CN107802025B - Preparation method of ginkgo leaf extract and application of ginkgo leaf extract in cigarettes - Google Patents
Preparation method of ginkgo leaf extract and application of ginkgo leaf extract in cigarettes Download PDFInfo
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- CN107802025B CN107802025B CN201711273705.0A CN201711273705A CN107802025B CN 107802025 B CN107802025 B CN 107802025B CN 201711273705 A CN201711273705 A CN 201711273705A CN 107802025 B CN107802025 B CN 107802025B
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- 235000019504 cigarettes Nutrition 0.000 title claims abstract description 51
- 239000009429 Ginkgo biloba extract Substances 0.000 title claims abstract description 37
- 238000002360 preparation method Methods 0.000 title claims abstract description 12
- 235000008100 Ginkgo biloba Nutrition 0.000 claims abstract description 68
- 235000011201 Ginkgo Nutrition 0.000 claims abstract description 67
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 55
- 238000001914 filtration Methods 0.000 claims abstract description 20
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 20
- 238000002156 mixing Methods 0.000 claims abstract description 18
- 238000000855 fermentation Methods 0.000 claims abstract description 15
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- 108010059892 Cellulase Proteins 0.000 claims description 10
- 108010047754 beta-Glucosidase Proteins 0.000 claims description 10
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- 229940106157 cellulase Drugs 0.000 claims description 10
- 230000000694 effects Effects 0.000 claims description 10
- 239000003795 chemical substances by application Substances 0.000 claims description 7
- 229940068052 ginkgo biloba extract Drugs 0.000 claims description 7
- 235000020686 ginkgo biloba extract Nutrition 0.000 claims description 7
- 238000000874 microwave-assisted extraction Methods 0.000 claims description 7
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- OVSQVDMCBVZWGM-LQSBFMDOSA-N 2-(3,4-dihydroxyphenyl)-5,7-dihydroxy-3-[(2r,3s,4r,5r,6s)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxychromen-4-one Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@@H]1OC1=C(C=2C=C(O)C(O)=CC=2)OC2=CC(O)=CC(O)=C2C1=O OVSQVDMCBVZWGM-LQSBFMDOSA-N 0.000 description 1
- 241000185686 Apocynum venetum Species 0.000 description 1
- JMGZEFIQIZZSBH-UHFFFAOYSA-N Bioquercetin Natural products CC1OC(OCC(O)C2OC(OC3=C(Oc4cc(O)cc(O)c4C3=O)c5ccc(O)c(O)c5)C(O)C2O)C(O)C(O)C1O JMGZEFIQIZZSBH-UHFFFAOYSA-N 0.000 description 1
- 241000218631 Coniferophyta Species 0.000 description 1
- UBSCDKPKWHYZNX-UHFFFAOYSA-N Demethoxycapillarisin Natural products C1=CC(O)=CC=C1OC1=CC(=O)C2=C(O)C=C(O)C=C2O1 UBSCDKPKWHYZNX-UHFFFAOYSA-N 0.000 description 1
- 229940123457 Free radical scavenger Drugs 0.000 description 1
- 241000218791 Ginkgoaceae Species 0.000 description 1
- FURUXTVZLHCCNA-UHFFFAOYSA-N Liquiritigenin Natural products C1=CC(O)=CC=C1C1OC2=CC(O)=CC=C2C(=O)C1 FURUXTVZLHCCNA-UHFFFAOYSA-N 0.000 description 1
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- 235000008777 kaempferol Nutrition 0.000 description 1
- MWDZOUNAPSSOEL-UHFFFAOYSA-N kaempferol Natural products OC1=C(C(=O)c2cc(O)cc(O)c2O1)c3ccc(O)cc3 MWDZOUNAPSSOEL-UHFFFAOYSA-N 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
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- FURUXTVZLHCCNA-AWEZNQCLSA-N liquiritigenin Chemical compound C1=CC(O)=CC=C1[C@H]1OC2=CC(O)=CC=C2C(=O)C1 FURUXTVZLHCCNA-AWEZNQCLSA-N 0.000 description 1
- KBPHJBAIARWVSC-RGZFRNHPSA-N lutein Chemical compound C([C@H](O)CC=1C)C(C)(C)C=1\C=C\C(\C)=C\C=C\C(\C)=C\C=C\C=C(/C)\C=C\C=C(/C)\C=C\[C@H]1C(C)=C[C@H](O)CC1(C)C KBPHJBAIARWVSC-RGZFRNHPSA-N 0.000 description 1
- 229960005375 lutein Drugs 0.000 description 1
- ORAKUVXRZWMARG-WZLJTJAWSA-N lutein Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CCCC1(C)C)C=CC=C(/C)C=CC2C(=CC(O)CC2(C)C)C ORAKUVXRZWMARG-WZLJTJAWSA-N 0.000 description 1
- 235000012680 lutein Nutrition 0.000 description 1
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- 239000003094 microcapsule Substances 0.000 description 1
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- 230000004048 modification Effects 0.000 description 1
