Journal of Ethnopharmacology 150 (2013) 805–817

Contents lists available at ScienceDirect

Journal of Ethnopharmacology
journal homepage: www.elsevier.com/locate/jep

Review

A review on plant-based rutin extraction methods

and its pharmacological activities
Lee Suan Chua n
Metabolites Profiling Laboratory, Institute of Bioproduct Development, Universiti Teknologi Malaysia, 81310 UTM Skudai, Johor Bahru, Johor, Malaysia

art ic l e i nf o a b s t r a c t

Article history: Ethnopharmacological relevance: Rutin is a common dietary flavonoid that is widely consumed from
Received 16 June 2013 plant-derived beverages and foods as traditional and folkloric medicine worldwide. Rutin is believed to
Received in revised form exhibit significant pharmacological activities, including anti-oxidation, anti-inflammation, anti-diabetic,
16 October 2013
anti-adipogenic, neuroprotective and hormone therapy. Till date, over 130 registered therapeutic
Accepted 17 October 2013
Available online 30 October 2013
medicinal preparations are containing rutin in their formulations. This article aims to critically review
the extraction methods for plant-based rutin and its pharmacological activities. This review provides
Keywords: comprehensive data on the performance of rutin extraction methods and the extent of its pharmaco-
Rutin logical activities using various in vitro and in vivo experimental models.
Extraction Materials and methods: Literatures including journals, patents, books and leaflets reporting on rutin from
Solid phase extraction
natural resources are systematically reviewed, particularly in the aspect of its extraction methods and
Antioxidant
biological activities. Factors affecting the efficiency of rutin extraction such as extraction temperature,
Anti-inflammation
duration and solvent to sample ratio are presented based on the findings of previous studies. The
observed biological activities followed by clear explanation are also provided accordingly.
Results: The biological activities of rutin varied largely dependent on the geographical and plant origins.
The complexity of natural rutin has impeded the development of rutin derived drugs. The detail
mechanism of rutin in human body after consumption is still unclear. Therefore, studies are intensively
carried out both in vitro and in vivo for the better understanding of the underlying mechanism. The
studies are not limited to the pharmacological properties, but also on the extraction methods of rutin.
Many studies have focused on the optimization of extraction method to increase the extraction yield of
rutin. Currently, the performances of modern extraction approaches have also been compared to the
conventional heat reflux method as a benchmark.
Conclusion: There are various extraction methods for plant-based rutin ranging from conventional
method up to the use of modern techniques such as ultrasound, mechanochemical, microwave, infrared
and pressurized assisted methods. However, proper comparison between the methods is very difficult
because of the variance in plant origin and extraction conditions. It is important to optimize the
extraction method in order to produce high yield and acceptable purity of rutin with a reasonable cost.
Even though rutin has been proven to be effective in numerous pharmacological activities, the dosage
and toxicity of rutin for such activities are still unknown. Future research should relate the dosage and
toxicity of rutin for the ethnobotanical claims based on the underlying mechanisms.
& 2013 Elsevier Ireland Ltd. All rights reserved.

Contents

1. Introduction to plant-based rutin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 806

2. Extraction methods for rutin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 806

Abbreviations: 3,4-DHPAA, 3,4-dihydroxyphenylacetic acid; 3,4-DHT, 3,4-dihydroxytoluene; 3-HPAA, 3-hydroxyphenylacetic acid; HVA, homovanillic acid (4-hydroxy-3-
ε
methoxyphenylacetic acid); AGE, advanced glycation end product; BHT, butylated hydroxytoluene; CML, N -carboxymethylysine; COX-1, cyclo-oxygenase 1; COX-2, cyclo-
oxygenase 2; DMPD, N,N-dimethyl-p-phenylendiamine; DNA, deoxyribonucleic acid; DPPH, 2,2-diphenyl-1-picrylhydrazyl; FRAP, ferric-reducing antioxidant power; ICR,
Imprinting Control Region; PRAP, phosphomolibdenum-reducing antioxidant power; RNS, reactive nitrogen species; ROS, reactive oxygen species; SPE, solid phase
extraction; TNF-α, tumor necrosis factor-α; topo I and II, topoisomerases I and II; UV, ultraviolet; WHO, World Health Organization
n
Tel.: þ 60 19 7214378; fax: þ 60 7 5569706.
E-mail addresses: lschua@ibd.utm.my, chualeesuan@utm.my

0378-8741/$ - see front matter & 2013 Elsevier Ireland Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.jep.2013.10.036
806 L.S. Chua / Journal of Ethnopharmacology 150 (2013) 805–817

2.1. Heat reflux extraction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 807

2.2. Ultrasound assisted extraction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 807
2.3. Mechanochemical assisted extraction. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 808
2.4. Microwave assisted extraction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 808
2.5. Infrared assisted extraction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 810
2.6. Pressurized liquid extraction. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 810
2.7. Solid phase extraction for rutin purification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 810
3. Pharmacological activities of rutin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 811
3.1. Antioxidant activity of rutin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 811
3.2. Anti-inflammation of rutin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 812
3.3. Medical property of rutin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 813
3.4. Hormone therapy of rutin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 814
4. Metabolism of rutin in body . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 814
5. Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 815
Acknowledgment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 815
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 815

