Novel Analysis Procedure for Red Ginseng Polysaccharides by Matrix-assisted Laser Desorption/ionization Time-of-flight/time-of-flight Mass Spectrometry

Overview

Journal J Ginseng Res

Date 2021 Nov 22

PMID 34803423

Citations 2

Authors

Ye Rin Jin

Myung Jin Oh

Heung Joo Yuk

Hyun Joo An

Dong Seon Kim

Affiliations

Soon will be listed here.

Abstract

Background: Red ginseng polysaccharides (RGPs) have been acknowledged for their outstanding immunomodulation and anti-tumor activities. However, their studies are still limited by the complexity of their structural features, the absence of purification and enrichment methods, and the rarity of the analytical instruments that apply to the analysis of such macromolecules. Thus, this study is an attempt to establish a new mass spectrometry (MS)-based analysis procedure for RGPs.

Methods: Saponin pre-excluded powder of RG (RG-SPEP, 10 mg) was treated with 200 μL of distilled water and centrifuged for 5 h at 1000 rpm and 85 °C. Ethanol-based precipitation and centrifugation were applied to obtain RGPs from the heated extracts. Further, endo-carbohydrase treatments were performed to produce specific saccharide fragments. Solid-phase extraction (SPE) processes were implemented to purify and enrich the enzyme-treated RGPs, while matrix-assisted laser desorption/ionization time-of-flight/time-of-flight (MALDI-TOF/TOF) MS was employed for the partial structural analysis of the obtained RGPs.

Results: Utilizing cellulase, porous graphitized carbon (PGC), hydrophilic interaction chromatography (HILIC), and MALDI-TOF/TOF MS, the neutral and acidic RGPs were qualitatively analyzed. Hex and Hex 18 (cellulose analogs) were determined to be novel neutral RGPs. Additionally, the [Unknown + Hex] species were also determined as new acidic RGPs. Furthermore, HexA (H) was determined as another form of the acidic RGPs.

Conclusion: Compared to the previous methods of analysis, these unprecedented applications of HILIC-SPE and MALDI-TOF/TOF MS to analyze RGPs proved to be fairly effective for fractionating and detecting neutral and acidic components. This new procedure exhibits great potential as a specific tool for searching and determining various polysaccharides in many herbal medicines.

Citing Articles

Research Progress on Extraction, Isolation, Structural Analysis and Biological Activity of Polysaccharides from Panax Genus.

Zhang S, Ding C, Liu X, Zhao Y, Ding Q, Sun S Molecules. 2023; 28(9).

PMID: 37175143 PMC: 10179830. DOI: 10.3390/molecules28093733.

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry combined with chemometrics to identify the origin of Chinese medicinal materials.

Fang H, Chen Y, Wu H, Chen Y, Wang T, Yang J RSC Adv. 2022; 12(26):16886-16892.

PMID: 35754890 PMC: 9171747. DOI: 10.1039/d2ra02040h.

References

Clark A, Kaleta E, Arora A, Wolk D . Matrix-assisted laser desorption ionization-time of flight mass spectrometry: a fundamental shift in the routine practice of clinical microbiology. Clin Microbiol Rev. 2013; 26(3):547-603. PMC: 3719498. DOI: 10.1128/CMR.00072-12. View

Jensen P, Mysling S, Hojrup P, Jensen O . Glycopeptide enrichment for MALDI-TOF mass spectrometry analysis by hydrophilic interaction liquid chromatography solid phase extraction (HILIC SPE). Methods Mol Biol. 2013; 951:131-44. DOI: 10.1007/978-1-62703-146-2_10. View

Guo N, Bai Z, Jia W, Sun J, Wang W, Chen S . Quantitative Analysis of Polysaccharide Composition in by HPLC-ESI-TOF-MS. Molecules. 2019; 24(14). PMC: 6681038. DOI: 10.3390/molecules24142526. View

Seo Y, Oh M, Park J, Ko J, Kim J, An H . Comprehensive Characterization of Biotherapeutics by Selective Capturing of Highly Acidic Glycans Using Stepwise PGC-SPE and LC/MS/MS. Anal Chem. 2019; 91(9):6064-6071. DOI: 10.1021/acs.analchem.9b00603. View

Wu W, Jiao C, Li H, Ma Y, Jiao L, Liu S . LC-MS based metabolic and metabonomic studies of Panax ginseng. Phytochem Anal. 2018; 29(4):331-340. DOI: 10.1002/pca.2752. View

