为快速准确地对仓储烟叶中的曲霉菌含量进行测定,通过对几种曲霉菌的 ITS区基因序列比对找出曲霉菌属遗传保守序列,以此为靶序列设计特异性引物及 Taqman探针,构建烟曲霉基因序列重组质粒为阳性标准品,并绘制标准曲线,建立了烟叶中曲霉菌含量荧光定量PCR检测方法并验证该方法具有较好的特异性、灵敏度及重复性.利用此方法对模拟霉变烟叶进行跟踪检测,结果表明:当烟叶中曲霉菌基因拷贝数在109～1011 copies·g-1 之间时,烟叶处于临近霉变的状态;大于 1011 copies·g-1 时烟叶为霉变状态;小于109 copies·g-1 时烟叶则为未霉变状态.该研究以期为仓储烟叶的霉变情况监控提供了一种更为高效灵敏的检测方法.

目的 研究栝楼不同部位(瓜蒌皮、瓜蒌子、天花粉)对卷烟烟气总粒相物(Total particulate matter,TPM)诱导的人支气管上皮细胞(BEAS-2B)增殖以及白细胞介素6(IL-6)、白细胞介素8(IL-8)、肿瘤坏死因子α(TNF-α)表达的影响.方法 采用TPM 诱导BEAS-2B 建立细胞损伤模型.实验分为空白对照组、模型组(10 μg·mL-1TPM)、红霉素组(阳性对照,10 μg·mL-1 TPM+5 μg·mL-1红霉素)及瓜蒌皮、瓜蒌子、天花粉不同浓度给药组[10μg·mL-1TPM+400、800、1 200 ng·mL-1瓜蒌皮/瓜蒌子/天花粉(正丁醇提取物)].药物作用24 h后,采用MTT法检测细胞增殖情况;采用实时荧光定量PCR法检测细胞IL-6、IL-8、TNF-α mRNA的表达;采用ELISA法检测细胞上清液中IL-6、IL-8和TNF-α的含量.结果 与空白对照组比较,模型组的BEAS-2B细胞增殖受到明显抑制(P＜0.01),细胞上清液中IL-6、IL-8和TNF-α含量明显升高(P＜0.01),细胞IL-6、IL-8、TNF-α mRNA表达明显上调(P＜0.01).与模型组比较,不同浓度组的瓜蒌皮、瓜蒌子、天花粉均可以促进BEAS-2B细胞的增殖(P＜0.05,P＜0.01);400、800、1 200 ng·mL-1瓜蒌皮组的细胞上清液中的IL-6、TNF-α含量及mRNA表达明显下调(P＜0.01),800、1 200 ng·mL-1瓜蒌皮组的细胞上清液中的IL-8含量及mRNA表达明显下调(P＜0.01);400、800、1 200 ng·mL-1瓜蒌子组的细胞上清液中的IL-8、TNF-α含量及mRNA表达明显下调(P＜0.01);400、800、1 200ng·mL-1天花粉组的细胞上清液中的IL-6、TNF-α含量及mRNA 表达明显下调(P＜0.05,P＜0.01).结论 栝楼不同部位(瓜蒌皮、瓜蒌子和天花粉)正丁醇提取物可能通过促进BEAS-2B细胞增殖,抑制TNF-α、IL-6、IL-8等炎症因子表达,从而发挥其抗炎作用.

烟草挥发性有机物(Volatile organic compounds,VOCs)含量低、组成复杂、挥发性强,同时烟草及其产品基体极其复杂,发展烟草VOCs的高效采样分析技术具有重要意义.该文综述了近年来烟草VOCs分离富集介质(如石墨烯及其复合材料、硅基材料、磁性富集材料等)及相关采样技术(如QuEChERS技术、固相微萃取技术、液相微萃取技术等)的研究进展,总结了烟草VOCs分析技术在烟草产品质量控制等方面的实际应用情况,并展望了该领域的未来研究趋势.

