CN115226729A - 用于降低雪茄烟有害成分的烟叶叶面喷施物、方法及烟丝 - Google Patents

用于降低雪茄烟有害成分的烟叶叶面喷施物、方法及烟丝 Download PDF

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CN115226729A
CN115226729A CN202211040095.0A CN202211040095A CN115226729A CN 115226729 A CN115226729 A CN 115226729A CN 202211040095 A CN202211040095 A CN 202211040095A CN 115226729 A CN115226729 A CN 115226729A
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tobacco
foliar spray
spray
leaf
leaf extract
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CN115226729B (zh
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杨春雷
杨锦鹏
周骏
杨美
杨勇
余君
李宗平
饶雄飞
覃光炯
乔保明
白若石
马雁军
邓显豹
秦文章
樊凯孝
邓灿东
魏永乐
樊友伦
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Tobacco Research Institute of Hubei Province
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Abstract

本申请公开了用于降低雪茄烟有害成分的烟叶叶面喷施物、降低烟叶烤制后致癌成分的方法及烤烟烟丝。该叶面喷施物包含荷叶提取物，所述有害化学成分包括N‑亚硝基降烟碱、4‑(N‑甲基‑亚硝胺)‑1‑(3‑吡啶基)‑1‑丁酮、N‑亚硝基假木贼碱和N‑亚硝基新烟碱。采用荷叶提取物在烟叶采收调制前的大田进行叶面喷施；不仅显著提升烟草农艺性状，经济形状，缓解了烟叶褐变，还有效降低了烟叶特有亚硝胺和烟碱等有害化学成分。

Description

用于降低雪茄烟有害成分的烟叶叶面喷施物、方法及烟丝
技术领域
本申请涉及雪茄烟有害成分降低技术领域，尤其涉及用于降低雪茄烟有害成分的烟叶叶面喷施物、方法及烟丝。
背景技术
烟叶质量受土壤、气候、品种、栽培技术、烘烤技术等诸多因素的影响。烤制后的卷烟中一般含有特有亚硝胺(TSNAs)因其强列的致癌作用而备受关注。如何在既改善卷烟吸食品质的同时又能从复杂的卷烟主流烟气中降低有害成分含量是烟草科研工作者所面临的一项挑战性课题。据相关研究结果显示，在烟叶中添加一些能够阻断亚硝化反应的黄酮类物质，不仅能够有效降低烟叶中亚硝胺成分含量，还具有清咽、润喉、镇咳等功能，并可以达到在卷烟中添加这类中草药既能发挥保健功效同时又能降低烟草有害成分的双重目的。例如，根据中药药典记载，葛根、陈皮、菊花等中草药具有较好的清咽、润喉、镇咳等作用，能否提取到这些中草药的有效成分。然而，这些中药成分复杂，并不能明确具体成分的作用，而且降低亚硝胺作用有效，采用这些名贵中药材成本高昂，对于烤制卷烟的生产成本和产量受限。
发明内容
有鉴于此，本申请的目的在于开发一种开发一种能够有效降低烤制卷烟中亚硝胺成分的方法及产品，并有效降低烤制卷烟的生产成本。
