CN110882285A - Efficient preparation method of active substances in Phellinus linteus - Google Patents

Abstract

The invention discloses an efficient preparation method of active substances in phellinus igniarius, which comprises the following steps of S1, raw material pretreatment: drying and crushing phellinus igniarius sporocarp to form phellinus igniarius powder; s2, MgO and tetrahydrofuran treatment: adding MgO accounting for 40-60% of the weight of the phellinus igniarius powder and tetrahydrofuran accounting for 20-40 times of the weight of the phellinus igniarius powder into the phellinus igniarius powder, uniformly stirring, placing the phellinus igniarius powder into a reaction kettle, and extracting for 30-60 minutes at the temperature of 200 ℃ and 260 ℃; s3, ultrasonic treatment: cooling to room temperature after reaction, adding water 1-2 times of the volume of the mixture into the mixture, performing ultrasonic treatment at 30-50 deg.C for 10-30 min, and filtering the mixture to obtain filtrate containing Phellinus linteus polyphenol and Phellinus linteus polysaccharide. The filtrate can be further separated to obtain Phellinus linteus polyphenol and Phellinus linteus polysaccharide, respectively. The method can simultaneously prepare the phellinus igniarius polyphenol and the phellinus igniarius polysaccharide, improves the extraction rate of the phellinus igniarius polysaccharide and the phellinus igniarius polyphenol, has simple process, and is suitable for large-scale production.

Description

Efficient preparation method of active substances in Phellinus linteus

technical field

The invention relates to the field of edible and medicinal fungi processing, in particular to an efficient preparation method of active substances in Phellinus linteus.

Background technique

Phellinus linteus is a precious edible and medicinal mushroom, known as "forest gold". Phellinus linteus, as a traditional Chinese medicinal material in my country, was first recorded in "Shen Nong Baicao Jing" and Li Shizhen's "Compendium of Materia Medica". According to research statistics, Phellinus linteus has more than 20 pharmacological functions, including antibacterial, anti-inflammatory, antioxidant, anti-tumor, enhancing immunity, protecting liver and protecting liver, lowering blood sugar, lowering blood lipids, and anti-pneumonia.

The direct consumption of edible and medicinal fungi generally cannot achieve the therapeutic effect. Only by extracting, separating and purifying the active ingredients, can the efficacy be improved and the pharmacological effects be exerted. In the preparation process of active ingredients, extraction is the first and most critical step, and how to release the active ingredients to the greatest extent is of great significance.

Chinese patent document CN108610391A discloses a method for extracting polysaccharides and adenosine from Phellinus linteus fruiting bodies, comprising the following steps: (1) after crushing Phellinus linteus fruiting bodies into Phellinus linteus fruiting body particles, put them into an extraction tank; (2) ) in the extraction tank, add the water of 6-10 times the volume of Phellinus linteus fruit body particles, continuously reflux for 1.5-3 hours, suction filtration, and the filter residue continues to use the water of 4-5 times the volume of the filter residue to continuously reflux for 2-3 hours, pumping filter; (3) mix the filtrate obtained by suction filtration twice in step (2), and obtain 0.3-0.5g/ml Phellinus linteus solution after concentrating; (4) adjust the Phellinus linteus solution in step (3) with sodium hydroxide solution The pH value is to 9-10, the Phellinus linteus solution that adjusts the pH value is sampled in the stainless steel chromatography column that is equipped with macroporous resin, the flow velocity of the control effluent is controlled, and the gained effluent is the thick solution of Phellinus linteus polysaccharide; ( 5) The thick solution of Phellinus linteus polysaccharide in step (4) is adjusted to neutrality with sulfuric acid solution, and through the first ultrafiltration membrane that can filter out macromolecules such as protein, collect the filtrate and pass through the first ultrafiltration membrane that removes small molecule impurities. Two ultrafiltration membranes, the obtained retentate is concentrated under reduced pressure and vacuum to obtain Phellinus linteus polysaccharides; (6) the adsorbate adsorbed in the macroporous resin in step (4) is eluted with ethanol solution, and the obtained eluate is reduced by ethanol solution. The extract is obtained after being concentrated in a vacuum; (7) the extract in step (6) and the silica gel are dry-mixed in a mass ratio of 1:1.2, and the upper chromatographic column is washed with 1-2 column volumes of ethyl acetate first. Then, eluate with ethyl acetate and acetone mixed eluent, collect one fraction per 1500ml, detect in liquid phase, combine the 24th to 29th fractions in the fractions, concentrate under reduced pressure and vacuum, and remove the precipitates that appear. Combined, filtered to collect the precipitate, the filtrate was allowed to stand again, filtered out after crystallization, the filtered precipitate was mixed with the original precipitate, and the mixed precipitate was recrystallized in an acetone-ethanol solution with a volume ratio of 6:4 to obtain a colorless Needle crystal adenosine, suction filtration, and vacuum drying at 50°C in a vacuum desiccator to obtain the finished product of adenosine.