- UXOUKMQIEVGVLY-UHFFFAOYSA-N morin Natural products OC1=CC(O)=CC(C2=C(C(=O)C3=C(O)C=C(O)C=C3O2)O)=C1 UXOUKMQIEVGVLY-UHFFFAOYSA-N 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
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- 150000007524 organic acids Chemical class 0.000 description 1
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- 150000008442 polyphenolic compounds Chemical class 0.000 description 1
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- 239000000047 product Substances 0.000 description 1
- FDRQPMVGJOQVTL-UHFFFAOYSA-N quercetin rutinoside Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC=2C(C3=C(O)C=C(O)C=C3OC=2C=2C=C(O)C(O)=CC=2)=O)O1 FDRQPMVGJOQVTL-UHFFFAOYSA-N 0.000 description 1
- 239000002516 radical scavenger Substances 0.000 description 1
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- 238000005096 rolling process Methods 0.000 description 1
- 235000005493 rutin Nutrition 0.000 description 1
- IKGXIBQEEMLURG-BKUODXTLSA-N rutin Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@@H]1OC[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](OC=2C(C3=C(O)C=C(O)C=C3OC=2C=2C=C(O)C(O)=CC=2)=O)O1 IKGXIBQEEMLURG-BKUODXTLSA-N 0.000 description 1
- ALABRVAAKCSLSC-UHFFFAOYSA-N rutin Natural products CC1OC(OCC2OC(O)C(O)C(O)C2O)C(O)C(O)C1OC3=C(Oc4cc(O)cc(O)c4C3=O)c5ccc(O)c(O)c5 ALABRVAAKCSLSC-UHFFFAOYSA-N 0.000 description 1
- 229960004555 rutoside Drugs 0.000 description 1
- 230000002000 scavenging effect Effects 0.000 description 1
- 238000000638 solvent extraction Methods 0.000 description 1
- GSZUGBAEBARHAW-UHFFFAOYSA-N sophoraflavone B Natural products OC1C(O)C(O)C(CO)OC1OC1=CC=C(C=2OC3=CC(O)=CC=C3C(=O)C=2)C=C1 GSZUGBAEBARHAW-UHFFFAOYSA-N 0.000 description 1
- 239000004575 stone Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 235000019505 tobacco product Nutrition 0.000 description 1
- KBPHJBAIARWVSC-XQIHNALSSA-N trans-lutein Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CC(O)CC1(C)C)C=CC=C(/C)C=CC2C(=CC(O)CC2(C)C)C KBPHJBAIARWVSC-XQIHNALSSA-N 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
- FJHBOVDFOQMZRV-XQIHNALSSA-N xanthophyll Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CC(O)CC1(C)C)C=CC=C(/C)C=CC2C=C(C)C(O)CC2(C)C FJHBOVDFOQMZRV-XQIHNALSSA-N 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A24—TOBACCO; CIGARS; CIGARETTES; SIMULATED SMOKING DEVICES; SMOKERS' REQUISITES
- A24B—MANUFACTURE OR PREPARATION OF TOBACCO FOR SMOKING OR CHEWING; TOBACCO; SNUFF
- A24B3/00—Preparing tobacco in the factory
- A24B3/12—Steaming, curing, or flavouring tobacco
-
- A—HUMAN NECESSITIES
- A24—TOBACCO; CIGARS; CIGARETTES; SIMULATED SMOKING DEVICES; SMOKERS' REQUISITES
- A24B—MANUFACTURE OR PREPARATION OF TOBACCO FOR SMOKING OR CHEWING; TOBACCO; SNUFF
- A24B15/00—Chemical features or treatment of tobacco; Tobacco substitutes, e.g. in liquid form
- A24B15/18—Treatment of tobacco products or tobacco substitutes
- A24B15/24—Treatment of tobacco products or tobacco substitutes by extraction; Tobacco extracts
- A24B15/241—Extraction of specific substances
-
- A—HUMAN NECESSITIES
- A24—TOBACCO; CIGARS; CIGARETTES; SIMULATED SMOKING DEVICES; SMOKERS' REQUISITES
- A24B—MANUFACTURE OR PREPARATION OF TOBACCO FOR SMOKING OR CHEWING; TOBACCO; SNUFF
- A24B15/00—Chemical features or treatment of tobacco; Tobacco substitutes, e.g. in liquid form
- A24B15/18—Treatment of tobacco products or tobacco substitutes
- A24B15/24—Treatment of tobacco products or tobacco substitutes by extraction; Tobacco extracts
- A24B15/26—Use of organic solvents for extraction
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B9/00—Essential oils; Perfumes
- C11B9/0003—Compounds of unspecified constitution defined by the chemical reaction for their preparation
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
- C12R2001/125—Bacillus subtilis ; Hay bacillus; Grass bacillus
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- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Wood Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- General Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
- General Health & Medical Sciences (AREA)
- Mycology (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
