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Herbal Processing and Extraction Technologies

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ISSN: 1542-2119 (Print) 1542-2127 (Online) Journal homepage: http://www.tandfonline.com/loi/lspr20

Herbal Processing and Extraction Technologies

Siti Nuurul Huda Mohammad Azmin, Zainuddin Abdul Manan, Sharifah

Rafidah Wan Alwi, Lee Suan Chua, Azizul Azri Mustaffa & Nor Alafiza Yunus

To cite this article: Siti Nuurul Huda Mohammad Azmin, Zainuddin Abdul Manan, Sharifah
Rafidah Wan Alwi, Lee Suan Chua, Azizul Azri Mustaffa & Nor Alafiza Yunus (2016) Herbal
Processing and Extraction Technologies, Separation & Purification Reviews, 45:4, 305-320, DOI:
10.1080/15422119.2016.1145395

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Separation & Purification Reviews, 45: 305–320, 2016
Copyright © Taylor & Francis Group, LLC
ISSN: 1542-2119 print / 1542-2127 online
DOI: 10.1080/15422119.2016.1145395

Herbal Processing and Extraction Technologies

Siti Nuurul Huda Mohammad Azmin,1,2 Zainuddin Abdul Manan,1,2 Sharifah
Rafidah Wan Alwi,1,2 Lee Suan Chua,3 Azizul Azri Mustaffa,1,2
and Nor Alafiza Yunus1,2
1
Process Systems Engineering Centre (PROSPECT) Research Institute of Sustainable Environment
(RISE), Universiti Teknologi Malaysia, UTM Johor Bahru, Johor, Malaysia
2
Faculty of Chemical and Energy Engineering, Universiti Teknologi Malaysia, UTM Johor Bahru,
Johor, Malaysia
3
Institute of Bioproduct Development (IBD), Universiti Teknologi Malaysia, Johor Bahru, Johor,
Malaysia

Herbs are widely utilized in food and health industries. Their beneficial effects to the human
body have been attributed to the presence of active phytochemical ingredients with some effi-
ciency for disease treatment as well as for beauty and health enhancement. Public awareness on
the adverse effects of synthetic chemical products also increased the demand for herbal prod-
ucts. Highly efficient herbal processing and extraction technologies have been developed to
obtain the optimal amounts of active ingredients from herbs and cope with the rising demands
for herbal products. This article reviews the state-of-the-art development in herbal processing
and extraction methods from the year 1991 until 2015. We start with a brief history of herbal
usage, followed by descriptions of 10 types of extraction processes and critical analysis of
their relative advantages and disadvantages. Scale-up considerations of the extraction methods
are shared, and a highlight of the current and future challenges facing the herbal industry is
presented.

Keywords: Extraction technologies, herbs, herbal processing, natural products,

phytochemicals

INTRODUCTION or in combination, to treat, diagnose and prevent illnesses

Throughout human history, herbs have been used in food,
cosmetics and fragrances (1). In addition, they are a source
of traditional medicines for the treatment of various ill-
nesses and diseases (2). Traditional medicine has been
defined by the World Health Organization (WHO) as “health
practices, approaches, knowledge and beliefs incorporating
plant, animal and mineral-based medicines, spiritual ther-
apies, manual techniques and exercises, applied singularly
Received 30 December 2013, Accepted 14 January 2016.
Address correspondence to Sharifah Rafidah Wan Alwi, Process
Systems Engineering Centre (PROSPECT), Research Institute of
Sustainable Environment (RISE), Universiti Teknologi Malaysia, UTM,
Johor Bahru, Johor, Malaysia. E-mail: syarifah@utm.my
Color versions of one or more of the figures in the article can be found
online at www.tandfonline.com/lspr.
or maintain well-being” (3). Herbal products have recently
increased in importance in the treatment of health-related
problems. A recent WHO report (3) stated that 5.6 billion
people, or approximately 80% of the world’s population, use
herbal products for their primary health care (3–5). Herbal
products are currently being used as dietary supplements,
nutraceuticals, health products, and traditional medicines.
These products also are being prescribed by Chinese and
Indian national pharmacopeias (5).
The botanical dietary supplement market in the United
States has expanded rapidly from US$ 2.9 billion in 1995 (6)
to US$ 4.8 billion in 2008 (7). Over 42% of the U.S popula-
tion has claimed to have used botanical dietary supplements
either to widen their current diet or to treat or to prevent
health-related issues (5). European dietary supplements
have contributed to a steady 4% growth rate of the market
between 2004 and 2011. The total market in 2004 was
306 S. N. H. M. AZMIN ET AL.
valued at US$ 6.2 billion (8). With Malaysians consuming
over US$ 1.44 billion worth of herbal-related products
(with over 80% imported) (9), the opportunity to increase
herbal product processing in Malaysia has expanded. From
2000 to 2005, annual sales of traditional medicines increased
from US$ 385 million (RM1 billion) to US$ 1.29 billion
(RM45 billion), as reported by Malaysian National News
Agency, BERNAMA (10). The expected growth of the
herbal medicine industry is between 15% and 20% per year
(11).
With the increased demand for herbal products, Malaysia
is well-positioned to be a key global player in this field
because of its rich biological resources, cultural background,
and trade links (1). In addition, Malaysia is ranked fourth in
Asia and twelfth in the world as the most biodiverse nation,
with 3,000 species of medicinal plants and 15,000 species
of flowering plants (12). Among the 3,000 known medic-
inal plants (13), the top Malaysian traditional medici-
nal plants are Tongkat Ali (Eurycoma longifolia), Kacip
Fatimah (Labisia pumila), Hempedu Bumi (Andrographis
paniculata), Dukung Anak (Phyllantus amarus), Misai
Kucing (Orthosiphon stamineus), and Pegaga (Centella asi-
atica). Among these known plants, less than 50 traditional
medicine species have been scientifically investigated, with
the remainder not yet characterized (1). Worldwide, more
than 35,000 plant species have been reported to be employed
for medical purposes (14). Even with this extensive number
of available plant species, Rates (15) has reported that only
121 active compounds from herbs have been utilized, with
approximately 25% of prescribed drugs originating from
these plants.
Medicinal plants have often been applied for a broad
range of acute and chronic conditions (16). This provides
ample opportunity to gain a vast wealth of knowledge
through exploration of these plants. Phytochemical con-
stituents in medicinal plants, such as flavonoids, phenolics,
and saponins (17), are believed to have numerous thera-
peutic abilities and are able to reduce the risk of multiple
diseases, including inflammatory conditions and cancer (18).
They contribute to the protection of herbs from microbial
and insect invasions (19). Flavonoids have been utilized to
improve human health via their multiple biological functions
including anti-inflammatory (20), antimicrobial (21), antiox-
idant (22), and anticancer activities (23) and the prevention
of osteoporosis (24).
Tsao and Deng (25) have reported that phenolic acids
from herbs have antioxidant activities that exceed the val-
ues exhibited by vitamins C and E. Polyphenols have also
been applied in the prevention of both cancer and heart dis-
ease (19). Aqueous extracts of the Malaysian herb L. pumila,
commonly known as Kacip Fatimah, have been demon-
strated to protect significantly human dermal fibroblasts from
cell damage caused by ultraviolet irradiation (26), most
likely as a result of the presence of flavonoids (27). Ginseng,
one of the most commonly used herbs, contains saponins as
active ingredients (28). Ginseng exhibits a broad range of
biological properties, including aphrodisiac and adaptogenic
activities (29).
With the high demand for herbal products that promote
health advantages, the identification of the most effective
methods (30) for extracting the active ingredients from plants
has become increasingly important. Traditionally, herbal
extracts are prepared by boiling the roots, leaves, or the entire
plant in water using a method known as tisane or decoction.
These types of extracts can be consumed orally, but these
methods are time-consuming and may not be the most effec-
tive method for extracting the active ingredients from the
herbs. This article provides a review of the state-of-the-art on
herbal processing techniques and discusses the effectiveness
of common extraction processes.
A History of the Uses of Herbs
Herbs have been used for many purposes, including as ingre-
dients for medicinal products, beauty enhancers, fragrances
and food. In the case of beauty enhancers, the natural com-
ponents in herb products are easily absorbed by the human
skin (19) and are both safe and compatible with human body,
as natural ingredients that have demonstrated no known tox-
icity. Natural products in skincare formulations have been
shown to exhibit significant antioxidant (100), emollient
(101), and UV–B protection (102) properties.
In the early nineteenth century, the first chemical analy-
ses performed by scientists were utilized to determine the
active ingredients from plant extracts, which subsequently
led to the development of natural (traditional) medicines that
had been passed historically through generations by word
of mouth (2). Many of the herbs and spices used for food
seasoning also yield useful medicinal compounds (31, 32).
Simple preparation methods for processing the plant herbs
by boiling either the entire plant or selected parts of the plant
have been employed by herbal medical practitioners for over
5,000 years (33).
STEPS IN HERBAL PROCESSING
Starting from the planting and harvesting stages, the steps
practiced for processing the natural products are very impor-
tant. Every part of the plant matures at different times,
which suggests that the harvesting process should be opti-
mized on the basis of the desired plant parts. As shown in
Table 1, the general guidelines for harvesting the three pri-
mary parts of the herbal plants (i.e., leaves, stems, and roots)
are presented to enable selection of the harvesting time on
the basis of the part of the plant that provides the opti-
mal results in the extraction process. Karimi and Jaafar (34)
showed that the leaves of L. pumila exhibited higher antiox-
idant activities and total saponin amounts (35) than the plant
roots and stems. Ramlan et al. (36) classified four differ-
ent stages of herbal processing to produce the final herbal
products.