Chinese patent document CN108542922A discloses an efficient preparation method of active ingredients in Phellinus linteus, comprising the following steps: pretreatment of raw materials: (1) drying the Phellinus linteus fruit bodies at 50-60 ° C, pulverizing, and then passing 40-60 Mesh sieving to obtain Phellinus linteus powder; (2) mixing the Phellinus linteus powder obtained in step 1 with deionized water in a ratio of 1:20 to 1:50 by mass and volume, and after mixing evenly, the added mass is 1% to 1% of the mass of Phellinus linteus powder. 2% of cellulase, mixed evenly again, and treated at a temperature of 45°C to 55°C for 1 to 2 hours to obtain a reaction mixture; (3) the reaction mixture obtained in step 2 was added to the extraction kettle for extraction, and the extraction kettle The pressure is 5～20mPa, the extraction temperature is 100℃～200℃, the extraction time is 30～90min, and the extraction finishes to obtain the extract; (4) the extract obtained in step 3 is centrifuged for 10 minutes at the rotating speed of 4500～7000r/min. ~20min, separate the supernatant liquid to obtain the first supernatant liquid; (5) add chlorohexadecylpyridine to the first supernatant liquid, shake well and let stand at room temperature for 24h~36h, at 3500~7000r/ The first precipitate and the second supernatant are obtained by centrifugation at a rotating speed of 1 min for 10 to 15 min; (6) the first precipitate obtained in step 5 is added to the ethanol solution of NaCl to obtain a precipitation mixture, and the precipitation mixture is placed in a shaker Shake for 1 to 1.5 hours, centrifuge at 3500 to 7000 r/min for 10 to 15 minutes, obtain a precipitate after centrifugation, rinse the precipitate with 80% ethanol, and freeze-dry it to obtain highly active Phellinus linteus polysaccharide SP1; (7) Steps 5. The obtained second supernatant is concentrated under reduced pressure and freeze-dried to obtain an extract rich in Phellinus linteus triterpenes and flavonoids.

Chinese patent document CN106491662A discloses a method for simultaneously extracting polysaccharides and flavonoids from Phellinus linteus cultivation waste, comprising the following steps: (1) drying and pulverizing Phellinus Phellinus cultivation waste to obtain Phellinus Phellinus cultivation waste dry powder; Yellow cultivation waste dry powder and alkaline ethanol solution are mixed according to mass ratio of 1:10-1:30; (3) the mixed solution in step (2) is ultrasonically treated for 20-40 minutes, and filtered to obtain filtrate and residue; (4) to step (3) Add deionized water of 8-12 times the weight of the residue to the residue in (3), mix well, adjust the pH value to 4-6 to obtain a mixed solution, add a compound enzyme that accounts for 0.2-0.5% of the weight of the mixed solution, and the compound enzyme is cellulose The enzyme and glucose oxidase are mixed, and enzymolysis is carried out at 45-55°C for 40-80 minutes, and the enzymolyzed solution is centrifuged to separate the supernatant and the precipitate; the compound enzyme is cellulase: glucose oxidase=2-3: The mixture of 1; (5) the filtrate obtained in step (3) is subjected to rotary evaporation to remove ethanol, and ethyl acetate is added for extraction, liquid separation, and the supernatant is concentrated and dried under reduced pressure to obtain Phellinus flavonoids; (6) step (4) The supernatant obtained in ) was concentrated under reduced pressure with a rotary evaporator, and then subjected to alcohol precipitation and centrifugation to obtain a supernatant and a precipitate, and the precipitate was Phellinus linteus polysaccharide.

