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© 2012 American Chemical Society 5816 dx.doi.org/10.1021/ac301274r | Anal. Chem. 2012, 84, 5816−5822
Analytical Chemistry Article
Soxhlet extraction, developed in 1879, has been the most Ginsengs were collected from America (Wisconsin), Canada
widely used extraction technique for isolating target analytes (Ontario), and Northeast China (Jilin). After being dried in a
from solid samples.18 To improve Soxhlet performance, a hot air cabinet with ventilation at 40 °C for 5 h, the ginsengs
variety of modifications have been made, and the most success were triturated, passed through a stainless steel sieve with 0.45
was the use of microwaves as an auxiliary energy.19,20 mm mesh sizes and stored in closed desiccators. The same
Microwave-assisted Soxhlet extraction (MA-SE), developed by batch of samples was used here in the experiments.
Luque De Castro et al.,21 is an approach based on the same Apparatus. MA-SSE experiments were performed on an
principles as conventional Soxhlet extraction but using MAS-II microwave extraction apparatus (Sineo Microwave
microwaves to accelerate the process. Since 1998, three Chemistry Technology Company, Shanghai, China). Micro-
different prototypes of MA-SE have been designed and wave irradiation, with a maximum delivered power of 1000 W
constructed. The first prototype was constructed by Prolabo at a frequency of 2450 MHz, can be dynamically adjusted by
(Paris, France). Solvent distillation in this extractor is achieved temperature and power feedbacks, which is able to provide
by electrical heating, which is independent of the extractant continuous nonpulse microwave heating. Temperature is
polarity.22 The main drawback of the first extractor is the monitored by an infrared sensor installed on the inner wall.
difficulty of using high-boiling-point solvents.23 The second GC/MS analysis was carried out on a Shimadzu GC-MS
prototype was constructed by SEV (Puebla, Mexico), called QP2010 system (Shimadzu, Kyoto, Japan). A DB-5MS
MIC II. By shortening the distillation glassware, this prototype (Agilent, Palo Alto, USA) capillary column (30 m long ×
has the availability to work with high-boiling-point solvents.24 0.25 mm i.d. × 0.25 μm film thickness) was used for the
Finally, a fully automated MA-SE extractor was constructed by chromatographic separation. The chromatographic data were
SEV. This extractor named, MIC V, operates with two recorded and processed with GC/MS solution software
extraction units, allowing the simultaneous processing of two (version 2.10) supplied by Shimadzu.
samples.25 In addition to the extraction of lipids from food and SEM images were conducted by a Quanta 600 scanning
other products, MA-SE has been widely applied to the analysis electron microscopy (SEM) instrument (FEI, Hillsboro, OR,
of organochlorine pesticides,26 polycyclic aromatic hydro- USA).
carbons,25 and polychlorinated biphenyls27 in natural products Sorbent Conditioning. Florisil was heated in a muffle
and environmental samples. Although MA-SE has been proved furnace at 550 °C for 6 h and cooled in a desiccator, then was
to be rapid and efficient, its low selectivity often leads to further added with purified water (equivalent to 5% w/w) and
cleanup steps.26,28 The processes are relatively laborious and homogenized by rotation for 2 h. After that, the Florisil was
time-consuming. Therefore, a selective MA-SE technique is left in a closed container to equilibrate for 48 h before use.
needed. Diatomaceous earth was boiled in a 5% HCL aqueous solution
In view of this, a novel one-step sample preparation for 1 h and then was filtered and washed with water until the
technique called microwave accelerated selective Soxhlet pH value was 6.0. After that, diatomaceous earth was heated in
extraction (MA-SSE) was developed in this study. During the a muffle furnace at 500 °C for 2 h. The silicone was heated at
extraction procedure, the analytes and interfering compounds 110 °C for 1 h and cooled in a desiccator. Neutral alumina was
of samples were extracted simultaneously into the extraction heated at 450 °C for 4 h and cooled in a desiccator.
