CN107412546A - A kind of method of Lilium lancifo1ium Thunb extraction saponin(e - Google Patents
A kind of method of Lilium lancifo1ium Thunb extraction saponin(e Download PDFInfo
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- CN107412546A CN107412546A CN201710573146.9A CN201710573146A CN107412546A CN 107412546 A CN107412546 A CN 107412546A CN 201710573146 A CN201710573146 A CN 201710573146A CN 107412546 A CN107412546 A CN 107412546A
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- lilium
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- 241000234435 Lilium Species 0.000 title claims abstract description 81
- 238000000605 extraction Methods 0.000 title claims abstract description 40
- 238000000034 method Methods 0.000 title claims abstract description 37
- 229930182490 saponin Natural products 0.000 title claims abstract description 31
- 150000007949 saponins Chemical class 0.000 title claims abstract description 31
- 239000001397 quillaja saponaria molina bark Substances 0.000 title claims abstract description 30
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 118
- 239000007787 solid Substances 0.000 claims abstract description 44
- 239000006228 supernatant Substances 0.000 claims abstract description 40
- 239000007788 liquid Substances 0.000 claims abstract description 34
- 239000002994 raw material Substances 0.000 claims abstract description 28
- 241001047198 Scomberomorus semifasciatus Species 0.000 claims abstract description 14
- 235000012467 brownies Nutrition 0.000 claims abstract description 14
- 239000003480 eluent Substances 0.000 claims abstract description 11
- 239000011347 resin Substances 0.000 claims abstract description 10
- 229920005989 resin Polymers 0.000 claims abstract description 10
- 239000000203 mixture Substances 0.000 claims description 54
- 235000015097 nutrients Nutrition 0.000 claims description 19
- 241000894006 Bacteria Species 0.000 claims description 16
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 16
- 239000000284 extract Substances 0.000 claims description 16
- 239000000725 suspension Substances 0.000 claims description 16
- 230000000717 retained effect Effects 0.000 claims description 12
- 239000000047 product Substances 0.000 claims description 11
- 241000223261 Trichoderma viride Species 0.000 claims description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 10
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 9
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 9
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 9
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 9
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims description 9
- 238000004140 cleaning Methods 0.000 claims description 9
- 239000008103 glucose Substances 0.000 claims description 9
- 238000012545 processing Methods 0.000 claims description 9
- 239000002689 soil Substances 0.000 claims description 9
- 150000001413 amino acids Chemical class 0.000 claims description 8
- 230000001580 bacterial effect Effects 0.000 claims description 8
- ORTQZVOHEJQUHG-UHFFFAOYSA-L copper(II) chloride Chemical compound Cl[Cu]Cl ORTQZVOHEJQUHG-UHFFFAOYSA-L 0.000 claims description 8
- FBAFATDZDUQKNH-UHFFFAOYSA-M iron chloride Chemical compound [Cl-].[Fe] FBAFATDZDUQKNH-UHFFFAOYSA-M 0.000 claims description 8
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 8
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 8
- 238000002525 ultrasonication Methods 0.000 claims description 8
- 150000001875 compounds Chemical class 0.000 claims 1
- 238000011084 recovery Methods 0.000 abstract description 14
- 239000000463 material Substances 0.000 abstract description 5
- 241000196324 Embryophyta Species 0.000 abstract description 3
- 239000011343 solid material Substances 0.000 abstract description 2
- 235000017709 saponins Nutrition 0.000 description 22
- 230000000052 comparative effect Effects 0.000 description 9
- 238000010828 elution Methods 0.000 description 7
- 230000000694 effects Effects 0.000 description 5
- 239000004615 ingredient Substances 0.000 description 5
- 235000016709 nutrition Nutrition 0.000 description 5
- 108090000790 Enzymes Proteins 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
- 229940088598 enzyme Drugs 0.000 description 2
- 230000003834 intracellular effect Effects 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000002703 mutagenesis Methods 0.000 description 2
- 231100000350 mutagenesis Toxicity 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 230000002936 tranquilizing effect Effects 0.000 description 2
- 108010059892 Cellulase Proteins 0.000 description 1
- 206010011224 Cough Diseases 0.000 description 1
- 206010057315 Daydreaming Diseases 0.000 description 1
- MKYBYDHXWVHEJW-UHFFFAOYSA-N N-[1-oxo-1-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)propan-2-yl]-2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidine-5-carboxamide Chemical compound O=C(C(C)NC(=O)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F)N1CC2=C(CC1)NN=N2 MKYBYDHXWVHEJW-UHFFFAOYSA-N 0.000 description 1
- 206010033557 Palpitations Diseases 0.000 description 1
- 206010036790 Productive cough Diseases 0.000 description 1
- 208000037656 Respiratory Sounds Diseases 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 241000377209 Unicorn Species 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 230000000840 anti-viral effect Effects 0.000 description 1