There is no literature report on the simultaneous extraction of Phellinus Phellinus polyphenols and Phellinus Phellinus polysaccharides in the prior art.

SUMMARY OF THE INVENTION

The inventor believes that this is because the Phellinus linteus fruit body has a high degree of lignification and a complex structure, and the active substances are generally combined with lignin through physical or chemical bonds. The active ingredients are extracted. Therefore, destroying the bond between active substances and lignin by removing lignin is also an effective extraction method.

In view of this, the object of the present invention is to provide a kind of efficient preparation method of active substances in Phellinus linteus, which can simultaneously efficiently extract Phellinus linteus polyphenols and Phellinus linteus polysaccharides, so that the effective components Phellinus flavonoids and Phellinus Phellinus polysaccharides can be simultaneously extracted to the greatest extent. freed.

The technical solutions adopted are:

A kind of efficient preparation method of active substance in Phellinus linteus, comprising the steps:

S1. Raw material pretreatment: dry and pulverize the fruiting bodies of Phellinus linteus to form Phellinus linteus powder;

S2. MgO and tetrahydrofuran processing: add 40%-60% MgO of Phellinus linteus powder weight and 20-40 times tetrahydrofuran of Phellinus linteus powder weight in Phellinus linteus powder, stir evenly and place in the reactor, extract 30% at 200-260 ℃ -60 minutes;

S3. Ultrasonic treatment: after the reaction is completed and lowered to room temperature, water 1-2 times the volume of the mixture is added to the mixture, sonicated at 30-50° C. for 10-30 minutes, and the mixture is filtered to obtain a mixture containing both Phellinus linteus and Phellinus linteus polysaccharide. filtrate.

In this technical solution, on the one hand, the toxicity of tetrahydrofuran is low, and MgO is used as a catalyst to dissolve the lignin in the fruiting bodies of Phellinus Phellinus under the action of the catalyst MgO, so that more Phellinus flavonoids and Phellinus Phellinus polysaccharides are released, thereby improving the extraction rate.

On the other hand, tetrahydrofuran dissolves the lignin of Phellinus linteus fruiting bodies under the catalysis of MgO, and then ultrasonically treats it, so that the effective components Phellinus flavonoids and Phellinus Phellinus polysaccharides are simultaneously released to the greatest extent.

Further, it also includes S4. The filtrate is added with an equal volume of dichloromethane for extraction, and the organic layer is dried to obtain Phellinus linteus. The yield of Phellinus linteus is 2%-4.7%.

Further, it also includes S5. After the water layer is concentrated, anhydrous ethanol is added to make the ethanol concentration reach 70-80% by volume, and the precipitation is collected to obtain Phellinus linteus polysaccharides. The yield of Phellinus linteus polysaccharide is 8.7%-11.2%.

Further, in S2, 50% of MgO by weight of Phellinus linteus powder and 20-40 times of tetrahydrofuran by weight of Phellinus linteus powder are added to Phellinus linteus powder.

Further, in S5, after the water layer is concentrated, anhydrous ethanol is added to make the ethanol concentration reach 75% by volume, and the precipitation is collected to obtain Phellinus linteus polysaccharides.

Further, in S1, superfine pulverization is performed after drying the fruiting bodies of Phellinus linteus at 50-60°C.

Similarly, from the product point of view, the present invention can also provide a mixed solution containing both Phellinus linteus polyphenols and Phellinus linteus polysaccharides, which is prepared by the efficient preparation method described in the above scheme.

Further, the mixed solution is added with an equal volume of dichloromethane for extraction, and the organic layer is dried to obtain Phellinus linteus polyphenols, and the yield of Phellinus linteus polyphenols is 2%-4.7%.

Further, after the water layer is concentrated, anhydrous ethanol is added to make the ethanol concentration reach 70-80% by volume, collect and precipitate to obtain Phellinus linteus polysaccharide, and the yield of Phellinus linteus polysaccharide is 8.7%-11.2%.

The beneficial effects of the present invention are:

The high-efficiency preparation method of the active substances in Phellinus linteus can simultaneously prepare Phellinus linteus polyphenols and Phellinus linteus polysaccharides, and the extraction rate of Phellinus linteus polysaccharides and Phellinus linteus polysaccharides is improved, and the process is simple and suitable for large-scale production.

