CN103951713A - Comprehensive utilization method for improving value of cucumis sativus seed medicinal materials - Google Patents
Comprehensive utilization method for improving value of cucumis sativus seed medicinal materials Download PDFInfo
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- CN103951713A CN103951713A CN201410211320.1A CN201410211320A CN103951713A CN 103951713 A CN103951713 A CN 103951713A CN 201410211320 A CN201410211320 A CN 201410211320A CN 103951713 A CN103951713 A CN 103951713A
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Abstract
The invention discloses a comprehensive utilization method for improving the value of cucumis sativus seed medicinal materials, which relates to a comprehensive utilization method for cucumis sativus seeds. The invention aims to solve the problems that in the prior art, various components in cucumis sativus seeds can not be simultaneously extracted and utilized, the extraction cost is high, and resources are wasted. The method comprises the following steps: 1, extracting water-soluble components in cucumis sativus seeds by using a water immersion method; and 2, after cucumis sativus seed filter residues obtained in the step 1 are dried, crushing the dried residues into powder, and carrying out extraction on the powder by using a supercritical CO2 extraction method so as to obtain cucumis sativus seed fatty oil and residue extracts; 3, carrying out enzymolysis on the residue extracts obtained in the steps by using an enzymolysis method, and carrying out inactivation and filtering on the obtained product; and finally, separating cucumis sativus seed polysaccharides and cucumis sativus seed peptides in filter liquor obtained after enzymolysis by using an ultrafiltration method. According to the invention, the comprehensive utilization value of cucumis sativus seed medicinal materials is effectively improved and a new way is provided for the utilization of water-soluble ingredients and residues of cucumis sativus seeds. The invention can obtain a comprehensive utilization method for improving the value of cucumis sativus seed medicinal materials.
Description
Technical field
The present invention relates to a kind of method of comprehensive utilization of Semen Cucumidis sativi, relate in particular to the method for comprehensive utilization of the Multiple components such as fatty oil in Semen Cucumidis sativi, polysaccharide, polypeptide, VITAMIN and organic acid, belong to the extraction of Semen Cucumidis sativi and utilize field.
Background technology
Semen Cucumidis sativi, different name Harry Soviet Union, is the dry mature seed of Curcurbitaceae Cucumis plant cucumber (Cucumis sativus L.), and all there is cultivation all parts of the country, and medicine source is wide, and pharmaceutical use is high.It is widely used in synthetism, strong bone and replenishes the calcium among the people, medication is with a long history, records Semen Cucumidis sativi and has reunion of fractured tendons and bones, dispels the wind, the effect of dissolving phlegm in the herbal > > of < < China.Cure mainly the fracture injury of tendon and muscle, rheumatic arthralgia, old asthma due to excessive phlegm.Modern study confirmation, Semen Cucumidis sativi fatty oil content is higher, wherein contains the trace elements such as lipid acid (oleic acid, linolic acid), sterols composition and Mg, Ca, Fe, P of a large amount of needed by human.In addition, Semen Cucumidis sativi also contains the various active compositions such as polysaccharide, polypeptide, organic acid and VITAMIN, the diseases such as the soreness of waist, backache, numbness in hands and feet, legs and feet cramp, rheumatosis, sacroiliitis, cervical spondylosis, fracture, bone split are had and well resumed treatment and nourishing function, can regulate the interaction between human internal organ, promote the regeneration of human body cell, reconcile channels and collaterals simultaneously, nutrition brain, cerebellum, make people's memory, the coordination of middleman's health and balance, belong to dietotherapeutic kind.But to the utilization of Semen Cucumidis sativi, be mainly lipid acid and trace element at present, relatively less to polysaccharide, polypeptide components utilising, and it is few to utilizing of other water soluble components such as VITAMIN and organic acid, the Semen Cucumidis sativi residue extracting after fatty oil generally abandons as refuse, causes the serious waste of resource.Therefore how comprehensive development and utilization Semen Cucumidis sativi, improves the comprehensive utilization ratio of resource, is one of current problem demanding prompt solution.