第一方面，本申请实施例公开了用于降低雪茄烟有害成分的烟叶叶面喷施物，所述叶面喷施物包含荷叶提取物，所述有害化学成分包括N-亚硝基降烟碱、4-(N-甲基-亚硝胺)-1-(3-吡啶基)-1-丁酮、N-亚硝基假木贼碱和N-亚硝基新烟碱。
在本申请实施例中，所述荷叶提取物采用如下方法制得：
将新鲜采摘的荷叶与清水按质量比1:3于组织捣碎机中捣碎，然后将捣碎后的混合悬浮液注入循环超声提取机进行超声提取，超声提取操作参数为提取温度为室温25±5℃、超声功率1000W、转速1200rpm、提取时间(循环时长)30±5min；超声后过滤，取滤液稀释即得。
在本申请实施例中，所述荷叶提取物包括松脂醇、表松脂醇、异落叶松树脂醇、牛蒡酚F、黑麦草内酯、黄柏酮、4,5-二氢布卢门醇A、3-氧-α-紫罗兰醇和去氢催吐萝芙叶醇中的至少一种。
在本申请实施例中，所述烟叶叶面喷施物按重量百分比计包含0.001～0.04％异落叶松树脂醇、0.001～0.03％黑麦草内酯、0.001～0.04％黄柏酮。
在本申请实施例中，所述烟叶叶面喷施物按重量百分比计包含0.001～0.04％4,5-二氢布卢门醇A、0.001～0.04％3-氧-α-紫罗兰醇和0.001～0.03％去氢催吐萝芙叶醇。
在本申请实施例中，所述烟叶叶面喷施物按重量百分比计包含0.001～0.02％异落叶松树脂醇、0.001～0.01％黑麦草内酯、0.001～0.015％黄柏酮、0.001～0.02％4,5-二氢布卢门醇A、0.001～0.025％3-氧-α-紫罗兰醇和0.001～0.025％去氢催吐萝芙叶醇。
在本申请实施例中，所述烟叶叶面喷施物按重量百分比计包含0.001～0.015％松脂醇、0.001～0.01％表松脂醇、0.001～0.015％异落叶松树脂醇、0.001～0.01％黑麦草内酯、0.001～0.015％黄柏酮、0.001～0.02％4,5-二氢布卢门醇A、0.001～0.015％3-氧-α-紫罗兰醇和0.001～0.015％去氢催吐萝芙叶醇。
在本申请实施例中，所述烟叶叶面喷施物还包括用于溶解所述荷叶提取物的试剂以及其他农作物喷施可用的辅料。
第二方面，本申请实施例公开了利用第一方面所述烟叶叶面喷施物喷施制得的烤烟烟丝，其中包含不高于556.95ng/gNNN、4.15ng/gNNK、299.52ng/gNAB和65.78ng/gNAT。
第三方面，本申请实施例公开了降低烟叶烤制后致癌成分的方法，包括在烟叶成熟期采收前5天喷施第一方面所述的烟叶叶面喷施物的步骤。
与现有技术相比，本申请至少具有以下有益效果：
本申请实实施例公开了采用荷叶提取物在烟叶采收调制前的大田进行三次叶面喷施；不仅显著提升烟草农艺性状，经济形状，缓解了烟叶褐变，还有效降低了烟叶特有亚硝胺和烟碱等有害化学成分，增加了烟叶香气成含量，从而综合提升了烟草产值常量和品质；还降低烤制卷烟的生产成本，实现了对荷叶资源的回收利用。
具体实施方式
为了使本申请的目的、技术方案及优点更加清楚明白，以下结合实施例对本申请进行进一步详细说明。应当理解，此处所描述的具体实施例仅仅用以解释本申请，并不用于限定本申请。
一、材料与方法
1、试验品种
试验于湖北省烟草科学研究院进行，试验材料为烤烟(Nicotiana tobacum L.，楚雪14号)
2、育苗及土培期
将烟草种子分别播种于烟草育苗盘(浙江博成新材料有限公司)内，基质包含重量比为2:1的泥炭和蛙石中，每孔一粒，采用漂浮育苗方式在温室内育苗，光照为自然光源，播种30天后移栽至填充有培养土的培养盆内，土壤包含稻田土和基质的混合重量比为5:2，置于田间环境生长30天。
3、团棵期、旺长期、打顶期及成熟期
