
Secondary Metabolites of Miconia rubiginosa

Juliana Rodrigues,1 Daniel Rinaldo,1 Marcelo Aparecido da Silva,2

Lourdes Campaner dos Santos,1 and Wagner Vilegas1

1Department of Organic Chemistry, Institute of Chemistry, São Paulo State University, Araraquara, São Paulo, Brazil.
2Department of Pharmacy, Federal University of Alfenas, Alfenas, Minas Gerais, Brazil.

ABSTRACT The Miconia genus is the most representative of the Melastomataceae family, and some species are commonly

used in Brazilian folk medicine as anti-inflammatory agents. In this work we investigated the leaves from Miconia rubiginosa

(Bonpl.) DC, using high-speed countercurrent chromatography, which yielded 11 substances (eight flavonoids, gallic acid,

casuarictin, and schizandriside). Identification was achieved using nuclear magnetic resonance spectroscopy and high-

performance liquid chromatography–circular dichroism–diode array detection analyses.

KEY WORDS: � circular dichroism � high-speed countercurrent chromatography � Melastomataceae

INTRODUCTION

M iconia is a genus of approximately 1,000 species1

occurring in tropical America.2,3 The genus belongs to
the pantropical Melastomataceae family with more than 166
genera that include about 4,300 species.2 Many of these
plants are used as medicine by people living in the Cerrado
area.4 Miconia infusions and extracts and compounds
isolated from members of the genus have demonstrated an-
timicrobial,5 antibiotic, antitumoral, analgesic, and antima-
larial activities.6,7

Previous phytochemical investigations of Miconia spe-
cies have resulted in the isolation of triterpenes,8,9 flavo-
noids,10,11 and quinones.12

The isolation of the compounds present in a crude extract
is often performed by repeated processes based on adsorp-
tion column chromatography. In our case, we investigate
mainly polar extracts containing glycosides, which can be
irreversibly adsorbed on the stationary phase or even be
degraded because of the catalytic activity of some solid
supports. The isolated compounds are subjected to in vivo
pharmacological tests, which require fast separation and
milligram to gram quantities of pure compounds. High-
speed countercurrent chromatography (HSCCC) uses no
solid support and has a greater maximum capacity, excellent
sample recovery, efficient separation of closely related
natural products, and a wider choice of solvent systems
compared with conventional high-performance liquid
chromatography (HPLC).

A prerequisite for the full understanding of molecular
events involved in the biological activity of chiral molecules
is the knowledge of their absolute configuration.13 The
pharmacological and pharmacodynamic properties of bio-
logically active chiral compounds are often strictly related
to their stereochemistry. Enantiomers usually differ in bio-
logical activities because of their disparate interactions with
enzymes and other naturally occurring chiral molecules.14

HPLC with circular dichroism (CD) detection is an excellent
method for the separation and detection of these chiral
compounds.

There are no reports concerning the chirality of molecules
isolated from Miconia species; therefore, the scope of our
analysis was to search for an efficient method to isolate in
preparative scale the secondary metabolites contained in the
infusion of Miconia rubiginosa (Bonpl.) DC and to establish
the absolute configurations of the chiral isolates.

MATERIALS AND METHODS

Reagents

All solvents used for HSCCC were of analytical reagent
grade from Merck (Darmstadt, Germany). The solvents used
for HPLC were of analytical grade from J.T. Baker (Phil-
lipsburg, NJ, USA). Water was nanopure quality, obtained
from a Milli-Q� purification system (Millipore Corp.,
Bedford, MA, USA).

Preparation of crude sample and sample solution

Aerial parts of M. rubiginosa were collected in March
2005 at Palmeiras da Serra, Pratânia, SP, Brazil, and au-
thenticated by Dr. Luiz Fernando Rolim de Almeida from
the Botany Institute, State University of São Paulo, Botucatu,

Manuscript received 16 June 2010. Revision accepted 17 September 2010.