TABLE 1
General guidelines for harvesting various plant parts
Plant part to be
collected Harvesting stage
Whole plant or Initial flowering
aerial part
Roots or rhizomes Full maturity. The maturity stage of the species must
be first determined
Leaves Fully developed leaves
Stems or bark At the beginning of the rainy season. The maturity
stage of the species must first be determined.
Wood Toward the end of the rainy season
Flowers At the beginning of the blooming period
Fruit Fully matured
Seeds Fully matured fruit
Before processing starts, the harvested herbal plants must
be pre-processed. At this stage, the plants must be dried
to remove moisture for preservation, preventing bacteria
activity and restricting fungal growth (37). To increase the
surface area of the dried herb plant materials for improved
contact with the solvent during the subsequent extraction
process, the dried plant must first be ground. An increased
surface area of the dried plant materials will improve the
performance of the extraction process. The particle size of
the milled plants is one of the factors that can affect the
extraction yield.
In the extraction processes, the operating conditions can
greatly affect the efficiency of a particular technique. For
solid–liquid extractions, important parameters include an
appropriate solvent system, the solvent-to-herbs ratio, the
particle sizes of ground and dried herbs materials, the tem-
perature, the duration, and the agitation rate, as discussed
by Sim et al. (37). Preliminary studies are necessary to
determine these operating conditions prior to starting the
extraction experiments. Literature studies can provide very
useful information for determining the optimal operating
conditions.
In some processes, solvents are mixed with herbal raw
materials (normally in the dry powder form) and then sub-
jected to forces such as heat, pressure or microwave power to
enable the phytochemicals to diffuse out of the herb cells into
the solvent medium. The mixture of solvent and extracted
phytochemicals can exist in a miscible form, or can produce
two liquid layers (if essential oil is produced). The solvents
are then removed from the mixture, typically via evapora-
tion in a rotary evaporator. Then, the crude extract is further
processed into natural products.
STATE-OF-THE-ART IN HERBAL EXTRACTION
METHODS
With herbal processing, the type of extraction process can
greatly affect the final natural products obtained. Suitable
HERBAL PROCESSING AND EXTRACTION 307
extraction processes are needed to extract the desired active
ingredients from herbs. Figure 1 shows 10 possible extrac-
tion methods that are detailed here. In Table 2, the advan-
tages and disadvantages of the available extraction methods
are presented. The biggest advantage is when the extraction
method can be used to extract plant compound in a short time
with the minimum solvent consumption. Short extraction
time is desirable due to the associated reduced electric-
ity consumption and the ability to minimize any potential
degradation of active components.
Supercritical Fluid Extraction (SFE)
Supercritical fluid extraction (SFE) is one of the extrac-
tion methods in herbal processing due its ability to extract
the valuable ingredients from herbs with high yield and
good quality (38). Its favorable features include the ability
to perform extractions at near-ambient temperatures, which
prevents thermal degradation of the substance of interest
(39). SFE is also being studied extensively as an alternative
to conventional extraction methods (38). Among the possible
SFE solvents, carbon dioxide (CO2 ) is the most established
and widely used solvent (40). CO2 has unique properties,
which include non-toxicity, non-flammability, lack of sol-
vent residue in the final product, and lack of reactivity with
extraction materials and equipment. These behaviors make it
commonly used in numerous extraction processes. In addi-
tion, this gas is less expensive than most common solvents,
which reduces the cost of the extraction process. One of the
most important benefits of CO2 in the SFE processes is the
reduction in the environmental and health concerns that arise
from the use of organic solvents (41, 42). In the SFE envi-
ronment, only one fluid phase exists; the properties of this
fluid resemble an intermediate phase between gas and liquid.
Carbon dioxide at the supercritical state has a critical tem-
perature and pressure of 31.1◦ C and 73.8 bar, respectively.
Under these conditions, CO2 exhibits both gas-like charac-
teristics, with high diffusion coefficients and low viscosities,
and liquid-like characteristics, with high solvating power
properties. These features make supercritical CO2 a good
fluid solvent (43). Both viscosity and diffusivity are factors
that affect the penetration of solvent into the herbal mate-
rials. The decreased viscosity and increased diffusivity of
solvent facilitate the penetration of solvent into porous solid
materials, resulting in increased mass transfer and reduced
extraction times in SFE (63). The solvation power, which
changes with temperature and pressure, is another factor
that can determine the effectiveness of the SFE method.
Modification of the solvent solvation power can be employed
to achieve high selectivity of the extraction product from
herbs. In addition, modification of the solvent solvation may
be useful in the extraction of complex samples, such as plant
materials (44, 45).
One of the advantages of SFE is that the system operates
at low temperatures, which is an ideal technique for studying
308 S. N. H. M. AZMIN ET AL.
1-Supercritical Fluid
Extraction (SFE)
10- Hot Water
Extraction
9- Accelerated Solvent
Extraction (ASE)
8- Ultra-high
Pressure Extraction
7- Steam-distillation
Herbal Processing and
Extraction Extraction Methods
FIGURE 1 The 10 extraction methods described in the text.
temperature-sensitive compounds (46–48) and may possi-
bly lead to the discovery of new natural compounds (49).
To minimize waste generation, the spent CO2 can be recycled
and reused, even for large-scale system. SFE is very useful
for removing pesticides from herbal medicines (50).
Unmodified CO2 may lead to a limited selectivity of this
apolar solvent. Thus, a polar solvent modifier can be used
to overcome this drawback. Significant losses of the desired
compounds can occur if an inappropriate solvent modifier is
applied. These losses can occur for desired compounds that
are removed with the solvent or via undesired reactions facil-
itated by the solvent itself (51); such losses represent two
of the major disadvantages of the SFE process. The most
commonly-used solvent modifiers are methanol, ethanol,
acetonitrile, acetone, water, ethyl ether and dichloromethane.
Methanol is a popular choice due to its effectiveness as a
polar modifier (up to 20% miscible with CO2 ). However,
the use of methanol as a modifier could be disadvantageous
when thermolabile compounds are involved because of the
slightly higher temperature required for methanol to reach
its supercritical state. In addition, a major obstacle is the cost
of purchasing the equipment compared with other methods.
Microwave-Assisted Extraction (MAE)
Microwave-assisted extraction (MAE) is a process that uses
a liquid solvent, such as water or alcohol, to extract the active
ingredients from herbs. In MAE, the enhanced extraction
occurs as the result of changes in the vegetable cell structure
2-Microwave-Assisted
Extraction (MAE)
3- Sonication
Extraction
4- Soxhlet
Extraction
5- Marinated
Extraction
6- Hydro-distillation
Extraction
caused by electromagnetic waves in MAE (52). On the other
hand, in conventional extractions, the mass transfer of the
phytochemicals occurs from inside (herbal particle) to the
outside (solvent), although the heat transfer occurs from the
outside (heat source) to the inside (herbal particle) (53).
Generally, microwaves are electromagnetic radiations with a
frequency from 0.3 to 300 GHz (54). The microwave energy
is delivered directly to the herbal particle through molecular
interactions with the electromagnetic field via conversions
of electromagnetic energy into thermal energy (55). Thus,
MAE typically results in a short extraction time and high
extraction yield. Besides, the combination of two transport
phenomena; heat and mass gradients working in the same
direction (53) also contribute to the high yield and short
extraction time of MAE.
MAE has been applied to extract the ingredients from the
leaves and roots of L. pumila (18). Laghari et al. (56) used
MAE to extract ingredients from the leaves and flowers of C.
angustifolia. The parameters that affect the extraction effi-
ciency of this extraction method are the power and frequency
of the microwaves, the duration of the microwave irradiation,
the moisture content and particle sizes of the plant samples,
the type and concentration of the solvent, the ratio of solid
to liquid, the extraction temperature, the extraction pressure,
and the number of extraction cycles (57).
Among these factors, solvent selection is the most impor-
tant parameter because the solvent affects the absorbance
of the microwave energy, as determined by the dissipation
factor (58, 59). The solvent must have an affinity for the
 HERBAL PROCESSING AND EXTRACTION 309
TABLE 2
Advantages and disadvantages of various extraction methods