待烟草植株生长进入团棵期，烟株自下向上第12片烟叶长至叶长为12cm时记为0d，然后分别将烟株移入智能人工气候室(江西省进贤绿佳温室工程有限公司)，控温温度为18～25℃下培养60d。
光周期(昼14h:06:00-20:00夜10h:20:00-06:00)，昼夜相对湿度65/55％，CO2浓度为450mmol。根据一天中温度变化的趋势，设置在18～25℃范围内的动态的温度变化，每隔一天各个处理下烟株分别浇水1kg/株，其它管理与大田管理一致。
在团稞期、旺长期、现蕾期、打顶期及成熟期的各生长期间，可施用普通有机叶面肥(购自萍乡市红土地腐植酸有限公司)。
烟草生长进入成熟期后，常规采收晾制，烤制后，即可得到卷烟原料。
4、叶面喷施液
本实验主要研究本申请实施例提供的叶面喷施液对卷烟原料的质量，烟草的农业性状以及发病情况的影响。将在智能人工气候室中培育的烟草植株分为10组，分别采用不同的供试品溶液进行喷施。
本申请实施例提供的供试品溶液包含荷叶提取物，该荷叶提取物采用如下方法制得：
将新鲜采摘的荷叶与清水按质量比1:3于组织捣碎机中捣碎，然后将捣碎后的混合悬浮液注入循环超声提取机进行超声提取，超声提取操作参数为提取温度为室温25±5℃、超声功率1000W、转速1200rpm、提取时间(循环时长)30±5min；超声后过滤，取滤液稀释12倍即为作为供试品1。
经过相关文献探索(荷叶中化学成分的分离与鉴定[J]，沈阳药科大学学报，2012年7月；荷叶中化学成分的分离与鉴定(III)[J]，天津中医药大学学报，2015年4月；荷叶中化学成分的分离与鉴定(II)[J]，沈阳药科大学学报，2014年5月)发现，荷叶提取物的主要成分包括松脂醇(上海泰坦科技股份有限公司，CAS:8006-39-1，纯度：95％+(GC))、表松脂醇(上海泰坦科技股份有限公司，纯度：95％+(GC))、异落叶松树脂醇(云南西力生物技术股份有限公司，CAS:548-29-8纯度：97.5％)、黄芩新素Ⅱ(上海源叶生物科技有限公司，CAS:55084-08-7纯度：HPLC≥98％)、粘毛黄芩素Ⅲ(云南西力生物技术股份有限公司，CAS:92519-91-0，纯度：98.0％)、牛蒡酚F(成都彼样生物科技有限公司，CAS:69394-17-8，纯度：HPLC≥98％)、黑麦草内酯(四川省维克奇生物科技有限公司，CAS:5989-02-6，纯度：HPLC≥98％)、黄柏酮(化夏化学，CAS:751-03-1纯度：99％HPLC)、4,5-二氢布卢门醇A(源叶生物，CAS号：155418-97-6,HPLC≥96％)，3-氧-α-紫罗兰醇(四川百奥克睿生物科技有限公司，CAS：896107-70-3，纯度：98％HPLC)和去氢催吐萝芙叶醇(源叶生物，CAS39763-33-2)。
为此，本实验将利用上述的荷叶提取物和竹叶提取物对烟草植株进行喷施，以探究其对烟草植株以及成烟后质量的影响。
表1
Figure BDA0003820623950000051
Figure BDA0003820623950000061
为此，本实验采用上述供试品溶液对烟草种植过程中进行喷施，具体喷施情况如表2所示。
表2
实施方式 喷施方案
实施例1 供试品1于成熟期采收前5天喷施
实施例2 供试品2于成熟期采收前5天喷施
实施例3 供试品3于成熟期采收前5天喷施
实施例4 供试品4于成熟期采收前5天喷施
实施例5 供试品5于成熟期采收前5天喷施
实施例6 供试品6于成熟期采收前5天喷施
实施例7 供试品7于成熟期采收前5天喷施
对比例1 未喷施上述任何供试品1～8
对比例2 供试品3于团稞期喷施
对比例3 供试品3于旺长期喷施
对比例4 供试品3于打顶期喷施
