CN107158044A - A kind of method for integrated extraction of ganoderma lucidum liquid extract - Google Patents
A kind of method for integrated extraction of ganoderma lucidum liquid extract Download PDFInfo
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- 240000008397 Ganoderma lucidum Species 0.000 title claims abstract description 63
- 235000001637 Ganoderma lucidum Nutrition 0.000 title claims abstract description 63
- 239000000284 extract Substances 0.000 title claims abstract description 56
- 239000007788 liquid Substances 0.000 title claims abstract description 46
- 238000000605 extraction Methods 0.000 title claims abstract description 32
- 238000000034 method Methods 0.000 title claims abstract description 30
- 108090000790 Enzymes Proteins 0.000 claims abstract description 31
- 102000004190 Enzymes Human genes 0.000 claims abstract description 31
- 229940088598 enzyme Drugs 0.000 claims abstract description 31
- 230000002195 synergetic effect Effects 0.000 claims abstract description 12
- 108010059892 Cellulase Proteins 0.000 claims abstract description 11
- 229940106157 cellulase Drugs 0.000 claims abstract description 11
- 108090000526 Papain Proteins 0.000 claims abstract description 10
- 239000004365 Protease Substances 0.000 claims abstract description 10
- 229940055729 papain Drugs 0.000 claims abstract description 10
- 235000019834 papain Nutrition 0.000 claims abstract description 10
- 230000007071 enzymatic hydrolysis Effects 0.000 claims abstract description 5
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 claims abstract description 5
- 239000002994 raw material Substances 0.000 claims abstract description 3
- 238000005516 engineering process Methods 0.000 claims description 7
- 239000000203 mixture Substances 0.000 claims description 3
- 230000007062 hydrolysis Effects 0.000 abstract description 4
- 238000006460 hydrolysis reaction Methods 0.000 abstract description 4
- 238000002525 ultrasonication Methods 0.000 abstract 1
- 238000012360 testing method Methods 0.000 description 23
- 238000002474 experimental method Methods 0.000 description 15
- 238000011084 recovery Methods 0.000 description 11
- 241000222336 Ganoderma Species 0.000 description 7
- 230000000694 effects Effects 0.000 description 7
- 238000004458 analytical method Methods 0.000 description 6
- 230000008569 process Effects 0.000 description 6
- 238000011160 research Methods 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 239000004480 active ingredient Substances 0.000 description 5
- 238000004519 manufacturing process Methods 0.000 description 5
- 239000000843 powder Substances 0.000 description 5
- 238000001027 hydrothermal synthesis Methods 0.000 description 4
- 229920001282 polysaccharide Polymers 0.000 description 4
- 239000005017 polysaccharide Substances 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 102000003712 Complement factor B Human genes 0.000 description 3
- 108090000056 Complement factor B Proteins 0.000 description 3
- 230000000259 anti-tumor effect Effects 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 150000004676 glycans Chemical class 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- -1 ucleosides Chemical class 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 238000000540 analysis of variance Methods 0.000 description 2
- 239000003963 antioxidant agent Substances 0.000 description 2
- 230000003078 antioxidant effect Effects 0.000 description 2
- 235000006708 antioxidants Nutrition 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 210000000170 cell membrane Anatomy 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 235000013399 edible fruits Nutrition 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 210000004185 liver Anatomy 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- BHZOKUMUHVTPBX-UHFFFAOYSA-M sodium acetic acid acetate Chemical compound [Na+].CC(O)=O.CC([O-])=O BHZOKUMUHVTPBX-UHFFFAOYSA-M 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 150000003648 triterpenes Chemical class 0.000 description 2
- 238000012795 verification Methods 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 244000025254 Cannabis sativa Species 0.000 description 1
- 241000282994 Cervidae Species 0.000 description 1
- QXNVGIXVLWOKEQ-UHFFFAOYSA-N Disodium Chemical compound [Na][Na] QXNVGIXVLWOKEQ-UHFFFAOYSA-N 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 244000000231 Sesamum indicum Species 0.000 description 1
- 235000003434 Sesamum indicum Nutrition 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 229930182558 Sterol Natural products 0.000 description 1
- 230000001133 acceleration Effects 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 229930013930 alkaloid Natural products 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000003712 anti-aging effect Effects 0.000 description 1
- 210000003056 antler Anatomy 0.000 description 1
