CN101177444A - Method for extracting momordica grosvenori flavone glycoside from momordica grosvenori - Google Patents
Method for extracting momordica grosvenori flavone glycoside from momordica grosvenori Download PDFInfo
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Abstract
The invention relates to a method for extracting momordica-glycosides from fructus momordicae, which uses fresh fructus momordicae or dry fructus momordicae as material and water as extraction intermediate through multilevel counter current extraction, enzymolysis, membrane separation and column chromatography separation purification craft. The extraction yield is over 0.8% with fresh fructus momordicae as material and over 1.8% with dry fructus momordicae as material. The product purity is 60 to 90%. The product is light yellow and can be dissolved in room temperature water, which has the advantages of high product purity, light color and good solubility. Solving purification and separation problem in momordica-glycosides separation, the invention has the advantages of capability of continuous production, time and energy saving, long service life of macroporous resin and high product purity.
Description
Technical field
The present invention relates to the working method of a kind of working method of natural sweeteners, particularly Momordica-Glycosides.
Background technology
Sweet taste is one of favorite taste of people, and for a long time, sugar is the main source that the mankind obtain confectionery always, about 100,000,000 tons of year consumption of present global sugar, and year consumption is about 22 kilograms per capita.The too much person of edible sucrose easily suffers from obesity, cardiovascular, diabetes etc., and these diseases are the important factors that cause human longevity of short duration.Because these drawbacks of sugar, non-sugar sweetener arises at the historic moment, but often there are some problems in they on sense of taste, thermostability, ph stability, and simultaneously, some sintetics also has the misgivings of side effect.So their application is subjected to certain restriction.Therefore, develop the natural sugar-free functional sweetener,, have wide market application prospect to substitute high sucrose or the synthetic sweetener of heat that is using in a large number at present.
Grosvenor Momordica (Siraitia grosvenori Swingle) belongs to the Curcurbitaceae Grosvenor Momordica and belongs to, be herbaceous perennial vine place stem plant, mainly be distributed in ground such as Guangxi, Guangdong, Yunnan, Hunan, Fujian, it is one of traditional exporting of China, in Hong Kong, ground such as Macao, South East Asia and Europe have long enjoyed a good reputation, and have the title of " the refreshing fruit in east ".Grosvenor Momordica is China's distinctive medicine food dual-purpose medicinal material, and it distinguishes the flavor of sweet cool in nature, has clearing heat and moistening lung, relaxes bowel, and promotes the effect of functions of intestines and stomach.Cucurbitacin alkane three mushroom compounds (Momordica-Glycosides) are one of main effective constituent of Grosvenor Momordica, content in the Grosvenor Momordica dry fruit is about 4%, they contain common glucoside unit Fourth Ring three mushrooms (Mogrol), seven kinds have been identified at present: (1) mogroside IV (morgroside IV), (2) mogroside V (mogroside V), (3) Simon glucoside I (siamenoside I), (4) 11-oxidation-mogroside V (11-oxo-mogroside V), (5) mogroside (mogroside IIE), (6) mogroside IIIE (mogroside IIIE), (7) mogroside III (mogrosideIII).The sweet glucoside of this class in the Grosvenor Momordica is sweet taste mostly, and wherein beet aglucone V (Morgroside V) is main sweet ingredient, and Momordica-Glycosides content was compared with sucrose greater than 80% o'clock, and sugariness is greater than 200 times of sucrose, and heat has only 1/50th of sucrose.Neutral aqueous solution or in 120 ℃ of air long-time heating, Momordica-Glycosides is not destroyed, Heat stability is good.Momordica-Glycosides is stable under the sour environment condition, studies show that to preserve in pH value 2.0-10.0 scope any variation can not take place in 2 years.Simultaneously, deposit separately or with other material close put can not take place in 18 months the fermentation or mildew phenomena.Acute toxicity test shows that product is nontoxic level.In addition, Momordica-Glycosides also has health-care effect, and pharmacology identifies and to show, effects such as Momordica-Glycosides has antibechic, eliminates the phlegm, eases pain, relaxes bowel, regulates digestion, protects the liver step-down, enhancing immunity and antibacterial growth.Therefore, Momordica-Glycosides be that a kind of sugariness height, heat are low, the functional sweetener natural low in calories of lighter color, good water solubility, good stability.
