利用生物活性材料降低卷烟主流烟气关键有害成分含量的研究
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摘要
烟草消费所导致的健康问题日益突出,低焦油、低危害卷烟已经成为国际烟草行业主要发展方向。本论文系统研究了烟用普通醋酸纤维滤嘴及二醋酸纤维丝束的生产工艺,以生物活性材料过氧化氢酶和谷胱甘肽为代表性改性添加剂,制备能够显著降低卷烟主流烟气关键有害成分含量的改性滤嘴和醋酸纤维丝束,并对其作用机理进行深入研究,主要研究内容及结果如下:
(1)设计并制作了一套单通道模拟吸烟及主流烟气收集装置;
(2)建立了使用薄层层析-毛细管气相色谱(TLC-GC/PID)同时定性、定量分析卷烟主流烟气中16种多环芳烃的测定方法;
(3)建立了使用高效液相色谱—串联质谱(HPLC-MS/MS)同时定性、定量分析主流烟气中四种烟草特有亚硝胺的测定方法;
(4)完善了以2-叔丁基硝酮(PBN)的苯溶液为自旋捕捉剂溶液,使用电子顺磁共振—电子自旋捕集技术(EPR-Spin trap)定性定量分析卷烟主流烟气中气相自由基含量的烟气自由基分析方法;
(5)系统研究了普通醋纤滤嘴生产工艺,确定在丝束的开松上胶过程中以干粉喷雾的方式将滤嘴改性添加剂加入滤嘴体系的改性方案;
(6)系统研究了烟用二醋酸纤维丝束的生产工艺,确定在丝束上油过程中,利用含添加剂上油乳液将可溶性改性添加剂均匀添加到烟用二醋酸纤维单丝表面的丝束改性方案;
(7)以固定化过氧化氢酶为代表性改性添加剂,制备了能够选择性降低卷烟主流烟气中气相自由基含量的改性滤嘴。结果表明,含有约10 mg/支固定化过氧化氢酶微粒的改性滤嘴中能使卷烟主流烟气中气相自由基含量下降38.84%,并且对卷烟常规指标(吸阻、水分和烟气烟碱量)没有明显影响;使用自制模拟吸烟装置,结合非常规模拟吸烟方案,发现主流烟气中总粒相物对改性滤嘴的降自由基作用具有明显促进作用;通过进一步试验证实改性滤嘴中的固定化过氧化氢酶主要是在有过氧化物存在的条件下,迅速催化降解主流烟气中的氮氧化物,破坏气相自由基反应链,从而有效降低卷烟主流烟气中气相自由基的含量;
(8)以还原型谷胱甘肽为代表性改性添加剂,制备了能够选择性降低卷烟主流烟气中气相自由基、烟草特有亚硝胺和酚类化合物含量的改性醋酸纤维丝束。结果表明,含添加剂油剂对醋酸纤维丝束的上油性能、机械加工性能及其物理性能没有显著影响;对改性醋纤丝束的应用实验结果表明,改性醋纤表面的还原型谷胱甘肽有效含量低至0.32 mg/g(换算至滤嘴添加量约0.032 mg/支)时,仍然能显著降低卷烟主流烟气中气相自由基、烟草特有亚硝胺和酚类物质含量,其中气相自由基含量同比下降25.07%,TSNAs总量同比下降21.52%(其中NNK和NNN的降幅分别达到32.14%和28.04%),总酚同比下降16.54%;并且由于改性醋纤滤嘴能有效降低烟气中的酚类化合物,因而降低了烟气的刺激性,使卷烟烟气口感更加柔和;
(9)分别使用MDA-TBA方法和Ames波动试验,从卷烟主流烟气的氧化损伤作用和烟气冷凝物致突变性,评价改性滤嘴对卷烟安全性的影响。结果表明,含有活性添加剂的改性醋纤滤嘴能够显著降低卷烟烟气的生物毒性,通过在滤嘴中添加固定化过氧化氢酶或谷胱甘肽对主流烟气及其气相成分的急性氧化损伤作用的抑制效率分别为43.57%、41.65%和38.57%、39.32%;使主流烟气直接和间接致突变能力分别下降41.94%和11.54%和32.35%和55.56%。
综上所述,本文以过氧化氢酶和谷胱甘肽为代表性生物活性材料,通过对卷烟滤嘴和烟用二醋酸纤维的相关改性,有效降低了卷烟主流烟气中气相自由基和烟草特有亚硝胺等关键有害成分的含量,提高了卷烟安全性。本研究对于开发低危害卷烟产品具有重要应用价值,对于烟用醋酸纤维丝束及滤嘴的进一步改性和提高具有较好的理论和实践意义。
Health risks induced by smoking are getting worse and worse. International tobacco industries have thus begun to search for the less harmful cigarettes. In the present thesis, systematic study was conducted on the production procedures of cellulose fiber and the corresponding filter tips. Catalase and glutathione have been studied as the representative bioactive materials. Modified cellulose fiber capable of reducing the yields of key toxicants in mainstream cigarette smoke was produced and, detailed investigations have been focused on the related mechanisms. Main research contents and results are as follows:
(1) A single-port smoking device was designed and prepared for smoking simulation and sampling followed by smoke analysis.
