Preparation The Chemical Reference Substance Of Nitidine Chloride

Objective:To obtain the optimal process for separation and purification the Nitidine Chloride by cationic resins, and to obtain the Chemical Reference Substance by refining the crude product with column chromatography and crystallizationMethod:The content of total alkaloids from Zanthoxylum nitidum was determined by UV spectrophotometry, and the content of the Nitidine Chloride was determined by HPLC. Identified the structure of the product by IR,1H-NMR,13C-NMR and MS spectra analysis.Static adsorption-desorption methods were adopted to select the best cation exchange resin, and the separating efficiency was evaluated by adsorption and desorption rates of cation exchange resins. The process such as adsorption , desorption and regeneration were optimized with the single-factor experiments and orthogonal experiments. The crude product was refined with ultrasonic wave in solvent, column chromatography and crystallization in turn to obtain the Chemical Reference Substance. Result:The optimum separating process with the cation exchange resin:The Ls006 cation exchange resin had the best separating efficiency for Nitidine Chloride in nine types of cationic resins.Extracted the Zanthoxylum nitidum(Roxb.)DC. granule with 60% alcohol for 3 times. The extraction efficiency of total alkaloids and the Nitidine chloride were 86.83 and 66.54%, respectively. The extract mixture was used as loading solutions.The Ls006 cation exchange resin and loading solutions were stirred at room temperature, desorbed with stripping liquid for 4 times, merged all the filtrate and gained the crystal by centrifugalize in 2000 rpm. At last,the average adsorption and desorption rates were 83.70% and 89.41%,respectively.The yield of Nitidine chloride was 51.22%, the product purity was over 80%.The cation exchange resin must be regenerated after being used 5 times.The process of purification and refining:Treated the crude product by mean of ultrasound wave, silica gel column chromatography and crystallization in sequence. At last, the yield of Nitidine Chloride was up to 39.02% and the purity was over 98%.At last, the yield of Nitidine Chloride of the whole process was 19.99%. The melting point, IR, 1H-NMR, 13C-NMR and MS Spectroscopy information of the product were in accord with the target compound. The product purity was over 98%.Conclusion:The melting point, IR, 1H-NMR, 13C-NMR and MS Spectroscopy information of the product are in accord with the target compound. The purity of final product is over 98% which has met requirements of the reference substance for identifying.The Ls006 cationic resin has the best efficiency for extracting and seperating Nitidine Chloride.It is obviously that its adsorption and desorption rates,and the purity of the product of Nitidine Chloride are high.This process is low production cost, high efficiency and easy to operate. What is more, this process could avoid using large amount of Chloroform for extracting, it is economical and environmental protection, so it is worth in industrial application. The Chemical Reference Substance is preparated after treating the crude product by mean of ultrasound wave, silica gel column chromatography and crystallization in sequence.It is so easy to operate and get a so remarkable effect that it can be apply to multitudinous preparation.