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Analyst, August 1998, Vol. 123 (1679–1684)
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1679
Comparison of microwave-assisted extraction and
Soxhlet extraction for phenols in soil samples using
experimental designs
A. Egizabal, O. Zuloaga*, N. Etxebarria, L. A. Fernández and J. M. Madariaga
Kimika Analitikoaren Saila, Euskal Herriko Unibertsitatea, 644 P.K., E-48080 Bilbao, Spain
A study and comparison of microwave-assisted extraction
(MAE) and Soxhlet extraction for five phenols (phenol,
2-chlorophenol, 2-methylphenol, 2-nitrophenol and
2,4-dichlorophenol) in soils was carried out in order to
establish a comparative analysis between the two
techniques under their optimum conditions. The
optimisation of both the MAE and Soxhlet extraction
methods was achieved using experimental designs. The
parameters studied were pressure inside the extraction
vessel, extraction time, percentage of microwave power,
solvent volume and percentage (v/v) of acetone in an
acetone–hexane mixture for MAE and extraction time and
percentage (v/v) of acetone in an acetone–hexane mixture
for Soxhlet extraction. The simplex method was also used
for obtaining the optimum conditions in some cases. The
extracts were analysed by GC–MS or GC–FID.
Keywords: Microwave-assisted extraction; Soxhlet extraction;
gas chromatography–mass spectrometry; gas
chromatography-flame ionization detection; phenols;
experimental designs; simplex method; soil samples
Phenol and its derivatives are widely used in the chemical
industry for the manufacture of polymers, textiles, drugs, resins,
dyes, detergents, explosives, stabilisers and antioxidants.1
Further, phenolic compounds have substantial applications in
agriculture as herbicides, insecticides and fungicides, thus
becoming potential pollutants of soils and of surface and
underground waters owing to their highly hydrophilic nature.2
However, phenolic compounds are not only generated by
human activity, they are also formed naturally, e.g., during the
decomposition of leaves or wood.2
Phenols have a highly toxic character and it is well known
that these substances exhibit properties that are hazardous to
human health.3 Owing to their toxicity and presence in the
environment, the US Environmental Protection Agency (EPA)
includes 11 phenols among the main environmental pollut-
ants.4
The extraction of organic pollutants from soil samples
requires the use of organic solvents, which compete in the
release of the analytes retained owing to the high activity of the
matrix. Traditional methods employ large volumes of solvents
under aggressive shaking and/or temperature conditions. The
most frequently used method for the extraction of organic
pollutants from soils is extraction at the boiling-point of the
solvent (Soxhlet extraction) or the use of an ultrasonic bath.
Soxhlet extraction is the most conventional of all methods and
consists of a simple distillation process repeated a number of
times. This method is particularly suitable for organic pollutants
strongly adsorbed on soil matrices but requires long extraction
times and the use of large volumes of frequently toxic solvents.
Moreover, it can degrade the analytes.5,6
In the last few years, the number of procedures using
extraction of organic compounds from environmental matrices
by microwave energy has increased.7–11 Microwave energy is a
non-ionizing radiation that causes molecular motion by migra-
tion of ions and rotation of dipoles, without changing the
molecular structures if the temperature is not too high. Non-
polar solvents, such as hexane and toluene, are not affected by
microwave energy and, therefore, it is necessry to add polar
additives.7 Microwave extraction greatly reduces solvent con-
sumption and extraction times. Many of these studies7–9 made
use of the experimental design approach for the optimisation of
microwave-assisted extraction (MAE) procedures. The varia-
bles considered differ from one publication to another, but the
most commonly used combinations are temperature, extraction
time and solvent volume,7 volume of solvent, percentage of
acetone and extraction temperature8 of different types of
extracting solvents, extraction temperature, extraction time and
solvent volume.9
In this work, MAE was used for the extraction of five
phenolic compounds, phenol, 2-chlorophenol, 2-methylphenol,
2-nitrophenol and 2,4-dichlorophenol and the results were
compared with those obtained by Soxhlet extraction. The
parameters studied for MAE were pressure inside the extraction
vessel, extraction time, percentage of microwave power,
percentage (v/v) of acetone in the acetone–hexane mixture and
volume of solvent. For Soxhlet extraction, the percentage (v/v)
of acetone in the acetone–hexane mixture and extraction time
were studied. Factorial and central composite designs were
mainly used for approaching the response surface but the
simplex method12 implemented in the MultiSimplex program13
was also used in some cases for achieving the optimum
conditions.
Experimental
Microwave-assisted extraction
MAE experiments were performed with an MDS-2000 closed
microwave solvent extraction system (CEM, Matthews, NC,
USA) equipped with a 12-sample tray and pressure feedback/
control. A 0.5 g aliquot of spiked soil was accurately weighed,
mixed with 2 g of anhydrous sodium sulfate and transferred
quantitatively to the Teflon-lined extraction vessel. The ace-
tone–hexane mixture was then added to each sample and the
extraction vessel was closed, after ensuring that a new rupture
membrane was used for each experiment. In this work, the
volume and percentage (v/v) of acetone in the acetone–hexane
mixture and the power, pressure and time in which extractions
were performed were indicated by the experimental design. The
extraction conditions were programmed in two stages. In the
first stage, the system was allowed to reach the required
pressure using full power; in the second stage, the pressure
previously reached was kept constant and the power and the
time were established according to the experimental design. In
this way, the samples stayed at the previously set pressure
during the whole extraction process. When the irradiation
period was complete, samples were removed from the micro-
wave cavity and allowed to cool to room temperature before
opening. The supernatant was filtered through glass-wool pre-
washed with hexane–acetone and then combined with 7–10 ml
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1680 Analyst, August 1998, Vol. 123