1. Introduction to plant-based rutin

In recent decades, many herbs and natural compounds have
increasingly been receiving public interest as complementary and
alternative medicines (Ahmed et al., 2005). World Health Organi-
zation (WHO) has urged the evaluation on the effectiveness of
plant-based drugs due to the lack of scientific information
(Ayyanar et al., 2008). The natural rutin (3′,4′,5,7-tetrahydroxy-
flavone-3-rutinoside) is one of the attractive phytochemicals
because of its pharmacological activities. Therefore, it is consid-
ered as an important flavonoid in pharmaceutical industry
(Buszewski et al., 1993). Over 130 therapeutic medicinal prepara-
tions that have been registered as drugs worldwide are containing
rutin in their formulations (Reynolds, 1996; Sun and Sheng, 1998;
Erlund et al., 2000). Since the demand for natural rutin is in the
increasing trend, it is crucial to review the extraction methods and
the pharmacological activities of rutin critically.
Flavonoids are reported as the major dietary constituents of
plant-based food (Carnat et al., 1998; Goncalves et al., 2012). They
are polyphenolic compounds that occur ubiquitously in food of
plant origin because they are usually present in substantial
amount in plant kingdom. Till date, over 4000 chemically vary
flavonoids have been reported and they can be categorized into
the class of flavonols, flavones, flavanones, catechins, anthocyani-
dins and chalcones (Hollman and Batan, 1997). It is estimated
approximately 3–80 mg of flavonoids is consumed daily and
quercetin represents 50–75% of the total intake (Hertog et al.,
1995; Rimm et al., 1996). Quercetin is the aglycone moiety of rutin
after hydrolysis by the microflora in the intestines. Rutin belongs
to a kind of flavone glycoside which is also known as vitamin P.
The hydrolysis of rutin produces quercetin and rutinose catalyzed
by glucosidase (Shen et al., 2002). Quercetin usually coexists with
rutin (Chen et al., 2000) and they are mostly found in edible plants
such as onions, apples, berries, tea and wine (Manach et al., 1997).
Both rutin and quercetin are excellent sources of pharmaceutical
products for phytotherapy (Yang and Zhang, 2008).
Buckwheat (Fagopyrum esculentum Moench) from the family
Polygonaceae is reported as a major source of natural rutin (Kim
et al., 2005; Gupta et al., 2011). Significantly, the aerial part of the
plant (flowers and leaves) was found to be in the highest
concentration, approximately 2–10% of dry weight (Kalinova and
Dadakova, 2004; Suzuki et al., 2005). The similar study also
reported that young leaves contained more than 15% of rutin.
The plants contain the highest concentration of rutin in the period
of beginning flowering (Choi et al., 1996). Nevertheless, rutin could
be detected from the whole plant of buckwheat including leaves,
flowers, stalks and seeds. The amount of rutin at different parts of
the plant is greatly depended on the variance in plant species and
of its geographical origin. Indeed, the interest in rutin from
buckwheat can date back to 1940s, when buckwheat was culti-
vated as a source of rutin for medicinal use in the United States
(Ohsawa and Tsutsumi, 1995). To date, it is reported that more
than 70 plant species contain rutin. The other major commercial
sources of rutin include Ruta graveolens L. (Rutaceae), Sophora
japonica L. (Fabaceae), Maranta leuconeura E. Morren (Maranta-
ceae), Orchidantha maxillarioides (Ridl.) Schum (Lowiaceae), Stre-
litzia reginae Banks ex Aiton (Strelitziaceae), Eucalyptus spp.
(Myrtaceae), Canna indica L. (Cannaceae), Canna edulis Ker Gawl.
(Cannaceae) and Labisia pumila (Blume) Mez (Primulaceae)
(Williams and Harborne, 1977; Afshar and Delazar, 1994; Evans,
1996; Middleton et al., 2000; Abdullah et al., 2008; Chua et al.,
2011; Li et al., 2012). There was also up to 1.5% of rutin could be
extracted from tobacco leaves (Fathiazad et al., 2006).
It was found that more than half of the amount of rutin was
distributed in the upper epidermis of tartary buckwheat leaf
(Suzuki et al., 2005). The concentration of rutin also increased
rapidly after UV-B radiation. This finding has reinforced the idea
that rutin plays a role in UV screening which in good agreement
with many studies reporting flavonoids are good in protecting
plants from UV-B radiation (Harborne and Williams, 2000). On the
other hand, peroxidases such as rutin glucosidase which mostly
located in the lower epidermis will be activated under stress
condition. The activation of peroxidases is to protect plants against
oxidation by increasing the rate of respiration. Quercetin which is
produced from the hydrolysis of rutin is used as a substrate for
guaiacol peroxidases, whereas rutinose is acquired as carbohy-
drate source for respiration (Amako et al., 1994). The oxidation of
quercetin produces 3,4-dihydroxybenzoic acid which is an anti-
fungal agent for plants (Takahama and Hirota, 2000). This phe-
nomenon explains rutin and quercetin are always coexisting.
2. Extraction methods for rutin
Numerous extraction methods have been investigating in order to
extract rutin optimally from various plant samples. The effort is
driven by the renewed interest in plant-based rutin. The techniques
range from the traditional solvent extraction to modern methods
such as supercritical fluid extraction (Dimitrieska-Stojkovic and
Zdravkovski, 2003), pressurized liquid extraction (Zhang et al.,
2008; Macikova et al., 2012), microwave-assisted extraction (Zhang
et al., 2009), solid phase micro-extraction (Michalkiewicz et al., 2008)
and ultrasound-assisted extraction (Yang and Zhang, 2008). These
methods consist of two phases; liquid (solvent) and solid
 L.S. Chua / Journal of Ethnopharmacology 150 (2013) 805–817
(plant matrix) phases during extraction. Regardless the kind of
extraction method, solid–liquid extraction is a two stages of process;
(1) swelling and hydrating of plant matrix, and (2) mass transfer of
solute from plant materials to bulk solvent by diffusion and osmotic
pressure (Vinatoru, 2001). It is strongly believed that each method
has its advantages and limitations. The method of choice is highly
depended on the yield and purity of rutin.
2.1. Heat reflux extraction
Heat reflux extraction is the conventional extraction method
involving heating, boiling and refluxing for a period of time (Yang
and Zhang, 2008). This method is widely used for plant samples
extraction because the instrumentation required is simpler and
easy for operation. However, the yield of rutin might be lower due
to ionization, hydrolysis and oxidation after long duration of
extraction (Ohnishi et al., 1994; Paganga et al., 1999). The method
also requires a large volume of hazardous solvent, which is not
considered as an environmental friendly approach. Several studies
reported that a larger solvent volume could dissolve phytochem-
icals more effectively than extraction process with lower solvent to
sample ratio (Pinelo et al., 2005; Altiok et al., 2008). In a large
solvent to sample ratio, the solvent can diffuse into the plant
matrix easily and the phytochemicals can also diffuse out of the
matrix into surrounding solvent with lesser mass transfer limita-
tion. This is because extraction is a dissolution- and diffusion-
based process.
The choice of solvent is the prominent factor determining the
success of a particular extraction process, particularly for liquid–
solid extraction. According to the principle of “like dissolves like”,
solvent with the polarity value near to the polarity of target
compound is likely to be dissolved better and vice versa. This
principle explains polar rutin extraction is usually performed by
using polar alcoholic solvent such as ethanol and methanol
(Fathiazad et al., 2006). Ethanol is more preferable solvent because
it is non-toxic and cost effective solvent. Back to the year of 1924,
rutin had been extracted from the flower of elder (Sambucus
Canadensis L., Adoxaceae) using 95% alcohol by researchers from
the United States (Sando and Lloyd, 1924). In 1948, Koones and
Clifton had patented the extraction method for rutin from plant
materials using alcoholic solvent. Even though organic solvent is
the choice of solvent for rutin extraction, a small portion of water
would enhance the efficiency of extraction (Altiok et al., 2008).
Water could increase the diffusion of extractable polyphenols
through plant tissues. The swelling effect of water on plant tissues
would increase the surface area of contact between solute and
solvent (Li et al., 2004b). Kreft et al. (1999) and Kim et al. (2005)
reported that 50–60% of ethanol produced the highest yield of
rutin from buckwheat. Somehow, the variation in extraction yield
was not only due to the solubility of rutin in the solvent system,
but also the effect of the solvent system on the interaction
between rutin and buckwheat starch.
Extraction temperature and time are the other factors need to
be taken into consideration during heat reflux extraction. The high
temperature of extraction normally increases mass transfer pro-
cess. Too high extraction temperature might degrade rutin, espe-
cially under aqueous extraction system. The aqueous system
can go up to 100 1C, but mostly flavanoids are heat sensitive
compounds. Theoretically, longer extraction time produces higher
extraction yield. However, the concentration of rutin will propor-
tionally be decreasing with the length of extraction duration,
particularly when the extraction is performed at high temperature.
A single step of extraction is also not as effective as multiple
steps of extraction. The first few cycles of extraction are usually
higher in extraction yield compared to the latter extraction cycle.
Xie et al. (2011) showed that the first two cycles of extraction had
807
significantly increased the extraction yield which covered for more
than 90% of the total yield. Therefore, many researchers performed
successive extraction to increase the efficiency of extraction
for high complexity of plant samples (Altiok et al., 2008). Succes-
sive extraction could reduce the problem of diffusion for solute
migrating into bulk solvent.
2.2. Ultrasound assisted extraction
Ultrasound assisted extraction is likely to be more effective
than heat reflux method for rutin extraction. The extraction yield
of ultrasound assisted method was approximately increased for
50% at 40 1C for an hour, compared to the similar extraction
process performed by heat reflux method at 80 1C for 2 h (Zhao
et al., 1991). Interestingly, ultrasonic extraction of rutin was found
to be less suitable in aqueous medium. The apparent stability of
rutin against oxidation in methanol was relatively high compared
to water (Paniwnyk et al., 2001). The observation could be
attributed to the phenomenon of acoustic cavitation in the solu-
tion (Paniwnyk et al., 2001; Yang and Zhang, 2008). This acoustic
cavitation produced by the passage of ultrasonic wave was
significant in aqueous medium. The cavities or micro-bubbles are
produced when the ultrasound intensity is sufficient during the
expansion cycle. Free radicals that are generated within the
cavitation bubbles can produce undesirable chemical effects to
rutin extraction. Sonication in water is believed promoting the
formation of highly reactive hydroxyl radicals. The combination of
hydroxyl radicals produces hydrogen peroxide which could
degrade rutin during extraction. This phenomenon can be illu-
strated by Fig. 1. Paniwnyk et al. (2001) and Yang and Zhang
(2008) reported that the reduction in extraction yield of rutin from
Sophora japonica L. (Fabaceae) was mainly due to the degradation
rutin by the interaction with highly reactive hydroxyl radicals
during sonication in aqueous.
In contradiction to aqueous sonication, ultrasound assisted
extraction in methanol appears to be more effective in increasing
the extraction yield and reducing the extraction time (Paniwnyk
et al., 2001). Sonication in methanol does not produce a large
proportion of radicals under cavitation (Paniwnyk et al., 2001). The
cavitational effect appears to be lower because lower extraction
temperature is applied in methanol (boiling point, 64.7 1C). Further-
more, rutin is more soluble in methanol. The increase of ultrasonic
extraction efficiency in methanol could be explained by the creation
of ultrasonic wave (Paniwnyk et al., 2001). The creation of ultrasonic
wave promotes greater contact area between plant matrix and bulk
solvent. The ultrasonic wave is produced from the collapse of
cavitation bubbles that filling with solvent vapor. Before explosion,
these bubbles absorb energy from the sound waves, leading to
compression between the bubbles and the gas. The compression
increases the temperature and pressure in the solution. This
phenomenon promotes mass transfer, in addition to plant cell
Fig. 1. Acoustic cavitation produced by ultrasonic wave during extraction.
808 L.S. Chua / Journal of Ethnopharmacology 150 (2013) 805–817
disruption for the release of phytochemicals into the bulk medium
(Toma et al., 2001). Besides cell wall disruption, the reduction in
particle size and the formation of finestras in the stalks of Euonymus
alatus (Thunb.) Siebold (Celastraceae) were also observed by Yang
and Zhang (2008) from the result of scanning electron microscopy.
The induction of ultrasound caused a subsequent change in the
surface tension of cellulose, making the plant to crumble and
rupture more readily.
Besides rutin, sonication has widely been employed to extract
other compounds such as hydrocarbon (Jacques et al., 2007), fatty
acid esters (Stavarache et al., 2007), antioxidants (Albu et al., 2004),
steroids (Schinor et al., 2004) and anthraquinones (Hemwimol et al.,
2006) from plant materials. The wide application of ultrasound
assisted technique for phytochemicals extraction indirectly reveals
the feasibility and reliability of this technique for rutin separation.
Again, previous results showed that successive extraction (a three
step extraction with fresh solvent for each extraction) could produce
higher extraction yield of rutin and quercetin from Euonymus alatus
(Thunb.) Siebold (Celastraceae) by sonication (Yang and Zhang,
2008). The comparison was made between successive extraction
and single cycle of extraction with similar accumulative extraction
time (90 min).
2.3. Mechanochemical assisted extraction
Recently, a newly developed extraction technology called
mechanochemical assisted extraction has been used to efficiently
and rapidly extract rutin from Hibicus mutabilis L. (Malvaceae)
(Xie et al., 2011). The advancement of this technology is the
combination of chemical and mechanical force under solvent free
system at far lower extraction temperature. The mechanical force
is applied to explore plant cells for rutin diffusion, whereas the
chemicals are added to promote neutralization reaction between
rutin and the basic agents, namely sodium carbonate (Na2CO3) and
disodium tetraborate (Na2B4O7  10H2O). Since rutin is a weak acid
and its pKa is about 4.3, it is unstable in alkaline solution
(Jovanovic et al., 1994). This technology is of high efficiency for
compounds have low solubility in water such as rutin (12.5 g/
100 mL). The addition of solid basic agents will transform rutin
into salt form, while grinding with powdered sample matrix.
Disodium tetraborate is used to protect o-phenolic hydroxyl of
rutin from oxidation. Based on the chemical structure of rutin, it
has several acidic hydroxyl groups on its aromatic rings (Fig. 2).
The presence of these hydroxyl groups suggested that rutin could
be transformed into a highly polar compound under high energy
mechanical action (Xie et al., 2011). The minimal concentration of
solid basic agents that required for extraction is highly dependent
on the amount of target compound in the plant materials and its
conversion capacity under high energy mechanical action. There-
fore, the critical factor for high efficiency of mechanochemical
Quercetin
(Aglycone)
Rutinose
Fig. 2. Chemical structure of rutin (quercetin-3-O-rutinoside).
extraction is grinding time. Too long grinding time was reported to
reduce extraction yield resulted from rutin oxidation. Conglom-
eration might also be formed after long time of grinding under
intense mechanical action. The formation of conglomeration
would reduce the contact surface area for mass transfer, as well
as change the physiocochemical of rutin (Xie et al., 2011).
Indeed, the introduction of a water soluble borate solution to
dissolve rutin from plant materials had been reported by Koones
and Clifton (1948) long time ago. However, the extraction method
reported by them was purely chemical method because acids were
used to precipitate impurity from rutin containing extract solution.
During that time, no mechanical force was introduced to enhance
the efficiency of rutin extraction. The solubilization of rutin
increased when an adequate amount of borate solution had been
added to bring the pH of the solution to 7.2–7.5. Another 20–30%
of salting out agents such as sodium chloride, ammonium sulfate
or magnesium chloride could be added to precipitate impurities.
The remaining impurities could be further precipitated by adjust-
ing the pH of solution to 5.2–5.5 using hydrochloric acid, acetic
acid or phosphoric acid and subsequently removed from the
solution by filtration. By making the solution slightly acidic,
quercetin which is considered as the impurity for rutin extraction,
could be precipitated out. Further acidification of the filtrate with
dilute sulfuric acid to a pH of 1.0–4.0 would obtain pure rutin after
removal of impurities from the aqueous solution.
2.4. Microwave assisted extraction
Microwave energy is well known for its heating effect to
increase the rate of various analytical and biological processes
(Kaufmann and Christen, 2002; Deng et al., 2006). The significant
advantages of microwave assisted extraction are the reduction of
extraction time and the amount of solvent used (Li et al., 2004a).
According to Pare et al. (1994), microwave would induce a sudden
increase of temperature inside the cellular structure, which might
result in an eventual rupturing of cell walls and the rapid release
of plant constituents into surrounding medium. Even though the
mechanism of extraction using microwave irradiation is still
unclear, much attention has been given to the application of
microwave heating for compound extraction from herbal plants
(Kaufmann and Christen, 2002; Deng et al., 2006). This method
had been applied by Zhang et al. (2009) to extract rutin from the
samples of Euonymus alatus (Thunb.) Siebold (Celastraceae). They
reported microwave assisted extraction of rutin was better than
Soxhlet (360 min) and ultrasonic assisted extraction (30 min) in
term of shorter extraction time (6 min).
Furthermore, the high capability of ionic liquid in microwave
extraction had also been utilized for rutin extraction using micro-
wave assisted technology. Zeng et al. (2010) demonstrated that ionic
liquid-based microwave-assisted extraction was found to be higher
in extraction efficiency which was 4.88 mg/g in Schisandra chinensis
(Turcz.) Baill. (Schisandraceae) with a relative standard deviation,
1.33% and 171.82 mg/g in Flos Sophorae (Sophora japonica L., Faba-
ceae) with a relative standard deviation, 1.47% compared to those
results in both ionic liquid-based heating extraction and ultrasonic-
assisted extraction. A series of 1-butyl-3-methylimidazolium ionic
liquids with different anions were investigated and the study
reported that 1-butyl-3-methylimidazolium bromide ([bmim]Br)
aqueous solution was the best ionic liquid. As other extraction
methods, the liquid to solid ratio is the prominent factor for rutin
extraction using ionic liquids. The higher liquid to solid ratio, the
higher yield of rutin would be obtained because of sufficient contact
area between sample matrixes and ionic liquids. Even though higher
temperature reduced the viscosity of ionic liquid for mass transfer, 60
and 70 1C were the optimal temperature for rutin extraction from
Sophora japonica L. (Fabaceae) and Schisandra chinensis (Turcz.) Baill
Table 1
Pharmacological activities of rutin in different experimental models reported by previous investigators.

