Quality Assessment of Panax notoginseng from Different Regions through the Analysis of Marker Chemicals, Biological Potency and Ecological Factors
Quality Assessment of Panax notoginseng from Different Regions through the Analysis of Marker Chemicals, Biological Potency and Ecological Factors
Hai-zhu Zhang 1 2 3
Da-hui Liu 1 3 4
Ding-kun Zhang 1 2 3
Yan-hui Wang 1 3
Gang Li 1 3
Gui- lin Yan 1 3
Li-juan Cao 1 3
Xiao-he Xiao 1 2 3
Lu-qi Huang 1 3
Jia-bo Wang 1 2 3
0 , Dong-cheng district, Beijing, 100062, China , 6 China Academy of Chinese Medical Sciences, National Resource Centre for Chinese Materia Medica , Beijing, 100700 , China
1 Funding: This research is supported by the National Natural Science Foundation of China , No. 81274026, 81403126
2 Chengdu University of Traditional Chinese Medicine , Chengdu, 611137, China , 2 China Military Institute of Chinese Medicine, 302 Military Hospital , Beijing, 100039, China , 3 Dali University , Dali, 671003 , China
3 Editor: Shilin Chen, Chinese Academy of Medical Sciences and Peking Union Medical College , CHINA
4 Yunnan Genuine Medicinal Materials Research and Development Centre, Kunming University of Science and Technology , Kunming, 650500, China, 5 China Medico Corporation, International building 811
Panax notoginseng (Burk.) F.H. Chen, called Sanqi in China, is a perennial herb that has been used as a medicinal herb in traditional Chinese medicine for more than 400 years. Because notoginseng is included in many proprietary Chinese medicines, the quality of notoginseng directly affects its efficacy and safety. However, considering the complex and special growth environment requirements of notoginseng, it is insufficient to evaluate its quality based solely on the analysis of marker chemicals. Thus, in this study, we tried to evaluate the quality of notoginseng with integrated indicators: (1) the concentration of five marker chemicals, notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1 and ginsenoside Rd; (2) the anticoagulant activity (ACA); and (3) twenty-one ecological factors (e.g., longitude, latitude, elevation and soil data). Using these 27 parameters, notoginseng from different regions could be distinguished effectively, indicating a remarkable divergence of quality. A correlation analysis showed that variations of the ecological factors were closely associated with the saponins content and biopotency. For instance, the total nitrogen (TN), alkali hydrolysis nitrogen (AHN) and rapidly available potassium (RAPT) were significantly correlated with ACA, and RAPT was significantly correlated with the content of ginsenoside Rd and notoginsenoside R1. The results demonstrated that the high-quality notoginseng was produced from the emerging regions such as Kunming, Qujing and Honghe, which had higher ACA and saponin content than the notoginseng pro-
Data Availability Statement: All relevant data are
within the paper and its Supporting Information
Competing Interests: The authors declare that no
competing interests exist. Three of the authors are
employed by a commercial company (China
Medico Corporation) and confirm this does not
duced in traditional regions such as Wenshan and Baise.
Panax notoginseng (Burk.) F.H. Chen, called sanqi in China, is a perennial herb that has been
used as a medicinal herb in traditional Chinese medicine for more than 400 years[
notoginseng belongs to the same genus as Chinese and Korean ginseng (Panaxginseng) and
American ginseng (Panax quinquefolium) [
Extensive phytochemical and pharmacological studies on this plant proved the
dammarane-type saponins to be the main bioactive principles [
], which are composed of a
protopanaxadiol and protopanaxatriol glycosides [
Currently, notoginseng is a commonly used for treating cardiovascular diseases, such as
lowering blood lipids, improving myocardial relaxation function, protecting arterial endothelium
from injury, blocking Ca2+ influx into VSMCs, and oestrogen-like activity[
is also used for inflammation, anti-inflammation in atherosclerotic lesion of the aorta, body
trauma, pain, and internal and external bleeding due to injury [
Because it has efficacy and lower adverse effects, notoginseng is gaining attention in Europe
and America. At present, the State Food and Drug Administration (SFDA) approved
production included 750 Panax notoginseng and Panax notoginseng drugs. Many proprietary Chinese
medicines contain notoginseng, including notoginseng tablets, notoginseng injury tablets,
compound Panax notoginseng, notoginseng tongshu capsules, and notoginseng yangxue capsules. In
recent years, the number of traditional medicines using notoginseng has been increasing.