为探究不同滤嘴设计的卷烟爆珠单体香料成分的释放行为,分析了爆珠添加位置和滤嘴打孔情况对40种代表性单体香料成分在卷烟主流烟气中的迁移率和逐口释放量的影响.结果表明:①在迁移率方面,爆珠添加位置和滤嘴打孔情况对香味成分的释放影响显著.单体香料成分的迁移率爆珠位置在6 mm(距嘴端)处时大于18 mm处,不打孔滤嘴大于打孔滤嘴.对醇类、酮类和酯类中属于同系物的单体香料成分来说,同一成分爆珠添加位置在6 mm处情况下的迁移率与在18 mm处迁移率的比值随分子质量数的变化趋势不同;同一成分在不打孔滤嘴情况下的迁移率与打孔滤嘴迁移率的比值随分子质量数的变化趋势一致,且随分子质量数的增加而减小.②在单体香料成分逐口释放量及单位TPM逐口释放量方面,爆珠添加于6 mm处时均大于18 mm处;不打孔滤嘴单体香料成分逐口释放量大于打孔滤嘴,而打孔滤嘴单位TPM逐口释放量大于不打孔滤嘴.③沸点较低的香味成分,如己酸乙酯、庚酸乙酯、2-庚酮、2-辛酮和正戊醇等,在逐口抽吸至第4口和第5口时的释放量增幅较明显.

利用高效液相色谱-二极管阵列检测器(HPLC-DAD),建立了同时分析烟草中新绿原酸、绿原酸、隐绿原酸、东莨菪苷、咖啡酸、七叶亭、对香豆酸、阿魏酸、莨菪亭、7-羟基香豆素、芦丁、槲皮苷、山奈酚-3-0-芸香糖苷和槲皮素14种多酚化合物的方法.称取0.02 g烟末,加入5 mL 50％甲醇,超声萃取30 min,取上清液过水相滤膜后,采用Agilent SB-C18柱(3.0 mm x 150 mm,1.8|xm)进行分离,以0.05 mol/L磷酸二氢钾水溶液和甲醇为流动相,采用分段式检测,获得14种多酚类化合物在各自最佳检测波长下的信号,实现了良好分离.结果表明,14种多酚类化合物的色谱峰面积与其质量浓度呈良好的线性关系,相关系数r2>0.999 7,检出限为0.01-0.15p.g/mL,定量下限为0.03-0.50 μg/mL,日内相对标准偏差(RSD)为0.70％～3.6％,日间RSD为4.0％-6.4％,加标回收率为95.2％～109％.该方法具有良好的灵敏度、精密度和回收率,适合于烟草样品中多种多酚类化合物的同时测定.

以吸烟室为研究对象,基于CFD模拟与步进电机实验相结合,对烟气在吸烟室中的扩散进行数值模拟,并通过P IV实验测量烟气速度进行模型验证,相对误差在25％以内.最终,得到了不同送排风方式、不同烟气源量、不同新风速度下的室内烟气速度和CO质量分数分布.结果表明:侧送上排是更加有效的净化室内烟气的送排风方式;吸烟室内不同高度的烟气浓度主要取决于送排风方式、污染源的位置;增大烟气源量,会使室内气流对流加强,气流流速加快,且室内烟气浓度较高,烟客受到二手烟的危害较高;增大新风速度是降低吸烟室内烟气浓度的有效方法.

为建立烟叶霉变快速识别模型,以复烤片烟为研究对象,在高温高湿条件下进行霉变实验,获得不同霉变程度的烟叶样本.应用近红外光谱技术在4000～12000 cm-1范围内对烟叶的近红外光谱进行采集,获得烟叶样本的基础光谱数据.采用小波分解法对基础光谱数据进行解析,选择中间频率小波系数[cd4,cd5]为光谱变量,利用随机森林算法建立了不同霉变烟叶的识别模型.模型对训练集预测准确率达到93.82％,独立测试集判别准确率达到94.84％,对未霉变样品、临近霉变样品和霉变样品的判别均取得了令人满意的结果.