对比例5 供试品8于成熟期采收前5天喷施
5、烟株农艺性状调查
经过上述实施例1～7和对比例1～5的每一处理组别中随机选择10株烟株挂牌标记，按标准YC/T 142-1998《烟草农艺性状调查方法》，定点定株调查烟草团棵期、打顶期和成熟期农艺性状。其主要包括株高、茎围、烟株的有效叶面积。叶面积(cm2)＝0.6345×叶长(cm)×叶宽(cm)。
6、烟草青枯病发病情况调查
针对田间病害发生特点，对烟草青枯病进行了调查，根据GB/23222-2008《烟草病虫害调查与分级》分级国家标准，采用5点取样方法，每点固定调查10株，每3d调查一次。并根据公式计算发病率、病情指数和相对防效。
发病率(％)＝(发病株数/调查总株数)×100％
病情指数＝[(∑(发病株数)×病级代表数)/(调查总株数×病级最高值)]×100％
病情调查参照GB/23222-2008《烟草病虫害分级及调查方法》(以株为单位)：0级:全株无病；1级:茎部偶有褪绿斑，或病侧二分之一以下叶片凋萎；3级:茎部有黑色条斑，但不超过茎高二分之一，或病侧二分之一至三分之二叶片凋萎；5级:茎部黑色条斑超过茎高二分之一，但未到达茎顶部，或病侧三分之二以上叶片凋萎；7级:茎部黑色条斑到达茎顶部，或病株叶片全部凋萎；9级:病株基本枯死。
7、烤烟褐变性状
取烘干粉碎后的烟叶5g置于500mL烧杯中，加入250mL蒸馏水，于超声波清洗15min后，过滤，取滤液作为样品储备液。测定时将样品储备液按适当倍数进行稀释，紫外分光光度计于D420nm条件下测定其吸光值，即为褐变度。
8、烤烟TSNAs含量
采用如下方法检测烤烟烟丝中的TSNAs含量，包括N-亚硝基降烟碱、4-(N-甲基-亚硝胺)-1-(3-吡啶基)-1-丁酮、N-亚硝基假木贼碱和N-亚硝基新烟碱的含量。
(1)样品预处理：
将上述实施例1～7和对比例1～5分别制得的烤烟，粉粹后，加入至PBS缓冲液中，配制成25mg/mL溶液，超声处理1h，冷却后，于5000rpm离心5min，取上清，上样至硅藻土层析柱，静置吸附30min后，用二氯甲烷洗脱，收集洗脱液，加入无水硫酸钠干燥涡阳，于浓缩仪器中于35℃氮气氛围浓缩至1mL。
(2)标准品：N-亚硝基降烟碱(简称为NNN，北京百灵威科技有限公司，CAS:150824-47-8纯度：95％)、4-(N-甲基-亚硝胺)-1-(3-吡啶基)-1-丁酮(NNK，武汉丰泰威远科技有限公司，CAS号64091-91-4纯度：95％)、N-亚硝基假木贼碱(NAB，凯试(上海)科技有限公司，CAS:1133-64-8纯度：98％)、N-亚硝基新烟碱(NAT北京百灵威科技有限公司，CAS:887407-16-1纯度：98％)
(3)毛细管气相色谱法检测：采用配有NPD的安捷伦气相色谱仪6890测定，毛细管柱为db-5,30m×0.32mm×1μm,进样温度为250℃，检测其温度为280℃，升温程序：初温50℃保持4min；5℃/min升至200℃，保持4min，20℃/min升至260℃，保持20min；载气流速为3mL/min,辅助气流速度为20mL/min，氢气流速为3mL/min，空气流速为120mL/min，分流比为20:1，进样量为1μL。
9、烤后烟叶感官质量
参照《YC/T 530-2015烤烟烟叶质量风格特色感官评价方法》进行评价。
10、数据处理
所有测试数据均以平均值和标准偏差表示，应用SPSS13.0软件处理数据，并对每列数据进行多重比较和显著性差异标记。
二、结果
表3
Figure BDA0003820623950000081
Figure BDA0003820623950000091