- 239000010692 aromatic oil Substances 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 229940106157 cellulase Drugs 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- 208000013116 chronic cough Diseases 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 238000004945 emulsification Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 229930182470 glycoside Natural products 0.000 description 1
- 150000002338 glycosides Chemical class 0.000 description 1
- 235000013402 health food Nutrition 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 230000002218 hypoglycaemic effect Effects 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 229910001410 inorganic ion Inorganic materials 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 150000002576 ketones Chemical class 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 230000003204 osmotic effect Effects 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 230000001869 rapid Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000010008 shearing Methods 0.000 description 1
- 239000000344 soap Substances 0.000 description 1
- 238000010129 solution processing Methods 0.000 description 1
- 230000002557 soporific effect Effects 0.000 description 1
- 235000013599 spices Nutrition 0.000 description 1
- 208000024794 sputum Diseases 0.000 description 1
- 210000003802 sputum Anatomy 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 230000009466 transformation Effects 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
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/896—Liliaceae (Lily family), e.g. daylily, plantain lily, Hyacinth or narcissus
- A61K36/8967—Lilium, e.g. tiger lily or Easter lily
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- 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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- 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
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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- 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
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
-
- 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
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
-
- 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
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/55—Liquid-liquid separation; Phase separation
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
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Abstract
The present invention provides a kind of method of Lilium lancifo1ium Thunb extraction saponin(e, belongs to field of plant extraction, comprises the following steps：Raw material is used as using full Lilium lancifo1ium Thunb bulb first, Boli scale after being cleaned, gained scale is put in ethanol solution under specified conditions, and extracted in ultrasonic environment, solid content and supernatant are centrifuged after extraction, gained solid material is extracted by the way of microwave abstracting again, solid content and supernatant are centrifuged after extraction again, then enzymolysis and extraction is carried out to gained solid content with homemade active liquid, again gained material and the supernatant are mixed to be put into the chromatographic column of macroreticular resin after enzymolysis and eluted, after being concentrated under reduced pressure by gained eluent and reclaim ethanol, be dried in vacuo and produce the Lilium brownie, the present invention can improve the recovery rate and purifying rate of Lilium lancifo1ium Thunb extraction saponin(e.
Description
【Technical field】
The present invention relates to field of plant extraction, especially a kind of method of Lilium lancifo1ium Thunb extraction saponin(e.
【Technical background】
Lilium lancifo1ium Thunb, it is a kind of beautiful flowers, Hua Jixiang, containing aromatic oil, spices can be made, its bulb is spherical in shape or near
Spherical, high 2.5-5 centimetres, scale white, rich in starch, the bulb of Lilium lancifo1ium Thunb is famous health food, and fresh goods can be used for
The effect of making dish, unicorn stem and flower are respectively provided with Yin nourishing and lung moistening, clearing away the heart fire and tranquillizing, available for yin chronic cough is treated, blood-stained sputum is empty
Tired palpitation with fear, people's dreaminess is removed in mistake, absent-minded to wait disease.Lily is medicine-food two-purpose product, and its nutritional ingredient mainly has protein, Duo Zhongwei
Raw element and carrotene etc., Lilium brownie contained therein have tranquilizing soporific, anti-inflammatory, antitumor, antibacterial and antiviral, reduction
The many bioactivity and pharmacological property such as cholesterol, hypoglycemic and immunological regulation, often feeding with its processed goods to human body very
It is beneficial.
In summary, lily has higher economic use value, but many ask also be present when extracting Lilium brownie
Topic, such as Lilium brownie are difficult to extract, and existing extracting method is difficult to remove all kinds of impurity therein, so as to cause lily soap
The recovery rate and purifying rate of glycosides are relatively low, it is impossible to meet the needs of current consumer groups.
【The content of the invention】
In view of posed problems above, can be more complete the invention provides a kind of method of Lilium lancifo1ium Thunb extraction saponin(e
Full holding nutritional ingredient therein, moreover it is possible to improve the recovery rate and purifying rate of Lilium lancifo1ium Thunb extraction saponin(e.