Extract Momordica-Glycosides at present from Grosvenor Momordica, the method for use adopts a jar group extraction method to extract mostly, is solvent with water or ethanol, through centrifugal removal of impurities, handle by decolorizing resin or the decolouring of Al2O3 post, macroporous resin is refining, more dried the forming of spray behind concentrate drying.As the patent No. is that the Chinese patent of ZL87101850 discloses " a kind of extracting method of Momordica-Glycosides ", and it is to grind, beat oar with the Grosvenor Momordica fresh fruit, and warm water 50-60 degree extracts, filtration.Separate by DA-201 acrylic type sorbent material, 50% (V/V) ethanol elution, through the decolouring of D-211 acrylic type resin anion(R.A), desalination, decon, distillation method reclaims ethanol, and through the vacuum pump concentrating under reduced pressure, lyophilize obtains Momordica-Glycosides.And for example number of patent application is that the patent application document of ZL200510055229.6 discloses a kind of method of extracting Momordica-Glycosides from Grosvenor Momordica, it be a kind of be raw material with the Grosvenor Momordica, with water or low-carbon alcohol is solvent, extract through solvent, concentrate, technologies such as separation and purification, extracting method is water heating extraction or microwave extraction or ultrasonic extraction, separation and purification is column chromatographic isolation and purification or membrane separation purification, the sweet salidroside content that this method is extracted is 40-80% by the ultraviolet spectrometry spectrographic determination, or is 20-40% by high effective liquid chromatography for measuring Momordica grosvenori mogroside V weight content.This patented method has following shortcoming: 1) adopt single jar or pot group type extraction, water consumption is big and the time is long, and extraction efficiency is not high; Extraction temperature height, heat energy loss is big; 2) contain a large amount of pectin in the Grosvenor Momordica fresh fruit, extracting solution through micro-filtration and centrifugal after use macroporous resin adsorption, pectin can't effectively be removed, the resin vulnerable to pollution, resin work-ing life is short; 3) then use macroporous resin adsorption by decolouring, the efficient of decolorizing column is not high, and the color and luster of product is darker, and product purity is not high, only is 80%.4) by spraying drying, the product solvability is relatively poor relatively, needs can use with the hot water dissolving, and mouthfeel is not good enough yet simultaneously.Therefore, above-mentioned two kinds of processing methodes can't serialization production, and production efficiency is low, and of low quality in the product that makes, extracting method haves much room for improvement.
Summary of the invention
The invention provides a kind of method of extracting Momordica-Glycosides from Grosvenor Momordica, have and can be continuously produced, the production efficiency height extracts yield and product purity height, the characteristics of the long service life of resin.
The present invention extracts Momordica-Glycosides from Grosvenor Momordica, be raw material with Grosvenor Momordica fresh fruit or dry fruit, with water is to extract medium, comprises following processing step:
1) extraction process: Grosvenor Momordica is pulverized the back and is adopted the multi-stage countercurrent extraction method to carry out, and lixiviate 60-90 minute, extracting temperature was 50-90 ℃, makes extracting solution after coarse filtration.The preferred three stage countercurrent extraction methods that adopt, extracting temperature is 60-70 ℃.Grosvenor Momordica fresh fruit or dry fruit should be ground into the smalls shape, and granularity is not more than 20 orders. and when being raw material with the Grosvenor Momordica fresh fruit, the water extraction medium is 10-15 a times of fresh fruit weight; When being raw material with the Grosvenor Momordica dry fruit, the water extraction medium is 20-30 a times of fresh fruit weight.The advantage of employing countercurrent extraction method is the move toward one another by current and material, make water fully contact with material, water and material have remained effective concentration difference between difference extraction level, the pure water of one-level extracts is material slag after secondary extracts, and what the secondary extracting solution extracted is the material slag of three grades of extractions.Thereby make material can access abundant extraction, used extraction time shortens greatly, has both guaranteed the extraction yield, has shortened extraction time, also saves the consumption of water.Simultaneously, utilize Momordica-Glycosides characteristic soluble in water, adopt extract at low temperature, reduced thermal losses.