Address correspondence to: Ms. Juliana Rodrigues, Department of Organic Chemistry,
Institute of Chemistry, São Paulo State University, P.O. Box 355, Prof. Francisco Degni
Street s=n, CEP 14800-900, Araraquara, SP, Brazil, E-mail: julirodr@iq.unesp.br

JOURNAL OF MEDICINAL FOOD
J Med Food 14 (7=8) 2011, 834–839
# Mary Ann Liebert, Inc. and Korean Society of Food Science and Nutrition
DOI: 10.1089=jmf.2010.0157

834


SP, Brazil. A voucher specimen (number 25376) was de-
posited in the Irina Delanova Gemtchujnicov Herbarium of
the Biosciences Institute, State University of São Paulo.

The dried leaves of M. rubiginosa (100 g) were separated,
powdered, and extracted at 70�C for 10 minutes in 1 L of
water, yielding an infusion.

Because of the presence of tannins, the infusion (1 L) was
partitioned with ethyl acetate (3 · 500 mL), with the poly-
meric tannins remaining in the aqueous phase and the sec-
ondary metabolites transferred to the ethyl acetate phase.
The ethyl acetate fraction was evaporated at 35�C under re-
duced pressure, yielding crude extract (1.5 g, 1.5% yield).
The ethyl acetate fraction (1.5 g) was dissolved in a mixture
consisting of 20 mL lower phase + 20 mL upper phase of the
solvent system ethyl acetate=n-propanol=H2O (14:0.8:8 by
volume).

HSCCC

The preparative HSCCC instrument used in this study was
from P.C. Inc. (Potomac, MD, USA). It was equipped with a
multiplayer with two coils of 1.68-mm (i.d.) polytetra-
fluoroethylene tubing of approximately 80 and 240 mL, re-
spectively, with a total capacity of 320 mL. The b value
varied from 0.5 at the internal to 0.85 at the external terminal,
and the revolution radius was 10 cm (b = r=R, where r is the
distance from the coil to the holder, and R is the revolution
radius or the distance between the holder axis and the central
shaft). The flow rate was controlled with a Waters (Milford,
MA, USA) model 4000 constant-flow pump. The sample was
injected with a injection module (P.C. Inc.) with a 20-mL
sample injection loop. The coiled column was filled with the
stationary phase (lower phase), then the apparatus was rotated
forward at 850 rpm, and the mobile phase (upper phase) was
pumped into the column in a head-to-tail direction at a flow
rate of 1.0 mL=minute. After the mobile phase front emerged
and the hydrodynamic equilibrium was established in the
column, about 20 mL of the sample solution containing 0.75 g
of the ethyl acetate fraction was injected through the injection
module at a flow rate of 1.0 mL=minute. Sixty fractions of
5 mL each were collected with a Redifrac automated fraction
collector (Pharmacia, Uppsala, Sweden), in approximately 5
hours. After thin-layer chromatography (TLC) analyses,
fractions with similar retardation factor (RF) were combined.
The procedure was repeated with the remaining 0.75-g ethyl
acetate fraction.

Preparation of the two-phase solvent system

The solvent system, composed of ethyl acetate=n-propanol=
H2O (14:0.8:8 by volume), was thoroughly equilibrated
overnight in a separator funnel at room temperature, and the
two phases were separated shortly before use.

TLC and HPLC analyses

Aliquots of the ethyl acetate and the collected fractions
were analyzed on glass-backed silica gel TLC (20 · 20 cm,
Aldrich, Milwaukee, WI, USA), with the mobile phase

