CN101790938B - Method for improving content and extraction rate of flavonoids in fruit bodies of Stropharia rugoso-annulata - Google Patents
Method for improving content and extraction rate of flavonoids in fruit bodies of Stropharia rugoso-annulata Download PDFInfo
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Abstract
The invention relates to a method for improving the content and extraction rate of flavonoids in fruit bodies of Stropharia rugoso-annulata and belongs to the field of biotechnology. In the method, waste resources such as straw, husk and wooden meal are used as a culture medium of Stropharia rugoso-annulata to cultivate the Stropharia rugoso-annulata and conditions influencing the extraction of the flavonoids of the Stropharia rugoso-annulata are further optimized so as to improve the content and the extraction rate of the flavonoid compounds of the Stropharia rugoso-annulata. The formula of the Stropharia rugoso-annulata culture medium comprises 100 percent of husk or 50 percent of husk and 50 percent of straw, and the Stropharia rugoso-annulata is produced and grown in a mode of outdoor raw meal idyllized whole ridge cultivation to produce the fruit bodies of the Stropharia rugoso-annulata. The conditions for optimizing the extraction rate of the extraction rate of the flavonoids in the fruit bodies of the Stropharia rugoso-annulata comprise that: the material to solvent (W/V) ratio is 1:40-70; the extraction solvent is 80 to 90 (V) percent food grade industrial alcohol; the extraction time is 2 to 3 hours; and the extraction temperature is 70 to 90 DEG C. The method has the advantages of recycling waste, environmental protection, simply and conveniently operated extraction method, short production period and great economic benefits.
Description
Technical field
The present invention relates to a kind of method that improves flavonoid content in the Stropharia rugoso-annulata fruit body and extraction efficiency thereof, belong to biological technical field.
Background technology
Stropharia rugoso-annulata (
Stropharia rugoso-annulataFarlow) be under the jurisdiction of Basidiomycota (
Basidomycota), Hymenomycetes (
Hymenomycletes), Agaricales (
Agaricales), Stropharia rugoso-annulata section (
Strophariaceae), Stropharia (
Stropharia).Another name Stropharia rugoso annulata, wrinkle ring cowling mushroom, the red Stropharia rugoso-annulata of wine, Pei Shi Stropharia rugoso-annulata, the false black umbrella of Pei Shi.Being one of tame famous edible mushroom of European and American countries, also is that one of edible mushroom of cultivating is recommended by FAO (Food and Agriculture Organization of the United Nation) (FAO) to developing country.The Stropharia rugoso-annulata fine and tender taste, handle is crisp, lovely luster, delicious flavour, delicate fragrance is good to eat, and dried mushroom giving off a strong fragrance contains nutrient components such as abundant proteins,vitamins,minerals and polysaccharide, enjoys the good reputation of " mountain forest treasure ".Stropharia rugoso-annulata is not only the edible fungus that contains high nutritive value, still a kind of extremely precious medicinal fungi, and it has anti-oxidant, antitumor, prevention coronary heart disease, help digest, improve body immunity, alleviate medicinal efficacies such as mental fatigue.
In recent years, the anti-oxidant research of edible mushroom and secondary metabolite thereof has become the focus of present research and development, studies show that Stropharia rugoso-annulata is not only nutritious, and contains multiple anti-oxidation active substance.In addition, Stropharia rugoso-annulata has extremely strong degeneration-resistant anti-polygamy, is easy to artificial cultivation, the Stropharia rugoso-annulata aboundresources of China.Yet according to reported in literature, the flavonoid component content of Stropharia rugoso-annulata only is about 1 ‰ of its dry matter composition, has not yet to see the technical research bibliographical information that improves flavonoid content in the Stropharia rugoso-annulata fruit body.
The extracting method of flavonoids mainly contains three kinds at present: circumfluence method, microwave method and supercritical ultrasonics technology.Compare with supercritical ultrasonics technology with microwave method, circumfluence method has advantages such as equipment is simple, easy to operate, safety and environmental protection, though there is defective more consuming time in this technology, also provides and can carry out the potentiality that condition is preferably optimized.
Summary of the invention
The object of the present invention is to provide a kind of technology that improves flavonoid content in the Stropharia rugoso-annulata fruit body and extraction efficiency thereof.
