Methanol-involved heterogeneous transformation of ginsenoside Rb1 to rare ginsenosides using heteropolyacids embedded in mesoporous silica with HPLC-MS investigation

Mengya Zhao,Yusheng Xiao,Yanyan Chang,Lu Tian,Yujiang Zhou,Shuying Liu,Huanxi Zhao,Yang Xiu 고려인삼학회 2024 Journal of Ginseng Research Vol.48 No.4

Background: The biological activity and pharmacological effects of rare ginsenosides have been proven to besuperior to those of the major ginsenosides, but they are rarely found in ginseng. Methods: Ginsenoside Rb1 was chemically transformed with the involvement of methanol molecules by a synthesizedheterogeneous catalyst 12-HPW@MeSi, which was obtained by the immobilization of 12-phosphotungsticacid on a mesoporous silica framework. High-performance liquid chromatography coupled with massspectrometry was used to identify the transformation products. Results: A total of 18 transformation products were obtained and identified. Methanol was found to be involved inthe formation of 8 products formed by the addition of methanol molecules to the C-24 (25), C-20 (21) or C-20(22) double bonds of the aglycone. The transformation pathways of ginsenoside Rb1 involved deglycosylation,addition, elimination, cycloaddition, and epimerization reactions. These pathways could be elucidated in termsof the stability of the generated carbenium ion. In addition, 12-HPW@MeSi was able to maintain a 60.5%conversion rate of Rb1 after 5 cycles. Conclusion: Tandem and high-resolution mass spectrometry analysis allowed rapid and accurate identification ofthe transformation products through the characteristic fragment ions and neutral loss. Rare ginsenosides withmethoxyl groups grafted at the C-25 and C-20 positions were obtained for the first time by chemical transformationusing the composite catalyst 12-HPW@MeSi, which also enabled cyclic heterogeneous transformationand facile centrifugal separation of ginsenosides. This work provides an efficient and recyclable strategy for thepreparation of rare ginsenosides with the involvement of organic molecules.

Differentiation and identification of ginsenoside structural isomers by two-dimensional mass spectrometry combined with statistical analysis

Yang Xiu,Huanxi Zhao,Xiuli Sun,Xue Li,Shuying Liu 고려인삼학회 2019 Journal of Ginseng Research Vol.43 No.3

Background: In the current phytochemical research on ginseng, the differentiation and structural identificationof ginsenosides isomers remain challenging. In this paper, a two-dimensional mass spectrometry(2D-MS) method was developed and combined with statistical analysis for the directdifferentiation, identification, and relative quantification of protopanaxadiol (PPD)-type ginsenosideisomers. Methods: Collision-induced dissociation was performed at successive collision energy values to producedistinct profiles of the intensity fraction (IF) and ratio of intensity (RI) of the fragment ions. To amplifythe differences in tandem mass spectra between isomers, IF and RI were plotted against collision energy. The resulting data distributions were then used to obtain the parameters of the fitted curves, which wereused to evaluate the statistical significance of the differences between these distributions via the unpairedt test. Results: A triplet and two pairs of PPD-type ginsenoside isomers were differentiated and identified bytheir distinct IF and RI distributions. In addition, the fragmentation preference of PPD-type ginsenosideswas determined on the basis of the activation energy. The developed 2D-MS method was also extendedto quantitatively determine the molar composition of ginsenoside isomers in mixtures of biotransformationproducts. Conclusion: In comparison with conventional mass spectrometry methods, 2D-MS provides more directinsights into the subtle structural differences between isomers and can be used as an alternativeapproach for the differentiation of isomeric ginsenosides and natural products.

