CN105969834A - 一种玳玳花烟用香料的制备方法 - Google Patents
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Abstract
本发明是一种玳玳花烟用香料的制备方法。该香料是由以下方法制备而得:A.发酵菌种活化,根霉活化;B.玳玳花发酵原料预处理;C.玳玳花发酵;D.发酵后玳玳花挥发油提取,即得所述的玳玳花烟用香料。本发明的香料对卷烟的香气和吸味品质改善作用明显,香气逼真,强度高,与香烟协调。本发明的方法工艺过程简单易行,设备简单,可直接用于卷烟加香加料,生产的产品可溶性好,便于推广应用。本发明所用的原料来源广泛,成本低,价格低廉,经济效益好。
Description
技术领域
本发明涉及烟用香料制备领域,特别是涉及到一种玳玳花烟用香料的制备方法。
背景技术
玳玳花(Citrus aurantium),别名回青橙、枳壳花、酸橙花,常绿灌木,枝楞、细长,叶互生,革质,椭圆形,春夏4-5月开白花,香气浓郁,果实扁球形,当年冬季为橙红色,翌年夏季又变青,故称“回青橙”,因有果实数代同生一树习性,亦称“公孙桔”。主要栽培在浙江、江苏、福建和四川等地,玳玳为重要栽培芳香植物,其花、枝叶和果皮的精油都是重要的调香原料,广泛用于食品、化妆品、香皂等香精中;特别是花精油,其香气优雅,主要用于配制高档香精。
目前,利用微生物发酵技术制备烟用香料,是一门全新的技术领域。微生物在增殖过程中可产生庞大的高活性酶系,如多糖水解酶类、蛋白酶类、纤维素酶类、酯化酶类、氧化还原酶类及裂合酶类等,在酶促作用、化学作用及微生物体内复杂代谢的协同作用下,使原料发生分解、降解、氧化、还原、聚合、偶联、转化等作用,形成复杂的低分子化合物,其中包括各种香味化合物,如醇类、醛类、酮类、酸类、酯类、酚类、呋喃类、吡嗪类、吡啶类和萜烯类等,这些香气物质无疑也是烟草的香气组分。但现有技术中,利用玳玳花为原料采用微生物发酵技术生产特色烟用香料的方法未见报道。
发明内容
本发明所要解决的技术问题在于提供一种能够采用微生物发酵技术生产玳玳花烟用香料的方法。
本发明一种玳玳花烟用香料的制备方法,其特征在于,按照如下步骤进行:
A.发酵菌种活化
①酵母菌种活化:选取微生物菌种,接种于准备好的YPD斜面培养基上,放置于25~30℃的恒温培养箱中培养24~48h使菌种活化,再将活化的菌种接种于YPD液体培养基中,于25~30℃恒温培养24~48h,得到活化的种子液;
②根霉活化:选取根霉菌种,接种于准备好的豆芽汁斜面培养基上,放置于25~30℃的恒温培养箱中培养48~72h使菌种活化,再将活化的菌种接种于YPD液体培养基中,于25~30℃恒温培养48~72h,得到活化的种子液;
B.玳玳花发酵原料预处理
选取清洁干燥的玳玳花作为原料,均匀喷洒玳玳花质量0.1~0.25倍的处理液,在115℃下灭菌5~15min,即得玳玳花原料;所述处理液含有质量分数1~5%葡萄糖,0.1~1%硫酸铵,0.1~0.5%七水硫酸镁,0.1~1%氯化钠,0.05~0.12%七水硫酸亚铁,余量为水;
C.玳玳花发酵
将活化酵母种子液和根霉分别按照1:10~20和1:5~10的质量比接种到无菌的玳玳花原料中,搅拌,使原料与种子液充分混合均匀,放置于25~32℃的培养箱中发酵2~7天;
D.发酵后玳玳花挥发油提取
取发酵后的玳玳花,用滤纸包成小包放入索氏提取器中,提取瓶内加入石油醚,55~60℃回流提取,提取至提取器中液体至无色,取出提取液,加无水硫酸钠低温干燥24h过滤,挥去滤液中的石油醚,即为所述的玳玳花烟用香料。
优选的,步骤A中所述的酵母菌种为酿酒酵母菌,根霉菌种为白地根霉。
优选的,步骤A中所述的YPD培养基为:葡萄糖20g,酵母膏10g,蛋白胨20g,蒸馏水100mL;如果选用斜面培养基则需加琼脂15g;配制完成后以115℃灭菌20min。
优选的,步骤A中所述的豆芽汁斜面培养基为:10%豆芽汁20ml,蔗糖5g,纯净水80ml,121℃灭菌20min。
