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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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Margarita Eugenia Gutirrez ruiz
Universidad Nacional Autnoma de Mxico
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Water Air Soil Pollut (2008) 194:151161

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 identified as the most prevalent and largest positive
interference in the gravimetric detection. To overcome
this, while minimizing the need for heating, a combination 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 4060 min treatment in an

ultrasound bath of 260 W was found suitable to extract
8095% 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.

M. Villalobos (*) : M. E. Gutierrez-Ruiz

Grupo de Bio-Geoqumica Ambiental,
Laboratorio de FisicoQumica Ambiental (LAFQA)
Instituto de Geografa,
Universidad Nacional Autnoma de Mxico (UNAM),
Coyoacn 04510 D.F., Mxico
e-mail: marvilla@igg.unam.mx

1 Introduction

M. E. Gutierrez-Ruiz
e-mail: ginny@servidor.unam.mx
A. P. Avila-Forcada
Direccin General del Centro Nacional de Investigacin y
Capacitacin Ambiental (CENICA),
Instituto Nacional de Ecologa, San Rafael Atlixco 186,
Col. Vicentina,
Iztapalapa. D.F. 09340, Mxico, Mxico
e-mail: apavila@ine.gob.mx

Keywords Contaminated soils . Gravimetric method .

Silica gel . Solvent evaporation . Total petroleum
hydrocarbons . Ultrasound extraction

Soil pollution by petroleum hydrocarbons usually

originates from spills or leaks of storage tanks during
fuel supply and discharge operations. Most petroleum
components are hazardous to human health and to the
soil biota (MDEP 1994); therefore, measurement of their
total concentration (total petroleum hydrocarbons
TPH) is the initial and most general indicator of hazard
by hydrocarbons for environmental diagnostic purposes.
The US Environmental Protection Agency (USEPA)
has published a series of methods for quantifying TPH
in solid matrices (including soils; Table 1). The four
categories of analytical methods described in Table 1
are based on the final detection method, i.e. gravimetry,

152

Water Air Soil Pollut (2008) 194:151161

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

EPA method
Extracting solvent and
no. (year of most range of hydrocarbon
recent revision) species extracted

Approximate Aprox. Cost/ Advantages

detection limit, samplea
mg/kg (soil)

Gravimetric

9071 B (1998)
1664 A, for
water samples
(1999)

n-Hexane. Most
compounds except
volatile fractions

50

50

Infrared (IR)

8440 (1996)

Supercritical CO2.
Most compounds
except volatile and
very heavy fractions

10

5080

Gas chromatography 8015 C (2000)

(GC)

Immunoassay

4030 (1996)

10
Methanol, n-hexane,
2-propanol,
ciclohexane,
acetonitrile, acetone,
methylene chloride, or
mixtures of these.
Normally between C6
and C25 or C36 (may
be modified for a higher
range)
The extractant depends 10500
on the commercial kit.
Aromatic hydrocarbons
(BTEX, PAHs)

100150

13b
64c

Disadvantages

Simple, quick,
Lack of specificity, low
and inexpensive sensibility, high loss of
volatiles, prone to
interferences; only
useful for total
quantification.
Simple, quick,
Lack of specificity, low
and inexpensive sensitivity, high loss of
volatiles, poor
extraction of high
molecular weight
hydrocarbons, prone to
interferences; only
useful for total
quantification
Detects a broad Normally it cannot detect
compounds below C6;
range of
does not detect polar
hydrocarbons
hydrocarbons (alcohols,
compounds;
ethers, etc.); chlorinated
simple and
solvents may be
sensitive;
quantified as TPH
chemical
speciation is
possible.
Simple, quick,
inexpensive,
and may be
used in the field

Low sensitivity, prone to

interferences, primarily
measures aromatics; low
accuracy and precision;
must be used as a
screening method; only
useful for total
quantification

US dollars, MDEP (1994)

CMECC (1996)

VRAI (2007)

infrared spectroscopy, gas chromatography, and immunoassays. Currently, Mexican legislation has implemented many of these as standard methods for
diagnosis of contaminated soil sites, especially methods 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 measurement. 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 procedures 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) 194:151161

