Study On The Metabolite In Vivo Of Chloroform Extracts And N-butanol Extracts Chemical Composition From Clerodendranthus Spicatus

The whole plant of Clerodendranthus spicatus（Thunb.）C.Y.Wu.has been used as a folk medicine among Dai people for ages.It is for many disease of urine system,such as lithangiuria,acute and chronic glomerulonephritis etc..Previous research in our lab showed that both chloroform extract and n-butanol extract could significantly reduce the level of Heymann rat albuminuria and made the rat recover from nephropathy.Which compounds can be absorbed and distributed into kidney is uncertain although systemic chemical investigation of the two extract were finished.So,the main purpose of the study is to uncover the ingredients absorbed and distributed into targeted kidney so as to give evidence for pharmacological substance determination.Ten polymethoxy-flavones（PMFs）and three phenolic acids（PAs）of C.spicatus were subjected to UPLC-Q-TOF-MS/MS analysis to obtain the characteric ions and possible cheverage pathways which could be used for structure identification of minor ingredients in biological samples.Cleverages at C-ring of PMFs are reported including retro Diels-Alder（RDA）in the dissertation.PMFs with two methoxy orth-substituted in A or B ring show more frequently the fragmentation ions with neutral loss of CH₄,C₂H₆ while lower abundance of ^1,3A⁺,^1,3B⁺,^0,2B⁺through direct C-ring cleverage which can be the characteristic of these compounds.Characteristic fragmentation ions for phenolic acids have been obtain,such as C₈H₅O₃（m/z 149.0341）,C₇H₅O₂（m/z 121.0367）for compound7,8-dihydroxy-3-methyl-isochromanone are reported for the first time.UPLC-DAD、UPLC-QTOF-MS/MS were then used for analysis of biological specimen,such as rat plasma,kidney homogenate and urine after ig chloroform extract and n-butanlo extract of C.spicatus to investigate the PMFs,PAs absorbed and their related metabolites.The results for ig chloroform extract show that two PMFs were detected in rat plasma named 5,6,7,3′,4′-pentamethoxy-flavone,5-hydroxy-6,7,4′-trimethoxy-flavone which were also found in kidney with additional one named 5,6,7,4′-tetramethoxy-flavone.The last one was then deduced to be a metabolite.6,7,4′-trimethoxy-flavone-5-O-glucuronic acid was detected in urine.The result for ig n-butanol extract showed that caffeic acid and rosmarinic acid were detected in both rat plasma and kidney while rosmarinic acid and caffeic acid glucuronidate were found in urine.Cffeic acid metabolites with sulfonation,methylation were also detected in rat plasma and kidney.Above results suggest that the pharmacological substance of C.spicatus will be focused on the four ingredients named5,6,7,3′,4′-pentamethoxy-flavone,5-hydroxy-6,7,4′-trimethoxy-flavone,caffeic acid and rosmarinic acid.