Cheong K, Wu D, Zhao J, Li S . A rapid and accurate method for the quantitative estimation of natural polysaccharides and their fractions using high performance size exclusion chromatography coupled with multi-angle laser light scattering and refractive index detector. J Chromatogr A. 2015; 1400:98-106. DOI: 10.1016/j.chroma.2015.04.054. View

Hyun S, Kim S, Seo H, Youn S, Kyung J, Lee Y . Physiological and pharmacological features of the non-saponin components in Korean Red Ginseng. J Ginseng Res. 2020; 44(4):527-537. PMC: 7322739. DOI: 10.1016/j.jgr.2020.01.005. View

Amicucci M, Galermo A, Guerrero A, Treves G, Nandita E, Kailemia M . Strategy for Structural Elucidation of Polysaccharides: Elucidation of a Maize Mucilage that Harbors Diazotrophic Bacteria. Anal Chem. 2019; 91(11):7254-7265. DOI: 10.1021/acs.analchem.9b00789. View

Ji L, Jie Z, Ying X, Yue Q, Zhou Y, Sun L . Structural characterization of alkali-soluble polysaccharides from C. A. Meyer. R Soc Open Sci. 2018; 5(3):171644. PMC: 5882694. DOI: 10.1098/rsos.171644. View

10.

Leung K, Wong A . Pharmacology of ginsenosides: a literature review. Chin Med. 2010; 5:20. PMC: 2893180. DOI: 10.1186/1749-8546-5-20. View

11.

Melmer M, Stangler T, Premstaller A, Lindner W . Comparison of hydrophilic-interaction, reversed-phase and porous graphitic carbon chromatography for glycan analysis. J Chromatogr A. 2010; 1218(1):118-23. DOI: 10.1016/j.chroma.2010.10.122. View

12.

Ozcan S, Barkauskas D, Ruhaak L, Torres J, Cooke C, An H . Serum glycan signatures of gastric cancer. Cancer Prev Res (Phila). 2013; 7(2):226-35. PMC: 3946197. DOI: 10.1158/1940-6207.CAPR-13-0235. View

13.

Kailemia M, Ruhaak L, Lebrilla C, Jonathan Amster I . Oligosaccharide analysis by mass spectrometry: a review of recent developments. Anal Chem. 2013; 86(1):196-212. PMC: 3924431. DOI: 10.1021/ac403969n. View

14.

Xu J, Yue R, Liu J, Ho H, Yi T, Chen H . Structural diversity requires individual optimization of ethanol concentration in polysaccharide precipitation. Int J Biol Macromol. 2014; 67:205-9. DOI: 10.1016/j.ijbiomac.2014.03.036. View

15.

Lee S, In G, Han S, Lee M, Lee J, Shin K . Structural characteristics of a red ginseng acidic polysaccharide rhamnogalacturonan I with immunostimulating activity from red ginseng. J Ginseng Res. 2020; 44(4):570-579. PMC: 7322754. DOI: 10.1016/j.jgr.2019.05.002. View

16.

Hung W, Wang S, Chen Y, Yu H, Chen C, Yang W . MALDI-TOF MS analysis of native and permethylated or benzimidazole-derivatized polysaccharides. Molecules. 2012; 17(5):4950-61. PMC: 6268202. DOI: 10.3390/molecules17054950. View

17.

Caffall K, Mohnen D . The structure, function, and biosynthesis of plant cell wall pectic polysaccharides. Carbohydr Res. 2009; 344(14):1879-900. DOI: 10.1016/j.carres.2009.05.021. View

18.

Sun X, Matsumoto T, Yamada H . Purification of an anti-ulcer polysaccharide from the leaves of Panax ginseng. Planta Med. 1992; 58(5):445-8. DOI: 10.1055/s-2006-961510. View

19.

Baek S, Bae O, Park J . Recent methodology in ginseng analysis. J Ginseng Res. 2013; 36(2):119-34. PMC: 3659581. DOI: 10.5142/jgr.2012.36.2.119. View

20.

Jiao L, Wan D, Zhang X, Li B, Zhao H, Liu S . Characterization and immunostimulating effects on murine peritoneal macrophages of oligosaccharide isolated from Panax ginseng C.A. Meyer. J Ethnopharmacol. 2012; 144(3):490-6. DOI: 10.1016/j.jep.2012.09.004. View