全 文 ： 170 Journal of Chinese Pharmaceutical Sciences http://www.jcps.ac.cn
Simultaneous determination of eight flavonoids in Polygonum aviculare L.
by RP-HPLC-UV
Qingrong Fu, Shujuan Liu, Hong Wang, Shizhong Chen*
Department of Natural Medicines, School of Pharmaceutical Sciences, Peking University Health Science Center, Beijing 100191, China
Abstract: A reliable high performance liquid chromatography method was developed for the quality evaluation of Polygonum
aviculare L. Eight marker flavonoids were identified and simultaneously quantified, which included myricitrin, hyperoside,
galuteolin, avicularin, quercitrin, quercetin, luteolin, and kaempferol. The analysis was performed on an Inertsil ODS-4 column
(4.6 mm×150 mm, 5 µm) with gradient elution. The mobile phases were 0.5% aqueous phosphoric acid and acetonitrile. The
detection wavelength was 360 nm. The eight marker flavonoids were separated well with good linearity (r2>0.9991), precision,
stability and repeatability. The recovery rate was 95.58%–102.65%. Cluster analysis was employed to analyze 28 batches of
samples. The result indicated that this method provides an efficient way to perform quality control as well as a scientific rationale
for the Geo-authentication of Polygonum aviculare L.
Keywords: Polygonum aviculare L.; Flavonoids; Quantitative analyses; Cluster analysis
CLC number: R284.1 Document code: A Article ID: 1003–1057(2014)3–170–07
Received: 2013-07-24; Revised: 2013-10-13; Accepted: 2013-10-28.
Foundation item: Study of Nature of Geo-authentic Crude Drug
(“973” State Key Project, Grant No. 2006CB504700).
*Corresponding author. Tel.: 86-10-82802723; Fax: 86-10-82802723;
E-mail: chenshizhong66@163.com
http://dx.doi.org/10.5246/jcps.2014.03.022
1. Introduction
Polygonum aviculare L. (P. aviculare) is widely distributed
in Asia, Africa, Latin America and Middle Eastern
countries and used as folkloric medicine. In China, it has
been used as a diuretic and detoxifying medicine for
the treatment of atopic eczema, strangurtia, itching,
gonorrhea and several inflammatory diseases[1]. Previous
phytochemical and pharmacological studies have demon-
strated that flavonoids[2,3] are the main bioactive compounds of
P. aviculare, which possess a wide array of pharmacological
activities such as antidiabetic[4], anti-gingivitis[5], anti-
microbial[6], anti-inflammatory[7], antioxidant[8,9] and
recovery of acetaminophen-induced nephrotoxicity[10].
A series of methods have been developed for the deter-
mination of active compounds in P. aviculare, including
UV-vis spectrophotometry[11,12] and high performance
liquid chromatography (HPLC)[13–15]. However, these
methods exhibited low resolution, low sensitivity and
detection of fewer analytes (less than five compounds).
Moreover, they cannot be used for the comprehensive
evaluation of the quality of P. aviculare.
It is well known that the therapeutic effects of traditional
Chinese medicines (TCMs) are usually attributed to
multiple bioactive compounds[16,17]. And herbs produced
in different areas vary in efficacy due to the intrinsic nature
and content of active ingredients. Therefore, simultaneous
determination of multiple components in P. aviculare is
required for the quality control of dosage during clinical
studies. The chromatographic fingerprint was also regarded as
a useful method to control the quality of herbal medicines
and their derivatives because it emphasizes on the systemic
characterization of compositions in samples and focuses
on the identification and assessment of the stability of
the components.
Until now, there are few reports[18,19] on the quantifica-
tion of multiple bioactive compounds and the fingerprints
of P. aviculare. In this study, a reliable and comprehensive
quality assessment method was developed to generate
HPLC fingerprints for the simultaneous determination of
eight major flavonoids in 28 batches of P. aviculare samples
collected from different regions. The quantification of
eight marker compounds was used for the quality control
of P. aviculare. Moreover, cluster analysis was performed
to guide the Geo-authentication study. This study provides
an effective method for the comprehensive quality assessment
of P. aviculare.