优选的,所述步骤D中石油醚体积与发酵后的玳玳花的质量比为5~10g/mL。
微生物在增殖过程中可产生庞大的高活性酶系,如多糖水解酶类、蛋白酶类、纤维素酶类、酯化酶类、氧化还原酶类及裂合酶类等,在酶促作用、化学作用及微生物体内复杂代谢的协同作用下,以玳玳花为原料发生分解、降解、氧化、还原、聚合、偶联、转化等作用,形成复杂的低分子化合物,其中包括各种香味化合物,如醇类、醛类、酮类、酸类、酯类、酚类、呋喃类、吡嗪类、吡啶类和萜烯类等,这些香气物质无疑也是烟草的香气组分。
本发明的香料对卷烟的香气和吸味品质改善作用明显,香气逼真,强度高,与香烟协调。本发明的方法工艺过程简单易行,设备简单,可直接用于卷烟加香加料,生产的产品可溶性好,便于推广应用。本发明所用的原料来源广泛,成本低,价格低廉,经济效益好。
附图说明
图1为本发明一种玳玳花烟用香料的成分分析GC/MS图谱。
具体实施方式
为了更好的理解本发明的技术方案,下面结合实施例详细描述本发明提供的技术方案。下例仅用于进一步说明本发明,但不应理解为对本发明的限制。
一种玳玳花烟用香料的制备方法,按照如下步骤进行:
A.发酵菌种活化
①酵母菌种活化:选取微生物菌种,接种于准备好的YPD斜面培养基上,放置于25~30℃的恒温培养箱中培养24~48h使菌种活化,再将活化的菌种接种于YPD液体培养基中,于25~30℃恒温培养24~48h,得到活化的种子液;
②根霉活化:选取根霉菌种,接种于准备好的豆芽汁斜面培养基上,放置于25~30℃的恒温培养箱中培养48~72h使菌种活化,再将活化的菌种接种于YPD液体培养基中,于25~30℃恒温培养48~72h,得到活化的种子液;
B.玳玳花发酵原料预处理
选取清洁干燥的玳玳花作为原料,均匀喷洒玳玳花质量0.1~0.25倍的处理液,在115℃下灭菌5~15min,即得玳玳花原料;所述处理液含有质量分数1~5%葡萄糖,0.1~1%硫酸铵,0.1~0.5%七水硫酸镁,0.1~1%氯化钠,0.05~0.12%七水硫酸亚铁,余量为水;
C.玳玳花发酵
将活化酵母种子液和根霉分别按照1:10~20和1:5~10的质量比接种到无菌的玳玳花原料中,搅拌,使原料与种子液充分混合均匀,放置于25~32℃的培养箱中发酵2~7天;
D.发酵后玳玳花挥发油提取
取发酵后的玳玳花,用滤纸包成小包放入索氏提取器中,提取瓶内加入石油醚,55~60℃回流提取,提取至提取器中液体至无色,取出提取液,加无水硫酸钠低温干燥24h过滤,挥去滤液中的石油醚,即为所述的玳玳花烟用香料。
优选的,步骤A中所述的酵母菌种为酿酒酵母菌,根霉菌种为白地根霉。
优选的,步骤A中所述的YPD培养基为:葡萄糖20g,酵母膏10g,蛋白胨20g,蒸馏水100mL;如果选用斜面培养基则需加琼脂15g;配制完成后以115℃灭菌20min。
优选的,步骤A中所述的豆芽汁斜面培养基为:10%豆芽汁20ml,蔗糖5g,纯净水80ml,121℃灭菌20min。
优选的,所述步骤D中石油醚体积与发酵后的玳玳花的质量比为5~10g/mL。
实施例1
A.发酵菌种活化
①酵母菌活化:取酿酒酵母,接种于准备好的YPD斜面培养基上,放置于25℃的恒温培养箱中培养24h使菌种活化,再将活化的菌种接种于YPD液体培养基中,于25℃恒温培养24h,得到活化的种子液。YPD培养基:葡萄糖20g,酵母膏10g,蛋白胨20g,蒸馏水100mL(斜面培养基需加琼脂15g),以115℃灭菌20min。
②根霉活化:选取根霉菌种,接种于准备好的豆芽汁斜面培养基上,放置于25~30℃的恒温培养箱中培养48~72h使菌种活化,再将活化的菌种接种于YPD液体培养基中,于25~30℃恒温培养48~72h,得到活化的种子液;
B.玳玳花发酵原料预处理
选取200g玳玳花作为原料,用喉头喷雾器均匀喷洒20g处理液,(所述处理液含有1%葡萄糖,0.1%硫酸铵,0.1%七水硫酸镁,0.1%氯化钠,0.05%七水硫酸亚铁),在115℃下灭菌5min。
C.玳玳花发酵
