Analysis Of Active Components And Research On The Toxic Testing Method Of Kusnezoff Monkshood Root Of Yunnan

Kusnezoff monkshood root of Yunnan (Aconitum Transsectum Diels) , alternate name Da cao wu, Xiao hei niu, is widely distributed northwest Yunnan of China. This study used the wild type and cultivated type kusnezoff monkshood root as test material, cooked this material for gradual detoxification, analyzed the content and structure of the Kusnezoff monkshood root active components by HPLC-UV and HPLC-MS. In order to evaluate the toxicity and efficacy, and according to the evaluation result to research the toxic detection method.1 Analysis about the main composition of kusnezoff monkshood rootThis study used volumetric method and colorimetric method to analyze the total alkaloid, free amino acids, soluble sugar and soluble protein in kusnezoff monkshood root with different varieties, different treatment methods and different steaming time. The result is that soluble sugar comprises 25 percent to 52 percent, soluble protein comprises 0.45 percent to 0.89 percent, free amino acids comprises 0.45 percent to 0.96 percent of net weight of kusnezoff monkshood root, the content of total alkaloid in kusnezoff monkshood root is 0.023mol/g in wild type and 0.020mol/g in cultivated type. According to the result, soluble sugar is the main composition in this material; the Content of total alkaloid in wild type Kusnezoff monkshood root is higher than the cultivated type; the steaming time and treatment methods have little effect on the content of total alkaloid.2 Analysis about the toxic ingredients of kusnezoff monkshood rootBy using the detector of HPLC-UV and HPLC-MS to analyze the toxic ingredients which are extracted by organic solvent extraction method in kusnezoff monkshood root, using this result for quantitative and qualitative analysis. The result shows us, the retention time of alkaloids in non-cooking kusnezoff monkshood root are 10.9min, 17.8min, 26.1min, 37.0min, 56.8min, 71.9min, and their Molecular Weight are 421, 463, 433, 601, 541, 659; the retention time of alkaloids in cooking kusnezoff monkshood root are 10.9min, 15.4min, 17.8min, 23.1min, 26.1min, 56.8min, 70.7min, and their Molecular Weight are 421, 391, 463, 617, 433, 541, 659. According to the result, the retention time of 10.9min, 23.1min, 70.7min of alkaloids are talatisamine, 8 deacetylyunaconitine, yunaconitine, and the yunaconitine is the diester-diterpenoid alkaloid that shows highly toxicity, the yunaconitine content is very high in kusnezoff monkshood root. This study finds that there are not differences in the alkaloid ingredients between the non-cooking wild type and the non-cooking cultivated type kusnezoff monkshood root; also, there are not differences in the alkaloid ingredients between the cooking wild type and the cooking cultivated type kusnezoff monkshood root, but there are significant differences in the content of four kinds of alkaloid between this two kinds cooking material. The content of high molecular weight alkaloid is high in the non-cooking kusnezoff monkshood root; but the samples which are cooking for over 6h on atmospheric pressure are basically contains no high molecular weight alkaloid, so we can holds that these samples are detoxification. The alkaloid ingredients and the content of them change hardly in earlier stage, but the content of them change gently in later stage, this result express that cooking over six hour on atmospheric pressure or over four hour on elevated pressure can get the detoxification efficiency. It was concluded from above results that the cooking time is the most important detoxification methods. In short time cooking under the elevated pressure can get the same detoxification efficiency in long time cooking under normal pressure.3 Research the toxic detection method for kusnezoff monkshood root(1) The reagent selection testIn the W extracts of the non-cooking kusnezoff monkshood root, we put the A, B, C, D and E in, and observe the result of color reaction. The result shows that the reagent of concentrated D and high concentration W are the best detection reagent.(2) The optimization reagentThe result of full wavelength scanner of present-color material shows that the maximum absorption spectra is 522nm; The result of colorimetric determination show us that the color material has good stability. Using orthogonal test to determine the best detection conditions for laboratory detection, the best conditions are that the W concentration is 90%, the D concentration is 0.08mL/mL, the addition amount of kusnezoff monkshood root is 0.8g, the temperature of the reaction is 20℃.(3) The toxic detectionAccording to result of color reaction in kusnezoff monkshood root, the color is red in condition of non-cooking root, and the red color is becoming light with the cooking time becoming long. With the colorimetric determination, the results are that the absorption value of non-cooking root is 0.706, the absorption value of cooking root is 0.115, these results of the color reaction corresponds to the results of HPLC detection. Through the above analysis, the toxic detection method can handily, fleetly and effectively detect the detoxification of kusnezoff monkshood root.