Analysis of rhizoma polygoni cuspidati by HPLC and HPLC-ESI/MS

Article (PDF Available)inPhytochemical Analysis 18(5):387-92 · September 2007with95 Reads
DOI: 10.1002/pca.993 · Source: PubMed
  • 1st Tao yi
    33.22 · Hong Kong Baptist University
  • 27.04 · Beijing University of Posts and Telecommunications
Abstract
An HPLC method with photodiode array detection (PAD) and ESI/MS detection was developed for the qualitative and quantitative analysis of the major chemical constituents of the dried rhizome of Polygonum cuspidatum Sieb. et Zucc. (Rhizoma Polygoni Cuspidati; Chinese name Hu-Zhang). Based on the chromatographic separation on an Altima C(18) column using 0.5% aqueous acetic acid and acetonitrile as the mobile phase, nine compounds, including stilbenes, stilbene glucosides, anthraquinones and anthraquinone glucosides, were identified by online ESI/MS analysis and seven were quantified by HPLC-PAD. A full validation of the method including sensitivity, linearity, repeatability and recovery was conducted. Linear calibration was achieved over the concentration range 1-200 mg/L with R(2) > 0.999, whilst the limits of detection ranged from 0.51 to 1.57 ng. Repeatability was evaluated by intra- and inter-day assays and the RSD value was within 1.79%. Recoveries of the quantified compounds were within the range 96.0-100.1% with RSD values of less than 2.2%. Five samples of Rhizoma Polygoni Cuspidati from different regions were analysed using the developed method. The major constituents piceid, resveratrol, emodin-8-O-beta-D-glucoside and emodin were selected to provide an index for the quality assessment of the herbal drug.
ANALYSIS OF THE DRIED RHIZOME OF POLYGONUM CUSPIDATUM 387
Copyright © 2007 John Wiley & Sons, Ltd. Phytochem. Anal. 18: 387–392 (2007)
DOI: 10.1002.pca
Phytochemical Analysis
Phytochem. Anal. 18: 387–392 (2007)
Published online 2 May 2007 in Wiley InterScience (www.interscience.wiley.com) DOI: 10.1002/pca.993
Analysis of Rhizoma Polygoni Cuspidati by HPLC and
HPLC-ESI/MS
TAO YI,
1
HAO ZHANG
1
and ZONGWEI CAI
2
*
1
West China School of Pharmacy, Sichuan University, No. 17, Section 3, Ren-Min-Nan-Lu Road, Chengdu, Sichuan 610041, People’s Republic of China
2
Department of Chemistry, Hong Kong Baptist University, Kowloon Tong, Kowloon, Hong Kong SAR, People’s Republic of China
Received 1 December 2005; Revised 17 January 2006; Accepted 12 January 2007
Abstract: An HPLC method with photodiode array detection (PAD) and ESI/MS detection was developed for the qualitative and
quantitative analysis of the major chemical constituents of the dried rhizome of Polygonum cuspidatum Sieb. et Zucc. (Rhizoma
Polygoni Cuspidati; Chinese name Hu-Zhang). Based on the chromatographic separation on an Altima C
18
column using 0.5%
aqueous acetic acid and acetonitrile as the mobile phase, nine compounds, including stilbenes, stilbene glucosides, anth-
raquinones and anthraquinone glucosides, were identified by online ESI/MS analysis and seven were quantified by HPLC-PAD.
A full validation of the method including sensitivity, linearity, repeatability and recovery was conducted. Linear calibration was
achieved over the concentration range 1–200 mg/L with R
2
> 0.999, whilst the limits of detection ranged from 0.51 to 1.57 ng.
Repeatability was evaluated by intra- and inter-day assays and the RSD value was within 1.79%. Recoveries of the quantified
compounds were within the range 96.0–100.1% with RSD values of less than 2.2%. Five samples of Rhizoma Polygoni Cuspidati
from different regions were analysed using the developed method. The major constituents piceid, resveratrol, emodin-8-O-
β
-D-
glucoside and emodin were selected to provide an index for the quality assessment of the herbal drug. Copyright © 2007 John
Wiley & Sons, Ltd.