将活化酵母种子液和根霉分别按照1:10和1:5的质量比接种到无菌的玳玳花原料中,搅拌,使原料与种子液充分混合均匀,放置于25℃的培养箱中发酵7天。
D.发酵后玳玳花挥发油提取
取发酵后的玳玳花20g,用滤纸包成小包放入索氏提取器中,提取瓶内加入石油醚100mL,然后固定在铁架台上,置于60℃恒温水浴锅中,同时冷凝水回流,提取至提取器中液体无色,取出提取液,加无水硫酸钠低温干燥24h过滤,旋转蒸发挥去滤液中的石油醚,收集挥发油。
实施例2
A.发酵菌种活化
①酵母菌活化:取酿酒酵母,接种于准备好的YPD斜面培养基上,放置于28℃的恒温培养箱中培养48h使菌种活化,再将活化的菌种接种于YPD液体培养基中,于28℃恒温培养48h,得到活化的种子液。YPD培养基:葡萄糖20g,酵母膏10g,蛋白胨20g,蒸馏水100mL(斜面培养基需加琼脂15g),以115℃灭菌20min。
②根霉活化:选取根霉菌种,接种于准备好的豆芽汁斜面培养基上,放置于25~30℃的恒温培养箱中培养48~72h使菌种活化,再将活化的菌种接种于YPD液体培养基中,于25~30℃恒温培养48~72h,得到活化的种子液;
B.玳玳花发酵原料预处理
选取200g玳玳花作为原料,用喉头喷雾器均匀喷洒40g水溶液,(含有1.5%葡萄糖,0.5%硫酸铵,0.28%七水硫酸镁,0.8%氯化钠,0.05%七水硫酸亚铁),在115℃下灭菌15min。
C.玳玳花发酵
将活化酵母种子液和根霉分别按照1:15和1:8的质量比接种到无菌的玳玳花原料中,搅拌,使原料与种子液充分混合均匀,放置于30的培养箱中发酵5天。
D.发酵后玳玳花挥发油提取
取发酵后的玳玳花20g,用滤纸包成小包放入索氏提取器中,提取瓶内加入石油醚100mL,然后固定在铁架台上,置于恒温水浴锅中(55℃),同时冷凝水回流,提取至提取器中液体无色,取出提取液,加无水硫酸钠低温干燥24h过滤,旋转蒸发挥去滤液中的石油醚,收集挥发油。
实施例3
A.发酵菌种活化
①酵母菌活化:取酿酒酵母,接种于准备好的YPD斜面培养基上,放置于30℃的恒温培养箱中培养48h使菌种活化,再将活化的菌种接种于YPD液体培养基中,于30℃恒温培养48h,得到活化的种子液。YPD培养基:葡萄糖20g,酵母膏10g,蛋白胨20g,蒸馏水100mL(斜面培养基需加琼脂15g),以115℃灭菌20min。
②根霉活化:选取根霉菌种,接种于准备好的豆芽汁斜面培养基上,放置于25~30℃的恒温培养箱中培养48~72h使菌种活化,再将活化的菌种接种于YPD液体培养基中,于25~30℃恒温培养48~72h,得到活化的种子液;
B.玳玳花发酵原料预处理
选取200g玳玳花作为原料,用喉头喷雾器均匀喷洒20~50g水溶液,(含有5%葡萄糖,1%硫酸铵,0.5%七水硫酸镁,1%氯化钠,0.12%七水硫酸亚铁),在115℃下灭菌15min。
C.玳玳花发酵
将活化酵母种子液和根霉分别按照1:20和1:10的质量比接种到无菌的玳玳花原料中,搅拌,使原料与种子液充分混合均匀,放置于32℃的培养箱中发酵7天。
D.发酵后玳玳花挥发油提取
取发酵后的玳玳花20g,用滤纸包成小包放入索氏提取器中,提取瓶内加入石油醚200mL,然后固定在铁架台上,置于恒温水浴锅中(60℃),同时冷凝水回流,提取至提取器中液体无色,取出提取液,加无水硫酸钠低温干燥24h过滤,旋转蒸发挥去滤液中的石油醚,收集挥发油。
实施例4
取在温度22℃±1℃和相对湿度(RH)60%±2%下平衡好的单料烟丝10g若干份,按其质量的0.01%、0.02%、0.05%分别称取玳玳花挥发油,用适量的95%的乙醇稀释后,用喉头喷管均匀的喷在烟丝上,加香的烟丝样品放入温度22℃±1℃和相对湿度(RH)60%±2%的恒温恒湿箱中平衡48h,用填烟器分别将加香的烟丝制成烟支,再在温度22℃±1℃和相对湿度(RH)60%±2%的恒温恒湿箱中平衡48h,请专业的卷烟感官评吸小组进行评吸。
评吸效果如表一所示。
表一 卷烟添加玳玳花烟用香料的评价效果
实施例5
玳玳花挥发油挥发性成分分析