Summary of the invention
The object of the invention is to solve prior art and can not extract simultaneously and utilize the Multiple components in Semen Cucumidis sativi, the problem of extraction cost height and the wasting of resources, and a kind of method of comprehensive utilization that Semen Cucumidis sativi medicinal material is worth that improves is provided.
Improve the method for comprehensive utilization that Semen Cucumidis sativi medicinal material is worth, comprising:
One, adopt water seaoning to extract the water soluble component in Semen Cucumidis sativi;
Water seaoning described in step 1 comprises: Semen Cucumidis sativi is pulverized, obtained Semen Cucumidis sativi meal; Semen Cucumidis sativi meal is immersed in to 12h～24h in water, refilters, obtain Semen Cucumidis sativi filter residue and filtrate, in filtrate, contain aqueous soluble active constituent; Described aqueous soluble active constituent comprises VITAMIN and organic acid;
The solid-liquid ratio of the Semen Cucumidis sativi meal described in step 1 and water is 1:(5～10);
Two, Semen Cucumidis sativi filter residue step 1 being obtained is ground into powder after drying, then adopts supercritical CO
2extraction process extracts, and obtains Semen Cucumidis sativi fatty oil and residue extract remainder;
Supercritical CO described in step 2
2extraction process, comprises the following steps: at extracting pressure 25MPa～35MPa, 35 ℃～55 ℃ of extraction temperature and CO
2continuous extraction 1h～2h under flow 45L/min～60L/min condition, obtains Semen Cucumidis sativi fatty oil and residue extract remainder;
Three, the residue extract remainder that adopts enzymolysis process to obtain step 2 carries out enzymolysis, deactivation 10min～20min at the temperature of 80 ℃～100 ℃ again, in the enzymolysis solution after deactivation, add distilled water, speed with 5000r/min～10000r/min is carried out centrifugal, get supernatant liquor and filter, obtain the filtrate after enzymolysis; Finally use Semen Cucumidis sativi polysaccharide and Semen Cucumidis sativi polypeptide in the filtrate after the separated enzymolysis of ultrafiltration process;
Enzymolysis solution after deactivation described in step 3 and the volume ratio of distilled water are 1:1;
Enzymolysis process described in step 3, comprises the following steps: in the residue extract remainder obtaining to step 2, add distilled water, according to g/mL meter, the solid-liquid ratio of residue extract remainder and water is 1:6～1:20, and stirring and evenly mixing obtains residue extract remainder/distilled water mixed solution; Regulating the pH of residue extract remainder/distilled water mixed solution is 5.0～8.0, then adds proteolytic enzyme, in temperature, is to stir enzymolysis 3h～8h under 50 ℃～65 ℃ and the stirring velocity condition that is 200r/min～300r/min; The total amount that adds of described proteolytic enzyme is 2500U/g～5500U/g;
Semen Cucumidis sativi polysaccharide in filtrate after the separated enzymolysis of use ultrafiltration process described in step 3 and the method for Semen Cucumidis sativi polypeptide, comprise the following steps:
1. the ultra-filtration membrane ultrafiltration that is 10KDa by the filtrate after enzymolysis through trapped molecular weight, filtrate residual content in ultrafiltration cup after enzymolysis be filtrate cumulative volume after enzymolysis 1/5 time, collect filtered solution, and to supplementing the original volume of distilled water to the filtrate after enzymolysis, ultrafiltration again in ultrafiltration cup;
2. repeating step is 1. 3 times～4 times, stops ultrafiltration, collects each filtered solution and trapped fluid; Step 2. described each filtered solution is the polypeptide liquid of molecular mass <10KDa; The trapped fluid of step described in is 2. the Crude polysaccharides liquid of molecular mass >10KDa;
3. under temperature is the condition of 50 ℃～60 ℃, trapped fluid is evaporated to 1/2～1/3 of trapped fluid initial volume, in the trapped fluid after concentrated, adds ethanol, obtain trapped fluid/ethanolic soln; By the standing 8h～12h of trapped fluid/ethanolic soln, suction filtration, is precipitated, and uses successively dehydrated alcohol, anhydrous diethyl ether and washing with acetone precipitation, and vacuum-drying or lyophilize, obtain Semen Cucumidis sativi polysaccharide powder; In the trapped fluid/ethanolic soln of step described in 3., the volume of ethanol accounts for 60%～95% of trapped fluid/ethanolic soln cumulative volume;
4. the polypeptide liquid of the molecular mass <10KDa 3. step being collected is the ultra-filtration membrane ultrafiltration of 1KDa and 650Da through molecular weight cut-off successively, collect respectively trapped fluid and filtered solution, obtain the polypeptide solution that molecular mass is respectively 1KDa～10KDa, 650Da～1KDa and <650Da, respectively three peptide species solution for vacuum are dried or lyophilize, obtain the Semen Cucumidis sativi polypeptide powder of three kinds of differing molecular quality.