2) membrane separation process: with the extracting solution via hole diameter is after the stainless steel microfiltration membrane of 0.3 μ m-0.5 μ m is filtered, is cooled to 18-22 ℃ and carries out the centrifugation clarification, and be that 40,000-100,000 daltonian ultra-filtration membranes carry out ultrafiltration with holding back relative molecular weight then.Extracting solution is cooled to 18-22 ℃ can make macromolecular substance wherein form floss, because these materials can't be removed by centrifuging, must could clarify by ultrafiltration.The stainless steel microfiltration membrane can be removed fine particle, centrifugation part that can be removed pectin, protein and other, and partial pigment or other impurity can be further removed in ultrafiltration.Behind membrane sepn, avoided occurring floss in the extracting solution, guaranteed the clarity of extracting solution, thereby can farthest reduce pollution polymeric adsorbent.Be preferably in the above-mentioned steps: stainless steel microfiltration membrane aperture is 0.5 μ m, the extracting solution temperature is cooled to 20 ℃, separate with the butterfly centrifugal machine, centrifuge speed is 6000 rev/mins again, and ultra-filtration membrane is selected for use and held back relative molecular weight is 60,000 daltonian hollow fiber ultrafiltration membrane.
3) concentration technology: adopt vacuum concentration or reverse osmosis membrane concentration method that extracting solution is concentrated into 5-10 hundred sharp degree (Brix).When wherein adopting vacuum concentration, temperature is 50-70 ℃, pressure 0.08-0.12Mpa; Reverse osmosis membrane adopts and holds back relative molecular weight is 200 daltonian RO films, operating pressure 2.0-2.5Mpa.
4) separation purifying technique: the extracting solution after will concentrating adsorbs with macroporous adsorbent resin, when feed liquid reaches leakage point, cleans with deionized water, to remove remaining impurities and partial pigment; Carry out wash-out with 50% ethanol again, do not have sweet taste, collect ethanol eluate, again ethanol eluate is reclaimed ethanol by the ethanol recovery tower until effluent liquid.Aforesaid macroporous adsorbent resin can be selected D101 or AB-8 for use.
5) drying process: adopt vacuum concentration will be concentrated into through the extracting solution after the separation and purification more than the 1500 sharp degree, carry out lyophilize again.
When being the raw material extraction with the Grosvenor Momordica fresh fruit, also comprise enzymolysis process, promptly in the extracting solution that extraction process makes, add 3/10000ths to 10/10000ths pectin compound enzyme and carry out enzymolysis, the pH value is 3.5-6.0, hydrolysis temperature is 30-60 ℃, pectin compound enzyme is the mixture that contains polygalacturonase and hemicellulase, and wherein the ratio of polygalacturonase and hemicellulase is 2: 1-3: 1.The ratio that is preferably polygalacturonase and hemicellulase is 3: 1, and the pH value is 4.5-5.0, and hydrolysis temperature is 40-45 ℃.Owing to contain a large amount of pectin in the Grosvenor Momordica fresh fruit, if can not effectively remove, very easily cause the pollution of resin in the subsequent handling, therefore, Grosvenor Momordica fresh fruit extracting solution must be removed the pectin in the feed liquid after coarse filtration.After enzymolysis is complete, carry out the processing of subsequent handlings such as centrifugal and ultrafiltration again.