being CHCl3=methanol=n-propanol=H2O (5:6:1:4 by vol-
ume). The TLC plates were evaluated under ultraviolet (UV)
light (254 nm) and derivatized with anisaldehyde=H2SO4 so-
lution.15 The infusion and the isolated metabolites were ad-
ditionally analyzed using a Jasco� (Tokyo, Japan) HPLC
system equipped with a PU-2089 Plus pump, a MD-2010 Plus
photodiode array detector, a CD-2095 Plus CD detector, an
AS-2055 Plus autosampler, and a C18 column (250 · 4.60 mm
i.d.; particle size, 5 lm; Luna, Phenomenex, Torrance, CA,
USA), and a Phenomenex security guard (4.0 · 2.0 mm i.d.).
EZChrom Elite� version 3.1.7 software (Agilent Technolo-
gies, Palo Alto, CA, USA) was used for control of the ana-
lytical system, data collection, and processing. The mobile
phase was acetonitrile + trifluoroacetic acid (0.05%) (eluent
A) and H2O + 0.05% trifluoroacetic acid (eluent B), with a
linear gradient elution of A:B 13:87 (vol=vol) to A:B 23:77
(vol=vol) in 40 minutes and then to 100% A in 45 minutes at a
flow rate of 1.0 mL=minute, and the effluent was monitored at
254 nm. The infusion was applied to a solid-phase extraction
cartridge (20 mg, Sep Pak C18, Waters), which was pre-
conditioned with 10 mL of HPLC-grade methanol and 10 mL
of water (18 mO$cm). A suitable amount of sample (to load
20 mg of infusion=g of solid phase) was loaded onto the C18

cartridge, eluted with 5 mL of H2O=methanol (8:2, fraction 1),
5 mL of H2O=methanol (1:1, fraction 2), and 5 mL of meth-
anol (100%, fraction 3), and dried using a stream of N2. The
residues of the three fractions were redissolved in 2 mL of
H2O=methanol (8:2, vol=vol) and filtered through a mem-
brane polytetrafluoroethylene filter (pore size, 0.45 lm; Mill-
ex, Millipore), and 20lL of this filtrate was subjected to
HPLC-UV-diode array detection (DAD) analysis.

The chiral study was performed with a polysaccharide-
derived chiral stationary phase column (Chiralcel OD-RH�;
particle size, 5 lm; 250 · 4.6 mm) protected by a Chiralcel
OD-RH guard column (particle size, 5 lm; 10 · 4.0 mm)
(Daicel Chemical Industries, Tokyo). The CD spectra were
scanned between 220 and 420 nm for the resolved enantio-
mers using the online CD detector. The spectra were average
computed over three instrumental scans, and the intensities
are presented in terms of ellipticity values.

Structural identification of the compounds

Nuclear magnetic resonance (NMR) spectra (one- and
two-dimensional) were obtained in dimethyl sulfoxide
(DMSO)-d6 using a Varian (Palo Alto) INOVA 500 spec-
trometer operating at 500 MHz for 1H and 125 MHz for 13C.

Spectroscopic data

Compound 9 was a white amorphous powder of compo-
sition C15H14O6 with kmax (in methanol) of 214 and 279 nm.
By 1H-NMR (DMSO-d6): 2.50 (dd, J = 4.5; 16.0 Hz, 1H, H-
4b), 2.67 (dd, J = 4.5; 16.0 Hz, 1H, H-4a), 4.02 (s, 1H, H-3),
4.74 (s, 1H, H-2), 5.73 (d, J = 2 Hz, 1H, H-6), 5.89 (d,
J = 2 Hz, 1H, H-8), 6.66 (m, 1H, H-50), 6.67 (m, 1H, H-60),
6.89 (d, J = 1.5 Hz, 1H, H-20).

Compound 10 was a colorless amorphous powder of
composition C41H28O26 with kmax (in methanol) of 223 and

SECONDARY METABOLITES OF M. RUBIGINOSA 835


280 nm. By 1H-NMR (DMSO-d6): 3.84 (d, J = 13 Hz, 1H, H-
6a), 4.58 (dd, J = 7.0; 9.0 Hz, 1H, H-5), 4.99 (t, J = 10 Hz,
1H, H-4), 5.00 (t, J = 10 Hz, 1H, H-2), 5.12 (dd, J = 6.5;
13.0 Hz, 1H, H-6b), 5.49 (t, J = 9.0 Hz, 1H, H-3), 6.27 (d,
J = 8.5 Hz, 1H, H-1 anom), 6.17, 6.25, 6.38, 6.44 (s, each
1H, HHDP-H), 7.02 (s, 2H, galloyl H).

