Studies On Some Chemical Components Of Veratrum Maaekii Regell Over Ground Part And Pharmacokinetics And Metabolism Ofalkaloids

Veratrum maaekii Regel is a liliaceous?Liliaceae?hellebore?Veratrum Linn.?plants,according to“flora of China”records:Veratrum maaekii Regel is mainly distributed in the northeast,henan,shandong,Inner Mongolia etc.Veratrum roots and rhizomes of medicinal plants can be treated stroke,epilepsy,aphasia,diarrhea,headaches,psoriasis and other diseases,the main chemical ingredients are steroidal alkaloids,flavonoids,stilbenes,sterols.The chemical composition of ethanol extract of the ground part of Veratrum maackii Regel were studied systematically.By modern chromatographic methods,such as repeated silica gel column chromatography,MCI column chromatography and high-performance liquid preparation and other modern chromatographic methods,9 compounds were isolated from the ethanol extract of rhizomes of Veratrum maackii Regel overground part.By spectroscopic methods(¹H-NMR,¹³C-NMR,HMBC,HMQC,ESI-MS and UV)and they are identified as mulberrosideA?ls01?;resveratrol-4,3?-O-?-D-diglucoside?ls02?;cis-mulberrosideA?ls03?;GentifolinK?ls04?;resveratrol-3,5-O-?-D-diglucoside?ls05?;oxyresveratrol-4'-O-?-D-glucoside?ls06?;oxyresveratrol-3-O-?-D-glucoside?ls07?;oxyresveratrol?ls08?;resveratrol?ls09?.ls03,ls04 and ls05 are isolated from maackii Regel overground part for the first time.To establish a high performance capillary electrophoresis method with ultraviolet?HPLC-UV?method for the quality control of the ground part of Veratrum maackii Regel.While using a high performance capillary electrophoresis method with diode array detection?HPLC-DAD?method further determine information about the compound.The experimental determination of the content of which eight stilbenes component,and a preliminary comparison of the difference Veratrum maackii Regel content veratridine aboveground and underground portions.Comprehensive utilization of liquid chromatograph with QTRAP 4000 series triple quadrupole mass spectrometer technology to research the alkaloids Veratramine and 12?-hydroxylveratroylzy-gadenine metabolism in mice.Respectively administered to the mice urine,plasma,feces,cerebellum and cerebral cortex samples were EMS mode scanning,no different from the samples found in rats administered new metabolites.Using MRM-IDA-EPI scan mode,setting its Q1 and Q3 values of product characteristics and in vivo metabolism in rat liver microsomes in rats according to the corresponding alkaloids.It was found that veratramine had five metabolites in mouse urine,plasma,feces,cerebellum and cerebral cortex,the main metabolic pathway for the hydroxylated and sulfonated.12?-hydroxylveratroylzygadenine found eight metabolites in mouse urine,plasma,feces,cerebellum and cerebral cortex,the main metabolic pathway for the hydrolysis of ester bonds,dehydrogenation,dehydrocyclization hydrolysis of ester bonds,hydroxylation,demethylation and hydrolysis of ester bonds acetylated.Both alkaloids in cerebellum and cerebral cortex can be detected in the original drug,maybe prove alkaloids permeable blood-brain barrier into the brain tissue.To establish a rapid,accurate,sensitive and convenient method of LC-MS-MS to detect veratramine plasma concentration in mice plasma.Plasma samples were extracted protein precipitation veratramine biological sample and the internal standard jervine.Using 0.1%aqueous formic acid-acetonitrile as a mobile phase gradient,flow rate of 0.3 mL/min,with ESI positive ionization mode and multiple reaction monitoring mode?MRM?as a condition of mass spectrometry,and ionization for quantitative veratramine are m/z 410.0?295.0 and dielectric jervine?IS?m/z 426.0?114.0.Standard curve methodology validation results show that the hellebore amine linear well within the range 1-1000 ng,correlation coefficient r:0.9997,the lowest detection limit was 0.25 ng/mL.The method of precision,accuracy,recovery and stability all meet the requirements of in vivo drug analysis.The above analysis method for mouse plasma veratramine blood concentration determination and pharmacokinetic study.After the mice were fasted 12 h,respectively,a single oral administration of 1.02mg/kg and 4 mg/kg,measured at different time points after administration of plasma concentration,draw the plasma concentration-time curve.Use DAS2.0 software,using non-compart-mental pharmacokinetic model fitting device parameters.Mouse oral veratramine 1.02 mg/kg and 4 mg/kg of the main pharmacokinetic parameters?mean±SD?:T_max?h?:0.8±0.27 and 1.6±0.22 h;t_1/2?h?:1.134±0.384 and2.20±1.21 h;C_max?ug/L?:117.84±29.36 and 329.61±58.59 ug/L;AUC_0-t?ug/L*h?:255.31±43.12 and1486.78±410.27.By the maximum peak time(T_max)and half-life(t_1/2)shows that in mice veratramine rapidly absorbed and distributed rapidly.Under the curve maximum plasma concentration(C_max)and the area of medicine(AUC_0-t)with increasing oral dose increases,and prior to this study group veratramine brain damage in mice in a dose relationship coincide.