Advantage of the present invention: one, the present invention can extract simultaneously and utilize the number of chemical compositions such as fatty oil in Semen Cucumidis sativi, polysaccharide, polypeptide, VITAMIN and organic acid, reaches the comprehensive utilization of Semen Cucumidis sativi, has saved resource;
Two, extraction process of the present invention is simple, and extraction conditions is gentle, is conducive to retain the biological activity of its chemical composition, safety and environmental protection;
Three, the Semen Cucumidis sativi water soluble component that the present invention obtains and polysaccharide can be used for preparing health promoting beverage or the protective foods of alleviating physical fatigue; The Semen Cucumidis sativi polypeptide of differing molecular quality can be used for preparation prevention and treatment osteoporosis and promotes medicine or the protective foods of union of fracture, for the utilization of Semen Cucumidis sativi water soluble component, polysaccharide and polypeptide provides a new way;
Four, in the present invention, in Semen Cucumidis sativi, the yield of VITAMIN is 6.6 ‰～10.3 ‰, and in Semen Cucumidis sativi, the yield of fatty oil is 8.1%～26.2%, and in Semen Cucumidis sativi, the yield of polysaccharide is 3.5%～5.8%, and in Semen Cucumidis sativi, the yield of polypeptide is 45.0%～54.8%;
Five, extraction cost of the present invention is compared and has been reduced by 50%～60% with existing methodical extraction cost;
Six, the present invention effectively raises the comprehensive utilization value of Semen Cucumidis sativi medicinal material, for the utilization of Semen Cucumidis sativi water soluble component and residue provides a kind of new way.
The present invention can obtain a kind of method of comprehensive utilization that Semen Cucumidis sativi medicinal material is worth that improves.
Accompanying drawing explanation
Fig. 1 is a kind of method of comprehensive utilization schema that improves Semen Cucumidis sativi medicinal material value of the present invention.
Embodiment
Embodiment one: present embodiment is a kind of method of comprehensive utilization that Semen Cucumidis sativi medicinal material is worth that improves, and comprising:
One, adopt water seaoning to extract the water soluble component in Semen Cucumidis sativi;
Water seaoning described in step 1 comprises: Semen Cucumidis sativi is pulverized, obtained Semen Cucumidis sativi meal; Semen Cucumidis sativi meal is immersed in to 12h～24h in water, refilters, obtain Semen Cucumidis sativi filter residue and filtrate, in filtrate, contain aqueous soluble active constituent; Described aqueous soluble active constituent comprises VITAMIN and organic acid;
The solid-liquid ratio of the Semen Cucumidis sativi meal described in step 1 and water is 1:(5～10);
Two, Semen Cucumidis sativi filter residue step 1 being obtained is ground into powder after drying, then adopts supercritical CO
2extraction process extracts, and obtains Semen Cucumidis sativi fatty oil and residue extract remainder;
Supercritical CO described in step 2
2extraction process, comprises the following steps: at extracting pressure 25MPa～35MPa, 35 ℃～55 ℃ of extraction temperature and CO
2continuous extraction 1h～2h under flow 45L/min～60L/min condition, obtains Semen Cucumidis sativi fatty oil and residue extract remainder;
Three, the residue extract remainder that adopts enzymolysis process to obtain step 2 carries out enzymolysis, deactivation 10min～20min at the temperature of 80 ℃～100 ℃ again, in the enzymolysis solution after deactivation, add distilled water, speed with 5000r/min～10000r/min is carried out centrifugal, get supernatant liquor and filter, obtain the filtrate after enzymolysis; Finally use Semen Cucumidis sativi polysaccharide and Semen Cucumidis sativi polypeptide in the filtrate after the separated enzymolysis of ultrafiltration process;
Enzymolysis solution after deactivation described in step 3 and the volume ratio of distilled water are 1:1;