Compound 11 was colorless needles of composition
C25H32O10 with kmax (in methanol) of 207 and 284 nm. By
1H-NMR (DMSO-d6): 1.72 (t, J = 10.5 Hz, 1H, H-80), 1.89
(tt, J = 3.5; 10.5 Hz, 1H, H-8), 2.73 (d, J = 6.5 Hz, 2H, H-7),
2.98 (m, 1H, H-500), 2.99 (m, 1H, H-90), 2.99 (m, 1H, H-200),
3.09 (dt, J = 9.0 Hz, 1H, H-300), 3.27 (m, 1H, H-400), 3.47 (m,
1H, H-9), 3.58 (dd, J = 4.0; 10.0 Hz, 1H, H-9), 3.66 (dd,
J = 5.5; 11.5 Hz, 1H, H-500), 3.70 (s, 1H, OCH3), 3.72 (s, 1H,
OCH3), 3.86 (dd, J = 2.5; 9.5 Hz, 1H, H-90), 3.93 (d,
J = 7.5 Hz, 1H, H-100), 4.03 (d, J = 10.5 Hz, 1H, H-70), 6.10
(s, 1H, H-3), 6.49 (dd, J = 1.5; 8.0 Hz, 1H, H-60), 6.62 (s, 1H,
H-6), 6.70 (d, J = 8.0 Hz, 1H, H-50), 6.80 (d, J = 1.5 Hz, 1H,
H-20).

RESULTS

A series of experiments was performed to determine a
suitable two-phase solvent system for HSCCC. Small
amounts of the ethyl acetate fraction were dissolved in test
tubes containing each of the following solvent systems:
ethyl acetate=n-propanol=H2O (14:0.8:8 by volume), ethyl
acetate=n-butanol=H2O (14:0.4:8 by volume), and hexane=
ethyl acetate=methanol=H2O (1:1:1:1 by volume). The test
tubes were shaken, and the system was allowed to equili-
brate between the two phases. An aliquot of each phase was
spotted on a TLC plate (CHCl3=methanol=n-propanol=H2O
[5:6:1:4 by volume]), visualized under UV light (254 nm),
and developed with anisaldehyde=H2SO4 solution. The two-
phase solvent system was selected according to the partition
coefficients (K) of the target components. The K values were
determined by TLC analysis according to Conway.16 The
best result was obtained with the mixture of ethyl acetate=
n-propanol=H2O (14:0.8:8 by volume), with the substances

FIG. 1. Secondary metabolites of M.
rubiginosa.

836 RODRIGUES ET AL.

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almost equally distributed between the two phases. The
relatively high proportion of ethyl acetate in the solvent
mixture and the RF values of the compounds indicate the
medium polarity of the metabolites of M. rubiginosa. Thus
the upper phase was chosen as the mobile phase, and the
lower phase was used as the stationary phase for the HSCCC
separation of the ethyl acetate fraction of the infusion of M.
rubiginosa. This choice has the additional advantage that the
upper phase consists largely of the volatile ethyl acetate,
readily eliminated by evaporation.

Under the conditions described, the retention of the sta-
tionary phase in the HSCCC was 87%. This method led to
the isolation of the compounds in approximately 3.5 hours,
and the whole process was completed in 5 hours. Spectro-
scopic analyses allowed identification of 11 main compounds,
determined by comparison with literature data: quercetin-3-O-
a-rhamnopyranoside (1),17 quercetin-3-O-b-arabinofuranoside
(2),18 quercetin-3-O-a-arabinopyranoside (3),19 quercetin-3-O-
b-arabinopyranoside (4),19 quercetin-3-O-b-galactopyranoside
(5),18 quercetin-3-O-a-rhamnopyranosil-(1/4)-O-b-galacto-
pyranoside (6),19 kaempferol-3-O-b-galactopyranoside (7),19

gallic acid (8),19,20 (–)-epicatechin (9),4 casuarictin (10),21 and
schizandriside (11)22 (Fig. 1).