通过喷洒适量的甲酸溶液对烟叶进行处理,使叶片中的淀粉和蛋白质发生水解反应,从而起到有效降低淀粉和蛋白质含量的作用.50℃下,利用5％～10％的甲酸—水溶液以0.2 mL/g的喷淋量处理样品,烟叶的外观质量和微观结构无明显变化,仍具有良好的加工性能.FTIR红外光谱分析显示,甲酸处理后烟叶中的淀粉和蛋白质的特征官能团峰强度明显减弱,说明烟叶中的淀粉和蛋白质能被甲酸有效降解,且降解率与甲酸浓度呈正相关.淀粉和蛋白质的水解产物多糖、单糖和多肽、氨基酸等可转化为致香成分.感官评吸结果表明,经适宜浓度的甲酸溶液处理的烟叶杂气减少,香气增加,烟叶的品质有明显改善.

为探索烟叶中的辣感关键成分,利用凝胶渗透色谱(GPC)对一种浓香型初烤烟叶的提取物进行了感官导向分离,根据滋味特征评价结果合并辣感流份,得到了辣感特征组分;采用气相色谱质谱联用技术(GC/MS)对所定位辣感特征组分进行定性分析,鉴定出4种化合物,依据定性结果用4个标样物质进行了感官评价;对辣感特征组分中烟碱进行了定量分析,依据定量分析结果复配烟碱溶液,并分别对辣感特征组分溶液和复配溶液进行了滋味分析实验(TDA);通过空白卷烟加香考察了烟碱在卷烟中的感官作用效果.结果表明:①4种物质中只有烟碱表现出辣感特征;②辣感特征组分溶液和烟碱复配溶液具有类似的辣感强度并表现出一致的稀释倍数,说明烟碱是特征组分中辣感的主要来源,是烟叶中辣感关键成分;③烟碱与卷烟刺激性有紧密的关联,其在显著提升烟气劲头的同时,也使口腔、喉部和鼻腔的刺激明显增强.

对植物乳杆菌发酵烟叶的最佳工艺条件进行优化,研究了发酵烟叶的化学成分、香气物质和感官评价.结果表明,植物乳杆菌发酵烟叶的最佳工艺为植物乳杆菌添加量0.80‰、葡萄糖浆添加量 5.0‰、大豆蛋白肽添加量0.05‰、发酵时间 8 d,此工艺条件下,烟叶淀粉含量下降幅度最大,新增挥发性香气物质 9 种,烟碱含量下降5.58％.对比不同产地发酵烟叶后发现:外加植物乳杆菌及碳氮源可有效提高各产地烟叶的吸食品质,淀粉含量显著下降(P<0.05),水溶性总糖和还原糖含量明显提升,其中重庆产区烟叶变化幅度最大;总植物碱、总氮含量略微降低;感官评价显著提升(P<0.05),其中贵州产区烟叶提升效果最佳,总体提升 1.875 分.

The transfer rates to particulate phase of mainstream cigarette smoke and olfactory thresholds of 15 endogenous acidic aroma components in cigarette smoke were studied. The ratio of transfer rate to olfactory threshold of each component was defined as its to characterize the difference in contributions of aroma components to sensory characteristic of cigarette smoke. The results showed that:1) The transfer rate of acetic acid was the highest (36.61%), and those of the other 14 acidic aroma components ranged from 9.67% to 17.15%. 2) The flavoring contribution of isovaleric acid was the highest (169.76), and those of 3-methylvaleric acid, valeric acid, and butyric acid were in the range of 14.25-52.13, and those of acetic acid, 2-methylbutyric acid, 2-methylvaleric acid, hexanoic acid, 3-phenylpropionic acid were from 1.59 to 4.55, and those of the other components were less than 1.00. 3) The flavoring contribution integrated transfer ratio and olfactory threshold of aroma components and could comprehensively and authentically reflect the differences of flavoring components in influencing the sensory characteristics of cigarette smoke, which could be used to guide cigarette flavoring.

为研究卷烟制丝工艺中关键工序烟丝游离氨基酸的变化规律,在SIROX增温增湿和滚筒烘丝两个关键工序间进行在线取样,测定了3个批次9个样品中的氨基酸含量(质量分数),并比较了3个工序点烟丝游离氨基酸含量变化趋势.结果表明,加工前后烟丝的氨基酸含量和组成均发生了明显变化.方差分析显示,烟丝经过SIROX增温增湿工序处理,其丙氨酸、甘氨酸、缬氨酸、亮氨酸、丝氨酸和苏氨酸6种氨基酸含量显著增加(P<0.05),增加幅度3.9％～8.3％;而经过滚筒烘丝处理后,烟丝缬氨酸、亮氨酸、丝氨酸、苯丙氨酸和谷氨酸5种氨基酸含量显著降低(P<0.05),降低幅度介于1.7％～3.0％.主成分分析表明,根据9个样品游离氨基酸的分布情况,同一工序点的烟丝可聚为一类,不同工序点的烟丝能够较好地区分开.

