Study On Determination Of Phenolics In Tobacco And Cigarette Smoke And Its Application

A new method for determination of polyphenols in tobacco leaf with solid phase extraction-high performance liquid chromatograpgy-mass spectrum (SPE-HPLC-MS) were studied. Polyphenols can be extracted from tobacco leaf with 80% methanol by heating circumfluence, then subjected to degrease by Waters Sep-Park-C18 solid phase extraction cartridge. Main polyphenols can be separated on a Waters Nova-Pak-C18 chromatogram column (3.9×150mm),by methanol and 1% acetic acid solution as mobile phase at flow rate of 0.5mL min-1 with injection volume of 10 u L. Qualitative analysis electrospray ionization mass spectrometry (ESI/MS) method was used. Standard sample and samples were determinated at the maxmum absorption wavelength of 345nm,and their peak areas were calculated. Compared with traditional method, this method shortened the time of preparation sample greatly. The relative standard deviation and recovery were 1.31%-2.36% and 94%-103% respectively. The regulations of main polyphenols in tobacco between the variety tobacco, the stalk position of tobacco leaf, the amount of nitrogen-containing fertilizer and the modulation progress of tobacco were investigated by use of advanced SPE-HPLC-MS. The results were as follow:the contents of total polyphenols in Hongda breed are higher than other breeds; for the different stalk position of the same breed, the content of total polyphenols in top leaves in higher than in middle and bottom; proper amount of appliled nitrogen-containing fertilizer (such as 6-8kg/mu) is benefited to the accumulation of polyphenols; in the different types of tobacco, the content of polyphenols in flue-cured tobacco is highest, but burley tobacco is lowest; the content of total polyphenols is not obviously changed during the course of fore-and-aft tobacco accumulation.According to the domestic and foreign review, two new methods were appilied in the determination of phenolic compounds in smoke. The total phenolic contents were determined through auto-vapor distillation and spectrophotometer at 460nm wavelength. Recovery and ralative standard deviation of this method were 96%-105% and 1.1%-1.4% separately. Contrary on the way of common water vapor, it is anexacter result, higher precision, simple operation and shorter time. In addition, seven phenolic compounds (catechol, hydroquinone, resorcinol, phenol, o-cresol, m-cresol and p-cresol) tested by the way of high performance liquid chromatograph at the same time. Experimental condition is as follow : Waters Symmetry?C18 reversed phase bonded silica gel chromatogram column (3.9 X 150mm), methanol and 0.05molL-1 potassium dihydrogen phosphate solution gradient eluting as mobile phase at flow rate of 0.5mL min-1 with injection volume of 10 u L. Contrary on the reported method, its operation of simple treatment is simpler, analytical time is shorter, the accordance is better and the price of flow phase is lower.Test thirty-five phenolic compounds in forty-seven brands cigarette smoke by GS-MS. The correlations between phenolic compounds and sensory properties were studied by SPSS(Statistics Package for Social Science) in the computer. The mathematics models were set up by means of correlation and multi-variable linear regression analysis. The results showed that most phenolic compounds are indirect to sensor quality and only a few ones are direct. So some measurements must be taken to reduce phenolic compounds which are harmful to human body.Acording to the corelation analytical results, polyethonolisomers as an additives of reducing phenolics was synthesized by reaction between polyethonol, catalyzer made by us and compound A. reaction conditions are 120 degree in reaction temperature, 0.1 to 0.3 precent in the catalyzer does and the rate of one to one. This serial additives were applied in four brand cigarettes from three companies of Yunnan privence. The two above mentioned methods of determinating phenolics in smoke were appilied in four types of cigarettes and the reduced percentage of the contents of phenolic compounds was calculated. Sensor...