The identity and purity of the isolated substances were
verified by HPLC-CD-DAD analyses using authentic sam-
ples from a collection of our laboratory. All substances were
obtained with purity > 95%.

Figure 2 shows the HPLC chromatographic analysis of
fraction 2 of the infusion of M. rubiginosa aerial parts. The
peaks were identified by co-injection with purified metab-
olites.

The compounds isolated—(–)-epicatechin (9), casuarictin
(10), and schizandriside (11)—were analyzed by HPLC-
CD-DAD using a chiral column (OD-RH) with metha-

nol=H2O (4:6, vol=vol) as eluent. The chromatograms
showed only one peak, indicating that all the substances
were present in enantiomerically pure forms in this species.
The CD spectra were recorded for the establishment of the
absolute configuration of these substances (Fig. 3).

DISCUSSION

Several classes of natural products have been isolated by
HSCCC, including flavonoids.23,24 Our group reported the
isolation of naphthopyranone glycosides from Paepalanthus
microphyllus (Family Eriocaulaceae),25 quercetin glyco-
sides and the biflavonoid amentoflavone from Byrsonima
crassa (Family Malpighiaceae),26 the flavones 6-methoxy-
luteolin-7-O-b-d-allopyranoside and 6-methoxyapigenin-7-
O-b-d-allopyranoside from Eriocaulon ligulatum (Family
Eriocaulaceae),27 and quercetin-3-O-a-l-rhamnopyranoside
and myricetin-3-O-a-l-rhamnopyranoside from Davilla el-
liptica (Family Dilleniaceae).28

The separation mechanism of HSCCC is based mainly on
the differential solubility of the compound between the two
immiscible phases, one flowing along the other, allowing the
fast and efficient isolation of several compounds without
decomposition. HSCCC, being a support-free liquid–liquid
partition chromatographic technique, eliminates irrevers-
ible adsorption of the sample onto the solid support, and

FIG. 2. Chromatogram of the infusion of M. rubiginosa by high-
performance liquid chromatography analysis: quercetin-3-O-
a-rhamnopyranoside (1), quercetin-3-O-b-arabinofuranoside (2),
quercetin-3-O-a-arabinopyranoside (3), quercetin-3-O-b-arabinopyr-
anoside (4), quercetin-3-O-b-galactopyranoside (5), and quercetin-
3-O-a-rhamnopyranosil-(1/4)-O-b-galactopyranoside (6). Eluent A,
acetonitrile + 0.05% trifluoroacetic acid; eluent B, H2O + 0.05% tri-
fluoroacetic acid. Method: 13–23% B in A (40 minutes); 23–100% B in
A (45 minutes); flow rate, 1.0 mL=minute; injection volume, 20 lL;
Phenomenex Luna C18 column; detection at 254 nm. *Might be inverted.

FIG. 3. Circular dichroism (CD) spectra of (a) (–)-epicatechin (9),
(b) casuarictin (10), and (c) schizandriside (11). mdeg, ellipticity
value.

SECONDARY METABOLITES OF M. RUBIGINOSA 837

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therefore this method has been widely used for the prepar-
ative separation of natural products.29,30

Despite the similar polarities of the compounds, they
were well separated, with the separation of the quercetin and
kaempferol glycosides dependent on the total number of the
hydroxyl groups. Our results showed that the conditions
used provided an efficient method for the separation of the
metabolites from M. rubiginosa.

Catechins are an important class of secondary metabolites.
They show strong antioxidant activity and have various
health-related activities, such as anticarcinogenic, anti-
allergic, anti-atherogenic, antibacterial, and antiviral activi-
ties.31 These biological activities are significantly influenced
by stereochemical differences, thus justifying the determi-
nation of the chirality of these metabolites.