Keywords: HPLC-UV; HPLC-ESI-MS; quality assessment; Polygonum cuspidatum: Rhizoma Polygoni Cuspidati.
INTRODUCTION
Rhizoma Polygoni Cuspidati (Hu-Zhang in Chinese
and Ko-Jo-Kon in Japanese) is the dried rhizome of
Polygonum cuspidatum Sieb. et Zucc. (Polygonaceae).
The drug has been widely used in Chinese and Japa-
nese folk medicine for the treatment of atherosclerosis,
hypertension, cough, suppurative dermatitis and gon-
orrhoea (Nonomura et al., 1963; State Pharmacopoeia
Commission of the People’s Republic of China, 2000).
Various classes of constituents have been isolated
from the drug, including anthraquinones, stilbenes,
flavonoids and phenols (Fig. 1; Xiao et al., 2002; Liu
et al., 2003; Hegde et al., 2004). The stilbenes have
been shown to be effective in the treatment of athero-
sclerosis whilst the anthraquinones possess antiviral
activities (Koyama et al., 2001; Hao and He, 2004).
Anthraquinones are the hydrolysis products of anth-
raquinone glucosides in an acid environment, whilst
stilbene glucosides are often converted to stilbenes
prior to sample analysis. Determination of anthraquin-
ones and stilbenes in Rhizoma Polygoni Cuspidati has
been carried out using scanning TLC (Zhao, 2005),
HPLC (Jiang et al., 2003) and capillary electrophoresis
(CE; Brandolini et al., 2002; Koyama et al., 2003).
However, the accurate and simultaneous qualitative
and quantitative analysis of these different groups of
compounds in a single chromatographic run has not
been previously reported. The current quality assess-
ment standard is merely based on the content of a
few marker compounds, namely, resveratrol-3-O-
β
-D-
glucoside (piceid) and/or resveratrol that have been
identified as bioactive compounds in the drug (Vastano
et al., 2000; Hao and He, 2004). However, it is widely
accepted that multiple constituents are responsible for
the therapeutic effect of herbs. Therefore, the current
quality assessment based on the contents of just one
or two marker compounds cannot accurately reflect the
quality of Rhizoma Polygoni Cuspidati (Normile, 2003;
Chan, 2005).
HPLC-UV and HPLC-MS provide online spectral
information for each peak in a chromatogram. The
techniques have been used to study peak purity by
evaluating the chromatograms and their associated MS
and are thus highly applicable to the routine quality
control of herbs. In the present study, an HPLC-PAD-
MS method for the qualitative and quantitative ana-
lysis of the major constituents in Rhizoma Polygoni
Cuspidati has been developed and fully validated.
According to the obtained results, four major constitu-
ents of the drug, namely, piceid, resveratrol, emodin-8-
O-
β
-D-glucoside and emodin, are suggested as the index
for quality assessment of this herbal drug.
Phytochemical
Analysis
* Correspondence to: Zongwei Cai, Department of Chemistry, Hong
Kong Baptist University, Kowloon Tong, Kowloon, Hong Kong SAR,
People’s Republic of China.
E-mail: zwcai@hkbu.edu.hk
388 T. YI ET AL.
Copyright © 2007 John Wiley & Sons, Ltd. Phytochem. Anal. 18: 387–392 (2007)
DOI: 10.1002.pca
Figure 1 Chemical structures of the constituents identified in Rhizoma Polygoni Cuspidati (the peak numbering refers to the
chromatograms shown in Figs 2 and 3).
EXPERIMENTAL
Materials.
HPLC-grade acetonitrile and acetic acid (used
in HPLC analysis), and analytical grade methanol (used
for sample extraction) were from Lab-scan, Bangkok,
Thailand. Deionized water used for HPLC analysis
was purified using a Milli-Q water purification system
(Millipore, Milford, MA, USA).