Therefore, the quality of notoginseng medicinal materials is significant and will directly affect
the safety and efficacy in clinic. However, the quality of notoginseng is influenced by several
factors. The biological characteristics of notoginseng are determined by the special growth
environment and the growth cycle. The growing conditions require warm weather in winter and
cool weather in summer. The requirements for cold and heat, and moisture, are satisfied mostly
at low latitude and high elevation areas, so specific notoginseng is found in the Wenshan
Yunnan and Baise Guangxi geographic ranges. Among the two regions, Wenshan, where the drug
yield exceeds 90% of the total, has been nominated as ‘‘Sanqi Hometown” [
]. As a result
of the high medicinal value, the cultivated area in Wenshan and Baise, which are the
traditional regions, has continuously increased. However, the cultivation of notoginseng demands
specific soil, climate and geographical environment and more time than other herbs.
Prominent problems due to the successive cropping obstacles in the traditional regions are
increasingly, such as plant insect pests, soil rot, and the changing of nutrient, physical properties and
microflora in the soil [
], will significantly impact the quality of notoginseng.
However, there is no comprehensive method to evaluate the quality. The current quality
evaluation method measures the chemical composition, but there are numerous chemical
elements in the herb that are unknow or cannot be determined. When the contents of the
chemical components in different samples are the same, the biological activity can be significantly
different between samples[
], so there other components at work. Considering the complex
and special growth environment properties of notoginseng, and determining the chemical
composition cannot reflect the quality objectively and comprehensively, therefore, it is essential to
explore integrated evaluation methods.
In this study, we analysed the content of notoginsenoside R1, ginsenoside Rg1, ginsenoside
Re, ginsenoside Rb1, and ginsenoside Rd; calculated the anticoagulant activity (ACA); analysed
ecological factors, including longitude, latitude and elevation; and measured the pH, moisture,
organic content, total nitrogen (TN), alkali hydrolysis nitrogen (AHN), nitrate nitrogen (NN),
ammonia nitrogen (AN), total phosphorus (TP), rapidly available phosphorus (RAP), total
potassium (TPT), rapidly available potassium (RAPT), commutativity calcium (CCa),
commutativity magnesium (CMg), available iron (AFe), available manganese (AMn), available copper
2 / 12
(ACu), available zinc (AZn), and available boron (AB) in soil. We sought to analyse the
correlation between the producing area and the chemical constituents; analyse the biological activities
and ecological factors; comprehensively compared the quality characteristics of notoginseng to
explain the relationship between the chemical component content, biological activity and
ecological factors, to determine the decisive factors that affect the quality of notoginseng; and
provide integrated quality evaluation and a foundation for further search on the choice of a
suitable location to grow notoginseng, to guarantee its quality and clinical curative effect.
The samples of Panax notoginseng were collected from Guangxi and Yunnan Province in
China, cultivated three years (Table 1). The study was carried out on private land, we confirm
that the owner of the land gave permission to conduct the study on this site and the field
studies did not involve endangered or protected species.
All of the herbal samples were authenticated by Professor Xiaohe Xiao, and the voucher
specimens were deposited in People's Liberation Army 302 Hospital, Army Institute of
Traditional Chinese Medicine (TCM), Beijing, China.
The chromatographic experiments were performed on an Agilent 1120 series HPLC system
(Agilent Technologies Inc., Shanghai, China). The separation was conducted at 30°C on an
Agilent Zorbax eclipse Shim-pack PREP-ODS(H) kit column (4.6 × 250 mm, 5 μm). The
mobilephase consisted of solvent A (acetonitrile) and solvent B (water) flowing at 1 mL min−1. The
initial conditions were 18% A for 12 min, and a linear gradient was performed to increase from
18% A to 38% A within 23 min, and then 38% A within 7 min, which was held for 0.5 min
before returning to 18% A. The scan range for PDA was 203 nm with a sample size 20 μL.