Technical scheme is as follows：
A kind of method of Lilium lancifo1ium Thunb extraction saponin(e, comprises the following steps：
(1) raw material：Full Lilium lancifo1ium Thunb bulb is chosen as raw material, after the soil for cleaning lily bulb surface, by squama
Piece separates standby from lily bulb；
(2) ultrasonication：After the scale that step (1) is isolated is cut into 5-6mm section, then scale is added thereto
The ethanol solution of 4-5 times of volume, mixed liquor is put in ultrasonic environment, and 40- is extracted under conditions of being 45-48 DEG C in temperature
Solidliquid mixture a is obtained after 45min, by the solidliquid mixture a by centrifuge in rotating speed be 10000-12000r/min
Under the conditions of centrifuge 8-10min, gained solid content a and supernatant a are retained respectively standby；
(3) microwave treatment：The ethanol solution of its 3-4 times of volume will be added in step (2) the solid content a, be in power
In 500-550W microwave abstracting equipment extract 3-4min after, be put in 50-53 DEG C of water-bath and extract 30-40min, after consolidated
Liquid mixture b, by the solidliquid mixture b by centrifuge in rotating speed be 10000-12000r/min under conditions of centrifuge 7-
9min, gained solid content b and supernatant b is retained respectively standby；
(4) enzymolysis processing：Step (3) the solid content b is put into superhigh-voltage homogenizing machine, according to 8-12g/100g use
Amount adds active liquid, handles 6-8min under conditions of pressure is 180-200MPa, temperature is 23-25 DEG C, obtains mixture c；
(5) finished product：Step (2) the supernatant a, step (3) the supernatant b and step (4) the mixture c are led to
The chromatographic column filled with macroreticular resin is crossed, after the ethanol solution for being 25-28% with volumetric concentration elutes, gained eluent is depressurized
After concentrating and reclaiming ethanol, be dried in vacuo and produce the Lilium brownie.
Further, in step (4), described active liquid is prepared by following methods：Trichoderma viride strain is forwarded to
Look into formula slant medium, 12-14h is cultivated at 20-24 DEG C, then the spore of cultured bacterial strain is put into equipped with nutrient solution
Conical flask in, vibrate 36-40min under conditions of temperature is 26-28 DEG C, hunting speed is 100-110r/min, it is then dilute
Concentration is interpreted into as 107-9Individual/mL bacteria suspension, then bacteria suspension is placed in the vibration case at uviol lamp 28-32cm,
In the lower reaction 12-18s of 180-200r/min hunting speed, active liquid is obtained.
Further, described nutrient solution includes following raw material：Ammonium sulfate, magnesium sulfate, copper chloride, iron chloride, amino acid,
Glucose, water are according to 2-3:0.01-0.02:0.02-0.04:0.02-0.03:2-3:1-2:1000 mass ratio is mixed.
Further, in step (3), the volumetric concentration of the ethanol is 70-75%.
Further, in step (2), the frequency of the ultrasonic wave is 40-50KHz.
Further, in step (2), the volumetric concentration of the ethanol is 50-55%.
The invention provides a kind of method of Lilium lancifo1ium Thunb extraction saponin(e, compared with prior art, has below beneficial to effect
Fruit：
1. of the invention first using full Lilium lancifo1ium Thunb bulb as raw material, Boli scale after being cleaned, by gained scale
It is put in ethanol solution under specified conditions, and is extracted in ultrasonic environment, solid content and upper is centrifuged after extraction
Clear liquid, then gained solid material is extracted by the way of microwave abstracting, solid content and upper is centrifuged after extraction again
Clear liquid, then enzymolysis and extraction is carried out to gained solid content with homemade active liquid, again by gained material and the supernatant after enzymolysis
Liquid mixing is put into the chromatographic column of macroreticular resin and eluted, and after being concentrated under reduced pressure by gained eluent and reclaim ethanol, carries out true
Sky is drying to obtain the Lilium brownie.Wherein, lily bud scale is put in ethanol solution under specified conditions, and in ultrasonic wave ring
Extracted in border, may separate out the compositions such as the saponins being soluble in lily bud scale in ethanol, lipid and ketone, and then after making
The efficiency of continuous extraction, and combine ultrasonic wave and extracted, ultrasonic activation that can be by particular range frequency and strong sky
Change acts on, and forces the cell rupture of lily bulb, and intracellular various polarity composition accelerates to extract in water, carried so as to shorten
The time is taken, and is advantageous to protect the activity of nutritional ingredient；Then by obtained by ultrasonic extraction material centrifuge solid content and
Supernatant, solid content therein is extracted by the way of microwave abstracting, the dipole in this method sample and solvent point
Son is in the presence of high frequency microwave energy, its positive and negative electrode of high speed velocity transformation, produces dipole vortex, ionic conduction and high-frequency and rubs
Wipe, so as to produce substantial amounts of heat in a short time, weak hydrogen bond rupture, Ion transfer etc. accelerate caused by dipole molecule rotation
Infiltration of the solvent molecule to sample matrices, i.e. raised path between farm fields destroys the histocyte wall of plant within a short period of time, forms small hole
Hole and crackle, so extracellular spe medium are just very easy to enter into the cell, dissolve and discharge intracellular material；Then
Material obtained by microwave abstracting is centrifuged into solid content and supernatant, enzyme is carried out using homemade active liquid to remaining solid content
Solution processing, wherein enzymolysis processing is carried out in superhigh-voltage homogenizing machine, and the mode that both are combined can make enzymolysis process in rapids
Carried out in the presence of the composite forces such as stream, hole, shearing, products obtained therefrom more homogeneous, emulsification, and then improve the extraction of polysaccharide can be made
Efficiency；It more can completely keep its nutritional ingredient, moreover it is possible to improve the recovery rate of Lilium lancifo1ium Thunb extraction saponin(e.The present invention is adopted
Each technological means all cooperates, mutually promoted, and camps step by step, be all linked with one another, caused total skill
Art effect be significantly larger than technological means caused by single technological means simple plus and.