Type of extraction Advantages Disadvantages

Supercritical fluid extraction, SFE
Microwave-assisted extraction, MAE
Sonication extraction
Soxhlet extraction
Marinated extraction
Hydro-distillation extraction
Steam-distillation extraction
Ultra-high pressure extraction
Accelerated solvent extraction
Hot water extraction
Solvent (CO2 ) is inexpensive
Solvent recycling can be achieved for all methods
Pure extraction yield can be attained
Applicable for both industrial and laboratory scales
Less time consuming than conventional methods
Can provide high returns on capital investment
Efficient tool for large-scale commercial
applications
Reduced working time and easy to handle
Increased yield, reduced solvent consumption, and
high extract quality
Very simple and inexpensive method
Temperature in the extraction system can be
maintained
No specific place is needed
Heat supply is not needed
Inexpensive method because of no organic solvent
is required
Can extract water insoluble compound
Short extraction time
Potential alternative technique to SFE for the
extraction of polar compounds
Reduce solvent consumption and extraction time
Inexpensive method because no organic solvent is
required
Extract essential oil with high oxygenated
components
Substantial saving of both energy and investment
cost
Loss of desired compounds with improper solvent
selection
Specialized equipment is required
Efficiency of microwaves is very poor for nonpolar target
compounds or solvents or for extremely viscous
solvents
Not appropriate for heat sensitive compounds
Expensive equipment and difficult to operate
The active part of ultrasound is only the vicinity of the
ultrasonic emitter
Presence of a dispersed phase contributes to the
ultrasound wave attenuation
Weak effect on the extraction of oil
Requires excessive extraction times
Uses large amounts of extractants (solvent)
No agitation that can accelerates the process
Thermal decomposition of heat sensitive compound
Prolonged time period is required to obtain the products
Prolonged time period is necessary to separate water
from product
No agitation to accelerate the process
Risk of thermal degradation
Energy consuming
No agitation needed to accelerate the process
Compound impurity issue
Only suitable for the extraction of high temperature
stable compound
Not suitable for heat sensitive compounds