acetone–hexane rinses of the extracted sample. The extract was
concentrated to approximately 1 ml using nitrogen evaporation
with a Turbovap LV Evaporator (Zymark, Hopkinson, MA,
USA). An equivalent amount of a stock standard solution of
phenols followed both the filtration and concentration steps in
order to estimate possible losses occurring in these steps. The
concentrated extracts, together with 200 ml of a stock standard
solution of 4-methoxyphenol (internal standard), were trans-
ferred into 5 ml calibrated flasks and dissolved in hexane. The
extracts were transferred into injection vials for GC–FID or
GC–MS analysis.
Soxhlet extraction
Soxhlet extractions were performed using 1 g of soil analytic-
ally weighed and mixed with 10 g of anhydrous sodium sulfate.
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from LabScan (Dublin, Ireland) and the purities were stated to
be better than 95%. Anhydrous sodium sulfate (purissimum)
was purchased from Panreac (Barcelona, Spain). All solutions
were stored at 5 °C in the dark. All volumetric glassware was
grade A and was calibrated at laboratory temperature.
Optimisation experiments were performed using a clay soil
from an industrial downfall (Metalquı́mica del Nervión, Bilbao,
Spain). The sample was ground, sieved and dried following the
ISO 11464 Norm14 for homogeneity. A 40 g portion of soil,
which was checked to be free of phenols, was analytically
weighed and 35 ml of 40 mg ml21 stock solution were added.
Subsequently, acetone was added to cover the soil and the
mixture was stirred for at least 24 h. The soil was then dried in
an oven at 40 °C. Finally, the soil was homogenised with a
mortar, bottled and stored in a refrigerator. The concentrations
in the soil, on the basis of added amounts, were 34.4, 40.6, 36.3,