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 investigated 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, ASTM2 mm,
followed by Sieve 200, ASTM75 m), and dried to
105C 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 (105C) to constant weight. Ten grams of
anhydrous Na2SO4 were added to create a freeflowing 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 evaporation of the remnant hexane under variable conditions (cf. Section 3). The residue was weighed in an
analytical balance and designated as TPH.
The equipment and material used were: a semimicroanalytical 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=40C, Cooling water
temperature=14C, Rotation speed=70 rpm, automatic
mode. Subsequently the flasks were dried with absorbent 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 length10 mm internal diameter.
The reagents employed were Baker analytical
reagents (n-hexane, acetone, silica gel 40140, and

154

Water Air Soil Pollut (2008) 194:151161

anhydrous sodium sulfate). Celite (Qumica 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).

volume in the rotary evaporator and filtered through a

cotton-packed column, washing with additional hexane to make a total volume of 60 ml. The procedure
described in the general method was followed from this
point onwards.

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).

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 400C).
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 homogenized 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 concentrations shown below, which represent a wide range

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 150C 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 150C 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 performed for 8 h to ensure maximum recovery, after
which the extract was concentrated to 40-ml total

2.2 Preparation of Standard Soils Contaminated

with TPH

Table 2 Main characteristics of non-contaminated soils used in the preparation of TPH-contaminated soil standards
Code

Soil descriptor

pH

Electric conductivity (mS/cm)

% Organic carbon (OC)a

F
S
V
R

Fertilized
Sodic saline
Vertisol
Red

7.7
9.5
7.1
7.2

0.730
1.322
1.500
1.224

1.72
0.46
4.56
0.57

Soils provided by courtesy of Dr. Arturo Aguirre Gmez, Department of Chemistry, FES Cuautitln, UNAM.
a

Analysed in triplicate according to Aguirre et al. (2002)

Water Air Soil Pollut (2008) 194:151161

155

of TPH soil contamination levels (from 0.1% to 10%

in weight):

(rate, 6C/min) to 300C (rate, 4C/min); total time,

49 min.

Soil number code

Applied diesel concentration
(mg/kg)

3 Results and Discussion

1
2
3
4
5
1,018 2,135 20,103 100,110 0

In the following section the details of the procedures carried out and the parameters studied to
calibrate the method are described in the order in
which they were performed, providing proper sensitivity 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 nonhexane-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+oilcontaminated 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 m) column;
nitrogen carrier gas (1 ml/min); flame ionization
detector temperature, 320C; furnace program, 70C
(isotherm, 0.5 min) to 125C (rate, 8C/min) to 245C

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 its 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 105C flasks that were not previously ovendried (error<0.0007 g), or from previously oven-dried
flasks at 105C 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 investigated: (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

Water Air Soil Pollut (2008) 194:151161

Final hexane weight/(g)

0.2500

costs, and it optimizes the desiccation step by, simultaneously: cooling, desiccating external adhered water,
and evaporating final hexane fractions. Finally, reproducibility was considerably improved with this final
evaporation method [relative standard deviations (RSD)
of 0.20.3%], as compared to all previous experiments
(RSD of 27%).

0.2000
0.1500
0.1000
0.0500
0.0000
0

20

40

60

80

100

120

140

-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 elevates 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, recommended for hexane, down to 200 mbar, was found to
achieve high evaporation yields (total diesel recoveries
of 107116%). 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

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 responsible for most of the sorption phenomena that petroleum
hydrocarbons experience when equilibrating with soils
presumably via a homogeneous distribution throughout 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 determination 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) 194:151161

157

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)
100
260
Relative SD (%)

F
93
180
30 and 32, respectively

V
108
160
34 and 33, respectively

S
30

33

R
75

23

T1a

480

T2a

2,400

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 quantification 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 coextracted 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). Nonetheless, 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 necessary 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

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/kgcf. 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 coextracted soil NOM retained, it would correspond to
(0.0008/0.0025100=) 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 silicagel as an adsorber of hexane-extracted soil NOM, a

