CN1575663A - Sea cucumber mucopolysaccharide-riched food and making method thereof - Google Patents
Sea cucumber mucopolysaccharide-riched food and making method thereof Download PDFInfo
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Abstract
Sea cucumber as material is produced into pure mucoitin product through alkaline hydrolysis, enzymolysis with single or double foreign enzyme or the combination of autoenzyme and foreign enzyme; the alcoholysis of the separated mother liquid; and purification. The pure mucoitin product may be added into spirit to obtain sea cucumber wine; the pure mucoitin product may be mixed with Chinese medicine extract liquid and the mixture is further processed to obtain granule, oral liquid and tablet; and the pure mucoitin product may be mixed with beta-cyclodextrin and the mixture is boiled, vacuum dried and capsulized to obtain capsule. The present invention extracts mucoitin from sea cucumber and produces various functional foods with certain mucoitin content with the functions of resisting cancer and raising immunity.
Description
The application divides an application August 30 2002 original application day, original applying number 02132846.3 for " being rich in sea cucumber mucopolysaccharide food and preparation method thereof ".
Technical field the present invention relates to a kind of food, particularly marine product food.
Background technology is well-known, and sea cucumber mucopolysaccharide has the immunity of enhancing, antitumor action.But the cucumber product of Xiao Shouing mostly is full ginseng product in the market, though it contains sea cucumber mucopolysaccharide, its content seldom is difficult to its due effect of performance.
Summary of the invention the present invention is to be raw material with the sea cucumber, adopts modern biotechnology and food new and high technology to extract sea cucumber mucopolysaccharide, and is made into the method for multiple functional health care food.
One, the extraction of sea cucumber mucopolysaccharide
1, raw material and processing: get dried sea-cucumber and foam; Or get the salt marsh sea cucumber, and vertically cut open along a side jaggy with cutter, remove foreign material in ginseng mouth and the chamber, in clear water, clean; Or get fresh sea cucumber, directly remove internal organ, remove muscle, clean.The water that adds 0～5 times of volume then to above-mentioned sea cucumber, with tissue mashing machine with its wall fragmentation.
2, extracting method
1. alkaline hydrolysis combines with single enzymolysis
Under 40～70 ℃ of conditions, add 0.2～0.8% solid K
2CO
3, stir hydrolysis down 1～5 hour.Transfer pH7～9 with 6 mol hydrochloric acid afterwards, (and keeping the pH value in this scope in enzymolysis process), the adding mass fraction is 0.3～0.5% trypsase (enzyme alive 5 * 10
4U/g) maintain the temperature at 45～55 ℃, enzymolysis is 2～7 hours under stirring.Speed is chilled to below 30 ℃ afterwards, keeps cessation reaction after 30 minutes.Transfer pH to neutral with 6 mol hydrochloric acid, high speed centrifugation (6000 rev/mins) separated 15 minutes, the sediment that obtains is insoluble hydrolysate, wherein contain the albumen about 12%, can be as base-material, it further is processed into powdery, graininess, tablet or makes series of products such as oral liquid through specially treated.To centrifugal mother liquid obtained 95% ethanol (mass percent is 30～80%) that slowly adds, constantly stir this moment, and in 10 ℃ of following temperature overnight.Centrifugal afterwards, the gained supernatant can carry out decompression distillation, not only recyclable ethanol, can also obtain the sea cucumber hydrolysate, can it further be processed into various functional food, collect centrifugal sediment for base-material, and successively with the washing of 90～100% ethanol, acetone dehydration, in 40～70 ℃ of dryings or vacuum drying, the product that obtains is the sea cucumber mucopolysaccharide crude product.
2. alkaline hydrolysis combines with double-enzyme hydrolysis
Under 40～70 ℃ of conditions, add 0.2～0.8% solid K
2CO
3, stir hydrolysis down 1～5 hour, transfer pH1～5 with 6 mol hydrochloric acid afterwards, (enzyme lives 7.8 * 10 to add 0.3～0.5% pepsin
5U/g), 40～60 ℃ of stirred in water bath enzymolysis 3～10 hours, keep pH simultaneously in above-mentioned scope.Transfer pH7～9 with 0.5 mol potassium hydroxide again, (enzyme lives 5 * 10 to add 0.3～0.5% trypsase
4U/g) 45～55 ℃ of stirred in water bath enzymolysis 2～7 hours, keep pH simultaneously in above-mentioned scope.Speed is chilled to below 30 ℃ behind the enzymolysis, keeps cessation reaction 30 minutes.To neutral, high speed centrifugation (6000 rev/mins) separated 15 minutes with 6 mol hydrochloric acid adjust pHs, and sediment is insoluble hydrolysate, collected mother liquor.The same single enzymolysis of remaining operation.
3. external source enzyme resolving after the self-dissolving
The I self-dissolving:
A puts into the self-dissolving 24～48 hours in pH6～7.5, under 0～4 ℃ of temperature of aseptic enzymatic vessel with sea cucumber milk;
Or b puts into aseptic enzymatic vessel with sea cucumber milk, in pH6～7.5, normal temperature self-dissolving 6～8 hours;
Or c with sea cucumber milk with ultraviolet ray irradiation 10～30 minutes after, with concentration be the sodium chloride of 0.06～0.08 mol in pH7.0～7.5, temperature is that the gradient temperature of 40～65 ℃ (5 ℃ of per minute risings) is handled.
The II external source enzyme resolving: will transfer pH to 1～5 from solution with 6 mol hydrochloric acid through what one of above-mentioned three kinds of self-dissolving methods obtained, adding 0.3～0.5% pepsin was 40～60 ℃ of enzymolysis 1～2 hour.Speed is chilled to below 30 ℃ behind the enzymolysis, keeps cessation reaction 30 minutes.To neutral, high speed centrifugation separates with 0.5 mol potassium hydroxide adjust pH, and sediment is insoluble hydrolysate, collects mother liquor.The same single enzymolysis of remaining operation.