target compound and an ability to absorb microwave energy
(60). Another critical factor in MAE is the ratio of the solid
herbs to the amount of solvent. The bioactive ingredients in
the herbs can effectively dissolve when large amounts of sol-
vent are used, thereby leading to improved extraction yields
(60, 61).
Gao et al. (61) and Li et al. (62), who applied a high sol-
vent ratio with MAE, reported consistent results. However,
their results differed from those of other studies by Guo et al.
(63), Li et al. (64), and Spigno and De Faveri (65), who
found that when large quantities of solvent were utilized, the
extraction yield initially increased and then decreased as the
solid-to-liquid ratio decreased. When the amount of solvent
was reduced (high-solid herbal materials), microwave energy
may have been absorbed and dispersed by the large amount
of plant materials (61), thereby increasing the solid mass, and
decreasing the surface area available for solvent to penetrate
the plant materials and solubilize the target molecules (66).
Agitation or shaking of the sample–solvent mixture may also
have become inadequate when the microwaves were applied
to the large-volume sample (67).
One of the advantages of MAE is the applicability of
this process for both large-scale (industrial) and small-scale
(laboratory) systems. This method is believed to be more
efficient in extracting active ingredients with less time com-
pared to conventional methods. Oil that is extracted using
MAE contains increased concentration of oxygenated com-
pounds, and have been demonstrated to exhibit antibacterial
and antifungal activities (68–70).
The primary disadvantage of MAE is the reduction of
efficiency in the microwave techniques for nonpolar target
compounds or solvents and with highly viscous solvents (57,
71). In addition, MAE is not appropriate for the extraction of
temperature-sensitive compounds (72).
310 S. N. H. M. AZMIN ET AL.
Sonication Extraction
In this extraction method, conditions such as the time and
the ratio of herbs to solvent share similarities with the MAE
processes, except the intensity of the ultrasonic equipment is
replaced instead with the microwave power to facilitate the
extraction processes. Chua et al. (19) applied a sonication
extraction technique to extract the dried leaves of L. pumila.
The correct choice of solvent coupled with appropriate agi-
tation and/or heat characteristics are also the major factors
in optimizing this type of extraction process. The solvent
extraction of herbal materials can be improved through the
use of ultrasonic power because of the mechanical effects
resulted from the ultrasound process. This effect results in
improved solvent penetration into the cellular materials via
improved mass transfer within the herbal cells. In addi-
tion, the mechanical effects disrupt the biological cell walls,
thereby resulting in the release of the cell contents such as
herbal oils (83).
In sonication processes, longitudinal waves are created
as a sonic wave encounters a liquid medium, which creates
regions of alternating compression and rarefaction (expan-
sion) among the molecules of the medium. In these regions
of changing pressure, cavitation of formed gas bubbles
occurs. During the rarefaction cycle, the surface area of these
bubbles increases, which results in increased gas diffusion
and an expansion of the bubble. In the compression cycle
(i.e., when insufficient ultrasonic energy is provided), these
vapor phases cannot remain in an expanded state, which
results in rapid condensation and the release of large amounts
of energy (73–75). The shock waves created with high tem-
peratures (550◦ C) and pressures (50 MPa) (73, 75) are a
consequence of the condensation process. Both high temper-
atures and pressures generate microjets directed toward solid
surfaces (i.e., the herbs or the container wall) as the bubbles
collapse onto those surfaces. These microjets are responsible
for the degreasing effect of ultrasound on metallic surfaces,
and this technique is commonly used to clean materials
(76).
Sonication extraction is inexpensive, an efficient tool
for large-scale commercial applications, including emul-
sification, homogenization, extraction, crystallization, low-
temperature pasteurization, degassing, defoaming, both acti-
vation and inactivation of enzymes, particle-size reduction,
and modification of viscosity (77, 78). This extraction tech-
nique, which is easy to use, features numerous advantages,
including reduced working time, increased yield, improved
solvent consumption, and possibly improved quality of the
extract. Ultrasound has weak effects on both yield and kinet-
ics for the extraction of oil such as wood seed (79). The
effects of ultrasound on the extraction yield and kinetics
may be linked to the nature of the plant matrix (57). The
presence of a dispersed phase contributes to the ultrasound
wave attenuation whereby the active part of ultrasound inside
the extractor is restricted to a zone located in the vicinity
of the ultrasonic emitter. The two factors mentioned must
be considered carefully in the design of ultrasound-assisted
extractors.
Soxhlet Extraction
Soxhlet extraction serves not only as a technique for extrac-
tion of phytochemicals, but also as a reference to compare the
newer extraction techniques (80). Soxhlet extraction involves
the use of a Soxhlet apparatus (as shown in Figure 2) for the
extraction of herbs. The sample is placed in a thimble holder
that is gradually filled with fresh solvent from a distillation
flask (see Figure 2). As the liquid reaches the overflow level,
a siphon aspirates the solute from the thimble holder, mov-
ing the aliquot back into the distillation flask and carrying
the extracted analytes into the bulk liquid. This process con-
tinues until the extraction process has been completed. The
system operates in a continuous mode as the solvent is recir-
culated through the sample. With this method, the extracts
are isolated by being filtered and the solvent is removed
at defined temperatures under reduced pressure in a rotary
evaporator (81). This extraction technique can maintain a
high system temperature (at solvent boiling point) via a heat
source applied directly to the distillation flask. This method
is also very simple and inexpensive (82).
Soxhlet extractions are limited by the time required for
the extraction process and by the large volumes of extractant
(solvent) required. The solvents can be expensive to remove,
and can cause environmental problems (83). Note that in
general, the compound boiling point is lower than that of the
solvent. Because the extraction process is usually performed
at the boiling point of the solvent for extended periods of
time, thermal decomposition of the target compounds may
also occur. In Soxhlet extraction, it is also necessary to
choose the most suitable solvent for extracting the targeted
compounds because different solvent polarity will dissolve
different compounds (84).
Hexane, which has a narrow boiling point (65◦ C) is the
most widely applied solvent. However, this solvent is listed
as an hazardous air pollutant by the U.S. Environmental
Protection Agency (85). Thus, solvent substitution is com-
monly performed. As examples, instead of using hexane,
d-limonene was used by Mamidipally and Liu (85), while
water was applied by Hanmoungjai et al. (86) to extract
oil from the rice bran. The alternative solvents, however,
result in less recovery due to the lower molecular affinity
between the alternative solvents and the solute. Sometimes,
co-solvent is added to match the polarity of the targeted
compounds. This may increase the solvent cost.
Marinated Extraction
Marinated extraction is an ancient extraction method con-
ducted at room temperature. For this technique, the parame-
ters to be considered include the type of solvent, the ratio of
dried herbs to solvent, and the extraction time. This method
requires a prolonged extraction time because it does not use

