CN104098713B - A kind of method simultaneously preparing garlic polysaccharide and alliin - Google Patents

Abstract

The present invention discloses a kind of method simultaneously preparing garlic polysaccharide and alliin, relates to a kind of applied microwave inactivation allinase, membrance separation, ion exchange and crystallization technique.Its specifically comprises the processes of: the allinase in applied microwave drying machinery inactivation fresh garlic, application soy bean milk making machine making beating, water extraction, filter, tubular membrane is clarified, and the membranous system of molecular cut off 3KD filters；Molecular weight is more than 3KD part, the most i.e. obtains garlic polysaccharide product；Molecular weight is less than 3KD part, adjusts pH=2, crosses cationic exchange resin adsorption and concentrates, and water washs, ammonia eluting, collects ammonia elution fraction, concentrates, and application alcohol crystal, recrystallization i.e. obtain alliin product.The present invention is prepared for garlic polysaccharide and alliin simultaneously, adds utilizability and the economic benefit of Bulbus Allii；Before ion exchange, not concentrating, direct upper prop adsorbs, and has not only saved the energy, and has decreased the loss of alliin, has improve yield.

Description

A kind of method simultaneously preparing garlic polysaccharide and alliin

Technical field

The present invention relates to phytochemistry, food field of deep, specifically a kind of applied microwave, membrance separation, from Son exchange and crystallization technique, combine the method preparing garlic polysaccharide with alliin from fresh garlic.

Background technology

Alliin (SACS, alliin) is the abundantest organic of content inside Bulbus Allii Sulfide, accounts in Bulbus Allii the 90% of sulfur-containing compound.Recent study shows that alliin has antioxidation, removing Free radical, protect the liver, blood sugar lowering, blood fat reducing, trophic nerve, antitumor, antithrombotic, the multiple function such as antibacterial (merchant Jiang Bin, etc. the progress of sulfur-containing amino acid in Bulbus Allii. Chinese herbal medicine, 2000,31 (6): 468-490；Wang Minglu, greatly The extraction and separation process research of alliin in Bulbus Allii. Southern Yangtze University, 2012), in Medicines and Health Product, food additive With the prospect that the fields such as cosmetics have wide application.

Have 10 patents of invention at present from Bulbus Allii, prepare the extract containing alliin or separate alliin.In State's patent 00101244.4 is with freeze-dried garlic powder as raw material, and application ethanol extraction, concentration, silica gel column chromatography are contained The solid content of alliin.Enzyme denaturing, pectase, cellulase degradation are boiled in Chinese patent 02152304.5 application Removing garlic polysaccharide with fermentable, then the step such as cation exch ange adsorption, crystallization extracts Bulbus Allii from fresh Bulbus Allii Propylhomoserin.Patent 03100420.2 uses microwave deactivating enzyme, ethanol extraction, concentration, cation exch ange adsorption, knot Crystalline substance prepares alliin.Chinese patent 200310117625.8 invention application cation switched segment collects associating The method preparing garlic polysaccharide and alliin.Chinese patent 200610027751.8 uses freezing enzyme denaturing, ethanol Extraction, copper acetate precipitation, ethanol precipitation, activated carbon decolorizing, concentrate, the step such as crystallization prepares alliin.In State's patent 200910242303.3 applies tunnel type micro wave technology to inactivate allinase, applies multienzymatic reaction system Protein in the middle of degraded Bulbus Allii, pectin, the thick substances such as cellulose, in the fresh Bulbus Allii of ion exchange extraction purification Alliin.Chinese patent 201110139510.3 applies enzyme denaturing, water extraction, tune pH to be centrifuged, ion-exchange absorption Attached, nanofiltration, drying and other steps prepare alliin.Chinese patent 201010267757.9 applies polarity organic molten Bulbus Allii, ultrasonication, ultrafiltration, nanofiltration, condensing crystallizing are soaked in agent, prepare alliin.China 201210322853.8 Application heating enzyme denaturing, making beating, supersound extraction, ultrafiltration Deproteinization polysaccharide, lyophilization, prepare alliin.In State's patent 200910142157.7 applies ethanol or acetone recrystallization to prepare alliin reference substance.