Biological activity Experimental model Rutin dosage Duration Reference

In vitro In vivo

Anti-oxidation Colorimetric method of hemoglobin glycosylation 0.5, 5 and 10 mg/mL 72 h Asgary et al. (1999)
Wistar albino rats 1 g/kg Korkmaz and Kolankaya
(2010)
Hyperammonemia male Wistar rats (150–200 g) induced 50 kg/kg 8 weeks Mahmoud (2012)
by ammonium chloride
Spectrophotometric antioxidant assay 0.05 mg/mL 20–150 min Yang et al. (2008)

Anti-inflammation Female Wistar rats (150–180 g) induced by adjuvant- 80 mg/kg 30 days Guardia et al. (2001)
carrageenan
Balb/c mice induced by mixture of Candida albicans & 5 mg/mL 3 times daily for an 17 days Han (2009)
Complete Freund's Adjuvant interval of 2 days
Female ICR mice (6 weeks old) hyperpermeability 10 mg/mouse 6h Lee et al. (2012)
induced by acetic acid and leukocyte migration induced

L.S. Chua / Journal of Ethnopharmacology 150 (2013) 805–817

by carboxymethylcellulose.
Human umbilical vein endothelial cells treated with 20 nM Lee et al. (2012)
lipopolysaccharide
Balb/c mice induced by lipopolysaccharide for nitric 6 mg/kg Shen et al. (2002)
oxide and prostagladin E2 production
Mouse macrophage cell line (RAW 264.7) 80 mM Shen et al. (2002)
Male Wistar rats (150–170g) induced by collagen for 25 mg/kg 21 days Umar et al. (2012)
arthritis

Anti-diabetes Fluorometric glucose glycation of collagen I-linked 150 mM 16 days Cervantes-Laurean et al.
fluorescent and non-fluorescent adduct formation (2006)
Male Sprague-Dawley rats (180–220 g) induced by 10, 30, 90 mg/kg 10 weeks Hao et al. (2012)
streptozotocin
Male Wistar strain rats 20% casein diet 4 weeks Nagasawa et al. (2002)
supplemented with 0.2%
G-rutin
Male albino Wistar rats (150–180 g) induced by 100 mg/kg 45 days Stanley Mainzen Prince
streptozotocin and Kamalakkannan
(2006)

Kidney protection Male abino Wistar rats (150–180 g) 100 mg/kg 45 days Kamalakkannan and
Stanley Mainzen Prince
(2006)

Anti-adipogenic Adipocyte in 3T3-L1 cells Dosage dependent manner Choi et al. (2006)
C57BL/6 mice fed with high-fat diet 25, 50 mg/kg 4 weeks Choi et al. (2006)

Neuroprotective Spectrophotometric enzymatic assays 50–200 mg/mL 15 min Gulpinar et al. (2012)
Male Sprague-Dawley rats (4 weeks old) induced by 0.75%w/w 2 weeks Koda et al. (2008)
trimethyltin
Male ICR mice (25–30 g) treated by dexamethasone 60 mg/kg 21 days Tongjaroenbuangam et al.
(2011)

Cardioprotection Albino Wistar rats (200–250 g) 5 and 10 mg/kg 2 weeks Annapurna et al. (2009)
Anti-cancer Weanling male Wistar rats (100 g) treated with 1 and 10 mg/100 g diet 2 weeks Webster et al. (1996)
hepatocarcinogens aflatoxin B1 and
N-nitrosodimethylamine
Anti-microorganism Agar diffusion 100–200 mg/mL methanolic DallAgnol et al. (2003)
extract of Hypericum
perforatum L. (Hypericaceae)

809
810 L.S. Chua / Journal of Ethnopharmacology 150 (2013) 805–817

(Schisandraceae), respectively (Zeng et al., 2010). The temperature

beyond optimal value would decompose rutin.

La Casa et al. (2000)

Ionic liquids are a new class of solvent composed of hetero-
Jung et al. (2007)

Guo et al. (2012)

cyclic organic cations, and inorganic or organic anions, which are
liquid near room temperature (Du et al., 2009). They are non-
Reference

molecular in nature providing intrinsic solvent properties such as

wider liquidus range, good solvation capacity, excellent microwave
absorption, excellent ionic conductivity, designable structures and
low vapor pressure, as well as high chemical and thermal stability
(Van Rantwijk and Sheldon, 2007; Macikova et al., 2012). Because
of low vapor pressure, ionic liquids are very difficult to evaporate
and they can even be recycled. Therefore, ionic liquids are getting
Duration

much attention from researchers as green solvent. Both the

7 days

selectivity and extraction efficiency were also found to be

24 h
2h

improved, particularly for extracting marker compounds from

complicated plant samples such as traditional Chinese medicine.
Several studies revealed that ionic liquids exhibited multiple
50, 100 and 200 mg/kg

interactions including hydrogen bonding, polarity, π–π, π–n and

ionic charge with rutin (Beyene et al., 2004; Shahrokhian et al.,
7.5 and 15 mg/kg

2009). The interaction had increased the solubility of rutin in ionic

Rutin dosage

liquid solution (Dursun and Nisli, 2004).

60 mg/kg

2.5. Infrared assisted extraction

Recently, infrared assisted extraction has been proven to be an

alternative method for the extraction of active components from
medicinal plants (Chen et al., 2010; Duan et al., 2010). This method
Wistar rats (180–200 g) induced by 50% ethanol
Guinea-pigs treated by aerosolized ovalbumin

uses infrared energy to heat solvent in contact with a sample in

Female Wistar rats (200 g) were ovariotized

order to partition analytes from the sample matrix into solvent.

High efficiency of heating is achieved by matching the wavelength
of infrared heater to the absorption characteristics of samples
(Duan et al., 2010). It might not as efficient as microwave assisted
extraction method in term of extraction yield, but infrared assisted
extraction is easier and cheaper as well as free of irradiation.

2.6. Pressurized liquid extraction

The unique property of ionic liquids has also widened its usage
to enhance extraction efficiency using pressurized liquid extraction
In vivo

approach. Pressurized liquid extraction is sometimes called as

accelerated solvent extraction. The advantage of this technique is
the use of high pressure to accelerate the rate of extraction from
sample matrix that packed together with inert material in a
column. The efficiency of the method can be improved by increas-
ing the number of extraction cycle, gradient profile of mobile phase
and solvent type. Wu et al. (2012) reported a novel ionic liquid-
based pressurized liquid extraction procedure coupled with high
performance liquid chromatography tandem chemiluminescence
detection to measure trace amounts of rutin and quercetin in
Chinese medicine plants including Sophora japonica L. (Fabaceae),
Crateagus pinnatifida Bunge (Rosaceae), Hypericum japonicum
Thunb. (Hypericaceae) and Morus alba L. (Moraceae). This is the
Experimental model

first contribution to utilize a combination of ionic liquid-based

pressurized liquid extraction with chemiluminescence detection.
The experimental results indicated this approach was likely to be a
promising prospect in extraction and determination of rutin and
In vitro

quercetin in medicinal plants.

2.7. Solid phase extraction for rutin purification

Biological activity
Table 1 (continued )