由表2呈现了经过实施例1～7和对比例1～5提供的供试品溶液对烟草植株进行叶面喷施后的株高、叶面积以及茎围结果。由表2可知，相对于未进行叶面喷施的对比例1，实施例1～7和对比例2～5的株高、叶面积以及茎围均有显著提升，由此说明，经过本申请实施例提供的荷叶提取物、竹叶提取物以及其中的相关活性成分进行叶面喷施，有利于烟草作物的生长。
表4
实施方式 发病率 病情指数
实施例1 23.56±0.32b 8.18±0.13b
实施例2 23.62±0.24b 8.22±0.09b
实施例3 21.72±0.34c 8.05±0.08c
实施例4 21.75±0.43c 8.02±0.03c
实施例5 21.89±0.32c 8.12±0.11bc
实施例6 21.82±0.54c 8.09±0.07c
实施例7 20.78±0.13d 7.92±0.08d
对比例1 25.08±0.27a 8.37±0.05a
对比例2 21.73±0.17c 8.07±0.07c
对比例3 21.71±0.08c 8.05±0.12c
对比例4 21.68±0.24c 8.03±0.06c
对比例5 21.70±0.16c 8.04±0.07c
由表4可知，相对于未进行叶面喷施的对比例1，实施例1～7和对比例2～5的发病率和病情指数均有显著降低，由此说明，经过本申请实施例提供的荷叶提取物、竹叶提取物以及其中的相关活性成分进行叶面喷施，有利于减少烟草作物病虫害发生。
表5
Figure BDA0003820623950000092
Figure BDA0003820623950000101
表5呈现了了各实施例1～7和对比例1～5对烟草进行叶面喷施供试品溶液后，烤制得到卷烟原料在20℃、25℃和30℃温度条件下陈化5个月后的褐变度结果。由表5可知，相对于未进行叶面喷施的对比例1，实施例1～7制得的烤烟烟丝在20℃、25℃和30℃温度条件下陈化5个月后的褐变度均显著降低，由此说明，经过本申请实施例提供的荷叶提取物、竹叶提取物以及其中的相关活性成分进行叶面喷施，有利降低烤烟陈化过程的褐变。
另外，结合表3、4的结果发现，对比例2～5提供的叶面喷施方案虽然有利于提升烟株的农艺性状，但是其不利于减少烤制后的卷烟原料在不同温度下的降低陈化褐变发生。
表6
Figure BDA0003820623950000102
Figure BDA0003820623950000111
表6呈现了了各实施例1～7和对比例1～5对烟草进行叶面喷施供试品溶液后，烤制得到卷烟原料中NNN、NNK、NAB和NAT含量。由表6可知，相对于未进行叶面喷施的对比例1，实施例1～7制得的烤烟烟丝中NNN、NNK、NAB和NAT含量均显著低于对比例。由此说明，经过本申请实施例提供的荷叶提取物、竹叶提取物以及其中的相关活性成分进行成熟期叶面喷施，有利降低烤烟中相关的TSNAs含量，为制得低致癌性卷烟提供帮助。
由此，经过上述实施例提供的方法，本申请实施例还公开了一种烤烟烟丝，其中包含不高于556.95ng/gNNN、4.15ng/gNNK、299.52ng/gNAB和65.78ng/gNAT。
为此，本申请还对实施例1制得的烤烟烟丝和对比例1制得的烤烟烟丝进行感官质量评价，结果如表7所示，表7中为参照《烟草制品感官评价方法YC/T 415—2011执行标准》分析的实施例1和对比例1制得烟丝感官评分结果；其中，感官评价总分值为100分，香气质占比18分，香气量16分，杂气16分，刺激性20分，余味22分，燃烧性4分，灰色4分。
结果如表7所示，实施例1提供的烤烟烟丝的感官评吸质量较好，香气质、香气量有所改善；而对比例1提供的烤烟烟丝感官评吸质量最差，杂气增多，刺激性增强，余味变差。