2. the technique of the active liquid prepared by the present invention can efficiently prepare cellulase etc., and then be the enzymolysis of the present invention
Process provides sufficient organized enzyme, and the present invention is obtained nutrient solution using Homemade method and cultivated using Trichoderma viride as strain,
Wherein, the nutrient solution is according to specific ratio by ammonium sulfate, magnesium sulfate, copper chloride, iron chloride, amino acid, glucose, water
It is made, the requirement when disclosure satisfy that Trichoderma viride mutagenesis in the presence of nutrient solution to osmotic pressure, to promote mould mutagenesis
Energy is provided, each inorganic ions is that caused enzymatic activity provides facilitation, it is of the invention employed in technological means it is mutual
Coordinate, mutually promote, promote the preparation of active liquid from different aspect and angle, raising preparation efficiency is substantially improved.
In a word, the method for Lilium lancifo1ium Thunb extraction saponin(e provided by the invention, more can completely keep its nutritional ingredient,
The rate and purifying rate of Lilium lancifo1ium Thunb extraction saponin(e can also be improved, there is abundant nutritive value and good health-care effect.
【Embodiment】
The following examples can help those skilled in the art that the present invention is more completely understood, but cannot be with any
Mode limits the present invention.
Embodiment 1
A kind of method of Lilium lancifo1ium Thunb extraction saponin(e, comprises the following steps：
(1) raw material：Full Lilium lancifo1ium Thunb bulb is chosen as raw material, after the soil for cleaning lily bulb surface, by squama
Piece separates standby from lily bulb；
(2) ultrasonication：After the scale that step (1) is isolated is cut into 5mm section, then scale 4 is added thereto
The ethanol solution of times volume, mixed liquor is put in ultrasonic environment, is obtained after extracting 40min under conditions of being 45 DEG C in temperature
Solidliquid mixture a, by the solidliquid mixture a by centrifuge in rotating speed be 10000r/min under conditions of centrifuge 8min, point
Gained solid content a and supernatant a is not retained standby；Wherein, the volumetric concentration of the ethanol is 50%, the frequency of the ultrasonic wave
Rate is 40KHz；
(3) microwave treatment：The ethanol solution of its 3 times of volumes will be added in step (2) the solid content a, be in power
In 500W microwave abstracting equipment extract 3min after, be put in 50 DEG C of water-bath and extract 30min, after obtain solidliquid mixture b, will
The solidliquid mixture b centrifuges 7min by centrifuge under conditions of rotating speed is 10000r/min, respectively by gained solid content b
Retain with supernatant b standby；Wherein, the volumetric concentration of the ethanol is 70%；
(4) enzymolysis processing：Step (3) the solid content b is put into superhigh-voltage homogenizing machine, according to 8g/100g dosage
Active liquid is added, 6min is handled under conditions of pressure is 180MPa, temperature is 23 DEG C, obtains mixture c；
Wherein, described active liquid is prepared by following methods：Trichoderma viride strain is forwarded to and looked into formula slant medium,
12h is cultivated at 20 DEG C, then the spore of cultured bacterial strain is put into the conical flask equipped with nutrient solution, is 26 in temperature
DEG C, hunting speed vibrate 36min under conditions of being 100r/min, be then diluted to concentration as 107Individual/mL bacteria suspension, then
Bacteria suspension is placed in the vibration case at uviol lamp 28cm, in the lower reaction 12s of 180r/min hunting speed, obtained
Active liquid；
Described nutrient solution includes following raw material：Ammonium sulfate, magnesium sulfate, copper chloride, iron chloride, amino acid, glucose, water
According to 2:0.01:0.02:0.02:2:1:1000 mass ratio is mixed；
(5) finished product：Step (2) the supernatant a, step (3) the supernatant b and step (4) the mixture c are led to
The chromatographic column filled with macroreticular resin is crossed, after the ethanol solution elution that volumetric concentration is 25%, gained eluent is depressurized dense
Contract after recovery ethanol, be dried in vacuo and produce the Lilium brownie.