After extraction, further purification is usually required to

Gastroprotection
Phytoestrogen

obtain more concentrated rutin. The concentration of rutin can

Anti-asthma

be increased by removing many other plant components such as

sugars, proteins and metals (Yoon et al., 1997; Aehle et al., 2004).
As reported by Kim et al. (2005), after 1 h alcoholic extraction at
80 1C, a styrene-based resin column was used to elute rutin from
 L.S. Chua / Journal of Ethnopharmacology 150 (2013) 805–817
the buckwheat extract, and followed by rutin crystallization at 4 1C
for 12 h. This low cost process had obtained 92% of total rutin with
95% purity. The use of styrene-based resin column for rutin elution
is also called solid phase extraction (SPE).
SPE is a commonly employed method because adsorption is a
low cost separation technique. It can be a static or dynamic
adsorption and desorption process (Wang et al., 2008). In a static
SPE, a certain amount of adsorbent is added into the solution for
target compound adsorption in a batch system. After adsorption,
the solution is filtered and the target compound is then desorbed
from the adsorbent into solvent medium with the opposite
polarity to the adsorbent. The kinetic of adsorption and desorption
processes could be increased by vigorous shaking to reduce mass
transfer limitation.
Dynamic SPE is carried out in a packed column where solute is
separated using the principle of column chromatography at a
certain flow rate. Some researchers even reported that SPE was
more efficient for purification of natural constituent in term of
recovery percent through the process of adsorption and deso-
rption (Wang et al., 2008). Many synthetic adsorbents (Scordino
et al., 2003; Aehle et al., 2004; Silva et al., 2007) and biopolymers
such as collagen and celluloses had been applied to recover
polyphenolic compounds from plant extract. The efficiency of the
method is mainly determined by the chemical property of adsor-
bents (packing materials) and the composition of mobile phase as
eluent (Buszewski et al., 1993). The typical packing materials differ
based on the structure and coverage density of silica support with
alkylsilyl ligands. Previous results reported that rutin showed the
highest adsorption capacity in non-polar packing materials,
namely C18 phase (Buszewski et al., 1993). This long organosilyl
ligand phase has higher carbon percentage and lower polarity in
chemically bonded packing materials compared to C8 and C4.
This chromatographic separation technique is usually per-
formed in a vacuum manifold processor at a constant flow rate.
The SPE column needs to be preconditioned before use. The
preconditioning solvents are water and methanol for non-polar
packing materials, whereas 1,4-dioxane for polar packing materi-
als (NH2, CN and DIOL) (Buszewski et al., 1993). The adsorbent bed
is preconditioned to remove any monomers and porogenic agents
trapped inside the pores during the synthesis process.
The adsorption (qe, E) and desorption (D) capacity, as well as
recovery percent of a particular adsorbent (R) can be determined
from the equations below. The adsorption equilibrium data can be
fitted to the Henry, Langmuir and Freundlich isotherm equations
to describe the interaction of solute with the adsorbent (Jung et al.,
2001). The Henry, Langmuir and Freundlich isotherms are the
most often used isotherms for the adsorption of solute from a
solution in a separation process. The Langmuir model assumes
mono-molecular layer adsorption with a homogeneous distribu-
tion of adsorption energies and without mutual interaction
between adsorbed molecules. The Freundlich model can be used
to describe the adsorption behavior of mono-molecular layer as
well as that of the multi-molecular layer (Fu et al., 2005)
ðC o  C e ÞV o
qe ¼
m
ðC o  C e Þ
Eð%Þ ¼  100%
Co
C V
Rð%Þ ¼ d d  100%
Co V o
CdV d
Dð%Þ ¼  100%
ðC o  C e ÞV d
where qe is the adsorption capacity at equilibrium (mg/g resin);
E is the adsorption ratio (%), R is the recovery (%), D is the
811
desorption ratio (%), Co is the initial concentration of solute in
solution (mg/mL), Ce is the equilibrium concentration of solute in
solution (mg/mL), Cd is the concentration of the solute in deso-
rption solution (mg/mL), Vo is the volume of initial sample solution
(mL), Vd is the volume of the desorption solution (mL) and m is the
weight of dry resin (g).
The performance of SPE in separation can be evaluated based
on the distribution coefficient (Kd), selective coefficient (k) and
relative selectivity coefficient (k′). The adsorption process is the
distribution of the solutes between the adsorbents and the liquid
phase (Scordino et al., 2003). Kd reflects the migration and
separation capacity of the solute in two phases, k indicates the
difference of two compounds adsorbed by stationary phase or the
ratio of Kd values for two competitive compounds, whereas k′ is
defined as the ratio of K values of two competitive compounds by
different stationary phases (Zeng et al., 2012). Scatchard curve can
be plotted (qe/Ce versus qe) in order to further evaluate the binding
properties of packing materials (Eq. (8)).
ðC o  C e Þ
Kd ¼ ð5Þ
Ce
K d1
k¼ ð6Þ
K d2
kadsorbent1
k′ ¼ ð7Þ
kadsorbent2
qe
¼ ðqmax  qe ÞK d ð8Þ
Ce
3. Pharmacological activities of rutin
A lot of studies have reported the amazing physiological and
pharmacological properties of rutin in mammalian systems either
in vivo or in vitro. Table 1 summarizes the important information for
the recently reported pharmacological activities of rutin using
various experimental models as cited in this article. Most of the
biological activities such as anti-inflammation (Guardia et al., 2001;
Shen et al., 2002; Han, 2009; Umar et al., 2012), anti-microbial
(Dall'Agnol et al., 2003), anti-tumor (Ramanathan et al., 1993; Ren
et al., 2003) and anti-asthma (Jung et al., 2007) were mainly
attributed to the potent antioxidant property of rutin, particularly
as a free radical scavenger (Abraham et al., 2008; Yang et al., 2008).
Interestingly, the stability of rutin against oxidation was found to be
higher than its aglycone, quercetin (Suzuki et al., 2005). Because of
the antioxidative capacity of rutin, it is also widely used in
pharmaceutical, nutraceutical and cosmeceutical industries as a
stabilizer, preservative and natural colorant (Gonnet, 1999). It is
often used in combination with vitamin C since rutin is a bio-
flavonoid which is essential for the absorption of vitamin C and acts
as an anti-oxidizer (Buszewski et al., 1993). Human body cannot
produce bioflavonoids and rutin can be supplied through diet.
Hence, rutin is used not only for prolonging the shelf life of
ð1Þ products, but also enriching the nutritional value of products.
3.1. Antioxidant activity of rutin
ð2Þ
The strong antioxidative capacity of rutin has been proven by
numerous studies, particularly for excellent scavenging activity
ð3Þ
(Duthie and Dobson, 1999; Nagasawa et al., 2002; Abraham et al.,
2008). The scavenging activity was widely measured by 2,2-diphenyl-
ð4Þ 1-picrylhydrazyl (DPPH), N,N-dimethyl-p-phenylendiamine (DMPD),
superoxide and hydrogen peroxide produced from linoleic acid in
β-carotene bleaching assay. Besides, there are also metal-related
methods such as metal-chelating capacity and ferric-reducing
812 L.S. Chua / Journal of Ethnopharmacology 150 (2013) 805–817
antioxidant power (FRAP) and phosphomolibdenum-reducing antiox-
idant power (PRAP) assays for radical inhibition (Gulpinar et al., 2012).
These are calorimetric assays measuring the antioxidant capacity of
rutin resulted from the change of color density in the presence of
antioxidant spectrophotometrically.
Free radicals are produced during mitochondrial respiration and
also released by peroxisomes to catalyze several redox reactions of
various compounds in living tissues and cells. It is known that the
production of free radicals in human body is enhanced under certain
circumstances stimulated by external stimuli and improper diet. Free
radicals are highly reactive oxygen species (ROS) such as superoxide
(O2d ), hydroxyl (OHd ), hydrogen peroxide (H2O2), peroxyl (ROOd ),
peroxinitrite (d ONOOd ), and nitric oxide (NOd ) radicals which are
produced through oxidation within the mammalian body. They are
atom, molecule or compound that present unpaired electron. They are
produced as defense mechanism against infection caused by the
external stimuli, but excessive generation of free radical may damage
cells and tissues. The toxicity of superoxide radical (O2d ) and H2O2 in
living organisms is due to their conversion into dOH and reactive
radical metal complexes via either the iron-catalyzed Haber–Weiss
reaction or the superoxide driven Fenton reaction (Aruoma et al.,
2010). This complex is nephrotoxic and induces renal proximal tubular
damage which eventually leads to a high incidence of renal cell
carcinoma (Toyokuni, 1996). These free radicals can cause oxidative
stress and cellular damage, especially to sensitive mitochondrial
membrane phospholipids, proteins and DNA (Adibhatla and Hatcher,
2010; Montero et al., 2010). The hydroxyl radicals can access cell
membrane at the specific sites to react with DNA, and leading to cell
death and tissue damage. They initiate lipid peroxidation which is
deleterious to cell membrane by impairing membrane function
through membrane fluidity depletion and modifying membrane-
bound enzyme activity (Baynes, 1995). The formation of lipid per-
oxides by the action of free radicals on unsaturated fatty acids has
been implicated in the pathogenesis of various diseases (Mosquera
et al., 2007).
Since antioxidant is well known for its role in preventing various
pathologies (Middleton and Kandaswami, 1994), the protective
action of rutin against free radicals could be used for therapeutic
purposes (Mahmoud, 2012). Khan et al. (2009) reported that rutin
might attach to iron ions in human body in order to prevent the
metal ions from binding to hydrogen peroxides which would
otherwise create more highly reactive free radicals. The adminis-
tration of buckwheat hull extract which is known for high content
of natural rutin was reported to suppress the production of reactive
nitrogen species (RNS) such as NO2  and NO3  .
Yang et al. (2008) investigated the antioxidant mechanism of
rutin, including the total antioxidant activity, reducing power, free
radical and superoxide anion radical scavenging, hydroxyl radical
scavenging activity, and lipid peroxidation assay. At the concen-
tration of 0.05 mg/mL, the scavenging activity of rutin (90.4%
inhibition) was comparable to vitamin C (92.8% inhibition) and
approximately doubles the antioxidant activity of butylated hydro-
xytoluene (58.8% inhibition, BHT). However, the reducing power of
rutin was similar to BHT, but lower than vitamin C. The results
suggested rutin has a remarkable potency to donate electron to
reactive free radicals by converting them into more stable species
and quenching the free radical chain reaction (Yang et al., 2008).
The potent antioxidant activity of rutin is mainly due to the
presence of phenolic rings and free hydroxyl groups in the
chemical structure. These free hydroxyl groups could donate
hydrogen to prevent further oxidation.
3.2. Anti-inflammation of rutin
Inflammation is a physiological response of organism to inju-
ries such as trauma, infection or immune response. It occurs and
causes various diseases ranging from allergies to kidney failure,
stroke, cancer, colon carcinoma, asthma, rheumatoid arthritis and
many age related problems. Hence, the occurrence of chronic
diseases is mainly due to the metabolic disorder as a result of
inflammation. The inflammatory process is characterized by the
production of pro-inflammatory mediators such as eicosanoids,
ROS and cytokines. Cytokines are small protein molecules and
include interleukins (IL), lymphokines, chemokines and related
signaling molecules such as tumor necrosis factor (TNF)-α and
interferons. They are released by cells and affect cell–cell interac-
tion and communication. Both ROS and RNS might perpetuate
inflammation by facilitating the generation of chemotactic factors
at the local site (Umar et al., 2012). In particular, nitric oxide is an
important messenger molecule for inflammatory condition
(Sharma et al., 2011). It induces expression of matrix metallopro-
teinases and formation of new blood vessels which are stimulated
by prostaglandins.
Cyclo-oxygenases consisted of cyclo-oxygenase 1 (COX-1) and
cyclo-oxygenase 2 (COX-2) are the catalysts for prostaglandin synth-
esis by converting arachidonic acids to prostaglandins. COX-2 is the
inducible isomer of cyclo-oxygenase which is responsible for the
production of large amount of pro-inflammatory prostaglandins at
the inflammatory sites. Prostaglandin is also known as mediator for
the inflammatory response. High concentration of prostaglandin E2
was reported at the inflammation sites during the conversion of
arachidonic acid catalyzed by COX-2 (Shen et al., 2002). The conver-
