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
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
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.