Embodiment 2
A kind of method of Lilium lancifo1ium Thunb extraction saponin(e, comprises the following steps：
(1) raw material：Full Lilium lancifo1ium Thunb bulb is chosen as raw material, after the soil for cleaning lily bulb surface, by squama
Piece separates standby from lily bulb；
(2) ultrasonication：After the scale that step (1) is isolated is cut into 6mm section, then scale 5 is added thereto
The ethanol solution of times volume, mixed liquor is put in ultrasonic environment, is obtained after extracting 45min under conditions of being 48 DEG C in temperature
Solidliquid mixture a, by the solidliquid mixture a by centrifuge in rotating speed be 12000r/min under conditions of centrifuge 10min, point
Gained solid content a and supernatant a is not retained standby；Wherein, the volumetric concentration of the ethanol is 55%, the frequency of the ultrasonic wave
Rate is 50KHz；
(3) microwave treatment：The ethanol solution of its 4 times of volumes will be added in step (2) the solid content a, be in power
In 550W microwave abstracting equipment extract 4min after, be put in 53 DEG C of water-bath and extract 40min, after obtain solidliquid mixture b, will
The solidliquid mixture b centrifuges 9min by centrifuge under conditions of rotating speed is 12000r/min, respectively by gained solid content b
Retain with supernatant b standby；Wherein, the volumetric concentration of the ethanol is 75%；
(4) enzymolysis processing：Step (3) the solid content b is put into superhigh-voltage homogenizing machine, according to 12g/100g dosage
Active liquid is added, 8min is handled under conditions of pressure is 200MPa, temperature is 25 DEG C, obtains mixture c；
Wherein, described active liquid is prepared by following methods：Trichoderma viride strain is forwarded to and looked into formula slant medium,
14h is cultivated at 24 DEG C, then the spore of cultured bacterial strain is put into the conical flask equipped with nutrient solution, is 28 in temperature
DEG C, hunting speed vibrate 40min under conditions of being 110r/min, be then diluted to concentration as 109Individual/mL bacteria suspension, then
Bacteria suspension is placed in the vibration case at uviol lamp 32cm, in the lower reaction 18s of 200r/min hunting speed, obtained
Active liquid；
Described nutrient solution includes following raw material：Ammonium sulfate, magnesium sulfate, copper chloride, iron chloride, amino acid, glucose, water
According to 3:0.02:0.04:0.03:3:2:1000 mass ratio is mixed；
(5) finished product：Step (2) the supernatant a, step (3) the supernatant b and step (4) the mixture c are led to
The chromatographic column filled with macroreticular resin is crossed, after the ethanol solution elution that volumetric concentration is 28%, gained eluent is depressurized dense
Contract after recovery ethanol, be dried in vacuo and produce the Lilium brownie.
Embodiment 3
A kind of method of Lilium lancifo1ium Thunb extraction saponin(e, comprises the following steps：
(1) raw material：Full Lilium lancifo1ium Thunb bulb is chosen as raw material, after the soil for cleaning lily bulb surface, by squama
Piece separates standby from lily bulb；
(2) ultrasonication：After the scale that step (1) is isolated is cut into 5.5mm section, then scale is added thereto
The ethanol solution of 4.5 times of volumes, mixed liquor is put in ultrasonic environment, after extracting 42min under conditions of being 46 DEG C in temperature
Obtain solidliquid mixture a, by the solidliquid mixture a by centrifuge in rotating speed be 11000r/min under conditions of centrifuge
9min, gained solid content a and supernatant a is retained respectively standby；Wherein, the volumetric concentration of the ethanol is 52%, described super
The frequency of sound wave is 48KHz；
(3) microwave treatment：The ethanol solution of its 3.2 times of volumes will be added in step (2) the solid content a, be in power
In 520W microwave abstracting equipment extract 3.5min after, be put in 52 DEG C of water-bath and extract 37min, after obtain solidliquid mixture b,
By the solidliquid mixture b by centrifuge in rotating speed be 11000r/min under conditions of centrifuge 8min, respectively by gained solid
Thing b and supernatant b retains standby；Wherein, the volumetric concentration of the ethanol is 72%；
(4) enzymolysis processing：Step (3) the solid content b is put into superhigh-voltage homogenizing machine, according to 9g/100g dosage
Active liquid is added, 7min is handled under conditions of pressure is 190MPa, temperature is 24 DEG C, obtains mixture c；
Wherein, described active liquid is prepared by following methods：Trichoderma viride strain is forwarded to and looked into formula slant medium,
13h is cultivated at 23 DEG C, then the spore of cultured bacterial strain is put into the conical flask equipped with nutrient solution, is 27 in temperature
DEG C, hunting speed vibrate 38min under conditions of being 105r/min, be then diluted to concentration as 108Individual/mL bacteria suspension, then
Bacteria suspension is placed in the vibration case at uviol lamp 29cm, in the lower reaction 15s of 185r/min hunting speed, obtained
Active liquid；
Described nutrient solution includes following raw material：Ammonium sulfate, magnesium sulfate, copper chloride, iron chloride, amino acid, glucose, water
According to 2.5:0.015:0.03:0.025:2.5:1.2:1000 mass ratio is mixed；
(5) finished product：Step (2) the supernatant a, step (3) the supernatant b and step (4) the mixture c are led to
The chromatographic column filled with macroreticular resin is crossed, after the ethanol solution elution that volumetric concentration is 27%, gained eluent is depressurized dense
Contract after recovery ethanol, be dried in vacuo and produce the Lilium brownie.