The standard reference compounds piceid, aloe-
emodin, emodin and physicoin were purchased from
the National Institute for the Control of Pharmaceutical
and Biological Products (Beijing, China). Resveratrol
was purchased from Sigma-Aldrich (St. Louis, MO,
USA). The standard reference compounds emodin-8-O-
β
-D-glucoside and physicoin-8-O-
β
-D-glucoside were
isolated from fresh rhizome of P. cuspidatum in our
laboratory using silica gel column chromatography,
preparative TLC and semi-preparative HPLC. Details
of their separation and structural elucidation will be
reported elsewhere.
Five samples of Rhizoma Polygoni Cuspidati were
collected from the Jiangsu, Hunan, Sichuan, Shanxi
and Gansu provinces of China. The identities of these
herbs were confirmed from morphological characteris-
tics, and microscopic and physical–chemical analysis
according to the Chinese Pharmacopoeia (State Phar-
macopoeia Commission of the People’s Republic of
China, 2000).
Sample preparation.
A representative portion of each
drug sample was ground to a powder and passed
through a 20 mesh (0.9 mm) sieve. An accurately
ANALYSIS OF THE DRIED RHIZOME OF POLYGONUM CUSPIDATUM 389
Copyright © 2007 John Wiley & Sons, Ltd. Phytochem. Anal. 18: 387–392 (2007)
DOI: 10.1002.pca
compounds. Repeatability was evaluated by intra-
and inter-day (n = 3) assays. Recoveries of all of the
quantified constituents were determined at different
concentration levels using a mixture of standards
containing 50, 100 and 200% of the quantified levels
of constituents in the samples. Replicated sample
analyses were conducted for the recovery studies. Five
drug samples, acquired in various regions of China,
were quantified using the described method.
RESULTS AND DISCUSSION
Detection wavelength for HPLC-UV analysis
By comparing the HPLC chromatograms of extracts
of Rhizoma Polygoni Cuspidati acquired at different
wavelengths within the range 210–500 nm, and the
corresponding UV absorption maxima for each stand-
ard compound, it was established that 320 nm was
the maximum absorption wavelength for piceid and
resveratrol, whilst 290 nm was optimal for anthra-
quinone glucosides as well as for anthraquinones.
Since there were no significant differences in the UV
absorbances of piceid and resveratrol at 290 and
320 nm, a wavelength of 290 nm was selected for
retrieving the HPLC profiles of the constituents. A
typical chromatogram obtained from the separation of
the major constituents of the drug is shown in Fig. 2.
HPLC–MS analysis of drug samples
The combined HPLC-UV and HPLC-MS technique can
provide online spectral information for each peak in
a chromatogram. In many cases, direct identification
of the peak is possible based on a comparison with
weighed sample (ca. 0.2 g) of powder was extracted
with 15 mL methanol by means of sonication at room
temperature for 30 min. The procedure was repeated
three times, the extracts combined in a 50 mL volu-
metric flask, and the whole made up to volume with
methanol. All of the samples were filtered through an
Alltech (Beerfield, IL, USA) syringe filter (0.2 µm) prior
to HPLC analysis. An aliquot of 10 µL solution was
injected for HPLC-PAD and HPLC-ESI/MS analysis.
HPLC–PAD analysis.
An Agilent (Palo Alto, CA, USA)
1100 series HPLC-PAD system, consisting of a vacuum
degasser, a binary pump, an autosampler, a thermo-
stated column compartment and a photodiode array
detector, was used for the quantitative analysis and for
UV spectra acquisition. The UV detector was set at
290 nm. For chromatographic analyses, an Altima
(Deerfield, IL, USA) C
18
column (250 × 4.6 mm i.d.;
5 µm) with a compatible guard column was used. The
mobile phase consisted of 0.5% acetic acid in water
(A) and acetonitrile (B) using a gradient program of
10–40% B in 0–35 min and 40–100% B in 35 60 min.