The five reference compounds, notoginsenoside R1 (110745–200617), ginsenoside Rg1
(110703–201027), ginsenoside Rb1 (110704–201122), ginsenoside Rd (111818–201001) and
ginsenoside Re (110754–200822), were accurately weighed: 1mg was dissolved in a 10 mL
volumetric flask with methanol to produce standard stock solutions. Samples of herbal materials
were ground into a fine powder and then passed through a 20 mesh (0.9 mm) sieve. Sample
powder (0.3 g) was accurately weighed and transferred into a 50 mL triangle flask. Then,
100% methanol (25 mL) was added, the samples were stored over- night, and were then
ultrasonicated for 30 min. When cool, methanol was added to compensate for weight loss.
After filtering through a 0.45μm filter membrane, the filtrate was ready to be used.
Biopotency assay experiments
The standard compound, aspirin, was accurately weighed: 50 mg was dissolved in a 10 mL
volumetric flask with physiological saline to produce standard stock solutions, and the defined
biological value was 500U g-1. Samples of herbal materials were ground into fine powder and
passed through a 20 mesh (0.9 mm) sieve. Sample powder (10 g) was accurately weighed and
transferred into a 250 mL triangle flask. Deionized water (100 mL) was added, and the samples
were allowed to sit for 30 min, before being ultrasonicated twice for 30 min. Samples were then
merged, enriched, decompression dried, and prepared with normal saline solution to produce
sample stock solutions at 15mg mL-1. Ear marginal blood was collected by needle from two
New Zealand rabbits (Males 2.5–4 kg) (SCXK 2011–0004), that were fasting 12 h. Blood was
transferred into a sodium citrate anticoagulation tube, centrifuged for 15 min at 3000 r/min,
and the plasma was stored at 4°C. The plasma and 200 μL of sample or standard stock solution
were mixed at 37°C for 180 s, and the activated partial thromboplastin time (APTT) was
detected by an automatic coagulation analyser (ACA)(Sysmex CA-7000, Japan). The
experimental procedures and the animal use and care protocols were approved by the Committee on
Ethical Use of Animals of 302 Military Hospital of China.
Soil samples were obtained from 0 to 15 cm soil layer. The surface mulch was removed, and the
samples were placed in bags. The soil samples were dried in the shade to a constant weight
under natural conditions and were passed through a 20 mesh and 100 mesh sieve.
pH was measured by the potentiometric method (water: soil = 2.5:1); the organic matter
content was measured using the potassium dichromate volumetric method; total nitrogen
(TN) was determined by Kjeldahl nitrogen distillation; alkali hydrolysis nitrogen (AHN, nitrate
nitrogen (NN), and ammonia nitrogen (AN) were measured using the alkali solution diffusion
method; total phosphorus (TP) was determined using the sodium hydroxide
melting—molybdenum antimony colorimetric method; rapidly available phosphorus (RAP) was determined
using the Olsen method; total potassium (TPT) was measure by sodium hydroxide melting—
atomic absorption spectrophotometry; rapidly available potassium (RAPT) was measured
using ammonium leaching acetic acid—atomic absorption spectrophotometry; changeable
calcium (CCa) and magnesium (CMg) were determined using the EDTA—acetic acid ammonium
exchange method; and available iron, manganese, copper, zinc, and boron (AFe, AMn, ACu,
AZn, AB) were measured DTPA extraction—atomic absorption spectrophotometer.
3. Results and Discussion
Analysis of the chemical compounds
The linearity, regression, and linear ranges of five investigated compounds were determined by
HPLC. The data indicated a good relationship between the concentrations and peak areas of
the compounds within the test ranges (R2 0.9990). The LOQs and LODs of all compounds
were less than 6.15 and 13.17μg mL−1 (Table 2). The overall RSDs of the intra- and inter-day
variations for analytes were not more than 2.11% and 11.54%, respectively. The established
method also had acceptable accuracy, with spike recovery of 98.31–103.57% for all analytes.