①色谱条件:采用气相色谱-质谱联用技术(Gas chromatography-massspectrometry,GC/MS)对提取物进行分析;色谱柱为HP-5MS,规格为50cm×0.25mm×0.25μm;载气为氮气,进气量1.0mL/min;进样口温度260℃;升温速度50℃/min,5min升到250℃;进样量1μL,不分流;IE离子源,70eV,扫描范围50~650amu,传输线温度250℃;使用WILEY和MAINLIB谱库检索。
②挥发油前处理:取0.1g提取物,加入10ml无水乙醇,使其充分溶解,静置2h,取上清液,利用0.4um的微孔过滤器过滤,滤液放置于GC/MS自动进样器中进行分析。
分析结果:如表二所示是玳玳花挥发油挥发性成分分析结果,图一所示的是玳玳花挥发油挥发性成分分析GC/MS图谱。
表二 玳玳花挥发油挥发性成分分析结果
微生物在增殖过程中可产生庞大的高活性酶系,如多糖水解酶类、蛋白酶类、纤维素酶类、酯化酶类、氧化还原酶类及裂合酶类等,在酶促作用、化学作用及微生物体内复杂代谢的协同作用下,以玳玳花为原料发生分解、降解、氧化、还原、聚合、偶联、转化等作用,形成复杂的低分子化合物,其中包括各种香味化合物,如醇类、醛类、酮类、酸类、酯类、酚类、呋喃类、吡嗪类、吡啶类和萜烯类等,这些香气物质无疑也是烟草的香气组分。
本发明的香料对卷烟的香气和吸味品质改善作用明显,香气逼真,强度高,与香烟协调。本发明的方法工艺过程简单易行,设备简单,可直接用于卷烟加香加料,生产的产品可溶性好,便于推广应用。本发明所用的原料来源广泛,成本低,价格低廉,经济效益好。
Claims (5)
1.一种玳玳花烟用香料的制备方法,其特征在于,按照如下步骤进行:
A.发酵菌种活化
①酵母菌种活化:选取微生物菌种,接种于准备好的YPD斜面培养基上,放置于25~30℃的恒温培养箱中培养24~48h使菌种活化,再将活化的菌种接种于YPD液体培养基中,于25~30℃恒温培养24~48h,得到活化的种子液;
②根霉活化:选取根霉菌种,接种于准备好的豆芽汁斜面培养基上,放置于25~30℃的恒温培养箱中培养48~72h使菌种活化,再将活化的菌种接种于YPD液体培养基中,于25~30℃恒温培养48~72h,得到活化的种子液;
B.玳玳花发酵原料预处理
选取清洁干燥的玳玳花作为原料,均匀喷洒玳玳花质量0.1~0.25倍的处理液,在115℃下灭菌5~15min,即得玳玳花原料;所述处理液含有质量分数1~5%葡萄糖,0.1~1%硫酸铵,0.1~0.5%七水硫酸镁,0.1~1%氯化钠,0.05~0.12%七水硫酸亚铁,余量为水;
C.玳玳花发酵
将活化酵母种子液和根霉分别按照1:10~20和1:5~10的质量比接种到无菌的玳玳花原料中,搅拌,使原料与种子液充分混合均匀,放置于25~32℃的培养箱中发酵2~7天;
D.发酵后玳玳花挥发油提取
取发酵后的玳玳花,用滤纸包成小包放入索氏提取器中,提取瓶内加入石油醚,55~60℃回流提取,提取至提取器中液体至无色,取出提取液,加无水硫酸钠低温干燥24h过滤,挥去滤液中的石油醚,即为所述的玳玳花烟用香料。
2.根据权利要求1所述的一种玳玳花烟用香料的制备方法,其特征在于:优选的,步骤A中所述的酵母菌种为酿酒酵母菌,根霉菌种为白地根霉。
3.根据权利要求1所述的一种玳玳花烟用香料的制备方法,其特征在于:优选的,步骤A中所述的YPD培养基为:葡萄糖20g,酵母膏10g,蛋白胨20g,蒸馏水100mL;如果选用斜面培养基则需加琼脂15g;配制完成后以115℃灭菌20min。
4.根据权利要求1所述的一种玳玳花烟用香料的制备方法,其特征在于:优选的,步骤A中所述的豆芽汁斜面培养基为:10%豆芽汁20ml,蔗糖5g,纯净水80ml,121℃灭菌20min。
5.根据权利要求2所述的一种玳玳花烟用香料的制备方法,其特征在于:优选的,所述步骤D中石油醚体积与发酵后的玳玳花的质量比为5~10g/mL。
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