The mobile phase flow rate was 1 mL/min and the
column temperature was set at 30°C.
HPLC–ESI/MS analysis.
An Applied Biosystems (Foster
City, CA, USA) PE-SCIEX API 365 LC-MS system with
electrospray ionization source was used for mass
spectrometric determination. The ESI–MS spectra were
acquired in both the positive and negative ion modes.
The conditions of MS analysis were as follows: drying
gas (air) flow rate, 7 L/min; gas temperature, 300°C;
scan range, 50–500 amu; orifice voltage, 26 V; focusing
voltage, 170 V; and electrospray voltage, 5000 V.
Validation of quantitative method.
Calibration curves
were established for each of the standard reference
Figure 2 HPLC chromatogram of an extract of Rhizoma Polygoni Cuspidati with detection at 290 nm. Key to peak identity as in
Fig. 1. (For chromatographic protocol see Experimental section.)
390 T. YI ET AL.
Copyright © 2007 John Wiley & Sons, Ltd. Phytochem. Anal. 18: 387–392 (2007)
DOI: 10.1002.pca
Figure 3 TIC chromatogram of an extract of Rhizoma Polygoni Cuspidati obtained by HPLC-ESI/MS analysis in the positive ion
mode. Key to peak identity as in Fig. 1. (For chromatographic protocol see Experimental section.)
peaks were tentatively identified as torachrysone-8-
O-
β
-D-glucoside (peak 5) and chrysophanol-8-O-
β
-D-
glucoside (peak 7) by comparison of their m/z values
and UV spectra with data reported in the literature
(Xiao et al., 2002; Liu et al., 2003). The results obtained
(Table 1) indicated that the main types of constituents
in the drug were stilbene glucosides, stilbenes, anth-
raquinone glucosides and anthraquinones.
In the case of unknown HPLC peak 1, the ESI/MS
spectrum of the associated component exhibited three
ions at m/z 229 (relative intensity 63), 391 (100) and
413 (71). This indicates that the compound probably
has a molecular weight of 390, giving logical ion peaks
at m/z 229 for [M + H glu]
+
and m/z 413 for [M + Na]
+
.
For unknown peak 4, the positive ion spectrum exhib-
ited the same three peaks at m/z 229 (100), 391 (57)
and 413 (34), indicating that peaks 1 and 4 might cor-
respond to piceid isomers. Further characterisation of
these two compounds is in hand.
published data or standard compounds, and this has
been shown to be a powerful tool for the rapid identific-
ation of the constituents in plant extracts and herbal
products (Yi et al., 2005).
In the present study, the mass spectrometric condi-
tions were optimized in both the positive and negative
ion modes. The positive ion mode was, however, found
to be more sensitive. Most of constituents exhibited
quasi-molecular ions [M + H glu]
+
, [M + H]
+
, [M + Na]
+
and [M + K]
+
in the positive ion mode. Figure 3 shows
a total ion chromatogram obtained from the HPLC-
ESI/MS analysis of an extract of Rhizoma Polygoni
Cuspidati.
