Study On Pharmacodynamic Material Of Chimonanthus Nitens Oliv. From Guizhou Province

Objective:1.The chemical constituents of the radix of Guizhou Chimonanthus nitens Oliv.have been extracted,isolated,and identified by using modern separation and extraction techniques,and the contents of toxicity and 4components of Scopolin,Fraxin,Scopoletin and Isofraxidin were compared after by different processed（drying in the shade,drying,steam,charring）.Methods:1.The radix of Chimonanthus nitens Oliv.was dried,powdered and extracted with 75%ethanol under reflux and were isolated and purified by normal phase silica gel chromatography,RP-C18 silica gel chromatography,MCI chromatography,Sephadex LH-20 gel chromatography and RP-HPLC.The structures of compounds were identified by 1D-NMR,ESI spectra.2.The Karber method was used to determine the LD₅₀ of the radix of Chimonanthus nitens Oliv.to kunming mice and evaluate its acute toxicity.3.The separation was performed on a Welch column（250 mm×4.6 mm,5μm）with gradient elution using acetonitrile（A）and water（B）as mobile phase at a flow rate of 1 m L·min^-1,and the column temperature was 30℃.The detection wavelength was 214 nm,the HPLC method was established for the simultaneous determination of Scopolin,Fraxin,Scopoletin and Isofraxidin.Results:1.Thirteen compounds were isolated from Guizhou Chimonanthus nitens Oliv..They were:（1）β-sitosterol,（2）Scopoletin,（3）Isofraxidin,（4）Scopolin,（5）Fraxin,（6）d-calycanthine,（7）Xeroboside,（8）scoparone,（9）Nitensoside A,（10）Glucosyringic acid,（11）7-O-（6-O-syringoyl-β-D-glucopyranosyl）-6-methoxy Cou-marin,（12）liri-odendrin,（13）5,6,7-Trimethoxycoumarin.Compounds 10,11,12were isolated from the genus for the first time.2.The LD₅₀of the radix of Chimonanthus nitens Oliv.to kunming mice were 4118.13 mg/kg,3733.36 mg/kg,1643.61 mg/kg by the method of drying in the shade,drying,steam three different processing methods,the 95%confidence limit was 3748.87 mg/kg-4523.76 mg/kg,3422.16 mg/kg-4072.86mg/kg,1520.90 mg/kg-1776.23 mg/kg.The charring method were>10000 mg/kg.3.The linear range of scopolin,fraxin,scopoletin,isofraxin were 0.019μg-1.856μg、0.016μg-1.616μg、0.010μg-0.920μg、0.006μg-0.624μg（r²均≥0.999 5）.RSDs of precision,stability and reproducibility tests were all lower than 2.0%（n=6）.The average recoveries were 104.47%、102.20%、101.45%、99.24%,respectively（RSD均<1.63%,n=6）.The content of the above four components were scopolin1.12%,0.90%,1.07%,0.04%,fraxin 0.87%,0.45%,0.42%,0.02%,scopoletin 0.08%,0.16%,0.08%,0.29%,isofraxidin 0.11%,0.20%,0.21%,0.25%after by different poroceed method.Conclusion:1.Thirteen compounds were isolated from Guizhou Chimonanthus nitens Oliv.,Compounds 10,11,12 were isolated from the genus for the first time,eight coumarins,1 Alkaloid,2 Lignans,1 glycoside and 1 other compound have been isolated from the plant.2.The toxicity of the Chimonanthus Nitens Oliv.were turn down after be charred.3.The HPLC method is reproducible and precise,It can be used to determine the contents of 4 active ingredients of the radix of Chimonanthus nitens Oliv.by different processed.The content of four compounds of the radix of Chimonanthus nitens Oliv.were change after processing.The specific reasons need to be further studied.The above research results enriched the medicinal material basis of Chimonanthus nitens Oliv.and provided the scientific basis for its application and development.