Detailed ways

The present invention will be described in detail below through specific examples, but the purposes and purposes of these exemplary embodiments are only used to illustrate the present invention, and do not constitute any limitation to the actual protection scope of the present invention, nor do they limit the present invention. The scope of protection is limited to this.

Example 1

An efficient preparation method of active substance in Phellinus linteus, comprising the following steps,

S1. Raw material pretreatment: dry and pulverize the fruiting bodies of Phellinus linteus;

S2. MgO and tetrahydrofuran processing: add 50% of its weight MgO and 20 times the weight of tetrahydrofuran in Phellinus linteus powder, stir well and place in the reactor, and extract at 200 ° C for 30 minutes;

S3. After the reaction is completed and lowered to room temperature, the mixture is added with 1 times its volume of water, sonicated at 30° C. for 10 minutes, and the mixture is filtered;

S4. The filtrate was added with an equal volume of dichloromethane for extraction, and the organic layer was dried to obtain Phellinus linteus, with a yield of 2.1%.

S5. After the water layer is concentrated, anhydrous ethanol is added to make the ethanol concentration reach 75% by volume, and the precipitate is collected to obtain Phellinus linteus polysaccharides. The yield of Phellinus linteus polysaccharide was 8.9%.

Example 2

An efficient preparation method of active substance in Phellinus linteus, comprising the following steps,

S1. Raw material pretreatment: dry and pulverize the fruiting bodies of Phellinus linteus;

S2. MgO and tetrahydrofuran processing: add 50% of its weight MgO and 30 times the weight of tetrahydrofuran in Phellinus linteus powder, stir well and place in a reactor, extract 40 minutes at 220 °C;

D. After the reaction was completed and lowered to room temperature, the mixture was added with 1.5 times its volume of water, sonicated at 40°C for 20 minutes, and the mixture was filtered;

E. The filtrate was added with an equal volume of dichloromethane for extraction, and the organic layer was dried to obtain Phellinus linteus, with a yield of 2.65%.

F. After the water layer is concentrated, anhydrous ethanol is added to make the ethanol concentration reach 75% by volume, and the precipitate is collected to obtain Phellinus linteus polysaccharides. The yield of Phellinus linteus polysaccharide was 9.47%.

Example 3

An efficient preparation method of active substance in Phellinus linteus, comprising the following steps,

S1. Raw material pretreatment: dry and pulverize the fruiting bodies of Phellinus linteus;

S2. MgO and tetrahydrofuran processing: add 50% of its weight MgO and 40 times the weight of tetrahydrofuran in Phellinus linteus powder, stir well and place in the reactor, and extract at 240 ° C for 50 minutes;

S3. After the reaction is completed and lowered to room temperature, 1.8 times the volume of water is added to the mixture, sonicated at 45°C for 25 minutes, and the mixture is filtered;

S4. The filtrate was added with an equal volume of dichloromethane for extraction, and the organic layer was dried to obtain Phellinus linteus, and the yield was 3.46%.

S5. After the water layer is concentrated, anhydrous ethanol is added to make the ethanol concentration reach 75% by volume, and the precipitate is collected to obtain Phellinus linteus polysaccharides. The yield of Phellinus linteus polysaccharide was 10.05%.

Example 4

An efficient preparation method of active substance in Phellinus linteus, comprising the following steps,

S1. Raw material pretreatment: superfine pulverization after drying the fruiting bodies of Phellinus linteus at 55°C;

S2.MgO and tetrahydrofuran processing: add the 50% MgO of its weight and the tetrahydrofuran of 35 times of weight in Phellinus linteus powder, stir and place in the reactor, extract 60 minutes at 250 DEG C;

S3. After the reaction is completed and lowered to room temperature, the mixture is added with 2 times the volume of water, sonicated at 50° C. for 30 minutes, and the mixture is filtered;

S4. The filtrate was added with an equal volume of dichloromethane for extraction, and the organic layer was dried to obtain Phellinus linteus, with a yield of 3.89%.

S5. After the water layer is concentrated, anhydrous ethanol is added to make the ethanol concentration reach 75% by volume, and the precipitate is collected to obtain Phellinus linteus polysaccharides. The yield of Phellinus linteus polysaccharide was 10.54%.