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- Medicines Containing Plant Substances (AREA)
Abstract
The invention provides a preparation method of a ginkgo leaf extract and application thereof in cigarettes, comprising the following steps: (1) cleaning fresh folium Ginkgo, draining, slicing, extracting with ethanol solution under microwave, filtering, and collecting filtrate to obtain folium Ginkgo extractive solution A. (2) Collecting the ginkgo leaf residues in the step (1), adding water and biological agents, hydrolyzing and fermenting, adding ethanol into hydrolysis fermentation liquor, carrying out reflux extraction at constant temperature, and filtering to obtain a ginkgo leaf extracting solution B. (3) And mixing the ginkgo leaf extract A and the ginkgo leaf extract B, and concentrating under reduced pressure to obtain the ginkgo leaf extract. The ginkgo leaf extract for cigarettes prepared by the invention has the characteristic of the unique herb aroma of ginkgo leaves, is applied to cigarettes as an additive, is coordinated with the cigarette aroma, enriches the cigarette aroma, effectively solves the negative problems of high irritation and more oral residues easily caused by the conventional extraction process, and has the function of removing the free radical content in the mainstream smoke of the cigarettes.
Description
Technical Field
The invention relates to the field of cigarette flavors and fragrances, in particular to a preparation method of a ginkgo leaf extract and application thereof in cigarettes.
Background
Ginkgo (also known as Gongsun tree and gingko biloba) is a plant of the family Ginkgoaceae of the gymnosperm, and is one of the oldest tree species on earth today. Ginkgo biloba is a name of pandas in the plant world, is an activated stone for researching ancient geological deep change of the earth, and is listed as a second-level rare protection tree species by the nation. The gingko resource in China accounts for about 75% of the total amount of the world and is the first place in the world. Ginkgo in most areas of China is artificially cultivated and mainly distributed in provinces and cities of Hubei, Zhejiang, Anhui, Fujian, Henan, Sichuan and the like. Thousands of old ginkgo trees grow in the deep mountains of the city of the northwest of Hu, which is one of four dense ancient ginkgo communities in the world.
When the cigarette is burnt, high-concentration free radicals are generated, and the free radicals can directly or indirectly attack and damage substances such as cells, enzymes and the like and are combined with DNA to transform the cells to generate mutation. Therefore, the research on free radicals in smoke and the development of the efficient and nontoxic free radical scavenger have very important significance for reducing the smoking harm of cigarettes and relieving the contradiction between smoking and health. At present, researches on reducing free radicals of smoke by adopting plant additives are few, and apocynum venetum, tea polyphenol, liquiritigenin, lutein and the like which are reported as cigarette additives have the effect of reducing free radicals, and the reduction range is about 7-20%. The adding process of the plant additives is mostly to be made into microcapsules to be added into the filter tip, and the process is complex. This is mainly because, firstly, most of the plant additives are fat-soluble and have difficulty in the process of directly spraying the plant additives into the cut tobacco; and secondly, the plant additives are added into tobacco shreds, so that the smoking taste of cigarettes is changed, and even very bad influence is brought. There are many natural plants capable of reducing free radicals, but the natural plants capable of reducing free radicals of cigarettes simultaneously can bring good positive effect to the taste of the cigarettes. Although a lot of natural plants for reducing free radicals of cigarettes are reported at present, almost none of the natural plants are really used for industrial production and bring direct and remarkable economic benefits, and the key reason is that the negative effect of the plant additives on the smoking taste of the cigarettes cannot be solved.