- 206010003246 arthritis Diseases 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 238000000205 computational method Methods 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 235000008504 concentrate Nutrition 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- HHEAADYXPMHMCT-UHFFFAOYSA-N dpph Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1[N]N(C=1C=CC=CC=1)C1=CC=CC=C1 HHEAADYXPMHMCT-UHFFFAOYSA-N 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 150000002240 furans Chemical class 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 238000003760 magnetic stirring Methods 0.000 description 1
- 229910000403 monosodium phosphate Inorganic materials 0.000 description 1
- 235000019799 monosodium phosphate Nutrition 0.000 description 1
- 238000000465 moulding Methods 0.000 description 1
- 229920001277 pectin Polymers 0.000 description 1
- 239000001814 pectin Substances 0.000 description 1
- 235000010987 pectin Nutrition 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-N phosphoric acid Substances OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000002000 scavenging effect Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000007873 sieving Methods 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 239000007974 sodium acetate buffer Substances 0.000 description 1
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 description 1
- 208000010110 spontaneous platelet aggregation Diseases 0.000 description 1
- 150000003432 sterols Chemical class 0.000 description 1
- 235000003702 sterols Nutrition 0.000 description 1
- 239000006188 syrup Substances 0.000 description 1
- 235000020357 syrup Nutrition 0.000 description 1
- 238000005199 ultracentrifugation Methods 0.000 description 1
- 238000002137 ultrasound extraction Methods 0.000 description 1
Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/06—Fungi, e.g. yeasts
- A61K36/07—Basidiomycota, e.g. Cryptococcus
- A61K36/074—Ganoderma
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
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Abstract
The invention discloses a kind of method for integrated extraction of ganoderma lucidum liquid extract, it is, using Juncao Ganoderma lucidum as raw material, to be extracted using conbined enzymolysis synergistic supersonic wave method, obtains the ganoderma lucidum liquid extract；Wherein, complex enzyme used is cellulase 2wt%, papain 0.8wt%, pectase 3wt%, and enzymatic hydrolysis condition is：60 DEG C of hydrolysis temperature, pH value is 5.5, and enzymolysis time is 80min, and ultrasonication condition is：Ultrasonic power 300W, 35 DEG C of temperature, ultrasonic time 15min.Compared with conventional hydrothermal extraction method, the yield for extracting liquid extract using complex enzyme hydrolysis synergistic supersonic wave method of the present invention can improve 16.71 times, and can effectively shorten extraction time.
Description
Technical field
The invention belongs to Chinese medicine extractive technique field, and in particular to a kind of integrated extraction side of JUNCAO Ganoderma lucidum liquid extract
Method.
Background technology
Ganoderma lucidum is《Sheng Nong's herbal classic》" medicine-feeding " worthy of the name recorded, is considered as lucky long-lived symbol, enjoys
The good reputation of " celestial grass ".And Juncao Ganoderma lucidum is higher than the active ingredient of general ganoderma lucidum by 3%, security is also higher, immune with improving
The health-care effects such as power, improvement sleep, antitumor, liver protection, auxiliary treatment arthritis.It is isolated from ganoderma lucidum at present：Triterpene
Class compound, polysaccharide, ucleosides, sterols, alkaloids, furan derivatives, amino acid polypeptide class, inorganic elements and fat
The Multiple components such as acid.Polysaccharide compound is the important chemical composition contained by ganoderma lucidum, is one of principle active component of ganoderma lucidum,
Because its is antitumor, immunological regulation, the anti-oxidant and anti-ageing effect of waiting for a long time extremely people pay close attention to.Modern scientific research shows, in ganoderma lucidum
Triterpenes components be also one of its principle active component, with antitumor, liver protection, it is anti-oxidant, suppress platelet aggregation, it is anti-true
The multiple pharmacological effects such as bacterium.
Ganoderma lucidum is shredded, extracts, concentrate by certain technological parameter, be evaporated, what is obtained is in cervine paste, is referred to as
Ganoderma lucidum liquid extract.All production ganoderma lucidum tablets, capsule, electuary or syrup are both needed to prepare glossy ganoderma extract first, its quality it is excellent
The bad quality to Lingzhu ' moulding process and finished dosage form plays very important effect.By improveing liquid extract extraction side
Method, improves the recovery rate of active ingredient in liquid extract, will can effectively improve the production efficiency of ganoderma lucidum liquid extract in industry, reduction life
Produce cost, the quality and stability of medicine are improved, so as to accelerate the modernization of ganoderma lucidum class product.