Enzymolysis process described in step 3, comprises the following steps: in the residue extract remainder obtaining to step 2, add distilled water, according to g/mL meter, the solid-liquid ratio of residue extract remainder and water is 1:6～1:20, and stirring and evenly mixing obtains residue extract remainder/distilled water mixed solution; Regulating the pH of residue extract remainder/distilled water mixed solution is 5.0～8.0, then adds proteolytic enzyme, in temperature, is to stir enzymolysis 3h～8h under 50 ℃～65 ℃ and the stirring velocity condition that is 200r/min～300r/min; The total amount that adds of described proteolytic enzyme is 2500U/g～5500U/g;
Semen Cucumidis sativi polysaccharide in filtrate after the separated enzymolysis of use ultrafiltration process described in step 3 and the method for Semen Cucumidis sativi polypeptide, comprise the following steps:
1. the ultra-filtration membrane ultrafiltration that is 10KDa by the filtrate after enzymolysis through trapped molecular weight, filtrate residual content in ultrafiltration cup after enzymolysis be filtrate cumulative volume after enzymolysis 1/5 time, collect filtered solution, and to supplementing the original volume of distilled water to the filtrate after enzymolysis, ultrafiltration again in ultrafiltration cup;
2. repeating step is 1. 3 times～4 times, stops ultrafiltration, collects each filtered solution and trapped fluid; Step 2. described each filtered solution is the polypeptide liquid of molecular mass <10KDa; The trapped fluid of step described in is 2. the Crude polysaccharides liquid of molecular mass >10KDa;
3. under temperature is the condition of 50 ℃～60 ℃, trapped fluid is evaporated to 1/2～1/3 of trapped fluid initial volume, in the trapped fluid after concentrated, adds ethanol, obtain trapped fluid/ethanolic soln; By the standing 8h～12h of trapped fluid/ethanolic soln, suction filtration, is precipitated, and uses successively dehydrated alcohol, anhydrous diethyl ether and washing with acetone precipitation, and vacuum-drying or lyophilize, obtain Semen Cucumidis sativi polysaccharide powder; In the trapped fluid/ethanolic soln of step described in 3., the volume of ethanol accounts for 60%～95% of trapped fluid/ethanolic soln cumulative volume;
4. the polypeptide liquid of the molecular mass <10KDa 3. step being collected is the ultra-filtration membrane ultrafiltration of 1KDa and 650Da through molecular weight cut-off successively, collect respectively trapped fluid and filtered solution, obtain the polypeptide solution that molecular mass is respectively 1KDa～10KDa, 650Da～1KDa and <650Da, respectively three peptide species solution for vacuum are dried or lyophilize, obtain the Semen Cucumidis sativi polypeptide powder of three kinds of differing molecular quality.
The advantage of present embodiment: one, present embodiment can be extracted simultaneously and utilize the number of chemical compositions such as fatty oil in Semen Cucumidis sativi, polysaccharide, polypeptide, VITAMIN and organic acid, reaches the comprehensive utilization of Semen Cucumidis sativi, has saved resource;
Two, present embodiment extraction process is simple, and extraction conditions is gentle, is conducive to retain the biological activity of its chemical composition, safety and environmental protection;
Three, the Semen Cucumidis sativi water soluble component that present embodiment obtains and polysaccharide can be used for preparing health promoting beverage or the protective foods of alleviating physical fatigue; The Semen Cucumidis sativi polypeptide of differing molecular quality can be used for preparation prevention and treatment osteoporosis and promotes medicine or the protective foods of union of fracture, for the utilization of Semen Cucumidis sativi water soluble component, polysaccharide and polypeptide provides a new way;
Four, in present embodiment, in Semen Cucumidis sativi, the yield of VITAMIN is 6.6 ‰～10.3 ‰, and in Semen Cucumidis sativi, the yield of fatty oil is 8.1%～26.2%, and in Semen Cucumidis sativi, the yield of polysaccharide is 3.5%～5.8%, and in Semen Cucumidis sativi, the yield of polypeptide is 45.0%～54.8%;
Five, the extraction cost of present embodiment is compared and has been reduced by 50%～60% with existing methodical extraction cost;
Six, present embodiment effectively raises the comprehensive utilization value of Semen Cucumidis sativi medicinal material, for the utilization of Semen Cucumidis sativi water soluble component and residue provides a kind of new way.