For the stability test, the RSDs of the peak areas for compounds detected within 12 h were
lower than 2.59%. These results demonstrated that the HPLC method was linear, sensitive,
4 / 12
Y = 1E+06X-1075.1
Y = 1E+06X+302.9
Y = 1E+06X-2850.8
Y = 1E+06X-2187.4
Y = 1E+06X-671.9
Analysis of the bioactivity of Panax notoginseng
The analysis of the anticoagulant activity (ACA) used the quantum parallel reaction method of
the State Pharmacopoeia Committee of Establishment of China Pharmacopoeia of Bioassay
Statistical Program BS2000 technology. The biological reliability verification results are shown
in Table 3 and Fig 2. The regression had a significant difference (P < 0.01), indicating that
APTT increased regularly with the dose increase. The agent had a significant difference
(P < 0.01), indicating that the test dose rate and test arrangement were reasonable. Deviation
from parallel had no significant difference (P > 0.05), suggesting a parallel linear relationship
between the standard group (S) and the test group (T). The ACA was from 89.47 to 218.87U
g−1 in ten batches from different origin samples, and varied by nearly three times in different
origin samples which indicated that the sample origin had a strong influence on the quality of
notoginseng medicinal materials. For example, Xundian County in Kunming (sample No. 8),
Fig 1. Chromatogram. (A) mixed standard solution. (B) sample solution. (C) content determination results.
5 / 12
Degrees of freedom
Sum of squares of deviations
Analysis of soil characteristics
Fifteen soil characteristics, including TN, AHN, NN, AN, TP, RAP, TPT, RAPT, CCa, CMg,
AFe, AMn, ACu, AZn, and AB in 10 batches of different regions of notoginseng were analysed
as shown in Table 4. The producing region in Guandu County in Kunming (sample No. 7) had
the highest AHN, TN, NN, and RAPT. N and K were the main nutrient elements of
notoginseng. The application of N and K fertilizer was an important method to ensure the quality of
]. Kunming Region exhibited good quality soil owing to the high soil N and
The correlation analysis
From the data on the correlation coefficient among the main chemical compounds, and the
bioactivity and ecological factors of notoginseng in Figs 3 and 4, we can see that
notoginsenoside R1, ginsenoside Rd and anticoagulant activity(ACA) were significantly negatively
correlated with the longitude and were significantly positively correlated with the latitude and
elevation. Notoginsenoside R1 and ginsenoside Rd were significantly positively correlated with
ACA (P<0.05), which indicates that the higher the content of notoginsenoside R1 and
ginsenoside Rd, the higher the ACA. Notoginsenoside R1 was positively correlated with the TN, AHN,
TP and RAPT. Ginsenoside Rd was positively correlated with AFe. Ginsenoside Rb1 was
significantly positively correlated with TPT (P<0.05). Ginsenoside Rd was significantly positively
Fig 2. The result of the anticoagulant activity (ACA) analysis of notoginseng.
6 / 12
Fig 3. Correlation analysis. (A-E)The correlation between the content of chemical compounds (notoginsenoside R1, ginsenoside Rg1, ginsenoside Re,
ginsenoside Rb1, ginsenoside Rd) and ecological factors. (F)The correlation between bioactivity and ecological factors. (G)The correlation between the
content of chemical compounds and bioactivity.
Fig 4. Thermograph of the correlation analysis of all factors.
8 / 12
Fig 5. (A)PLS-DA of the producing regions versus the chemical constituents, biological activities and ecological factors. (B)The v-plot of the variables to
distinguish between traditional regions (TR) and emerging regions (ER). (C-K) The decisive quality characteristics comparison of notoginseng from TR
correlated with AMn and RAPT (P<0.05). Ginsenoside Re was significantly negatively
correlated with TN and AHN (P<0.05). The anticoagulant activity was significantly positively
correlated with TN, ANH, RAPT and ginsenoside R1(P<0.05). Fig 4 presents the positive
correlation relationship between the several factors, including ACA, TN, AHN, RAPT, R1,
elevation and latitude. Re and longitude had a negative correlation relationship with the other
factors. Fig 5 shows that the notoginseng samples were divided into 2 main clusters in the
PLS-DA. The cultivated regions in Wenshan and Baise, which were the Sanqi hometown, were
defined as the traditional regions (TR), and the regions in Kunming, Honghe and Qujing were
defined as the emerging regions (ER). Such division indicated that different producing regions
could significantly distinguish from TR to ER by the different chemical constituents, biological
activities and ecological factors of notoginseng. The v-plot results of the variables to distinguish
between TR and ER are shown in Fig 5. The ACA, latitude, elevation, longitude, TN, AHN, Re
and longitude were the important factors in distinguishing between TR and ER. Fig 5 C-K
shows an increasing tendency in latitude, elevation, TN, AHN, ACA, and RAPT, and a
decreasing tendency in longitude and the content of Re from TR to ER. Therefore, the higher latitude,
elevation, TN, AHN, ACA, and RAPT and lower of longitude and Re were the main reasons
that the quality characteristics of TR cultivated notoginseng were worse than ER cultivation.
