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  • 1 Kunming University of Science and Technology, Kunming 650500, Yunnan, China
  • 2 Kunming University of Science and Technology, Kunming 650500, Yunnan, China
  • 3 Kunming University of Science and Technology, Kunming 650093, China
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An effective, reliable, and sensitive reversed-phase high-performance liquid chromatography (RP-HPLC) with diode array detector (DAD) method was investigated for simultaneous determination of polydatin, isoquercitrin, resveratrol, and nicotiflorin in Tetrastigma hemsleyanum. The chromatographic separation of the four compounds was carried out on a Welchrom ODS column (4.6 mm × 250 mm, 5 μm) by gradient elution with phosphoric acid (H3PO4) aqueous solution (0.4%)–methanol as the mobile phase, at the temperature of 30 °C and a flow rate of 1.0 mL/min. The detection wavelength was set at 270 nm. Under optimum conditions, the baseline separation of these four compounds can be performed within 30 min. The developed method was validated in terms of detection limit, quantification limit, linearity, precision, and recovery tests. Eventually, the established HPLC–DAD method was successfully applied to the simultaneous determination of polydatin, isoquercitrin, resveratrol, and nicotiflorin in the extract of herb T. hemsleyanum.

Abstract

An effective, reliable, and sensitive reversed-phase high-performance liquid chromatography (RP-HPLC) with diode array detector (DAD) method was investigated for simultaneous determination of polydatin, isoquercitrin, resveratrol, and nicotiflorin in Tetrastigma hemsleyanum. The chromatographic separation of the four compounds was carried out on a Welchrom ODS column (4.6 mm × 250 mm, 5 μm) by gradient elution with phosphoric acid (H3PO4) aqueous solution (0.4%)–methanol as the mobile phase, at the temperature of 30 °C and a flow rate of 1.0 mL/min. The detection wavelength was set at 270 nm. Under optimum conditions, the baseline separation of these four compounds can be performed within 30 min. The developed method was validated in terms of detection limit, quantification limit, linearity, precision, and recovery tests. Eventually, the established HPLC–DAD method was successfully applied to the simultaneous determination of polydatin, isoquercitrin, resveratrol, and nicotiflorin in the extract of herb T. hemsleyanum.

Introduction

Tetrastigma hemsleyanum belongs to the grape family Vitaceae, known as “Sanyeqing” in China. T. hemsleyanum has been applied to the treatment of tumors and several other diseases, such as high fever, infantile febrile convulsion, neumonia, snake bite, and jaundice. Studies have examined the anticancer [15], liver protection, antioxidant [6], anti-inflammatory, analgesic, antipyretic activities [7], and chemical components of the root tubers [810]. Moreover, some studies researched chemical components and biological activities of the leaves [11, 12]. Petroleum ether fraction (PEF) is a major part that shows several biological activities, including anti-inflammatory [13], anti-hypoxicactivity [14], anti-nociceptiveactivity [15], anti-bacterial [16], and anti-tumor [17] activities in Chinese folk medicine.

T. hemsleyanum has been reported to contain a number of phytochemicals such as phenolic acids, flavonoids, flavanols, and phytosterols [1820]. Among these compounds, flavonoids have been considered to be the most significant bioactive constituents in T. hemsleyanum [20]. It has been clarified that flavonoids could display various bioactivities, such as anti-inflammatory function, antiviruses, and antiproliferative effect [21, 22], and may be responsible for the pharmacological activities of plants. Nonetheless, the full profiles of phytochemicals of T. hemsleyanum have not been reported before. Additionally, less study was focused on the systematic analysis of the chemical constituents. Consequently, identification and characterization of the chemical constituents in T. hemsleyanum could help to guarantee the safe and reliable use of T. hemsleyanum.