表7
Figure BDA0003820623950000112
Figure BDA0003820623950000121
综上所述，本申请实实施例公开了采用荷叶提取物和/或竹叶提取物在烟叶采收调制前的大田进行三次叶面喷施；不仅显著提升烟草农艺性状，经济形状，缓解了烟叶褐变，还有效降低了烟叶特有亚硝胺和烟碱等有害化学成分，增加了烟叶香气成含量，从而综合提升了烟草产值常量和品质。
以上所述，仅为本申请较佳的具体实施方式，但本申请的保护范围并不局限于此，任何熟悉本技术领域的技术人员在本申请揭露的技术范围内，可轻易想到的变化或替换，都应涵盖在本申请的保护范围之内。

Claims (10)

1.用于降低雪茄烟有害成分的烟叶叶面喷施物，所述叶面喷施物包含荷叶提取物，所述有害化学成分包括N-亚硝基降烟碱、4-(N-甲基-亚硝胺)-1-(3-吡啶基)-1-丁酮、N-亚硝基假木贼碱和N-亚硝基新烟碱。
2.根据权利要求1所述的烟叶叶面喷施物，其特征在于，所述荷叶提取物采用如下方法制得：
将新鲜采摘的荷叶与清水按质量比1:3于组织捣碎机中捣碎，然后将捣碎后的混合悬浮液注入循环超声提取机进行超声提取，超声提取操作参数为提取温度为室温25±5℃、超声功率1000W、转速1200rpm、提取时间(循环时长)30±5min；超声后过滤，取滤液稀释即得。
3.根据权利要求1所述的烟叶叶面喷施物，其特征在于，所述荷叶提取物包括松脂醇、表松脂醇、异落叶松树脂醇、牛蒡酚F、黑麦草内酯、黄柏酮、4,5-二氢布卢门醇A、3-氧-α-紫罗兰醇和去氢催吐萝芙叶醇中的至少一种。
4.根据权利要求1所述的烟叶叶面喷施物，其特征在于，所述烟叶叶面喷施物按重量百分比计包含0.001～0.04％异落叶松树脂醇、0.001～0.03％黑麦草内酯、0.001～0.04％黄柏酮。
5.根据权利要求1所述的烟叶叶面喷施物，其特征在于，所述烟叶叶面喷施物按重量百分比计包含0.001～0.04％4,5-二氢布卢门醇A、0.001～0.04％3-氧-α-紫罗兰醇和0.001～0.03％去氢催吐萝芙叶醇。
6.根据权利要求1所述的烟叶叶面喷施物，其特征在于，所述烟叶叶面喷施物按重量百分比计包含0.001～0.02％异落叶松树脂醇、0.001～0.01％黑麦草内酯、0.001～0.015％黄柏酮、0.001～0.02％4,5-二氢布卢门醇A、0.001～0.025％3-氧-α-紫罗兰醇和0.001～0.025％去氢催吐萝芙叶醇。
7.根据权利要求1所述的烟叶叶面喷施物，其特征在于，所述烟叶叶面喷施物按重量百分比计包含0.001～0.015％松脂醇、0.001～0.01％表松脂醇、0.001～0.015％异落叶松树脂醇、0.001～0.01％黑麦草内酯、0.001～0.015％黄柏酮、0.001～0.02％4,5-二氢布卢门醇A、0.001～0.015％3-氧-α-紫罗兰醇和0.001～0.015％去氢催吐萝芙叶醇。
8.根据权利要求1～7任一项所述的烟叶叶面喷施物，其特征在于，所述烟叶叶面喷施物还包括用于溶解所述荷叶提取物的试剂以及其他农作物喷施可用的辅料。
9.利用权利要求1～7任一项所述的烟叶叶面喷施物喷施制得的烤烟烟丝，其中包含不高于556.95ng/gNNN、4.15ng/gNNK、299.52ng/gNAB和65.78ng/gNAT。
10.降低烟叶烤制后致癌成分的方法，包括在烟叶成熟期采收前5天喷施如权利要求1～7任一项所述的烟叶叶面喷施物的步骤。
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