Study On The Extract Of Radix Aconiti Kusnezoffii Standard

Radix Aconiti Kusnezoffii (Cao-Wu) is the dried tuber of Aconitum kusnezoffii Reichb. It has the characteristic and action of relieving rheumatic conditions, warming the channels, alleviating pain and high toxicity. It can be used for rheumatic and rheumatoid arthralgia, precordial and abdominal pain with cold sensation and anesthesia. It is used widely but sometimes causes poisoning events. Because the quality of the crude drug from various resources was usually different, due to the difference of species, district, process method, quality evaluation, etc. So it's necessary to develop a standard extract of Radix Aconiti Kusnezoffii (SEAK) and its preparations. In the dissertation, the aspects of quality evaluation, extraction technology, hydrolytic kinetics of aconitine, preparation technology,acute toxicity and analgesic effect for SEAK were studied. Also the quality of Radix Aconiti Kusnezoffii and Radix Aconiti (the dried tuber of A. carmichaeli Debx.) from various resources was compared.The methods for the determination of Aconitum alkaloids in Radix Aconiti Kusnezoffii were studied. Volumetric analysis with the indicator of methyl red-bromcresol green and colorimetric analysis with acid dye of bromecresol green were used for the assay of total alkaloids. Colorimetric analysis of hydroxamic acid-Fe was used for the assay of total ester-diterpene type Aconitum alkaloids. HPLC method, sensitive and accurate, for the determination of three kinds of diester-diterpene type Aconitum alkaloids (aconitine, hypaconitine and mesaconitine) and HPLC method for the fingerprint were established. Radix Aconiti Kusnezoffii and Radix Aconiti from various recourses were also determined by the above methods. The results showed that the content of total alkaloids and the similarity of the HPLC fingerprint were affected significantly by the factors of species, district, process method, etc. The conditions of extraction process including primary extraction, flocculation condition of chitosan, and purification technology of macroporous resin for SEAK were optimized by orthogonal design. According to the results of 11 batches of SEAK, the content (average content 94.18%) of total alkaloids, the content and the ratio of three kinds of diester-diterpene type, and the HPLC fingerprint could besteady by the established extraction process. The standard extracts of Radix Aconiti Kusnezoffii (SEAK) and Radix Aconiti from various resources were also prepared and evaluated in the same manner. The results suggested that the resources of the crude drug and the extraction process with good repeatability were the important factors for the stabilization of SEAK.The mechanism and kinetics of the hydrolysis of aconitine were studied in this paper. The results showed that in addition to benzoylaconitine, a compound with the molecular of 585, which might be pyraconitine or 16-epi-pyraconitinin, was found in the hydrolyzates of the first-step hydrolytic degradation of aconitine. The results of the hydrolytic kinetics of aconitine investigated by HPLC-UV showed that the reaction was catalyzed by OH- ion in the pH range from 2.0 to 7.0, the rate constants of hydrolysis depended significantly on the value of pH and temperature, and the rate constant increased with the increment of pH value while it was stable in the pH range from 2.0 to 4.0 and increased greatly as pH>5.0. Furthermore, the rate constants of the first-step hydrolytic degradation of aconitine (k1) and t0.9 were calculated by Arrhenius's equation, which provided the important information for the stability of aconitine. Based on the modern transdermal theory, the transdermal permeability of SEAK was studied for the first time. The results showed that the flux of SEAK through the back skin of SD rat was neglectable without penetration enhancer. Ethanol, Azone, oleic acid, isopropyl myristate, and eucalyptus oil could increase the permeation of SEAK, and the mixture of ethanol and eucalyptus oil was the optimum enhancer combination. On the ba...