2. Experimental
2.1. Materials, chemicals and reagents
Twenty-eight bathes of P. aviculare were collected from
various locations in different provinces of China. Sample
S8 was used for the method development. The materials
were authenticated by Prof. Hong Wang (Pharmaceutical
Sciences, Peking University Health Science Center, Beijing,
171 Fu, Q.R. et al. / J. Chin. Pharm. Sci. 2014, 23 (3), 170–176
China) and voucher specimens were deposited at the School
of Pharmaceutical Sciences, Peking University, China.
Standard compounds of myricitrin, hyperoside, galuteolin,
avicularin, quercitrin, quercetin, luteolin, and kaempferol
(Fig. 1) were purchased from the National Institute for
the Control of Pharmaceutical and Biological Products
(Beijing, China).
HPLC-grade acetonitrile (Fisher, USA) was used for
HPLC analysis. The deionized water was prepared with
a Millipore water purification system (Millipore, Milford,
MA, USA) and was filtered with 0.22 µm membranes.
HPLC-grade phosphoric acid and analytical-grade
absolute ethanol for sample preparation were purchased
from Beijing Chemical Plant.
2.2. Preparation of standard solutions
Mixed stock solution containing myricitrin, hyperoside,
galuteolin, avicularin, quercitrin, quercetin, luteolin, and
kaempferol was prepared in 60% ethanol. Working
standard solutions were prepared by diluting the mixed
standard solution with 60% ethanol to give different
concentrations for the construction of calibration curves.
The solutions were filtered through a 0.22 µm membrane
prior to injection.
2.3. Sample preparation
The accurately weighed powder (0.5 g, 40-mesh) was
extracted with 25 mL 60% ethanol in an ultrasonic bath
for 30 min (250 W, 40 kHz). After the samples were
cooled to room temperature, the 60% ethanol lost during
extraction was made up, and the extracts were filtered
with a 0.22 µm membrane filter prior to HPLC analysis.
2.4. HPLC-DAD analysis
Generation of HPLC fingerprints and quantitative
analyses were performed on an Shimadzu LC-10A liquid
chromatography system (Shimadzu International Trading
Co., Ltd., Japan), equipped with a double pump, a degasser,
an autosampler, a DAD detector, and a thermostated
column compartment. The separations were carried out
on an Inertsil ODS-4 column (4.6 mm×150 mm, 5 µm);
Shimadzu-GL sciences (Shanghai laboratory supplies
Co., Ltd.) at 45 °C. The mobile phases consisted of
0.5% aqueous phosphoric acid (A) and acetonitrile (B)
with the gradient condition as follows: 10%–18% B (v/v)
at 0–10 min, 18%–19% B at 10–25 min, 19%–23% B at
25–60 min, 23%–45% B at 60–75 min. The flow rate
was 1.0 mL/min and the injection volume was 10 µL.
Detection wavelength was set at 360 nm using DAD.
2.5. Method validation
2.5.1. Calibration curves, limits of detection and
quantification
Calibration curves were generated by plotting the
concentrations of the mixed standard solutions (X, mg/mL)
versus the peak areas (Y). The limit of detection (LOD)
and the limit of quantification (LOQ) were determined at
the signal-to-noise ratio of about 3 and 10, respectively.
2.5.2. Precision, repeatability, stability and recovery
The intra-and inter-day precisions were investigated by
determining a mixed standard solution in six replicates on
the same day and by duplicating the experiments on three
consecutive days. The repeatability was determined by
analyzing six replicates with the above established
method. Stability of sample solution was analyzed at 0, 2,
4, 8, 12, 24 and 48 h within 2 days at room temperature,
respectively. Also, a recovery study was used to evaluate
the accuracy of the method. Recoveries were determined
by spiking accurately known amounts of the eight-analyte
solution to approximately 0.5 g of the P. aviculari powder
prior to the extraction.
3. Results and discussion
3.1. Optimization of sample preparation
In order to obtain satisfactory extraction efficiency,
extraction method (ultrasonication and heat-reflux),
extraction solvent (40%, 60% and 100% ethanol) and
extraction time (15, 30, 45 and 60 min) were optimized.
The optimal sample preparation was found to be the
extraction of 0.5 g sample powder with 25 mL of 60%
ethanol in an ultrasonic water bath for 30 min.
3.2. Optimization of chromatographic conditions
HPLC conditions including detection wavelength and
O
HO
OH
OH
OH
OR
O
HO
OR
OH
OH
O
HO
OH
OH
OH
OH
OR
O
HO
OH
OH
OH
R = H Quercetin R = H Luteolin
R = Rha Quercitrin R = Glu Galuteolin
R = Gal Hyperoside
R = Ara Avicularin
R = Rha Myricitrin Kaempferol
Figure 1. Chemical structures of the investigated compounds.
