CN104894167A - Extraction method of Sparassis crispa fermentation product extract for cigarettes as well as application of extract - Google Patents
Extraction method of Sparassis crispa fermentation product extract for cigarettes as well as application of extract Download PDFInfo
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- CN104894167A CN104894167A CN201510340435.5A CN201510340435A CN104894167A CN 104894167 A CN104894167 A CN 104894167A CN 201510340435 A CN201510340435 A CN 201510340435A CN 104894167 A CN104894167 A CN 104894167A
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- 238000000605 extraction Methods 0.000 title claims abstract description 9
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- 125000004494 ethyl ester group Chemical group 0.000 description 1
- FMMOOAYVCKXGMF-MURFETPASA-N ethyl linoleate Chemical compound CCCCC\C=C/C\C=C/CCCCCCCC(=O)OCC FMMOOAYVCKXGMF-MURFETPASA-N 0.000 description 1
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- Manufacture Of Tobacco Products (AREA)
Abstract
The invention provides an extraction method of a Sparassis crispa fermentation product extract for cigarettes. The method comprises steps as follows: (1) firstly, Sparassis crispa fruiting bodies are inoculated, Sparassis crispa inoculum is obtained, then a fermentation medium is inoculated with the Sparassis crispa inoculum, submerged fermentation is performed, and a Sparassis crispa fermentation mixture is obtained; (2) reflux extraction is performed on the Sparassis crispa fermentation mixture obtained in Step (1) with ethyl alcohol with the mass concentration ranging from 90% to 95%, then filtration is performed, a filtrate is concentrated, and the Sparassis crispa fermentation product extract for the cigarettes is obtained. A submerged fermentation product of Sparassis crispa is extracted to be applied to essence and flavors for the cigarettes, the extract is natural, non-toxic and high in controllability and is not affected by factors such as seasons, weather, diseases and the like, the yield is stable, the production cycle is short, and the cost is low. Meanwhile, the prepared essence and flavors for the cigarettes are more natural and lasting in aroma, has better coordination performance, and can coordinate and cooperate with other essence and flavors for the cigarettes for aroma enhancement. The technology is simple, the cost is low, and the method has the very high practicability.
Description
Technical field
The present invention relates to the preparation field of essence spice for cigarette, specifically a kind of extracting method of Sparassis crispa fermented product cigarette extract and application thereof.
Background technology
Sparassis crispa is Basidiomycetes, and silk ball Cordycepps fungi, has another name called silk ball mushroom, and height above sea level growth scope is 2500-3700, is mainly distributed in the ground such as Heilungkiang, Jilin, Hebei, Yunnan, Shaanxi, Sichuan, Gansu, Tibet, Guangdong, Fujian.The every hectogram of Sparassis crispa is containing protein 15.58%, lipase 37 .95%, macromolecule polysaccharide 58%, N.F,USP MANNITOL 12%, carbohydrate 5%, the nutritive ingredient such as multivitamin and multiple amino acids is a kind of high tonic, there is strong essence, kidney tonifying, promote the production of body fluid, moistening lung, refreshing and brain nourishing, the function of antifatigue.It is rich in the polymer silk ball dextran of more than 60%, this special composition is the choice drug of Therapeutic cancer, except treat various cancer, tumour and except, also there is the immunologic function improving human body, function such as treatment cardiovascular and cerebrovascular diseases, diabetes, hypertension etc., ediblely can reduce cholesterol, the effect of reduce blood pressure, remove facial colour spot, pigment, freckle, macula lutea and skin-softening beauty for a long time.Research application at present for Sparassis crispa mainly concentrates on edible and medicinal aspect, also furthers investigate the extraction application of its tunning.
Essence and flavoring agent is usually used in the industry such as food, makeup, medicine, washing composition, and spices can be divided into natural perfume, natural equivalent spices and synthetic flavour 3 kinds.Because natural perfume is compared, other spices have fragrance nature, lasting, Harmony is good, single pure, and avoid heavy metal and organism solvent to advantages such as the harm of human body, for a long time, enjoy liking and paying close attention to of people always.In recent years, the demand of natural perfume sharply increases, and makes natural perfume be faced with animals and plants Flavour resources source more in short supply.Therefore, find the product perfume source that inexpensive, aboundresources, supply are stable, become the active demand of natural perfume development research.Sparassis crispa is crisp, fragrant and pleasant to taste, enjoys endless aftertastes, unique style, and like fennel taste, aromatic flavour is unique, has the potential characteristic preparing natural perfume.The wild resource of Sparassis crispa is on the increase with demand and is faced exhaustion gradually, and artificial culture because of output little and unstable, the application in fragrance industry is actually rare.Submerged fermentation (Submerged Fermentation, SmF) technology is a kind of biotechnology of quick obtaining medicine edible fungi raw materials, and it is with short production cycle, and cost is low, is convenient to operation management and is more suitable for large-scale commercial production.In recent years, the efficacy study to its activeconstituents is concentrated on mostly to the research of Sparassis crispa, and very few for the flavour ingredient research of Sparassis crispa.At present, not yet about the tunning extract of Sparassis crispa submerged fermentation being used for the report of cigarette additive.