solvent. When the solvent flowed though the sorbent, the Extraction Procedures. Microwave Accelerated Selective
interfering components were retained by the sorbent while the Soxhlet Extraction. A 1.0 g portion of ginseng sample was
analytes remained in the extraction solvent. The whole accurately weighed and put into the Soxhlet thimble after the
procedure was accelerated by microwave irradiation, and no addition of 2.0 g of anhydrous sodium sulfate and a certain
cleanup or filtration preceded chromatographic analysis. To amount of sorbent. The entire filler was then compressed to
demonstrate the feasibility of the approach, MA-SSE was form a column packing. A certain volume of extraction solvent
applied to analysis of organophosphorus pesticide (OPP) and and a stirrer were added into the 50 mL round-bottom flask.
carbamate pesticide residues in ginseng with gas chromatog- MA-SSE was performed at a certain temperature with a certain
raphy/mass spectrometry (GC/MS). The mechanism of MA- power of microwave irradiation. The extraction conditions were
SSE was investigated systematically, including the effect of optimized by a monofactor test. The extract obtained was
microwave irradiation, the cleanup ability of Florisil, and the concentrated to 1.0 mL under a nitrogen stream in a 40 °C
microstructure analysis. The mass transfer procedure was finally water bath.
proposed. Selective Soxhlet Extraction. A 1.0 g portion of ginseng
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sample was accurately weighed and put into the Soxhlet thimble
after the addition of 2.0 g of anhydrous sodium sulfate and 3.0 g
EXPERIMENTAL SECTION of Florisil. The entire filler was then compressed to form a
Reagents and Samples. Standards of OPP and carbamate column packing, then 30 mL of ethyl acetate and a stirrer were
pesticides were purchased from The Agro-Environment added into the 50 mL round-bottom flask. The experiment was
Protection Institute (Tianjin, China). All standard solutions performed at 85 °C in a thermostatic water bath for some time,
were prepared in acetone and were stored at −20 °C. Florisil and the stirring speed was 1000 rpm. The extract obtained was
with 60−100 mesh was purchased from Supelco (Bellefonte, concentrated to 1.0 mL under a nitrogen stream in a 40 °C
PA, USA). Neutral alumina, silicone, and diatomaceous earth, water bath.
all of which were the same size of 60−100 mesh, were Microwave-Assisted Soxhlet Extraction. A 1.0 g portion of
purchased from Sinopharm Chemical Reagent Co., Ltd. ginseng sample was accurately weighed and put into the Soxhlet
(Shanghai, China). Ethyl acetate, acetone, dichloromethane, thimble after the addition of 2.0 g of anhydrous sodium sulfate.
and acetonitrile were of analytical grade and were purchased The entire filler was then compressed to form a column
from Guangzhou Chemical Reagents Factory (Guangzhou, packing, then 30 mL of ethyl acetate and a stirrer were added
China). Distilled water was used throughout the study. into the 50 mL round-bottom flask. The experiment was
5817 dx.doi.org/10.1021/ac301274r | Anal. Chem. 2012, 84, 5816−5822
Analytical Chemistry Article
■
alumina, underivatized silica, and diatomaceous earth. The
results showed that the cleanup abilities of the sorbents
RESULTS AND DISCUSSION decreased in the following order: Florisil > diatomaceous earth
Actualization of Microwave Accelerated Selective > underivatized silica > neutral alumina. The cleanup abilities of
Soxhlet Extraction. MA-SSE was performed with an underivatized silica and neutral alumina were so poor that the
apparatus shown in Figure 1. It consists of a microwave analytes were severely interfered during GC/MS analysis. The
extraction device, a reflux condenser, and an improved Soxhlet adsorption of diatomaceous earth is nonselective, which
extractor. The improved Soxhlet extractor comprises a Soxhlet resulted in low recoveries. Therefore, Florisil was chosen as
thimble and a round-bottom flask. The Soxhlet thimble the sorbent for further application on the basis of the best
containing a glass filter with 10−15 μm pore size is used to compromise between cleanup ability and extraction efficiency.