The production of high-quality notoginseng has been migrating from TR, which has been
cultivated hundreds years, to ER in recent decades. The prominent problems due to the successive
cropping obstacles in the TR, such as plant insect pests, soil rot, and the changing of nutrient,
physical properties and microflora in soil are increasing. The ER, has the superior specific soil,
climate and geographical environment to TR, resulting in high-quality notoginseng.
The quality of notoginseng medicinal materials is not only affected by the chemical
constituents. In this study, a reliable quality assessment through the analysis of marker chemicals,
9 / 12
biological potency and ecological factors was established for Panax notoginseng from different
regions. We found that high-quality notoginseng was produced by emerging regions such as
Kunming, Qujing and Honghe, which had higher ACA and saponin content than the
notoginseng produced in traditional regions such as Wenshan and Baise.
S1 File. Approval of Experimental Animal Welfare and Ethics.
S1 Table. The results of component content determination.
This research is supported by the National Natural Science Foundation of China (No.
Conceived and designed the experiments: HZZ JBW XHX LQH.
Performed the experiments: HZZ JBW YHW DHL XHX DKZ LQH.
Analyzed the data: DKZ HZZ GL GLY LJC.
Contributed reagents/materials/analysis tools: JBW YHW DHL LQH XHX.
Wrote the paper: HZZ DHL JBW LQH.
10 / 12
11 / 12
1. Zhang Y , Han LF , Sakah KJ , Wu ZZ , Liu LL , Agyemang K , et al. Bioactive Protopanaxatriol Type Saponins Isolated from the Roots of Panax Notoginseng (Burk .) F. H. Chen . Molecules. 2013 ; 18 : 10352 ± 10366 . doi: 10 .3390/molecules180910352 PMID: 24064450
Wang D , Koh HL , Hong Y , Zhu HT , Xu M , Zhang YJ , et al. Chemical and morphological variations of Panax notoginseng and their relationship . Phytochemistry . 2013 ; 93 : 88 ± 95 . doi: 10 .1016/j.
phytochem. 2013 . 03 .007 PMID: 23566718
3. Chan EC , Yap SL , Lau AJ , Leow PC , Toh DF , Koh HL . Ultra-performance liquid chromatography/timeof-flight mass spectrometry based metabolomics of raw and steamed Panax notoginseng , Rapid Commun Mass Spectrom . 2007 ; 21 : 519 ± 528 . doi: 10 .1002/rcm.2864 PMID: 17238214
Wan JB , Li SP , Chen JM , Wang YT . Chemical characteristics of three medicinal plants of the Panax genus determined by HPLC-ELSD . J. Sep. Sci . 2007 ; 30 : 825 ± 832 . PMID: 17536727
5. Xu QF , Fang XL , Chen DF . Pharmacokinetics and bioavailability of ginsenoside Rb1 and Rg1 from Panax notoginseng in rats . J Ethnopharmacol . 2003 ; 84 : 187 ± 192 . doi: 10 .1016/S0378- 8741 ( 02 ) 00317 - 3 PMID: 12648814
6. Han JA , Hu WY , Sun ZH . Effect of Panax notoginseng Saponin on Ca2+, CaM in craniocerebral injury . Chin J Integr Tradit West Med 1999 ; 19 : 227 ± 9 . PMID: 11783273
7. Li XH , Li SH . Effects of total saponins of Sanchi (Panax pseudo-ginseng notoginseng) on TNF, NO and its mechanisms . Chin Tradit Herbal Drugs . 1999 ; 30 : 514 ± 7 .
8. Ma LY , Xiao PG . Effects of saponins of Panax notoginseng on intracellular free Ca2+ concentration in dissociated neurons . Chin Pharm J . 1998 ; 33 : 467 ± 9 .