Be to reduce pigment content in the finished product, improve product purity, in aforesaid separation purifying technique, can be earlier with ethanol eluate by anionite-exchange resin decolour handle after, again ethanol eluate is reclaimed ethanol by the ethanol recovery tower.Wherein preferred anionic surfactants exchange resin model is D-280.
The inventive method is a raw material with Grosvenor Momordica fresh fruit or dry fruit, with water is to extract medium, uses multi-stage countercurrent extraction, enzymolysis, membrane separation technique and column chromatographic isolation and purification technology, has solved removal of impurities, separation, thickening problem during Momordica-Glycosides separates.Have serialization production, time-saving energy-saving, the macroporous resin life-span is long, product purity is high characteristics.According to the inventive method, be raw material with the fresh fruit, to extract yield and can reach more than 0.8%, the yield of dry fruit can reach more than 1.8%, product purity 60-90%; The little Huang of the product color of producing is a solubilized with warm water, has product purity height, lighter color, characteristics that solvability is good.
Embodiment
Example one:
1, fruit is handled: the Grosvenor Momordica fresh fruit of gathering, and In Shade spreading for cooling 3-7 days, make in the fruit and may fully transform by remaining little bitter substance, reject decayed fruit, mould fruit, to pulverize with pulverizer or hollander, fineness is less than 20 orders.
2, three stage countercurrents extract: 1000 kilograms of ratios with water of the fresh fruit after the pulverizing fed intake by 1: 15, divided the 20 minutes equivalent in three every intervals of period to drop in the counterflow extraction apparatus, extracted 60 ℃ of temperature, extracted 75 minutes.
3, enzymolysis processing: extracting solution through coarse filtration, be cooled to 40-50 ℃, added 5/10000ths pectin compound enzyme enzymolysis 60 minutes.
4, the extracting solution behind the enzymolysis is cooled to 20 degree rapidly through 0.5 μ m micro-filtrate membrane filtration, carries out centrifugation with the butterfly centrifugal machine with 6000 rev/mins then, is that 60,000 daltonian hollow cellulose films carry out ultrafiltration through the relative molecular weight that dams again.
5, with the extracting solution after the ultrafiltration through vacuum concentrator, be 0.06-0.1 at 50-55 ℃, vacuum tightness, be concentrated into 600 sharp degree (Brix).
6, the extracting solution after will concentrating with certain flow velocity by macroporous adsorptive resins until the leak point, discard effluent liquid, then with the washed with de-ionized water of 1 column volume.
7, carry out wash-out till effluent liquid does not have sweet taste with 50% ethanolic soln, collect ethanol eluate.
8, ethanol eluate reclaims ethanol through the ethanol recovery tower, will remove the alcoholic acid extracting solution again and carry out vacuum concentration, and to 1,500 sharp degree (Brix), finished product is made in lyophilize then, the sterilization packing.
Make 11 kilograms of Momordica-Glycosides, extracting yield is 1.1%, and the Momordica-Glycosides weight content is more than 60% in the extract, and the content of Momordica grosvenori mogroside V is more than 20%, and color and luster is yellowish, and solvability is good, is solubilized with normal-temperature water.
Example two:
1, fruit is handled: the Grosvenor Momordica fresh fruit of gathering, and In Shade spreading for cooling 3-7 days, make in the fruit and may fully transform by remaining little bitter substance, reject decayed fruit, mould fruit, to pulverize with pulverizer or hollander, fineness is less than 20 orders.
2, three stage countercurrents extract: 1000 kilograms of ratios with water of the fresh fruit after the pulverizing fed intake by 1: 15, divided the 20 minutes equivalent in three every intervals of period to drop in the counterflow extraction apparatus, extracted 60 ℃ of temperature, extracted 75 minutes.
3, enzymolysis processing: extracting solution through coarse filtration, be cooled to the 40-50 degree, added 5/10000ths pectin compound enzyme enzymolysis 60 minutes.