FIGURE 2 Soxhlet extraction apparatus.
heat or other forces. The extraction products are commonly
obtained by filtering the extracts first. The advantage of this
method is that it requires neither special equipment nor a spe-
cial location such as a laboratory. The commonly cited disad-
vantage of this method is the long extraction time (87). As an
example, Laghari et al. (56) extracted flowers of Cassia
angustifolia, and Zhu et al. (88) extracted Portulaca oleracea
L. within 48 hours at room temperature using this method.
Hydro-distillation Extraction
Hydro-distillation extraction techniques use water as solvent.
The mechanism for hydro-distillation extraction is similar
to that of the Soxhlet extraction process. Both methods are
commonly used to isolate volatile and non-volatile polar
components from aromatic plants (89), with small variations
in the type of solvent applied and the equipment arrange-
ment. In hydro-distillation extraction, the sample and water
solvent are placed in a retort (Figure 3). Heat is applied to
HERBAL PROCESSING AND EXTRACTION 311
heat and vaporize the mixture. Water vapor causes small sacs
in the raw plant material containing essential oil to burst.
Extracted oil is then transported by steam in the vapor
phase into the condenser where the condensate liquid mix-
ture formed. The liquid mixture then flows into a separator
where water and the essential oil are separated by density
difference. The water-rich phase, which contains some plant
essence is called “hydrosol.” As an example, when roses
are extracted using this method, the hydrosol is retained
for use as a mild antiseptic and as fragrance for floral aro-
matherapy. The experimental hydro-distillation extraction
equipment commonly used for herbal extraction is shown
in Figure 3. As a precaution, the temperature limit inside
the retort of a hydro-distillation extractor must be strictly
observed in order to prevent thermal compound degradation.
As an example, lavender essential oil must be extracted at
temperatures between 373 to 391 K (100–118◦ C). Operation
at temperatures higher than 391 K will result in the extract
containing more chemicals than the targeted aroma, causing
the therapeutic effect to be reduced. In addition, hydro-
distillation extraction must be performed within a specified
duration in order to ensure that the oil components are
completely released.
One advantage of the hydro-distillation extraction method
is that it does not require expensive organic solvent.
Nonetheless, the method involves a separation step to sep-
arate the liquid extract from water. This processing step may
require a long period of time or the use of an additional
solvent to facilitate separation (90).
Steam-distillation Extraction
Figure 4 shows the steam-distillation extraction setup.
Steam-distillation includes a steam generation apparatus to
supply steam to the mixture of solvent and plant raw mate-
rial. Steam maybe supplied at a pressure and the correspond-
ing saturation temperature that is sufficiently higher than the
boiling point of the mixture in order to allow evaporation to
take place at lower temperatures. Next, the evaporated mix-
ture of water and compound flows into a condenser where
it is condensed into a liquid mixture and later collected in
a separator. The solvent is finally evaporated and separated
from the compound using a rotary evaporator. Note that a
preliminary study is typically performed to select a suit-
able solvent to extract the phytochemicals of interest (91).
A solvent with a polarity value that is close to that of the
specific compound of interest in a herb, has a better chance
of attracting the compound.
The steam distillation-extraction method is commonly
applied for the extraction of essential oils. As steam passes
through the plant material, tiny pockets in the plant cell hold-
ing the essential oil will open and release the essential oil
without damaging the oil structure. Thus, steam-distillation
is a special extraction technique that can prevent the decom-
position of organic compound due to high temperature as its
312 S. N. H. M. AZMIN ET AL.

FIGURE 3 Hydro-distillation extraction equipment.

FIGURE 4 Steam distillation extraction setup.

temperature could be controlled. So that it could be operated
below the decomposition temperature of the compounds.
Ultra-high Pressure Extraction
High pressure in this type of extraction method refers to
cold, isostatic ultra-high hydraulic pressure. The ultra-high
pressure extraction technique involves three steps: i)
mixing herb with solvent, ii) extraction of the desired
phytochemicals at high pressure, and iii) filtering of super-
natant to remove solid particles (herbal residual).
Jun et al. (92) showed the experimental set up of this
method. Raw herbs and solvent were placed together in a
polyethylene bag and sealed after removing bubbles. Then,

