Generic placeholder image

The Natural Products Journal

Editor-in-Chief

ISSN (Print): 2210-3155
ISSN (Online): 2210-3163

Research Article

One-step Separation and Purification of Four Phenolic Acids from Stenoloma chusanum (L.) Ching by Medium-pressure Liquid Chromatography and High-speed Counter-current Chromatography

Author(s): Tianyun Li, Xiling Dai, Yichen Li, Guozheng Huang* and Jianguo Cao*

Volume 9, Issue 2, 2019

Page: [138 - 143] Pages: 6

DOI: 10.2174/2210315508666181004115422

Price: $65

Abstract

Background: Stenoloma chusanum (L.) Ching is a Chinese traditional medicinal fern with high total flavonoid and total phenolic content. Traditionally, phenolic compounds were separated by using column chromatography, which is relatively inefficient.

Objective: This study aims to use an efficient method to separate natural products from S. chusanum by Medium-Pressure Liquid Chromatography (MPLC) and High-Speed Counter-Current Chromatography (HSCCC).

Methods: In the present research, firstly, a sample (2.5 g) from the dichloromethane extract of S. chusanum was separated by MPLC. Next, fraction P5 was purified by HSCCC with a two-phase solvent system composed of hexane-ethyl acetate-methanol-water (HEMWat) at a volume ratio of 2:4:1:4 (v/v/v/v).

Result: Four phenolic acids were obtained and their structures were identified by means of NMR and ESI-mass analysis. They were identified as: 1) protocatechuic acid (34 mg, purity 90.1%), 2) syringic acid (66 mg, purity 99.0%), 3) p-hydroxybenzoic acid (5 mg, purity 91.2%) and 4) vanillic acid (6 mg, purity 99.3%).

Conclusion: The combination of MPLC and HSCCC is a high-efficiency separation method for natural products. This is the first report with regard to the separation of four phenolic acids in one step by MPLC and HSCCC from S. chusanum (L.) Ching.

Keywords: Fern, Stenoloma chusanum, HSCCC, MPLC, phenolic acid, vanillic acid.

