Purification And Identification Of The Alkaloids In The Tobacco Material

There are many bioactive components in the tobacco, such as chlorogenic acid(CA), rutin, nicotine, solanesol etc. After being isolated and purified, these bioactive compounds can be used in agriculture, medicine, food and chemical industry. Chlorogenic acid is an important bioactive component which possesses many pharmaceutical functions, such as antibacteria, antivirus, protecting liver, cholagogue function and so on. Rutin are also used as medication for blood vessel protection or food antioxidant.Taking the waste tobacco as the raw material, the extraction and isolation of chlorogenic acid, and the purification of rutin are studied in this paper. The main achievement are as following:(1) The ultrasound-assisted extraction of principal medicinal composition(chlorogenic acid) in waste tobacco was investigated. Though testting on the major influencing factors, such as extraction solvent type, concentration, extraction time and extraction temperature, the optimum conditions were as following:using 95% ethanol as extration solvent, at 50℃, pH4, extraction for 1 hour, the ratio of solid to liquid 1:10. The extraction ratio of chlorogenic acid was 92.57%. Comparing with the efficiency of supercriticle CO2 extraction, the method of petroleum ether extraction was more suitable for removal of nicotin, the removals was 45.5%; moreover, it had the defatted function.(2) Polymaid column chromatography was used to purify chlorogenic acid. Through the dynamic experiments, the optimal preparation conditions had been identified:the concentration of sample solution, 10mg/mL; 30% ethanol eluent, eluent volume 4BV. Under this condition the purity of chlorogenic acid was 27.7%(the yield was 38.8%). After secongd purified by polyamide column(80-100mesh), the purity reached 40.7% (yield was 49.5%).Another polyphenolic product (the purity was 94.8%) was obtained by recrystallization of hot water and identified as Rutin by IR and other techniques.(3) Further experiment was studied by Ethyl acetate extraction to purify chlorogenic acid samples of polymaid resin. The chlorogenic acid concentration of the material liquid was about 12 mg/mL, phase ratio was 1/3 (V/V), extracting 2 times. The purity of chlorogenic acid sample was 62%(the extraction efficiency is 72%).(4) Finely, the purification of chlorogenic acid by silica gel column chromatography at nomal pressure was investigated. Under the condition of the rate of crude CA to silica 1:30, the purity of chlorogenic acid product reached 78.2%. On the basis of LC-MS and 1HNMR determination, four monocaffeoylquinic acids were detected. While chlorogenic acid (5-CQA,78.2%) was the major component, and other three isomers were neochlorogenic acid (3-CQA,1.9%), cryptochlorogenic acid (4-CQA,10.1%), cis-5 CQA (2.0%) respectively.Follow-up experiment by the improvement and optimization of the ratio(1:50), the product of chlorogenic acid with higher purity was obtained:the content of 5-CQA is 89.3%, while 4-CQA is 8.4%.