The ginkgo leaf is used as a Chinese herbal medicine and has various biological activities and medicinal values, such as the effects of resisting oxidation, removing free radicals of a human body, improving blood flow, increasing cerebral blood supply and the like. The main medicinal components of ginkgo leaf are flavone and lactone compounds (the content is greater than 24.0%), the flavone compounds currently found comprise rutin, astragalus root, isoquercitin, kaempferol and the like, and the lactone compounds comprise bilobalide and bilobalide, can endow unique herb smell, and have the prospect of being used as essence and spice for cigarettes.
In recent years, the function of natural plant spices in improving the comfort of cigarettes and highlighting the style characteristics of cigarettes is widely accepted by tobacco technologists and consumers. At present, ginkgo leaves are mostly used in the field of medicine in the research results reported in literature. The inventor finds that the ginkgo leaf extract obtained by a simple solvent extraction process has the disadvantages that due to high extraction temperature, part of volatile components of ketones and esters are easy to be oxidized and decomposed, so that the aroma is lost, the characteristic aroma of the product is not obvious, the natural feeling is not ideal, and the ginkgo leaf extract added into cigarettes easily brings negative effects of high irritation, more oral residues and the like, so that the ginkgo leaf extract cannot meet the use requirements of being used as an additive of tobacco products.
Disclosure of Invention
The invention aims to provide a preparation method of a ginkgo leaf extract and application of the ginkgo leaf extract in cigarettes according to the defects of the prior art, the extract prepared by the invention has the characteristic of the unique herb aroma of ginkgo leaves, the aroma is natural and harmonious, the natural sense of aroma is strong, the extract is applied to cigarettes as an additive, the personalized aroma is prominent, the extract is harmonious with the tobacco aroma, the cigarette aroma is enriched, the aroma quality is improved, the aftertaste is comfortable, the oral cavity is clean, the smoke is soft and fine, the negative problems of large irritation and more oral cavity residues easily caused by the conventional extraction process are effectively solved, and meanwhile, the content of free radicals in the mainstream smoke of the cigarettes can be eliminated.
In order to achieve the purpose, the preparation method of the ginkgo leaf extract and the application of the ginkgo leaf extract in cigarettes provided by the invention comprise the following steps:
(1) cleaning fresh ginkgo leaves, draining, slicing for later use, mixing the flaky ginkgo leaves with 30-60% ethanol solution according to the mass ratio of 1:5-10, putting the mixture into microwave extraction equipment for extraction, setting the microwave power at 350-700W, setting the system temperature to be 50-65 ℃ from room temperature within 10-30 min, processing for 0.5-1 h, filtering and collecting filtrate to obtain ginkgo leaf extracting solution A.
(2) Collecting the ginkgo leaf residues in the step (1), adding 2-5 times of water by mass, adding 0.1-0.5% of biological agent by mass, uniformly stirring, and carrying out incubation, hydrolysis and fermentation for 6-18h under the conditions of pH4-6 and temperature of 30-50 ℃ to obtain hydrolysis fermentation liquor. Adding food grade 95% ethanol (ethanol concentration of 30-60% in the system) into the hydrolysis fermentation broth, reflux-extracting for 1-3 times in constant temperature water bath at 60-80 deg.C, filtering, and mixing filtrates to obtain folium Ginkgo extract B.
(3) Mixing folium Ginkgo extractive solution A and folium Ginkgo extractive solution B, and concentrating under reduced pressure to 1/4. Adding 5-8 times of water into the system, stirring uniformly, standing for 6-12h, filtering supernatant, concentrating under reduced pressure to 1/5-1/3 of the original volume, thus obtaining the ginkgo leaf extract.
The fresh folium Ginkgo is derived from the valley of Ginkgo of Luzhou Luoyang.
The biological agent in the step (2) is one or more of beta-glucosidase, cellulase and bacillus subtilis live bacteria agent.
When the biological agent in the step (2) is compounded by two or more than two, the compounding proportion of the beta-glucosidase, the cellulase and the bacillus subtilis live bacteria agent is 1-2: 1-2: 0.2-1.