Summary of the invention
The object of the invention is the extracting method and the application thereof that provide a kind of Sparassis crispa fermented product cigarette extract according to prior art, this cigarette extract to be used in cigarette not only fragrance naturally lasting, Harmony is good, and this extract stable yield, and controllability is strong.
The present invention is achieved through the following technical solutions: a kind of extracting method of Sparassis crispa fermented product cigarette extract, its step comprises: the inoculation of Sparassis crispa sporophore is first obtained Sparassis crispa inoculum by (1), then the volume ratio 5-15% of Sparassis crispa inoculum and fermention medium is pressed, Sparassis crispa inoculum is inoculated in fermention medium, at temperature 25-30 DEG C, air flow is 0.5-1.5dm
3under/min, rotating speed 120-170r/min condition, cultivate 3-7 days, carry out submerged fermentation and obtain Sparassis crispa fermenting mixture;
(2) be filter the Sparassis crispa fermenting mixture mass concentration in step (1) after the alcohol reflux of 90-95%, and obtain described Sparassis crispa fermented product cigarette extract by concentrated for filtrate.
Preferably, the amount of alcohol added in described step (2) is 0.5-2 times of Sparassis crispa fermenting mixture quality, and reflux temperature is 85-90 DEG C, refluxing extraction 1-3 hour.
Preferably, the middle Sparassis crispa sporophore inoculation of described step (1) obtains Sparassis crispa inoculum and refers to, by Sparassis crispa sporophore aseptically from stem position clip tissue fragment, be inoculated in separation agar plate, after within quiescent culture 5-7 days at temperature 28-30 DEG C, covering with separation agar plate to mycelia, aseptically picking is with the agar 10-20 block of mycelia, be seeded to seed culture medium, at temperature 26-28 DEG C, shaking speed is under 150r/min condition, cultivate 5-7 days, obtain described Sparassis crispa inoculum.
Preferred, the formula of described separation agar plate is: potato: 200-300g/L, sucrose: 15-25g/L, agar powder: 20-30g/L.
Preferred, the formula of described seed culture medium is: potato: 200-300g/L, sucrose: 15-25g/L.
Preferred, the formula of described fermention medium is: potato: 200-300g/L, sucrose 20-40g/L, potassium primary phosphate: 0.5-1.5g/L, Sodium Selenite: 1-1.5g/L.
Another object of the present invention is to provide the application of above-mentioned Sparassis crispa fermented product cigarette extract, it is characterized in that: added in recombination tobacco leaf according to the ratio of the 0.01-0.6% of recombination tobacco leaf weight by Sparassis crispa fermented product cigarette extract.
The present invention is used in the essence and flavoring agent of cigarette after being extracted by the submerged fermentation thing of Sparassis crispa, not only Nantural non-toxic, stable yield, and controllability is strong, not by the impact of the factors such as season, weather, disease, and also with short production cycle, with low cost.Meanwhile, the essence spice for cigarette fragrance of preparation is more natural, lasting, has better Harmony, can with other essence spice for cigarette cooperation flavourings.Present invention process is simple, and cost is low, has very strong practicality.
Embodiment
By the following specific examples further illustrate the invention:
Embodiment 1
(1) by Sparassis crispa sporophore aseptically from stem position clip fritter tissue fragment, be inoculated in separation agar plate, temperature 28 DEG C of quiescent culture 7 days, covering with after flat board until mycelia, aseptically with inoculating hook picking with 10 pieces, the agar of mycelia, being seeded to seed culture medium, temperature 28 DEG C, under shaking speed 150r/min condition, cultivate 5 days, obtain Sparassis crispa inoculum.10% Sparassis crispa inoculum is inoculated in fermention medium by volume, temperature 30 DEG C, air flow is 1.0dm
3under/min, rotating speed 150r/min condition, cultivate 7 days, carry out submerged fermentation and obtain Sparassis crispa fermenting mixture;
(2) 2: 1 ethanol that are 95% by Sparassis crispa tunning and mass concentration mix and add in water distilling apparatus by volume, distillation and reflux extraction 1h is carried out at 90 DEG C of temperature, pass through filter centrifugation, obtain extracting solution solution, the centrifugal concentrating that then reduces pressure obtains described amber transparent concrete Sparassis crispa fermented product cigarette extract 1.