In order to investigate sour notes in TPM of mainstream cigarette smoke with sensory-oriented analysis, the TPM was separated by gel chromatography. The characteristic sour fractions were isolated through sensory evaluation, and the sour component group with 15 compounds was identified by GC/MS method. The taste thresholds of the 15 sour components were determined, the distribution characteristics and the taste activity values (TAVs) from 12 cigarette brands were investigated, and the contribution of the 15 sour components to the overall taste of cigarettes was clarified. The sensory effect of these sour components was clarified by the sensory evaluation of cigarettes via injecting the sour components into cigarettes. The results showed that: 1) All 15 sour components contributed to the taste of cigarette. Among them, acetic acid and lactic acid contributed the most. The TAVs of lactic acid from the different brands of cigarettes varied greatly. 2) Some sour components also presented sweet taste and astringent feeling. 3) The application of sour components reduced the irritation of cigarette smoke, improved its aftertaste, and promoted the sour taste, sweet aroma note and sweet taste of cigarette smoke.

The aroma components of tobacco leaves are important indexes to evaluate the quality of tobacco leaves.Through the study of the variety and content of aromatic constituents in the aging process of tobacco leaf,the quality of tobacco leaf was monitored and the best time to control tobacco leaf aging was monitored,and the economic benefits were improved.Tobacco leaves have many aroma compositions,complex compositions and low content.It is very important to adopt appropriate pretreatment methods and analytical methods.This paper briefly introduced the changes of aroma components in tobacco leaves and their aging process,with emphasis on previous treatment,analysis and analysis of chemical metrology,also reviewed the research progress in the analysis of aromatic components in tobacco leaves,and compared the principles,advantages and disadvantages of pretreatment methods (distillation extraction,solid-phase microextraction and liquid-liquid microextraction) and analysis methods (gas chromatography,gas chromatography-mass spectrometry,full two-dimensional gas chromatography-flight time mass spectrometry).Finally,it summarized the development trend of aromatic composition analysis in tobacco leaves.

Objective To study the anti-alcoholism effect of Puerariae lobatae radix, Puerariae lobatae flowers, Hovenia dulcis thunb mixture on acute alcohol poisoning in mice.Methods A model of acute alcoholism in mice was established by intragastrical administration of alcohol.The best ratio of each component in the mixture was identified by observing the sobering effect of the mixtures on mice.The best ratio mixture was divided into low dose group and high dose group.The mice tolerance time, drunken time, the ethanol concentration in the blood and the alcohol dehydrogenase activity in liver were measured to study the anti-alcoholism effect of the mixture.Results The Puerariae lobatae radix, Puerariae lobatae flowers, Hovenia dulcis thunb mixture not only increased the tolerance time and decreased the drunken time of mice, but also significantly reduced the ethanol concentration in blood and improved the activity of alcohol dehydrogenase in liver.Conclusion This study found that Puerariae lobatae radix, Puerariae lobatae flowers, Hovenia dulcis thunb mixture has anti-alcoholism effect on acute alcoholism in mice.

The enzymolyzed products of Moringa Oleifera seed were prepared by aqueous enzymatic extraction.The protein recovery and l,l-diphenyl-2-picry lhydrazyl (DPPH) radical scavenging activity of the enzymolyzed products were selected as indexes to optimize the enzymatic hydrolysis conditions.Single factor experiment showed that the enzymolyzed products obtained by Alcalas 2.4L and Ns 37071 had the highest protein recovery and DPPH radical scavenging activity.The protein recovery were 45.65％ and 47.65％,respectively,and the DPPH free radical scavenging activity were 99.88 and 93.89 μmol Trolox equiv/100 g,respectively.Further,the optimal enzymolysis conditions confirmed by response surface methodology were as follows:the total amount of enzyme 2.60％,Ns37071 accounted for 46％ of the total amount of enzyme,and enzyme time 24 hours.The protein recovery was 63.69±2.94％,and DPPH radical scavenging activity was 139.38±0.96μmol Tmlox equiv/100 g.The study in this paper showed that the aqueous enzymatic extraction could efficiently hydrolyze Moringa oleifera seed,and enzymolyzed products had better protein recovery and DPPH radical scavenging activity.

