【作者】 吴维佳；

【导师】 齐绍武； 朱茂祥；

【作者基本信息】 湖南农业大学 ， 烟草科学与工程技术， 2010， 硕士

【摘要】 卷烟是一种特殊食品,而烟用香精香料是这一特殊食品的添加剂,对于卷烟风格特征的形成具有不可或缺的作用,其配方更是卷烟工艺中的核心技术,烟用香精香料的安全性已备受消费者和烟草企业的关注。香精香料加入到卷烟后的毒性很值得研究和探讨。细胞毒性试验和遗传毒性试验作为有害物质危害性评价的重要手段被广泛应用。本研究采用CORESTA(国际烟草科学研究合作中心)推荐的三项体外毒理学评价方法MTT(四甲基偶氮唑盐比色法)细胞毒性试验,微核试验和AMES试验(鼠伤寒沙门氏菌回复突变实验)对卷烟中的香精香料及其拆分组分进行了评价。本论文的主要研究内容与结果如下：1、香精香料及其拆分组分未添加至卷烟之前,随物质浓度的增大细胞的存活率均出现下降,表现出剂量效应,呈负相关。全香精细胞毒性明显高于全香料(p<0.01),全香料的10个拆分组分在最高所试剂量(50ml／L)未表现出细胞毒性,全香精的10个拆分组分细胞毒性存在差异,其中3＃、5＃、7＃和8＃(此编号为拆分组分的编号,具体成分为商业秘密,在此不给出)香精组分的细胞毒性较低,其他几种香精组分的细胞毒性相当,但所有香精拆分组分的细胞毒性均明显低于全香精(p<0.01)。2、香精香料及其拆分组分添加至卷烟后,随CSC(卷烟烟气凝集物)浓度的增大细胞的存活率均出现下降,也表现出剂量效应,呈负相关。与对照烟比较,添加香精的卷烟,细胞毒性无明显增加,与全香精比较,10个香精拆分组分的细胞毒性无差别。与对照烟比较,添加香料的卷烟,细胞毒性也无明显增加,与全香料比较,10个香料拆分组分的细胞毒性无差别。3、对香精香料及其拆分组分的卷烟烟气CSC进行微核分析,流式细胞仪检测结果显示经卷烟烟气染毒细胞微核率高于未染毒正常细胞,全香精5#、10＃拆分组分比其他组分微核率稍高,但是全香精的微核率与未添加香精空白对照烟无差异。全香料3＃、4＃、5＃拆分组分比其他组分微核率稍高,但是全香料的微核率明显低于未添加香料空白对照烟。提示某些拆分组分增加了卷烟的遗传毒性,提示在烟用香精香料配伍时可少用或者不用毒性较高的组分,保证其在卷烟应用时的安全性。4、对香精香料及其拆分组分的卷烟烟气CSC进行致突变分析,CSC的加入量与突变菌落数呈极显著正相关,存在线性剂量关系,即随着CSC剂量的增加,其致突变性也相应增强。与空白对照烟比较,全香精、全香料及其拆分组分致突变性无显著差异。整个实验结果提示该香精香料在卷烟中的应用是安全的。更多还原

【Abstract】 Tabacco essence and flavor, additives for the special foodstuff—Cigarette, are essential to form the cigarette style, and the prescription of them is the key technology in cigarette technics. The safety of the tabacco essence and flavor has raised much attention of the consumers and tobacco companies. The toxicity of the essence and flavor in tobacco is worth to research and discuss. The tests of Cytotoxicity and Genetic toxicity are widely used as important means to evaluate the dangers of the detrimental agents. This study evaluated the tabacco essence and flavor and it’s splited components by Three in vitro toxicology methods(recommended by CORESTA)、MTT Cytotoxicity Test、Micronucleus Test and AMES Test.The results were as follows:1、Before injection of essence and flavor and their splited components into the cigarettes, the cell survival rate declined as the detriments increased, which showed dose-dependent trend of the cell. The cytotoxicity of whole essence was apparently higher than the whole flavor(p<0.01). The ten splited components of whole flavor showed no cytotoxicity in the maximal concentration (50ml/L) of the test dose. The cytotoxicity of ten splited components of whole essence was different. The 3#、5#、7#、8# components had lower cytotoxicity and the rests had relatively same level of cytotoxicity. However, the cytotoxicity of all splited components of essence were obviously lower than the whole essence (p<0.01).2、After injection of essence and flavor and their splited components into the cigarettes, the Cell survival rate also decreased when the dosage of CSC(cigarette somking condensates) increased, and showed a dose-dependent trend as well. Compared with blank cigarettes, the essence or flavor injected cigarettes had no obvious cytotoxicity increase and the cytotoxicity of the ten splited components of whole essence or flavor had no difference in comparation with whole essence or flavor separately.3、The cigarette somking condensates(CSC) of essence and flavor and their splited components injected cigarettes were evaluated by Micronucleus Test. The Flow Cytometry showed that:the cell micronucleus rate of cells poisoned by CSC was higher than the unpoisoned ones. The Cell micronucleus rate of splited components of whole essence 5#、10# were a little higher than other components, but the cell micronucleus rate of whole essence had no difference with the blank cigarettes. The Cell micronucleus rate of splited components of whole flavor 3#、4#、5# were a little higher than other components, but the cell micronucleus rate of whole flavor was obvious lower than the blank cigarettes. The results shows that some splited components increased the descendiblity toxicity of cigarettes, therefore, we could use less or even no the higher toxicity components to ensure the safety of cigarettes.4、The cigarette somking condensates(CSC) of essence and flavor and their splited components injected cigarettes were evaluated by the mutagenicity AMES test. The increase of the CSC resulted in a higher mutagenicity of the Revertant Clonies, which showed an apparent positive correlation and a linear does effect between them. The cigarette somking condensates of the whole essence and flavor and their splited components injected cigarettes had the same level of mutagenicity as blank cigarettes. All the experiment results demonstrated that it’s safe to use essence and flavor in cigarettes.更多还原