Embodiment 2
(1) by Sparassis crispa sporophore aseptically from stem position clip fritter tissue fragment, be inoculated in separation agar plate, temperature 30 DEG C of quiescent culture 5 days, covering with after flat board until mycelia, aseptically with inoculating hook picking with 15 pieces, the agar of mycelia, being seeded to seed culture medium, temperature 26 DEG C, under shaking speed 150r/min condition, cultivate 6 days, obtain Sparassis crispa inoculum.15% Sparassis crispa inoculum is inoculated in fermention medium by volume, temperature 28 DEG C, air flow is 1.5dm
3under/min, rotating speed 120r/min condition, cultivate 5 days, carry out submerged fermentation and obtain Sparassis crispa fermenting mixture;
(2) 0.5: 1 ethanol that is 90% by Sparassis crispa tunning and mass concentration mixes and adds in water distilling apparatus by volume, distillation and reflux extraction 2h is carried out at 88 DEG C of temperature, pass through filter centrifugation, obtain extracting solution solution, the centrifugal concentrating that then reduces pressure obtains described amber transparent concrete Sparassis crispa fermented product cigarette extract 2.
Embodiment 3
(1) by Sparassis crispa sporophore aseptically from stem position clip fritter tissue fragment, be inoculated in separation agar plate, temperature 29 DEG C of quiescent culture 6 days, covering with after flat board until mycelia, aseptically with inoculating hook picking with 20 pieces, the agar of mycelia, being seeded to seed culture medium, temperature 27 DEG C, under shaking speed 150r/min condition, cultivate 7 days, obtain Sparassis crispa inoculum.5% Sparassis crispa inoculum is inoculated in fermention medium by volume, temperature 25 DEG C, air flow is 0.5dm
3under/min, rotating speed 170r/min condition, cultivate 3 days, carry out submerged fermentation and obtain Sparassis crispa fermenting mixture;
(2) 1: 1 ethanol that is 92% by Sparassis crispa tunning and mass concentration mixes and adds in water distilling apparatus by volume, distillation and reflux extraction 3h is carried out at 85 DEG C of temperature, pass through filter centrifugation, obtain extracting solution solution, the centrifugal concentrating that then reduces pressure obtains described amber transparent concrete Sparassis crispa fermented product cigarette extract 3.
In above-described embodiment 1-3, described separation agar plate formula is: potato: 200-300g/L, sucrose: 15-25g/L, and agar powder: 20-30g/L, after 121 DEG C of sterilizing 15min, aseptically pours sterilized petri dishes into.
Described seed culture based formulas is: potato: 200-300g/L, sucrose: 15-25g/L, filters potato cooking liquor obtain leach liquor, at 121 DEG C of autoclave sterilization 15min by double gauze.
The formula of described fermention medium is: potato 200-300g/L, adds distilled water water 1500ml, boiling water boiling 30 minutes, double gauze filters potato cooking liquor, sucrose 20-40g/L, potassium primary phosphate: 0.5-1.5g/L, Sodium Selenite 1-1.5g/L, at 121 DEG C of autoclave sterilization 15min.
Sparassis crispa fermented product cigarette extract in embodiment 1 is produced (6890N/5975C) Gas-phase acidity instrument with Agi lent company of the U.S. analyze, analysis condition is as follows, spectrogram as shown in Figure 1:
Chromatographic column: DB-WAX capillary column (30m × 250 μm × 0.25 μm); Carrier gas: He; Flow: 1.0mL/min; Splitting ratio: 3: 1; Sample size: 1 μ L; Heating schedule: injector temperature 250 DEG C, starting temperature 40 DEG C, stops 1min, rises to 250 DEG C, finally stop 15min with 10 DEG C/min.EI ionizing energy: 70eV; Ion source temperature: 250 DEG C; Mass spectrometer interface temperature: 280 DEG C; Quality of scanning scope: 40-500m/z.
Adopt NIST11 retrieval spectrum storehouse legal, areas of peak normalization method is quantitative.
Fig. 1 is the GC-MS figure of Sparassis crispa fermented product cigarette extract, goes out peak retention time and peak area result is as shown in table 1.Therefrom identify out 36 kinds of volatile compounds altogether, wherein aldoketones accounts for 5.866%, ester class (41.5%), the fragrant same clan (11.8%), olefines (14.2%), acids account for 1.636%, and alcohols accounts for 3.489, and other account for 2.503%.Wherein high the having of comparision contents: Laurate ethyl (1.762%), ethyl myristate (1.842%), 4-methyl, 4-pentenyl stupid (4.909), ethyl palmitate (0.958%), 13-methyl alkanoic acid ethyl ester (2.298%), 1,2,3,4-tetrahydrochysene 5,6,7,8-tetramethyl-naphthalene (1.498), Uniphat A60 (5.743), Stearic ethyl stearate (8.126), ethyl oleate (3.338), ethyl linoleate (10.789), 9,12,15-38 ethyl-carbonate (1.879), palmitinic acid (1.187).
