Methodology Research On Metabolism Of Huan-Nao-Yi-Cong-Fang And Flavonoid Components From Invasive Weeds By LC-IT-TOF-MS

Huan-Nao-Yi-Cong-Fang (HNYCF) is one potential prescription in treating Alzheimer's disease. This study explored the metabolites of the seven bioactive components of HNYCF in rat plasma to explain the metabolite patterns of HNYCF and give useful data for further studying action mechanism of HNYCF. The other study in this article is the identification of flavonoid components from invasive weeds. This article aimed to develop a useful method to identify nontarget flavonoid components from invasive weeds in methodology by hybrid ion trap/time-of-flight mass spectrometry coupled with high-performance liquid chromatography (LC-IT-TOF-MS). Compared with simple controlling methods for dangerous exotic weeds, utilization of these plants might give more benefits. 1. The following seven active components have been used as quality chemical markers of HNYCF due to their biological significance and high contents in crude plant materials:ferulic acid,2,3,4,5'-tetrahydroxystilbene-2-O-?-D-glucoside,berberine hydrochloride, emodin, ginsenoside Rgl, ginsenoside Re and ginsenoside Rbl. Simultaneously identification of the metabolites the seven bioactive components of HNYCF in rat plasma using LC-IT-TOF-MS combined with MetID solution 1.0 software. Using a gradient elution consisted of mobile phase A (0.5% formic acid) and mobile phase B (methanol). A binary gradient elution was performed:0-23.6 min, a linear gradient from 46% to 100% B at a flow rate of 0.5 mL/min. The sample injection volume was 5 ?L. the collision energy was set at 50% for MS2 and 30% for MS3, respectively. The detector voltage and the ion accumulation time were set at 1.70 kV and 30 ms, respectively. A total of 10 metabolites were found from the rat plasma 2-hour after oral administration of HNYCF dosage, of which 2 metabolites of emodin 7-hydroxyemodin and 5,7-dihydroxyemodin were observed for the first time.2. This study has developed a novel diagnostic fragment-ion method to rapidly screen and identify nontarget flavonoid components from invasive weeds by hybrid ion trap/time-of-flight mass spectrometry coupled with high-performance liquid chromatography. This method consists of the following three steps:first, the diagnostic fragment ion (DFI) for every chemical family was proposed; second, the flavonoid components were screened out based on the extracted ion chromatograms (EIC) of the DFIs; finally, the structures of the components screened out were proposed via manual fragment comparison._Using a gradient elution consisted of mobile phase A (0.5% formic acid) and mobile phase B (methanol). A binary gradient elution was performed:0-40 min, a linear gradient from 35% to 67%B at a flow rate of 0.5 mL/min. The sample injection volume was 5 ?L. the collision energy was set at 40% for MS2 and 20% for MS3, respectively. The detector voltage and the ion accumulation time were set at 1.70 kV and 30 ms, respectively. The diagnostic fragment-ion method successfully applied to the detection flavonoid components in three invasive weeds Flaveria bidentis (L.) Kuntze, Mikania micrantha Kunth and Solanum rostratum Dunal.6The results strongly demonstrate that the diagnostic fragment-ion method is highly useful for the flavonoid identification of invasive weeds.