The enzyme activity of the beta-glucosidase and the cellulase is 5-20 ten thousand u/g, and the viable count of the bacillus subtilis contained in the viable bacillus subtilis preparation is not less than 1.0 x 108CFU/g。
The invention provides another technical scheme: the application of the ginkgo leaf extract prepared by the method is characterized in that: the ginkgo leaf extract is added into cigarettes as a natural cigarette additive in a proportion of 0.1-0.3% of the mass of tobacco shreds.
The specific steps of adding the ginkgo leaf extract into the cigarette tobacco shreds are as follows: dissolving folium Ginkgo extract in 70% ethanol, spraying the ethanol solvent containing folium Ginkgo extract on tobacco shreds during cigarette production, and evaporating the solvent.
The invention has the advantages that:
(1) the commercial ginkgo leaves are generally dried traditional Chinese medicinal materials, and the ginkgo leaves usually have the volatilization loss of aroma components in the drying and processing processes.
(2) The microwave extraction process has the technical advantages of low-temperature extraction and high extraction efficiency, reduces the oxidative decomposition of volatile aroma components such as ketones and esters, and effectively avoids aroma loss.
(3) The method utilizes the biological agent to carry out hydrolytic fermentation on the ginkgo leaf residues, degrades macromolecular substances into utilizable micromolecular substances, generates organic acids such as lactic acid and the like, improves the comprehensive utilization rate of raw materials, and avoids resource waste.
(4) The extract prepared by the invention has the characteristic of the unique herb aroma of ginkgo leaves, the aroma is natural and harmonious, the natural sense of the aroma is strong, the extract is used as an additive to be applied to cigarettes, the personalized aroma is prominent, the extract is harmonious with the tobacco aroma, the aroma quantity of the cigarettes is enriched, the aroma quality is improved, the aftertaste is comfortable, the oral cavity is clean, the smoke is soft and delicate, the negative problems of large irritation and more oral cavity residues easily caused by a conventional extraction process are effectively solved, and meanwhile, the extract has the function of removing the content of free radicals in the mainstream smoke of the cigarettes.
(5) The method has the characteristics of simple operation, easy control, low raw material cost and the like, is suitable for actual production conversion, and is suitable for popularization and application as a method for preparing the tobacco flavor.
Detailed Description
The present invention will be described in further detail with reference to specific examples.
Example one
(1) Cleaning fresh ginkgo leaves, draining, slicing for later use, mixing the flaky ginkgo leaves with 60% ethanol solution according to the proportion of 1:10, putting the mixture into microwave extraction equipment for extraction, setting the microwave power to be 550W, setting the system temperature to be 50-65 ℃ from room temperature within 10-30 min, processing for 0.5h, filtering, and collecting filtrate to obtain ginkgo leaf extracting solution A.
(2) Collecting the ginkgo leaf residues in the step (1), adding 5 times of water by mass, adding 0.5% of biological agent by mass (the compounding ratio of beta-glucosidase, cellulase and bacillus subtilis live bacteria agent is 1: 1: 0.5), uniformly stirring, and then incubating, hydrolyzing and fermenting for 6 hours under the conditions of pH4 and temperature of 30 ℃ to obtain hydrolysis fermentation liquor. Adding food grade 95% ethanol (alcohol concentration is 50% in the system) into the hydrolysis fermentation liquid, reflux-extracting for 3 times in a constant temperature water bath at 60 deg.C, filtering, and mixing filtrates to obtain folium Ginkgo extract B.
(3) Mixing folium Ginkgo extractive solution A and folium Ginkgo extractive solution B, and concentrating under reduced pressure to 1/4. Adding 8 times of water into the system, stirring uniformly, standing for 10h, filtering supernatant, and concentrating under reduced pressure to 1/3 of the original volume to obtain the ginkgo leaf extract.
Example two
(1) Cleaning fresh ginkgo leaves, draining, slicing for later use, mixing the flaky ginkgo leaves with 30% ethanol solution according to the proportion of 1:8, putting the mixture into microwave extraction equipment for extraction, setting the microwave power at 350W, setting the system temperature to be 50-65 ℃ from room temperature within 10-30 min, treating for 1h, filtering and collecting filtrate to obtain ginkgo leaf extracting solution A.