9. Ma LY , Xiao PG , Liang FQ , Wu JH . Protective effects of Panax notoginseng saponins on primary cortical cultures of rat . Chin Pharm J 1998 ; 33 : 143 ± 5 .
10. Zhang GQ , Ye RG , Kong QY , Yang NS , Zhang JL , Guan WM , et al. Panax notoginseng saponins induced of human renal interstitial fibroblasts and its mechanisms . Chin J Nephrol 1998 ; 14 : 93 ± 5 .
11. Zhu XX , Mao YW , He RX , Yamamoto A , Shoyama Y. Determination of ginsenosides in Panax genseng by HPLC . Chin J Biochem Pharm 1998 ; 19 : 28 ± 30 .
12. Du QZ , Jerz G , Waibel R , Winterhalter P. Isolation of dammarane saponins from Panax notoginseng by high-speed countercurrent chromatography . J Chromatogr A 2003 ; 1008 : 173 ± 80 . PMID: 12967182
13. Lau AJ , Woo SO , Koh HL . Analysis of saponins in raw and steamed Panax notoginseng using highperformance liquid chromatography with diode array detection . J Chromatogr A 2003 ; 1011 : 77 ± 87 . doi: 10 .1016/S0021- 9673 ( 03 ) 01135 -X PMID: 14518765
14. Sun HX , Yang ZG , Ye YP , Structure and biological activity of protopanaxatriol-type saponins from the roots of Panax notoginseng , International Immunopharmacology . 2006 ; 6 : 14 ± 25 . doi: 10 .1016/j. intimp. 2005 . 07 .003 PMID: 16332509
15. Ling S , Nheu L , Dai A , Guo Z , Komesaroff P , Effects of four medicinal herbs on human vascular endothelial cells in culture . Int. J.Cardiol . 2008 ; 128 : 350 ± 358 . doi: 10 .1016/j.ijcard. 2007 . 05 .111 PMID: 17692965
16. Ji W , Gong BQ , Hypolipidemic effects and mechanisms of Panax notoginseng on lipid profile in hyperlipidemic rats . J Ethnopharmacol . 2007 ; 113 : 318 ± 324 . doi: 10 .1016/j.jep. 2007 . 06 .022 PMID: 17681443
17. Xu L , Liu JT , Liu N , Lu PPP , Pang XM , Effects of Panax notoginseng saponins on proliferation and apoptosis of vascular smooth muscle cells , Journal of Ethnopharmacology . 2011 ; 137 : 226 ± 230 . doi: 10 . 1016/j.jep. 2011 . 05 .020 PMID: 21619919
18. Chan P , Thomas GN , Tomlinson B , Protective effects of trilinolein extracted from Panax notoginseng against cardiovascular disease . Acta Pharmacologica Sinica . 2002 ; 23 : 1157 ± 1162 . PMID: 12466054
19. Cicero AF , Vitale G , Savino G , Arletti R , Panax notoginseng effects on fibrinogen and lipid plasma level in rats fed on a high-fat diet . Phytother Res . 2003 ; 17 : 174 ± 178 . doi: 10 .1002/ptr.1262 PMID: 12601683
20. Jia Y , Li XH , Liu Y , Zhang HG , Atherosclerosis lesion is accelerated by persistent systemic inflammation but attenuated by saponins from Panax notoginseng in rabbits . Journal of Medical Colleges of PLA . 2008 ; 23 : 38 ± 44 .
21. Chen SW , Li XH , Ye KH , Jiang ZF , Ren XD , Total saponins of Panax notoginseng protected rabbit iliac artery against balloon endothelial denudation injury . Acta Pharmacologica Sinica . 2004 ; 25 : 1151 ± 1156 . PMID: 15339390
22. Guan YY , Zhou JG , Zhang Z , Wang GL , Cai BX , Hong L , et al, Ginsenoside-Rd from Panax notoginseng blocks Ca2+ influx through receptorand store-operated Ca2+ channels in vascular smooth muscle cells . European Journal of Pharmacology 2006 ; 548 : 129 ± 136 . doi: 10 .1016/j.ejphar. 2006 . 08 .001 PMID: 16973156
23. Chan RY , Chen EF , Dong A , Guo A , Wong MS ,. Estrogen-like activity of ginsenoside Rg1 derived from Panax notoginseng . J Clin Endocrinol Metab . 2002 ; 87 : 3691 ± 3695 . doi: 10 .1210/jcem.87.8.8717 PMID: 12161497
24. Dong TT , Cui XM , Song ZH , Zhao KJ , Ji ZN , Lo CK , et al. Chemical assessment of roots of Panax notoginseng in China: regional and seasonal variations in its active constituents . J Agric Food Chem . 2003 ; 51 : 4617 ± 23 . doi: 10 .1021/jf034229k PMID: 14705886
25. Wei JX , Du YC . Modern science research and application of Panax Notoginseng. Kunming 7 Yunnan Science and Technology Press; 1996 . p. 426 .