Different types of analytical methods have been exploited for the analysis of effective components in T. hemsleyanum, such as liquid chromatography coupled with ultraviolet detector (LC–UV) [23], gas chromatography coupled with mass spectrometry (GC–MS) [24], liquid chromatography coupled with triple-quadrupole tandem mass spectrometry (LC–MS/MS) [25], liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (LC–Q-TOF-MS), and liquid chromatography triple quadrupole tandem mass spectrometry (LC–QqQ-MS) [26]. High-performance liquid chromatography (HPLC) is a widely used method in terms of simplicity, rapidity, and high efficiency. Diode array detector (DAD) has many advantages, such as high selectivity, high sensitivity, and rapid analysis. The high-performance liquid chromatography (HPLC) with diode array detector (DAD) method has been successfully applied for the analysis of Chinese herbal medicines and biological samples because polydatin, isoquercitrin, resveratrol, and nicotiflorin possess benzene rings, which offer a strong ultraviolet absorption. To the best of our knowledge, the simultaneous determination of polydatin, isoquercitrin, resveratrol, and nicotiflorin in the roots, leaves, and fruits of T. hemsleyanum by HPLC–DAD has not been reported yet. The aim of this study was to develop a reliable and sensitive HPLC–DAD method for the simultaneous determination of polydatin, isoquercitrin, resveratrol, and nicotiflorin in T. hemsleyanum. Under optimized conditions, the method was validated and successfully applied to examine the contents of polydatin, isoquercitrin, resveratrol, and nicotiflorin in the roots, leaves, and fruits of T. hemsleyanum from Mile county (Yunnan, China). The results showed that the developed HPLC–DAD method could be a helpful tool for quality control of T. hemsleyanum. Structural formulae of polydatin, isoquercitrin, resveratrol, and nicotiflorin are presented in Figure 1.

Figure 1.
Figure 1.

Chemical structures of (A) polydatin, (B) isoquercitrin, (C) resveratrol, and (D) nicotiflorin

Citation: Acta Chromatographica Acta Chromatographica 30, 3; 10.1556/1326.2017.00231

Experimental

Instruments

An Ultimate 3000 HPLC system (Dionex, Sunnyvale, CA, USA) equipped with an Ultimate 3000 diode array detector (Dionex, Sunnyvale, CA, USA) was applied for HPLC analysis. Data were collected by Dionex Chromeleon software. The chromatographic separation was performed on a Welchrom ODS column (4.6 mm × 250 mm, 5 μm; Welch Materials, Shanghai, China).

Chemicals and reagents

Polydatin, isoquercitrin, resveratrol, and nicotiflorin were purchased from the National Institute for the Control of Pharmaceutical and Biological Products (Beijing, China). The roots, leaves, and fruits of T. hemsleyanum were collected from Mile County (Yunnan, China). The HPLC-grade methanol was purchased from Merck (Merck Co., Darmstadt, Germany). Deionized water was purified using a Milli-Q system from Millipore (Bedford, MA, USA). Other chemicals and solvents used in sample preparation were of analytical grade.

Preparation of standard solutions

Standard stock solutions of polydatin, isoquercitrin, resveratrol, and nicotiflorin were prepared by dissolving 8 mg of each analyte in 10 mL of methanol to achieve a concentration of 0.8 mg/mL and stored away from light at 4 °C. Working standard solutions were prepared by diluting each of the standard stock solutions with methanol to appropriate concentration range for the establishment of calibration curves. The final concentration range was 2–40 ng/mL for polydatin, 3–200 ng/mL for isoquercitrin, 2–100 ng/mL for resveratrol, and 3–100 ng/mL for nicotiflorin. Because polydatin, isoquercitrin, resveratrol, and nicotiflorin contain easily oxidized hydroxyl groups, the sample was kept away from the light. All solutions were filtered through 0.45 μm nylon membrane (Millipore, Milfored, MA, USA) before HPLC analysis.

Measurement of ultraviolet–visible properties

The ultraviolet–visible spectra of polydatin, isoquercitrin, resveratrol, and nicotiflorin were measured in methanol by a diode array detector. The standard solutions of these four compounds were diluted to appropriate concentrations to acquire good peak shapes.

Chromatographic conditions

Chromatographic analysis was performed on an Ultimate 3000 HPLC system (Dionex, Sunnyvale, CA, USA), consisting of a three-dimensional pump solvent management system, an on-line degasser, an autosampler, and an Ultimate 3000 diode array detector. Chromatography software of Dionex Chromeleon was used to collect and process the chromatographic data. A reversed-phase column (Welchrom ODS) was applied to separate some solutes. The mobile phase consisted of methanol (A) and 0.4% phosphoric acid aqueous solution (B) using a linear gradient program of 30–45% (A) in 0–16 min and 45–50% (A) in 16–30 min. The flow rate of the mobile phase was set at 1.0 mL/min, and the temperature was maintained at 30 °C. Detection wavelength was set at 270 nm, and the injection volume was 10 μL. All solutions were filtered before used.