Based on the obtained m/z values, UV spectra and
comparison with standard compounds, seven peaks
were unambiguously identified as piceid (peak 2),
resveratrol (peak 3), emodin-8-O-
β
-D-glucoside (peak 6),
physcion-8-O-
β
-D-glucoside (peak 8), aloe-emodin (peak
9), emodin (peak 10) and physcion (peak 11). Two other
Table 1 Identification of components of Rhizoma Polygoni Cuspidati by HPLC-PAD and ESI-MS
Retention time [M + H – glu]
+
[M + H]
+
[M + Na]
+
[M + K]
+
Peak (min) Identification (m/z)(m/z)(m/z)(m/z)
λ
max
(nm)
1 15.229 Unknown compound 229 391 413 214, 304
2 18.634 Piceid 229 391 413 216, 320
3 22.613 Resveratrol 229 251 267 216, 318
4 24.702 Unknown compound 229 391 413 264, 404
5 28.047 Torachrysone-8-O-
β
-D-glucoside 247 409 431 447 216, 320
6 30.812 Emodin-8-
β
-D-glucoside 271 433 455 471 282, 424
7 33.544 Chrysophanol-8-O-
β
-D-glucoside 255 417 439 282, 424
8 35.188 Physcion-8-
β
-D-glucoside 285 447 469 485 282, 420
9 42.921 Aloe-emodin 271 293 290, 418
10 49.141 Emodin 271 293 309 288, 438
11 55.270 Physcion 285 307 286, 436
ANALYSIS OF THE DRIED RHIZOME OF POLYGONUM CUSPIDATUM 391
Copyright © 2007 John Wiley & Sons, Ltd. Phytochem. Anal. 18: 387–392 (2007)
DOI: 10.1002.pca
Table 2 Calibration curves and LOD of seven constituents of Rhizoma Polygoni Cuspidati determined at 290 nm
Peak Compound Slope (A) Intercept (B) R
2
LOD (ng)
2 Piceid 33.621 2.4765 0.9998 1.27
4 Resveratrol 22.437 1.5732 0.9998 0.78
6 Emodin-8-
β
-D-glucoside 16.888 +1.8257 0.9997 1.22
8 Physcion-8-
β
-D-glucoside 13.486 +1.5425 0.9998 1.57
9 Aloe-emodin 23.362 1.3440 0.9990 0.51
10 Emodin 37.127 1.6480 0.9995 0.63
11 Physcion 19.582 0.9729 0.9990 0.82
Table 3 Repeatability of the described method for the determination of constituents of Rhizoma Polygoni Cuspidati
First day Third day Fifth day
Calculated content calculated content calculated content Inter-day
Compound (mg/g)
a
(mg/g) (mg/g) RSD (%)
Piceid 1.75 ± 0.38 1.74 ± 0.39 1.75 ± 0.42 0.33
Resveratrol 0.378 ± 0.53 0.376 ± 0.61 0.374 ± 0.64 0.53
Emodin-8-
β
-D-glucoside 3.69 ± 0.21 3.71 ± 0.28 3.70 ± 0.17 0.27
Physcion-8-
β
-D-glucoside 0.299 ± 0.54 0.303 ± 0.69 0.298 ± 0.44 0.88
Aloe-emodin 0.032 ± 1.24 0.032 ± 0.95 0.033 ± 1.23 1.79
Emodin 1.05 ± 0.25 1.03 ± 0.31 1.05 ± 0.27 1.11
Physcion 0.180 ± 0.84 0.176 ± 0.74 0.181 ± 0.89 1.48
a
The values shown are mean ± RSD (n = 3).
Validation of quantitative method
Validation of the developed method was carried out
and the results are listed in Tables 24. For all of
the quantified constituents, good linear calibrations
were achieved over the concentration range 1–200 mg/L
with R
2
> 0.999. The RSD values were found to be
within the ranges 0.21–1.24% for intra-day assays and
0.27–1.79% for inter-day assays. At a signal-to-noise
ratio of 3:1, the limits of detection (LOD) of the quanti-
fied constituents were determined to be within the
range 0.51–1.57 ng. The average recoveries of all of
the quantified constituents were within the range
96.0–100.1% with RSD 2.2%.
Five Rhizoma Polygoni Cuspidati samples acquired
from different regions of China were determined using
the developed method. The results (Table 5) demon-
strate that the contents of the quantified constitu-
ents varied as a result of the differences in cultivation
conditions.