Samples were extracted under reflux with 200 ml of acetone– 35.2, and 35.6 mg g21 for phenol, 2-chlorophenol, 2-methylphe-
hexane in the percentages and the times indicated by the nol, 2-nitrophenol and 2,4-dichlorophenol, respectively.
experimental design. After cooling to room temperature, the
extracts were concentrated to approximately 5 ml using a rotary Analysis of extracts
evaporator at 40 °C and reduced pressure. To estimate possible
losses occurring during the concentration stage, an amount of a Some extracts were analysed on a Hewlett-Packard (Avondale,
stock standard solution of phenols was treated in the same PA, USA) Model 5890 Series II gas chromatograph interfaced
manner. The concentrated extracts, together with 200 ml of a
stock standard solution of 4-methoxyphenol (internal standard),
were transferred into 10 ml calibrated flasks and dissolved in
hexane. The extracts were transferred into injection vials for
GC–FID or GC–MS analysis.

Reagents and chemicals

Phenol standards were supplied as follows: phenol and
4-methoxyphenol by Merck (Darmstadt, Germany), 2-chloro-
phenol and 2,4-dichlorophenol by Aldrich (Gillingham, Dorset,
UK) and 2-nitrophenol and 2-methylphenol by Fluka (Buchs,
Switzerland). All compounds were analytical-reagent grade and
the purities were stated to be higher than 99%. A stock standard
solution (40 mg ml21) of phenol, 2-chlorophenol, 2-methylphe-
nol, 2-nitrophenol and 2,4-dichlorophenol was prepared by
weighing an appropriate amount of the standard and dissolution
in 50 ml of hexane. This standard solution was used only for the
sample spiking. A stock standard solution (1000 mg ml21) of
4-methoxyphenol (internal standard) was prepared by weighing
an appropriate amount and dissolving it in 20 ml of hexane.
From the stock standard solution of the internal standard a more
dilute working standard solution (40 mg ml21) was prepared
daily and then used in all calibration standards and sample
extracts. For calibration, the mixture number 5 of 18 semi-
volatile phenols (2000 mg ml21) supplied by Supelco (Belle- Fig. 1 Sheward plots for phenol by (a) GC–MS and (b) GC–FID. Drf
fonte, PA, USA) was used. Hexane and acetone were purchased = average relative response factor.

Table 1 Design matrix and corrected recovery values in the fractionated factorial design for MAE experiments

Design matrix Recovery (%)

Run Pressure/ Time/ Power Acetone Volume/ 2-Chloro- 2-Methyl- 2-Nitro 2,4-Dichloro-
No. psi min (%) (%) ml Phenol phenol phenol phenol phenol
1 20 20 30 30 30 65 58 50 60 56
2 30 20 30 70 15 99 84 73 102 89
3 20 40 30 70 15 88 77 68 93 71
4 30 40 60 30 30 84 80 61 103 81
5 20 20 60 70 30 80 69 59 84 68
6 30 20 60 30 15 26 26 22 30 25
7 20 40 60 30 15 90 85 67 94 84
8 30 40 60 70 30 151 140 128 133 128
9 25 30 45 50 20 109 96 85 86 93
10 25 30 45 50 20 96 79 71 72 79
11 25 30 45 50 20 77 66 58 58 65
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Analyst, August 1998, Vol. 123 1681