Example 5

An efficient preparation method of active substance in Phellinus linteus, comprising the following steps,

S1. Raw material pretreatment: dry and pulverize the fruiting bodies of Phellinus linteus;

S2. MgO and tetrahydrofuran are processed: add the 50% MgO of its weight and the tetrahydrofuran of 40 times of its weight in Phellinus linteus powder, stir evenly and place in the reactor, and extract 50 minutes at 260 °C;

S3. After the reaction is completed and lowered to room temperature, 1.6 times the volume of water is added to the mixture, sonicated at 45° C. for 25 minutes, and the mixture is filtered;

S4. The filtrate was added with an equal volume of dichloromethane for extraction, and the organic layer was dried to obtain Phellinus linteus, and the yield was 3.55%.

S5. After the water layer is concentrated, anhydrous ethanol is added to make the ethanol concentration reach 75% by volume, and the precipitate is collected to obtain Phellinus linteus polysaccharides. The yield of Phellinus linteus polysaccharide was 9.98%.

The series of detailed descriptions listed above are only specific descriptions for the feasible embodiments of the present invention, and they are not intended to limit the protection scope of the present invention. Changes should all be included within the protection scope of the present invention.

Claims (9)

1. an efficient preparation method of active material in Phellinus linteus, is characterized in that, comprises the steps: S1. Raw material pretreatment: dry and pulverize the fruiting bodies of Phellinus linteus to form Phellinus linteus powder; S2. MgO and tetrahydrofuran processing: add 40%-60% MgO of Phellinus linteus powder weight and 20-40 times tetrahydrofuran of Phellinus linteus powder weight in Phellinus linteus powder, stir evenly and place in the reactor, extract 30% at 200-260 ℃ -60 minutes; S3. Ultrasonic treatment: after the reaction is completed and lowered to room temperature, water 1-2 times the volume of the mixture is added to the mixture, sonicated at 30-50° C. for 10-30 minutes, and the mixture is filtered to obtain a mixture containing both Phellinus linteus and Phellinus linteus polysaccharide. filtrate. 2. the efficient preparation method of active material in Phellinus linteus according to claim 1, is characterized in that, also comprises S4. Filtrate adds equal volume of dichloromethane to extract, and organic layer obtains Phellinus linteus polyphenol after drying. 3. the efficient preparation method of active material in Phellinus linteus according to claim 2, is characterized in that, also comprises S5. after the water layer is concentrated, adding dehydrated alcohol to make ethanol concentration reach volume fraction is 70-80%, collect precipitation to obtain 70-80%. Phellinus polysaccharide. 4. the efficient preparation method of active substance in Phellinus linteus according to claim 3, is characterized in that, in S2, in Phellinus linteus powder, add the 50% MgO of Phellinus linteus powder weight and the tetrahydrofuran of 20-40 times of Phellinus linteus powder weight . 5. the efficient preparation method of active substance in Phellinus linteus according to claim 3, is characterized in that, in S5, adding dehydrated alcohol after the water layer is concentrated and making ethanol concentration reach volume fraction is 75%, collect and precipitate to obtain Phellinus linteus polysaccharides . 6. The efficient preparation method of active material in Phellinus linteus according to claim 3, is characterized in that, in S1, superfine pulverization is carried out after drying the fruiting bodies of Phellinus linteus at 50-60 DEG C. 7. A mixed solution containing Phellinus linteus and Phellinus linteus polysaccharides simultaneously, is characterized in that, it is prepared by the efficient preparation method described in claim 1. 8. the mixed solution that contains Phellinus linteus and Phellinus linteus polysaccharide simultaneously according to claim 7, is characterized in that, adds equal volume of dichloromethane and extracts, and organic layer obtains Phellinus linteus polyphenols after drying, Phellinus linteus The yield of polyphenols is 2%-4.7%. 9. the mixed solution that contains Phellinus linteus and Phellinus linteus polysaccharide simultaneously according to claim 7, is characterized in that, adding dehydrated alcohol after the water layer is concentrated and making the ethanol concentration reach volume fraction is 70-80%, collects the precipitation to obtain 70-80%. Phellinus linteus polysaccharide, the yield of Phellinus linteus polysaccharide is 8.7%-11.2%.