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1. Introduction
During the World War II, Percy Spencer discovered the heating
properties of microwaves while experimenting with radar. Actually,
with a candy bar in his pocket, he leaned in front of the microwave
tube and the candy bar promptly melted. Thus, the heating property of microwaves was evident and this event led to the birth of
microwave ovens after the World War II.
Microwaves are nonionizing electromagnetic waves of frequency (electromagnetic radiations) with a frequency from 0.3 to
Satyajit D. Sarker and Lutfun Nahar (eds.), Natural Products Isolation, Methods in Molecular Biology, vol. 864,
DOI 10.1007/978-1-61779-624-1_5, Springer Science+Business Media, LLC 2012
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Table 1
Dielectric constant and dissipation factor of some common
solvents used in extraction process
Solvents
Ethanol (EtOH)
1.96
2,500
Methanol (MeOH)
2.87
6,400
2-Propanol
1.66
6,700
Water
2.3
1,570
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2. Materials
2.1. Microwave
Extractors
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Extraction vessels
Diffused microwaves
Turntable
Water outlet
Reflux system
Water inlet
Magnetron
Wave guide
Extraction
vessel
Solvent
Focussed
microwaves
Sample
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Extraction tank
Outlet
Microwave
Microwave
Microwave
Refrigerant
Valve
Inlet
Pump
Controller
Container
Magnetic stirrer
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Table 2
Manufacturers/suppliers of microwave extraction apparatus
Model
Hangzhou Nade
http://zjnade.en.alibaba.com/
Scientific Instrument
Co., Ltd.
ETHOS One
Advanced Microwave
Extraction Labstation
Milestone
http://www.milestonesrl.com/analytical/
products-microwave-extraction-ethos-one.
html
Anton-Paar
Microwave extraction
machine for roots
Wenzhou Chengdong
Import & Export
Co., Ltd.
http://wzchengdong.en.alibaba.com/
Ultrasonic-microwave
extractor and reactor
Lab-Kits Lab
Instruments
and Facilities
2.2. Solvents
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3. Methods
In the case of MAE, the extraction process is based on exposing
the analytes to the solvent through cell rupture. Desorption of
components from plant matrix takes place in MAE. The free
water molecules present in the gland and vascular systems of plant
matrices are heated and this results in localized heating and
expansion of their walls. As a result, plant constituents flow outside the cells. The dielectric susceptibility of the solvent and
matrix is the factor that affects utilization of microwave energy in
the process. Use of additional extraction solvents increases the
efficiency of this extraction method. In order to avoid degradation
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3.2.1. Extraction
of Essential Oils by MAE
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5. A mixture of hot crude juice and steam (in situ water) should
move downward by gravity into a spiral condenser outside the
microwave cavity where it is condensed.
6. Collect the oily condensate is collected continuously in a
receiving flask (similar to separator funnel).
7. Essential oil will form a film on the surface of the water. Skim
the film off the top and collect the essential oil.
8. Dry collected essential oil with anhydrous sodium sulfate and
store at 4C until used.
3.2.2. Extraction
of Flavonoids
and Isoflavonoids by MAE
Zhang et al. (24) reported MAE of rutin and quercetin from the
stalks of Euonymus alatus (Thunb.) Sieb. and compared with conventional extraction techniques. MAE was performed with an
MSP-100E multimode microwave extraction system (maximal
power: 850 W) in the optimized conditions of 50% EtOH (v/v)
solution, 40 mL the extractant volume, 170 W microwave power,
and 6 min irradiation time. MAE was found to be the fastest of all
extraction methods compared, requiring just 6 min, with a higher
yield and lower solvent consumption for extraction of rutin and
quercetin in plants
An approach for automated, continuous and rapid extraction
of flavonoids from the dried cell cultures of Saussurea medusa
Maxim., utilizing a fully optimized dynamic microwave-assisted
extraction (DMAE) system, was described (25). The yield of flavonoids reached ~5% in 60 min under the optimum extraction
conditions: 1,200 W of radiation power, 50:1 (v/w) of the liquid/
solid ratio, and 50 mL/s of solvent flow rate. The dynamic MAE
displayed distinct advantages in short duration and high efficiency
to extract flavonoids without causing degradation of target components from the S. medusa dried cell cultures in comparison with the
dynamic solvent extraction without microwave assistance. The
protocol of this extraction can be outlined as follows:
1. Use a dynamic MAE system as outlined in Fig. 3.
2. Place a given amount of S. medusa dried cell cultures and 2.0 L
of 80% EtOH into the container, and mix homogeneously by
a magnetic stirrer.