4. autolytic enzyme is as the exogenous enzymes enzymolysis that combines with exogenous enzymes
I autolytic enzyme extracting method: fresh sea cucumber intestine tissue is smashed to pieces, be dissolved in 2～15 times 0.1M phosphate buffer (pH6～7.5), left standstill extracting 2～13 hours.Centrifugal afterwards, remove precipitation, in supernatant, slowly add solid ammonium sulfate to 10～30% saturation degree, make it dissolving, to saltout 2～24 hours, refrigerated centrifuge goes precipitation.In supernatant, add solid ammonium sulfate to 80～95% saturation degree, saltoutd 6～48 hours, dialysed afterwards 6～48 hours, centrifugal, get precipitation, this precipitation is dissolved in the 0.1M phosphate buffer (pH6～7.5), dialysed refrigerated centrifuge once more 6～48 hours, must precipitate, above process is all carried out under 10 ℃, will precipitate freeze drying and get the autolytic enzyme powder, also this enzyme powder can be used G-25 gel column purifying.
II autolytic enzyme enzymolysis: in sea cucumber milk, add 0.3～3% above-mentioned autolytic enzyme powder, in pH6～8,15～30 ℃ of following enzymolysis of temperature 3～8 hours,
The III external source enzyme resolving: transfer pH1～5 with 6 mol hydrochloric acid, add 0.3～0.5% pepsin, in 40～60 ℃ of enzymolysis 1～2 hour, speed was chilled to below 30 ℃ behind the enzymolysis, keeps cessation reaction 30 minutes.To neutral, high speed centrifugation separates with 0.5 mol potassium hydroxide adjust pH, and sediment is insoluble hydrolysate, collects mother liquor, the same single enzymolysis of remaining operation.
3, purifying: thick polysaccharide is dissolved in 1: 10～50 (grams per milliliter) water, and the centrifugal insoluble matter of removing adds the solid potassium acetate in supernatant, and making its concentration in solution is 0.5～4 mol, and refrigeration below 10 ℃ is spent the night.Collecting precipitation below 10 ℃.With the water of half volume last time it is dissolved again, transfer pH to 9～11, keep 45～55 ℃ of dropping H with 0.5 mol potassium hydroxide
2O
2The sugared at the most hydrolyzate that decolours is the light yellow terminal point that is.The centrifugal insoluble matter of removing in cooling back, clear liquid is cooled to below 4 ℃, transfer pH to 1.5～2.5 with 6 mol hydrochloric acid, the centrifugal precipitation of removing adds the solid potassium acetate in the clear liquid, make its concentration in solution reach 0.5～4 mol, refrigeration is spent the night, and collecting precipitation below 10 ℃ washs with ethanol, the acetone dehydration, the dry pure product of mucopolysaccharide that get.
Two, with the sea cucumber mucopolysaccharide be base-material, the preparation functional health care food.
1, the preparation of sea cucumber wine: sea cucumber mucopolysaccharide is dissolved in 4～60 degree wine, makes its concentration reach 0.01～1000 mg/litre.
2, the preparation of electuary:
1. mix: sea cucumber mucopolysaccharide base-material and additives mixed, base-material accounts for mixture 0.05～20%, these additives can be the Chinese medicine extracts with health-care effect, also can be the have additional nutrients dairy products that are worth, egg, vegetable protein etc., can also be white granulated sugar, honey and the citric acid etc. that improve taste.
2. centrifugal, filter: the new insoluble matter of removing most of residue and producing because of mixing.Before centrifugal, to handle, so that the precipitation that produces because of mixing forms as early as possible through heat exchanger and cool exchanger.Before concentrating, also to remove the less suspended particulate of density with flame filter press.
3. concentrate: adopt vacuum 86.25～98.64Kpa vacuum to concentrate earlier, temperature is lower than 50 ℃, and vacuum concentrates can accelerate the speed that concentrates, and keeps the color and luster of product.When vacuum concentrate not only thickness but also not too sticking chamber wall, use that jacketed pan is uncovered to be concentrated instead, until its density 〉=1.0 gram per centimeters
3
4. mixing granulation: employed cane sugar powder and soluble starch need be through the levigate and processing of sieving.Concentrate: soluble starch: cane sugar powder=1: 1: 2.Use edible alcohol to regulate humidity, granulate with comminutor in the moderate back of abundant mixing and humidity in mixer.
5. dry, pack: after choosing grain finishes,, be lower than 4%, the material of package materials selection protection against the tide to moisture in 60～80 ℃ of oven dry.
3, the preparation of capsule
1. granulation: adopt microcapsules technology to carry out granulation and handle, promptly in extract, add mucopolysaccharide, add 0.5～3% cycloheptaamylose again, boiled 2～10 minutes, raw meat astringent taste residual in the hydrolysate is wrapped up by cycloheptaamylose, offending raw meat astringent taste is covered, can also make active ingredient and external environment in the hydrolysate isolated, prevent to produce disadvantageous quality and change, thereby effectively keep nutritional labeling and other physiological activator.
2. dry: can pass through vacuum freeze drying, the hydrolysate that is about to above-mentioned microencapsulation is put into freeze drier, controls temperature below-40 ℃, vacuum 133 * 10
-3Pa, being lower than 4% up to moisture gets final product, also can pass through spray-drying, adopt vacuum concentration equipment to concentrate earlier, control vacuum at 86.25～98.64Kpa, 45～65 ℃ of temperature, opening steam valve when concentrating slowly heats up, progressively gas clean-up when concentrated material solid content reaches 4～17%, is carried out spray-drying.The feed temperature that dusts is at 45～50 ℃, and the material pump pressure is controlled at 19.6～49Kpa, 60～68 ℃ of temperature of outgoing airs, 150～180 ℃ of EATs.
3. cool off, pack: sieve removes the block of desciccate, cool to room temperature under the environment of 50～60% relative humidity, and the product that cooling is good incapsulates.
