Study On Decreasing Toxicity And Increasing Efficacy Of Acointe Based On Biotransformation

Objective To establishing a HPLC method for simultaneous determination ofaconite, hypaconitine and mesaconitine in radix aconiti carmichaeli, studing on theHPLC fingerprint of radix aconiti carmichaeli and processing radix aconiticarmichaeli, select the microorganisms which can biotransformation of radix aconiticarmichaeli, structure identification of the new chemical constituents frombiotransformation by PHLC-MS, study on the reducing toxicity and enhancing effectsof biotransformation radix aconiti carmichaeli by pharmacology and pharmiceffectiveness tests.Method①To take the aconite, hypaconitine and mesaconitine as references, theHPLC was applied to determine the content of aconite, hypaconitine and mesaconitine.②The similarity evaluation system for chromatographic fingerprint of TCM(2004Aedition) was used to evaluate the similarity of the10batches of radix aconiticarmichaeli,and10batches of processing radix aconiti carmichaeli. Compare theHPLC fingerprint of10batches of radix aconiti carmichaeli,10batches of processingradix aconiti carmichaeli, adix aconiti carmichaeli betweent and processing radixaconiti carmichaeli.③Take biotranformation with radix aconiti carmichaeli by solidfermentation, select the microorganisms which can biotransformation of radix aconiticarmichaeli to microorganism growth, alkaloids content and new chemicalconstituentsas the evaluation index.④Structure identification of the new chemical constituentsfrom biotransformation by PHLC-MS, study on the reducing toxicity and enhancingeffects of biotransformation radix aconiti carmichaeli by pharmacology and pharmiceffectiveness tests, provide a theoretical basis and practical feasibility forbiotransformation with radix aconiti carmichaeli.Result①Aconite in radix aconiti carmichaeli was linear in the ranged of0.17～2.72μg, y=195555x－11910, R2=0.9995, and Hypaconitine was linear in the rangedof0.13～2.08μg, y=335727x－16744, R2=0.9996, and Mesacontine was linear in theranged of0.15～2.40μg, y=375160x－20194, R2=0.9996, The average recoveries ofaconite, hypaconitine and mesaconitine were100.04%,99.99%,100.09%, themethodological study showed that the method is simple, accurate and stable reliability.②The similarity evaluation system for chromatographic fingerprint of TCM (2004Aedition) was used to evaluate the similarity of the10batches of radix aconiticarmichaeli, and10batches of processing radix aconiti carmichaeli, the results showthat the radix aconiti carmichaeli fingerprints were feasilble, which could be used forindex detection and quality control.③Select the microorganisms which canbiotransformation of radix aconiti carmichaeli to microorganism growth, alkaloidscontent and new chemicalconstituents as the evaluation index, the results show thatthere were grew well, contents of alkaloids increased, formed new chemicalconstituents with mold, and other strains were not to be.So Aspergillus niger is confirmed the biotransformation strain of radix aconite carmichaeli, and one of thenew chemical constituents was identified preliminarey to be benzaconine with lesstoxicity and high drug effect.④Test of reducing toxicity and enhancing effects isshow that the radix aconiti carmichaeli of biotransformation were reduced toxicity,increased effects, it is can be less toxicity and high drug effect, And the result offerssome basis for futther identified of benzaconine.Conclusion The HPLC method was reliable for the determination of aconite,hypaconitine and mesaconitine in radix aconiti carmichaeli; the fingerprint methodwas available, and can be used to evaluate the quality of the radix aconiti carmichaeli.The radix aconiti carmichaeli of biotransformation with Aspergillus Niger could bereduced toxicity, increased effects, provide a theoretical basis and practical feasibilityfor next research of processing method with radix aconiti carmichaeli.