(2) Thin-layer chromatography (TLC) coupled with capillary gas chromatography was adopted for the simultaneously qualitative and quantitative analysis of 16 kinds polycyclic aromatic hydrocarbons (PAHs) including benzo[a]pyrene in mainstream cigarette smoke.
(3) High performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS) was adopted for the simultaneous determination of four kinds of tobacco specific nitrosamines (TSNAs) in mainstream cigarette smoke.
(4) Electronic paramagnetic resonance coupled with spin trapping method (EPR-Spin trap) was improved for the determination of free radicals in vapor-phase mainstream cigarette smoke.
(5) Systematic research was conducted on the production process of acetate filter rod from cellulose acetate fibers. It was concluded from the results of the researching test that non-soluble additives for the modification of filter tip could be homogeneously sprayed into the matrix of opened cellulose acetate fibers during the procedure of rubberizing.
(6) Systematic research was conducted on the production process of cellulose acetate fibers from the raw pulp. It was concluded from the result of the researching test that soluble additives could be homogeneously attached onto the surface of fiber through oiling process.
(7) A kind of modified filter tip capable of selectively reducing vapor-phase free radicals was produced with the CTS-CAT as additive. Is was found that the modified filter containing 10 mg CTS-CAT microparticle could apparently lower the level of vapor-phase free radicals by 38.84|%, without affecting the nature of cigarette smoke such as draw resistance, moisture and the nicotine delivery. Judging from the results of the nitrogen oxides assay corresponding to the unconventional smoking simulations, it could be concluded that the selective reduction of vapor-phase free radicals by the modified filter tip could be attributed to the catalase-dependent breakdown of nitric oxide in the presence of peroxides (most probably being H_2O_2), which could have been derived from the dismutation of TPM.
(8) A kind of modified cellulose acetate fiber capable of selective reducing key toxicants including vapor-phase free radicals, TSNAs and phenolic compounds was produced with GSH as the additive. Is was found that the GSH content on the surface of the modified fiber was as low as 0.32 mg/g (approximately 0.032 mg for a single filter tip), while showed promising ability to lower the level of vapor-phase free radicals, TSNAs and phenolic compounds by 25.07%, 21.52%(32.14% and 28.04% for NNN and NNK respectively) and 16.54 %, respectively. It was further found by professional sensory evaluation that the GSH containing filter may improve the cigarette smoke flavor due mostly to the reduction of biting-taste compounds such as phenolic compounds..
(9) MDA-TBA assay and Ames fluctuation test were adopted to evaluate the impact of modification on the oxidative damage of mainstream cigarette smoke as well as the mutagenic and mutagencity of the corresponding condensate. As a result, the immobilized catalase added in filter apparently reduced the oxidative damage of mainstream smoke and its vapor phase components by 43.57 % and 41.65%, 38.57% and 39.32% GSH, respectively. Beside, the direct and indirect mutagencity potency was reduced by 41.94%, 11.54% by the immobilized catalase, and 32.35%, 55.56 % by GSH, respectively.