Research in terms of log glossy ganoderma is a lot, has substantial amounts of document to look into, technology and method all comparative maturities, but right
In the research of JUNCAO Ganoderma lucidum be not a lot, and now JUNCAO Ganoderma lucidum by the advantage such as the low, quality of finished product good of its production cost, by
Gradually in whole nation popularization, this kind of JUNCAO Ganoderma lucidum studies the important kind in this field by ganoderma lucidum is increasingly becoming.Particularly, at present
Research in terms of JUNCAO Ganoderma lucidum liquid extract is blank, therefore, and the research in terms of development JUNCAO Ganoderma lucidum liquid extract especially has
Meaning.
The present invention chooses Juncao Ganoderma lucidum as the object of Enzymatic Extraction, and the cell membrane knot of ganoderma lucidum is destroyed by complex enzyme
Structure, to improve the extraction efficiency of effective component of glossy ganoderma；And in extraction process, judder, the stirring produced using ultrasonic wave
Effect and strong cavitation effect, can accelerate material active ingredient to enter solvent, and improve the abundant of enzyme and effective extract
Contact, shortens extraction time, improves recovery rate, and the process modification for production ganoderma lucidum liquid extract provides theoretical foundation.And be compound
The application of enzyme and ultrasonic synergistic technology in terms of Chinese medicine preparation provides foundation, helps to better control over the production of ganoderma lucidum liquid extract
Amount and quality.
The content of the invention
It is an object of the invention to provide a kind of method for integrated extraction of ganoderma lucidum liquid extract, it can significantly improve Juncao deer horn
Ganoderma lucidum liquid extract yield, and effectively shorten extraction time.
To achieve the above object, the present invention is adopted the following technical scheme that：
A kind of method for integrated extraction of ganoderma lucidum liquid extract, it is, using Juncao Ganoderma lucidum as raw material, to be cooperateed with using conbined enzymolysis
Supercritical ultrasonics technology is extracted, and obtains the ganoderma lucidum liquid extract.It specifically includes following steps：
1）Juncao Ganoderma lucidum is cut into sheet, 65 DEG C of drying, grinding and sievings obtain the glossy ganoderma powder that fineness is 80 mesh；
2）Acetic acid-sodium acetate buffer solution is added in gained glossy ganoderma powder and complex enzyme is digested；
3）After enzymolysis terminates, it is placed into 10min in 100 DEG C of water-baths, makes enzyme-deactivating, room temperature is subsequently cooled to, and surpassed
Sound wave is extracted；
4）Gained extract solution is concentrated under reduced pressure through filtering, is freeze-dried, and the ganoderma lucidum liquid extract is made.
Step 2）The addition of middle complex enzyme is cellulase 2wt%, papain 0.8wt%, pectase 3wt%；It is compound
The condition of enzymolysis is：60 DEG C of hydrolysis temperature, pH value is 5.5, and enzymolysis time is 80min.
Step 3）The condition of middle ultrasonic wave extraction is：Ultrasonic power 300W, 35 DEG C of temperature, ultrasonic time 15min.
The remarkable advantage of the present invention is：The yield that conventional hydrothermal method extracts Juncao Ganoderma lucidum liquid extract is 2.71%, and
The yield that complex enzyme hydrolysis synergistic supersonic wave method of the present invention extracts Juncao Ganoderma lucidum liquid extract is 45.29%, i.e., using present invention side
Method extracts Juncao Ganoderma lucidum, and the more conventional method of its liquid extract yield can be made to improve 16.71 times, and when effectively can shorten extraction
Between.And ultrasonic extraction method is used, its extraction process is without high temperature, it is to avoid because high temperature is to the chemical molecular structure such as GL-B
Destruction, and no matter the Scavenging activity of polyoses content or DPPH free radicals, reducing power, ORAC values are obviously higher than conventional method
Gained liquid extract.
Embodiment
1. materials and methods
1.1 material
1.1.1 research object
Juncao Ganoderma lucidum, is purchased from University Of Agriculture and Forestry In Fujian.
