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Journal of Ginseng Research JGR149_proof ■ 24 December 2015 ■ 1/13 J Ginseng Res xxx (2015) 1e13 55 Contents lists available at ScienceDirect 56 57 Journal of Ginseng Research 58 59 60 journal homepage: http://www.ginsengres.org 61 62 63 Research article 64 65 1 Rapid characterization of ginsenosides in the roots and rhizomes of 66 2 67 3 Panax ginseng by UPLC-DAD-QTOF-MS/MS and simultaneous 68 4 determination of 19 ginsenosides by HPLC-ESI-MS 69 5 70 6 1 1 1,* 2 3 71 7 Q14 Hong-Ping Wang , You-Bo Zhang , Xiu-Wei Yang , Da-Qing Zhao , Ying-Ping Wang 72 8 1 State Key Laboratory of Natural and Biomimetic Drugs, Department of Natural Medicines, School of Pharmaceutical Sciences, Peking University, Beijing, 73 9 China 74 2 10 Changchun University of Chinese Medicine, Changchun, China 75 3 Institute of Special Wild Economic Animals and Plants Science, Chinese Academy of Agricultural Sciences, Changchun, China 11 76 12 77 13 article info abstract 78 14 79 15 80 16 Article history: Background: Ginsenosides are the characteristic and principal components which manifest a variety of Received 10 August 2015 81 17 the biological and pharmacological activities of the roots and rhizomes of Panax ginseng (GRR). This study Received in Revised form was carried out to qualitatively and quantitatively determine the ginsenosides in the cultivated and 82 18 28 November 2015 forest GRR. 83 Accepted 1 December 2015 19 Methods: A rapid and sensitive ultra-high-performance liquid chromatography coupled with diode-array 84 Available online xxx 20 detector and quadrupole/time of flight tandem mass spectrometry (UPLC-DAD-QTOF-MS/MS) was 85 21 applied to the qualitative analysis of ginsenosides and a 4000 QTRAP triple quadrupole tandem mass 86 22 Keywords: cultivated ginseng spectrometer (HPLC-ESI-MS) was applied to quantitative analysis of 19 ginsenosides. 87 23 forest ginseng Results: In the qualitative analysis, all ingredients were separated in 10 min. A total of 131 ginsenosides 88 24 HPLC-ESI-MS were detected in cultivated and forest GRR. The method for the quantitative determination was validated 89 fi 25 Panax ginseng for linearity, precision, and limits of detection and quanti cation. 19 representative ginsenosides were 90 UPLC-DAD-QTOF-MS/MS quantitated. The total content of all 19 ginsenosides in the forest GRR were much higher than those in the 26 91 27 cultivated GRR, and were increased with the growing ages. Conclusion: This newly developed analysis method could be applied to the quality assessment of GRR as 92 28 well as the distinction between cultivated and forest GRR. 93 29 Copyright Ó 2015, The Korean Society of Ginseng, Published by Elsevier. All rights reserved. 94 30 95 31 96 32 97 33 1. Introduction natural populations of ginseng to such a degree that it has become 98 34 threatened with extinction in certain regions. It is necessary to 99 35 Asian ginseng, Panax ginseng Meyer, is a deciduous perennial cultivate the most commonly used ones to guarantee supplies. 100 36 herb. It belongs to the family Araliaceae, which is distributed in During the long-term natural and artificial selection, three culti- 101 37 Northeast China, Korea and the Russian Far East. The roots and vated types have formed, namely garden ginseng, forest ginseng 102 38 rhizomes of ginseng (GRR) are known as the lord or king of herbs. and transplanted wild ginseng. Garden ginseng is produced as a 103 39 This drug has been an important component of Chinese medicine type grown purely under artificial conditions, and its growth usu- 104 40 for over 3,000 yr and is now widely used around the world [1]. ally spans only 4e7 yr. Forest ginseng is developed by sowing seeds 105 41 Ginseng is also becoming popular in the public food field. It has of garden ginseng into natural environments and letting them grow 106 42 been approved by the Chinese government as a new food resource without any artificial disturbance or management, and its growth 107 43 in 2012 (http://news.xinhuanet.com/fortune/2012-09/05/c_ usually spans over 10 yr. Transplanted wild ginseng is domesticated 108 44 112970866.htm). Nowadays, wild harvest has depleted the by transplanting seedlings of wild ginseng into artificial or 109 45 110 46 111 47 * 112 48 Corresponding author. State Key Laboratory of Natural and Biomimetic Drugs and Department of Natural Medicines, School of Pharmaceutical Sciences, Peking University, Beijing 100191, People’s Republic of China. 113 49 E-mail address: [email protected] (X.-W. Yang). 114 50 115 51 This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) 116 which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. 