Illustrate: among the present invention, the solvable or insoluble protein hydrolysate that enzymolysis obtains all can adopt the method identical with said method to prepare capsule.
4, the preparation of oral liquid
1. mix: with sea cucumber mucopolysaccharide base-material and additives mixed, base-material accounts for mixture 0.05～20%, and is to the explanation of additive, identical with the preparation of electuary.
2. clarification: add 0.1～0.5% pectin and 0～7% honey as flocculant, with the precipitation of suspended particulate and foreign protein in the acceleration solution.Honey also can be used as flavor enhancement, stirs after the adding, and cooling is naturally left standstill to make and precipitated fully.
3. filter: adopt centrifugal or press filtration method, getting clear liquid is mother liquor.
4. seasoning: with 0～60 times of mother liquor dilution, add 1～7% white granulated sugar (or 0.05～0.2% albumen sugar, 6～12% sucrose), 0.02～0.4% citric acid, 0.01～0.05% salt with the weight of diluting back solution, 0.1～0.2% vitamin C, 0.5～9% honey.
5. clarification: with diatomite pressure filtration, via hole diameter is 5 μ m and 0.05 μ m two-stage filtering with microporous membrane again, thoroughly removes tiny suspended particulate and bacterium in the nutrient solution.
6. sterilize, pack: adopt pasteurize (80 ℃, 30 minutes) back can.
Illustrate: if when being the feedstock production oral liquid with the solvable of enzymolysis or insoluble protein hydrolysate, though can adopt method same as described above, handle but must take off raw meat, promptly add the nitration mixture of 0～0.2% citric acid and 0～0.2% malic acid, handled 10～30 minutes in 80～100 ℃; Perhaps adding 0.2～0.4% glucose heated 10～30 minutes in 90～100 ℃ of water-baths.
5, the preparation of tablet
Sea cucumber mucopolysaccharide is mixed (method prepares with electuary) before compressing tablet, add 5～10% starch or starch filled dose, compressing tablet with other polysaccharide or Chinese herbal medicine.
The present invention has following advantage:
1, a spot of sea cucumber mucopolysaccharide is put together, make functional food, and then make it to give full play to effect anticancer and enhancing immunity with certain content.
2, the method that adopts the sea cucumber self-dissolving to combine with exogenous enzymes among the present invention reduces cost first, also reduces simultaneously and introduces the pollution of exogenous enzymes to product.
The specific embodiment
The preparation of example 1 sea cucumber wine
Get 10 kilograms of dried sea-cucumbers and steep and send out, vertically cut open along a side jaggy, remove foreign material in ginseng mouth and the chamber with cutter, clean in clear water.Then with tissue mashing machine with its body wall fragmentation, and add 50 liters of volume water, under 40 ℃ of conditions, add 80 gram solid carbonic acid potassium, stir hydrolysis down 1 hour.Transferring pH with 6 mol hydrochloric acid afterwards is 7, and remains pH in this scope in enzymolysis process.(enzyme lives 5 * 10 to add 30 gram trypsase
4U/g) maintain the temperature at 45～47 ℃, enzymolysis is 7 hours under the stirring condition.Speed is chilled to below 30 ℃ afterwards, keeps cessation reaction 30 minutes.PH is extremely neutral with 6 mol hydrochloric acid readjustment.Mother liquor is collected in (6000 rev/mins) centrifugation at a high speed.Under constantly stirring, slowly add 8 kilograms of 95% ethanol, low temperature (≤10 ℃) spends the night.Centrifugal collecting precipitation afterwards, and successively with the washing of 90% ethanol, the acetone dehydration gets the thick polysaccharide of sea cucumber in 40～70 ℃ of temperature dryings again.Thick polysaccharide is dissolved in 2 premium on currency, and the centrifugal insoluble matter of removing adds 490 gram potassium acetates in supernatant, and refrigeration (below 10 ℃) is spent the night.Again in collecting precipitation below 10 ℃, with 1 liter water it is dissolved again.Transfer pH to 9 with 0.5 mol potassium hydroxide, keep 50 ℃ of sugared at the most hydrolyzates of dropping hydrogen peroxide decolouring to be the light yellow terminal point that is.The centrifugal insoluble matter of removing in cooling back, clear liquid is chilled to about 0 ℃, transfers pH to 1.5 with 6 mol hydrochloric acid, and the centrifugal precipitation of removing adds 245 grams per liter potassium acetates in the clear liquid, and refrigeration below 10 ℃ is spent the night.Collecting precipitation below 10 ℃, afterwards with ethanol washing, acetone dehydration, the dry pure product of sea cucumber mucopolysaccharide that get.The pure product of mucopolysaccharide of getting 1 gram extraction are dissolved in 1 liter 60 and spend in the wine, and the product that obtains is the sea cucumber wine that contains mucopolysaccharide.