172 Fu, Q.R. et al. / J. Chin. Pharm. Sci. 2014, 23 (3), 170–176
mobile phase were investigated for the optimization of
chromatographic separations. The wavelength chosen must
cover more characteristic peaks and show high absorption
and resolution. As a result, 360 nm was selected as the
optimal detection wavelength by a DAD full wavelength
scan (200–400 nm) and data reported in the literature[19].
Various mixtures of 0.5% aqueous phosphoric acid and
acetonitrile were screened in order to obtain a reliable
chromatogram with most peaks at acceptable resolutions
and to obtain baseline separation of each marker compound
in a relatively short analytical time. Better separation was
also found when the column temperature was maintained
at 45 °C. The optimal HPLC conditions used in this study
are shown in Section 2.4.
3.3. HPLC fingerprints of P. aviculare
Using the optimized HPLC method, eight peaks (1, 2,
3, 4, 5, 6, 7 and 8) in the HPLC fingerprint profile were
structurally identified as myricitrin, hyperoside, galuteolin,
avicularin, quercitrin, quercetin, luteolin, and kaempferol,
respectively, by comparing their retention times, UV
absorption with those of reference standards and in some
cases data reported in the literature. The separation of
eight flavonoids and a typical HPLC chromatogram are
shown in Figure 2.
3.4. Validation of the method
3.4.1. Calibration curves, limits of detection and
quantification
Calibration curves were generated by plotting the
concentrations of the mixed standard solutions (X, mg/mL)
with the peak areas (Y). The limit of detection (LOD) and
the limit of quantification (LOQ) were determined at
a signal-to-noise ratio of about 3 and 10, respectively.
The calculated results are summarized in Table 1. All the
analytes showed good linearity (r2>0.9991) in a wide concen-
tration range. The LOD and LOQ of the eight compounds
were 0.04–2.39 and 1.40–7.98 µg/mL, respectively.
3.4.2. Precision, repeatability, stability and recovery
The intra-and inter-day precisions were investigated by
determining a mixed standard solution in six replicates
during a single day and by duplicating the experiments on
three consecutive days. The repeatability was determined
by analyzing six replicates with the above established
method. Stability of the sample solution was analyzed at 0,
2, 4, 8, 12, 24 and 48 h within 2 days at room temperature.
Moreover, a recovery test was used to evaluate the accuracy
of the method. The results are shown in Table 2. Recoveries
were determined by spiking accurately known amounts
(low, medium and high) of the eight-analyte solution to
Figure 2. The typical HPLC chromatographic profile of P. aviculare (A), mixed standards (B), and comparison of S8, S12, S15 and S28.
The peaks marked with 1, 2, 3, 4, 5, 6, 7 and 8 are myricitrin, hyperoside, galuteolin, avicularin, quercitrin, quercetin, luteolin, and
kaempferol, respectively. The separation condition was described in Section 2.2.
(A) 3.5
3.0
2.5
2.0
1.5
1.0
0.5
0.0
mAU (×10)
360 nm, 4 nm (1.0)
1
2
3
4
5
6 7 8
0 10 20 30 40 50 60 70
t (min)
(B) 2.5
2.0
1.5
1.0
0.5
0.0
mAU (×10)
360 nm, 4 nm (1.0)
0 10 20 30 40 50 60 70
t (min)
(C) 5.0
4.0
3.0
2.0
1.0
0.0
uV (×104)
0 10 20 30 40 50 60 70
t (min)
S8
S12
S15
S28
173 Fu, Q.R. et al. / J. Chin. Pharm. Sci. 2014, 23 (3), 170–176
approximately 0.5 g of the P. aviculare powder prior to
the extraction (Table 3). The results indicated that the
intra and inter-day precision, repeatability, stability and
recovery RSD values of the eight compounds were all less
than 2.81%. The recovery of the analytes was in the range
of 95.58%–102.65%. These validation results indicated that
the established method for the quantitative determination of
these eight flavonoids was acceptable.
3.5. Quantitative determination of eight marker
compounds in P. aviculare
The established method was subsequently applied to the
simultaneous determination of the eight marker compounds
in P. aviculare from different regions. Each sample was
analyzed in triplicate to determine the mean content (mg/g)
and the results are presented in Table 4.