Sample preparation

The samples of T. hemsleyanum were comminuted (100 mesh). Two grams of each sample was weighed accurately and added into a calibrated flask with 25 mL methanol. The mixture was subjected to ultrasonic bath at room temperature with methanol for 30 min and made up the loss of weight with methanol after cooling. Then, all solutions were filtered. Eventually, 10 μL of the extract solutions was injected into the HPLC–DAD system for analysis.

Results and discussion

Optimization of extracting conditions

Satisfactory extraction efficiency was obtained by comparing reflux extraction, soxhlet extraction, and ultrasonic extraction methods, according to the withdrawal rates of polydatin, isoquercitrin, resveratrol, and nicotiflorin; the ultrasonic extraction was chosen as the extracting method. Moreover, the pulverized samples were ultrasonic extracted for 10, 30, 50, and 60 min, respectively, and the maximum peak areas of polydatin, isoquercitrin, resveratrol, and nicotiflorin were acquired after 30 min of extraction. With longer extraction time, no effective improvements were obtained. The ultrasonic extraction temperature was set at room temperature because the high temperature destroyed the structures of the four compounds. In addition, the pulverized samples were ultrasonic extracted for 30 min with ethyl acetate, ethanol, methanol, and deionized water, respectively. By comparing the peak areas and peak heights of polydatin, isoquercitrin, resveratrol, and nicotiflorin in the extraction solution gained from the above four solvents, it was found that, upon extraction, the peaks of polydatin, isoquercitrin, resveratrol, and nicotiflorin were much higher in methanol than those in ethyl acetate, ethanol, and deionized water. As a result, the best extraction condition was established as follows: the samples were extracted for 30 min by ultrasonic extraction using methanol as the extraction solvent, and the extraction temperature was set at room temperature.

Ultraviolet–visible properties of polydatin, isoquercitrin, resveratrol, and nicotiflorin

In the present study, as shown in Figure 2, the detection wavelength was selected according to the maximum adsorption wavelengths of polydatin, isoquercitrin, resveratrol, and nicotiflorin at 307, 257, 306, and 266 nm, respectively, shown in UV spectra with three-dimensional chromatograms of diode array detector. It was found that, at the absorption wavelength of 270 nm, polydatin, isoquercitrin, resveratrol, and nicotiflorin could all present high peaks for simultaneous determination. Consequently, the absorption wavelength for the simultaneous detection of polydatin, isoquercitrin, resveratrol, and nicotiflorin was set at 270 nm to acquire the highest sensitivity.

Figure 2.
Figure 2.

Ultraviolet–visible spectra of (A) polydatin, (B) isoquercitrin, (C) resveratrol, and (D) nicotiflorin

Citation: Acta Chromatographica Acta Chromatographica 30, 3; 10.1556/1326.2017.00231

Optimization of HPLC conditions

In general, a suitable chromatographic column, mobile phase, and elution mode are critically important for good separation. In this study, an Ultimate 3000 HPLC system equipped with an Ultimate 3000 diode array detector was applied for separation and detection. Different columns packed with different materials, Kromasil 100-5C18 column (250 mm), Hypersil C18 column (250 mm), and Welchrom C18 column (250 mm) were employed, and it was investigated that the best separation effects were acquired by using the Welchrom C18 column (250 mm). The mobile phase consisting of water–methanol or water–acetonitrile could not completely separate polydatin, isoquercitrin, resveratrol, and nicotiflorin. Using methanol and 0.4% phosphoric acid aqueous solution as the mobile phase could improve the separation efficiency. Normally, the concentrations of phosphoric acid aqueous solution have effects on the charge conditions of the analytes, further influencing the retention time of the analytes on chromatographic columns. Accordingly, it is essential to optimize the concentration of phosphoric acid aqueous solution. The results exhibited that, when using a linear gradient program of 30–45% (methanol) in 0–16 min and 45–50% (methanol) in 16–30 min with a flow rate of 1.0 mL/min, the separation of polydatin, isoquercitrin, resveratrol, and nicotiflorin could gain good resolutions, good peak shapes, and reasonable testing time (within 30 min). The typical chromatograms of polydatin, isoquercitrin, resveratrol, and nicotiflorin are shown in Figure 3A.

Figure 3.
Figure 3.