1.1.2 key instrument
Ultracentrifugation beveller（Germany, RetschZM200）；Numerical control ultrasonic cleaner（KQ-500DE types, city of Kunshan's ultrasonic instrument
Co., Ltd）, ultraviolet-uisible spectrophotometer（Beijing Pu Xi all purpose instruments Co., Ltd）, pH meter（Thunder magnetic PHS-3C, Xiamen essence
Do the skill Co., Ltd Foochow that makes the science and technology prosperous）, magnetic stirring apparatus（IKA-Werke GmbH and Co.KG）, freeze drier（Beijing
Bo Yikang laboratory apparatus Co., Ltd）, thermostat water bath（Shanghai Yarong Biochemical Instrument Plant）, rotary evaporator RE52CS（Shanghai
Ya Rong biochemical instruments factory）, SHZ-D (III) circulating water type vavuum pump（Gong Yi cities Yu Hua instruments Co., Ltd）, digital display constant temperature
Water-bath HH-6（Guo Hua Electrical Appliances Co., Ltd）.
1.1.3 main agents
Cellulase 400（μ/mg）, papain（1000μ/mg）, pectase（500μ/mg）, acetic acid, sodium acetate, phosphoric acid hydrogen
Disodium, sodium dihydrogen phosphate.
1.2 method
1.2.1 the extraction of ganoderma lucidum liquid extract
Juncao Antler Mythic Fungus sporophore is cut into sheet, 65 DEG C of drying are crushed, are screened into 80 mesh, then weigh spirit obtained by 5.0 g
Sesame powder, adds the buffer solution of certain pH value（Acetic acid-sodium acetate pH of cushioning fluid is 4.6-5.5）100.0 mL, and add appropriate
Enzyme, fully mix, constant temperature enzymolysis.After reaction terminates, 10min in 100 DEG C of water-baths is placed on, makes enzyme-deactivating, then cools down
To room temperature, extract solution is concentrated under reduced pressure into 10mL or so through filtering by rotary evaporator, and freeze-drying to constant weight calculates gained
The recovery rate of ganoderma lucidum liquid extract.Its computational methods is as follows：
Ganoderma lucidum liquid extract recovery rate/%=×100%。
1.2.2 complex enzyme consumption proportion Orthogonal Experiment and Design
Under 60 min, 50 DEG C, the enzymatic hydrolysis condition that pH value is 5, using the consumption of cellulase, papain and pectase as just
Hand over 3 factors of experiment.Each factor respectively sets 4 levels, carries out L16(43) orthogonal test（Table 1）.
The complex enzyme consumption proportion orthogonal test factor level table of table 1
1.2.3 complex enzyme zymohydrolysis condition Orthogonal Experiment and Design
According to best complex enzyme dosage proportioning test result, the time is carried as investigation factor, each factor using temperature, pH value, enzyme
Respectively 3 levels of setting, carry out L9（33）Orthogonal test（Table 2）.
The complex enzyme zymohydrolysis condition orthogonal test factor level table of table 2
1.2.4 Ultrasonic Conditions orthogonal
According to best complex enzyme dosage proportioning test and optimum enzymolysis condition result of the test, using time, temperature, power as examining
Factor is examined, each factor respectively sets 3 levels, carries out L9（33）Orthogonal test（Table 3）.
The Ultrasonic Conditions orthogonal experiment factor level table of table 3
1.2.5 complex enzyme-supersonic synergic extraction method and the comparison of traditional water bath extraction
8 parts of 5.0 g glossy ganoderma powders are weighed, wherein 4 parts are extracted optimum process condition according to complex enzyme synergistic supersonic wave and extracted, separately
Outer 4 parts be separately added into 100mL concentration 40% ethanol solution, 60 DEG C of water-baths extract 80min.
2 results and analysis
2.1.1 complex enzyme consumption proportion orthogonal test
The complex enzyme consumption proportion orthogonal experiments of table 4
Gained orthogonal experiments can intuitively be found out from table 4, and the K4 of A factors is higher than K1, K2, K3, it is known that factor A 4 water
Flat to be better than other levels, the K4 of B factors is higher than K1, K2, K3, it is known that factor B 4 levels are better than other levels, and the K3 of C factors is high
In K1, K2, K4, it is known that factor C 3 levels are better than other levels.It is thus determined that the complex enzyme of Juncao Ganoderma lucidum liquid extract is used
Amount proportioning is A4B4C3, i.e. cellulase（A）2.0%, papain（B）0.8%, pectase（C）3.0% combination, 60min,
50 DEG C, pH value is under 4.6 enzymatic hydrolysis condition, the average recoveries of ganoderma lucidum liquid extract is 56.81%.