hold the entire sample and sorbent. The extraction solvent is The extraction solvents tested in this study were ethyl
added into the round-bottom flask. The cooling system fitted acetate, acetone, dichloromethane, and acetonitrile. Figure S-1A
on top of the microwave cavity refrigerates the solvent of the Supporting Information shows the influence of extraction
continuously, and the solvent is refluxed into the Soxhlet solvent on the extraction efficiency. The results demonstrated
extractor. that ethyl acetate and acetone obtained higher extraction yields
Before the extraction, the sorbent and sample are added into than dichloromethane and acetonitrile. That is because OPP
the Soxhlet thimble in order, and the extraction solvent is and carbamate pesticides are polar molecules; thus, the
added into the round-bottom flask. During the extraction extraction effect of less polar solvents, such as dichloromethane,
procedure, both the extraction solvent and sample are heated is poor based on the principle of the dissolution in the similar
by the microwave irradiation. When the solvent begins to reflux, material structure. In this case, ethyl acetate, which is less toxic
both the target analytes and the interfering compounds in the than acetone, was chosen as the extraction solvent for further
sample are extracted into the extraction solvent. After the application.
5818 dx.doi.org/10.1021/ac301274r | Anal. Chem. 2012, 84, 5816−5822
Analytical Chemistry Article
Table 1. Analytical Performance of GC/MS for the Determination of OPP and Carbamate Pesticides
no. pesticides tR (min) equation linear range (μg/kg) R LOD (μg/kg)
1 isoprocarb 8.36 Y = 2.61 × 102 X − 2.35 × 10 0.50−400 0.9998 0.10
2 phorate 9.64 Y = 4.26 × 102 X − 9.24 × 102 0.50−400 0.9996 0.050
3 iprobenfos 11.25 Y = 1.92 × 102 X + 9.96 × 10 0.50−400 0.9998 0.10
4 chlorpyrifos methyl 11.81 Y = 2.04 × 102 X + 1.30 × 102 0.50−400 0.9996 0.10
5 parathion methyl 12.00 Y = 8.58 × 10 X + 5.13 × 10 1.0−400 0.9993 0.20
6 tolclofos methyl 12.02 Y = 3.15 × 102 X + 2.61 × 102 0.50−400 0.9996 0.10
7 malathion 13.06 Y = 1.18 × 102 X − 2.97 × 102 2.0−400 0.9996 0.50
8 chlorpyrifos 13.28 Y = 6.09 × 10 X + 1.57 × 102 0.50−400 0.9999 0.10
Figure 6. Extraction process of the MA-SSE technique: (A) before extraction, (B) during extraction, and (C) after extraction.
There were many more interferences in the extract obtained by surface of the ginseng shrank a little after SSE, probably because
MA-SE, which seriously interfered with the analysis of of the high extraction temperature and the long extraction
malathion and chlorpyrifos. time.18 The surface of the ginseng after UAE was still intact
As a normal phase sorbent, Florisil can adsorb strongly polar without any ruptures. The results indicate that in the SSE and
compounds, such as pigments and other interfering compo- UAE process, the solvent transferred into the matrix and
nents from sample matrixes.35 The retention of the interfering extracted the target compounds by solubilization. Therefore, a
compounds by Florisil is primarily due to the interactions long extraction time was needed in SSE and low recoveries
between polar functional groups such as hydrogen bonding.36 were obtained by UAE.
Moreover, Florisil is a nice fat retainer and could give fat-free The Mass Transfer and Prospect of MA-SSE. The mass
extracts of ginseng.8 According to the results, Florisil can transfer of MA-SSE was proposed on the basis of the above
adsorb the major interferences in ginseng effectively, and the results, which is shown in Figure 6. MA-SSE was divided into
use of Florisil would prevent the GC system from serious three procedures. Before the extraction, the ginseng samples
pollution. were placed on the sorbents. During the extraction procedure
Microstructure Analysis. To further investigate the mecha- of MA-SSE, the extraction solvent was heated up to the boiling
nism of MA-SSE, SEM was used to identify the microstructures point and the steam was generated. After the condensation, the
of samples over various conditions, which are shown in Figure solvent drips flowed though the samples and the sorbents.