Comparative example 1
A kind of method of Lilium lancifo1ium Thunb extraction saponin(e, comprises the following steps：
(1) raw material：Full Lilium lancifo1ium Thunb bulb is chosen as raw material, after the soil for cleaning lily bulb surface, by squama
Piece separates standby from lily bulb；
(2) after the scale that step (1) is isolated being cut into 5.5mm section, then 4.5 times of volumes of scale are added thereto
Ethanol solution, solidliquid mixture a is obtained after soak extraction 42min under conditions of being 46 DEG C in temperature, by the solidliquid mixture a
9min is centrifuged under conditions of rotating speed is 11000r/min by centrifuge, respectively retained gained solid content a and supernatant a standby
With；Wherein, the volumetric concentration of the ethanol is 52%；
(3) microwave treatment：The ethanol solution of its 3.2 times of volumes will be added in step (2) the solid content a, be in power
In 520W microwave abstracting equipment extract 3.5min after, be put in 52 DEG C of water-bath and extract 37min, after obtain solidliquid mixture b,
By the solidliquid mixture b by centrifuge in rotating speed be 11000r/min under conditions of centrifuge 8min, respectively by gained solid
Thing b and supernatant b retains standby；Wherein, the volumetric concentration of the ethanol is 72%；
(4) enzymolysis processing：Step (3) the solid content b is put into superhigh-voltage homogenizing machine, according to 9g/100g dosage
Active liquid is added, 7min is handled under conditions of pressure is 190MPa, temperature is 24 DEG C, obtains mixture c；
Wherein, described active liquid is prepared by following methods：Trichoderma viride strain is forwarded to and looked into formula slant medium,
13h is cultivated at 23 DEG C, then the spore of cultured bacterial strain is put into the conical flask equipped with nutrient solution, is 27 in temperature
DEG C, hunting speed vibrate 38min under conditions of being 105r/min, be then diluted to concentration as 108Individual/mL bacteria suspension, then
Bacteria suspension is placed in the vibration case at uviol lamp 29cm, in the lower reaction 15s of 185r/min hunting speed, obtained
Active liquid；
Described nutrient solution includes following raw material：Ammonium sulfate, magnesium sulfate, copper chloride, iron chloride, amino acid, glucose, water
According to 2.5:0.015:0.03:0.025:2.5:1.2:1000 mass ratio is mixed；
(5) finished product：Step (2) the supernatant a, step (3) the supernatant b and step (4) the mixture c are led to
The chromatographic column filled with macroreticular resin is crossed, after the ethanol solution elution that volumetric concentration is 27%, gained eluent is depressurized dense
Contract after recovery ethanol, be dried in vacuo and produce the Lilium brownie.
Comparative example 2
A kind of method of Lilium lancifo1ium Thunb extraction saponin(e, comprises the following steps：
(1) raw material：Full Lilium lancifo1ium Thunb bulb is chosen as raw material, after the soil for cleaning lily bulb surface, by squama
Piece separates standby from lily bulb；
(2) ultrasonication：After the scale that step (1) is isolated is cut into 5.5mm section, then scale is added thereto
The ethanol solution of 4.5 times of volumes, mixed liquor is put in ultrasonic environment, after extracting 42min under conditions of being 46 DEG C in temperature
Obtain solidliquid mixture a, by the solidliquid mixture a by centrifuge in rotating speed be 11000r/min under conditions of centrifuge
9min, gained solid content a and supernatant a is retained respectively standby；Wherein, the volumetric concentration of the ethanol is 52%, described super
The frequency of sound wave is 48KHz；
(3) ethanol solution of its 3.2 times of volumes will be added in step (2) the solid content a, is put in 52 DEG C of water-bath and carries
Take 37min, after obtain solidliquid mixture b, by the solidliquid mixture b by centrifuge in rotating speed be 11000r/min condition
Lower centrifugation 8min, gained solid content b and supernatant b is retained respectively standby；Wherein, the volumetric concentration of the ethanol is 72%；
(4) enzymolysis processing：Step (3) the solid content b is put into superhigh-voltage homogenizing machine, according to 9g/100g dosage
Active liquid is added, 7min is handled under conditions of pressure is 190MPa, temperature is 24 DEG C, obtains mixture c；
Wherein, described active liquid is prepared by following methods：Trichoderma viride strain is forwarded to and looked into formula slant medium,
13h is cultivated at 23 DEG C, then the spore of cultured bacterial strain is put into the conical flask equipped with nutrient solution, is 27 in temperature
DEG C, hunting speed vibrate 38min under conditions of being 105r/min, be then diluted to concentration as 108Individual/mL bacteria suspension, then
Bacteria suspension is placed in the vibration case at uviol lamp 29cm, in the lower reaction 15s of 185r/min hunting speed, obtained
Active liquid；
Described nutrient solution includes following raw material：Ammonium sulfate, magnesium sulfate, copper chloride, iron chloride, amino acid, glucose, water
According to 2.5:0.015:0.03:0.025:2.5:1.2:1000 mass ratio is mixed；
(5) finished product：Step (2) the supernatant a, step (3) the supernatant b and step (4) the mixture c are led to
The chromatographic column filled with macroreticular resin is crossed, after the ethanol solution elution that volumetric concentration is 27%, gained eluent is depressurized dense
Contract after recovery ethanol, be dried in vacuo and produce the Lilium brownie.