26. Cicero AF , Vitale G , Savino G , Arlett R , Panax notoginseng (Burk.) effects on fibrinogen and lipid plasma level in rats fed on a high-fat diet . Phytother Res . 2003 ; 17 : 174 ±8. doi: 10 .1002/ptr.1262 PMID: 12601683
27. Zheng GZ , Yang CR . Biology of Panax notoginseng and its application . Beijing 7 Science Press; 1994 .
28. Ma WG , Mizutani M , Malterud KE , Lu SL , Ducrey B , Tahara S , et al. Saponins from the roots of Panax notoginseng . Phytochemistry . 1999 ; 52 : 1133 ±9. doi: 10 .1016/S0031- 9422 ( 99 ) 00364 - 7
29. Liu Y , Xie MX , Kang J , Zheng D , Studies on the interaction of total saponins of Panax notoginseng and human serum albumin by Fourier transform infrared spectroscopy . Spectrochim Acta . 2003 ; 59 : 2747 ± 58 . doi: 10 .1016/S1386- 1425 ( 03 ) 00055 - 6 PMID: 14499835
30. An N , Cui XM , Xiao FH , Chen ZJ , Duan CL , Dynamic studies on physiological and biochemical changes during fruit development of Panax notoginseng . Chin. Tradit. Herbal Drugs . 2006 ; 37 : 1086 ± 1088 (in chinese).
31. Guo XC , 2007 . New green industryÐtakes Wenshan Panax notoginseng status and its future as an example . Ecol. Econ . 1 : 114 ± 117 .
32. Cui XM , Huang LQ , Guo LP , Liu DH . Chinese Sanqi industry status and development countermeasures , Chinese journal of traditional Chinese medicine (TCM) . 2014 ; 39 : 553 ± 556 . PMID: 25204122
33. Liu LL , Zhao AJ , Yang Y , Jin H , Cui XM , Liu DH , Comparative Analysis of Physical and Chemical Properties of Panax notoginseng replant Soils in Different Intervals . Southwest China Journal of Agricultural Sciences . 2013 ; 26 : 1946 ± 1952 (in chinese).
34. Marschner P , Crowley DE , Yang CH . Development of specific rhizosphere bacterial communities in relation to plants pecies,nutrition and soil type . Plant and Soil . 2004 ; 261 : 199 ± 208 .
35. Mitchell L , Cheang SK , Gerald M , Suresh G , Nikolaus JS . An in vitro study of anti-inflammatory activity of standardised Andrographis paniculata extracts and pure andrographolide . Complementary and Alternative Medicine . 2015 ; 15 : 18 . doi: 10 .1186/s12906-015 -0525-7 PMID: 25888070
36. Huang CM , Cui XM , Lan L , Chen WD , Wang CX , Yang XY , et al. Research on output and quality of Panax notoginseng and annual change characteristics of N, P and K nutrients of planting soil under stereo-cultivation , China Journal of Chinese Materia Medica . 2015 ; 40 : 2930 ± 6 . PMID: 26677689
37. Zheng DM , Wang L , Ou XH , Guo LP , Hao QX , Liu DH , et al. Comparison of agronomic traits of Panax notoginseng between traditional cultivated fields and new cultivated fields . China Journal of Chinese Materia Medica . 2014 , 39 : 558 ± 65 . PMID: 25204123
38. Liu DH , Wang L , Cui XM , Guo LP , Jin H , Zhu XY , et al. Study on dynamic change law of N, P and K in Panax notoginseng plant soils with different interval year . China Journal of Chinese Materia Medica . 2014 , 39 : 572± 9 . PMID: 25204125