Sparassis crispa fermented product cigarette extract is detected by GC-MS, and extract the material totally 36 kinds obtained, ester class has 15 kinds, olefines 13 kinds, the fragrance same clan has 8 kinds, and phenols has 3 kinds, and alcohols has 4 kinds, and aldoketones has 5 classes, fatty acid has 4 classes, and alkanes has 2 classes, and aldehydes has 1 class.
Some volatility mixing elements in Sparassis crispa fermented product cigarette extract, as aldehyde, ketone, acid and ester class, can to playing important soda acid mediation effect in cigarette local flavor, wherein aldehydes and ketone compounds are fragrance matters important in tobacco, generally there is happy fragrance, can give that tobacco nut, phenol are fragrant, the fragrance of aroma, aldehydes or a kind of very strong can local flavor effect equitant with other material, cigarette fragrance can be made more strong pure; Aromatics has amygdalate aromatic odour, and ketene has dense rose leaf fragrance, coordinates and close with tobacco flavor, thus strengthens the natural flavor of tobacco flavor; Add outward its ester class, alcohols and alkanes with fruital local flavor, thus make cigarette fragrance more natural, strong, various.Therefore, Sparassis crispa fermented product extract flavour ingredient is rich and varied, has fabulous cigarette fragrance-improving effect.
The GC-MS detected result of table 1 Sparassis crispa fermented product cigarette extract A.
Claims (7)
1. the extracting method of a Sparassis crispa fermented product cigarette extract, its step comprises: the inoculation of Sparassis crispa sporophore is first obtained Sparassis crispa inoculum by (1), then the volume ratio 5-15% of Sparassis crispa inoculum and fermention medium is pressed, Sparassis crispa inoculum is inoculated in fermention medium, at temperature 25-30 DEG C, air flow is 0.5-1.5dm
3under/min, rotating speed 120-170r/min condition, cultivate 3-7 days, carry out submerged fermentation and obtain Sparassis crispa fermenting mixture;
(2) be filter the Sparassis crispa fermenting mixture mass concentration in step (1) after the alcohol reflux of 90-95%, and obtain described Sparassis crispa fermented product cigarette extract by concentrated for filtrate.
2. the extracting method of Sparassis crispa fermented product cigarette extract according to claim 1, it is characterized in that: the amount of alcohol added in described step (2) is 0.5-2 times of Sparassis crispa fermenting mixture quality, reflux temperature is 85-90 DEG C, refluxing extraction 1-3 hour.
3. the extracting method of Sparassis crispa fermented product cigarette extract according to claim 1, it is characterized in that: the middle Sparassis crispa sporophore inoculation of described step (1) obtains Sparassis crispa inoculum and refers to, by Sparassis crispa sporophore aseptically from stem position clip tissue fragment, be inoculated in separation agar plate, after within quiescent culture 5-7 days at temperature 28-30 DEG C, covering with separation agar plate to mycelia, aseptically picking is with the agar 10-20 block of mycelia, be seeded to seed culture medium, at temperature 26-28 DEG C, shaking speed is under 150r/min condition, cultivate 5-7 days, obtain described Sparassis crispa inoculum.
4. the extracting method of Sparassis crispa fermented product cigarette extract according to claim 3, is characterized in that: the formula of described separation agar plate is: potato: 200-300g/L, sucrose: 15-25g/L, agar powder: 20-30g/L.
5. the extracting method of Sparassis crispa fermented product cigarette extract according to claim 3, is characterized in that: the formula of described seed culture medium is: potato: 200-300g/L, sucrose: 15-25g/L.
6. the extracting method of Sparassis crispa fermented product cigarette extract according to claim 1, it is characterized in that: the formula of described fermention medium is: potato: 200-300g/L, sucrose 20-40g/L, potassium primary phosphate: 0.5-1.5g/L, Sodium Selenite: 1-1.5g/L.
7. an application for Sparassis crispa fermented product cigarette extract according to any one of claim 1-6, is characterized in that: added in recombination tobacco leaf according to the ratio of the 0.01-0.6% of recombination tobacco leaf weight by Sparassis crispa fermented product cigarette extract.
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| CN112624947A (en) * | 2021-01-25 | 2021-04-09 | 福建万菇生物科技有限公司 | A method for extracting high content vitamin D from Sparassis crispa fruiting body2Method (2) |
| CN115141864A (en) * | 2022-09-06 | 2022-10-04 | 山东久茸生物科技有限公司 | Processing method of selenium-rich sparassis crispa polysaccharide |
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