In order to realize this purpose, the present invention finds that by test careless saprophytic edible mushroom Stropharia rugoso-annulata has degradation of rice straw, polyphenols and be converted into the biological utilisation potentiality of Flavonoid substances in the husk, the human Polyphenols lignin that all can't fully degrade with animal in can the Degradation and Transformation plant corpus, polysaccharose fiber and hemicellulose, become function nutrition active component relevant in the fruit body with oxidation resistance, therefore Stropharia rugoso-annulata contains the flavonoids of high level, the saponin class, antioxidation biology such as phenols and vitamin C active component is good natural anti-oxidation material source.
Technical scheme of the present invention is as follows:
㈠ is a foundation with the Nutrition and Metabolism characteristics of Stropharia rugoso-annulata, improves the prescription of stropharia rugoso-annulata plantation matrix, in the hope of reaching the purpose that improves flavonoid content in the Stropharia rugoso-annulata fruit body.The prescription of described stropharia rugoso-annulata plantation matrix is 100% husk or 50% husk+50% straw; Adopt the whole bedding cultivation mode of outdoor raw material garden farming to produce plantation, obtain the Stropharia rugoso-annulata fruit body, make that flavonoid content improves 10%～35% in the Stropharia rugoso-annulata fruit body.
㈡ improves flavonoids extraction yield in the Stropharia rugoso-annulata fruit body with anti-oxidation function by optimizing extraction conditions such as extracting solvent, material ratio, solvent strength and extraction time, extraction temperature.
1. with water, acetone, ethyl acetate, four kinds of solvents of ethanol as the extraction solvent that extracts flavonoids in the Stropharia rugoso-annulata fruit body, found that flavonoids efficient best solvent is an ethanol in the extraction Stropharia rugoso-annulata fruit body, therefore, flavonoids extracts with the food-grade industrial alcohol as extracting solvent in the Stropharia rugoso-annulata fruit body;
2. extraction conditions such as material ratio, solvent strength and extraction time, extraction temperature influences the extraction yield of flavonoids in the Stropharia rugoso-annulata fruit body.The optimal conditions of the extraction yield of flavonoids is as follows in the Stropharia rugoso-annulata fruit body: material and solvent ratios are the material ratio (W/V) of 1:40～70; Extracting solvent is 80%～90%(volume) the food-grade industrial alcohol; The extraction time of 2h～3h; 70 ℃～90 ℃ extraction temperature.Adopt the extraction conditions of optimizing to make the extraction yield of flavonoids in the Stropharia rugoso-annulata fruit body reach 4～9 ‰, improved more than 4 times than the document extracting method.
The preparation method of extract of flavonoids is as follows in the Stropharia rugoso-annulata fruit body of the present invention: make raw material with the Stropharia rugoso-annulata fruit body, adopt the food-grade industrial alcohol to make to extract solvent, Stropharia rugoso-annulata fruit body or its are pulverized material and ratio of solvent (W/V) adding 80%～90%(volume that material press 1:40～70) the food-grade industrial alcohol, refluxing extraction 2～3h, decompression, suction filtration, the concentrated flavonoid compound extract that obtains, carry out freeze drying then, obtain the extract of the flavonoids of Stropharia rugoso-annulata fruit body.With the rutin is contrast, adopts colorimetric method, measures the flavonoids in the Stropharia rugoso-annulata fruiting body extract, and its content accounts for 1～5% of extract gross weight.
A kind of technology that improves flavonoid content in the Stropharia rugoso-annulata fruit body and extraction efficiency thereof of the present invention, the extract of flavonoids can be used for food preservative in the Stropharia rugoso-annulata fruit body that this technology obtained, fresh-keeping as the animal and plant grease, preservation of fishery, really, vegetables are fresh-keeping, all kinds of instant food are fresh-keeping etc.; Also be used for function nutrition development of food such as dietary supplements, food additives; Be used to produce natural anti-oxidation medicine with anti-oxidation characteristics etc.The antioxidation effect of the extract of flavonoids is obvious in the Stropharia rugoso-annulata fruit body, and extraction process is simple, and cost is low, the recovery rate height, can make full use of waste resources such as straw, husk cultivation Stropharia rugoso-annulata, with cheapness, rich in natural resources is as producing the Stropharia rugoso-annulata raw material.
Remarkable advantage of the present invention:
A) the cultivation major ingredient of flavonoids is waste resources such as straw, husk in the raising Stropharia rugoso-annulata fruit body provided by the invention, turns waste into wealth environmental protection.
B) Stropharia rugoso-annulata has stronger degeneration-resistant anti-polygamy, is easy to cultivation, thereby the Stropharia rugoso-annulata aboundresources, the invention provides the approach of a development and use Stropharia rugoso-annulata resource.