Present embodiment can obtain a kind of method of comprehensive utilization that Semen Cucumidis sativi medicinal material is worth that improves.
Embodiment two: present embodiment and embodiment one difference are: the proteolytic enzyme described in step 3 be in Sumizyme MP, papoid, neutral protease, composite enzyme for hydrolyzing plant protein, stomach en-and trypsinase a kind of, two or three form in any proportion.Other steps are identical with embodiment one.
Embodiment three: present embodiment and one of embodiment one or two difference are: step 3 4. described ethanol is medical alcohol or dehydrated alcohol.Other steps are identical with embodiment one or two.
Embodiment four: one of present embodiment and embodiment one to three difference is: in step 1, Semen Cucumidis sativi meal is immersed in to 18h～24h in water, refilters, obtain Semen Cucumidis sativi filter residue and filtrate, contain aqueous soluble active constituent in filtrate.Other steps are identical with embodiment one to three.
Embodiment five: one of present embodiment and embodiment one to four difference is: the supercritical CO described in step 2
2extraction process, comprises the following steps: at extracting pressure 30MPa～35MPa, 45 ℃～55 ℃ of extraction temperature and CO
2continuous extraction 1h～2h under flow 50L/min～60L/min condition, obtains Semen Cucumidis sativi fatty oil and residue extract remainder.Other steps are identical with embodiment one to four.
Embodiment six: one of present embodiment and embodiment one to five difference is: adopt the residue extract remainder that enzymolysis process obtains step 2 to carry out enzymolysis in step 3, deactivation 10min at the temperature of 90 ℃～100 ℃ again, in the enzymolysis solution after deactivation, add distilled water, speed with 8000r/min～10000r/min is carried out centrifugal, get supernatant liquor and filter, obtain the filtrate after enzymolysis.Other steps are identical with embodiment one to five.
Embodiment seven: one of present embodiment and embodiment one to six difference is: the enzymolysis process described in step 3, comprise the following steps: in the residue extract remainder obtaining to step 2, add distilled water, according to g/mL, count, the solid-liquid ratio of residue extract remainder and water is 1:10～1:20, stirring and evenly mixing, obtains residue extract remainder/distilled water mixed solution; Regulating the pH of residue extract remainder/distilled water mixed solution is 7.0～8.0, then adds proteolytic enzyme, in temperature, is to stir enzymolysis 5h～8h under 55 ℃～65 ℃ and the stirring velocity condition that is 200r/min～300r/min.Other steps are identical with embodiment one to six.
Embodiment eight: one of present embodiment and embodiment one to seven difference is: the enzymolysis process described in step 3, comprise the following steps: in the residue extract remainder obtaining to step 2, add distilled water, according to g/mL, count, the solid-liquid ratio of residue extract remainder and water is 1:15～1:20, stirring and evenly mixing, obtains residue extract remainder/distilled water mixed solution; Regulating the pH of residue extract remainder/distilled water mixed solution is 5.0～6.0, then adds proteolytic enzyme, in temperature, is to stir enzymolysis 3h～5h under 50 ℃～55 ℃ and the stirring velocity condition that is 200r/min～300r/min.Other steps are identical with embodiment one to seven.
Embodiment nine: one of present embodiment and embodiment one to eight difference is: step 3 is evaporated to 1/2～1/3 of trapped fluid initial volume by trapped fluid in 3. under temperature is the condition of 55 ℃～60 ℃, 10h～12h, suction filtration, be precipitated, use successively dehydrated alcohol, anhydrous diethyl ether and washing with acetone precipitation, vacuum-drying or lyophilize, obtain Semen Cucumidis sativi polysaccharide powder.Other steps are identical with embodiment one to eight.