3. Recycle the mixed extraction sample between the container
and three extraction tanks by the liquid bump.
4. Irradiate the suspension with 60 s of power ON to reach the
desired temperature of about 80C, and then irradiate periodically with microwaves in a presetting procedure (15 s of power
ON for heating followed by 15 s of power OFF for cooling
without allowing the suspension to super-boil).
5. Carry out qualitative and quantitative analyses of flavonoids by
HPLC.
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Pan et al. (47) extracted polyphenols as well as the xanthine alkaloid, caffeine, from green tea (Thea sinensis L.) leaves utilizing the
MAE method resulting in good extraction efficiency. A household
microwave oven was used in evaluating different experimental conditions to optimize the extraction. The extraction efficiency of
polyphenols and caffeine was increased with the increase of MAE
time and the pre-leaching time from 4 to 90 min, as well as the
increase in liquid/solid ratio. Among the various solvents examined for the extraction, acetone and MeOH obtained higher extraction values for polyphenols and caffeine, respectively.
Ionic liquid (see Note 6)-based microwave-assisted extraction
(ILMAE) has recently been developed for the extraction of three
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R
N
R
Alkaloids
N-Nornuciferine
OMe
OMe
O-Nornuciferine
OH
OMe
Me
OMe
OMe
Me
Nuciferine
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different anions, e.g., Cl, Br, BF4, and PF6, the most efficient
one was Br, which demonstrated that the extraction efficiency of
alkaloids was anion-dependent (see Note 7). This study also evaluated the effect of alkyl side chain; with the same anion of Br, a
series of 1-alkyl-3-methylimidazolium cations, including C2mim+,
C4mim+, C6mim+, and C8mim+ were studied. It was observed that
the extraction efficiency increased significantly when the alkyl chain
length was increased from ethyl to hexyl. However, increasing alkyl
chain length from hexyl to octyl dramatically decreased the extraction efficiency (see Note 8). A similar protocol was also reported
for the extraction of alkaloids from Nelumbo nucifera (49).
The use of the nonionic surfactant 5% oligoethylene glycol
monoalkyl ether (Genapol X-080) as a solvent for a focused MAE
of alkaloids from Rhizoma Coptidis was found to enhance the
extraction efficiency significantly with a massive 92.8% recovery of
alkaloids in a one-step and fully optimized extraction (50).
Focused MAE of cocaine and benzoylecgonine from the leaves
of Erythroxylum coca var. coca was reported by Brachet et al. (51)
utilizing a Soxwave (Prolabo, France) model 3.6 at atmospheric
pressure and a frequency of 2,450 MHz. Different solvents, particle size, radiation time, and power were evaluated to study their
effects and optimize the extraction condition. Because of the ability of MeOH for absorbing microwave energy and solubilizing
cocaine, it was found to be the best extraction solvent. On the
contrary, no quantitative extraction of cocaine was established with
nonpolar solvents having low dielectric constants (hexane and toluene). Smaller particle size distribution provided an enhanced
extraction of cocaine, revealing an evident influence of particle size
on the extraction efficiency. Radiation time and the applied power
had no obvious effect on cocaine recovery. Under optimum conditions of 90150 m particle size, 5 mL of MeOH, 30 s of radiation
time and microwave power of 125 W, the cocaine content was
determined as 0.6800.004% by GCFID analysis and 0.100.02%
for benzoylecgonine.
3.2.6. Extraction
of Saponins by MAE
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3.2.7. Extraction
of Endogenous Steroids
in Marine Mussels Mytilus
sp. (58)
3.2.8. Extraction
of Chlorogenic Acid
from Lonicera japonica
by MAE (59)
3.2.9. Extraction
of Paclitaxel from Taxus
baccata by MAE (60)
4. Notes
1. Ionic conduction refers to the electrophoretic migration of
ions under the influence of the changing electric field.
2. The chosen power has to be set correctly to avoid excessive
temperatures leading to possible solute degradation and overpressure problems.
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