4, the extracting solution behind the enzymolysis is cooled to 20 ℃ rapidly through 0.5 μ m micro-filtrate membrane filtration, carries out centrifugation with the butterfly centrifugal machine under 6000 rev/mins then, is that 60,000 daltonian hollow cellulose films carry out ultrafiltration through the relative molecular weight that dams again.
5, with the extracting solution after the ultrafiltration through vacuum concentrator, thickening temperature is that 50-55 ℃, vacuum tightness are 0.06-0.1, is concentrated into 600 sharp degree (Brix).
6, the extracting solution after will concentrating with certain flow velocity by macroporous adsorptive resins until the leak point, discard effluent liquid, then with the washed with de-ionized water of 1 column volume.
7, carry out wash-out till effluent liquid does not have sweet taste with 50% ethanolic soln, collect ethanol eluate.
8, the ethanol eluate elutriant is decoloured through anionite-exchange resin with certain flow velocity, collect destainer.
9, destainer reclaims ethanol through the ethanol recovery tower, and more than vacuum concentration to the 15 hundred sharp degree (Brix), finished product is made in lyophilize then, the sterilization packing.
Make Momordica-Glycosides and be about 8 kilograms, extracting yield is 0.8%, and the Momordica-Glycosides weight content is more than 85% in the extract, and the content of Momordica grosvenori mogroside V is more than 35%, the little Huang of color and luster, and solvability is good, is solubilized with normal-temperature water.
Example three:
1, fruit is handled: with pulverizer 600kg Grosvenor Momordica dry fruit is pulverized, fineness is less than 20 orders.
2, three stage countercurrents extract: the fruit after the pulverizing and the ratio of water fed intake by 1: 25, according to the amount of the water in the extraction equipment, divided three period equivalent to drop in the counterflow extraction apparatus, extracted 80 ℃ of temperature, 75 minutes extraction times.
3, with extracting solution through 0.5 μ m micro-filtrate membrane filtration, be concentrated into 600 sharp degree (Brix) with vacuum concentrator again, thickening temperature is that 50-55 ℃, vacuum tightness are 0.06-0.1.Being cooled to 20 ℃ then rapidly, carrying out centrifugation with the butterfly centrifugal machine under 6000 rev/mins, to remove macromole impurity, is that 40,000 daltonian hollow cellulose films carry out ultrafiltration through the relative molecular weight that dams again.
4, extracting solution that will be after ultrafiltration is through vacuum concentrator, and thickening temperature is that 50-55 ℃, vacuum tightness are 0.06-0.1, is concentrated into 4-6 hundred sharp degree (Brix).
5, the extracting solution after will concentrating with certain flow velocity by macroporous adsorptive resins until the leak point, discard effluent liquid, then with the washed with de-ionized water of 1 column volume.
6,, carry out wash-out till effluent liquid does not have sweet taste with 50% ethanolic soln, the collection ethanol eluate.
7, the ethanol eluate elutriant is decoloured through anionite-exchange resin with certain flow velocity, collect destainer.
8, destainer reclaims ethanol through the ethanol recovery tower, and more than vacuum concentration to the 15 hundred sharp degree (Brix), finished product is made in lyophilize then, the sterilization packing.
Make 10 kilograms of Momordica-Glycosides, extracting yield is 1.7%, and the Momordica-Glycosides weight content is more than 90% in the extract, and the content of Momordica grosvenori mogroside V is more than 40%, the little Huang of color and luster, and solvability is good, is solubilized with normal-temperature water.
Above-mentioned is the specific embodiment of the present invention, but design concept of the present invention is not limited thereto, and allly utilizes this design that the present invention is carried out the change of unsubstantiality, all belongs to the behavior of invading protection domain of the present invention.