Differentiation and identification of ginsenoside structural isomers by two-dimensional mass spectrometry combined with statistical analysis

Xiu, Yang,Ma, Li,Zhao, Huanxi,Sun, Xiuli,Li, Xue,Liu, Shuying The Korean Society of Ginseng 2019 Journal of Ginseng Research Vol.43 No.3

Background: In the current phytochemical research on ginseng, the differentiation and structural identification of ginsenosides isomers remain challenging. In this paper, a two-dimensional mass spectrometry (2D-MS) method was developed and combined with statistical analysis for the direct differentiation, identification, and relative quantification of protopanaxadiol (PPD)-type ginsenoside isomers. Methods: Collision-induced dissociation was performed at successive collision energy values to produce distinct profiles of the intensity fraction (IF) and ratio of intensity (RI) of the fragment ions. To amplify the differences in tandem mass spectra between isomers, IF and RI were plotted against collision energy. The resulting data distributions were then used to obtain the parameters of the fitted curves, which were used to evaluate the statistical significance of the differences between these distributions via the unpaired t test. Results: A triplet and two pairs of PPD-type ginsenoside isomers were differentiated and identified by their distinct IF and RI distributions. In addition, the fragmentation preference of PPD-type ginsenosides was determined on the basis of the activation energy. The developed 2D-MS method was also extended to quantitatively determine the molar composition of ginsenoside isomers in mixtures of biotransformation products. Conclusion: In comparison with conventional mass spectrometry methods, 2D-MS provides more direct insights into the subtle structural differences between isomers and can be used as an alternative approach for the differentiation of isomeric ginsenosides and natural products.

A comparative study on chemical composition of total saponins extracted from fermented and white ginseng under the effect of macrophage phagocytotic function

Dan Xiao,Yang Xiu,Hao Yue,Xiuli Sun,Huanxi Zhao,Shuying Liu 고려인삼학회 2017 Journal of Ginseng Research Vol.41 No.3

In this study, white ginseng was used as the raw material, which was fermented with Paecilomyces hepiali through solid culture medium, to produce ginsenosides with modified chemical composition. The characteristic chemical markers of the products thus produced were investigated using rapid resolution liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (RRLCeQTOFeMS). Chemical profiling data were obtained, which were then subjected to multivariate statistical analysis for the systematic comparison of active ingredients in white ginseng and fermented ginseng to understand the beneficial properties of ginsenoside metabolites. In addition, the effects of these components on biological activity were investigated to understand the improvements in the phagocytic function of macrophages in zebrafish. According to the established RRLCeQTOFeMS chemical profiling, the contents in ginsenosides of high molecular weight, especially malonylated protopanaxadiol ginsenosides, were slightly reduced due to the fermentation, which were hydrolyzed into rare and minor ginsenosides. Moreover, the facilitation of macrophage phagocytic function in zebrafish following treatment with different ginseng extracts confirmed that the fermented ginseng is superior to white ginseng. Our results prove that there is a profound change in chemical constituents of ginsenosides during the fermentation process, which has a significant effect on the biological activity of these compounds.

Simultaneous determination and difference evaluation of 14 ginsenosides in Panax ginseng roots cultivated in different areas and ages by high-performance liquid chromatography coupled with triple quadrupole mass spectrometer in the multiple reaction-moni

Xiu, Yang,Li, Xue,Sun, Xiuli,Xiao, Dan,Miao, Rui,Zhao, Huanxi,Liu, Shuying The Korean Society of Ginseng 2019 Journal of Ginseng Research Vol.43 No.4

Background: Ginsenosides are not only the principal bioactive components but also the important indexes to the quality assessment of Panax ginseng Meyer. Their contents in cultivated ginseng vary with the growth environment and age. The present study aimed at evaluating the significant difference between 36 cultivated ginseng of different cultivation areas and ages based on the simultaneously determined contents of 14 ginsenosides. Methods: A high-performance liquid chromatography (HPLC) coupled with triple quadrupole mass spectrometer (MS) method was developed and used in the multiple reaction-monitoring (MRM) mode (HPLC-MRM/MS) for the quantitative analysis of ginsenosides. Multivariate statistical analysis, such as principal component analysis and partial least squares-discriminant analysis, was applied to discriminate ginseng samples of various cultivation areas and ages and to discover the differentially accumulated ginsenoside markers. Results: The developed HPLC-MRM/MS method was validated to be precise, accurate, stable, sensitive, and repeatable for the simultaneous determination of 14 ginsenosides. It was found that the 3- and 5-yr-old ginseng samples were differentiated distinctly by all means of multivariate statistical analysis, whereas the 4-yr-old samples exhibited similarity to either 3- or 5-yr-old samples in the contents of ginsenosides. Among the 14 detected ginsenosides, Rg1, Rb1, Rb2, Rc, 20(S)-Rf, 20(S)-Rh1, and Rb3 were identified as potential markers for the differentiation of cultivation ages. In addition, the 5-yr-old samples were able to be classified in cultivation area based on the contents of ginsenosides, whereas the 3- and 4-yr-old samples showed little differences in cultivation area. Conclusion: This study demonstrated that the HPLC-MRM/MS method combined with multivariate statistical analysis provides deep insight into the accumulation characteristics of ginsenosides and could be used to differentiate ginseng that are cultivated in different areas and ages.