Extreme difference R reflects the amplitude of variation of K values when same factor takes varying level, is varying level correspondence under same factor
Maxima and minima difference.Amplitude of variation is bigger, illustrates that the influence of the factors on test result is bigger, more important.It is fine
The plain enzyme of dimension（A）, papain（B）, pectase（C）Extreme difference R values be respectively 17.64,15.84,13.45, it is possible thereby to push away
Export cellulase（A）Influence to result is maximum, pectase（C）Influence to result is minimum.Influence of the three to result from
Arrive greatly and small be followed successively by cellulase（A）, papain（B）, pectase（C）, wherein, cellulase（A）It is most important, pectin
Enzyme（C）It is least important.Seen by analyze data directly perceived, factor A, B, C G values are all higher than 1.5, i.e., each factor is principal element,
Choose varying level and generation considerable influence is extracted to JUNCAO Ganoderma lucidum liquid extract, influence degree is followed successively by A, B, C.
In order to determine whether whether test error and experimental condition influence result of the test, by orthogonal experiment data progress side
Difference is analysed and significance test, finds out the principal element played a leading role in these factors.From the orthogonal experiment variance analysis knot of table 5
Knowable to fruit, cellulase（A）, papain（B）, pectase（C）Influence to Juncao Ganoderma lucidum liquid extract recovery rate is all
There are conspicuousness, i.e. cellulase（A）, papain（B）, pectase（C）All it is dominant factor.
The complex enzyme consumption proportion of table 5 tests the results of analysis of variance
2.1.2 extraction process verification experimental verification
The best of breed A4B4C3 derived by orthogonal test, in triplicate.As a result being averaged for its ganoderma lucidum liquid extract recovery rate is shown
It is worth for 56.81%, RSD=6.65%, is maximum.
2.2 complex enzyme zymohydrolysis condition orthogonal tests
The complex enzyme zymohydrolysis condition orthogonal experiments of table 6
Can intuitively it find out from the orthogonal experiments of table 6, the k3 of A factors is higher than k1, k2, it is known that factor A 3 levels are better than other
Level, the k2 of B factors is higher than k1, k3, it is known that factor B 2 levels are better than other levels, and the k2 of C factors is higher than k1, k3, derived
Factor C 2 levels are better than other levels.It is thus determined that the enzymatic hydrolysis condition of Juncao Ganoderma lucidum liquid extract is A3B2C2, that is, digest
Temperature（A）For 60 DEG C, pH value（B）5.5, time（C）80 minutes.Temperature（A）、pH（B）, the time（C）R values be respectively 3.01,
3.30,11.83, it can thus be appreciated that the time（C）Influence to result is maximum, temperature（A）Influence to result is minimum.Factor primary and secondary is arranged
Row order is C>B>C, i.e., most important factors time 80min（C）、pH5.5（B）, 40 DEG C of temperature（A).By analyze data directly perceived
See, the G values of factor A, B are respectively less than 1.5, i.e. temperature（A）, pH value（B) factor is secondary cause, time（C）G values be more than
1.5, i.e. time（C）Factor is key factor.
In order to determine whether whether test error and experimental condition influence result of the test, by orthogonal experiment data progress side
Difference is analysed and significance test, finds out the principal element played a leading role in these factors.From the orthogonal experiment variance analysis knot of table 7
Knowable to fruit, temperature（A）、pH（B）, the time（C）Influence to Juncao Ganoderma lucidum liquid extract recovery rate is all without conspicuousness, temperature
（A）、pH（B）, the time（C）It is not dominant factor.
The complex enzyme zymohydrolysis conditional variance analysis result of table 7
2.3 Ultrasonic Conditions orthogonal experiments
The Ultrasonic Conditions Orthogonal experiment results of table 8
Can intuitively it find out from the Orthogonal experiment results of table 8, the k2 of A factors is higher than k1, k3, it is known that factor A 2 levels are better than other
Level, the k3 of B factors is higher than k1, k2, it is known that factor B 3 levels are better than other levels, and the k1 of C factors is higher than k2, k3, derived
Factor C 1 level is better than other levels.It is thus determined that the Ultrasonic Conditions of Juncao Ganoderma lucidum liquid extract are A2B3C1, that is, surpass
The sound time（A）For 15min, power（B) it is 500W, temperature is 35 DEG C.From range analysis, factor primary and secondary puts in order as C>
A>B, i.e. most important factor are temperature（C）, secondly it is the time（A）, power（B）.Seen by analyze data directly perceived, factor A, B, C G
Value is respectively less than 1.5, i.e. time（A）, power（B), temperature（C）Factor is secondary cause.