to a Hewlett-Packard 5989 MS Engine mass spectrometer MS/
DOS ChemStation and equipped with a Hewlett-Packard Model
6890 Series autosampler. The samples were analysed on a 30 m
3 0.32 mm id 3 0.25 mm film thickness HP-5 fused-silica
capillary open-tubular column. The column temperature was
held at 40 °C for 2 min and then increased at 30 °C min21 to
80 °C, where it was held for 1 min and subsequently
programmed at 10 °C min21 to 200 °C, where it was held for 2
min. The carrier gas was helium (N-50) at a flow rate of 3.45
ml min21 and a pressure at the column head of 10 psi. The
injection volume was 1–3 ml and the injection temperature was
250 °C. The injector was set in the splitless mode with split vent
closed for 1 min after injection. For the mass spectrometer, the
electron energy was set at 70 eV and spectral data were acquired
at a rate of 0.8 scan s21 (scanning range 50–550 u). Quantitative
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data were acquired by operating the MS detector in the selected
ion monitoring mode (SIM) and the m/z values monitored were
phenol 94, 66, 2-chlorophenol 128, 64, 2-methylphenol 108,
107, 2-nitrophenol 139, 65, 4-methoxyphenol 124, 109 and
2,4-dichlorophenol 164, 162. The instrument was tuned daily
with perfluorotributylamine (PFTBA) using the EI Sensitivity
Tune (EISENS) internal program since this program allows
better sensitivities than those obtained by the usual Automatic
Tune (ATUNE). In addition, to increase sensitivity, an optimum
400 V overpotential was applied to the electronic multiplier. A
five-point internal standard calibration using standards at 0.32,
1.65 and 8 mg ml21 and injecting different volumes (1–3 ml) of
the standards was performed daily to establish the GC–MS
calibration curve, which ranged from 0.6 to 9.2 ng. 4-Methoxy-
phenol (internal standard) was added to every calibration

Table 2 Design matrix and corrected recovery values in the central composite design for MAE experiments

Design matrix Recovery (%)

Run Power Time/ Pressure/ 2-Chloro- 2-Methyl- 2-Nitro- 2,4-Dichloro-

No. (%) min psi Phenol phenol phenol phenol phenol
1 44 16.5 16 89 55 51 37 51
2 70 16.5 16 71 44 37 54 47
3 44 33.5 16 106 63 59 44 58
4 70 33.5 16 121 76 69 71 80
5 44 16.5 26 122 82 67 105 95
6 70 16.5 26 108 70 52 86 82
7 44 33.5 26 125 81 68 110 96
8 70 33.5 26 78 44 42 32 45
9 80 25 21 96 64 42 87 74
10 35 25 21 105 71 56 91 82
11 57 40 21 106 66 57 58 68
12 57 10 21 99 63 53 87 77
13 57 25 30 59 37 31 45 43
14 57 25 12 93 60 49 64 64
15 57 25 21 87 51 38 46 54
16 57 25 21 87 54 45 47 58
17 57 25 21 105 52 55 95 55

Table 3 Values of the parameters obtained for central composite design for MAE experiments

Compound b0 b1 b2 b12 b13 b23 b11 b22 b123 r2

Phenol 113 23.32 26.01 0.22 0.14 0.31 —* — 20.010 0.753
2-Chlorophenol 201 25.80 27.84 0.17 0.09 0.25 0.03 0.05 20.008 0.858
2-Methylphenol 112 22.13 26.45 0.13 0.07 0.18 — 0.04 20.006 0.742
2-Nitrophenol 308 29.95 216.22 0.27 0.12 0.67 0.07 0.09 20.015 0.805
2,4-Dichlorophenol 225 26.96 211.28 0.23 0.12 0.42 0.04 0.06 20.010 0.857
* Dashes indicate that the parameter was eliminated from the model.

(a) 50 (b) (c)

80
45
recovery (%)

recovery (%)

recovery (%)

45
70
40
40
60
35 35
50
30
30
40 40
10
50
pow

po 50 15
we 60 35 40 60 20 30
30 tim 25
er (

r ( 70 25
% 20 25 in 70 25
30 e/ 30
m 20 i
) 80 10 15 time/m e/ps
%)

20 in 35
80 re/psi 15 ressur
15 pressu 40 p
Fig. 2 Response surface for the central levels of (a) pressure (b) time and (c) power for the central composite design.
 View Article Online

1682 Analyst, August 1998, Vol. 123

Table 4 Design matrix and corrected recovery values in the complete factorial design for Soxhlet experiments

Design matrix Recovery (%)