Bulbus Allii contains 15%～25% sugar, predominantly polysaccharide, on the one hand garlic polysaccharide severe jamming alliin Purification；On the other hand, garlic polysaccharide has the guarantors such as antioxidation, antiviral, enhancing immunity, the liver protecting Health-care function (Diao Juanjuan, the pharmacognosy of Bulbus Allii and garlic polysaccharide quality research. Xinjiang Medicine University, 2006), tool Have extraction to separate to be worth.Chinese patent 98113263.4 applies steaming and decocting, centrifugal, concentrations, ultrafiltration, dry etc. Method extracts garlic polysaccharide from Bulbus Allii, but have lost alliin.

Should except Chinese patent 200310117625.8 (CN1556102, publication date December in 2004 22 days) Combine with cation exchange resin and prepare outside garlic polysaccharide and alliin, prepared the patent of alliin except existing Power consumption high (freezing enzyme denaturing, secondary concentration), can not industrialization or commercial viability the highest (organic solvent carries Take) etc. outside problem, all extract the garlic polysaccharide being worth abandon having, waste resource.And Chinese patent 200310117625.8 " combine the method preparing alliin with garlic polysaccharide " and use ion-exchange chromatography and divide Section eluting prepares the technique of garlic polysaccharide and alliin simultaneously, and its commercial viability is the highest.

Summary of the invention

The technical problem to be solved in the present invention is to overcome to waste garlic polysaccharide in prior art or industrialization is feasible Spend the most high defect.The technology such as a kind of application film, ion exchange, crystallization that it is an object of the invention to provide are made simultaneously Standby garlic polysaccharide and the method for alliin.First the present invention applies membrane technology to prepare garlic polysaccharide, filters liquid direct Ion-exchange absorption is attached, crystallize, recrystallization prepares alliin, and the present invention proposes new technical scheme, with existing Technology is compared, and the invention have the advantages that (1) technique is simple, easy and simple to handle.It is prepared for Bulbus Allii simultaneously Polysaccharide, adds utilizability and the economic benefit of Bulbus Allii；(2) before ion-exchange chromatography, film is applied Technology has separated the macromolecular substances such as garlic polysaccharide, significantly reduces the impurity affecting alliin absorption, improves Chromatography efficiency, decreases the loss of resin；(3) this method is before ion exchanges, and does not concentrates, directly on Post adsorbs, and has not only saved the energy, and has decreased the loss of alliin, has improve yield.

A kind of method simultaneously preparing garlic polysaccharide and alliin provided by the present invention, its step is as follows:

(1), after fresh garlic tap water washes impurity, allinase, spreading for cooling are inactivated with microwave tea-steaming machine After, add distilled water, with soy bean milk making machine pull an oar, and filter removal Bulbus Allii slag obtain Bulbus Allii slurry, the addition of distilled water is 15 times of Bulbus Allii weight；

(2) filtering Bulbus Allii slurry with gauze or absorbent cotton, filtrate applies tubular membrane to clarify to obtain clarification Bulbus Allii slurry further, uses Molecular cut off is that 3KD membrane filtration system filters clarification Bulbus Allii slurry；

(3) by molecular weight more than 3KD part, concentrate and be dried, obtain garlic polysaccharide extract；

(4) by molecular weight less than 3KD part, regulate pH=2, swapped by highly acidic cation exchange column With concentration, successively with distilled water, 0.5M ammonia eluting, collect ammonia elution fraction；Distillation used by eluting The consumption of water is 6 times of highly acidic cation exchange column volume, and the consumption of the 0.5M ammonia used by eluting is strong 4 times of acidic cation exchange column volume；

(5) in 50 DEG C of rotary evaporations, the ammonia elution fraction of collection being evaporated to alliin concentration is The liquid of 180mg/ml～220mg/ml, adds dehydrated alcohol, and the addition of dehydrated alcohol is described alliin Concentration is 2～3 times of the liquid volume of 180mg/ml～220mg/ml, and placement crystallizes and i.e. obtains white needles knot Brilliant thing；

(6) after white, needle-shaped crystals thing being filtered, more successively with the dehydrated alcohol that volume fraction is 80%, volume integral Number be 90% ethyl alcohol recrystallization i.e. obtain purity more than 98% alliin crystallization.

