【作者】 孟冬玲；

【导师】 杨亚玲； 缪明明；

【作者基本信息】 昆明理工大学 ， 应用化学， 2004， 硕士

【摘要】 建立了固相萃取-高效液相色谱-质谱法测定烟草样品中多酚类物质的新方法。用80％的甲醇溶液回流提取烟叶中的多酚物质，然后用Waters Sep-Park-C₁₈固相萃取柱分离脱脂；以Waters Nova-Pak-C₁₈色谱柱（3.9×150mm，5μm）为固定相，甲醇-1％醋酸溶液为流动相；流速为0.5mLmin^-1；线性递增（减）的梯度洗脱，进样体积10μL，并用电喷雾质谱对其进行辅助定性；标样和烟草样品在345nm波长处提取色谱图计算峰面积。此方法的优点主要是采用固相萃取柱对样品进行前处理，使干扰组分保留在小柱上而酚类物质不保留，而且对环境不造成污染。它比传统的方法脱脂时间缩短约2小时，相对标准偏差为1.31％-2.36％，回收率为94％-103％。在该方法的应用中，研究了烟叶部位、烤烟品种、施肥量、烟叶种类以及调制对多酚类物质的影响。结果表明：同一烤烟，上部烟叶多酚含量比下部高；不同烤烟品种中，红大品种绿原酸含量高；中等程度施肥量对多酚积累最有利；烤烟中多酚含量最高、白肋烟中最低；调制对多酚类物质的影响不大。 对于卷烟烟气中酚类物质的测定，综合了国内外的研究进展，主要采取了两种方法。自动水汽蒸馏分光光度法测定卷烟烟气中的总酚，检测波长为460nm，回收率为96％-105％，相对标准偏差为1.1％-1.4％。与普通水蒸汽分光光度法相比，此方法蒸馏时间缩短约55分钟，整个系统密闭性好，气流稳定，从而使测定结果更准确、精度更高。其次，用高效液相色谱荧光检测法测定了卷烟烟气中的七种酚类物质即邻苯二酚、间苯二酚、对苯二酚、苯酚、邻甲酚、间甲酚和对甲酚。实验条件为：色谱柱为Waters公司的Symmetry（?） C₁₈反相键合硅胶柱，流动相为甲醇-磷酸二氢钾缓冲液，流速为0.5mLmin^-1，分析时间为30min，回收率为96％-101％，相对标准偏差为1.2％-2.6％。此方法的优点是无需衍生化，样品处理时间缩短3.5小时，分析时间缩短一半，便于批量处理，而采用甲醇作为流动相比乙腈在成本上降低了一半。 另外，本文对酚类物质与感官评吸的相关性进行了研究。首先采用GC-MS联用仪测定47种卷烟烟气中的225种成分，筛选出其中的35种酚类物质作为分析数据，然后采用SPSS统计分析软件进行多元线性回归分析并建立相关性数学模型。感官评吸指标与酚类物质基本呈负相关，只有少量酚类物质呈正相关。均负相关的是2，5-二甲基苯酚和3-乙基苯酚；除刺激性外，其他5项均与4-吡啶酚、4-乙烯基苯酚负相关；除刺激性和和香气外，均与2，4-二甲基苯酚负相关；谐调、刺激性和综合得分均与苯酚、4一乙基苯酚负相关:香气、刺激性和余味均与4一乙烯基一2-甲氧基一苯酚负相关:香气、刺激性与对苯二酚负相关:刺激性和余味与1，2一苯二酚负相关。均正相关的是4一甲基苯酚、2，3一二甲基苯酚;除谐调外，均与2一甲基苯酚正相关:除刺激性和谐调外.均与2.3，5一三甲基一l，4一苯二酚正相关、余味和谐调均与2，6一二甲基苯酚正相关;香气和余味均与香兰素正相关;而2，4一二甲基苯酚只与刺激性正相关.这些相关性对感官评吸人员和实际生产应用有指导作用. 根据酚类物质的危害及其与感官评吸的相关性结果，本文首先合成了降低卷烟烟气中酚类物质的CW系列添加剂，即由聚乙二醇、自制催化剂和化合物A合成的聚乙二醇衍生物（Cw系列嘴棒添加剂）。反应条件为:反应温度120℃，催化剂用量为0.1%一0.3%，反应配比1:1。把筛选出的CW4和CW6添加剂应用到省内3家卷烟厂四种牌号的不同等级卷烟上，并采用己建立的两种方法测定出上述四种卷烟烟气中酚类物质的含量，并计算其下降的百分比。CW添加剂能使苯酚下降10%一45%，邻苯二酚则不超过20%。除样烟B的CW4外，其它样烟的间苯二酚均呈下降趋势，为66%一99%，对苯二酚在样烟D中均下降，以CW4效果最好，为10.92%。毒性较大的邻、间、对甲酚在样烟A中呈上升趋势，而在其它三种品牌中均有不同程度的下降，为30%一56%。总酚下降为6一26%。为了保证卷烟的总体指标合格，对加入添加剂后制成的卷烟进行了一系列检测.感官评吸是通过人体的感觉来评价卷烟的香吃味，判断刺激性的强弱，进而知道烟气中酚类物质含量的多少。采用前面建立的方法测定烟气中总酚和七种酚，并分别计算酚类物质下降的百分比。以上五种检测结果均表明:合成的CW系列添加剂能降低上述卷烟烟气中的酚类物质，CW6使中低档样烟A、B、C中的酚类物质下降幅度最大，效果最显著。而对优质的样烟D而言，CW4系列添加剂比CW6系列的效果要好。此添加剂优点是环保性添加剂，易合成且工艺条件易控制。为此类添加剂的工业化生产和在烟草中的应用提供了有力的理论依据。更多还原

【Abstract】 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更多还原