Time/ Acetone 2-Chloro- 2-Methyl- 2-Nitro- 2,4-Dichloro-

Run No. min (%) Phenol phenol phenol phenol phenol
1 8 30 51 68 14 57 98
1 8 30 72 68 14 61 85
1 8 30 65 68 15 40 101
2 16 30 61 77 11 42 90
3 8 70 107 69 15 52 94
4 16 70 95 84 14 60 80

standard and sample extract that was analysed by GC–MS. Each The data analysis of the results given in Table 1 was
calibration standard and sample extract was injected in performed using the non-linear regression analysis program
triplicate. Chromatographic peak areas were fitted by linear NLREG.16 Response surfaces (Y) were taken as a function of
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regression and the correlation coefficients ranged from 0.998 to the considered variables (xi) using polynomials of different
0.999. For quantification, the average relative response factors degree depending on the experimental design followed. The
obtained from the multilevel calibration was used. In addition, general polynomial function is
the relative response factors obtained daily were checked
following Sheward representations.15 As can be deduced from
Fig. 1(a) for phenol, all data are under statistical control. All the
other phenols presented a similar pattern.
Y = b0 +
 i
b i xi +
Â
ij
b ij xi x j (3)

Other extracts were analysed on a Hewlett-Packard Model where Y is the recovery, xi are the variables considered for the
5890 Series II gas chromatograph equipped with a flame optimisation of the extraction and bi and bij are the parameters
ionisation detector. The chromatographic conditions were as to be calculated. The estimation of the parameters (bi and bij)
given above. The detector was kept at 300 °C. A five-point was achieved by the minimisation of the square sum of errors
internal standard calibration using standards of 1, 2, 4, 6 and 8 (U) as given by the equation
mg ml21 and always injecting the same volume (1 ml) of the
standards was performed daily to esablish the GC–FID n
calibration curve. 4-Methoxyphenol (internal standard) was
added to every calibration standard and sample extract that was
analysed by GC–FID. Each calibration standard and sample
U=
 i
(Yexp - Ycalc ) 2 (4)

extract was injected in triplicate. Chromatographic peak areas
were fitted by linear regression and the correlation coefficients
ranged from 0.983 to 0.999. For quantification, the average
relative response factors obtained from the multilevel calibra-
tion were used and their statistical control was checked through
the Sheward plot [Fig. 1(b)].
Results and discussion
Fractionated factorial design for MAE
The variables considered in the fractionated factorial design
were pressure of extraction, percentage of microwave power,
extraction time, volume of solvent and percentage (v/v) of
acetone in the acetone–hexane mixture. The aim of this design
was to evaluate which of the variables were factors, that is,
which of the variables had an influence on the extraction
process and which did not. Owing to the high number of
experiments for a two-level factorial design (25), two levels of
fractionality were introduced and the percentage of acetone and
the volume of the solvent were defined as a combination of the
other three variables (pressure, power and time) as shown in the
equations.
percentage of acetone M pressure 3 time 3 power
volume M pressure 3 time
The percentage of acetone and solvent volume were chosen
for the fractionality since they were a priori the variables which
least interacted with the other three (pressure, time and power).
A two-level fractionated factorial design involving eight runs
plus three central points was chosen. The design matrix and the
results for the 11 runs (which were measured by GC–MS and
whose percentage recoveries were corrected with the nitrogen
blowdown evaporation recovery) are given in Table 1.
The analysis of the output was based on the evaluation of the
prob(t) parameter associated with each bi parameter, since
prob(t) indicates the probability of bi being zero. Those
parameters whose probability of being zero was greater than
10%, i.e., prob(t) > 0.1, were systematically eliminated.
The most general function allowed for the fractionated
experimental design is
Y = b0 + b1x1 + b2x2 + b3x3 + b4x4 + b5x5 + b13x1x3 +
b23x2x3
(5)
where x1 is the pressure in the extraction vessel, x2 is the
extraction time, x3 is the percentage of microwave power, x4 is
the percentage (v/v) of acetone and x5 is the solvent volume. b12
and b24 were not considered because they were equivalent to b5
and b13, respectively. The reason for this equivalence lies in the
way variables x4 and x5 were defined (x5 M x1x2; x2x4 M
x1x3).
From the values obtained for the parameters for the
fractionated factorial design it could be concluded that all
variables [pressure, time, power, percentage (v/v) of acetone
and volume] have an influence in the extraction process. For all
phenols the parameters were of the same sign and magnitude
with the exception of 2-methylphenol, for which pressure
(1) seemed to have no effect (parameter b1 = b13 = 0).
(2)
Central composite design for MAE
From the fractionated factorial design, none of the variables
could be excluded because all of them [pressure, time, power,
percentage (v/v) of acetone and volume] seemed to have an
influence on the extraction process. Because of this, the
percentage of acetone in the acetone–hexane mixture and the
volume were fixed to at 50 : 50 (v/v) and 15 ml, respectively,
and a central composite design was built using the other three
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Analyst, August 1998, Vol. 123 1683