Chromatograms of (A) standard sample, (B) root extract, (C) fruit extract, (D) leaf extract. Peaks: 1, polydatin; 2, isoquercitrin; 3, resveratrol; 4, nicotiflorin

Citation: Acta Chromatographica Acta Chromatographica 30, 3; 10.1556/1326.2017.00231

Calibration curves and the limit of detection

All calibration curves were plotted based on linear regression analysis of the integrated peak areas (Y) versus concentrations (X, ng/mL) of the four constituents in the standard solution at different concentrations. The regression equations, correlation coefficients, linear ranges, the limit of detection (LOD), and limit of quantification (LOQ) for the analysis of the four constituents are listed in Table 1. These four constituents showed good linearity in the range of 2–40 ng/mL for polydatin, 3–200 ng/mL for isoquercitrin, 2–100 ng/mL for resveratrol, and 3–100 ng/mL for nicotiflorin. The correlation coefficients were 0.99990, 0.99994, 0.99990, and 0.99990 for polydatin, isoquercitrin, resveratrol, and nicotiflorin, respectively, which guaranteed the good linearity of four compounds in the corresponding range. LODs (S/N = 3) for polydatin, isoquercitrin, resveratrol, and nicotiflorin were observed to be as low as 1.1, 1.3, 1.6, and 1.8 ng/mL, respectively. LOQs (S/N = 10) for polydatin, isoquercitrin, resveratrol, and nicotiflorin were 3.5, 4.1, 5.2, and 5.6 ng/mL, respectively.

Table 1.

Analytical performance data for of four analyes by HPLC–DAD

CompoundRetention times (min)Calibration range (ng/mL)Regression equationR2LOD (ng/mL)LOQ (ng/mL)
Polydatin14.1232-40Y = 125.379X0.999901.13.5
Isoquercitrin21.7703-200Y = 98.9764X0.999941.34.1
Resveratrol22.4072-100Y = 108.942X0.999901.65.2.
Nicotiflorin27.6173-100Y = 184.3342X0.999901.85.6

Precision, stability, repeatability and accuracy

The relative standard deviation (RSD) was taken as a measure of precision, accuracy, and stability. Intra-day and inter-day precisions were researched by six replicated injections (n = 6) of the mixture standard solutions (10 ng/mL of polydatin, 10 ng/mL of isoquercitrin, 20 ng/mL of resveratrol, and 20 ng/mL of nicotiflorin) in 1 day and one injection each day for six consecutive days. RSDs (intra- and inter-day) of the retention time and peak area were less than 2% and 4%, respectively. The low RSD values guaranteed the good precision (Table 2). The same real sample was examined within 24 h for the stability assay; the RSD values of the retention time and peak area were both lower than 2%. As a result, the solution was stable.

Table 2.

Precision and accuracy for the detection of four components by HPLC–DAD

ComponentsRSD of retention time (%)RSD of peak area (%)
Intra-dayInter-dayIntra-dayInter-day
Polydatin0.030.281.213.51
Isoquercitrin0.061.291.053.43
Resveratrol0.120.601.373.64
Nicotiflorin0.201.131.453.95

The injection of six different samples which were obtained through the same sample preparation procedure was investigated for the repeatability. The results were shown in Table 3, the RSD values of the repeatability were lower than 0.1% for the retention time, and the RSD values were lower than 2% for the peak area.

Table 3.

Repeatability for the detection of four components by HPLC–DAD

ComponentsRSD of retention time (%)RSD of peak area (%)
Polydatin0.061.81
Isoquercitrin0.081.65
Resveratrol0.051.46
Nicotiflorin0.091.98

With the purpose of valuing the accuracy of the developed HPLC–DAD method, the recovery test was performed with the roots of T. hemsleyanum. The recovery of the investigated components ranged from 98.1% to 103.8%, and their RSD values were all less than 2.3% (Table 4). The recovery experiments manifested the developed method has fine accuracy for the measurement of these components. Meanwhile, the results revealed that the proposed HPLC–DAD method can be applied in the quality control of the traditional Chinese medicine T. hemsleyanum.

Table 4.