The preparation of example 2 sea cucumber wines
Get 10 kilograms of dried sea-cucumbers and steep and send out, vertically cut open along a side jaggy, remove foreign material in ginseng mouth and the chamber with cutter, clean in clear water.Then with tissue mashing machine with its body wall fragmentation, and add 50 liters of volume water, under 40 ℃ of conditions, add 20 gram solid carbonic acid potassium, stir hydrolysis down 5 hours.Transferring pH with 6 mol hydrochloric acid afterwards is 9, and remains pH in this scope in enzymolysis process.(enzyme lives 5 * 10 to add 50 gram trypsase
4U/g), maintain the temperature at 53～55 ℃, enzymolysis is 2 hours under the stirring condition.Speed is chilled to below 30 ℃ afterwards, keeps cessation reaction 30 minutes.PH is extremely neutral with 6 mol hydrochloric acid readjustment.Mother liquor is collected in (6000 rev/mins) centrifugation at a high speed.Under constantly stirring, slowly add 3 kilograms of 95% ethanol, low temperature (≤10 ℃) spends the night.Centrifugal collecting precipitation afterwards, and successively with the washing of 100% ethanol, the acetone dehydration, again in 86.25～98.64Kpa vacuum dry the thick polysaccharide of sea cucumber.Thick polysaccharide is dissolved in 10 premium on currency, and the centrifugal insoluble matter of removing adds 784 gram potassium acetates in supernatant, and refrigeration (below 10 ℃) is spent the night.Again in collecting precipitation below 10 ℃, with 5 liters water it is dissolved again.Transfer pH to 11 with 0.5 mol potassium hydroxide, keep 50 ℃ of sugared at the most hydrolyzates of dropping hydrogen peroxide decolouring to be the light yellow terminal point that is.The centrifugal insoluble matter of removing in cooling back, clear liquid is cooled to about 0 ℃, transfers pH to 2.5 with 6 mol hydrochloric acid, and the centrifugal precipitation of removing adds 392 grams per liter potassium acetates in the clear liquid, and refrigeration below 10 ℃ is spent the night.Collecting precipitation below 10 ℃, afterwards with ethanol washing, acetone dehydration, the dry pure product of sea cucumber mucopolysaccharide that get.The pure product of mucopolysaccharide of getting 0.01 milligram of extraction are dissolved in 1 liter 4 and spend in the wine, and the product that obtains is the sea cucumber wine that contains mucopolysaccharide.
The preparation of example 3 sea cucumber mucopolysaccharide electuaries
Get 100 kilograms of salt marsh sea cucumbers, vertically cut open along a side jaggy, remove foreign material in ginseng mouth and the chamber, in clear water, clean with cutter.Then with tissue mashing machine with its body wall fragmentation, and add 500 premium on currency, under 40 ℃ of conditions, add 200 gram solid carbonic acid potassium, stir hydrolysis down 5 hours.Transferring pH with 6 mol hydrochloric acid afterwards is 1, and (enzyme lives 7.8 * 10 to add 500 gram pepsins
5U/g) 40 ℃ of stirred in water bath enzymolysis 10 hours, keep pH simultaneously in above-mentioned scope.Transfer pH to 7 with 0.5 mol potassium hydroxide afterwards, (enzyme lives 5 * 10 to add 500 gram trypsase
5U/g) 45 ℃ of stirred in water bath enzymolysis 2 hours, and keep pH at the further enzymolysis of above-mentioned scope, speed is chilled to below 30 ℃ behind the enzymolysis, keeps cessation reaction 30 minutes.With 6 mol hydrochloric acid pH is transferred to neutrality, (6000 rev/mins) centrifugation is 15 minutes at a high speed, collects mother liquor.Under constantly stirring, slowly add 30 kilograms of ethanol, remaining operation is identical with example 1.Thick polysaccharide is dissolved in 60 premium on currency, and the centrifugal insoluble matter of removing adds 5880 gram potassium acetates in supernatant, spend the night below 10 ℃.Collecting precipitation below 10 ℃ dissolves it with 30 premium on currency again.Transfer pH to 9 with 6 mol potassium hydroxide, keep 50 ℃ of sugared at the most hydrolyzates of dropping hydrogen peroxide decolouring to be the light yellow terminal point that is, the centrifugal insoluble matter of removing in cooling back, clear liquid is cooled to about 0 ℃, transfer pH to 1.5 with 6 mol hydrochloric acid, the centrifugal precipitation of removing adds 2940 gram potassium acetates in the clear liquid, spend the night below 10 ℃.Collecting precipitation below 10 ℃, ethanol washing afterwards, acetone dehydration, drying gets the pure product of polysaccharide.
Other gets 4 kilograms of fruits of Chinese wolfberry, 4 kilograms of rhodiola roots add 40 kg of water and boil, slow fire kept 1 hour, cross leaching filtrate, be concentrated into 36 liters, in concentrate, add 95 liters of ethanol, leave standstill after 10 hours and filter, with the way of distillation with ethanol steam extract, get the 12.5 gram sea cucumber mucopolysaccharides that obtain by said method, mix with 25 kilograms of extracts that obtain by said method, centrifugal after heat exchanger and cool exchanger processing, remove the less suspended particulates of density with flame filter press again, adopting vacuum afterwards is that 86.25～96.64Kpa vacuum concentrates, and temperature is lower than 50 ℃.When vacuum concentrate not only thickness but also not too sticking chamber wall, use that jacketed pan is uncovered to be concentrated instead, be 1.0 gram per centimeters until its density
3Afterwards concentrate, soluble starch and cane sugar powder are mixed with 1: 1: 2 ratio, regulate humidity with edible alcohol, in mixer, fully mix and humidity moderate after, granulate with comminutor.After granulation finishes, under 60～80 ℃ of conditions, dry, use In Aluminium Foil Packing at last to moisture 3.8%.
The preparation of example 4 sea cucumber mucopolysaccharide electuaries
Get 100 kilograms of salt marsh sea cucumbers, vertically cut open along a side jaggy, remove foreign material in ginseng mouth and the chamber, in clear water, clean with cutter.Then with tissue mashing machine with its body wall fragmentation, and add 400 premium on currency, under 70 ℃ of conditions, add 800 gram solid carbonic acid potassium, stir hydrolysis down 1 hour.Transfer pH to 5 with 6 mol hydrochloric acid afterwards, (enzyme lives 7.8 * 10 to add 300 gram pepsins
5U/g) 60 ℃ of stirred in water bath enzymolysis 3 hours, keep pH simultaneously in above-mentioned scope.Transfer pH to 9 with 0.5 mol potassium hydroxide afterwards, (enzyme lives 5 * 10 to add 300 gram trypsase
5U/g) 55 ℃ of stirred in water bath enzymolysis 7 hours, and keep pH at the further enzymolysis of above-mentioned scope, speed is chilled to below 30 ℃ behind the enzymolysis, keeps cessation reaction 30 minutes.With 6 mol hydrochloric acid pH is transferred to neutrality, (6000 rev/mins) centrifugation is 15 minutes at a high speed, collects mother liquor.Under constantly stirring, slowly add 80 kilograms of ethanol, remaining operation is identical with example 2.Thick polysaccharide is dissolved in 60 premium on currency centrifugal insoluble matter of removing.In supernatant, add 5880 gram potassium acetates, spend the night below 10 ℃.Collecting precipitation below 10 ℃, with 30 premium on currency it is dissolved again, transfer pH to 11 with 6 mol potassium hydroxide, keep 50 ℃ of sugared at the most hydrolyzates of dropping hydrogen peroxide decolouring to be the light yellow terminal point that is, the centrifugal insoluble matter of removing in cooling back, clear liquid is cooled to about 0 ℃, with 6 mol hydrochloric acid adjust pHs to 2.5, the centrifugal precipitation of removing adds 2940 gram potassium acetates in the clear liquid, spend the night below 10 ℃.Collecting precipitation below 10 ℃, ethanol washing afterwards, acetone dehydration, drying gets the pure product of polysaccharide.