the bag was placed in a pressure vessel that was equipped
with a temperature controller (at the top and the bottom of the
vessel) and pressure valve (to release the excess pressure).
Fluid was used to apply pressure to the vessel using an ultra-
high pressure booster pump. Extraction was carried out at
high pressure (100 MPa to 1,000 MPa) and at room tempera-
ture for a specified duration (5 to 15 minutes). As in the case
of other methods, after the extraction process, the mixture
was filtered and concentrated by using a rotary evaporator.
Under high pressure, the solubility of natural compounds
in any solvent is enhanced (93). This leads to the faster dif-
fusion time of compounds into the solvent medium. The
pressure range used for the UHPE method, between 100 MPa
to 1000 MPa, is the highest among the extraction methods.
For example, the pressure used in supercritical fluid extrac-
tion is about 100 MPa, while in other extraction methods
used atmospheric pressure. The ultra-high pressure applied
in UHPE leads to the increase in the compound solubility
and shorter extraction time. Xi et al. (94) compared the times
needed for the extraction of green tea leaves using differ-
ent extraction methods. In their investigation, high pressure
extraction method gave the shortest time (1 min) to extract
30% of polyphenols from green tea leaves in water.
Other methods that involved extraction at room tempera-
ture, ultrasonic extraction and heat reflux extraction needed
20 hours, 90 minutes and 45 minutes, respectively. The
results prove that the high pressure extraction method can
extract the needed compound within the shortest extraction
time. Note that, even though the UHPE method is operated
at high pressure, it requires no electrical power supply dur-
ing pressure holding. Power supply is only required when
pressure has to be increased.
Organic compounds in herbs are typically heat-sensitive
(95). Application of heat may cause the organic compounds
to denature, lose biological activity or decompose to other
compounds. Compound decomposition can be avoided using
UHPE that operates at room temperature (95). The extract
impurity is, however, a main issue and drawback of the
UHPE method. As high pressure is applied to the extraction
system, extract purity is slightly changed.
Accelerated Solvent Extraction (ASE)
Accelerated solvent extraction (ASE) is performed at ele-
vated temperatures (50 to 200◦ C) and at pressures between
10 and 15 MPa maintaining the solvent in liquid form. These
conditions tend to enhance the solvent diffusivity and accel-
erate the extraction process. Giergielewicz-Możajska et al.
(96) shows the ASE equipment used in experimental work.
There is a pressurization system that creates and applies pres-
sure to the pressure vessel. The temperature and pressure
in this system are set to be constant. The sample, together
with the solvent are placed in the closed container inside the
pressure vessel.
HERBAL PROCESSING AND EXTRACTION 313
The container is connected to a thermocouple to detect
temperature change in the sample container. If temperature
change occurs, the heating and cooling jacket (surround-
ing the sample container) will operate in order to keep
constant the temperature inside the sample container. The
pressure, on the other hand, is controlled by the pressure
relief valve. When the pressure increases, the pressure relief
valve will be opened to prevent pressure from building up.
When the pressure decreases, the pressurization system will
apply pressure to the vessel. Note that, pressure is only cre-
ated when the pressure inside the vessel drops below the set
pressure, thereby saving the pressurization energy during the
extraction process.
Note that the ASE method is normally applied for
thermally-stable organic pollutants from environmental
matrices. The ASE is not suitable for thermally labile com-
pounds because the high temperature needed in the ASE
method may lead to degradation of heat-sensitive com-
pounds. This is perhaps the reason why very few applications
of ASE have been published in the field of nutraceuticals.
The performance of ASE was proven effective by
Kaufmann and Christen (97) who compared the ASE method
with the Soxhlet extraction for isolating steroids from the
leaves of Lochroma gesnerioides. In addition, the findings
showed that both methods could produce the same results in
terms of recovery, repeatability and selectivity. The extrac-
tion time and solvent consumption may be reduced by using
the ASE (57, 97).
Hot Water Extraction
Hot water extraction (HWE) belongs to the same category
as the ASE process. The HWE however uses hot water for
extraction instead of an organic solvent. The use of water
as a solvent results in lower operating cost for the in HWE
method because water is a cheaper solvent. As a solvent,
water is also relatively easier to treat and recover, and poses
relatively less environmental issue. Ayala and De Castro (98)
proved that, the isolation of oregano essential oil using HWE
is quicker, cheaper and more effective than using hydro-
distillation extraction method. Besides, Ozel et al. (99) found
that the HWE method can extract the most valuable antioxi-
dants (carvacrol and thymol) from essential oils of Thymbra
spicata at high concentration (∼90%).
Carvacrol and thymol are valuable nutrients in the food
industry. Eikani et al. (100) also compared the efficiency of
HWE, hydro-distillation and Soxhlet extraction in extract-
ing essential oils from coriander seeds (Coriandrum sativum
L.). Their findings showed that hydro-distillation and Soxhlet
extraction showed higher extraction efficiencies than the
HWE. However, the HWE showed better results in the
extraction of more concentrated and valuable oxygenated
components in essential oils (used in food industry).
The extraction of essential oil from marjoram leaves
was done by Jimenez-Carmona et al. (101). The authors
314 S. N. H. M. AZMIN ET AL.
compared the performances of HWE and hydro-distillation
extraction processes and found that the volume of essential
oil extracted using HWE was 5.1 times the volume extracted
using hydro-distillation. The authors also reported that the
extracted essential oil contained high amount of oxygenated
components. Besides, the HWE is capable of extracting the
same amount of essential oil within 15 minutes compared to
hydro-distillation extraction, which took 3 hours as reported
by Jimenez-Carmona et al. (101) and Saim et al. (102).
Findings by the authors suggest that the HWE method is
better for extracting essential oils.
COMPARISON OF THE EXTRACTION
PROCESSES
Comparison of the extraction processes in the previous sec-
tion reveals that SFE is a well-established technique for the
extraction and separation (39) of both essential oils and its
derivatives for the food, cosmetic, pharmaceutical, and other
related industries. SFE processes also result in high-quality
essential oils with commercially viable compositions, as
compared to the products obtained using conventional steam-
distillation processes (103, 104). In the SFE method, the
physicochemical properties of the extraction fluid, such as
its density, diffusivity, dielectric constant, and viscosity, can
be easily controlled by varying the pressure or temperature
without crossing the phase boundaries (105).
Over the past four decades, supercritical CO2 has been
applied for the extraction and isolation of multiple valuable
compounds from natural products (106). The use of CO2 as a
solvent in SFE has provided an excellent alternative to chem-
ical solvents to extract the bioactive components from herbs
and other medicinal plants. The use of supercritical CO2
is also more environmentally friendly than most extraction
techniques that use organic and liquid solvents. For example,
the MAE processes use organic solvents, such as methanol
and ethanol that are relatively less environmentally friendly
than CO2 . Hawthorne et al. (107) extracted essential oils
from savory, peppermint, and dragonhead using both SFE
and steam-distillation processes.
They determined that SFE and steam-distillation extracts
had similar distributions of volatile components. The results
of their study suggested that steam-distillation did not result
in significant losses of the more volatile essential oil com-
ponents. In addition, SFE was determined to be a rapid
and quantitative method for extracting essential oils from
aromatic plants. Rezazadeh et al. (108) extracted lavender
oil and compared the efficiencies of both SFE and steam-
distillation. They determined that the extraction yield of
SFE was approximately six times higher than those obtained
using the steam-distillation process. The extraction time with
SFE was considered suitable for the processing of high qual-
ity lavender oil, compared to the products obtained using
steam-distillation processes.
Laghari et al. (56) studied the extraction of the leaves and
flowers of Cassia angustifolia using four different extrac-
tion methods including microwave, sonication, marinated,
and Soxhlet extraction processes. Among these four extrac-
tion methods, the microwave extraction method was found
to be the best technique on the basis of its ease of use,
robust processing, and minimal time required to process
the materials and to efficiently extract large amounts of
the desired phytochemicals. The extracts obtained by this
method also exhibited an increase in antioxidant activities.
The other methods investigated were effective for extracting
phytochemicals, but exhibited low efficiencies. In addition,
these other methods were found to be very difficult to handle
and needed longer periods of processing steps as compared
to the time required by the microwave extraction method.
A comparison between the sonication extraction and the
MAE indicated that the sonication extraction method was
simpler even though both methods require the same amount
of solvent. MAE, on the other hand, needed less extrac-
tion time as compared to the sonication extraction methods.
In addition, the MAE apparatus and equipment are expensive
and difficult to operate, as stated in a number of studies (57,
60, 109).
Ayala and De Castro (98), Ozel et al. (99), Eikani et al.
(100) and Jimenez-Carmona et al. (101) compared the per-
formances of hydro-distillation and HWE techniques for the
extraction of essential oils from different plants. Compared
to the hydro-distillation extraction process, they found that
the HWE offers many more advantages, including the ability
to perform separation that is:
• Quicker, (the HWE is capable of producing the same
amount of extract within 15 minutes as compared to
3 hours for the hydro-distillation extraction),
• Cheaper (the HWE uses less electricity and power
supply during the extraction process) and,
• More effective (the HWE is able to produce more
concentrated and valuable oxygenated components).
When comparing the conventional steam-distillation,
Soxhlet, and MAE methods, Chen et al. (67) suggested
that MAE had distinct advantages of decreased extraction
time, increased extraction yields, improved selectivities and
of producing superior quality targeted extract materials.
In addition, MAE was relatively cost-effective as compared
to the accelerated Soxhlet extraction process (97, 110).
Xi et al. (94) compared three extraction methods which
are sonication, marinated and ultra-high pressure to extract
polyphenols from green tea leaves. Their investigation found
that ultra-high pressure extraction method is suitable for fast
extraction of polyphenols from green tea leaves as compared
to other methods. Shouqin et al. (95) also compared the
three methods for extracting compounds from Epimedium,
green tea, propolis and Scutellaria herbs. They found that,
ultra-high pressure extraction is a very useful tool for the