The results obtained from the HPLC-PAD-MS ana-
lysis show that the major chemical constituents in
Rhizoma Polygoni Cuspidati consist of two stilbene
compounds and two anthraquinone compounds, in-
cluding piceid, resveratrol, emodin-8-
β
-D-glucoside
and emodin, which is in good agreement with reports
that the drug has multiple efficacies as an anti-
atherosclerotic and an antiviral agent (Wang and
Table 4 Recovery study of seven constituents of Rhizoma Polygoni Cuspidati
Recovery of seven constituents (%)
a
Spike level Emodin-8-
β
- Physcion-8-
β
-
(%) Piceid Resveratrol D-glucoside D-glucoside Aloe-emodin Emodin Physcion
50 98.5 ± 0.9 97.4 ± 0.9 100.2 ± 1.2 98.4 ± 1.5 94.3 ± 2.1 96.4 ± 1.2 96.5 ± 1.3
100 102.2 ± 1.3 98.5 ± 1.3 98.6 ± 0.9 95.7 ± 1.3 95.2 ± 2.0 98.9 ± 1.0 98.4 ± 1.1
200 99.6 ± 1.1 99.8 ± 1.2 98.0 ± 0.8 97.4 ± 1.2 98.4 ± 1.8 97.3 ± 0.9 100.3 ± 1.0
Mean 100.1 ± 1.9 98.6 ± 1.2 98.9 ± 1.1 97.2 ± 1.4 96.0 ± 2.2 97.5 ± 1.3 98.4 ± 1.9
a
The values shown are mean ± RSD (n = 3).
392 T. YI ET AL.
Copyright © 2007 John Wiley & Sons, Ltd. Phytochem. Anal. 18: 387–392 (2007)
DOI: 10.1002.pca
Table 5 Contents of seven constituents in five different samples of Rhizoma Polygoni Cuspidati
Content (mg/g)
a
Compound RPC-1 RPC-2 RPC-3 RPC-4 RPC-5
Piceid 1.75 ± 0.38 2.87 ± 0.43 5.03 ± 0.36 4.82 ± 0.34 4.71 ± 0.37
Resveratrol 0.378 ± 0.53 0.715 ± 0.59 1.12 ± 0.45 1.15 ± 0.55 1.05 ± 0.53
Emodin-8-
β
-D-glucoside 3.69 ± 0.21 6.53 ± 0.17 9.49 ± 0.15 9.60 ± 0.16 9.04 ± 0.18
Physcion-8-
β
-D-glucoside 0.299 ± 0.54 0.642 ± 0.47 0.727 ± 0.48 0.833 ± 0.41 0.854 ± 0.46
Aloe-emodin 0.032 ± 1.24 0.106 ± 0.76 0.076 ± 0.92 0.100 ± 0.70 0.109 ± 0.82
Emodin 1.05 ± 0.25 2.50 ± 0.22 1.38 ± 0.27 1.82 ± 0.19 2.06 ± 0.17
Physcion 0.180 ± 0.84 0.456 ± 0.77 0.204 ± 1.08 0.286 ± 0.87 0.337 ± 0.77
a
The values shown are mean ± SD (n = 3): drug samples RPC-1 to RPC-5 were collected from the Jiangsu, Hunan, Sichuan,
Shanxi, Gansu provinces of China, respectively.
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Therefore, these four chemical compounds may be
selected as the index for quality assessment of this
herbal drug.
The present study describes a facile method for the
identification and quantification of the major chemical
components in Rhizoma Polygoni Cuspidati. The ana-
lytical procedure is precise and reproducible, and is
thus suitable for the analysis of a large number of
samples. Based on the obtained results, it is suggested
that two stilbene compounds and two anthraquinone
compounds, namely piceid, resveratrol, emodin-8-
β
-D-
glucoside and emodin, be used as the index for quality
assessment of this herbal drug.
Acknowledgements
The authors would like to thank Dr Zhonghui Li
(Department of Chemistry, Hong Kong Baptist Univer-
sity) for technical support on the NMR and EI-MS
experiments.
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2277.