variables: pressure inside the extraction vessel, extraction time
and percentage of microwave power. A two-level factorial
design plus star orthogonal composite design involving 14 runs
plus three central points was chosen. Table 2 gives the design
matrix for this experiment and the net recoveries obtained in
each run.
The data in Table 2 were analysed by NLREG following the
methodology mentioned above. The most general function for
central composite design is
Y = b0 + b1x1 + b2x2 + b3x3 + b12x1x2 + b13x1x3 b23x2x3 +
b11x21 + b22x22 + b33x23 + b123x1x2x3 (6)
Table 5 Recoveries for 15 samples of 2-chlorophenol extracted for 3 days
by MAE
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where x1 is the percentage of microwave power, x2 is the
extraction time and x3 is the pressure inside the extraction
vessel. The results obtained by NLREG are given in Table 3.
From the output data the extraction recoveries were not directly
affected by pressure (parameters b3 = b33 = 0). Nevertheless,
this does not mean that pressure presented no effect during the
extraction since interaction parameters b13 (power–pressure).
b13 (time–pressure) and b123 (power–time–pressure) were
different from zero. As for the fractionated factorial design, all
parameters showed the same sign and magnitude for the five
phenols.
Three-dimensional representations keeping one of the varia-
bles at the central point value are presented in Fig. 2 for
2-chlorophenol. At constant pressure (P = 21 psi) a maximum
around 35–40% for power and 10–15 min for the extraction
time could be observed [Fig. 2(a)]. On the other hand, at time