Accompanying drawing explanation

Fig. 1 is the HPLC-UV detection using efficient liquid phase liquid spectrometry to measure alliin crystallization purity.Horizontal seat in figure Mark is retention time, and vertical coordinate is ultraviolet absorption value.

Detailed description of the invention

Without illustrating as conventional method in following embodiment.

Embodiment 1

(1) fresh garlic 100kg, tap water washs, and after removing the impurity such as soil, heats 7 with microwave tea-steaming machine Minute, inactivate allinase；After spreading for cooling, add the distilled water of 1500kg, pull an oar with commercial soymilk grinder, and Filter removal Bulbus Allii slag and obtain Bulbus Allii slurry.

(2) starching with gauze or absorbent cotton Filtration Filtration Bulbus Allii, filtrate applies tubular membrane to clarify to obtain clarification Bulbus Allii slurry further, Again with molecular cut off be 3KD membrane filtration system filter clarification Bulbus Allii slurry.

(3) molecular weight is more than the solvent portions of 3KD, concentrates and be dried, obtain garlic polysaccharide extract.

(4) molecular weight is less than the solvent portions of 3KD, hydrochloric acid regulation pH=2, is exchanged by highly acidic cation Post swaps and concentrates, and successively with distilled water and 0.5M ammonia eluting, collects ammonia elution fraction；Eluting 6 times that consumption is highly acidic cation exchange column volume of distilled water used, the 0.5M ammonia used by eluting 4 times that consumption is highly acidic cation exchange column volume.

(5) in 50 DEG C of rotary evaporations, the ammonia elution fraction of collection being evaporated to alliin concentration is 200mg/ml Liquid, add dehydrated alcohol, the addition of dehydrated alcohol be described alliin concentration be the liquid of 200mg/ml 3 times of body volume, placement crystallizes and i.e. obtains white, needle-shaped crystals thing.

(6) after white, needle-shaped crystals thing being filtered, more successively with the dehydrated alcohol that volume fraction is 80% and volume integral Number be 90% ethyl alcohol recrystallization i.e. obtain 500g purity more than 98% alliin crystallization.

Embodiment 2

(1) content using the polysaccharide of sulphuric acid anthrone method mensuration garlic polysaccharide extract is 20-30%.

(2) efficient liquid phase liquid spectrometry is used to measure alliin crystallization purity > 99% (Fig. 1).

Claims (1)

1. the method simultaneously preparing garlic polysaccharide and alliin, it is characterised in that:

(1), after fresh garlic tap water washes impurity, allinase is inactivated with microwave tea-steaming machine, After spreading for cooling, add distilled water, with soy bean milk making machine pull an oar, and filter removal Bulbus Allii slag obtain Bulbus Allii slurry, the addition of distilled water Amount is 15 times of Bulbus Allii weight；

(2) filtering Bulbus Allii slurry with gauze or absorbent cotton, filtrate applies tubular membrane to clarify to obtain clarification Bulbus Allii slurry further, It is that 3KD membrane filtration system filters clarification Bulbus Allii slurry with molecular cut off；

(3) molecular weight is more than the solvent portions of 3KD, concentrates and be dried, obtain garlic polysaccharide extract；

(4) molecular weight is less than the solvent portions of 3KD, regulates pH=2, exchanged by highly acidic cation Post swaps and concentrates, and successively with distilled water, 0.5M ammonia eluting, collects ammonia elution fraction；Eluting 6 times that consumption is highly acidic cation exchange column volume of distilled water used, the 0.5M ammonia used by eluting 4 times that consumption is highly acidic cation exchange column volume；

(5) in 50 DEG C of rotary evaporations, the ammonia elution fraction of collection being evaporated to alliin concentration is The liquid of 180mg/mL～220mg/mL, adds dehydrated alcohol, and the addition of dehydrated alcohol is described alliin Concentration is 2～3 times of the liquid volume of 180mg/mL～220mg/mL, and placement crystallizes and i.e. obtains white needles knot Brilliant thing；

(6) after white, needle-shaped crystals thing being filtered, more successively with the dehydrated alcohol that volume fraction is 80%, body Fraction be 90% ethyl alcohol recrystallization i.e. obtain purity more than 98% alliin crystallization.