Table 1. Linear regression analysis, LOD and LOQ of eight flavonoids
Table 2. Precision, repeatability and stability of the eight flavonoids
Table 3. Recoveries of eight flavonoids in P. aviculare
Analytes tR (min) Calibration equation r2 Linear range (µg/mL) LOD (µg/mL) LOQ (µg/mL)
Myricitrin 18.03 y = 7.2879×107x + 7.0200×104 0.9999 34.2–307.8 1.95 6.51
Hyperoside 18.61 y = 5.1846×107x – 2.8941×104 0.9992 5.16–46.44 1.37 4.55
Galuteolin 19.28 y = 5.7947×107x – 4.4911×104 0.9991 4.32–38.88 1.25 3.46
Avicularin 23.59 y = 4.5441×107x + 3.0031×103 0.9999 23.20–208.80 1.92 6.39
Quercitrin 25.67 y = 9.5545×107x + 3.5292×103 0.9992 18.48–166.32 2.39 7.98
Quercetin 48.74 y = 2.3523×107x + 2.1414×103 0.9995 5.80–52.20 1.24 4.13
Luteolin 50.53 y = 1.4474×107x + 5.5345×104 0.9999 1.48–13.32 0.04 1.40
Kaempferol 70.22 y = 2.6018×107x + 3.5466×104 0.9997 4.68–42.12 0.65 2.18
Analytes
Precision (RSD, %)
Repeatability (RSD%, n = 6) Stability (RSD%, n = 7)
Intra-day (n = 6) Inter-day (n = 3)
Myricitrin 0.07 0.58 0.62 0.38
Hyperoside 0. 13 1.32 1.29 0.92
Galuteolin 0. 60 2.53 2.74 0.32
Avicularin 0.04 0.77 0.78 0.85
Quercitrin 0.04 0.75 0.845 0.63
Quercetin 1.84 1.34 1.82 2.44
Luteolin 1.52 2.76 2.60 2.70
Kaempferol 0. 70 2.58 2.26 2.08
Analytes Original (µg) Spiked (µg) Observed (µg)
Recovery (%) RSD (%)
x Average x Average
Myricitrin 1315
1048 2349 98.66
101.53
1.43
1.78 1310 2661 102.75 1.89
1572 2937 103.18 2.02
Hyperoside 85
76 159 97.36
99.93
2.33
2.71 95 179 98.94 2.87
114 203 103.50 2.93
Galuteolin 40
37.6 78 101.06
100.58
2.03
2.10 47 86 97.87 1.99
56.4 98 102.83 2.28
Avicularin 160
123.2 282 100.65
101.20
0.98
1.07 154 312 100.65 1.18
184.8 350 102.27 1.05
Quercitrin 675
532 1207 100.00
100.86
2.37
2.81 665 1343 100.45 2.70
798 1490 102.13 3.35
Quercetin 65
56.8 123 102.11
102.65
1.01
1.03 71 137 101.40 1.02
85.2 154 104.46 1.05
Luteolin 20
17.6 37 96.59
95.58
2.99
2.78 22 41 95.45 2.46
26.4 45 94.70 2.89
Kaempferol 60
48.8 110 102.45
102.63
3.17
2.77 61 122 101.63 2.44
73.2 136 103.82 2.69
174 Fu, Q.R. et al. / J. Chin. Pharm. Sci. 2014, 23 (3), 170–176
Sample No. Locations
Conents (mg/g crude drugs, n = 3)
Myricitrin Hyperoside Galuteolin Avicularin Quercitrin Quercetin Luteolin Kaempferol
S1 Fuzhou, Fujian 2.63 0.17 0.08 0.32 1.35 0.13 0.04 0.12
S2 Xi’an, Shaanxi 0.51 0.03 0.12 0.31 0.42 0.09 0.07 0.03
S3 Sanming, Fujian 1.44 0.08 0.06 0.49 0.75 0.08 0.04 0.07
S4 Baoji, Shaanxi 1.03 0.05 0.06 0.33 0.54 0.09 0.04 0.03
S5 Xianyang, Shaanxi 1.06 0.04 0.12 0.35 0.68 0.09 0.04 0.11
S6 Zhengzhou, Henan 1.00 0.07 0.05 0.35 0.53 0.09 0.03 0.05
S7 Xishui, Hubei 1.94 0.20 0.12 0.85 1.03 0.04 0.03 0.05
S8 Bozhou, Anhui 1.53 0.11 0.10 0.69 0.83 0.09 0.02 0.08
S9 Ganzhou, Jiangxi 1.22 0.08 0.11 0.77 0.71 0.13 0.02 0.09
S10 Hangzhou, Zhejiang 1.86 0.14 0.32 0.76 0.85 0.08 0.03 0.06
S11 Tonghua, Jilin 1.51 0.10 0.06 0.58 0.74 0.07 0.02 0.06
S12 Nanyang, Henan 1.15 0.06 0.09 0.43 0.66 0.12 0.03 0.08
