Study On Toxic Constituents Of Stellera Chamaejasme L.

Stellera chamaejasme L.was first recorded in “Shennong's Herbal”.Its roots can be used as medicine,spicy,toxic,into the lung,liver.Currently,many studies on the chemical constituents showed that Stellera chamaejasme L.contains a variety of flavonoids,lignins,diterpenoids,coumarins,etc.With the change of the processing method,chemical composition of the active ingredients in Stellera chamaejasme L.is also changing,resulting in the degree of toxicity also made changes accordingly.Pharmacological Studies show that water extract,alcohol extract and different kinds of processed products of Stellera chamaejasme L.have shown some anticancer activity.Some reports indicates that it has phytotoxicity,and other reports exploring the toxicity of different polarity extract parts in animals or in vitro.Therefore,we ombined some analysis methods to explore the toxic constituents of Stellera chamaejasme L.1.In this study,we developed a novel analysis method using high-performance liquid chromatography tandem time-of-flight mass spectrometer for separation and characterization of chemical compounds in Stellera chamaejasme L.We choosed Agilent Poroshell 120 EC-C18 column(2.1 × 100 mm,2.7 ?m),0.1% formic acid-acetonitrile mobile phase with gradient elution,captured mass information under ESI negative ion mode.A formula database of known compounds in Stellera chamaejasme L.was established.Finally,49 kinds of chemical composition has been identified in the total extract of Stellera chamaejasme L.,34 kinds of chemical composition has been identified in the petroleum ether extract,23 kinds of chemical composition has been identified in the ethyl acetate extract,23 chemical compositionhas been identified in the n-butanol extract.2.In this study,we screened the toxic polar part by lavaging different polar extract of Stellera chamaejasme L.in mice.In the experiment of acute toxicity,the same oral dose(10 g/kg)of different polar extract had been lavaged in mice of different groups.The results showed that the ethyl acetate group has a lowest survival rate,which indicates the ethyl acetate extract is the toxic part.After that,we lavaged mice with different concentrations of ethyl acetate extract,7 days of observation according to a detailed record of mice survial rate.According to the results,we used the PASW Statistics18.0 software to analyze the LD50(4.656 g/kg)of ethyl acetate extract acute oral toxicity in Stellera chamaejasme L.3.In this study,we used the column chromatography and high performance liquid chromatography preparation techniques to obtain the monomer compound in ethyl acetate extract of Stellera chamaejasme L.Combine their detailed mass information which was obtained from high-performance liquid chromatography tandem time-of-flight mass spectrometer and high-resolution nuclear magnetic resonance spectroscopy results,12 monomer structures were identified.They are Epiafzelechin,Scopoletin,Isolariciresinol,Lirioresinol B,Stelleranol,Daphnoretin,Chamaechromone,Neochamaejasmin A,Chamaejasmine,Isochamaejasmin,7-methoxyneochamaejasmin A and Matairesinol.4.In this study,we used serum pharmacology method to identity the toxic compounds in the mice plasma and tissues.Based on high-performance liquid chromatography tandem time of flight mass spectrometry technology,we established a method for detection of toxic components in mice plasma and tissues after oral administration.We can find the different peaks during the total ion chromatogram of blank plasma and plasma after oral administration,the total ion chromatogram of blank tissues and tissues after oral administration.In order to comfirm the different peaks,we had analyzed the monomer structures extracted in the ethyl acetate extract under the same analysis conditions.Comparing the retention time and mass information of each different peaks and monomer structures peaks,we finally found Epiafzelechin,Isochamaejasmin,Chamaejasmine,Chamaechromone,Neochamaejasmin a and Stelleranol in the plasma and tissues after oral administration.By comparing the content of six compounds in the same amount of tissues,we found this six compounds most widely distributed in the liver tissue.5.In this study,we explored the inhibition rate of six compounds to normal liver cell line L02 by using an in vitro model experiments.Epiafzelechin basically has no inhibiting effect to L02,the remaining five compounds have varing degree of inhibition in L02.6.Based on ultra performance liquid chromatography tandem triple quadrupole mass spectrometry,we established a novel analysis method for determination of Isochamaejasmin,Chamaejasmine,Chamaechromone,Neochamaejasmin A and Stelleranol in rat plasma after oral administration.We collected pharmacokinetic parameters.This method was demonstrated to be a highly special,sensitive and rapid method after a series of methodological study.This study can provoid a reference for further pharmacodynamics and toxicology studies of this five flavonoids.