Other gets 4 kilograms of dates, 4 kilograms of longan add 40 kg of water and boil, slow fire kept 1 hour, cross leaching filtrate, be concentrated into 20 liters, in concentrate, add 95 liters of ethanol, leave standstill filtration in 10 hours, with the way of distillation with ethanol steam extract, get 3 kilograms of sea cucumber mucopolysaccharides that obtain by said method, mix with 15 kilograms of extracts that obtain by said method, centrifugal after heat exchanger and cool exchanger processing, remove the less suspended particulates of density with flame filter press again, adopting vacuum afterwards is that 86.25～96.64Kpa vacuum concentrates, and temperature is lower than 50 ℃.When vacuum concentrate not only thickness but also not too sticking chamber wall, use that jacketed pan is uncovered to be concentrated instead, be 1.0 gram per centimeters until its density
3Afterwards concentrate, soluble starch and cane sugar powder are mixed with 1: 1: 2 ratio, regulate humidity with edible alcohol, in mixer, fully mix and humidity moderate after, granulate with comminutor.After granulation finishes, under 60～80 ℃ of conditions, dry, use In Aluminium Foil Packing at last to moisture 4%.
The preparation of example 5 sea cucumber mucopolysaccharide oral liquids
Get 40 kilograms of bright sea cucumbers, remove internal organ, remove muscle, clean.Then with tissue mashing machine with its body wall fragmentation, and add 200 liters water, be that 6 normal temperature were placed 6 hours at pH, make the sea cucumber self-dissolving, transferring pH with 6 mol hydrochloric acid afterwards is 1, adds 120 gram pepsins, in 40～45 ℃ of enzymolysis 2 hours, speed is chilled to below 30 ℃ behind the enzymolysis, keeps cessation reaction 30 minutes, with 0.5 mol potassium hydroxide, return adjust pH to neutral, remaining operation is identical with example 1, gets the pure product of mucopolysaccharide.
Other gets 1.6 kilograms of rhodiola roots, 1.6 kilograms of dates add 16 kg of water and boil, slow fire kept 1 hour, cross leaching filtrate, be concentrated into 8 liters, adding 95 liters of ethanol in concentrate leaves standstill, with the way of distillation with ethanol steam extract, get 1.2 kilograms of sea cucumber mucopolysaccharides that obtain by said method, mix with 6 kilograms of extracts that obtain by said method.Add 30 gram pectin and stir, cooling was left standstill 2 hours naturally, made precipitation fully.Adopt centrifugal (4000 rev/mins, 15 minutes) get supernatant, with 60 times of this liquid dilutions, add 4.32 kilograms of white sugar, 86.4 gram citric acids, 43.2 gram salt, 432 gram vitamin Cs, 2.16 gram honey mixings, with diatomite pressure filtration, via hole diameter is 5 μ m and 0.5 μ m two-stage filtering with microporous membrane again.Again filtrate is carried out 80 ℃, 30 minutes pasteurize, after get final product can.
The preparation of example 6 sea cucumber mucopolysaccharide oral liquids
Get 40 kilograms of bright sea cucumbers, remove internal organ, remove muscle, clean.Then with tissue mashing machine with its body wall fragmentation, be that 6 normal temperature were placed 8 hours in the pH value, make the sea cucumber self-dissolving, transferring pH with 6 mol hydrochloric acid afterwards is 5, add 150 gram pepsins, in 55～60 ℃ of enzymolysis 1 hour, speed was chilled to below 30 ℃ behind the enzymolysis, kept 30 minutes, cessation reaction with 0.5 mol potassium hydroxide, is returned adjust pH to neutral, remaining operation is identical with example 2, gets the pure product of mucopolysaccharide.
Other gets 1.6 kilograms of fruits of Chinese wolfberry, 1.6 kilograms of longan add 16 kg of water and boil, slow fire kept 1 hour, cross leaching filtrate, be concentrated into 15 liters, adding 95 liters of ethanol in concentrate leaves standstill, with the way of distillation with ethanol steam extract, get the 6 gram sea cucumber mucopolysaccharides that obtain by said method, mix with 12 kilograms of extracts that obtain by said method.Add 12 gram pectin, 0.84 kilogram of honey and stir, cooling was left standstill 3 hours naturally, made precipitation complete.Adopt press filtration, get filtrate, add 0.91 kilogram of white sugar, 52 gram citric acids, 6.5 gram salt, 26 gram vitamin Cs, 1.17 gram honey mixings, with diatomite pressure filtration, via hole diameter is 5 μ m and 0.5 μ m two-stage filtering with microporous membrane again.Again filtrate is carried out 80 ℃, 30 minutes pasteurize, after get final product can.