S13 Xinyang, Henan 1.22 0.08 0.04 0.46 0.65 0.09 0.04 0.08
S14 Linfen, Shanxi 0.93 0.02 0.03 0.26 0.50 0.05 0.03 0.06
S15 Datong, Shanxi 0.73 0.02 0.08 0.19 0.42 0.07 0.03 0.03
S16 Beijing 0.85 0.04 0.04 0.48 0.50 0.09 0.02 0.06
S17 Wuchang, Heilongjiang 2.01 0.20 0.06 0.38 0.86 0.11 0.05 0.11
S18 Chengdu, Sichuan 2.02 0.07 0.06 0.47 1.06 0.13 0.06 0.10
S19 Zhoukou, Henan 0.60 0.01 0.02 0.17 0.38 0.08 0.03 0.04
S20 Zhengzhou, Henan 0.72 0.05 0.04 0.30 0.32 0.07 0.03 0.03
S21 Beijing 0.57 0.19 0.16 0.15 0.38 0.08 0.02 0.04
S22 Yancheng, Jiangsu 1.18 0.08 0.08 0.69 0.58 0.16 0.02 0.10
S23 Hefei, Anhui 1.37 0.13 0.07 0.73 0.68 0.09 0.03 0.06
S24 Yiyang, Hunan 1.50 0.16 0.09 0.74 0.78 0.06 0.03 0.05
S25 Changzhou, Jiangsu 1.47 0.16 0.10 0.73 0.79 0.06 0.02 0.05
S26 Ningbo, Zhejiang 1.41 0.11 0.08 0.75 0.78 0.16 0.03 0.09
S27 Qingdao, Shandong 1.62 0.10 0.06 0.72 0.74 0.12 0.02 0.09
S28 Guangzhou, Guangdong 1.67 0.22 0.13 1.01 0.91 0.12 0.02 0.06
Table 4. Sample information and quantitative determination of eight flavonoids in 28 P. aviculare samples and their calculated similarity values
Significant variations in the amount of the 8 compounds
among samples were observed. Myricitrin (0.51–2.63 mg/g),
avicularin (0.15–1.01 mg/g) and quercitrin (0.32–1.35 mg/g)
were the most dominant constituents in each sample,
which is consistent with a previous report[18]. However,
the contents of quercetin (0.04–0.16 mg/g) and luteolin
(0.02–0.07 mg/g) were much lower. A possible explanation
is that almost all other determinations of quercetin and
luteolin were performed after acid hydrolysis of the
corresponding glycosides[13]. In addition, concentrations
of the compounds varied significantly among the samples.
The hyperoside content of S28 was 22-fold higher than
that of S19. The galuteolin content of S10 was 16-fold
higher than that of S19. This result suggests that the
source greatly affected the contents of ingredients, which
may lead to different efficacy. Therefore, appropriate
cultivation areas should be selected to ensure the safety
and effectiveness of P. aviculare products used for
therapeutic purposes. Obvious variations could also be
found in other constituents. Results from this study agree
with a previous study that chromatographic fingerprinting
combined with quantitative analysis of marker compounds
is a better tool for the quality evaluation of traditional
Chinese medicine[20].
In summary, eight flavonoids have been identified and
simultaneously quantified in P. aviculare, which provides
a basic starting point for enriching and improving the
quality standard of P. aviculare. For the other compounds
identified on HPLC fingerprint, the quantification cannot
be done mainly because the chemical structures of these
compounds have not been identified yet. We are separating
and preparing these compounds, and planning to conduct
further research in the future.