sion also produced prostaglandin H2, prostacyclin and thomboxane
A2 as intermediates (Picot et al., 1994). Landolfi et al. (1984) reported
that quercetin, aglycone of rutin could block both cyclo-oxygenase
and lipo-oxygenase pathways at high concentration. Somehow, the
lipo-oxygenase pathway was the primary target of inflammation
inhibition at low concentration of quercetin.
There is a great interest to look for effective natural bioactive
compound in inhibiting COX-2. This enzyme is the target of
pharmacologic action of the non-steroidal anti-inflammatory
drugs (Guardia et al., 2001). Until now, most of the non-
steroidal anti-inflammatory drugs such as aspirin and indo-
methacin are specifically targeted for COX-1 inhibition. It is
hypothesized that selective inhibition of COX-2 isoenzyme might
reduce the side effects of synthetic drugs. Therefore, the identi-
fication of selective COX-2 inhibitor from natural resources has
become an important area of pharmaceutical research recently.
In most of the cases, the existing drugs are unable to prevent the
progression of inflammation, and thus leading to the irreversible
joint erosion and deformity (Guardia et al., 2001). Rutin might be
a potential candidate for COX-2 inhibition with unique mechan-
ism of action (Teresita et al., 2001). The treatment at 80 mM of
rutin produced obvious inhibitory effect on lipopolysaccharide-
induced nitric oxide production in in vitro primary peritoneal
macrophages. The similar observation was also reported for
in vivo lipopolysaccharide-injected mice at the dose of 6 mg rutin
per kg body weight (Shen et al., 2002). In the following year,
in vivo administration of rutin to the septic arthritis induced mice
was reported to exhibit 92% of inhibition on the nitric oxide
production at the dose of 20 mg rutin per mL without killing the
macrophages (Han, 2009). Most anti-arthritis drugs reduce
inflammation with the removal of macrophages aggravating
arthritis (Han, 2009). If assumed a kilogram equals to a liter,
then approximately three times increment of rutin dosage
administrated to mice could exhibit nearly 100% of inhibition in
the production of nitric oxide. Levy (1997) and Wong et al. (1998)
reported that the inhibition of COX-2 could attenuate the symp-
tom of inflammation and reduce the rate of cancer occurrence.
Therefore, the anti-inflammatory activity of rutin was found to be
beneficial for the treatment of rheumatoid arthritis and osteoar-
thritis (Umar et al., 2012).
 L.S. Chua / Journal of Ethnopharmacology 150 (2013) 805–817
The anti-arthritis property of rutin was evaluated based on the
articular elastase activity, which is known as a marker for joint
inflammation. Its activity is directly proportional to the accumula-
tion and activation of polymorphonuclear leukocytes that released
from stimulated granulocyctes from the site of injury. Apart from
enzymatic assay, the detection of nitric oxide, which is an important
signaling molecule, relates to the inflammatory response from
activated T-cells and macrophages (Han, 2009). The T-cell prolifera-
tion induces inflammatory cytokine secretion. The induction of
nitric oxide production in excess by inflammatory cytokines could
induce apoptosis in chondrocytes (Blanco et al., 1995). The bio-
chemical alteration due to arthritis was supported by histopatho-
logical observation of infected joints (Umar et al., 2012). Kauss et al.
(2008) also reported that rutin could inhibit the transcription of
more than 20 genes encoding for critical pro-inflammatory factors
including TNF-α, IL-1, IL-8 and migration inhibitory factor. Rutin
(50–100 mM) could protect vascular barrier integrity by inhibiting
hyperpermeability, expression of cell adhesion molecules, adhesion
and migration of leukocytes to suppress vascular inflammatory
diseases (Lee et al., 2012). They highlighted that the vascular barrier
protection of rutin was not due to the cytotoxicity in endothelial
cells and in mice. This was because the concentration of rutin used
in their study was below the lethal dose at 50% (950 mg/kg).
Guardia et al. (2001) reported the higher anti-inflammatory activity
of rutin than its aglycone, quecertin was because of the presence of
rutinose in the position 3 of rutin. The additional component of
rutinose had contributed to the improved activity in anti-
inflammation in term of its pharmacokinetic factor (Guardia et al.,
2001). The conclusion was made based on the inhibition percentage
of adema pedal after intraperitoneal administration of rutin at the
dose of 80 mg/kg to female Wistar rats from day 1 to day 30.
Interestingly, rutin was also found to be effective in inhibiting
Candida albicans and resulted in no hemolysis (Han, 2009). The
findings indicated that rutin could be used for both anti-arthritic
and anti-candidal treatment caused by the yeast cells. The result was
in line with the finding of Handa et al. (1992) who reported flavonoids
displayed significant activity in both proliferative and exudative phases
of inflammation. Mostly, flavonoids affect non-specific immunological
responses (acute inflammatory reaction) by suppressing macrophage
phagocytosis, releasing oxidant by neutrophils and activation of mast
cells. Only a small portion of flavanoids exhibit complex biphasic
action towards the specific immunological system. It is noteworthy
that flavonoids appear to be function at low concentration, but
inhibitory at high concentration (Formica and Regelson, 1995).
3.3. Medical property of rutin
Although there are many studies relating to biological activities
of rutin, the mechanism of these activities are still unclear (Shen
et al., 2002). This explains the application of rutin in treating
human diseases is still uncommon in western medication (Hao
et al., 2012). This might be due to the high effective concentration
and poor absorption of rutin after oral intake (Shen et al., 2002).
Nevertheless, rutin has been proven to be effective in reducing the
risk of chronic diseases (Knekt et al., 2002). This prevalence
accelerates the elucidation of their underlying mechanism, espe-
cially for medical application of rutin.
The anti-diabetic property of rutin had been proven by the
Stanely Mainzen Prince group on the treatment of diabetic mellitus
by improving glucose homeostasis of diabetic rats (Stanley Mainzen
Prince and Kamalakkannan, 2006). The homeostasis of glucose was
achieved by increasing insulin level, and increasing glycogen con-
tent in liver and muscle, but decreasing the glycogen content in
kidney. The fasting plasma glucose was reduced by increasing the
activity of hexakinase, but decreasing the activities of glucose-
6-phosphatase and fructose-6-bisphosphatase in the tissues.
813
Therefore, rutin was found to have anti-diabetic property (Ushida
et al., 2008; Han, 2009). Cervantes-Laurean et al. (2006) explained the
anti-diabetic property of rutin was due to its vicinyl hydroxyl group-
containing metabolites for early glycation product formation inhibi-
tion. The serum and kidney proteins of diabetic rats showed 20% of
ε
reduction in the amount of N -fructoselysine after 4 weeks of feeding
the Wistar strain rats with rutin rich diet (Nagasawa et al., 2002).
Glycation is a reversible and non-enzymatic reaction of the aldehyde
groups of reducing sugars with amino groups of proteins. The products
from glycation due to excessive glucose can chemically modify DNA
causing mutation and complex DNA rearrangement. A variety of
sugars including glucoses, glucose autoxidation products (arabinose
and glyoxal) and pentoses can also contribute to advanced glycation
end products (AGEs) formation (Cervantes-Laurean et al., 1996). In
addition to sugars, oxidation of lipids and amino acids can also result
ε
in N -carboxymethylysine (CML) formation, especially during inflam-
mation. This non-enzymatic glycosylation of protein occurs between
reducing sugars and primary amino groups in protein by direct
reaction (Asgary et al., 1999). The reaction forms Schiff bases, followed
by Amadori rearrangement to yield a stable ketoamine derivative
of protein (Lee and Cerami, 1992). The ketoamine will then form a
variety of fluorescent and non-fluorescent AGEs through oxidation
(Cervantes-Laurean et al., 2006). Pentosidine is an example of fluor-
escent AGE, whereas CML is non-fluorescent AGE (Cervantes-Laurean
et al., 2006). CML and pentosidine are AGEs that are increased in skin
collagen I during both intrinsic aging and diabetes (Dyer et al., 1993).
The extent of AGE formation is increased during diabetic hyperglyce-
mia (Asgary et al., 1999). These AGEs always associate with numerous
pathologies. Rutin metabolites were found to be able inhibiting
glucose-induced collagen fluorescent relevant to hyperglycemia
(Vishwanath et al., 1986). Both rutin and its aglycone (quercetin)
extracted from the stalks of Euonymus alatus (Thunb.) Siebold
(Celastraceae) were found to exhibit identical therapeutic potency
in treating diabetes (Yang and Zhang, 2008). Besides rutin and
quercetin, its vicinyl dihydroxyl groups containing metabolites such
as 3,4-dihydroxyphenylacetic acid (3,4-DHPAA), 3,4-dihydroxyto-
luene (3,4-DHT) had also been proven to inhibit the formation of
pentosidine and fluorescent adducts, as well as glycation of collagen I
in a dose-dependent manner. The non-vicinyl dihydroxyl group
containing metabolites, namely homovanillic acid (HVA) and m-3-
hydroxyphenylacetic acid (m-3-HPAA) were less effective in CML
formation (Cervantes-Laurean et al., 2006).
Prolong diabetic problem always accompanied by kidney
damage due to the deterioration of kidney function for excessive
glucose filtration. The same group of researchers also showed that
rutin could protect the kidney of diabetic rats by decreasing the
accumulation of hydroxyproline, laminin and type IV collagen, as
well as decreasing the tissue inhibitors of metalloproteinases, but
increasing the activity of matrix metalloproteinases in kidney
(Kamalakkannan and Stanely Mainzen Prince, 2006). By increasing
the activity of matrix metalloproteinases, the enzymes are capable
to degrade all kinds of extracellular matrix proteins. It is known that
a variety of proteins are also subjected to non-enzymatic glycation
which is consequently contributed to various long-term complica-
tion of the disease.
All these results indicated that rutin might postpone renal
damage and might be a potential drug for the prevention of early
diabetic neuropathy (Hu et al., 2009; Hao et al., 2012). It could
prevent glycosylation of hemoglobin from getting serious which
might then lead to other complication of diabetes such as nerve
damage and blindness (Asgary et al., 1999). Rutin was able to
reverse the trimethyltin induced spatial memory impairment and
the damage to pyramidal neurons in the hippocampal region. The
protective effect of rutin (0.75%w/w) had been proven by Koda et al.
(2008) from their in vivo study on 4 weeks old of male Sprague-
Dawley rats. A group of researchers from Thailand reported that
814 L.S. Chua / Journal of Ethnopharmacology 150 (2013) 805–817
the administration of rutin at 60 mg/kg body weight to the
dexamethasone-treated male ICR (Imprinting Control Region) mice
had successfully reversed cognitive deficits including impaired
dentate gyrus cell proliferation, and protected against morphologi-
cal changes in the CA3 region (Tongjaroenbuangam et al., 2011).
In vitro neuroprotective property of rutin was also reported by
Gulpinar et al. (2012) using enzymatic methods such as acetylcho-
linesterase, butyrylcholinesterase and tyrosinase inhibition assays
spectrophotometrically. These might explain the findings of other
investigators who reported the positive effect of rutin for the
improvement of sight and hearing capability (Campbell, 1997),
hypertension (Ushida et al., 2008; Lee et al., 2012), hepatotoxicity
(Janbaz et al., 2002) and memory impairment (Pu et al., 2007;
Han, 2009). The high antioxidative activity of rutin could inhibit the
glycosylation of protein, but its performance in the prevention of
protein glycosylation was somehow lower than its aglycone, quer-
cetin. The good performance of quercetin was due to the existence
of o-dihydroxyl groups or non-glycosylation of hydroxyl group in
the chemical structure.
There were also investigators who proved that rutin could reduce
blood fat and cholesterol by decreasing the levels of lipids, particu-
larly the low density and very low density lipoprotein cholesterol in
plasma, but increase the levels of plasma high density lipoprotein
cholesterol (Kayashita et al., 1997; Zeng et al., 2010). The findings
were in line with Jiang et al. (2007) who reported a dose response
effect of rutin (0.025–8 mg buckwheat/mL) in inhibiting low density
lipoprotein peroxidation. Moreover, the development of fatty liver by
body weight gain in mice treated with high fat plus rutin diet (25 and
50 mg/kg body weight daily) had been proven to be lower than those
mice (C57BL/6) treated with high fat diet only, 64.4% of the total