In conclusion, catalase and glutathione were studied as the representative bioactive materials for the reduction of key toxicants in mainstream cigarette smoke. As a result, smoking-related hazardous was alleviated to some extend. The study in this thesis was not only supposed to have practical significance and application importance for the development of less harmful tobacco products, but also of significance for the further improvement of acetate fiber and cigarette filter tips.
(1)设计并制作了一套单通道模拟吸烟及主流烟气收集装置;
(2)建立了使用薄层层析-毛细管气相色谱(TLC-GC/PID)同时定性、定量分析卷烟主流烟气中16种多环芳烃的测定方法;
(3)建立了使用高效液相色谱—串联质谱(HPLC-MS/MS)同时定性、定量分析主流烟气中四种烟草特有亚硝胺的测定方法;
(4)完善了以2-叔丁基硝酮(PBN)的苯溶液为自旋捕捉剂溶液,使用电子顺磁共振—电子自旋捕集技术(EPR-Spin trap)定性定量分析卷烟主流烟气中气相自由基含量的烟气自由基分析方法;
(5)系统研究了普通醋纤滤嘴生产工艺,确定在丝束的开松上胶过程中以干粉喷雾的方式将滤嘴改性添加剂加入滤嘴体系的改性方案;
(6)系统研究了烟用二醋酸纤维丝束的生产工艺,确定在丝束上油过程中,利用含添加剂上油乳液将可溶性改性添加剂均匀添加到烟用二醋酸纤维单丝表面的丝束改性方案;
(7)以固定化过氧化氢酶为代表性改性添加剂,制备了能够选择性降低卷烟主流烟气中气相自由基含量的改性滤嘴。结果表明,含有约10 mg/支固定化过氧化氢酶微粒的改性滤嘴中能使卷烟主流烟气中气相自由基含量下降38.84%,并且对卷烟常规指标(吸阻、水分和烟气烟碱量)没有明显影响;使用自制模拟吸烟装置,结合非常规模拟吸烟方案,发现主流烟气中总粒相物对改性滤嘴的降自由基作用具有明显促进作用;通过进一步试验证实改性滤嘴中的固定化过氧化氢酶主要是在有过氧化物存在的条件下,迅速催化降解主流烟气中的氮氧化物,破坏气相自由基反应链,从而有效降低卷烟主流烟气中气相自由基的含量;
(8)以还原型谷胱甘肽为代表性改性添加剂,制备了能够选择性降低卷烟主流烟气中气相自由基、烟草特有亚硝胺和酚类化合物含量的改性醋酸纤维丝束。结果表明,含添加剂油剂对醋酸纤维丝束的上油性能、机械加工性能及其物理性能没有显著影响;对改性醋纤丝束的应用实验结果表明,改性醋纤表面的还原型谷胱甘肽有效含量低至0.32 mg/g(换算至滤嘴添加量约0.032 mg/支)时,仍然能显著降低卷烟主流烟气中气相自由基、烟草特有亚硝胺和酚类物质含量,其中气相自由基含量同比下降25.07%,TSNAs总量同比下降21.52%(其中NNK和NNN的降幅分别达到32.14%和28.04%),总酚同比下降16.54%;并且由于改性醋纤滤嘴能有效降低烟气中的酚类化合物,因而降低了烟气的刺激性,使卷烟烟气口感更加柔和;
(9)分别使用MDA-TBA方法和Ames波动试验,从卷烟主流烟气的氧化损伤作用和烟气冷凝物致突变性,评价改性滤嘴对卷烟安全性的影响。结果表明,含有活性添加剂的改性醋纤滤嘴能够显著降低卷烟烟气的生物毒性,通过在滤嘴中添加固定化过氧化氢酶或谷胱甘肽对主流烟气及其气相成分的急性氧化损伤作用的抑制效率分别为43.57%、41.65%和38.57%、39.32%;使主流烟气直接和间接致突变能力分别下降41.94%和11.54%和32.35%和55.56%。
综上所述,本文以过氧化氢酶和谷胱甘肽为代表性生物活性材料,通过对卷烟滤嘴和烟用二醋酸纤维的相关改性,有效降低了卷烟主流烟气中气相自由基和烟草特有亚硝胺等关键有害成分的含量,提高了卷烟安全性。本研究对于开发低危害卷烟产品具有重要应用价值,对于烟用醋酸纤维丝束及滤嘴的进一步改性和提高具有较好的理论和实践意义。
Health risks induced by smoking are getting worse and worse. International tobacco industries have thus begun to search for the less harmful cigarettes. In the present thesis, systematic study was conducted on the production procedures of cellulose fiber and the corresponding filter tips. Catalase and glutathione have been studied as the representative bioactive materials. Modified cellulose fiber capable of reducing the yields of key toxicants in mainstream cigarette smoke was produced and, detailed investigations have been focused on the related mechanisms. Main research contents and results are as follows:
(1) A single-port smoking device was designed and prepared for smoking simulation and sampling followed by smoke analysis.