S-2 of the Supporting Information. Distinguishable physical Meanwhile, the cell walls of the ginseng samples were ruptured
changes produced by different extraction methods were under the microwave irradiation, and the inside target analytes
observed. and interfering compounds dissolved and dispersed into the
Compared with the intact surface of the untreated ginseng extraction solvent simultaneously. When the extraction solution
sample (Figure S-2A), the surface of the sample after MA-SSE flowed though the sorbent, the target analytes and interfering
(Figure S-2B) was greatly destroyed, and the cell walls were compounds distributed between the extraction solvent and the
ruptured. The result was in accord with the hypothesis of MAE sorbent on the basis of their interactions. The normal phase
proposed by Paré et al.37 The rapid increase in the temperature sorbent Florisil can adsorb strongly polar interfering com-
generated by microwave heating caused the rupture of the cell pounds and most lipids in the samples.35 Ethyl acetate, the
walls, leading to the quick migration of the compounds in the extraction solvent used in this study, has enough polity to elute
cells into the extraction solvent.38 In contrast, the micro- the pesticides but could not elute more polar interfering
structures of the ginseng samples after SSE and UAE (Figure S- compounds. Thus, after the extraction, the interfering
2C and S-2D, respectively) were not significantly changed. The compounds were retained by the sorbent, and the pesticides
5821 dx.doi.org/10.1021/ac301274r | Anal. Chem. 2012, 84, 5816−5822
Analytical Chemistry Article
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polar target analytes in complex matrixes. This technique will
(23) García-Ayuso, L. E.; Luque De Castro, M. D. Trends Anal. Chem.
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*
ASSOCIATED CONTENT
S Supporting Information
Chromatogr., A 2003, 994, 159−167.
(26) Prados-Rosales, R. C.; García, J.; de Castro, M. J. Chromatogr., A
2003, 993, 121−129.
(27) Luque-García, J. L.; de Castro, M. J. Chromatogr., A 2003, 998,
Additional information as noted in text. This material is 21−29.
available free of charge via the Internet at http://pubs.acs.org.
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(28) Morales-Muñoz, S.; Luque-García, J. L.; Ramos, M. J.; Martínez-
Bueno, M. J.; Luque De Castro, M. D. Chromatographia 2005, 62, 69−
AUTHOR INFORMATION 74.
Corresponding Author (29) Kristenson, E. M.; Brinkman, U. A. T.; Ramos, L. Trends Anal.
*Phone: +86-20-84110922. Fax: +86-20-84115107. E-mail: Chem. 2006, 25, 96−111.
(30) Hajšlová, J.; Zrostlíková, J. J. Chromatogr., A 2003, 1000, 181−
(G.L.) cesgkl@mail.sysu.edu.cn, (X.X.) xiaoxhua@mail.sysu. 197.
edu.cn. (31) González-Rodríguez, R. M.; Rial-Otero, R.; Cancho-Grande, B.;
Notes Simal-Gándara, J. J. Chromatogr., A 2008, 1196−1197, 100−109.
The authors declare no competing financial interest. (32) Luque-García, J. L.; Luque De Castro, M. D. Trends Anal. Chem.
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2003, 22, 41−47.
ACKNOWLEDGMENTS (33) Pérez-Serradilla, J. A.; Priego-Capote, F.; Luque De Castro, M.
D. Anal. Chem. 2007, 79, 6767−6774.
This work was supported by the Natural Science Foundation of (34) Sparr Eskilsson, C.; Björklund, E. J. Chromatogr., A 2000, 902,
China (No. 21127008 and 20905080) and the Guangdong 227−250.
Provincial Department of Science and Technology of China (35) Moreda-Piñeiro, J.; Alonso-Rodríguez, E.; López-Mahía, P.;
(No. 2009B010900021) and the Fundamental Research Funds Muniategui-Lorenzo, S.; Prada-Rodríguez, D.; Romarís-Hortas, V.;
for the Central Universities (No. 11lgpy80). Míguez-Framil, M.; Moreda-Piñeiro, A.; Bermejo-Barrera, P. Trends
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