calories as fat for 4 weeks (Choi et al., 2006). Rutin even quoted as a
promising flavanoid to reduce the risk of atherosclerosis because of
its capacity in inhibiting low density lipoprotein oxidation
(Milde et al., 2004). Rutin was reported to be effective in reducing
abnormal leakage, capillary impairment and venous insufficiency in
cardiovascular diseases (Hertog et al., 1995; Reynolds, 1996; Rimm
et al., 1996; Annapurna et al., 2009). It showed the inhibition of
human platelet aggregation which might cause stroke, myocardial
infarction, pulmonary embolism or the blockage of blood vessels to
other parts of the body (Pace-Asciak et al., 1995). The blood vessel
hardening condition in atherosclerosis could subsequently lead to
cardiovascular diseases. Korkmaz and Kolankaya (2010) reported that
the effective treatment for fragility of blood vessel capillary was due
to the high radical scavenging activity and antioxidant capacity of
rutin. This quercetin glycoside; rutin has been recognized for its
capability to decrease the permeability of capillaries.
Another well known biological activity of rutin is its anti-cancer
property. The anti-cancer property of rutin was observed from the
inhibition of various cancer cell lines in vitro and the reduction of
tumor development in experimental animals (Deschner et al., 1991;
Webster et al., 1996; Van der Logt et al., 2003). The reduction might
be attributed to the inhibition of DNA topoisomerases I and II (topo I
and topo II), which are the markers of DNA and chromosome
damage (Cantero et al., 2006). Indeed, cancer cells are produced in
order to protect body from external stimuli. The aggressive growth
of cancer cells because of weak immune system against the external
stimuli has caused the killing effect. Other biological activity of rutin
includes gastroprotective effect against gastric lesion and gastric
mucosal ulceration (La Casa et al., 2000).
3.4. Hormone therapy of rutin
Based on the previous studies, plants which have the compound
with almost similar structure of rutin such as daidzein and
formononetin were found to be able regulating the endocrine
system of body (Guo et al., 2012). They are phytoestrogens which
act as estrogens. Estrogen is one of the main hormones and the
development of mammary glands is mainly regulated by neuro-
endocrine system (Tucker, 1981). Therefore, rutin is also a phytoes-
trogen which could bind with estrogen receptor and then exert
estrogen-like effects because of the structural similarity to endo-
genous estrogen (Suman and Saffron, 2008). Rutin has double-
glycoside structure that formed through the combination of hydro-
xyl at position C3 of quercetin and rutinose (Guo et al., 2012). This
compound has a similar structure of planar double benzene rings to
the endogenousestrogen17-β-estradiol (17-β-E2). It is considered
that this structure could combine to the site of estrogen receptor
occupied by 17-β-E2 and thus play an estrogen-like role (Tham
et al., 1998).
4. Metabolism of rutin in body
Although rutin has been reported to be widely consumed from
edible plants, its precise metabolism is still unknown. Rutin, as
other flavonoids usually occur as glycosides in dietary plants
(Manach et al., 1997). It is generally considered that flavonoid
glycosides are firstly hydrolyzed by the digestive microflora before
being absorbed (Kuhnau, 1976). Therefore, rutin was found to be
absorbed more slowly than quercetin because quercetin was ready
available for digestive absorption both in the small intestine and in
the large bowel (Manach et al., 1997). However, Hollman et al.
(1995) reported rutin from onion was more readily absorbed than
its aglycone moiety in ileostomy patients. Therefore, the charac-
terization and structure–function relationship of rutin and its
metabolites are of great importance for better understanding of
the role of rutin in disease prevention and progression (Cervantes-
Laurean et al., 2006). The understanding about the mechanism of
rutin is essential for the evaluation of possible physiological effects
of rutin (Manach et al., 1997). In recent years, the mechanism of
flavonoid glycosides are gradually understood that they are gen-
erally hydrolyzed by intestinal and bacterial enzymes to corre-
sponding aglycones and other smaller metabolites which is
absorbable by the gut (Hollman and Batan, 1997).
Studies reported that little or no dietary rutin is absorbed
because gut microflora in the intestines metabolize rutin into a
variety of small metabolites (Griffiths and Barrow, 1972; Winter
et al., 1989). The reaction involves hydrolysis of rutin catalyzed by
α-rhamnosidase and β-glucosidase (Bokkenheuser et al., 1987;
Manach et al., 1995). The metabolites produced include quercetin
(3,5,7,3′,5′-pentahydroxyflavonol), isoquercetin (quercetin 3-glyco-
side) and other phenol derivatives such as 3,4-DHPAA, 3,4-DHT,
3-HPAA, and HVA (Griffiths and Barrow, 1972; Baba et al., 1981;
Winter et al., 1989; Schneider et al., 2000; Braune et al., 2001;
Cervantes-Laurean et al., 2006; Arjumand et al., 2011). Manach et al.
(1996) reported that about 80% of circulating plasma quercetin is
present as methoxy derivative (isorhamnetin), and flavonol meta-
bolites were circulating in plasma as conjugated derivatives. Con-
jugation or methylation arises from the presence of two –OH on the
B-ring, or the presence of –OH in the position of 3 and 5 at the
vicinity of the 4-oxo group probably plays a role in lowering the
reactivity of quercetin (Manach et al., 1997). These metabolites,
particularly quercetin, 3,4-DHPAA and 3,4-DHT contain vicinyl
dihydroxyl groups in their chemical structure which are important
for oxidation inhibition. The presence of vicinyl dihydroxyl groups
was also shown to affect the phenols inhibiting iron and copper-
catalyzed production of radical species (Cervantes-Laurean et al.,
2006). It is likely that metal chelation and free radical scavenging
properties contributed to the inhibition of glucose autoxidation by
rutin metabolites containing vicinyl dihydroxyl groups.
Quercetin plasma levels have been found to be 3.5 μmol/L,
when doses of 50 mg of either rutin or quercetin were used in the
 L.S. Chua / Journal of Ethnopharmacology 150 (2013) 805–817
diet of healthy human volunteers (Erlund et al., 2000). Moreover,
up to 50% of an ingested dose of 75 mg rutin was recovered as
microbial metabolites from urine of human volunteers (Sawai
et al., 1987). This finding further supports the appearance of
micromole per liter metabolite concentration after rutin consump-
tion. Rutin is hydrolyzed into low molecular weight phenolic acids
besides its aglycone, quercetin by the colonic microflora (Olthof
et al., 2003). This is because the polyphenol rutin derivatives like
quercetin-3-o-rhamnoglucoside were not found in urine after
consumption. However, 3-HPAA, HVA, 3,4-DHPAA, and 3,4-DHT
which are the metabolites resulted from colonic microflora degra-
dation were detected (Baba et al., 1981).
5. Conclusion
Rutin appears to be a potential phytochemical ingredient in
food supplement and medicinal products nowadays. Numerous
studies have reported the diverse pharmacological activities of
rutin, as well as the risk reduction of diseases for health promo-
tion. Owing to its significant functionality, the incorporation of
rutin into food-based products is likely to be a promising practice
for development of functional foods and nutraceuticals nowadays
(Zhu et al., 2008). Somehow, the incorporation requires scientifi-
cally proven data for the ethnobotanical claims. On the one hand,
biologists, botanists and pharmacists are actively involved in the
studies in relation to biological activities of rutin both in vitro and
in vivo. The biochemical properties and metabolic changes after
rutin consumption are also being investigated intensively. On the
other hand, technologists and engineers are focused on the
optimization of rutin extraction and production. With the involve-
ment of multidisciplinary profession, the mechanism of rutin will
be clearly explained with the efficacy and safety data, whereas
high yield of rutin can be obtained from the well optimized
extraction method with cost effective operation approach in near
future.
Acknowledgment
The author would like to express her gratitude to Research
Alliance of Biotechnology from Universiti Teknologi Malaysia for
providing the research grant GUP (Tier 2) 05J84.
References
Abdullah, Y., Schneider, B., Petersen, M., 2008. Occurrence of rosmarinic acid,
chlorogenic acid and rutin in Marantaceae species. Phytochem. Lett. 1,
199–203.
Abraham, L.C.N., Masakuni, T., Isao, H., Hajime, T., 2008. Antioxidant flavonoid
glycosides from the leaves of Ficus pumila L. Food Chem. 109, 415–420.
Adibhatla, R.M., Hatcher, J.F., 2010. Lipid oxidation and peroxidation in CNS health
and disease: from molecular mechanisms to therapeutic opportunities. Anti-
oxid. Redox Signaling 12, 125–169.
Aehle, E., Grandic, S.R.L., Ralainirina, R., Baltora-Rosset, S., Mesnard, F., Prouillet, C.,
Maziere, J.C., Fliniaux, M.A., 2004. Development and evaluation of an enriched
natural antioxidant preparation obtained from aqueous spinach (Spinacia
oleracea) extracts by an adsorption procedure. Food Chem. 86, 579–585.
Afshar, J., Delazar, A., 1994. Rutin from Ruta graveolens L. J. Sch. Pharm. (Med. Sci.
Univ. Tehran) 4, 1–12.
Ahmed, S., Anuntiyo, J., Malemud, C.J., Haqqi, T.M., 2005. Biological basis for the use
of botanicals in osteoarthritis and rheumatoid arthritis: a review. Evidence-
Based Complementary Alternative Med. 2, 301–308.
Albu, S., Joyce, E., Paniwnyk, L., Lorimer, J.P., Mason, T.J., 2004. Potential for the use
of ultrasound in the extraction of antioxidants from Rosmarinus officinalis for
the food and pharmaceutical industry. Ultrason. Sonochem. 11, 261–265.
Altiok, E., Baycin, D., Bayraktar, O., Ulku, S., 2008. Isolation of polyphenols from the
extracts of olive leaves (Olea europaea L.) by adsorption on silk fibroin. Sep.
Purif. Technol. 62, 342–348.
Amako, K., Chen, G., Asada, K., 1994. Separate assays specific for ascorbate
peroxidase and guaiacol peroxidase and for the chloroplastic and cytosolic
isozymes of ascorbate peroxidase in plants. Plant Cell Physiol. 35, 497–504.
815
Annapurna, A., Reddy, C.S., Akondi, R.B., Rao, S.R.C., 2009. Cardioprotective actions
of two bioflavonoids, quercetin and rutin, in experimental myocardial infarc-
tion in both normal and streptozotocin-induced type I diabetic rats. J. Pharm.
Pharmacol. 61, 1365–1374.
Arjumand, W., Seth, A., Sultana, S., 2011. Rutin attenuates cisplatin induced renal
inflammation and apoptosis by reducing NFkappaB, TNF-alpha and caspase-3
expression in Wistar rats. Food Chem. Toxicol. 49, 2013–2021.
Aruoma, O.I., Hayashi, Y., Marotta, F., Mantello, P., Rachmilewitz, E., Montagnier, L.,
2010. Applications and bioefficacy of the functional food supplement fermen-
ted papaya preparation. Toxicology 278, 6–16.
Asgary, S., Naderi, G., Sarrafzadegan, N., Ghassemi, N., Boshtam, M., Rafie, M.,
Arefian, A., 1999. Anti-oxidant effect of flavonoids on hemoglobin glycosylation.
Pharm. Acta Helv. 73, 223–226.
Ayyanar, M., Sankarasivaraman, K., Ignacimuthu, S., 2008. Traditional herbal
medicines used for the treatment of diabetes among two major tribal groups
in South Tamil Nadu, India. Ethnobot. Leafl. 12, 276–280.
Baba, S., Furuta, T., Horie, M., Nakagawa, H., 1981. Studies on drug metabolism by
use of isotopes. XXVI. Determination of urinary metabolites of rutin in humans.
J. Pharm. Sci. 70, 780–782.
Baynes, J.W., 1995. Reactive oxygen in the aetiology and complications of diabetes.
In: Ioannides, C., Flatt, P.R. (Eds.), Drug Diet and Disease, Vol. 2., Mechanistic
Approach to Diabetes. Ellis Horwood Limited, Hertfortshive, pp. 230–231.
Beyene, N.W., Kotzian, P., Schachl, K., Alemu, H., Turkusicd, E., Coprad, A.,
Modereggera, H., Svancara, I., Vytras, K., Kalcher, K., 2004. (Bio)sensors based
on manganese dioxide-modified carbon substrates: retrospections, further
improvements and applications. Talanta 64, 1151–1159.
Blanco, F.J., Ochs, R.L., Schwarz, H., Lotz, M., 1995. Chondrocyte apoptosis induced
by nitric oxide. Am. J. Pathol. 146, 75–85.
Bokkenheuser, V.D., Shackleton, C.H., Winter, J., 1987. Hydrolysis of dietary
flavonoid glycosides by strains of intestinal bacteriodes from humans. Biochem.
J. 248, 953–956.
Braune, A., Gutschow, M., Engst, W., Blaut, M., 2001. Degradation of quercetin and
luteolin by Eubacterium ramulus. Appl. Environ. Microbiol. 67, 5558–55567.
Buszewski, B., Kawka, S., Suprynowicz, Z., Wolski, T., 1993. Simultaneous isolation
of rutin and esculin from plant material and drugs using solid-phase extraction.
J. Pharm. Biomed. Anal. 11, 211–215.
Campbell, C.G., 1997. Buckwheat. Fagopyrum esculentum Moench. Promoting the
conservation and use of underutilized and neglected crops, vol. 19. 19. Institute
of Plant Genetics and Crop Plant Research, Gatersleben/International Plant
Genetic Resources Institute, Rome, Italy.
Cantero, G., Campanella, C., Mateos, S., Cortes, F., 2006. Topoisomerase II inhibition