extraction of natural products. The shorter extraction time
(5 to 10 minutes) is needed as the high pressure will quickly
break the plant cell and the solvent will permeate very fast
through the broken solvent. This will increase the mass trans-
fer rate of compound to the solvent area. Besides, ultra-high
pressure extraction helps save a lot of energy because the
energy is generated only when the pressure is lower than the
set pressure.
Kaufmann and Christen (97) extracted natural compound
from nine different herbs using ASE and MAE. They found
that the MAE can reduce the extraction time and solvent vol-
ume needed while ASE can increase the diffusivity of solvent
which leads to increase in the extraction speed and efficiency.
From the comparison of methods in extracting various
phytochemicals, it can be seen that researchers have been
searching for the potential methods that could result in higher
extraction yield, better selectivity, less solvent and energy
requirement, faster extraction time, as well as environmen-
tally friendly process. However, there have been works that
have compared the ten extraction methods together. The
comparison provides useful guides for future researcher and
process design engineers who may be interested to know the
relative advantages and disadvantages of the methods.
CHALLENGES FOR THE HERBAL INDUSTRY
Phytochemicals are very important constituents of herbal
products because these active components can often deter-
mine the safety and effectiveness of herbal products. Data on
phytochemical processing can be difficult to obtain, partic-
ularly the physical and chemical properties (36), including
solubility values, partition coefficients, and heat transfer
coefficients, are not available. These critical data are required
to develop an effective process model that can be integrated
into process design methods similar to those that have been
established for the petrochemical and palm-oil-based oleo-
chemical industries. The physicochemical property data can
be obtained through either experimental studies or mathe-
matical modeling efforts.
Determining the physical and chemical properties can
be challenging because herbal materials contain multiple
phytochemical components, and each component contributes
to these properties. Even with a full characterization of the
phytochemical components, the components may have iso-
mers with different properties. For example, the components
of L. pumila (Kacip Fatimah) are still uncharacterized. In the
herb extraction studies available in the literature, analysis
using gas chromatography (GC) or high-performance liquid
chromatography (HPLC) have indicated that the components
varied with the method and solvent applied during the extrac-
tion process. This lack of standardization has limited the
number of scientific studies reported for L. pumila, with
a dearth of published data on the chemical properties and
pharmacological activities of the plant extracts (81, 111).
HERBAL PROCESSING AND EXTRACTION 315
SCALE-UP METHODS
Scale-up of extraction equipment enables the commer-
cial production of herbs at larger scale to meet customer
demands. Before scaling up, the laboratory-scale method
should be tested in order to make sure that the selected
method can extract the optimum amount of phytochemicals.
According to Santos et al. (112) and Prado et al. (113), the
scale-up procedure assumed that the industrial-scale unit has
the same performance as the laboratory-scale unit when the
ratio between the mass of solid and solvent, size of raw
material plant and the operating conditions are kept con-
stant. In validating scale-up criteria, it is necessary to assess
their applicability to different types of raw materials, as the
mass transfer mechanisms may differ among species and
parts of the plant used for extraction (114). In a scale-up
plant, there is typically a huge amount of waste either from
solvent or solid waste. Thus, in scaling up the extraction
method, reusing of the waste can be very profitable (113).
As an example, grape seeds are considered a disposable
waste material by the majority of wineries. The seeds are
usually discarded, burned or used as an animal feed (115).
Santos et al. (112) compared the performances of Soxhlet
and ultra-high pressure extractions to extract phenolic com-
pounds from jabuticaba skins at laboratory scale. They found
that the ultra-high pressure extraction was more efficient than
the Soxhlet extraction method. Then, the scale up was done
for ultra-high pressure extraction. This scale-up method used
aqueous ethanol (a mixture of 70:30 v/v ethanol and water)
as solvent. Other conditions were 0.045 g/cm3 of feed to sol-
vent ratio, temperature of 353K, pressure of 5MPa and an
extraction time of only 9 minutes. Using these conditions, the
extraction resulted in 13.3% of extraction yield. The unit was
designed to operate for 8,000 hours in a year, which corre-
sponds to 330 days per year of continuous 24-hours-per-day
shift. Note that the actual operating hours of 330 instead of
365 days a year allowed for at least two days per month for
the process and plant maintenance (116).
Fiori (117) scaled up the SFE equipment for extracting
grape seed. In this plant, CO2 was used as solvent. The set
operating conditions include pressure at 50 MPa, tempera-
ture at 373.15K, ratio of solvent to feed at 4 kg CO2 /kg grape
seed and an extraction time at 2 hours. The SFE unit includes
three extraction vessels. The first vessel is for the extraction
finishing, the second is to conclude the extraction semi-cycle,
and the third is a backup vessel.
Prado et al. (113) also performed a 17-fold scale-up for
the Supercritical Fluid Extraction of grape seed. The scaled-
up conditions include a temperature of 313K, pressure of
35 MPa, ratio of CO2 solvent to feed of 8.21 kg CO2 /kg
grape seed and an extraction time of 300 minutes. The plant
was designed to operate for 24 hours with three daily shifts
for 330 days, or a total of 7920 hours of operation per year
(the rest hours was used for process and plant maintenance).
In their study, they found that higher solvent velocity tends