Graphical Abstract
[1]
Wu, S.; Li, J.; Wang, Q.; Cao, H.; Cao, J.; Xiao, J. Seasonal dynamics of the phytochemical constituents and bioactivities of extracts from Stenoloma chusanum (L.) Ching. Food Chem. Toxicol., 2017, 108, 458-466.
[2]
Luo, Y.; Xiao, X.; Wang, Z. Extraction, separation, purification and structural analysis of polysaccharide from Stenoloma chusanum (L.) Ching. Chem. Ind. Forest Prod., 2009, 29(1), 68-72.
[3]
Li, M.; Luo, Y.; Li, H. Studies on chemical constituents of Stenoloma chusana (L.) Ching flavonoids. J. Sichuan Norm. Univ., 2009, 32(3), 358-360.
[4]
Ren, B.; Xia, B.; Li, W.L.; Wu, J.; Zhang, H. Two novel phenolic compounds from Stenoloma chusanum and their antifungal activity. Chem. Nat. Compd., 2009, 45(2), 182-186.
[5]
Stalikas, C.D. Extraction, separation, and detection methods for phenolic acids and flavonoids. J. Sep. Sci., 2007, 30(18), 3268-3295.
[6]
Liang, Z.; Li, B.; Liang, Y.; Su, Y.; Ito, Y. Separation and purification of two minor compounds from Radix isatidis by integrative MPLC and HSCCC with preparative HPLC. J. Liq. Chromatogr. Relat. Technol., 2015, 38(5), 647-653.
[7]
Wang, X.; Liang, Y.; Zhu, L.; Xie, H.; Li, H.; He, J.; Pan, M.; Zhang, T.; Ito, Y. Preparative isolation and purification of flavone C-glycosides from the leaves of Ficus microcarpa Lf by medium-pressure liquid chromatography, high-speed countercurrent chromatography, and preparative liquid chromatography. J. Liq. Chromatogr. Relat. Technol., 2010, 33(4), 462-480.
[8]
Ito, Y. Golden rules and pitfalls in selecting optimum conditions for high-speed counter-current chromatography. J. Chromatogr. A, 2005, 1065(2), 145-168.
[9]
Kai, X.; Lü, H.; Qü, B.; Hu, S.; Song, J. High-speed counter-current chromatography preparative separation and purification of phloretin from apple tree bark. Separ. Purif. Tech., 2010, 72(3), 406-409.
[10]
Li, F.; Lin, Y.; Wang, X.; Geng, Y.; Wang, D. Preparative isolation and purification of capsaicinoids from Capsicum frutescens using high-speed counter-current chromatography. Separ. Purif. Tech., 2009, 64(3), 304-308.
[11]
Sutherland, I.A.; Fisher, D. Role of counter-current chromatography in the modernisation of Chinese herbal medicines. J. Chromatogr. A, 2009, 1216(4), 740-753.
[12]
Sutherland, I.; Hawes, D.; Ignatova, S.; Janaway, L.; Wood, P. Review of progress toward the industrial scale up of CCC. J. Liq. Chromatogr. Relat. Technol., 2005, 28(12-13), 1877-1891.
[13]
Hu, R.; Pan, Y. Recent trends in counter-current chromatography. Trends Analyt. Chem., 2012, 40, 15-27.
[14]
Wang, M.; Zou, H.; Chen, Q.; Cao, J.; Aisa, H.A.; Huang, G. Isolation of new polyacetylenes from the roots of Eurycoma longifolia via high-speed counter-current chromatography. J. Chromatogr. B, 2017, 1055-1056, 39-44.
[15]
Chen, T.; Li, H.; Tan, L.; Li, Y. Separation of 4′-demethyldeoxypodophyllotoxin from Sinopodophyllum emodi by medium-pressure LC and high-speed counter-current chromatography guided by HPLC-MS. Sep. Sci. Technol., 2017, 52(8), 1423-1429.
[16]
Qi, L.; Wang, C.; Yuan, C. Isolation and analysis of ginseng: Advances and challenges. Nat. Prod. Rep., 2011, 28(3), 467-495.
[17]
Liang, J.; Meng, J.; Wu, D.; Guo, M.; Wu, S. A novel 9×9 map-based solvent selection strategy for targeted counter-current chromatography isolation of natural products. J. Chromatogr. A, 2015, 1400, 27-39.
[18]
Cong, L.; Hu, Y.; Hu, Y.; Zhang, Y.; Li, S.; Liu, C. Application of high-performance counter-current chromatography and medium-pressure liquid chromatography for rapid isolation of lactones from Ligusticum chuanxiong hort. J. Liq. Chromatogr. Relat. Technol., 2014, 37(8), 1187-1198.
[19]
Wang, P.; Liu, Y.; Chen, T.; Xu, W.; You, J.; Liu, Y.; Li, Y. One-step separation and purification of three lignans and one flavonol from Sinopodophyllum emodi by medium-pressure liquid chromatography and high-speed counter-current chromatography. Phytochem. Anal., 2013, 24(6), 603-607.
[20]
Li, H.; Chen, F. Preparative isolation and purification of chuanxiongzine from the medicinal plant Ligusticum chuanxiong by high-speed counter-current chromatography. J. Chromatogr. A, 2004, 1047(2), 249-253.
[21]
Han, Q.; Wong, L.; Yang, N.; Song, J.; Qiao, C.; Yiu, H.; Ito, Y.; Xu, H. A simple method to optimize the HSCCC two-phase solvent system by predicting the partition coefficient for target compound. J. Sep. Sci., 2008, 31(6-7), 1189-1194.
[22]
Zhu, L.; Li, H.; Liang, Y.; Wang, X.; Xie, H.; Zhang, T.; Ito, Y. Application of high-speed counter-current chromatography and preparative high-performance liquid chromatography mode for rapid isolation of anthraquinones from Morinda officinalis How. Separ. Purif. Tech., 2009, 70(2), 147-152.
[23]
Yang, Y.; Gu, D.Y.; Yili, A.; Zhao, Y.X.; He, D.J.; Aisa, H.A. One-step separation and purification of rupestonic acid and chrysosptertin B from Artemisia rupestris L. by high-speed counter-current chromatography. Phytochem. Anal., 2010, 21(2), 205-209.
[24]
Itoh, A.; Isoda, K.; Kondoh, M.; Kawase, M.; Kobayashi, M.; Tamesada, M.; Yagi, K. Hepatoprotective effect of syringic acid and vanillic acid on concanavalin A-induced liver injury. Biol. Pharm. Bull., 2009, 32(7), 1215-1219.
[25]
Min, S.W.; Ryu, S.N.; Kim, D.H. Anti-inflammatory effects of black rice, cyanidin-3-O-β-D-glycoside, and its metabolites, cyanidin and protocatechuic acid. Int. Immunopharmacol., 2010, 10(8), 959-966.
[26]
Davis, B.D. Inhibition of Escherichia coli by p-aminobenzoic acid and its reversal by p-hydroxybenzoic acid. J. Exp. Med., 1951, 94(3), 243-254.
[27]
Wang, W.; Yang, C.R.; Zhang, Y.J. Phenolic constituents from the fruits of Amomum tsao-ko (Zingiberaceae). Acta Bot. Yunnanica., 2009, 31(3), 284-288.
[28]
Guo, L.; Jiang, L.; Wu, L.; Guo, W.; Zhu, Y.; Zhang, X. Studies on the chemical constituents of the stems and leaves of Acanthopanax sessiliflorus (Rupr. et Maxim.) Seem. J. Shenyang Pharm. Univ., 2002, 19(3), 180-182.
[29]
Chen, Q.; Wu, L.; Ruan, L. Chemical studies on the constituents of Lophatherum gracile Brongn. J. Shenyang Pharm. Univ., 2002, 19(4), 257-259.

Rights & Permissions Print Cite
© 2024 Bentham Science Publishers | Privacy Policy