C) extracting method operation provided by the invention is few, easy and simple to handle, with short production cycle, and the flavonoid content that extracts in the Stropharia rugoso-annulata fruit body can improve more than 4 times.
Embodiment
Below be several specific embodiment of the present invention, further specify the present invention, but the present invention be not limited only to this.
Flavonoids extracts and assay in the embodiment 1 Stropharia rugoso-annulata fruit body
Flavonoids extraction step in the Stropharia rugoso-annulata fruit body: 60 ℃ of oven dry of Stropharia rugoso-annulata fruit body (dry product), ground 70 eye mesh screens → food-grade industrial alcohol solution circumfluence method extraction → decompression, suction filtration separation of supernatant, constant volume → rotary evaporation concentrates → freeze drying.
Flavonoid content assay method in the Stropharia rugoso-annulata fruit body: with the rutin is standard specimen, adopts colorimetric method, content of flavonoids in the extract of mensuration Stropharia rugoso-annulata flavonoids, the yield of calculating flavonoids.
Computing formula:
The cleaning of Stropharia rugoso-annulata fruit body, oven dry, powder essence with the field results; Accurately take by weighing 15.25g and cross 70 purpose Stropharia rugoso-annulata powder, at material ratio 1:40, the 80%(volume) food-grade industrial alcohol solution concentration, 80 ℃ of temperature, under the extraction conditions of time 2h, the employing circumfluence method is extracted, and the general flavone content of measuring the extract of Stropharia rugoso-annulata flavonoids is 5.98 ‰.Obtain Stropharia rugoso-annulata extract 5.77g after the freeze-drying, Stropharia rugoso-annulata alcohol soluble substance content reaches 37.8%.
Accurately take by weighing Stropharia rugoso-annulata alcohol extract 0.1002g, use the 80%(volume) food-grade industrial alcohol solution water solution, be settled to 5 mL, get the liquid to be measured of 20.04mg/ mL.Accurately measuring Stropharia rugoso-annulata extract 1 mL and insert in the 10mL screw socket tool plug small test tube, is contrast with the rutin, adopts colorimetric method for determining to get absorbance.The flavones content that gets in the flavonoids according to the calibration curve regression equation calculation is 3.02%.
The comparison that embodiment 2 different Stropharia rugoso-annulata culture matrixes are filled a prescription
(1) cultivation
Adopt the whole bedding cultivation mode of the outdoor raw material garden farming of Stropharia rugoso-annulata to plant, select different culture medium prescriptions: be culturing raw material with 100% golden mushroom chaff, 100% husk, 50% husk+50% straw and 33% husk+67% wood chip respectively.
(2) the flavonoids recovery rate calculates
The Stropharia rugoso-annulata fruit body of field results is cleaned, obtain the extract of flavonoids in the Stropharia rugoso-annulata fruit body according to flavonoids extraction step in the embodiment 1 described Stropharia rugoso-annulata fruit body, according to the computing formula of embodiment 1 described Stropharia rugoso-annulata flavonoid content assay method and flavonoids yield, calculate the recovery rate of the Stropharia rugoso-annulata fruit body flavonoids of four kinds of culturing raw material productions:
1) 100% golden mushroom chaff: the flavonoids recovery rate is 4.35 ‰
2) husk wood chip (33% husk and 67% wood chip): the flavonoids recovery rate is 4.71 ‰
3) husk straw (50% husk and 50% straw): the flavonoids recovery rate is 5.62 ‰
4) 100% husk: the flavonoids recovery rate is 5.82 ‰
The Stropharia rugoso-annulata fruit body flavonoids recovery rate that 100% husk prescription is produced is the highest, the economic benefit maximum.
Flavonoids extracts and assay in the embodiment 3 Stropharia rugoso-annulata fruit bodys
Adopting the whole bedding cultivation mode of the outdoor raw material garden farming of Stropharia rugoso-annulata to plant, is that culturing raw material is cultivated with 100% husk, and the Stropharia rugoso-annulata fruit body of field results is cleaned, and 60 ℃ of oven dry ground 70 eye mesh screens.Accurately took by weighing 70 purpose Stropharia rugoso-annulata powder 1g, at material ratio 1:70(W/V), the 90%(volume) the food-grade industrial alcohol, under the extraction conditions that temperature is 90 ℃, circumfluence method is extracted 2h, adopts colorimetric method, is that the general flavone content of reference substance mensuration Stropharia rugoso-annulata extract is 5.16 ‰ with the rutin.