Catechin derivatives have two stereocenters, resulting in
four diastereomers, These substances have two aromatic
chromophores, namely, the benzene A- and B-rings, with
the absorption bands of these chromophores found at ap-
proximately 280 and 240 nm.32 The positive Cotton effect
in the 240 nm region (La transition, chromophore B) and
the negative Cotton effect in the 280 nm (Lb transition,
chromophore A) regions indicate (2R,3R) absolute config-
uration, elucidating 9 as (–)-epicatechin (Fig. 3). This is the
first report of the isolation of catechins from plants of the
Melastomataceae family.

Casuarictin (10) is an example of a hydrolyzable tannin.
Plants with high tannin content are used in traditional
medicine to treat various diseases such as diarrhea, hyper-
tension, rheumatism, bleeding, wounds, burns, stomach
problems, kidney problems, and inflammatory processes.33

Several studies have shown the presence of hydrolyzable
tannins in species of the Melastomataceae family.34,35

Casuarictin (10) (which is a monomeric b-glucosyl ella-
gitannin) presents atropisomerism due to the restricted ro-
tation of the biphenyl bond of the hexahydroxydiphenoyl
(HHDP) group. Casuarictin (10) displayed three Cotton ef-
fects, around 235, 265, and 285 nm. These Cotton effects are
associated with the biphenyl conjugation bands in the UV
spectra. The Cotton effect at 235 nm is diagnostic for the
absolute configuration of the HHDP group [positive for the
(S)- and negative for the (R)-configuration].21

The CD spectra of 10 exhibited a positive Cotton effect at
235 nm, thus indicating the (S)-configuration for HHDP
groups.

Schizandriside (11) is an aryltetralin-type glycosylated
lignan. This compound exhibited moderate antioxidant ac-
tivity22 and moderate antibacterial activity against Escher-
ichia coli, Staphylococcus aureus, Micrococcus luteus, and
Bacillus cereus using the paper disk method.36

The absolute structure of 11 was established based on CD
spectroscopic evidence. Klyne et al.37 proposed a CD em-
pirical rule that states that all 70a-aryl derivatives in 70-
aryltetralin-type lignans give a positive Cotton effect at
280–290 nm, whereas all 70b-aryl groups display a negative
Cotton effect. The CD spectrum of 11 afforded a negatively
signed maximum at 284 nm, indicating 70b (70S) configu-
ration. The large 3J70,80 and 3J8,80 coupling constants

(J = 11.0 Hz) established the absolute configuration as
(70S,80R,8R).

The results of our studies clearly demonstrate the poten-
tial of HSCCC for the preparative isolation of secondary
metabolites from the leaves of M. rubiginosa. In particular,
preparative HSCCC, with its speedy separation and mini-
mum solvent consumption, offers an efficient method for the
separation and purification of natural products.

The compounds 1, 8, and 9 were previously isolated from
the Miconia cabucu species.10 The others were isolated for
the first time from Miconia species. There has been no report
about the optical study of metabolites isolated from species
of the Melastomataceae family. The HPLC-CD-DAD ana-
lyses showed that all the chiral substances isolated from
M. rubiginosa were presented in enantiomerically pure
forms. Their absolute configurations were established as
(–)-epicatechin, casuarictin [(S)-HHDP], and (7S,8R,80R)-
schizandriside, respectively.

ACKNOWLEDGMENTS

We thank the Fundação de Amparo à Pesquisa do Estado
de São Paulo for funding from the Biota-Fapesp Program
and the Conselho Nacional de Desenvolvimento Cientı́fico
Tecnológico for a grant to W.V. We also thank CAPES for a
fellowship to J.R. and M.A.S.

AUTHOR DISCLOSURE STATEMENT

No competing financial interests exist.

REFERENCES

1. Martins AB, Semir J, Goldenberg R, Martins E: O gênero Mi-

conia Ruiz & Pav. no Estado de São Paulo. Acta Bot Bras

1996;10:267–316.

2. Renner SS: Phylogeny and classification of the Melastomataceae

and Memecylaceae. Nord J Bot 1993;13:519–540.

