efficiencies the recoveries obtained are somewhat lower by a
are most probably somewhat lower than the values range comprising 5–20% than those of the Soxhlet
reported in Table 4. extraction.
These apparently incomplete recoveries may be a In a final attempt to improve the ultrasound
result of diesel losses during preparation of the extraction efficiencies, the effect of sequential extrac-
contaminated standards, and/or presence of diesel tions was investigated, as suggested in standard
fractions that have become non-extractable with methods. The above extraction method was repeated
aging. However, the reasons for incomplete recovery three consecutive times on the same flask containing
do not concern us here, since the Soxhlet method is the initial 10 g of soil sample (V1 and V2 only). After
160
Soil F1 - 260 Watts
100 Soil F1 - 260 Watts, 45 mL
Soil F1 - 100 Watts
Soil F2 - 260 Watts
Soil F2 - 260 Watts, 45 mL
95 Soil F2 - 100 Watts
% Extraction efficiency
Soil V1 - 260 Watts
Soil V2 - 260 Watts
90
85
80
75
70
20 25 30 35 40
Ultrasound extraction time (min)
Fig. 3 Ultrasound extraction efficiencies of diesel-contaminated
soils F and V as a function of time and ultrasound power,
normalized to extraction data of an 8-h Soxhlet extraction
the first and second extractions, the major portion of
the hexane + hydrocarbon extract was carefully
decanted to the round-bottom flasks for evaporation,
through a cotton-packed glass column, but ensuring
that no soil particles were transferred to the columns.
Additional 25 ml of hexane were added to clean the
columns, making a total volume of 60 ml after each
consecutive extraction, which were collected sepa-
rately. After the third extraction, the whole soil was
decanted to the column and both the original flask and
the column were sequentially washed with 25
additional milliliters of hexane. The weights after
evaporation of the three flasks for each sample were
added together to compute total extraction efficiency.
No improvement was obtained upon three sequen-
tial extractions of the two soil samples, as compared
to one single extraction, which means that the first
extraction removed the whole ultrasound-extractable
fraction of diesel sorbed to the soils, and the
following two extractions did not contribute to any
additional extracted analyte. This was not immediately
obvious because of the nature of the experiment, where
remnants of the first extraction contributed to the
second and third extracted fractions. Thus, extraction
efficiencies for these two soils ranged from 80 to 96%
of the amounts extracted by the Soxhlet procedure
using the high-power ultrasound bath (Fig. 3). These
fractions may be regarded as representing those that
may become potentially available and eventually
hazardous to the biota and humans, in contrast to
the non ultrasound-extractable fractions.
Water Air Soil Pollut (2008) 194:151–161
4 Conclusions
The procedure of the developed gravimetric method
for TPH in soils in the present work may be
summarized as follows: (1) Accurate weighing of
approximately 10 g homogenized soil in round flasks,
and addition of 10 g of anhydrous Na2SO4; (2) TPH
extraction using 35 ml n-hexane in a 260-W ultra-
sound water bath for 60 min; (3) filtration of the
extracts through a column packed with 0.6 g treated
45 50 55 60 65
cotton or glass fibre, and down-washing with addi-
tional 25 ml hexane to complete 60 ml in the final
liquid extract for analysis; (5) evaporation of the
hexane in three 10-min steps in a rotary evaporator at
40°C, followed by quantitative evaporation of the
remnant hexane in a desiccator under 22.5 in Hg
(0.75 atm) vacuum for 30 min; and (6) weighing of
the residue in an analytical balance.
The novel aspects of the method presented relate to
cost- and time-saving steps. These are reflected in the
extraction procedure by using a relatively low volume
of hexane extractant and an ultrasound bath for 1 h,
instead of the 4 to 8-h Soxhlet extraction recommended
in standard methods; also, the overall use of a heating
oven was minimized, since evaporation of the solvent
(and of possible water adsorbed to the flask wall) is
completed using vacuum (inside a desiccator). Method
expediency was achieved by eliminating a silica-gel
cleanup procedure, which was justified by independent
experiments that showed that silica-gel does not
selectively retain co-extracted natural organic matter.
Insufficient solvent evaporation was identified as one
of the most important positive interferences in the
gravimetric detection in general, and it is highly
recommended to carefully calibrate this step in any
method modification, especially as a function of type
and total extractant volume. Quantitative hexane
evaporation via the desiccation under vacuum step
investigated was not found to be highly sensitive to
time and thus allowed for method convenience.
However, the method was found to be positively
sensitive to the extraction time and power of the
ultrasound bath used, and these parameters should also
be carefully calibrated in any method modification.
Acknowledgements The authors are grateful to: Dr. Arturo
Aguirre Gómez from the Department of Chemistry at FES-
Cuautitlán, UNAM for supplying the soils of study, for the pH
Water Air Soil Pollut (2008) 194:151–161
and electric conductivity data for these, and for his assistance in
the organic carbon determinations; Dr. Rutilio Ortiz, Geogra-
phy Institute, UNAM, for supplying the high organic matter
content soils, and for the pH and organic carbon data for these;
Rosaura Paez, Geography Institute, UNAM, Morelia Unit, for
help in acquiring the gas chromatograms; Ricardo Alfaro,
Chemistry Institute, UNAM, for his assistance in the initial
methods development; and two anonymous reviewers who
helped improve considerably the legibility of the manuscript.
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