(2) Thin-layer chromatography (TLC) coupled with capillary gas chromatography was adopted for the simultaneously qualitative and quantitative analysis of 16 kinds polycyclic aromatic hydrocarbons (PAHs) including benzo[a]pyrene in mainstream cigarette smoke.
(3) High performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS) was adopted for the simultaneous determination of four kinds of tobacco specific nitrosamines (TSNAs) in mainstream cigarette smoke.
(4) Electronic paramagnetic resonance coupled with spin trapping method (EPR-Spin trap) was improved for the determination of free radicals in vapor-phase mainstream cigarette smoke.
(5) Systematic research was conducted on the production process of acetate filter rod from cellulose acetate fibers. It was concluded from the results of the researching test that non-soluble additives for the modification of filter tip could be homogeneously sprayed into the matrix of opened cellulose acetate fibers during the procedure of rubberizing.
(6) Systematic research was conducted on the production process of cellulose acetate fibers from the raw pulp. It was concluded from the result of the researching test that soluble additives could be homogeneously attached onto the surface of fiber through oiling process.
(7) A kind of modified filter tip capable of selectively reducing vapor-phase free radicals was produced with the CTS-CAT as additive. Is was found that the modified filter containing 10 mg CTS-CAT microparticle could apparently lower the level of vapor-phase free radicals by 38.84|%, without affecting the nature of cigarette smoke such as draw resistance, moisture and the nicotine delivery. Judging from the results of the nitrogen oxides assay corresponding to the unconventional smoking simulations, it could be concluded that the selective reduction of vapor-phase free radicals by the modified filter tip could be attributed to the catalase-dependent breakdown of nitric oxide in the presence of peroxides (most probably being H_2O_2), which could have been derived from the dismutation of TPM.
(8) A kind of modified cellulose acetate fiber capable of selective reducing key toxicants including vapor-phase free radicals, TSNAs and phenolic compounds was produced with GSH as the additive. Is was found that the GSH content on the surface of the modified fiber was as low as 0.32 mg/g (approximately 0.032 mg for a single filter tip), while showed promising ability to lower the level of vapor-phase free radicals, TSNAs and phenolic compounds by 25.07%, 21.52%(32.14% and 28.04% for NNN and NNK respectively) and 16.54 %, respectively. It was further found by professional sensory evaluation that the GSH containing filter may improve the cigarette smoke flavor due mostly to the reduction of biting-taste compounds such as phenolic compounds..
(9) MDA-TBA assay and Ames fluctuation test were adopted to evaluate the impact of modification on the oxidative damage of mainstream cigarette smoke as well as the mutagenic and mutagencity of the corresponding condensate. As a result, the immobilized catalase added in filter apparently reduced the oxidative damage of mainstream smoke and its vapor phase components by 43.57 % and 41.65%, 38.57% and 39.32% GSH, respectively. Beside, the direct and indirect mutagencity potency was reduced by 41.94%, 11.54% by the immobilized catalase, and 32.35%, 55.56 % by GSH, respectively.
In conclusion, catalase and glutathione were studied as the representative bioactive materials for the reduction of key toxicants in mainstream cigarette smoke. As a result, smoking-related hazardous was alleviated to some extend. The study in this thesis was not only supposed to have practical significance and application importance for the development of less harmful tobacco products, but also of significance for the further improvement of acetate fiber and cigarette filter tips.
引文
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