316 S. N. H. M. AZMIN ET AL.
to cause mechanical dragging (entrainment of oil not sol-
ubilized in CO2 ), ultimately resulting in a low yield for
the scale-up plant. In scaling-up SFE, increasing the sol-
vent mass flow will increase the entrained solvent condensed
phase. In order to prevent this phenomenon, an optimal ratio
of solvent to raw material must be maintained (118).
The 15-fold scale-up of SFE for extracting clove and sug-
arcane was successfully done by Prado et al. (119). In this
design, carbon dioxide is utilized as a solvent. All of the
conditions in the laboratory scale were set to be constant.
For the extraction of clove, the temperature is set at 313K,
pressure at 15 MPa, the ratio of solvent to feed is 3.59 kg
CO2 /kg clove and the extraction time is 130 minutes. For
sugarcane residue, the extraction conditions are at tempera-
ture of 333K, pressure of 35 MPa, ratio of solvent to feed
of 30 kg CO2 /kg sugarcane residue and an extraction time
of 360 minutes. The scale-up yield was found to be slightly
higher than that of the laboratory scale (i.e., 20% higher at
130 min for clove and 15% higher at 180 min for sugarcane
residue).
Microwave-assisted extraction was scaled up by Filly
et al. (120) to extract essential oil from rosemary. At first, the
laboratory scale extraction was performed for microwave-
assisted and hydro-distillation extractions. The result showed
that, the same extraction yield was obtained within 30 min-
utes of microwave-assisted extraction and 2 hours for hydro-
distillation extraction. Both methods actually have their
unique advantages in that, microwave gives a good yield,
while hydro-distillation gives qualitative aromatic profile.
In order to acquire both advantages, microwave-assisted
hydro-distillation was designed and scaled up.
This technique is a combination of microwave heating
and dry distillation performed at atmospheric pressure with-
out any added solvent or water. The objective of the study
is to extract the essential oil from rosemary on a labora-
tory scale and apply the same conditions to a pilot scale.
The conditions applied in the study included the use of
3 kg of dried rosemary, 30 W of microwave power and
30 minutes of extraction time without the use of any sol-
vent. The microwave-assisted hydro-distillation is a rapid
technique that consumes less energy and no solvent. It is
therefore advantageous from an environmental point of view
because no waste will be produced. The extraction schematic
is shown in Figure 5 (120).
As per 2015, there are only SFE, microwave-assisted
extraction and ultra-high pressure extraction methods in
extracting plant/ herb that have been scaled up. Another
method such as HWE was scaled up by Kilpelainen
et al. (121) to extract sawdust. The scale-up techniques
require many considerations on why the scale-up must
be done. Only promising techniques, i.e., those that
could yield a reasonably large amount of extracts within
a short time will be chosen for scale-up in order to
make economic sense and to produce a good return on
investment.
FIGURE 5 Solvent-free microwave extraction: from laboratory to pilot
scale.
CONCLUSIONS
The demand for herbal products has been increasing with the
increased awareness of possible side effects of using prod-
ucts made from chemicals. The effectiveness of natural prod-
ucts has been attributed to the presence of phytochemicals
as the active compounds in herbs. Extraction processes are
required to obtain the active ingredients that will be used in
the formulation of natural products. Appropriate solvents and
cost-effective extraction processes must be chosen to obtain
the active ingredients.
Herbal/plant extraction techniques have been typi-
cally designed to achieve one or more of the following
goals – shortened extraction time, decreased solvent con-
sumption, increased extraction yield, and enhanced extract
quality (57). The most suitable method must therefore be
chosen to perform a particular extraction task. The extraction
techniques described in this review are suitable for the solid-
liquid extraction of plant constituents. Among the common
methods to extract the essential oil are hydro-distillation,
steam-distillation, Soxhlet and marinated extraction meth-
ods. These methods are effective, but can be time-consuming
and require high operating costs. For example, Soxhlet
extraction, which uses a large amount of solvent not only
requires a high operating cost but also causes additional envi-
ronmental problems. Some other methods described in the
review required longer time to extract the needed compound
from the herbs.
For instance, the traditional method of boiling herbs
to obtain phytochemicals requires longer extraction time.
Thus, more advanced extraction methods such as sonication-
assisted extraction (122), microwave assisted extraction (97),
SFE (123), ultra-high pressure extraction (94, 95), hot water
extraction (98, 99) and ASE (97, 124) have been used as
alternatives for to perform faster extraction and obtain higher

quality yield. These techniques operate at higher tempera-
tures and/or pressures and greatly decrease the extraction
time. However, these specialized extraction techniques have
found application in limited occasions as they require higher
capital investment for the complex equipment. ASE, for
example, has been performed at high temperatures for the
extraction of organic compounds that are more stable at
high temperature relative to other compounds that tend to
denature. Research is in progress to better understand the
various extraction mechanisms with the aim to improve
their efficiencies and cost-effectiveness, to remove their lim-
itations and to perform scale up of these techniques for
commercial-scale applications.
ORCID
Lee Suan Chua http://orcid.org/0000-0003-2493-9963
FUNDING
This work was supported by the Ministry of Education,
Fundamental Research Grant Scheme (Vote number:
R.J130000.7809.4F488), Universiti Teknologi Malaysia and
the Ministry of Education, Malaysia MyPhD scholarship.
This support is gratefully acknowledged.
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