and high yield of endoreduplication induced by the flavonoids luteolin and
quercetin. Mutagenesis 21, 321–325.
Carnat, A.P., Carnat, A., Fraisse, D., Lamaison, J.L., Heitz, A., Wylde, R., Teulade, J.C.,
1998. Violarvensin, a new flavone di-C-glycoside from Viola arvensis. J. Nat.
Prod. 61, 272–274.
Cervantes-Laurean, D., Jacobson, E.L., Jacobson, M.K., 1996. Glycation and glycox-
idation of histones by ADP-ribose. J. Biol. Chem. 271, 10461–10469.
Cervantes-Laurean, D., Schramm, D.D., Jacobson, E.L., Halaweish, I., Bruckner, G.G.,
Boissonneault, G.A., 2006. Inhibition of advanced glycation end product
formation on collagen by rutin and its metabolites. J. Nutr. Biochem. 17,
531–540.
Chen, G., Zhang, H., Ye, J., 2000. Determination of rutin and quercetin in plants by
capillary electrophoresis with electrochemical detection. Anal. Chim. Acta 423,
69–76.
Chen, Y.L., Duan, G.L., Xie, M.F., Chen, B., Li, Y., 2010. Infrared-assisted extraction
coupled with high-performance liquid chromatography for simultaneous
determination of eight active compounds in Radix Salviae miltiorrhizae. J. Sep.
Sci. 33, 2888–2897.
Choi, B.H., Kim, S.L., Kim, S.K., 1996. Rutin and functional ingredients of buckwheat
and their variations. Korean J. Crop Sci. 41, 69–93.
Choi, T., Park, Y., Choi, H., Lee, E.H., 2006. Anti-adipogenic activity of rutin in 3T3-L1
cells and mice fed with high-fat diet. Biofactors 26, 273–281.
Chua, L.S., Latiff, N.A., Lee, S.Y., Lee, C.T., Sarmidi, M.R., Aziz, R.A., 2011. Flavonoids
and phenolic acids from Labisia pumila (Kacip Fatimah). Food Chem. 127,
1186–1192.
Dall'Agnol, R., Ferraz, A., Bernardi, A.P., Albring, D., Nor, C., Sarmento, L., Lamb, L.,
Hass, M., von Poser, G., Schapoval, E.E.S., 2003. Antimicrobial activity of some
Hypericum species. Phytomedicine 10, 511–516.
Deng, C.H., Yao, N., Wang, B., Zhang, X.M., 2006. Development of microwave-
assisted extraction followed by headspace single-drop microextraction for fast
determination of paeonol in traditional Chinese medicines. J. Chromatogr. A
1103, 15–27.
Deschner, E.E., Ruperto, J., Wong, G., Newmark, H.L., 1991. Quercetin and rutin as
inhibitors of azoxymethanol-induced colonic neoplasia. Carcinogenesis 12,
1193–1196.
Dimitrieska-Stojkovic, E., Zdravkovski, Z., 2003. Supercritical fluid extraction of
quercetin and rutin from Hyperici Herba. J. Liq. Chromatogr. Relat.Technol. 26,
2517–2533.
Du, F.Y., Xiao, X.H., Luo, X.J., Li, G.K., 2009. Application of ionic liquids in the
microwave-assisted extraction of polyphenolic compounds from medicinal
plants. Talanta 78, 1177–1184.
Duan, H.T., Chen, Y., Chen, G., 2010. Far infrared-assisted extraction followed by
capillary electrophoresis for the determination of bioactive constituents in the
leaves of Lycium barbarum Linn. J. Chromatogr. A 1217, 4511–4516.
816 L.S. Chua / Journal of Ethnopharmacology 150 (2013) 805–817
Dursun, Z., Nisli, G., 2004. Voltammetric behavior of copper(I)oxide modified
carbon paste electrode in the presence of cysteine and ascorbic acid. Talanta
63, 873–878.
Duthie, S.J., Dobson, V.L., 1999. Dietary flavonoids protect human colonocyte DNA
from oxidative attack in vitro. Eur. J. Nutr. 38, 28–34.
Dyer, D.G., Dunn, J.A., Thorpe, S.R., Bailie, K.E., Lyons, T.J., McCance, D.R., Baynes, J.
W., 1993. Accumulation of Maillard reaction products in skin collagen in
diabetes and aging. J. Clin. Invest. 91, 2463–2469.
Erlund, I., Kosonen, T., Alfthan, G., Maenpaa, J., Perttunen, K., Kenraali, J., Para-
ntainen, J., Aro, A., 2000. Pharmacokinetics of quercetin from quercetin
aglycone and rutin in healthy volunteers. Eur. J. Clin. Pharmacol. 56, 545–553.
Evans, W.C., 1996. Trease and Evans Pharmacognosy, 14th Ed. WB Saunders
Company Ltd., London p. 251.
Fathiazad, F., Delazar, A., Amiri, R., Sarker, S.D., 2006. Extraction of flavonoids and
quantification of rutin from waste tobacco leaves. Iran. J. Pharm. Res. 3,
222–227.
Formica, J.V., Regelson, W., 1995. Review of the biology of quercetin and related
bioflavonoids. Food Chem. Toxicol. 33, 1061–1080.
Fu, B., Liu, J., Li, H., Li, L., Lee, F.S.C., Wang, X., 2005. The application of macroporous
resins in the separation of licorice flavonoids and glycyrrhizic acid. J. Chroma-
togr. A 1089, 18–24.
Goncalves, A.F.K., Friedrich, R.B., Boligon, A.A., Piana, M., Beck, R.C.R., Athayde, M.L.,
2012. Anti-oxidant capacity, total phenolic contents and HPLC determination of
rutin in Viola tricolor (L) flowers. Free Radicals Antioxid. 2, 32–37.
Gonnet, J.F., 1999. Colour effects of co-pigmentation of anthocyanins revisited-2. A
colorimetric look at the solutions of cyanin copigmented by rutin using the
CIELAB scale. Food Chem. 66, 387–394.
Guardia, T., Rotelli, A.E., Juarez, A.O., Pelzer, L.E., 2001. Anti-inflammatory properties
of plant flavonoids. Effects of rutin, quercetin and hesperidin on adjuvant
arthritis in rat. Farmaco 56, 683–687.
Gulpinar, A.R., Orhan, I.E., Kan, A., Senol, F.S., Celik, S.A., Kartal, M., 2012. Estimation
of in vitro neuroprotective properties and quantification of rutin and fatty acids
in buckwheat (Fagopyrum esculentum Moench) cultivated in Turkey. Food Res.
Int. 46, 536–543.
Guo, X., Diao, Q., Wang, Y., Tu, Y., Deng, K., Wang, X., Fu, T., Yan, G., 2012. The effect
of administration of rutin on plasma levels of estrogen, prolactin, growth
hormone and gene expression of their receptors in mammary glands in
ovariectomized rats. J. Integrative Agric. 11, 1700–1706.
Gupta, N., Sharma, S.K., Rana, J.C., Chauhan, R.S., 2011. Expression of flavonoid
biosynthesis genes vis-a-vis rutin content variation in different growth stages
of Fagopyrum species. J. Plant Physiol. 168, 2117–2123.
Griffiths, L.A., Barrow, A., 1972. Metabolism of flavonoid compounds in germ-free
rats. J. Biochem. 130, 1161–1162.
Han, Y., 2009. Rutin has therapeutic effect on septic arthritis caused by Candida
albicans. Int. Immunopharmacol. 9, 207–211.
Handa, S.S., Chawla, A.S., Sharma, A.k., 1992. Plants with anti-inflammatory activity.
Fitoterapia LXIII, 3–31.
Hao, H.H., Shao, Z.M., Tang, D.Q., Lu, Q., Chen, X., Yin, X.X., Wu, J., Chen, H., 2012.
Preventive effects of rutin on the development of experimental diabetic
nephropathy in rats. Life Sci. 91, 959–967.
Harborne, J.B., Williams, C.A., 2000. Advances in flavonoid research since 1992.
Phytochemistry 55, 481–504.
Hemwimol, S., Pavasant, P., Shotipruk, A., 2006. Ultrasound-assisted extraction of
anthraquinones from roots of Morinda citrifolia. Ultrason. Sonochem. 13,
543–548.
Hertog, M.G.L., Kromhout, D., Aravanis, C., Blackburn, H., Buzina, R., Fidanza, F.,
Giampaoli, S., Jansen, A., Menotti, A., Nedeljkovic, S., Pekkarinen, M., Simic, B.S.,
Toshima, H., Feskens, E.J.M., Hollman, P.C.H., Katan, M.B., 1995. Flavonoid intake
and long-term risk of coronary heart disease and cancer in the seven countries
study. Arch. Intern. Med. 155, 381–386.
Hollman, P.C.H., de Vries, J.H.M., van Leeuwen, S.D., Mengelers, M.J.B., Katan, M.B.,
1995. Absorption of dietary quercetin glycosides and quercetin in healthy
ileostomy volunteers. Am. J. Clin. Nutr. 62, 1276–1282.
Hollman, P.C.H., Batan, M.B., 1997. Absorption, metabolism and health effects of
dietary flavonoids in man. Biomed. Pharmacother. 51, 305–310.
Hu, Q.H., Wang, C., Li, J.M., Zhang, D.M., Kong, L.D., 2009. Allopurinol, rutin, and
quercetin attenuate hyperuricemia and renal dysfunction in rats induced by
fructose intake: renal organic ion transporter involvement. Am. J. Physiol. 297,
F1080–F1091.
Jacques, R.A., Freitas, L.S., Perez, V.F., Dariva, C., de Oliveira, A.P., de Oliveira, J.V.,
Caramao, E.B., 2007. The use of ultrasound in the extraction of Ilex paraguar-
iensis leaves: a comparison with maceration. Ultrason. Sonochem. 14, 6–12.
Janbaz, K.H., Saeed, S.A., Gilani, A.H., 2002. Protective effect of rutin on paraceta-
mol- and CCl4-induced hepatotoxicity in rodents. Fitoterapia 73, 557–563.
Jiang, P., Burczynski, F., Campbell, C., Pierce, G., Austria, J.A., Briggs, C.J., 2007. Rutin
and flavonoid contents in three buckwheat species Fagopyrum esculentum, F.
tataricum, and F. homotropicum and their protective effects against lipid
peroxidation. Food Res. Int. 40, 356–364.
Jovanovic, S.V., Steenken, S., Tosic, M., Marjanovic, B., Simic, M.G., 1994. Flavonoids
as antioxidants. J. Am. Chem. Soc. 116, 4846–4851.
Jung, M., Ahn, K., Lee, Y., Kim, K., Rhee Paeng, I., Rhee, J., Tae Park, J., Paeng, K., 2001.
Evaluation on the adsorption capabilities of new chemically modified poly-
meric adsorbents with rotoporphyrin IX. J. Chromatogr. A 917, 87–93.
Jung, C.H., Lee, J.Y., Cho, C.H., Kim, C.J., 2007. Anti-asthmatic action of quercetin and
rutin in conscious guinea-pigs challenged with aerosolized ovalbumin. Arch.
Pharm. Res. 30, 1599–1607.
Kalinova, J., Dadakova, E., 2004. Varietal differences of rutin in common buckwheat
(Fagopyrum esculentum Moench) determined by Micellar Electrokinetic Capil-
lary Chromatography. In: Proceedings at the 9th International Symposium on
Buckwheat, Prague 719–722.
Kamalakkannan, N., Stanely Mainzen Prince, P., 2006. The influence of rutin on the
extracellular matrix in streptozotocin-induced diabetic rat kidney. J. Pharm.
Pharmacol. 58, 1091–1098.
Kaufmann, B., Christen, P., 2002. Recent extraction techniques for natural products:
microwave-assisted extraction and pressurised solvent extraction. Phytochem.
Anal. 13, 105–113.
Kauss, T., Moynet, D., Rambert, J., Al-Kharrat, A., Brajot, S., Thiolat, D., Ennemany, R.,
Fawaz, F., Mossalayi, M.D., 2008. Rutoside decreases human macrophage-
derived inflammatory mediators and improves clinical signs in adjuvant-
induced arthritis. Arthritis Res. Ther. 10, R19.
Kayashita, J., Shimaoka, I., Nakajoh, M., Yamazaki, M., Kato, N., 1997. Consumption
of buckwheat protein lowers plasma cholesterol and raises fecal neutral sterols
in cholesterol-fed rats because of its low digestibility. J. Nutr. 127, 1395–1400.
Khan, M.M., Ahmad, A., Ishrat, T., Khuwaja, G., Srivastawa, P., Khan, M.B., Raza, S.S.,
Javed, H., Vaibhav, K., Khan, A., Islam, F., 2009. Rutin protects the neural damage
induced by transient focal ischemia in rats. Brain Res. 1292, 123–135.
Kim, K.H., Lee, K.W., Kim, D.Y., Park, H.H., Kwon, I.B., Lee, H.J., 2005. Optimal
recovery of high-purity rutin crystals from the whole plant of Fagopyrum
esculentum Moench (buckwheat) by extraction, fractionation, and recrystalliza-
tion. Bioresour. Technol. 96, 1709–1712.
Knekt, P., Kumpulainen, J., Jarvinen, R., Rissanen, H., Heliovaara, M., Reunanen, A.,
Hakulinen, T., Aromaa, A., 2002. Flavonoid intake and risk of chronic diseases.
Am. J. Clin. Nutr. 76, 560–568.
Koda, T., Kuroda, Y., Imai, H., 2008. Protective effect of rutin against spatial memory
impairment induced by trimethyltin in rats. Nutr. Res. 28, 629–634.
Koones, H.F., Clifton, N.J., 1948. Extraction of rutin. United States Patent Office
(2,450,555), 5 Oct 1948.
Korkmaz, A., Kolankaya, D., 2010. Protective effect of rutin on the ischemia/
reperfusion induced damage in rat kidney. J. Surg. Res. 164, 309–315.
Kreft, S., Knapp, M., Kreft, I., 1999. Extraction of rutin from buckwheat (Fagopyrum
esculentum Moench) seeds and determination by capillary electrophoresis. J.
Agric. Food Chem. 47, 4649–4652.
Kuhnau, J., 1976. The flavonoids. A class of semi-essential food components: their
role in human nutrition. World Rev. Nutr. Dietetics 24, 117–191.
La Casa, C., Villegas, I., Alarcon de la Lastra, C., Motilva, V., Martin Calero, M.J., 2000.
Evidence for protective and antioxidant properties of rutin, a natural flavone,
against ethanol induced gastric lesions. J. Ethnopharmacol. 71, 45–53.
Landolfi, R., Mower, R.L., Steiner, M., 1984. Modification of platelet function and
arachidonic acid metabolism by bioflavonoids. Structure–activity relations.
Biochem. Pharmacol. 33, 1525–1530.
Lee, A.T., Cerami, A., 1992. Role of glycation in aging. Ann. N. Y. Acad. Sci. 663,
63–70.
Lee, W., Ku, S.K., Bae, J.S., 2012. Barrier protective effects of rutin in LPS-induced
inflammation in vitro and in vivo. Food Chem. Toxicol. 50, 3048–3055.
Levy, G.N., 1997. Prostaglandin H synthases, non-steroidal anti-inflammatory drugs
and colon cancer. FASEB J. 11, 234–247.
Li, F.J., Ning, S.L., Li, Y., Yu, J.Y., Shen, C.D., Duan, G.L., 2012. Optimisation of infrared-
assisted extraction of rutin from crude Flos sophorae Immaturus using response
surface methodology and HPLC analysis. Phytochem. Anal. 23, 292–298.
Li, H., Chen, B., Nie, L., Yao, S., 2004a. Solvent effects on focused microwave assisted
extraction of polyphenolic acids from Eucommia ulmodies. Phytochem. Anal. 15,
306–312.
Li, H., Pordesimo, L., Weiss, J., 2004b. High intensity ultrasound-assisted extraction
of oil from soybeans. Food Res. Int. 37, 731–738.
Macikova, P., Halouzka, V., Hrbac, J., Bartak, P., Skopalova, J., 2012. Electrochemical
behavior and determination of rutin on modified carbon paste electrodes. Sci.
World J. 2012, http://dx.doi.org/10.1100/2012/394756. (Article ID 394756 (9 pp.).
Mahmoud, A.M., 2012. Influence of rutin on biochemical alterations in hyperam-