    • "glikozid B) azonosítottak (2. ábra) (Beerling et al. 1994, Yi et al. 2007, Qian et al. 2008, Kovárová et al. 2010, Kovárová et al. 2011, Shen et al. 2011, Peng et al. 2013, Chen et al. 2013, Frantík et al. 2013, Békési-Kallenberger et al. 2013 ). A levelek és a szár fl avonoidokat, fl avonolokat és antociánokat tartalmaznak (Beerling et al. 1994, Murai et al. 2015). "
    [Show abstract] [Hide abstract] ABSTRACT: A Fallopia sectio Reynoutria (óriáskeserűfű) fajok kelet-ázsiai eredetűek, de napjainkban a világ számos pontján előfordulnak. Európába a XIX. században dísznövényként kerültek be egyes fajok. Fő hatóanyagaik között cserzőanyagok, fenolkarbonsavak, flavonoidok, stilbének (pl. rezveratrol) és antrakinonok (p. emodin) említhetők, amelyek számos élettani hatással és gyógyászati jelentőséggel is rendelkeznek. A tradicionális kínai és japán medicina egyes fajok rizómáját hepatitis, magas vérnyomás, bőrsérülések és vérzés ellen alkalmazza. Napjainkban számos vizsgálat igazolja a növények baktérium- és gombaölő, antioxidáns, citotoxikus, neuroprotektív, koleszterinszint- és gyulladáscsökkentő hatását, továbbá rákterápiában való alkalmazásukra is rendelkezésre állnak adatok. Egyes taxonok nehézfémkötő szereppel és allelopátiás hatással is jellemezhetők, míg mások takarmányként való termesztésére folynak kísérletek. Kiemelendő a fajok energianövényként való alkalmazása is, amely magas fűtőértékükkel áll összefüggésben. Ezek az eredmények további ökológiai, fitokémiai és hatástani vizsgálatok lehetőségét vetítik előre, amelyek a nemzetség számos potenciális felhasználási területére irányítják a figyelmet. Összefoglalónkban a japán (F. japonica), a szahalini (F. sachalinensis) és Európában keletkezett hibridjük, a cseh óriáskeserűfű (F. ×bohemica) történeti, etnobotanikai, morfológiai, hisztológiai, fő fitokémiai, népgyógyászati, valamint gyógyászati jelentőségét tekintjük át. — ABSTRACT: Fallopia species originate from Asia, but nowadays they are spreading in many areas of the world. In Europe, taxa have been planted in the 19th century as ornamental plants. Among the main phytochemical compounds, tannins, phenolic acids, flavonoids, stibenes (e.g. resveratrol) and anthraquinones (e.g. emodin) can be highlighted, which have several physiological and therapeutical effects, as well. In the traditional Chinese and Japanese medicine, the rhizome of some species is used for hepatitis, high blood pressure, skin injuries and bleeding. Recently, several studies have been carried out on the antibacterial, antifungal, antioxidant, anti-inflammatory, and neuroprotective effect of the plants. In addition, preliminary pharmacological data are available on their effect against high cholesterol level and for some cancer types, as well. Some species have metal-binding property and allelopathic effect, while other taxa are studied for their agricultural production to use as fodder. The possible role of Fallopia species as energy plants is based on their high heating value. These results suggest further ecological, phytochemical and pharmacological studies in the future, which draw the attention for the various potential uses of these plants. This work summarizes the botanical (incl. morphological, histological, phytochemical and medicinal), as well as the historical and ethnobotanical features of Fallopia japonica, F. sachalinensis and their hybrid F. ×bohemica.