Comparative example 3
A kind of method of Lilium lancifo1ium Thunb extraction saponin(e, comprises the following steps：
(1) raw material：Full Lilium lancifo1ium Thunb bulb is chosen as raw material, after the soil for cleaning lily bulb surface, by squama
Piece separates standby from lily bulb；
(2) ultrasonication：After the scale that step (1) is isolated is cut into 5.5mm section, then scale is added thereto
The ethanol solution of 4.5 times of volumes, mixed liquor is put in ultrasonic environment, after extracting 42min under conditions of being 46 DEG C in temperature
Obtain solidliquid mixture a, by the solidliquid mixture a by centrifuge in rotating speed be 11000r/min under conditions of centrifuge
9min, gained solid content a and supernatant a is retained respectively standby；Wherein, the volumetric concentration of the ethanol is 52%, described super
The frequency of sound wave is 48KHz；
(3) microwave treatment：The ethanol solution of its 3.2 times of volumes will be added in step (2) the solid content a, be in power
In 520W microwave abstracting equipment extract 3.5min after, be put in 52 DEG C of water-bath and extract 37min, after obtain solidliquid mixture b；
(4) finished product：Step (2) the supernatant a and step (3) the solidliquid mixture b are passed through filled with macropore tree
The chromatographic column of fat, after the ethanol solution elution that volumetric concentration is 27%, after being concentrated under reduced pressure by gained eluent and reclaim ethanol,
Be dried in vacuo and produce the Lilium brownie.
Comparative example 4
A kind of method of Lilium lancifo1ium Thunb extraction saponin(e, comprises the following steps：
(1) raw material：Full Lilium lancifo1ium Thunb bulb is chosen as raw material, after the soil for cleaning lily bulb surface, by squama
Piece separates standby from lily bulb；
(2) ultrasonication：After the scale that step (1) is isolated is cut into 5.5mm section, then scale is added thereto
The ethanol solution of 4.5 times of volumes, mixed liquor is put in ultrasonic environment, after extracting 42min under conditions of being 46 DEG C in temperature
Obtain solidliquid mixture a, by the solidliquid mixture a by centrifuge in rotating speed be 11000r/min under conditions of centrifuge
9min, gained solid content a and supernatant a is retained respectively standby；Wherein, the volumetric concentration of the ethanol is 52%, described super
The frequency of sound wave is 48KHz；
(3) microwave treatment：The ethanol solution of its 3.2 times of volumes will be added in step (2) the solid content a, be in power
In 520W microwave abstracting equipment extract 3.5min after, be put in 52 DEG C of water-bath and extract 37min, after obtain solidliquid mixture b,
By the solidliquid mixture b by centrifuge in rotating speed be 11000r/min under conditions of centrifuge 8min, respectively by gained solid
Thing b and supernatant b retains standby；Wherein, the volumetric concentration of the ethanol is 72%；
(4) enzymolysis processing：Step (3) the solid content b is added into active liquid according to 9g/100g dosage and handles 7min,
Obtain mixture c；
Wherein, described active liquid is prepared by following methods：Trichoderma viride strain is forwarded to and looked into formula slant medium,
13h is cultivated at 23 DEG C, then the spore of cultured bacterial strain is put into the conical flask equipped with nutrient solution, is 27 in temperature
DEG C, hunting speed vibrate 38min under conditions of being 105r/min, be then diluted to concentration as 108Individual/mL bacteria suspension, then
Bacteria suspension is placed in the vibration case at uviol lamp 29cm, in the lower reaction 15s of 185r/min hunting speed, obtained
Active liquid；
Described nutrient solution includes following raw material：Ammonium sulfate, glucose, water are according to 2.5:1.2:1000 mass ratio mixing
It is made；
(5) finished product：Step (2) the supernatant a, step (3) the supernatant b and step (4) the mixture c are led to
The chromatographic column filled with macroreticular resin is crossed, after the ethanol solution elution that volumetric concentration is 27%, gained eluent is depressurized dense
Contract after recovery ethanol, be dried in vacuo and produce the Lilium brownie.