Claims (10)
1. a method of extracting Momordica-Glycosides from Grosvenor Momordica is a raw material with Grosvenor Momordica fresh fruit or dry fruit, is to extract medium with water, it is characterized in that comprising following processing step:
1) extraction process: adopt the multi-stage countercurrent extraction method to carry out lixiviate in Grosvenor Momordica fresh fruit or dry fruit pulverizing back, extracting temperature is 50-90 ℃, makes extracting solution after coarse filtration;
2) membrane separation process: with the extracting solution via hole diameter is after the stainless steel microfiltration membrane of 0.3 μ m-0.5 μ m is filtered, to be cooled to 18-22 ℃ and to carry out centrifugation, is that 40,000-100,000 daltonian ultra-filtration membranes carry out ultrafiltration with holding back relative molecular weight then;
3) concentration technology: adopt vacuum concentration or reverse osmosis membrane concentration method that extracting solution is concentrated into 5-10 hundred sharp degree;
4) separation purifying technique: the absorption with macroporous adsorbent resin Momordica-Glycosides of the extracting solution after will concentrating, use the washed with de-ionized water macroporous adsorbent resin then, carry out wash-out with 50% ethanol again, again ethanol eluate is reclaimed ethanol by the ethanol recovery tower;
5) drying process: adopt vacuum concentration will be concentrated into through the extracting solution after the separation and purification more than the 1500 sharp degree, carry out lyophilize again.
2. according to the described a kind of method of from Grosvenor Momordica, extracting Momordica-Glycosides of claim 1, it is characterized in that: when being raw material with the Grosvenor Momordica fresh fruit, also comprise enzymolysis process, promptly in the extracting solution that extraction process makes, add 3/10000ths to 10/10000ths pectin compound enzyme and carry out enzymolysis, the pH value is 3.5-6.0, and hydrolysis temperature is 30-60 ℃.
3. according to the described a kind of method of from the Grosvenor Momordica fresh fruit, extracting Momordica-Glycosides of claim 2, it is characterized in that: described pectin compound enzyme is the mixture that contains polygalacturonase and hemicellulase, and wherein the ratio of polygalacturonase and hemicellulase is 2: 1-3: 1.
4. according to the described a kind of method of extracting Momordica-Glycosides from Grosvenor Momordica of claim 1, it is characterized in that: described multi-stage countercurrent extraction method is that three stage countercurrents extract, and extracting temperature is 60-70 ℃.
5. according to the described a kind of method of extracting Momordica-Glycosides from Grosvenor Momordica of claim 1, it is characterized in that: when described raw material was the Grosvenor Momordica fresh fruit, the extraction medium was 10-15 a times of fresh fruit weight; When described raw material was the Grosvenor Momordica dry fruit, the water extraction medium was 20-30 a times of fresh fruit weight.
6. according to the described a kind of method of extracting Momordica-Glycosides from Grosvenor Momordica of claim 1, it is characterized in that: described centrifugation adopts the butterfly centrifugal machine to separate.
7. according to the described a kind of method of extracting Momordica-Glycosides from Grosvenor Momordica of claim 1, it is characterized in that: described ultra-filtration membrane is 4-10 ten thousand daltonian hollow fiber ultrafiltration membrane for holding back relative molecular weight.
8. according to the described a kind of method of extracting Momordica-Glycosides from Grosvenor Momordica of claim 1, it is characterized in that: described vacuum concentration temperature is 50-70 ℃, and concentrated pressure is 0.08-0.12Mpa.
9. according to the described a kind of method of extracting Momordica-Glycosides from Grosvenor Momordica of claim 1, it is characterized in that: described reverse osmosis membrane is 200 daltonian RO films for holding back relative molecular weight, operating pressure 2.0-2.5Mpa.
10. according to the described a kind of method of from Grosvenor Momordica, extracting Momordica-Glycosides of claim 1, it is characterized in that: in the described separation purifying technique, ethanol eluate after anionite-exchange resin decolours processing, reclaims ethanol with ethanol eluate by the ethanol recovery tower earlier again.
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2006
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