3.6. Cluster analysis of P. aviculare samples
Data from 28 batches of P. aviculare samples collected
from various regions in China were subjected to cluster
analysis using SPSS 19.0 software. All samples were
classified on the basis of the contents of the eight marked
compounds. Figure 3 shows a dendrogram of the hierar-
chical cluster analysis of the 28 tested samples using the
ward’s method. The horizontal axis indicates the Euclidean
distances (which represent differences between samples),
175 Fu, Q.R. et al. / J. Chin. Pharm. Sci. 2014, 23 (3), 170–176
whereas the vertical axis represents the sample numbers.
As shown by the dendrogram, the samples were obviously
divided into two clusters when the Euclidean distance
was 25. Cluster I mainly includes S2, S4, S5, S14, S15,
S6, S12, S13, S19, S20, S16 and S21 collected in
Shaanxi, Shanxi, Henan Provinces and Beijing City,
which were located in the Loess Plateau and the Northern
China. Cluster II contain the rest of the samples, which
were collected from the eastern or coastal region of the
country. Loess Plateau and the North China experience
northern temperate continental monsoon climate, of which,
winter and spring are cold, dry and sandy due to the air
from the polar region, while summer and autumn are hot
and rainy because of the Northwest Pacific subtropical
high pressure and the Indian Ocean low pressure. The
annual precipitations of these places are less than 800 mm,
especially for that of the Loess Plateau (about 466 mm).
However, P. aviculare are moisture loving plants and
a dry condition would have significantly impact on the
growth of the plants, like the accumulation of flavonoids.
Thus, P. aviculare from these areas exhibited lower
content of the eight marked compounds and were grouped
into one type. Samples of cluster II grow in the rainy
environment, with the annual rainfall of over than 800 mm,
which is beneficial for the accumulation of flavonoids.
Overall, the results of the cluster analysis provide a scientific
rationale for the study of Geo-authentication of P. aviculare.
Furthermore, we investigated the effect of the eight
marked flavonoids in P. aviculare on the hierarchical
cluster analysis using the method of principal components
analysis. The result showed that it is the contents of
myricitrin, hyperoside, galuteolin and avicularin that
basically determine the clusters, indicating that the
contents of the four flavonoids are the main factors that
influence the quality of P. aviculare. Comparison of
typical fingerprints of cluster I (S12 and S15) and cluster
II (S8 and S28) are shown in Figure 2.
4. Conclusion
Quality evaluation of traditional medicines is essentially
different from that of chemical drugs because of their
complexity and multiple-component nature. It has been
recognized that the therapeutic effects of traditional Chi-
nese medicine are due to the synergistic contribution of
multiple components, not just by the major constituents.
Thus, it is essential to establish a method that can determine
most of the chemical constituents for the quality control
of herbal medicines. This study was the first report on the
HPLC fingerprint and simultaneous quantification of eight
flavonoid compounds in P. aviculare from different regions.
Moreover, hierarchical cluster analysis and principal
component analysis provide an efficient and comprehensive
tool for the Geo-authentication study and quality evaluations
of P. aviculare. The method developed in this study can
be extended to the comprehensive quality assessment of
other traditional Chinese medicines.
Acknowledgements
The research was supported by a grant from the Study
of Nature of Geo-authentic Crude Drug (“973” State Key
Project) (Grant No. 2006CB504700).
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RP-HPLC-UV法同时测定萹蓄中8个黄酮类化合物的含量
付庆荣, 刘淑娟, 王弘, 陈世忠*
北京大学医学部 药学院 中药研究室, 北京 100191
摘要: 建立了反相高效液相色谱法同时测定扁蓄药材中8个黄酮 (杨梅苷, 金丝桃苷, 木犀草苷, 扁蓄苷, 槲皮苷, 槲皮素,
木犀草素和山奈酚)的含量。采用Inertsil ODS-4 色谱柱, 乙腈–0.5%磷酸水溶液梯度洗脱, 流速为1 mL/min, 柱温为35 ºC,
检测波长为360 nm。上述8个黄酮类成分达到基线分离, 标准曲线线性关系良好 (r2>0.9991), 日内和日间精密度、稳定性
和重复性实验均符合要求, 加样回收率为95.58%–102.65%。对28批不同产地的扁蓄样品进行了含量测定和聚类分析, 结果
表明该方法不仅可以对萹蓄药材进行质量控制, 对于萹蓄药材的道地性分析也提供了科学数据。
关键词: 萹蓄; 黄酮; 定量分析; 聚类分析