= 25 min [Fig. 2(b)], the maximum was located at power = 40–

Day No. 50% and pressure = 15–20 psi. Finally, for power = 57%
Sample [Fig. 2(c)], a maximum appeared for pressure = 25–30 psi and
No. 1 2 3 time = 15–25 min. It could be concluded that the optimum
1 67 69 82 conditions were a low percentage of microwave power
2 78 66 76 (approximately 35–40%), a high pressure inside the extraction
3 78 73 75 vessel (approximately 25–30 psi) and a moderate extraction
4 70 69 87 time (approximately 15–25 min). Nevertheless, these 3D plots
5 69 71 80
do not consider the variation of the third variable, so the
common optimum conditions for the five phenols might be
estimated by the simplex method implemented in the Multi-
Simplex program. For this purpose, the parameters given in
Table 6 Relative standard deviations for the five phenols using MAE Table 3 were introduced into an Excel spreadsheet, where the
extraction recoveries for each of the phenols were calculated
RSD (%) under each set of experimental conditions proposed by
Within Among
MultiSimplex. Thus, it was possible to obtain the ranges in
Compound Analysis days days Total which the variables should change in order to obtain a recovery
as close to 100% as possible. Eventually, despite the simulation
Phenol 3.03 4.84 4.84 7.48
2-Chlorophenol 2.16 5.40 6.76 8.92
of the extraction for a high number of experiments, only phenol,
2-Methylphenol 1.97 4.08 10.20 10.99 2-nitrophenol and 2,4-dichlorophenol could reach approx-
2-Nitrophenol 4.18 30.00 18.89 30.80 imately 100% recovery. For the rest of the phenols recoveries of
2,4-Dichlorophenol 1.88 12.50 7.69 14.80 approximately 88% (2-chlorophenol) and 75% (2-methylphe-
nol) were obtained. These recoveries were obtained for a
percentage of microwave power of 44%, a time of extraction of
16.5 min and a pressure inside the extraction vessel of 26 psi.
Table 7 Relative standard deviations for the five phenols using Soxhlet
extraction
Complete factorial design for Soxhlet extraction
RSD (%)
The variables considered in the complete factorial design for
Within Soxhlet extraction were the percentage of acetone in the
Compound Analysis days Total acetone–hexane mixture and the extraction time. A two-level
Phenol 2.45 17.06 17.23 complete factorial design involving four runs was chosen. The
2-Chlorophenol 2.32 0.85 2.47 design matrix and the results for the six runs (the first run of the
2-Methylphenol 10.47 7.82 13.07 experiment was carried out in triplicate) are given in Table 4.
2-Nitrophenol 20.91 21.16 29.75 Samples were measured by GC–FID and losses that occurred
2,4-Dichlorophenol 1.91 8.96 9.16 during evaporation were corrected.
The most general function allowed for the complete factorial
design carried out is
Y = b0 + b1x1 + b2x2 + b12x1x2 (7)
where x1 is extraction time and x2 is percentage (v/v) of acetone
in the acetone–hexane mixture. The results obtained by NLREG
did not show a similar trend that could be observed (sign or
magnitude) among the parameters calculated. For this reason,
the optimum conditions were fixed taking into account the
experimental data in Table 4. It could be observed that
increasing the percentage (v/v) of acetone in the solvent gave
better recoveries but an increase in the extraction time did not
yield much higher recoveries. Hence optimum conditions for
the extraction of the five phenols in soils by Soxhlet extraction
Fig. 3 Recoveries of the five phenols under optimum conditions for both were 200 ml of a acetone–hexane mixture (70 + 30 v/v) under
MAE and Soxhlet extraction methods. reflux for 8 h.
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1684 Analyst, August 1998, Vol. 123

Evaluation of reproducibility for MAE shows the recoveries for the five phenols under the optimum
conditions for both the MAE and Soxhlet extraction methods. It
For evaluating the reproducibility of the measurements for
can be concluded that for phenol, 2-chlorophenol and 2,4-di-
MAE, five aliquots of the sample were extracted each day and
chlorophenol comparable results were obtained by both meth-
this procedure was repeated three times under the optimum
ods. For 2-methylphenol and 2-nitrophenol better results were
conditions mentioned above [15 ml of acetone–hexane (1 + 1
obtained by MAE. In both methods, the recoveries obtained for
v/v) at 26 psi with a microwave power of 44% for 16.5 min). In
2-methylphenol were low, but other workers have reported low
other studies,7–11 the reproducibility of MAE among days was
recoveries for 2-methylphenol18 and other alkylphenols.19
not studied. The results obtained for 2-chlorophenol during the
These low recoveries are attributed to interactions between the
three days are given in Table 5.
alkyl groups and the soil matrix. The relative standard
The results were analysed by means of analysis of variance
deviations for MAE are higher than those obtained for Soxhlet
(ANOVA) of the set of experimental data in Table 5. It could be
extraction but it must be taken into account that the variance
observed, for a degree of confidence of 95%, that there were no
among days was considered for MAE but not for Soxhlet
significant differences among samples extracted within a day
extraction.
(Fexp = 0.18 < Fcrit = 3.84), but there were significant
differences for samples extracted among days (Fexp = 5.47 >
This research was supported by the Basque Government
Fcrit = 4.46). This behaviour was similar for phenol, 2-chloro-
Published on 01 January 1998. Downloaded on 24/10/2014 17:58:37.