In order to determine whether whether test error and experimental condition influence result of the test, orthogonal test is subjected to variance point
Analysis, finds out the principal element played a leading role in each factor.From the results of analysis of variance of table 9, A, B, C level of signifiance are all big
In 0.05, illustrate the time（A）, power（B), temperature（C）Influence to JUNCAO Ganoderma lucidum liquid extract recovery rate is not notable, that is,
Say the time（A）, power（B), temperature（C）It is not to influence the dominant factor of JUNCAO Ganoderma lucidum liquid extract recovery rate（P ＞ 0.05）.
The supersonic test variance experimental result of table 9
2.4 complex enzymes-supersonic synergic extraction method is compared with traditional water bath extraction
In order to investigate complex enzyme-supersonic synergic extraction method and influence of the conventional hydrothermal extraction method to ganoderma lucidum liquid extract recovery rate, claim
8 parts of 5.0 g glossy ganoderma powders are taken, wherein 4 parts are extracted optimum process condition according to complex enzyme synergistic supersonic wave and are extracted, 4 parts in addition
100mL water is separately added into, 92 DEG C of hydro-thermals are extracted 11 hours.
As a result show, use the average yield of Juncao Ganoderma lucidum liquid extract of conventional hydrothermal method extraction for 2.71%, wherein
The average content of polysaccharide is 0.23g, and uses being averaged for the Juncao Ganoderma lucidum liquid extract that complex enzyme synergistic supersonic wave method extracts
Yield is 45.29%, and wherein the average content of polysaccharide is the Juncao Ganoderma lucidum that 2.95g, i.e. complex enzyme synergistic supersonic wave method are extracted
The yield of liquid extract is 16.71 times that conventional hydrothermal method is extracted, and polyoses content is that conventional hydrothermal method is extracted in gained liquid extract
12.83 times.This be probably because ultrasonic assistant extractive technique is as a kind of new separation technology, with stirring action, compared with
Strong resonance effects and high acceleration and cavitation effect for producing etc., and mass transport process is strengthened, cause active ingredient to enter
Solvent is accelerated, and improves extraction efficiency, and it will not weaken the activity of organic principle because producing higher temperature.Meanwhile, ganoderma lucidum
Cell quality is hard, and cell membrane is thicker, and the resistance to mass tranfer in extraction process passes through and digests destruction eucaryotic cell structure than larger
To strengthen the mass transport process of active ingredient.
The foregoing is only presently preferred embodiments of the present invention, all equivalent changes done according to scope of the present invention patent with
Modification, should all belong to the covering scope of the present invention.
Claims (4)
1. a kind of method for integrated extraction of ganoderma lucidum liquid extract, it is characterised in that：Using Juncao Ganoderma lucidum as raw material, using complex enzyme
Solution synergistic supersonic wave method is extracted, and obtains the ganoderma lucidum liquid extract.
2. the method for integrated extraction of ganoderma lucidum liquid extract according to claim 1, it is characterised in that：The composition and its use of complex enzyme
Measure as cellulase 2wt%, papain 0.8wt%, pectase 3wt%.
3. the method for integrated extraction of ganoderma lucidum liquid extract according to claim 1, it is characterised in that：Enzymatic hydrolysis condition is：Enzymolysis temperature
60 DEG C of degree, pH value is 5.5, and enzymolysis time is 80min.
4. the method for integrated extraction of ganoderma lucidum liquid extract according to claim 1, it is characterised in that：The treatment conditions of supercritical ultrasonics technology
For：Ultrasonic power 300W, 35 DEG C of temperature, ultrasonic time 15min.
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Effective date of registration: 20231212 Address after: No. 18 Shanghai Road, Baohe Economic Development Zone, Hefei City, Anhui Province, 230601 Patentee after: Anhui huairentang Pharmaceutical Co.,Ltd. Address before: No. 15, Cangshan District, Fujian, Fuzhou, Fujian Patentee before: FUJIAN AGRICULTURE AND FORESTRY University |