Embodiment 4
Repeat embodiment 3 described cultivation steps, obtain the Stropharia rugoso-annulata fruit body.Accurately took by weighing 70 purpose Stropharia rugoso-annulata powder 1g, at material ratio 1:40(W/V), the 80%(volume) the food-grade industrial alcohol, under the extraction conditions that temperature is 80 ℃, circumfluence method is extracted 3h, adopts colorimetric method, is that the general flavone content of reference substance mensuration Stropharia rugoso-annulata extract is 7.41 ‰ with the rutin.
Embodiment 5
Repeat embodiment 3 described cultivation steps, obtain the Stropharia rugoso-annulata fruit body.Accurately took by weighing 70 purpose Stropharia rugoso-annulata powder 1g, at material ratio 1:50(W/V), the 85%(volume) food-grade industrial alcohol, under the extraction conditions that temperature is 90 ℃, circumfluence method is extracted 2.5h, adopting colorimetric method, is that the general flavone content of reference substance mensuration Stropharia rugoso-annulata extract is 8.23 ‰ with the rutin.
The antioxidation activity of flavonoids is measured in the embodiment 6 Stropharia rugoso-annulata fruit bodys
(1) effect of external removing hydroxy radical: Phen-Fe
2+Oxidizing process
(2) the removing activity of superoxide anion: pyrogallol autoxidation method
(3) the snperoxiaized inhibitory action of unsaturated fatty acid: Fe
2+Bring out the mensuration of antioxidation activity (AOA) in lipovitellinin polyunsaturated fatty acid (PUFA) the peroxidating system
Get one group of Stropharia rugoso-annulata flavonoids extract that recovery rate is the highest in the orthogonal experiment and be used for resistance to oxidation mensuration, measure 0.3mL Stropharia rugoso-annulata flavonoids extract, with 90% food-grade industrial alcohol constant volume in the 50mL volumetric flask, as the liquid to be measured of antioxidation activity experiment.Utilize Phen-Fe
2+Oxidizing process, pyrogallol autoxidation method and Fe
2+The assay method that brings out antioxidation activity (AOA) in lipovitellinin polyunsaturated fatty acid (PUFA) the peroxidating system carries out the mensuration of antioxidation activity to liquid to be measured, the result shows that flavonoids in the Stropharia rugoso-annulata flavonoids extract is 23.07% to the clearance rate of hydroxy radical (OH), to the autoxidizable inhibiting rate of pyrogallol is 26.85%, is 19.26% to the inhibiting rate of lipid peroxidation.
Therefore the Stropharia rugoso-annulata flavonoids has oxidation resistant feature, has removing hydroxy radical (OH), suppresses superoxide anion (O
2-) and suppress the effect of lipid peroxidation.In vitro test is the result show, the Stropharia rugoso-annulata flavonoids has multiple antioxidation.
Claims (2)
1. method that improves flavonoids recovery rate in the Stropharia rugoso-annulata fruit body, it is characterized in that: the prescription of described stropharia rugoso-annulata plantation matrix is 100% husk or 50% husk+50% straw; Adopt the whole bedding cultivation mode of outdoor raw material garden farming to produce plantation, obtain the Stropharia rugoso-annulata fruit body; With food-grade industrial alcohol solution serves as to extract solvent, and described alcohol concentration volume ratio is 80%～90%, and the Stropharia rugoso-annulata fruit body is the material ratio of 1:40～70 with extracting solvent ratios W/V, the extraction time of 2h～3h, 70 ℃～90 ℃ extraction temperature.
2. the method for flavonoids recovery rate in the raising Stropharia rugoso-annulata fruit body according to claim 1 is characterized in that:
Carry out according to following steps
(1) refluxing extraction: it is that material and the ratio of solvent adding volume ratio of 1:40～70 is 80%～90% food-grade industrial alcohol by W/V that Stropharia rugoso-annulata fruit body or its are pulverized material, 70 ℃～90 ℃ refluxing extraction 2～3h, decompression, suction filtration, the concentrated flavonoid compound extract that obtains, carry out freeze drying then, obtain the extract of flavonoids in the Stropharia rugoso-annulata fruit body;
(2) recovery rate calculates: with the rutin is contrast, adopt colorimetric method, volume ratio is that 80%～90% food-grade industrial alcohol is an extract, measures content of flavonoids in the extract of flavonoids in the Stropharia rugoso-annulata fruit body, calculates the yield of flavonoids.
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