52 117 53 p1226-8453 e2093-4947/$ e see front matter Copyright Ó 2015, The Korean Society of Ginseng, Published by Elsevier. All rights reserved. 118 54 http://dx.doi.org/10.1016/j.jgr.2015.12.001 119 Please cite this article in press as: Wang H-P, et al., Rapid characterization of ginsenosides in the roots and rhizomes of Panax ginseng by UPLC- DAD-QTOF-MS/MS and simultaneous determination of 19 ginsenosides by HPLC-ESI-MS, Journal of Ginseng Research (2015), http://dx.doi.org/ 10.1016/j.jgr.2015.12.001 JGR149_proof ■ 24 December 2015 ■ 2/13 2 J Ginseng Res 2015;-:1e13 1 semiartificial environments. Because forest ginseng may become an total ion current (TIC) of total ginsenosides. So the ginsenosides 66 2 alternative source of wild ginseng, China and Korea are vigorously detected were limited and the characterization of 28 ginsenosides 67 3 developing forest ginseng. Ginsenosides [2] are the characteristic gave the perfect results. Usually, ginsenosides in minor or trace 68 4 and principal components which manifest a variety of the biolog- amounts cannot be detected. Otherwise, the analytical time is very 69 5 ical and pharmacological activities of GRR [3e5] and have been an long, which is not convenient to rapidly qualify the ginsenosides in 70 6 important index in assessing the quality of GRR and its products [6]. ginseng. In order to rapidly clarify the basic chemical substances of 71 7 Naturally occurring ginsenosides can be further classified into three GRR, a rapid and sensitive method, which can thoroughly detect the 72 8 major types, namely types of protopanaxatriol (PPT), proto- main and minor or trace amounts of ginsenosides, should be 73 9 panaxadiol (PPD) and oleanolic acid (OA), according to their sapo- established. In the present study, a new rapid and sensitive ultra- 74 10 genins with a dammarane or oleanane skeleton (Fig. 1). Many high-performance liquid chromatography coupled with diode- 75 11 analytical approaches have been developed to quantify ginseno- array detector and quadrupole/time of flight tandem mass spec- 76 12 sides, including TLC [7], HPLC coupled with a UV detector [8,9] or an trometry (UPLC-DAD-QTOF-MS/MS) method was established to 77 13 evaporative light scattering detector (ELSD) [10e12], and high- identify the basic chemical substances. As a result, a total of 131 78 14 performance LC-MS [13]. Because of the diversity, similarity and ginsenosides were characterized. Also, a sensitive and practical 79 15 complexity of the chemical structures, the analysis of ginsenosides HPLCÀMS/MSn method was developed to simultaneously deter- 80 16 is a great challenge. Liquid chromatography coupled with electro- mine 19 ginsenosides in GRR for the first time. This newly devel- 81 17 spray ionization tandem mass spectrometry (LC-ESI-MS/MS) is a oped qualitative and quantitative method could be applied to the 82 18 powerful tool for the ginsenosides analysis. Song et al [14] have holistic quality assessment of GRR. 83 19 identified three pairs of ginsenoside (G) isomers (G-Rg2 and G-Rg3, 84 20 G-Rg1 and G-F11 as well as G-Rd and G-Re) and Miao et al [15] have 85 21 studied the fragmentation pathway of 9 ginsenosides, namely G- 2. Materials and methods 86 22 Rb1, Rb2, Rc, Rd, Re, Rf, Rg1, Rg2, and F11 by LC-MS/MS. Because MS 87 23 can provide the information of molecular formula and fragmenta- 2.1. GRR samples 88 24 tion ions, some researchers have identified ginsenosides in red 89 25 ginseng by LC-ESI-MS/MS methods. For instance, Zhang et al [16] All GRR samples are listed in Table 1. The botanical origins of 90 26 characterized 25 ginsenosides in 152 min while Xie et al [9] iden- samples were identified by Professor Da-Qing Zhao, of the Chang- 91 27 tified 28 ginsenosides in 80 min. In these reports, the methods chun University of Chinese Medicine, China. A voucher specimen 92 28 established were suitable for the analysis of the main ion peaks in has been deposited in the State Key Laboratory of Natural and 93 29 94 30 95 31 96 32 97 33 98 34 99 35 100 36 101 37 102 38 103 39 104 40 105 41 106 42 107 43 108 44 109 45 110 46 111 47 112 48 113 49 114 50 115 51 116 52 117 53 118 54 119 55 120 56 121 57 122 58 123 59 124 60 125 61 126 62 127 63 128 64 129 65 Fig. 1. The chemical structures of 19 reference standards. 130 Please cite this article in press as: Wang H-P, et al., Rapid characterization of ginsenosides in the roots and rhizomes of Panax ginseng by UPLC- DAD-QTOF-MS/MS and simultaneous determination of 19 ginsenosides by HPLC-ESI-MS, Journal of Ginseng Research (2015), http://dx.doi.org/ 10.1016/j.jgr.2015.12.001 JGR149_proof ■ 24 December 2015 ■ 3/13 H.-P.
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