Isolation，Purification, Structure Elucidation Of Triterpenoids From Surface Layer Of Poria Cocos And Study On Fingerprints Of Traditional Chinese Medicine Poria Cocos (Schw.) Wolf

Poria cocos（P.cocos）, the dry sclerotia of Poriacocos（Schw.）Wolf which is a kind of mycetes belonging toPolyPoraceae,is a valuable traditional Chinese medicine witha more than thousands of years using history. It is oftencombined in many traditional Chinese prescriptions as adiuretic,sedative, and tonic. Modern pharmacology researcheson Poria cocos show that the triterpenoid is one of its maineffective component. The surface layer of Poria cocos（Fu-Ling-Pi in Chinese） as the epidermis of the sclerotia wereusually abondoned during processing.However,through recentyears researches, we found that the surface layer of Poria cocosnot only contain many kinds of compositions of triterpenoid,butsome of them were particular. In this paper，the extraction，isolation，purification of triperpenes in the surface layer ofPoria cocos were studied and the UPLC-UV-MS fingerprints oftriterpenoid constituents of Poria cocos in differentmedicinal parts and from different sources were set up.In this study,we adopt alkali-acid treatment to gettriterpenoid. The surface layer of Poria cocos was extracted with MeOH under reflux.The MeOH solution was concentrated toan extract,which was mixed with H₂O and extracted with Et2O.TheEt₂O-soluble fraction was further extracted with saturatedNaHCO₃aqueous solution and then5%NaOH aqueous solution.Bothof them wererespectively acidize with6M HCl and then extractedwith Et2O. Finally,we obtained two fractions: FractionA（saturated NaHCO₃aqueous-soluble fraction） and FractionB（5%NaOH-soluble fraction）.By using modern analysis techniques such as silica gel，thin-Layer chromatography， preparative chromatography andnuclear magnetic resonance.etc, Fraction A（saturated NaHCO₃aqueous-soluble fraction） was obtained eight compounds,andfive of them had their structures been elucidated from¹H-NMRand¹³C-NMR spectral data.These were as follows: Poricoic acidA、Poricoic acid B、Poricoic acid AM、Poricoic acid G、Dehydrotumulosic acid. The structures of another threecompounds have not yet been elucidated,but their purity andmolecular weight had been confirmed by UPLC-UV-MS. Thisresearch will be useful for the further study of effectivematerial basic of Poria cocos, especially the characteristiccompositions of3,4-seco-lanostan in the surface layer of Poriacocos.To develop the Ultra performance liquid chromatography-ultra violet-mass spectrometry （UPLC-UV-MS） fingerprint of thePoria cocos（Schw.）Wolf. Twenty-two peaks were found indifferent sources of Poria cocos,and nine peaks of them werecommon and rich.With dehydrotumulosic acid、poricoic acid A、poricoic acid B、 poricoic acid AM、 pachymic acid anddehtdrapachymic acid as reference substance, meanwhile, comparaing the m/z of ion peaks with relevant literatures,seventeen peaks were identified preliminarily.The results ofmethod validation met technical standard of fingerprints;Through the similarity and cluster analysis, we concluded thattriterpenoid constituents in Poria peel and Poria piece werevery different. The quality of Poria cocos is relation to itsproducing area.This method is stable and reliable with a goodreproducibility and can provide a reference standard for thequality control of Poria cocos.