The preparation of example 7 beche-de-mer capsules
Get 40 kilograms of fresh sea cucumbers, remove internal organ, remove muscle, clean.Then with tissue mashing machine with its body wall fragmentation, and add 80 liters water, beat even.With ultraviolet ray irradiation 10 minutes, is 0.06 mol sodium chloride at pH be 7 with concentration then with this sea cucumber milk, and temperature is to carry out the self-dissolving processing under the gradient temperature condition of 40～65 ℃ (raising 5 ℃ in per 3 minutes).Transferring pH with 6 mol hydrochloric acid afterwards is 1, adds 160 gram pepsin enzymolysis 2 hours, and speed is chilled to below 30 ℃ behind the enzymolysis, keeps cessation reaction 30 minutes.Remaining operation is identical with example 1, obtains the pure product of sea cucumber mucopolysaccharide.
Other gets 80 kilograms in date, longan adds 800 liters in water for 80 kilograms, repeats the operation of preparation extract in the example 3, gets 160 liters of extracts, add 80 gram mucopolysaccharides, add 800 gram cycloheptaamyloses again, boiled 2 minutes, put into the freeze drier drying, control temperature below-40 ℃, vacuum 133 * 10
-3Pa stops when moisture 4%.Sieve removes block afterwards, is cooled to room temperature under the environment of 50% relative humidity, and the product that cooling is good incapsulates.
The preparation of example 8 beche-de-mer capsules
Get 40 kilograms of fresh sea cucumbers, remove internal organ, remove muscle, clean.Then with tissue mashing machine with its body wall fragmentation, and add 120 liters water, beat even.With ultraviolet ray irradiation 30 minutes, is 0.08 mol sodium chloride at pH be 7.5 with concentration then with this sea cucumber milk, and temperature is to carry out the self-dissolving processing under the gradient temperature condition of 40～65 ℃ (raising 5 ℃ in per 3 minutes).Transferring pH with 6 mol hydrochloric acid afterwards is 5, adds 180 gram pepsin enzymolysis 1 hour, and speed is chilled to below 30 ℃ behind the enzymolysis, keeps cessation reaction 30 minutes.Remaining operation is identical with example 2, obtains the pure product of sea cucumber mucopolysaccharide.
Other gets 80 kilograms of the fruits of Chinese wolfberry, rhodiola root adds 800 liters in water for 80 kilograms, repeats the operation of preparation extract in the example 3, gets 160 liters of extracts, add 80 gram mucopolysaccharides, add 4800 gram cycloheptaamyloses again, boiled 10 minutes, control vacuum is 45～65 ℃ of 86.25～98.64Kpa, temperature, concentrate, when solid content is 4% in concentrating material, the beginning spray-drying, 45～50 ℃ of feed temperatures dust, material pump pressure 19.6～49.0Kpa, 60～68 ℃ of temperature of outgoing airs, 150～180 ℃ of EATs.Sieve removes block afterwards, is cooled to room temperature under the environment of 60% relative humidity, and the product that cooling is good incapsulates.
The preparation of example 9 sea cucumber mucopolysaccharide tablets
200 kilograms of fresh sea cucumber intestine tissues are smashed to pieces, be dissolved in 400 liters of 0.1M sodium phosphate buffers (pH6.0～7.5), left standstill extracting 13 hours, the centrifugal afterwards precipitation of removing, in supernatant, add solid ammonium sulfate to 80% saturation degree, saltoutd 48 hours, the 6 hours centrifuging and taking precipitations of dialysing afterwards are dissolved in precipitation in the 0.1M phosphate buffer solution (pH6.0～7.5), dialysed once more 48 hours, centrifugal must the precipitation, more than operation is all being carried out below 10 ℃, will precipitate freeze drying and get autolytic enzyme enzyme powder.
Other gets the dried sea-cucumber that 50 kilograms of bubbles are sent out, and vertically cuts open along a side jaggy with cutter, removes foreign material in ginseng mouth and the chamber, and is clean in clear water, then with tissue mashing machine with its body wall fragmentation, and add 250 premium on currency and beat even.Stirring condition adds the autolytic enzyme that 150 grams are extracted by said method down, enzymolysis is 8 hours under pH6,15 ℃ of conditions of temperature, with 6 mol hydrochloric acid pH is transferred to 1 afterwards, add 150 gram pepsins, enzymolysis is 2 hours under 40～45 ℃ of conditions, speed is chilled to below 30 ℃ behind the enzymolysis, keeps cessation reaction 30 minutes.Remaining operation is identical with example 1, gets the pure product of mucopolysaccharide.Get 100 gram mucopolysaccharides, be added in the extract that makes by 400 gram rhodiola roots, 200 gram dates, 200 gram longan and 2000 ml waters, add 5 gram cycloheptaamyloses then, boiled 10 minutes, send into drying in the freeze drier again, control temperature below-40 ℃, vacuum 133 * 10
-3Pa is 4% termination up to moisture.Get the above-mentioned dry powder of 300 grams add 30 gram starch through the tablet press machine compressing tablet get final product the sea cucumber mucopolysaccharide sheet.
The preparation of example 10 sea cucumber mucopolysaccharide tablets
200 kilograms of fresh sea cucumber intestine tissues are smashed to pieces, be dissolved in 3000 liters of 0.1M sodium phosphate buffers (pH6.0～7.5), left standstill extracting 2 hours, the centrifugal afterwards precipitation of removing, in supernatant, add solid ammonium sulfate to 95% saturation degree, saltoutd 6 hours, the 48 hours centrifuging and taking precipitations of dialysing afterwards are dissolved in precipitation in the 0.1M phosphate buffer solution (pH6.0～7.5), dialysed once more 6 hours, centrifugal must the precipitation, more than operation is all being carried out below 10 ℃, will precipitate freeze drying and get autolytic enzyme enzyme powder.