(2) Collecting the ginkgo leaf residues in the step (1), adding 4 times of water by mass, adding 0.1% of biological agent by mass (the compounding ratio of beta-glucosidase, cellulase and bacillus subtilis live bacteria agent is 1: 1: 1), uniformly stirring, and then incubating and hydrolyzing for 12h under the conditions of pH6 and temperature of 50 ℃ to obtain hydrolysis fermentation liquor. Adding food-grade 95% ethanol (ethanol concentration of 40% in the system) into the hydrolysis fermentation broth, reflux-extracting for 2 times in a constant-temperature water bath at 65 deg.C, filtering, and mixing filtrates to obtain folium Ginkgo extract B.
(3) Mixing folium Ginkgo extractive solution A and folium Ginkgo extractive solution B, and concentrating under reduced pressure to 1/4. Adding 5 times of water into the system, stirring uniformly, standing for 8h, filtering supernatant, and concentrating under reduced pressure to 1/4 of the original volume to obtain the ginkgo leaf extract.
EXAMPLE III
(1) Cleaning fresh ginkgo leaves, draining, slicing for later use, mixing the flaky ginkgo leaves with 45% ethanol solution according to the proportion of 1:6, putting the mixture into microwave extraction equipment for extraction, setting the microwave power at 450W, setting the system temperature to be 50-65 ℃ from room temperature within 10-30 min, processing for 0.7h, filtering, and collecting filtrate to obtain ginkgo leaf extracting solution A.
(2) Collecting the ginkgo leaf residues in the step (1), adding 3 times of water by mass, adding 0.3% of biological agent by mass (the compounding ratio of beta-glucosidase, cellulase and bacillus subtilis live bacteria agent is 1: 2: 0.5), uniformly stirring, and then incubating, hydrolyzing and fermenting for 9 hours at the temperature of 35 ℃ and the pH of 5.5 to obtain hydrolysis fermentation liquor. Adding food-grade 95% ethanol (alcohol concentration of 35% in the system) into the hydrolysis fermentation broth, reflux-extracting for 3 times in 70 deg.C constant temperature water bath, filtering, and mixing filtrates to obtain folium Ginkgo extract B.
(3) Mixing folium Ginkgo extractive solution A and folium Ginkgo extractive solution B, and concentrating under reduced pressure to 1/4. Adding 7 times of water into the system, stirring uniformly, standing for 7h, taking supernatant, filtering, and concentrating under reduced pressure to 1/3 of the original volume, namely the ginkgo leaf extract.
Example four
(1) Cleaning fresh ginkgo leaves, draining, slicing for later use, mixing the flaky ginkgo leaves with 55% ethanol solution according to the proportion of 1:8, putting the mixture into microwave extraction equipment for extraction, setting the microwave power at 650W, setting the system temperature to be 50-65 ℃ from room temperature within 10-30 min, processing for 0.8h, filtering, and collecting filtrate to obtain ginkgo leaf extracting solution A.
(2) Collecting the ginkgo leaf residues in the step (1), adding 4 times of water by mass, adding 0.4% of biological agent by mass (the compounding ratio of beta-glucosidase, cellulase and bacillus subtilis live bacteria agent is 1: 1: 0.4), uniformly stirring, and then incubating, hydrolyzing and fermenting for 16h under the conditions of pH4.5 and 40 ℃ to obtain hydrolysis fermentation liquor. Adding food grade 95% ethanol (alcohol concentration is 50% in the system) into the hydrolysis fermentation liquid, reflux-extracting for 2 times in 75 deg.C constant temperature water bath, filtering, and mixing filtrates to obtain folium Ginkgo extract B.
(3) Mixing folium Ginkgo extractive solution A and folium Ginkgo extractive solution B, and concentrating under reduced pressure to 1/4. Adding 8 times of water into the system, stirring uniformly, standing for 10h, filtering supernatant, and concentrating under reduced pressure to 1/5 of the original volume to obtain the ginkgo leaf extract.
Application test
During the process of rolling tobacco shreds in cigarette production, the ginkgo leaf extract is added into the tobacco shreds, and the ginkgo leaf extract accounts for 0.2 percent of the weight of the tobacco shreds. The specific method comprises dissolving folium Ginkgo extract in ethanol, spraying the ethanol solution on tobacco shreds uniformly, evaporating ethanol, and making tobacco shreds into cigarette sample by high-speed cigarette making machine. The tobacco was evaluated against a blank cigarette without the extract added thereto with reference to national bureau smoking standards. The smoking panel consisted of 13 sensory evaluation experts of cigarettes, and was subjected to sensory evaluation in a dark evaluation mode and scored, and the sensory evaluation results are shown in table 1.
