Metabolomic Approach for Discrimination of Four- and Six-Year-Old Red Ginseng (<i>Panax ginseng</i>) Using UPLC-QToF-MS

Abstract

Panax ginseng C.A. MEYER is one of the most popular medicinal herbs in Asia and the chemical constituents are changed by processing methods such as steaming or sun drying. Metabolomic analysis was performed to distinguish age discrimination of four-and six-year-old red ginseng using ultra-performance liquid chromatography quadruple time of flight mass spectrometry (UPLC-QToF-MS) with multivariate statistical analysis. Principal component analysis (PCA) showed clear discrimination between extracts of red ginseng of different ages and suggest totally six discrimination markers (two for four-year-old and four for six-year-old red ginseng). Among these, one marker was isolated and the structure determined by NMR spectroscopic analysis was 13-cis-docosenamide (marker 6-1) from six-year-old red ginseng. This is the first report of a metabolomic study regarding the age differentiation of red ginseng using UPLC-QToF-MS and determination of the structure of the marker. These results will contribute to the quality control and standardization as well as provide a scientific basis for pharmacological research on red ginseng.Key words Panax ginseng; red ginseng; multivariate analysis; ultra-performance liquid chromatography quadruple time of flight mass spectrometry (UPLC-QToF-MS); metabolomics; 13-cis-docosenamide Korean ginseng (Panax ginseng C.A. MEYER) is one of the most widely used and acclaimed herbs in the world.1-3) Traditionally, the root of P. ginseng, the most used and valuable part, is physically subdivided into three groups; the main root, lateral root, and root hairs.1) It has already been reported that the chemical constituents and efficacy of each part of the ginseng root are quite different.1) It has been processed to make white ginseng (by air-drying the roots after peeling or not peeling) and red ginseng (by steaming the roots at 98-100°C without peeling) to enhance its preservation and efficacy. Red ginseng is more common as an herbal medicine, because steaming induces changes in the chemical constituents and enhances the biological activities of ginseng. 4)P. ginseng is generally cultivated for four or six years in the field before harvest. Thus, four and six year cultivated P. ginseng is a common item in the Korean ginseng market. However, six year cultivated P. ginseng and products thereof (including white ginseng and red ginseng) are produced and consumed much more than others. Because the cultivation age and harvest season have a significant effect on the quality and efficacy of ginseng, products of six year cultivated P. ginseng are also more expensive than products of four year cultivated P. ginseng. Additionally, a low survival rate and higher cultivation costs contribute to the expensive price of six year cultivated P. ginseng. The accurate determination of the cultivation age of ginseng is a very important problem in the market, although the cultivation age of ginseng can hardly be determined by the physical appearance alone, such as by the number of stem vestiges in rhizome. 5)In recent years, ...

A Metabolomic Approach for the Discrimination of Red Ginseng Root Parts and Targeted Validation

“…We tried to identify all of the markers of the main roots and were able to confirm the marker ion g in gingerglycolipid B via comparison with another publication [ 11 ]. Additionally, marker ion l was 13- cis -docosenamide, which was the same compound as that found in our previous report [ 12 ]. The others (ion h and k ), owing to their lower amounts, did not result in anything.…”

“…In the case of the fine root samples, five marker ions were confirmed to be ginsenoside ( a , ginsenoside Rd; b , ginsenoside Rc; c , ginsenoside Rb2; d , ginsenoside Rb1; f , ginsenoside 20( S )-Rg2). Moreover, Marker Ion l was 13- cis -docosenamide, which was identified as a marker for six-year-old RG in our previous research [ 12 ]. Isolation and spectroscopic identification were performed to identify Remaining Marker Ions g , h , and k .…”

Qualitative and quantitative analysis of saponins in the flower bud of Panax ginseng (Ginseng Flos) by UFLC‐Triple TOF‐MS/MS and UFLC‐QTRAP‐MS/MS

“…. Fifty‐one saponins were identified in GF by comparison of MS data and retention behavior found in the literature and are shown in Table . Twenty‐one compounds (Notoginsenoside R 1 , R 2 , and ginsenoside Re, Rg 1 , Rb 1 , Rf, 20(S)‐Rg 2 , 20(R)‐Rg 2 , 20(R)‐Rh 1 , Rc, Ro, Rb 3 , Rb 2 , F 1 , Rk 3 , Rd, F 4 , F 2 , 20(S)‐Rg 3 , 20(R)‐Rg 3 , and Chikusetsusaponin IVa) were also characterized by the assistance of their standards.…”

“…However, compared with HPLC, the ultra‐fast liquid chromatography coupled with triple quadrupole‐linear ion trap tandem mass spectrometry (UFLC‐QTRAP‐MS/MS) method is more efficient for the analysis of TCM because it can solve the co‐outflow of complex components, which has superiorities of high sensitivity, good separation efficiency, and high peak capacity. Moreover, ultra‐fast liquid chromatography with triple quadrupole‐time of flight tandem mass spectrometry (UFLC‐Triple TOF‐MS/MS) method established in this study can provide structural information of the samples . In this study, 51 saponins in GF were identified by UFLC‐Triple TOF‐MS/MS, furthermore, an efficient UPLC‐QTRAP‐MS/MS method was developed to determine the contents of 12 ginsenosides in GF, including protopanaxatriol‐type ginsenosides (ginsenoside Rd, Rb 2 , Rc, Rb 1 , and F 2 ), protopanaxadiol‐type ginsenosides (ginsenoside Re, Rg 1 , Rf, 20(S)‐Rg 2 , 20(R)‐Rg 2 , and F 1 ), and oleanolic acid‐type ginsenosides (ginsenoside Ro).…”

“…Moreover, ultra-fast liquid chromatography with triple quadrupole-time of flight tandem mass spectrometry (UFLC-Triple TOF-MS/MS) method established in this study can provide structural information of the samples. [14][15][16][17][18] In this study, 51 saponins in GF were identified by UFLC-Triple TOF-MS/MS, furthermore, an efficient UPLC-QTRAP-MS/MS method was developed to determine the contents of 12 ginsenosides in GF, including protopanaxatriol-type ginsenosides (ginsenoside Rd, Rb 2 , Rc, Rb 1 , and F 2 ), protopanaxadiol-type ginsenosides (ginsenoside Re, Rg 1 , Rf, 20(S)-Rg 2 , 20(R)-Rg 2 , and F 1 ), and oleanolic acid-type ginsenosides (ginsenoside Ro). These findings are beneficial to the comprehensive evaluation and control of the internal quality of GF and its relevant products.…”