Simultaneous determination and difference evaluation of 14 ginsenosides in Panax ginseng roots cultivated in different areas and ages by high-performance liquid chromatography coupled with triple quadrupole mass spectrometer in the multiple reaction-monitoring mode combined with multivariate statistical analysis

Yang Xiu,Xue Li,Xiuli Sun,Dan Xiao,Rui Miao,Huanxi Zhao,Shuying Liu 고려인삼학회 2019 Journal of Ginseng Research Vol.43 No.4

Background: Ginsenosides are not only the principal bioactive components but also the important indexes to the quality assessment of Panax ginseng Meyer. Their contents in cultivated ginseng vary with the growth environment and age. The present study aimed at evaluating the significant difference between 36 cultivated ginseng of different cultivation areas and ages based on the simultaneously determined contents of 14 ginsenosides. Methods: A high-performance liquid chromatography (HPLC) coupled with triple quadrupole mass spectrometer (MS) method was developed and used in the multiple reaction-monitoring (MRM) mode (HPLC-MRM/MS) for the quantitative analysis of ginsenosides. Multivariate statistical analysis, such as principal component analysis and partial least squares-discriminant analysis, was applied to discriminate ginseng samples of various cultivation areas and ages and to discover the differentially accumulated ginsenoside markers. Results: The developed HPLC-MRM/MS method was validated to be precise, accurate, stable, sensitive, and repeatable for the simultaneous determination of 14 ginsenosides. It was found that the 3- and 5-yrold ginseng samples were differentiated distinctly by all means of multivariate statistical analysis, whereas the 4-yr-old samples exhibited similarity to either 3- or 5-yr-old samples in the contents of ginsenosides. Among the 14 detected ginsenosides, Rg1, Rb1, Rb2, Rc, 20(S)-Rf, 20(S)-Rh1, and Rb3 were identified as potential markers for the differentiation of cultivation ages. In addition, the 5-yr-old samples were able to be classified in cultivation area based on the contents of ginsenosides, whereas the 3- and 4-yr-old samples showed little differences in cultivation area. Conclusion: This study demonstrated that the HPLC-MRM/MS method combined with multivariate statistical analysis provides deep insight into the accumulation characteristics of ginsenosides and could be used to differentiate ginseng that are cultivated in different areas and ages.

A comparative study on chemical composition of total saponins extracted from fermented and white ginseng under the effect of macrophage phagocytotic function

Xiao, Dan,Xiu, Yang,Yue, Hao,Sun, Xiuli,Zhao, Huanxi,Liu, Shuying The Korean Society of Ginseng 2017 Journal of Ginseng Research Vol.41 No.3

In this study, white ginseng was used as the raw material, which was fermented with Paecilomyces hepiali through solid culture medium, to produce ginsenosides with modified chemical composition. The characteristic chemical markers of the products thus produced were investigated using rapid resolution liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (RRLC-QTOF-MS). Chemical profiling data were obtained, which were then subjected to multivariate statistical analysis for the systematic comparison of active ingredients in white ginseng and fermented ginseng to understand the beneficial properties of ginsenoside metabolites. In addition, the effects of these components on biological activity were investigated to understand the improvements in the phagocytic function of macrophages in zebrafish. According to the established RRLC-QTOF-MS chemical profiling, the contents in ginsenosides of high molecular weight, especially malonylated protopanaxadiol ginsenosides, were slightly reduced due to the fermentation, which were hydrolyzed into rare and minor ginsenosides. Moreover, the facilitation of macrophage phagocytic function in zebrafish following treatment with different ginseng extracts confirmed that the fermented ginseng is superior to white ginseng. Our results prove that there is a profound change in chemical constituents of ginsenosides during the fermentation process, which has a significant effect on the biological activity of these compounds.