Other gets the dried sea-cucumber that 50 kilograms of bubbles are sent out, and vertically cuts open along a side jaggy with cutter, removes foreign material in ginseng mouth and the chamber, and is clean in clear water, then with tissue mashing machine with its body wall fragmentation.Stirring condition adds the autolytic enzyme that 1500 grams are extracted by said method down, at pH8, enzymolysis is 3 hours under 30 ℃ of conditions of temperature, with 6 mol hydrochloric acid pH is transferred to 5 afterwards, add 250 gram pepsins, enzymolysis is 1 hour under 55～60 ℃ of conditions, and speed is chilled to below 30 ℃ behind the enzymolysis, kept cessation reaction 30 minutes.Remaining operation is identical with example 2, gets the pure product of mucopolysaccharide.Get 100 gram mucopolysaccharides, be added in the extract that makes by 400 gram rhodiola roots, 200 gram dates, 200 gram longan and 2000 ml waters, add 30 gram cycloheptaamyloses then, boiled 2 minutes, send into drying in the freeze drier again, control temperature below-40 ℃, vacuum 133 * 10
-3Pa is 4% termination up to moisture.Get the above-mentioned dry powder of 300 grams add 15 gram starch through the tablet press machine compressing tablet get final product the sea cucumber mucopolysaccharide sheet.
The preparation of example 11 sea cucumber proteolysis juice beverages
Get 40 kilograms of fresh sea cucumbers, remove internal organ, remove muscle, clean.Then with tissue mashing machine with its body wall fragmentation, and add 200 liters water.Was 6 with sea cucumber milk at pH, and temperature is under 4 ℃ of conditions, and self-dissolving is 24 hours in aseptic enzymatic vessel, and transferring pH with 6 mol hydrochloric acid afterwards is 5, adds 200 gram pepsins, in 40 ℃ of enzymolysis 2 hours.Speed is chilled to below 30 ℃ behind the enzymolysis, keeps cessation reaction 30 minutes.To neutral, mother liquor is collected in (6000 rev/mins) centrifugation at a high speed with 0.5 mol potassium hydroxide readjustment pH, slowly adds 32 kilogram of 95% ethanol under constantly stirring, and spends the night in low temperature below 10 ℃.The centrifugal afterwards supernatant that gets carries out decompression distillation and reclaims ethanol, with the liquid filtering of remainder, adopts 95～100 ℃ to continue 2 minutes, and it is standby to sterilize.Insert Bulgarian Lactobacillus then, bacterium seed liquor inoculum concentration 120 grams, 30 ℃ of fermentation temperatures.Add 120 gram skimmed milk powers and 120 gram glucose simultaneously at pH4.0, during acidity 0.4%, cool the temperature to below 6 ℃, stop fermentation.In zymotic fluid, add 120 gram white granulated sugar mixings, and through the pressure homogeneous of high pressure homogenizer with 20Mpa.To be heated to 80 ℃ through the zymotic fluid that said process makes, bottling while hot, sealing is cooled to room temperature again after pasteurize (80 ℃, 30 minutes).
The preparation of example 12 sea cucumber mucopolysaccharide tablets
Get 40 kilograms of fresh sea cucumbers, remove internal organ, remove muscle, clean.Then with tissue mashing machine with its body wall fragmentation, be 7.5 with sea cucumber milk at pH, temperature is under 0 ℃ of condition, self-dissolving is 48 hours in aseptic enzymatic vessel, transferring pH with 6 mol hydrochloric acid afterwards was 1, adds 120 gram pepsins, in 60 ℃ of enzymolysis 1 hour.Speed is chilled to below 30 ℃ behind the enzymolysis, keeps cessation reaction 30 minutes.To neutral, mother liquor is collected in (6000 rev/mins) centrifugation at a high speed with 0.5 mol potassium hydroxide readjustment pH, slowly adds 12 kilogram of 95% ethanol under constantly stirring, and spends the night in low temperature below 10 ℃.The centrifugal afterwards supernatant that gets carries out decompression distillation and reclaims ethanol, with the liquid filtering of remainder, adopts the instantaneous heating of superhigh temperature, and it is standby to sterilize.Insert Bulgarian Lactobacillus then, bacterium seed liquor inoculum concentration 40 grams, 50 ℃ of fermentation temperatures.Add 40 gram skimmed milk powers and 20 gram glucose simultaneously,, during acidity 0.2%, cool the temperature to below 6 ℃, stop fermentation at pH4.5.In zymotic fluid, add 40 gram white granulated sugar mixings, and through the pressure homogeneous of high pressure homogenizer with 40Mpa.To be heated to 80 ℃ through the zymotic fluid that said process makes, bottling while hot, sealing is cooled to room temperature again after pasteurize (80 ℃, 30 minutes).
Annotate: percentage used in the present specification all is weight percentage.
Claims (3)
1, a kind of preparation method of sea cucumber mucopolysaccharide is characterized in that:
1. raw material and processing: the learn from else's experience dried sea-cucumber handled or salt marsh sea cucumber or bright sea cucumber are added the water of 0～5 times of volume, with tissue mashing machine with its body wall fragmentation,
2. extract sea cucumber mucopolysaccharide
The a self-dissolving: self-dissolving 24～48 hours under 0～4 ℃ the condition with sea cucumber milk in pH6～7.5, temperature,
The b enzymolysis: will transfer to 1～5 from pH value of solution with 6 mol hydrochloric acid, and add 0.3～0.5% pepsin, enzymolysis is 1～2 hour under 40～60 ℃ of conditions, and speed is as cold as below 30 ℃ behind the enzymolysis, kept 30 minutes,
C separates: transfer enzymolysis liquid pH to neutral with 0.5 mol potassium hydroxide, high speed centrifugation separates, and collects mother liquor,
D alcohol is molten: stir that slowly to add mass percent down be 30～80% 95% ethanol ,≤10 ℃ are spent the night,
E purifies: centrifugal collecting precipitation, and, in 40～70 ℃ of dryings or vacuum drying, get the sea cucumber mucopolysaccharide crude product after the acetone dehydration successively with the washing of 90～100% ethanol,
3. purifying: with the mucopolysaccharide dissolving crude product in 1: 10～50 (g/ml) water, centrifugal, in clear liquid, add the solid potassium acetate, making its concentration in solution is 0.5～4 mol, refrigeration is spent the night, the low temperature collecting precipitation, and using last time again, the water of half volume dissolves it, transfer pH to 9～11 with 0.5 mol potassium hydroxide, keep 45～55 ℃ to drip H
2O
2The sugared at the most hydrolyzate that decolours is the light yellow terminal point that is, cooling is centrifugal, supernatant is cooled to below 4 ℃, transfer pH1.5～2.5 with 6 mol hydrochloric acid, centrifugal, in supernatant, add the solid potassium acetate, make its concentration in solution reach 0.5～4 mol, refrigeration is spent the night, low temperature collecting precipitation, ethanol washing afterwards, the acetone dehydration, the dry pure product of mucopolysaccharide that get.