3. Judd WS, Skean JD Jr: Phylogeny and classification of the

Melastomataceae and Memecylaceae. Bull Florida Mus Nat Hist

1991;36:25–84.

4. Almeida SCX, Lemos TLG, Silveira ER, Pessoa ODL: [Volatile

and non-volatile chemical constituents of Cochlospermun viti-

folium (Willdenow) Sprengel]. Quim Nova 2005;28:57–60.

5. Rodrigues J, Michelin DC, Rinaldo D, Zocolo GJ, Dos Santos

LC, Vilegas W, Salgado HRN: Antimicrobial activity of Miconia

species (Melastomataceae). J Med Food 2008;11:120–126.

6. Hasrat JA, De Backer JP, Valquelin G, Vlietinck AJ: Medicinal

plants in Suriname: screening of plants extracts for recepto-

binding activity. Phytomedicine 1997;4:56–65.

7. Cunha WR, Martins C, Ferreira DS, Crotti AEM, Lopes NP,

Albuquerque S: In vitro trypanocidal activity of triterpenes from

Miconia species. Planta Med 2003;69:468–470.

8. Chan WR, Sheppard V, Medford KA, Tinto WF: Triterpenes

from Miconia stenostachya. J Nat Prod 1992;55:963–966.

9. Macari PAT, Emerenciano VP, Ferreira ZMGS: [Identification of

triterpenes from Miconia albicans through analysis by micro-

computer]. Quim Nova 1990;13:260–262.

10. Rodrigues J, Rinaldo D, Dos Santos LC, Vilegas W: An unusual

C6–C60 0 linked flavonoid of Miconia cabucu (Melastomataceae).

Phytochemistry 2007;68:1781–1784.

838 RODRIGUES ET AL.


11. Li XC, Jacob MR, Pasco DS, ElSohly HN, Nimrod AC, Walker

LA, Clark AM: Phenolic compounds from Miconia myriantha

inhibiting Candida aspartic proteases. J Nat Prod 2001;64:1282–

1285.

12. Bernays E, Lupi A, Bettolo RM, Mastrofrancesco C, Tagliatesta

P: Antifeedant nature of the quinone primin and its quinol mi-

conidin from Miconia spp. Experientia 1984;40:1010–1011.

13. Cirilli R, Ferretti R, La Torre F, Borioni A, Fares V, Camalli M,

Faggi C, Rotili D, Mai A: Chiral HPLC separation and absolute

configuration of novel S-DABO derivatives. Chirality 2009;21:

604–612.

14. Lin K, Xu C, Zhou S, Liu W, Gan J: Enantiomeric separation of

imidazolinone herbicides using chiral high-performance liquid

chromatography. Chirality 2007;19:171–178.

15. Wagner HM, Bladt S, Zgainki EM: Plant Drug Analysis.

Springer, Berlin, 1984.

16. Conway WD: Countercurrent Chromatography—Apparatus,

Theory and Applications. VCH, New York, 1990.

17. Bilia AR, Ciampi L, Mendez J, Morelli I: Phytochemical in-

vestigations of Licania genus. Flavonoids from Licania pyrifolia.

Pharm Acta Helv 1996;71:199–204.

18. Ossipov V, Nurmi K, Loponen J, Prokopiev N, Haukioja E,

Pihlaja K: HPLC isolation and identification of flavonoids from

white birch Betula pubescens leaves. Biochem Syst Ecol 1995;23:

213–222.

19. Agrawal PK: Carbon-13 NMR of Flavonoids. Elsevier, Am-

sterdam, 1989.

20. Harborne JB: The Flavonoids: Advances in Research Since 1986.

Chapman and Hall, London, 1996.

21. Okuda T, Yoshida T, Hatano T, Koga T, Toh N, Kuriyama K:

Circular dichroism of hydrolysable tannins—I. Ellagitannins and

gallotannins. Tetrahedron Lett 1982;23:3937–3940.