TABLE 1 evaluation results of sensory Effect of cigarettes
The effect of eliminating free radical content in mainstream smoke of cigarettes after the ginkgo biloba extract is added is analyzed and detected by adopting a smoke collector and an electron spin resonance spectroscopy, and the result is shown in table 2.
TABLE 2 scavenging effect of Ginkgo biloba extract on free radical content of cigarette mainstream smoke
Note: increasing the content of free radicals; -, the content of free radicals is reduced
After the ginkgo leaf extract is added, the sensory effect evaluation score of the cigarette is obviously improved, the application effect is more obvious, the personalized aroma is prominent, the aroma is rich, full, pure and comfortable, the aroma is lasting, the aftertaste is clean and comfortable, the smoke is soft and delicate, and the flavoring effect is stable; meanwhile, the cigarette filter has the effect of removing the content of free radicals in the mainstream smoke of the cigarette in the burning and smoking process of the cigarette.
The above description is only a detailed description of specific embodiments of the present invention, and should not be taken as limiting the invention, and any modifications, equivalents, improvements, etc. made on the design concept of the present invention should be included in the protection scope of the present invention.
Claims (6)
1. A preparation method of a ginkgo leaf extract is characterized by comprising the following steps:
(1) cleaning fresh ginkgo leaves, draining, slicing for later use, mixing the flaky ginkgo leaves with 30-60% ethanol solution according to a mass ratio of 1:5-10, putting the mixture into microwave extraction equipment for extraction, setting the microwave power to be 350-700W, setting the system temperature to be 50-65 ℃ from room temperature within 10-30 min, processing for 0.5-1 h, filtering and collecting filtrate to obtain ginkgo leaf extracting solution A;
(2) collecting the ginkgo leaf residues in the step (1), adding 2-5 times of water by mass, adding a biological agent according to the mass ratio of 0.1-0.5% of the ginkgo leaf residues, uniformly stirring, incubating, hydrolyzing and fermenting for 6-18h under the conditions of pH4-6 and temperature of 30-50 ℃ to obtain hydrolysis fermentation liquor, adding food-grade 95% ethanol into the hydrolysis fermentation liquor, carrying out reflux extraction for 1-3 times in a constant-temperature water bath at 60-80 ℃, filtering, and combining filtrates to obtain a ginkgo leaf extracting solution B;
(3) mixing folium Ginkgo extract A and folium Ginkgo extract B, concentrating under reduced pressure to 1/4 of original volume, adding 5-8 times of water, stirring, standing for 6-12 hr, filtering supernatant, and concentrating under reduced pressure to 1/5-1/3 of original volume to obtain folium Ginkgo extract.
2. The method of claim 1, wherein the step of preparing the ginkgo biloba extract comprises: the biological agent in the step (2) is a compound preparation consisting of any one or two of beta-glucosidase and cellulase and a bacillus subtilis live bacterial agent.
3. The method of claim 2, wherein the step of preparing the ginkgo biloba extract comprises: the compounding ratio of the beta-glucosidase, the cellulase and the live bacillus subtilis preparation in the biological preparation in the step (2) is 1-2: 1-2: 0.2-1.
4. The method of claim 2, wherein the step of preparing the ginkgo biloba extract comprises: the enzyme activities of the beta-glucosidase and the cellulase in the step (2) are both 5-20 ten thousand u/g, and the viable count of the bacillus subtilis contained in the viable bacillus subtilis preparation is not less than 1.0 x 108CFU/g。
5. The method of claim 1, wherein the step of preparing the ginkgo biloba extract comprises: and (3) adding food-grade 95% ethanol in the step (2) to ensure that the mass concentration of the ethanol in the solution extracted by the constant-temperature water bath is 30-60%.
6. Use of the ginkgo biloba extract prepared according to any one of claims 1 to 5 in cigarettes, characterized in that: the ginkgo leaf extract is added into cigarettes as a natural cigarette additive in a proportion of 0.1-0.3 percent of the mass of tobacco shreds, and comprises the following steps: dissolving folium Ginkgo extract in ethanol, spraying the ethanol solution on tobacco shreds, evaporating ethanol, and making tobacco shreds into cigarette sample by high speed cigarette making machine.
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