2, a kind of preparation method of sea cucumber mucopolysaccharide is characterized in that:
1. raw material and processing: the learn from else's experience dried sea-cucumber handled or salt marsh sea cucumber or bright sea cucumber are added the water of 0～5 times of volume, with tissue mashing machine with its body wall fragmentation,
2. extract sea cucumber mucopolysaccharide
The a self-dissolving: with sea cucumber milk self-dissolving 6～8 hours under pH6～7.5 normal temperature,
The b enzymolysis: will transfer to 1～5 from pH value of solution with 6 mol hydrochloric acid, and add 0.3～0.5% pepsin, enzymolysis is 1～2 hour under 40～60 ℃ of conditions, and speed is as cold as below 30 ℃ behind the enzymolysis, kept 30 minutes,
C separates: transfer enzymolysis liquid pH to neutral with 0.5 mol potassium hydroxide, high speed centrifugation separates, and collects mother liquor,
D alcohol is molten: stir that slowly to add mass percent down be 30～80% 95% ethanol ,≤10 ℃ are spent the night,
E purifies: centrifugal collecting precipitation, and, in 40～70 ℃ of dryings or vacuum drying, get the sea cucumber mucopolysaccharide crude product after the acetone dehydration successively with the washing of 90～100% ethanol,
3. purifying: with the mucopolysaccharide dissolving crude product in 1: 10～50 (g/ml) water, centrifugal, in clear liquid, add the solid potassium acetate, making its concentration in solution is 0.5～4 mol, refrigeration is spent the night, the low temperature collecting precipitation, and using last time again, the water of half volume dissolves it, transfer pH to 9～11 with 0.5 mol potassium hydroxide, keep 45～55 ℃ to drip H
2O
2The sugared at the most hydrolyzate that decolours is the light yellow terminal point that is, cooling is centrifugal, supernatant is cooled to below 4 ℃, transfer pH1.5～2.5 with 6 mol hydrochloric acid, centrifugal, in supernatant, add the solid potassium acetate, make its concentration in solution reach 0.5～4 mol, refrigeration is spent the night, low temperature collecting precipitation, ethanol washing afterwards, the acetone dehydration, the dry pure product of mucopolysaccharide that get.
3, a kind of preparation method of sea cucumber mucopolysaccharide is characterized in that:
1. raw material and processing: the learn from else's experience dried sea-cucumber handled or salt marsh sea cucumber or bright sea cucumber are added the water of 0～5 times of volume, with tissue mashing machine with its body wall fragmentation,
2. extract sea cucumber mucopolysaccharide
The a self-dissolving: with sea cucumber milk with ultraviolet ray irradiation 10～30 minutes after, the sodium chloride that with concentration is 0.06～0.08 mol is to handle under the condition of gradient temperature of 40～65 ℃ (5 ℃ of risings in per 3 minutes) in pH7.0～7.5, temperature,
The b enzymolysis: will transfer to 1～5 from pH value of solution with 6 mol hydrochloric acid, and add 0.3～0.5% pepsin, enzymolysis is 1～2 hour under 40～60 ℃ of conditions, and speed is as cold as below 30 ℃ behind the enzymolysis, kept 30 minutes,
C separates: transfer enzymolysis liquid pH to neutral with 0.5 mol potassium hydroxide, high speed centrifugation separates, and collects mother liquor,
D alcohol is molten: stir that slowly to add mass percent down be 30～80% 95% ethanol ,≤10 ℃ are spent the night,
E purifies: centrifugal collecting precipitation, and, in 40～70 ℃ of dryings or vacuum drying, get the sea cucumber mucopolysaccharide crude product after the acetone dehydration successively with the washing of 90～100% ethanol,
3. purifying: with the mucopolysaccharide dissolving crude product in 1: 10～50 (g/ml) water, centrifugal, in clear liquid, add the solid potassium acetate, making its concentration in solution is 0.5～4 mol, refrigeration is spent the night, the low temperature collecting precipitation, and using last time again, the water of half volume dissolves it, transfer pH to 9～11 with 0.5 mol potassium hydroxide, keep 45～55 ℃ to drip H
2O
2The sugared at the most hydrolyzate that decolours is the light yellow terminal point that is, cooling is centrifugal, supernatant is cooled to below 4 ℃, transfer pH1.5～2.5 with 6 mol hydrochloric acid, centrifugal, in supernatant, add the solid potassium acetate, make its concentration in solution reach 0.5～4 mol, refrigeration is spent the night, low temperature collecting precipitation, ethanol washing afterwards, the acetone dehydration, the dry pure product of mucopolysaccharide that get.
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| CN102002110A (en) * | 2010-11-24 | 2011-04-06 | 刘全胜 | Preparation method of polysaccharides from sea cucumber |
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| CN110339399A (en) * | 2019-07-09 | 2019-10-18 | 上海市第六人民医院 | A kind of biomimetic artificial bone and preparation method thereof |
| CN115894734A (en) * | 2022-12-02 | 2023-04-04 | 大连深蓝肽科技研发有限公司 | Method for extracting sea cucumber polysaccharide from sea cucumber cooking liquor with high efficiency and low pollution |
| CN115894734B (en) * | 2022-12-02 | 2024-02-23 | 大连深蓝肽科技研发有限公司 | Method for extracting sea cucumber polysaccharide from sea cucumber cooking liquor with high efficiency and low pollution |
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