In order to investigate the alleviation effect and mechanism of Puerariae flos on acute alcoholism, a test cigarette was prepared by adding a certain amount of Puerariae flos into cut tobacco. According to the fumigation therapy theory and action mode of traditional Chinese medicines, experimental mice were treated by treatment 1 (gastric lavage with alcohol + mainstream cigarette smoke exposure) and treatment 2 (mainstream cigarette smoke exposure + gastric lavage with alcohol) separately, then the antialcoholism effects and the variations of alcohol concentration in blood and alcohol dehydrogenase activity in liver of mice were investigated. The results showed that: 1) Comparing with control groups, the drunkenness time of treatment 1 group significantly reduced when a high dosage of Puerariae flos was added into cigarette; for treatment 2 group, alcohol tolerance time remarkably increased, while drunkenness time decreased with the increase of Puerariae flos adding rate. 2) Increasing Puerariae flos adding rate raised the alcohol dehydrogenase activity in liver, while decreased the alcohol concentration in blood obviously along with the proceeding of time. Mainstream smoke of cigarette pretreated by adding Puerariae flos into cut tobacco has an obvious alleviation effect on acute alcoholism in mice.

To discuss the extraction technology and classified polysaccharide molecular weight distribution,pomelo peel polysaccharide was extracted by enzyme extraction and organic solvent.Distribution of molecules weight was carried out by GPC chromatogram.The optimization of polysaccharides from pomelo peel by the enzyme extraction was:enzyme dosage 1.5％,reaction time 120 min,solid to liquid ratio of 1 ∶ 25,enzymolysis temperature 50 ℃.The total polysaccharide content of pomelo peel was 10.39 g/100g powder.The crude polysaccharide was then separated into 3 grades based on its content:grade A was 5.6 g/100g powder,grade B was 1.9 g/100g powder and grade C was 1.8 g/100g powder.The purity of coarse polysaccharide of different grades was different.The molecular weight distribution tests showed that the lower the ethanol concentration,the higher the molecular weight.Polysaccharide(a) molecular weight in different grades were:24 100 Da in grade A,11 700 Da for grade B and 2 130 Da for grade C.

本文以柚子果皮为材料,采用热水提取结合低温醇沉制备柚皮多糖.通过扫面电镜和化学方法分析研究六种烟叶载体物理结构和化学成分的差异,并研究柚皮多糖在不同烟叶载体上保润动力学.扫面电镜表明不同载体表面光滑度不同,表皮气孔数量也不相同,上部叶B3F表皮气孔排布要比中部叶C2F致密一些,中部叶C2F表皮气孔排布又比下部叶B1F要致密一些.化学成分有机酸含量DY2含量最高为28.1％,DY3最低为16.2％.多酚含量由2.2 mg/kg增加至5.58 mg/kg,品种间有显著差异.总糖含量最高是C2F为26.37％,显著高于其它几种,烟碱含量DY1最低为2.82％,B3F最高为5.12％.保润特性表明:自身保润特性呈现一定的差异,柚皮多糖在六种烟叶载体上保润动力学呈现不同的变化趋势,对B3F载体的保润效果最佳,对B1F载体的保润效果相对较差.

本文以柚子皮为原料,通过响应面优化了柚子皮多糖热水浸提工艺,并对柚子皮多糖保润特性进行了研究.结果表明:响应面优化柚子皮多糖热水浸提工艺为:浸提温度80℃、液料比30:1(mL/g)、浸提时间2.5h和浸提次数2次,多糖提取率达到6.45％.通过6种烟草的柚子皮多糖与丙二醇在失水率到达平衡时的对比,柚子皮多糖对不同烟草的保润呈现不同,其中柚子皮多糖保润剂对B3F的保润效果最佳,对B1F的保润效果相对较差.