    Full-text · Article · Jan 2016 · Journal of Chromatography A
    • "The identification of peaks 2, 5, 10, and 11 was further confirmed by comparing their retention time values with those of the standards. Emodin, polydatin, and resveratrol can be identified unambiguously by comparing the MS data (m/z 269, 389, and 227) and the retention times (44.8, 8.8, and 15.7 min), respectively , with those of the literature and authentic compounds [28]. These compounds amount to 29.6, 22.4, and 2.55% of total extract content, respectively (Table 3). "
    [Show abstract] [Hide abstract] ABSTRACT: Resistance of cancer cells to chemotherapy is controlled by the decrease of intracellular drug accumulation, increase of detoxification, and diminished propensity of cancer cells to undergo apoptosis. ATP-binding cassette (ABC) membrane transporters with intracellular metabolic enzymes contribute to the complex and unresolved phenomenon of multidrug resistance (MDR). Natural products as alternative medicine have great potential to discover new MDR inhibitors with diverse modes of action. In this study, we characterized several extracts of traditional Chinese medicine (TCM) plants ( = 16) for their interaction with ABC transporters, cytochrome P3A4 (CYP3A4), and glutathione-S-transferase (GST) activities and their cytotoxic effect on different cancer cell lines. Fallopia japonica (FJ) (Polygonaceae) shows potent inhibitory effect on CYP3A4 P-glycoprotein activity about 1.8-fold when compared to verapamil as positive control. FJ shows significant inhibitory effect (39.81%) compared with the known inhibitor ketoconazole and 100 μg/mL inhibited GST activity to 14 μmol/min/mL. FJ shows moderate cytotoxicity in human Caco-2, HepG-2, and HeLa cell lines; IC50 values were 630.98, 198.80, and 317.37 µg/mL, respectively. LC-ESI-MS were used to identify and quantify the most abundant compounds, emodin, polydatin, and resveratrol, in the most active extract of FJ. Here, we present the prospect of using Fallopia japonica as natural products to modulate the function of ABC drug transporters. We are conducting future study to evaluate the ability of the major active secondary metabolites of Fallopia japonica to modulate MDR and their impact in case of failure of chemotherapy.
    Full-text · Article · Aug 2015
    • "The chromatograms revealed significant enhancement of 10 peaks (P1–P10) compared with the control containing denatured mitochondria (P > 20%, shown inTable 2), demonstrating specific binding to active mitochondria. By analyzing the UV, MS and MS n data provided by LC/MS (Table 2) and comparison with previously reported data363738 and standards, the P1–P10 peaks were confirmed as piceid (P1), polygonimitin B (P2), epigallocatechin gallate (P3), 3,5,4 -trihydroxystilben-3-O- (6 -galloyl)-glucoside (P4), emodin-1-O-glucoside (P5), resveratrol (P6), torachryson-8-O-glucoside (P7), emodin-8-O-glucoside (P8), emodin-8-O-(6 -malonyl)-glucoside (P9) and emodin (P10), respectively, which are classified as five anthraquinone ( "
    [Show abstract] [Hide abstract] ABSTRACT: Mitochondria are an important intracellular pharmacological target because damage to this organelle results in a variety of human disorders and because mitochondria are involved in complex processes such as energy generation, apoptosis and lipid metabolism. To expedite the search for natural bioactive compounds targeting mitochondria, we initially developed an efficient mitochondria-based screening method by combining centrifugal ultrafiltration (CU) with liquid chromatography/mass spectrometry (LC/MS), which is called screening method for mitochondria-targeted bioactive constituents (SM-MBC) and is compatible with the search of mitochondria-targeted compounds from complex matrixes such as herbal medicines extracts. Functionally active, structurally intact and pure mitochondria were obtained from rat myocardium using an optimized protocol for mitochondrial isolation comprising organelle release followed by differential and Nycodenz density gradient centrifugation. After evaluating the reliability of the method using thiabendazole (TZ), rotenone (RN), amiodarone (AR) and trimetazidine (TD) as positive controls, this method was successfully applied to screen bioactive constituents from extracts of Polygoni Cuspidati Rhizoma et Radix (PCRR) and Scutellariae Radix (SR). Nineteen active compounds were detected and identified by LC/MS, of which 17 were new mitochondria-targeted compounds. The activity of 9 of the 19 hit compounds was confirmed by in vitro pharmacological trials. These results demonstrate that SM-MBC can be used for the efficient screening of mitochondria-targeted constituents in complex preparations used to treat mitochondrial disorders, such as PCRR and SR. The results may be meaningful for an in-depth understanding of drug mechanism of action and drug discovery from medicinal herbs. Copyright © 2015 Elsevier B.V. All rights reserved.
    Full-text · Article · Aug 2015
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