(project PIGV96/74). Olatz Zuloaga is grateful for a Scholar-

phenol and 2-methylphenol. 2-Nitrophenol showed significant
ship granted by the Basque Government.
differences for samples extracted within a day but not among
days (the RSDs were too large and all values among days were
not significantly different) and 2,4-dichlorophenol showed no References
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Sci., 1995, 33, 658.
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3 Llompart, M., Lorenz, R., and Cela, R., J. High Resolut. Chroma-
variance due to the analysis, the variance within days and the togr., 1996, 19, 207.
variance among days as indicated in the equation 4 Bosch, F., Campins, P., and Verdu, J., J. Liq. Chromatogr., 1995, 18,
stot2 = sa2 + sw,d2 + sa,d2 (8) 2229.
5 Heemken, O. P., Norbert, T., and Wenclawiak, B. W., Anal. Chem.,
where stot2 is the total variance, sa2 is the variance due to the 1997, 69, 2171.
analysis (which was calculated considering error propagation 6 Snyder, J., Grob, R., McNally, M., and Oostdyk, T., Anal. Chem.,
and making use of the spreadsheet method17), sw,d2 is the 1992, 64, 1940.
variance within days and sa,d2 is the variance among days. 7 Barnabas, I. J., Dean, J. R., Fowlis, I. A., and Owen, S. P., Analyst,
1995, 120, 1897.
Table 6 gives the relative standard deviations for the analysis,
8 Llompart, M. P., Lorenzo, R. A., Cela, R., and Paré, J. R. J., Analyst,
for samples within a day, samples among days and total relative 1997, 122, 133.
standard deviations estimated from eqn. (8) The RSD values 9 Chee, K. K., Wong, M. K., and Lee H. K., J. Chromatogr. A, 1996,
were good for all phenols except 2-nitrophenol. 723, 259.
10 Onuska, F. I., and Terry, K. A., J. High Resolut. Chromatogr., 1995,
18, 417.
Evaluation of reproducibility for Soxhlet extraction 11 Pastor, A., Vázquez, E., Ciscar, R., and de la Guardia, M., Anal.
Chim. Acta, 1997, 344, 241.
For studying the reproducibility of Soxhlet extraction, three 12 Nelder, J. A., and Mead, R., Comput. J., 1965, 7, 308.
samples were extracted under the same conditions (run No. 1 in 13 Bergström, and Öberg, MultiSimplex (1.0 Version), Gullberna Park,
the experimental design, Table 4). The variance associated with Karlskrona, 1997.
each measure was expressed as in eqn. (8). In this case the 14 ISO/DIS 11464, Soil Quality–Pretreatment of Samples for Physico-
variance among days was not considered in order to estimate the chemical Analyses, US EPA, Washington, DC, 1991.
total variance of the results. Table 7 gives the relative standard 15 Kateman, G., and Buydens, L., Quality Control in Analytical
deviations corresponding to the analysis and samples extracted Chemistry, Wiley, New York, 2nd edn., 1993.
within a day for Soxhlet extraction. 16 Sherrod, P. H., NLREG—Nonlinear Regression Analysis Program,
Nashville, TN, 1995.
17 Kragten, J., Analyst, 1994, 119, 2161.
Comparison of Soxhlet and MAE methods 18 Lopez-Avila, V., and Young, R., Anal. Chem., 1994, 66, 1097.
19 Dean, J. R., Santamaria-Rekondo, A., and Ludkin, E., Anal.
The mean values for the extraction carried out by MAE and Commun., 1996, 33, 413.
Soxhlet extraction under the optimum conditions [15 ml of
acetone–hexane (1 + 1 v/v) at 26 psi with a microwave power of Paper 8/02117A
44% and for 16.5 min and 200 ml of acetone–hexane (70 + 30 Received March 11, 1998
v/v) under reflux for 8 h, respectively] were compared. Fig. 3 Accepted May 29, 1998