Precision and accuracy for the detection of four components in Tetrastigma hemsleyanum

CompoundSample weight (g)Contents (μg)Added (μg)Found (μg)Recovery (%)RSD (%)
Polydatin2.028135.27315.269810.442898.12.24
2.029905.27775.269810.6845102.6
2.196625.71125.269810.9968100.3
Isoquercitrin2.0281312.574412.593925.6469103.82.16
2.0299012.585412.593925.2926100.9
2.1966213.619012.593926.149999.5
Resveratrol2.028137.09857.125114.131098.71.94
2.029907.10467.125114.3295101.4
2.196627.68827.125114.9914102.5
Nicotiflorin2.0281369.970572.0362142.9432101.31.13
2.0299070.031672.0362142.5721100.7
2.1966275.783472.0362147.171399.1

Application

In order to examine the application for practical analysis, the developed method was used for the determination of polydatin, isoquercitrin, resveratrol, and nicotiflorin in T. hemsleyanum samples extract from Mile County (Yunnan, China). The roots, leaves, and fruits of T. hemsleyanum extracts were shown as representatives in Figure 3. The results were summarized in Table 5, indicating that the concentrations of analytes differ among the roots, leaves, and fruits, in general; the contents of the four measured components in the leaf samples were the highest. Because the growth of the roots, leaves, and fruits of T. hemsleyanum can be affected by the conditions of soil, water, and climate, the result was reasonable. The result that the contents of every component differ among the roots, leaves, and fruits of T. hemsleyanum in China revealed that the production parts of the medicine should be taken into consideration when utilizing T. hemsleyanum as a Chinese herb to treat specific diseases. For instance, with respect to the functions of anti-inflammatory and antiproliferative, the leaves of T. hemsleyanum are preferred.

Table 5.

Content comparisons of four components in the different parts of Tetrastigma hemsleyanum

No.PartPolydatinIsoquercitrinResveratrolNicotiflorin
Contents (μg/g)RSD (%)Contents (μg/g)RSD (%)Contents (μg/g)RSD (%)Contents (μg/g)RSD (%)
1Root2.60.96.20.33.50.434.50.4
2Root3.50.85.90.73.10.520.50.5
3Root2.91.24.70.51.21.311.61.2
4Root3.80.74.20.54.71.244.10.6
5Root5.41.13.80.92.50.430.50.8
6Root4.31.35.41.33.60.841.20.8
7Root3.30.53.11.22.00.624.50.4
8Fruit5.01.42.60.71.40.90.51.2
9Fruit4.31.03.81.31.81.00.40.9
10Fruit5.71.72.41.02.50.91.11.5
11Fruit3.80.62.90.82.00.80.81.0
12Fruit4.50.85.40.53.81.50.51.4
13Fruit5.51.24.20.92.40.80.70.7
14Fruit3.00.82.81.11.41.00.30.8
15Leaf4.80.98235.10.5352.71.1334.50.6
16Leaf5.80.74624.00.8452.51.3423.71.3
17Leaf4.20.85645.80.6234.11.2420.60.8
18Leaf7.41.13565.30.9235.70.9283.41.0
19Leaf8.80.87523.50.8275.00.7537.90.8
20Leaf10.81.34523.91.3305.80.5235.30.5
21Leaf7.80.86584.80.6287.11.1421.90.7

Conclusions

In this work, an HPLC–DAD method was established and validated for the simultaneous determination of polydatin, isoquercitrin, resveratrol, and nicotiflorin. The HPLC–DAD method of T. hemsleyanum developed in this study is precise, efficient, practical, and reliable and, therefore, could be used in the quality control of T. hemsleyanum. Additionally, this method is convenient for proper clinical utilization and further pharmacological investigation of this herb.

Acknowledgment

This work was supported by Yunnan Province Project Education Fund of China (No. 2015Y085) and the Analysis Test Fund of Kunming University of Science and Technology, Yunnan Province, China (No. 2016T20070029).

References

  • 1.

    Feng Z. Q. ; Hao W. R.; Lin X. Y.; Fan D. P.; Zhou J. H. Onco Targets Ther. 2014, 7, 947956.

  • 2.

    Feng Z. Q. ; Ni K. F.; He Y.; Ding Z. S.; Zhu F.; Wu L. C.; Shen M. H. J. Chin. Clin. Pharmacol. Therapeut. 2006, 11, 669672.

  • 3.

    Liu Y. Y. ; Xia H. J. Hunan Norm. Univ. 2010, 7, 2224.

  • 4.

    Wang X. ; Zeng J.; Zhou H. Anti-tumor Phar. 2012, 2, 347350.

  • 5.

    Zhong L. R. ; Chen X.; Wei K. M. Asian Paci. J. Cancer Prevent. 2013, 14, 59835987.

  • 6.