22. Sadhu SK, Khatun A, Phattanawasina P, Ohtsuki T, Ishibashi M:

Lignan glycosides and flavonoids from Saraca asoca with anti-

oxidant activity. J Nat Med 2007;61:480–482.

23. Zhou X, Peng J, Fan G, Wu Y: Isolation and purification of fla-

vonoid glycosides from Trollius ledebouri using high-speed

counter-current chromatography by stepwise increasing the

flow-rate of the mobile phase. J Chromatogr A 2005;1092:216–

221.

24. Si W, Gong J, Tsao R, Kalab M, Yang R, Yin Y: Bioassay-

guided purification and identification of antimicrobial compo-

nents in Chinese green tea extract. J Chromatogr A 2006;1125:

204–210.

25. Dos Santos LC, Piacente S, De Ricardis F, Eletto AM, Pizza C,

Vilegas W: Xanthones and flavonoids from Leiothrix curvifolia

and Leiothrix flavescens. Phytochemistry 2001;56:853–856.

26. Sannomiya M, Rodrigues CM, Coelho RG, Dos Santos LC,

Hiruma-Lima CA, Brito ARMS, Vilegas W: Application of

preparative high-speed counter-current chromatography for the

separation of flavonoids from the leaves of Byrsonima crassa

Niedenzu (IK). J Chromatogr A 2004;1035:47–51.

27. Dos Santos LC, Silva MA, Rodrigues CM, Rodrigues J, Rinaldo

D, Sannomiya M, Vilegas W: [Fast preparative separation of

flavones from capitula of Eriocaulon ligulatum (Vell.) L.B.

Smith]. Rev Cienc Farm Basica Apl 2005;26:101–103.

28. Rinaldo D, Silva MA, Rodrigues CM, Calvo TM, Sannomiya M,

Dos Santos LC, Vilegas W, Kushima H, Hiruma-Lima CA, Brito

ARMS: [Preparative separation of flavonoids from the medicinal

plant Davilla elliptica St. Hill. by high-speed counter-current

chromatography]. Quim Nova 2006;29:947–949.

29. Conway WD, Petroski RJ: Modern Countercurrent Chromato-

graphy. American Chemical Society, Washington, DC, 1995.

30. Han X, Zhang T, Wei Y, Cao X, Ito Y: Separation of salidroside

from Rhodiola crenulata by high-speed counter-current chro-

matography. J Chromatogr A 2002;971:237–241.

31. Kodama S, Yamamoto A, Matsunaga A, Yanai H: Direct en-

antioseparation of catechin and epicatechin in tea drinks by 6-O-

a-d-glucosyl-b-cyclodextrin-modified micellar electrokinetic

chromatography. Electrophoresis 2004;25:2892–2898.

32. Slade D, Ferreira D, Marais JPJ: Circular dichroism, a powerful

tool for the assessment of absolute configuration of flavonoids.

Phytochemistry 2005;66:2177–2215.

33. Simões CMO, Schemkel EP, Gosmann G, Mello JCP, Mentz LA,

Petrovick PR: Farmacognosia: da Planta ao Medicament. Ed.

UFRGS, Porto Alegre=UFSC, Florianópolis, Brazil, 2001.

34. Yoshida T, Ito H, Hipólito IJ: Pentameric ellagitannin oligomers

in melastomataceous plants—chemotaxonomic significance.

Phytochemistry 2005;66:1972–1983.

35. Isaza JH, Ito H, Yoshida T: A flavonol glycoside-lignan ester and

accompanying acylated glucosides from Monochaetum multi-

florum. Phytochemistry 2001;58:321–327.

36. Dong L, Ni W, Dong J, Li J, Chen C, Liu H: A new neolignan

glycoside from the leaves of Acer truncatem. Molecules

2006;11:1009–1014.

37. Klyne W, Stevenson R, Swan RJ: Optical rotatory dispersion. 28.

Absolute configuration of otobain and derivatives. J Chem Soc C

1966;9:893–896.

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http://dx.doi.org/10.1016/j.phytol.2013.02.008