Studies On The Chemical Constituents From The Bulbus Fritillariae Cirrhosae And Quality Assessment Of Fritillaria Species

The genus Fritillaria belongs to family Liliaceae,which includes about 130 species worldwide and occurs mainly in the temperate regions of the northern hemisphere.Most species are distributed in Cyprus,Iran and Turkey.The bulbs of many Fritillaria species ('Beimu' in Chinese) have been used as antitussive,anti-asthmatic and expectorant agents in traditional Chinese medicine for more than 2000 years.Fritillaria cirrhosa D. Don and Fritillaria delavayi Franch.are recorded as two of plant sources of 'Chuanbeimu' in China Pharmacopoeia(2005 Edition),which documented to treat cough, bronchitis and hemoptysis.In order to search for biologically active constituents,as well as develop a method for the quality control of Fritillaria raw materials,the bulbs of F. cirrhosa D.Don and F.delavayi Franch.were subjected to systematically phytochemical studies.The analytical methods based on HPLC-ELSD and HPLC-DAD technologies were also established.Thirty-one compounds were isolated from the methanol extracts of the bulbs of F. cirrhosa by various column chromatographies.On the basis of physicochemical and spectral analysis(including IR,UV,HRESI-MS,ESI-MS,¹H NMR,¹³C NMR,DEPT, ¹H-¹H COSY,HMQC,HMBC,NOESY and X-ray),structures of twenty-seven compounds were determined,as demissidine(QB-3),chuanbeinone(QB-9),delavinone (QB-10),verticinone(QB-11),imperialine(QB-12),puqienine B(QB-16),isodelavine (QB-23),isoforticine(QB-24),petilidine(QB-25),verticine(QB-26),peimisine (QB-27),cirrhosinine A(QB-35),cirrhosinine B(QB-46),demissidine-3-O-β-D-gluco-pyranosyl (1→4) glucopyranoside(QB-65),solanidine 3-O-α-L-rhamnopyranosyl- (1→2)-[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranoside(QB-75),E-cinnamic acid (QB-P-3),2-monopalmitin(QB-P-5),uracil(QB-N-2),thymine(QB-N-3),cytidine (QB-N-6),inosine(QB-N-7),uridine(QB-N-8),guanosine(QB-N-10),thymidine (QB-N-11),adenosine(QB-N-12),β-sitosterol(QB-P-1),daucosterol(QB-P-6).Among them,there are five new compounds,including QB-23,QB-24,QB-35,QB-46 and QB-65;eight compounds,QB-P-3,QB-P-5,QB-N-2,QB-N-3,QB-N-6,QB-N-7, QB-N-8 and QB-N-10 were isolated from Fritillaria genus for the first time;QB-9, QB-10,QB-12,QB-16,QB-25,QB-75,QB-P-1 and QB-P-6 were isolated from this plant for the first time.Thirty-three compounds were isolated from the bulbs of F.delavayi,and twenty-nine of them were identified as chuanbeinone(LBM-11),delavinone(LBM-12), delavidine(LBM-14),ebeiedinone(LBM-15),verticinone(LBM-16),imperialine (LBM-17),isodelavine(LBM-31),delavine(LBM-41),ebeiedine(LBM-42), isoverticine(LBM-46),verticine(LBM-48),peimisine(LBM-50), E-3,4,5-trimethoxycinnamie acid(BM-2),E-p-methoxycinnamie acid(BM-3), E-cinnamic acid(BM-4),E-p-hydroxycinnamic acid(BM-7),E-p-hydroxycinnamic acid methyl ester(BM-8),ferulie acid(BM-11),caffeic acid(BM-12),1-O-feruloylglycerol (BM-15),uracil(LB-1),thymine(LB-2),uridine(LB-5),guanosine(LB-7),thymidine (LB-8),adenosine(LB-9),β-sitosterol(BM-1),daucosterol(BM-17),atractylenolideâ…¢(BM-5).Among them,LBM-14 is a new compound;eight compounds,BM-2,BM-3, BM-5,BM-7,BM-8,BM-11,BM-12 and BM-15 were obtained from this genus for the first time,and sixteen compounds LBM-15,LBM-16,LBM-31,LBM-42,LBM-46, LBM-48,LBM-50,BM-1,BM-4,BM-17,LB-1,LB-2,LB-5,LB-7,LB-8 and LB-9 were isolated from this plant for the first time.A facile method based on high performance liquid chromatography coupled with evaporative light scattering detector(HPLC-ELSD) was developed for the simultaneous quantitative determination of eight alkaloids including pingbeimine A,peimisine, verticine,verticinone,isodelavine,demissidine-3-O-β-D-glucopyranosyl(1→4)-glucopyranoside in 60 batches of samples from 13 Fritillaria species.The chromatographic patterns of 13 Fritillaria species were also established,and the chemotaxonomic characteristics were proposed.Furthermore,a simple method based on high performance liquid chromatography coupled with diode-array detector(HPLC-DAD) was developed for the simultaneous determination of nine nucleosides and bases, including uracil,cytidine,guanine,uridine,thymine,inosine,guanosine,thymidine, adenosine in the same above 60 batches of samples.The results showed that uridine, guanosine and adenosine are the major nucleosides in Fritillaria bulbs,and may be the main bioactive constituents other than alkaloids.To conclude,The newly developed HPLC-ELSD and HPLC-DAD methods for simultaneous determination of eight alkaloids and nine nucleosides could be used as powerful tools for quality control of Fritillaria bulbs.In addition,advance in the research of chemical constituents,pharmacological activities and quality control of medicinal plants from Fritillaria genus were reviewed with 223 references.