    Sun Y. ; Li H. Y.; Hu J. N.; Li J.; Fan Y. W.; Liu X. R.; Deng Z. Y. J. Agric. Food. Chem. 2013, 61, 1050710515.

  • 7.

    Huang Z. ; Mao Q. Q.; Wei J. P. J. Chin. New Drugs 2005, 14, 861864.

  • 8.

    Li Y. ; Lu W.; Yu Z. Chin. Trad. Herb. Drugs 2003, 34, 982983.

  • 9.

    Liu D. ; Ju J. H.; Lin G.; Xu X. D.; Yang J. S.; Tu G. Z. Acta Bota. Sinica. 2001, 44, 227229.

  • 10.

    Liu D. ; Yang J. Chin. J. Chin. Mater. Medi. 1999, 24, 611612.

  • 11.

    Hossain M. A. ; Shah M. D.; Gnanaraj C.; Iqbal M. Asian Paci. J. Trop. Medi. 2011, 4, 717721.

  • 12.

    Shao Q. S. ; Deng Y. M.; Shen H. J.; Fang H. L.; Zhao X. F. Int. J. Biol. Macromol. 2011, 49, 958962.

  • 13.

    Guo D. J. ; Xu L. J.; Cao X. W.; Guo Y. Q.; Ye Y.; Chan C. O.; Mok D. K. W.; Yu Z. L.; Chen S. B. J. Ethnopharmacol. 2011, 138, 717722.

  • 14.

    Ma H. P. ; Fan P. C.; Jing L. L.; Yao J.; He X. R.; Yang Y.; Chen K. M.; Jia Z. P. J. Ethnopharmacol. 2011, 137, 15101515.

  • 15.

    Chen Y. F. ; Li N.; Jiao Y. L.; Wei P.; Zhang Q. Y.; Rahman K.; Zheng H. C.; Qin L. P. Phytomedicine 2008, 15, 427436.

  • 16.

    Zhang Y. Q. ; Xu J.; Yin Z. Q.; Jia R. Y.; Lu Y.; Yang F.; Du Y. H.; Zou P.; Lv C.; Hu T. X.; Liu S. L.; Shu G.; Yi G. Fitoterapia 2010, 81, 747750.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 17.

    Cao X. L. ; Xu J.; Bai G.; Zhang H.; Liu Y.; Xiang J. F.; Tang Y. L. J. Chromatogr. B 2013, 929, 610.

  • 18.

    Hu Y. Q. ; Cheng L.; Pu J. B.; Liang W. Q.; Zheng J. X. Chin. J. Trad. Medi. Sci. Tech. 2013, 20, 4647.

  • 19.

    Li Y. Q. ; Lu W. C.; Yu Z. G. Chin. Trad. Herb. Drugs 2003, 34, 982983.

  • 20.

    Liu D. Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China, 2000.

  • 21.

    Coppin J. P. ; Xu Y.; Chen H.; Pan M. H.; Ho C. T.; Juliani R.; Simon J. E.; Wu Q. J. Func. Foods 2013, 5, 18921899.

  • 22.

    Xu C. J. ; Ding G. Q.; Fu J. Y.; Meng J.; Zhang R. H.; Lou X. M. Biomed. Environ. Sci. 2008, 21, 325331.

  • 23.

    Liu Y. ; Qian L. H. Anhui Agri. Sci. Bull. 2015, 21, 2628.

  • 24.

    Liu L. J. ; Song W. F. Clin. Med. Engin. 2011, 18, 18571858.

  • 25.

    Xu W. ; Fu Z. Q.; Lin J.; Huang X. C.; Yu H. M.; Huang Z. H.; Fan S. M. Acta Pharmaceut. Sinica. 2014, 49, 17111717.

  • 26.

    Xu W. ; Fu Z.Q.; Lin J.; Huang X. C.; Chen D.; Yu H. M.; Huang Z. H.; Fan S. M. Chin. J. Chin. Mater. Med. 2014, 39, 43654372.

If the inline PDF is not rendering correctly, you can download the PDF file here.

  • 1.

    Feng Z. Q. ; Hao W. R.; Lin X. Y.; Fan D. P.; Zhou J. H. Onco Targets Ther. 2014, 7, 947956.

  • 2.

    Feng Z. Q. ; Ni K. F.; He Y.; Ding Z. S.; Zhu F.; Wu L. C.; Shen M. H. J. Chin. Clin. Pharmacol. Therapeut. 2006, 11, 669672.