158

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

Water Air Soil Pollut (2008) 194:151161

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) 194:151161

altogether non-polar, and the results suggest that at

least a fraction of the extracted soil NOM is
composed of compounds with similar chemical nature
as the diesel compounds. This explains why these
NOM compounds are co-extracted by hexane from
soils with TPH, why a major fraction of co-extracted
NOM is not retained by silica, and confirms their
potential positive interference in the gravimetric
determination of petroleum-contaminated soils, if
high enough quantities are co-extracted (for example
from soils with very high NOM).
From these results it was decided that silica gel
cleanup was not efficient and could be avoided in the
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
of silica gel cleanup, the DL would be approximately
one order of magnitude higher than the DL for more
average soils with OC contents below 5%. For soils
with near 80% OC, the DL would be approximately
50 times higher.
3.3 TPH Extraction Efficiency Using Ultrasound
Soxhlet extractions of artificially-contaminated standard soils was performed to serve as reference for the
ultrasound extraction investigations, by computing
the maximum extractable fractions in these. The
procedure was performed for samples V1, V2, F1,
and F2. After extraction, the optimized hexane
evaporation procedure described in Section 3.1 [set
of experiments (2)] was performed.
The results of these extractions (in duplicate) are
reported in Table 4 after correction from the blank,
but not corrected for possibly co-extracted NOM.
Complete recovery of the diesel used as contaminant
was obtained only for soil V1, the remaining soils
showed recoveries close to 90%. Since no measurements, and thus no corrections, were made for coextracted NOM, the real diesel extraction efficiencies
are most probably somewhat lower than the values
reported in Table 4.
These apparently incomplete recoveries may be a
result of diesel losses during preparation of the
contaminated standards, and/or presence of diesel
fractions that have become non-extractable with
aging. However, the reasons for incomplete recovery
do not concern us here, since the Soxhlet method is

159
Table 4 Recovery of diesel from artificially contaminated
standard soils V and F by two different extraction procedures
Soil
code

Contamination
level (mg/kg)

% Recovery
average

Relative
difference (%)

Soxhlet extraction
V1
1,018
103
3
V2
2,135
87
2
F1
1,018
90
3
F2
2,135
89
2
Ultrasound extraction bath at 100 Watts for 25 min SD (%)
F1
1,018
72.1
0.2
F2
2,135
68
1
F3
20,103
85.7
0.3
F4
100,112
90
3

not under investigation; these recovery figures will be

used solely as references for the ultrasound extraction
recoveries.
TPH extractions following the general method
were performed using ultrasound baths at 100 W,
and at 260 W, using soil F. The results for the 100-W
bath are shown in Table 4 for the four levels of
contamination, and for 25 min extractions in triplicate.
Notice the low extraction efficiencies for F1 and F2, as
compared to the Soxhlet extraction (Table 4). Figure 3
shows the results for both ultrasound powers, for soils
F1 and F2, and as a function of extraction time,
performed in triplicate. The results in this figure were
normalized to those obtained for the Soxhlet method
(Table 4), considering that the latter represent the
maximum extractable amount of diesel in the conditions of the aged contaminated soils.
Figure 3 shows a considerable direct effect of the
extraction time and ultrasound power on the total
TPH extraction. However, an increase in the hexane
extractant volume from 35 to 45 ml did not yield
improved extraction efficiencies. Extraction times
between 40 and 60 min were chosen as optimal;
however, under these conditions, it must be noted that
the recoveries obtained are somewhat lower by a
range comprising 520% than those of the Soxhlet
extraction.
In a final attempt to improve the ultrasound
extraction efficiencies, the effect of sequential extractions was investigated, as suggested in standard
methods. The above extraction method was repeated
three consecutive times on the same flask containing
the initial 10 g of soil sample (V1 and V2 only). After

160

% Extraction efficiency

100
95

Water Air Soil Pollut (2008) 194:151161

Soil F1 - 260 Watts
Soil F1 - 260 Watts, 45 mL
Soil F1 - 100 Watts
Soil F2 - 260 Watts
Soil F2 - 260 Watts, 45 mL
Soil F2 - 100 Watts
Soil V1 - 260 Watts
Soil V2 - 260 Watts

4 Conclusions

90
85
80
75
70
20

25

30
35
40
45
50
55
Ultrasound extraction time (min)

60

65

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 separately. 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 sequential 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.

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 ultrasound water bath for 60 min; (3) filtration of the
extracts through a column packed with 0.6 g treated
cotton or glass fibre, and down-washing with additional 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
40C, 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 Gmez from the Department of Chemistry at FESCuautitln, UNAM for supplying the soils of study, for the pH

Water Air Soil Pollut (2008) 194:151161

and electric conductivity data for these, and for his assistance in
the organic carbon determinations; Dr. Rutilio Ortiz, Geography 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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