Experimental example
Experiment is divided into 7 groups, and every group each respectively according to embodiment 1-3 and comparative example 1-4 extracting method extraction Lilium lancifo1ium Thunb
Saponin(e in bulb, the recovery rate of record gained Lilium lancifo1ium Thunb extraction saponin(e, concrete outcome are shown in Table 1.
The Lilium lancifo1ium Thunb of table 1 extracts the recovery rate of saponin(e
| Embodiment 1 | Embodiment 2 | Embodiment 3 | Comparative example 1 | Comparative example 2 | Comparative example 3 | Comparative example 4 | |
| Recovery rate (mg/g) | 3.318 | 3.265 | 3.251 | 3.011 | 2.612 | 2.137 | 2.415 |
The result of table 1 shows：The recovery rate of embodiment 1-3 Lilium lancifo1ium Thunb extraction saponin(e is higher, illustrates using the present invention
Extracting method can improve Lilium lancifo1ium Thunb saponin extraction rate.
Although above the present invention is described in detail with a general description of the specific embodiments,
On the basis of the present invention, it can be made some modifications or improvements, this will be apparent to those skilled in the art.Cause
This, these modifications or improvements, belong to the scope of protection of present invention without departing from theon the basis of the spirit of the present invention.
Claims (6)
- A kind of 1. method of Lilium lancifo1ium Thunb extraction saponin(e, it is characterised in that comprise the following steps：(1) raw material：Full Lilium lancifo1ium Thunb bulb is chosen as raw material, after the soil for cleaning lily bulb surface, by scale from Separated on lily bulb standby；(2) ultrasonication：After the scale that step (1) is isolated is cut into 5-6mm section, then scale 4-5 is added thereto The ethanol solution of times volume, mixed liquor is put in ultrasonic environment, and 40-45min is extracted under conditions of being 45-48 DEG C in temperature After obtain solidliquid mixture a, by the solidliquid mixture a by centrifuge in rotating speed be 10000-12000r/min under conditions of 8-10min is centrifuged, is respectively retained gained solid content a and supernatant a standby；(3) microwave treatment：The ethanol solution of its 3-4 times of volume will be added in step (2) the solid content a, in power be 500- After extracting 3-4min in 550W microwave abstracting equipment, be put in 50-53 DEG C of water-bath and extract 30-40min, after obtain solid-liquid and mix Compound b, by the solidliquid mixture b by centrifuge in rotating speed be 10000-12000r/min under conditions of centrifuge 7-9min, Gained solid content b and supernatant b is retained respectively standby；(4) enzymolysis processing：Step (3) the solid content b is put into superhigh-voltage homogenizing machine, added according to 8-12g/100g dosage Enter active liquid, handle 6-8min under conditions of pressure is 180-200MPa, temperature is 23-25 DEG C, obtain mixture c；(5) finished product：By step (2) the supernatant a, step (3) the supernatant b and step (4) the mixture c by filling out Chromatographic column filled with macroreticular resin, after the ethanol solution for being 25-28% with volumetric concentration elutes, gained eluent is concentrated under reduced pressure And after reclaiming ethanol, be dried in vacuo and produce the Lilium brownie.
- A kind of 2. method of Lilium lancifo1ium Thunb extraction saponin(e according to claim 1, it is characterised in that in step (4), institute The active liquid stated is prepared by following methods：Trichoderma viride strain is forwarded to and looked into formula slant medium, is cultivated at 20-24 DEG C 12-14h, then the spore of cultured bacterial strain is put into the conical flask equipped with nutrient solution, is 26-28 DEG C, vibrates in temperature Speed vibrates 30-40min under conditions of being 120-140r/min, is then diluted to concentration as 107-9Individual/mL bacteria suspension, then Bacteria suspension is placed in the vibration case at uviol lamp 28-32cm, in the lower reaction of 180-200r/min hunting speed 12-18s, obtain active liquid.
- A kind of 3. method of Lilium lancifo1ium Thunb extraction saponin(e according to claim 2, it is characterised in that described nutrient solution bag Include following raw material：Ammonium sulfate, magnesium sulfate, copper chloride, iron chloride, amino acid, glucose, water are according to 2-3:0.01-0.02: 0.02-0.04:0.02-0.03:2-3:1-2:1000 mass ratio is mixed.
- 4. a kind of method of Lilium lancifo1ium Thunb extraction saponin(e according to claim 1, it is characterised in that described in step (3) The volumetric concentration of ethanol is 70-75%.
- 5. a kind of method of Lilium lancifo1ium Thunb extraction saponin(e according to claim 1, it is characterised in that described in step (2) The frequency of ultrasonic wave is 40-50KHz.
- 6. a kind of method of Lilium lancifo1ium Thunb extraction saponin(e according to claim 1, it is characterised in that described in step (2) The volumetric concentration of ethanol is 50-55%.
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