  • 3.

    Liu Y. Y. ; Xia H. J. Hunan Norm. Univ. 2010, 7, 2224.

  • 4.

    Wang X. ; Zeng J.; Zhou H. Anti-tumor Phar. 2012, 2, 347350.

  • 5.

    Zhong L. R. ; Chen X.; Wei K. M. Asian Paci. J. Cancer Prevent. 2013, 14, 59835987.

  • 6.

    Sun Y. ; Li H. Y.; Hu J. N.; Li J.; Fan Y. W.; Liu X. R.; Deng Z. Y. J. Agric. Food. Chem. 2013, 61, 1050710515.

  • 7.

    Huang Z. ; Mao Q. Q.; Wei J. P. J. Chin. New Drugs 2005, 14, 861864.

  • 8.

    Li Y. ; Lu W.; Yu Z. Chin. Trad. Herb. Drugs 2003, 34, 982983.

  • 9.

    Liu D. ; Ju J. H.; Lin G.; Xu X. D.; Yang J. S.; Tu G. Z. Acta Bota. Sinica. 2001, 44, 227229.

  • 10.

    Liu D. ; Yang J. Chin. J. Chin. Mater. Medi. 1999, 24, 611612.

  • 11.

    Hossain M. A. ; Shah M. D.; Gnanaraj C.; Iqbal M. Asian Paci. J. Trop. Medi. 2011, 4, 717721.

  • 12.

    Shao Q. S. ; Deng Y. M.; Shen H. J.; Fang H. L.; Zhao X. F. Int. J. Biol. Macromol. 2011, 49, 958962.

  • 13.

    Guo D. J. ; Xu L. J.; Cao X. W.; Guo Y. Q.; Ye Y.; Chan C. O.; Mok D. K. W.; Yu Z. L.; Chen S. B. J. Ethnopharmacol. 2011, 138, 717722.

  • 14.

    Ma H. P. ; Fan P. C.; Jing L. L.; Yao J.; He X. R.; Yang Y.; Chen K. M.; Jia Z. P. J. Ethnopharmacol. 2011, 137, 15101515.

  • 15.

    Chen Y. F. ; Li N.; Jiao Y. L.; Wei P.; Zhang Q. Y.; Rahman K.; Zheng H. C.; Qin L. P. Phytomedicine 2008, 15, 427436.

  • 16.

    Zhang Y. Q. ; Xu J.; Yin Z. Q.; Jia R. Y.; Lu Y.; Yang F.; Du Y. H.; Zou P.; Lv C.; Hu T. X.; Liu S. L.; Shu G.; Yi G. Fitoterapia 2010, 81, 747750.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 17.

    Cao X. L. ; Xu J.; Bai G.; Zhang H.; Liu Y.; Xiang J. F.; Tang Y. L. J. Chromatogr. B 2013, 929, 610.

  • 18.

    Hu Y. Q. ; Cheng L.; Pu J. B.; Liang W. Q.; Zheng J. X. Chin. J. Trad. Medi. Sci. Tech. 2013, 20, 4647.

  • 19.

    Li Y. Q. ; Lu W. C.; Yu Z. G. Chin. Trad. Herb. Drugs 2003, 34, 982983.

  • 20.

    Liu D. Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China, 2000.

  • 21.

    Coppin J. P. ; Xu Y.; Chen H.; Pan M. H.; Ho C. T.; Juliani R.; Simon J. E.; Wu Q. J. Func. Foods 2013, 5, 18921899.

  • 22.

    Xu C. J. ; Ding G. Q.; Fu J. Y.; Meng J.; Zhang R. H.; Lou X. M. Biomed. Environ. Sci. 2008, 21, 325331.

  • 23.

    Liu Y. ; Qian L. H. Anhui Agri. Sci. Bull. 2015, 21, 2628.

  • 24.

    Liu L. J. ; Song W. F. Clin. Med. Engin. 2011, 18, 18571858.

  • 25.

    Xu W. ; Fu Z. Q.; Lin J.; Huang X. C.; Yu H. M.; Huang Z. H.; Fan S. M. Acta Pharmaceut. Sinica. 2014, 49, 17111717.

  • 26.

    Xu W. ; Fu Z.Q.; Lin J.; Huang X. C.; Chen D.; Yu H. M.; Huang Z. H.; Fan S. M. Chin. J. Chin. Mater. Med. 2014, 39, 43654372.

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