monemia in rats. Exp. Toxicol. Pathol. 64, 783–789.
Manach, C., Morand, C., Texier, O., 1995. Quercetin metabolites in plasma of rats fed
diets containing rutin or quercetin. J. Nutr. 125, 1911–1922.
Manach, C., Texier, O., Regerat, F., Agullo, G., Demigne, C., Remesy, C., 1996. Dietary
quercetin is recovered in rat plasma as conjugated derivatives of isorhamnetin
and quercetin. J. Nutr. Biochem. 7, 375–380.
Manach, C., Morand, C., Demigne, C., Texier, O., Regerat, F., Remesy, C., 1997.
Bioavailability of rutin and quercetin in rats. FEBS Lett. 409, 12–16.
Michalkiewicz, A., Biesaga, M., Pyrzynska, K., 2008. Solid-phase extraction proce-
dure for determination of phenolic acids and some flavonols in honey. J.
Chromatogr. A 1187, 18–24.
Middleton Jr, E., Kandaswami, C., 1994. In: Harborne, J.B. (Ed.), The Flavonoids:
Advances in Research Since 1986. Chapman and Hall, London, pp. 619–652.
Middleton Jr., E., Kandaswami, C., Theoharides, T.C., 2000. The effects of plant
flavonoids on mammalian cells: implications for inflammation, heart disease,
and cancer. Pharmacol. Rev. 52, 673–751.
Milde, J., Elstner, E.F., Grassmann, J., 2004. Synergistic inhibition of low-density
lipoprotein oxidation by rutin, gamma-terpinene, and ascorbic acid. Phytome-
dicine 11, 105–113.
Montero, J., Mari, M., Colell, A., Morales, A., Basanez, G., Garcia-Ruiz, C., Fernandez-
Checa, J.C., 2010. Cholesterol and peroxidized cardiolipin in mitochondrial
membrane properties, permeabilization and death. Biochim. Biophys. Acta
797, 1217–1224.
 L.S. Chua / Journal of Ethnopharmacology 150 (2013) 805–817
Mosquera, O.M., Correa, Y.M., Buitrago, D.C., Ni, J., 2007. Antioxidant activity of
twenty five plants from Colombian biodiversity. Mem. Inst. Oswaldo Cruz 102,
631–634.
Nagasawa, T., Tabata, N., Ito, Y., Nishizawa, N., 2002. Suppression of early and
advanced glycation by dietary water-soluble rutin derivative in diabetic rats.
Int. Congr. Ser. 1245, 403–405.
Ohnishi, M., Morishita, H., Iwahashi, H., Toda, S., Shirataki, Y., Kimura, M., Kido, R.,
1994. Inhibitory effects of chlorogenic acids on linoleic acid peroxidation and
haemolysis. Phytochemistry 36, 579–583.
Ohsawa, R., Tsutsumi, T., 1995. Improvement of rutin content in buckwheat flour. In:
Matano, T., Ujihara, A., (Eds.), Current Advances in Buckwheat Research
(Proceedings of Sixth International Symposium on Buckwheat at Ina). Shinshu
University Press, Shinshu, pp. 365–372.
Olthof, M.R., Hollman, P.C., Buijsman, M.N., van Amelsvoort, J.M., Katan, M.B., 2003.
Chlorogenic acid, quercetin-3-rutinoside and black tea phenols are extensively
metabolized in humans. J. Nutr. 133, 1806–1814.
Pace-Asciak, C.R., Harm, S., Diamandis, E.P., Soleas, G., Goldberg, D.M., 1995. The red
wine phenolics trans-resveratrol and quercetin block human platelet aggrega-
tion and eicosanoid synthesis: implications for protection against coronary
heart disease. Clin. Chim. Acta 235, 207–219.
Paganga, G., Miller, N., Rice-Evans, C.A., 1999. The polyphenolic content of fruit and
vegetables and their antioxidant activities. What does a serving constitute?
Free Radical Res. 30, 153–162.
Paniwnyk, L., Beaufoy, E., Lorimer, J.P., Mason, T.J., 2001. The extraction of rutin
from flower buds of Sophora japonica. Ultrason. Sonochem. 8, 299–301.
Pare, J.R.J., Belanger, J.M.R., Stafford, S.S., 1994. Microwave-assisted process (MAPTM): a
new tool for the analytical laboratory. Trends Anal. Chem. 13, 176–184.
Picot, D., Loll, P.J., Garavito, R.M., 1994. The X-ray crystal structure of the membrane
protein prostaglandin H2 synthase-1. Nature 367, 243–249.
Pinelo, M., Rubilar, M., Jerez, M., Sineiro, J., Nunez, M.J., 2005. Effect of solvent,
temperature, and solvent-to-solid ratio on the total phenolic content and
antiradical activity of extracts from different components of grape pomace. J.
Agric. Food Chem. 53, 2111–2117.
Pu, F., Mishima, K., Irie, K., Motohashi, K., Tanaka, Y., Orito, K., Egawa, T., Kitamura,
Y., Egashira, N., Iwasaki, K., Fujiwara, M., 2007. Neuroprotective effects of
quercetin and rutin on spatial memory impairment in an 8-arm radial maze
task and neuronal death induced by repeated cerebral ischemia in rats. J.
Pharmacol. Sci. 104, 329–334.
Ramanathan, R., Das, W.P., Tan, C.H., 1993. Inhibitory effects of 2-hydroxy chalcone and
other flavonoids on human cancer cell-proliferation. Int. J. Oncol. 3, 115–119.
Ren, W., Qiao, Z., Wang, H., Zhu, L., Zhang, L., 2003. Flavonoids: promising
anticancer agents. Med. Res. Rev. 23, 519–534.
Reynolds, J.E.F., 1996. Martindale-The Extra Pharmacopoeia, 31st ed. The Royal
Pharmaceutical Society, Council of the Royal Pharmaceutical Society of Great
Britain, London, pp. 1679–1680.
Rimm, E.B., Katan, M.B., Ascherio, A., Stamper, M.J., Willett, W.C., 1996. Relation
between intake of flavonoids and risk for coronary heart disease in male health
professionals. Ann. Intern. Med. 125, 384–389.
Sando, C.E., Lloyd, J.U., 1924. The isolation and identification of rutin from the
flowers of elder (Sambucus canadensis L.). J. Biol. Chem., 737–745.
Sawai, Y., Kohsaka, K., Nishiyama, Y., Ando, K., 1987. Serum concentrations of
rutoside metabolites after oral administration of a rutoside formulation to
humans. Arzneimittelforschung 37, 729–732.
Schinor, E.C., Salvador, M.J., Turatti, I.C.C., Zucchi, O.L.A.D., Dias, D.A., 2004.
Comparison of classical and ultrasound assisted extractions of steroids and
triterpenoids from three Chresta spp. Ultrason. Sonochem. 11, 415–421.
Schneider, H., Simmering, R., Hartmann, L., Pforte, H., Blaut, M., 2000. Degradation
of quercetin-3-glucoside in gnotobiotic rats associated with human intestinal
bacteria. J. Appl. Microbiol. 89, 1027–1037.
Scordino, M., Mauro, A.D., Passerini, A., Maccarone, E., 2003. Adsorption of
flavonoids on resins: hesperidin. J. Agric. Food Chem. 51, 6998–7004.
Shahrokhian, S., Ghalkhani, M., Amini, M.K., 2009. Application of carbon-paste
electrode modified with iron phthalocyanine for voltammetric determination
of epinephrine in the presence of ascorbic acid and uric acid. Sensors Actuators
B 137, 669–675.
Sharma, S., Sahu, D., Das, H.R., Sharma, D., 2011. Amelioration of collagen-induced
arthritis by Salix nigra bark extract via suppression of pro-inflammatory
cytokines and oxidative stress. Food Chem. Toxicol. 49, 3395–3406.
Shen, S.C., Lee, W.R., Lin, H.Y., Huang, H.C., Ko, C.H., Yang, L.L., Chen, Y.C., 2002. In
vitro and in vivo inhibitory activities of rutin, wogonin, and quercetin on
lipopolysaccharide induced nitric oxide and prostaglandin E2 production. Eur. J.
Pharmacol. 446, 187–194.
Silva, E.M., Pompeu, D.R., Larondelle, Y., Rogez, H., 2007. Optimisation of the
adsorption of polyphenols from Inga edulis leaves on macroporous resins using
an experimental design methodology. Sep. Purif. Technol. 53, 274–280.
Stanley Mainzen Prince, P., Kamalakkannan, N., 2006. Rutin improves glucose
homeostasis in streptozotocin diabetic tissues by altering glycolytic and
gluconeogenic enzymes. J. Biochem. Mol. Toxicol. 20, 96–102.
Stavarache, C., Vinatoru, M., Maeda, Y., 2007. Aspects of ultrasonically assisted
transesterification of various vegetable oils with methanol. Ultrason. Sono-
chem. 14, 380–386.
Suman, R., Saffron, A.W., 2008. Phytoestrogens oestrogen synthesis and breast
cancer. J. Steroid Biochem. Mol. Biol. 3, 186–195.
Sun, W.Q., Sheng, J.F., 1998. Handbook of Natural Active Constituents. Chinese
Medicinal Science and Technology Press, Beijing, pp. 2240–2316.
817
Suzuki, T., Honda, Y., Mukasa, Y., 2005. Effects of UV-B radiation, cold and
desiccation stress on rutin concentration and rutin glucosidase activity in
tartary buckwheat (Fagopyrum tataricum) leaves. Plant Sci. 168, 1303–1307.
Takahama, U., Hirota, S., 2000. Deglucosidase of quercetin glucosides to the
aglycone and formation of antifungal agents by peroxidase-dependent oxida-
tion of quercetin on browing of onion scales. Plant Cell Physiol. 41, 1021–1029.
Teresita, G., Ester, R.A., Osvaldo, J.A., Eugenia, P.L., 2001. Anti-inflammatory proper-
ties of plant flavonoids. Effects of rutin, quercetin and hesperidin on adjuvant
arthritis in rat. II Farmaco 56, 683–687.
Tham, D.M., Gardner, C.D., Haskel, W.L., 1998. Potential health benefits of dietary
phytoestrogens: a review of the clinical, epidemiological, and mechanistic
evidence. J. Clin. Endocrinol. Metab. 83, 2223–2235.
Toma, M., Vinatoru, M., Paniwnyk, L., Mason, T.J., 2001. Investigation of the effects
of ultrasound on vegetal tissues during solvent extraction. Ultrason. Sonochem.
8, 137–142.
Tongjaroenbuangam, W., Ruksee, N., Chantiratikul, P., Pakdeenarong, N., Kongbun-
tad, W., Govitrapong, P., 2011. Neuroprotective effects of quercetin, rutin and
okra (Abelmoschus esculentus Linn.) in dexamethasone-treated mice. Neuro-
chem. Int. 59, 677–685.
Toyokuni, S., 1996. Iron-induced carcinogenesis: the role of redox regulation. Free
Radical Biol. Med. 20, 553–566.
Tucker, H.A., 1981. Physiological control of mammary growth, lactogenesis and
lactation. J. Dairy Sci. 64, 1403–1421.
Umar, S., Mishra, N.K., Pal, K., Sajad, M., Neha, Ansari, M.M., Ahmad, S., Katiyar, C.K.,
Khan, H.A, 2012. Protective effect of rutin in attenuation of collagen-induced
arthritis in Wistar rat by inhibiting inflammation and oxidative stress. Indian J.
Rheumatol. 7, 191–198.
Ushida, Y., Matsui, T., Tanaka, M., Matsumoto, K., Hosoyama, H., Mitomi, A.,
Sagesaka, Y., Kakuda, T., 2008. Endothelium-dependent vasorelaxation effect
of rutin-free tartary buckwheat extract in isolated rat thoracic aorta. J. Nutr.
Biochem. 19, 700–707.
Wang, J., Wu, F.A., Zhao, H., Liu, L., Wu, Q.S., 2008. Isolation of flavonoids from
mulberry (Morus alba L.) leaves with macroporous resins. Afr. J. Biotechnol. 7,
2147–2155.
Webster, R.P., Gawde, M.D., Bhattacharya, R.K., 1996. Protective effect of rutin, a
flavonol glycoside, on the carcinogen-induced DNA damage and repair enzymes
in rats. Cancer Lett. 109, 185–191.
Williams, C.A., Harborne, J.B., 1977. The leaf flavonoids of the Zingiberales. Biochem.
Syst. Ecol. 5, 221–229.
Winter, J., Moore, L.H., Dowell Jr, V.R., Bokkenheuser, V.D., 1989. C-Ring cleavage of
flavonoids by human intestinal bacteria. Appl. Environ. Microbiol. 55, 1203–1208.
Wong, S.C.Y., Fukuchi, M., Melnyk, P., Rodger, I., Giaid, A., 1998. Induction of
cyclooxygenase-2 and activation of nuclear factor kB in myocardium of patients
with congestive heart failure. Circulation 98, 100–103.
Wu, H., Chen, M., Fan, Y., Elsebaei, F., Zhu, Y., 2012. Determination of rutin and quercetin
in Chinese herbal medicine by ionic liquid-based pressurized liquid extraction–
liquid chromatography–chemiluminescence detection. Talanta 88, 222–229.
Van der Logt, E.M., Roelofs, H.M., Nagengast, F.M., Peters, W.H., 2003. Induction of
rat hepatic and intestinal UDP glucuronosyltransferases by naturally occurring
dietary anticarcinogens. Carcinogenesis 24, 1651–1656.
Van Rantwijk, F., Sheldon, R.A., 2007. Biocatalysis in ionic liquids. Chem. Rev. 107,
2757–2785.
Vinatoru, M., 2001. An overview of the ultrasonically assisted extraction of
bioactive principles from herbs. Ultrason. Sonochem. 8, 303–317.
Vishwanath, V., Frank, K.E., Elmets, C.A., Dauchot, P.J., Monnier, V.M., 1986.
Glycation of skin collagen in type I diabetes mellitus. Correlation with long-
term complications. Diabetes 35, 916–921.
Xie, J., Shi, L., Zhu, X., Wang, P., Zhao, Y., Su, W., 2011. Mechanochemical-assisted
efficient extraction of rutin from Hibiscus mutabilis L. Innovative Food Sci.
Emerging Technol. 12, 146–152.
Yang, J., Juan Guo, J., Yuan, J., 2008. In vitro antioxidant properties of rutin. LWT-
Food Sci. Technol. 41, 1060–1066.
Yang, Y., Zhang, F., 2008. Ultrasound-assisted extraction of rutin and quercetin from
Euonymus alatus (Thunb.) Sieb. Ultrason. Sonochem. 15, 308–313.
Yoon, S.Y., Choi, W.J., Park, J.M., Yang, J.W., 1997. Selective adsorption of flavonoid
compounds from the leaf extract of Ginkgo biloba L. Biotechnol. Tech. 11,
553–556.
Zeng, H., Wang, Y., Kong, J., Nie, C., Yuan, Y., 2010. Ionic liquid-based microwave-
assisted extraction of rutin from Chinese medicinal plants. Talanta 83, 582–590.
Zeng, H., Wang, Y., Liu, X., Kong, J., Nie, C., 2012. Preparation of molecular imprinted
polymers using bi-functional monomer and bi-crosslinker for soild-phase
extraction of rutin. Talanta 93, 172–181.
Zhang, F., Yang, Y., Su, P., Guo, Z.K., 2009. Microwave-assisted extraction of rutin and
quercetin from the stalks of Euonymus alatus (Thunb.) Sieb. Phytochem. Anal.
20, 33–37.
Zhang, Y., Li, S.F., Wu, X.W., 2008. Pressurized liquid extraction of flavonoids from
Houttuynia cordata Thunb. Sep. Purif. Technol. 58, 305–310.
Zhao, Y., Bao, C., Mason, T.J., 1991. A study of the isolation of effective compositions
from traditional Chinese medicinal plants. In: Ultrasonics International 91
Proceedings. Butterworths, pp. 87–90.
Zhu, F., Cai, Y.Z., Sun, M., Corke, H., 2008. Influence of Amaranthus betacyanin
pigments on the physical properties and color of wheat flours. J. Agric. Food
Chem. 56, 8212–8217.