Embodiment ten: one of present embodiment and embodiment one to nine difference is: in the trapped fluid/ethanolic soln of step 3 described in 3., the volume of ethanol accounts for 60%～95% of trapped fluid/ethanolic soln cumulative volume.Other steps are identical with embodiment one to nine.
Adopt following verification experimental verification beneficial effect of the present invention:
Test one: a kind of method of comprehensive utilization that improves Semen Cucumidis sativi medicinal material value, comprising:
One, adopt water seaoning to extract the water soluble component in Semen Cucumidis sativi;
Water seaoning described in step 1 comprises: Semen Cucumidis sativi is pulverized, obtained Semen Cucumidis sativi meal; Semen Cucumidis sativi meal is immersed in to 24h in water, refilters, obtain Semen Cucumidis sativi filter residue and filtrate, in filtrate, contain aqueous soluble active constituent; Described aqueous soluble active constituent comprises VITAMIN and organic acid;
The solid-liquid ratio of the Semen Cucumidis sativi meal described in step 1 and water is 1:10;
Two, Semen Cucumidis sativi filter residue step 1 being obtained is ground into powder after drying, then adopts supercritical CO
2extraction process extracts, and obtains Semen Cucumidis sativi fatty oil and residue extract remainder;
Supercritical CO described in step 2
2extraction process, comprises the following steps: at extracting pressure 35MPa, 35 ℃ of extraction temperature and CO
2continuous extraction 2h under flow 45L/min condition, obtains Semen Cucumidis sativi fatty oil and residue extract remainder;
Three, the residue extract remainder that adopts enzymolysis process to obtain step 2 carries out enzymolysis, deactivation 10min at the temperature of 100 ℃ again adds distilled water in the enzymolysis solution after deactivation, with the speed of 10000r/min, carries out centrifugal, get supernatant liquor and filter, obtain the filtrate after enzymolysis; Finally use Semen Cucumidis sativi polysaccharide and Semen Cucumidis sativi polypeptide in the filtrate after the separated enzymolysis of ultrafiltration process;
Enzymolysis solution after deactivation described in step 3 and the volume ratio of distilled water are 1:1;
Enzymolysis process described in step 3, comprises the following steps: in the residue extract remainder obtaining to step 2, add distilled water, according to g/mL meter, the solid-liquid ratio of residue extract remainder and water is 1:8, and stirring and evenly mixing obtains residue extract remainder/distilled water mixed solution; Regulating the pH of residue extract remainder/distilled water mixed solution is 8.0, then adds proteolytic enzyme, in temperature, is to stir enzymolysis 6.5h under 50 ℃ and the stirring velocity condition that is 200r/min; Described proteolytic enzyme is Sumizyme MP and neutral protease, and the mass ratio of Sumizyme MP and neutral protease is 3:1; The total amount that adds of described proteolytic enzyme is 4500U/g;
Semen Cucumidis sativi polysaccharide in filtrate after the separated enzymolysis of use ultrafiltration process described in step 3 and the method for Semen Cucumidis sativi polypeptide, comprise the following steps:
1. the ultra-filtration membrane ultrafiltration that is 10KDa by the filtrate after enzymolysis through trapped molecular weight, filtrate residual content in ultrafiltration cup after enzymolysis be filtrate cumulative volume after enzymolysis 1/5 time, collect filtered solution, and to supplementing the original volume of distilled water to the filtrate after enzymolysis, ultrafiltration again in ultrafiltration cup;
2. repeating step is 1. 3 times, stops ultrafiltration, collects each filtered solution and trapped fluid; Step 2. described each filtered solution is the polypeptide liquid of molecular mass <10KDa; The trapped fluid of step described in is 2. the Crude polysaccharides liquid of molecular mass >10KDa;
3. under temperature is the condition of 55 ℃, trapped fluid is evaporated to 1/3 of trapped fluid initial volume, in the trapped fluid after concentrated, adds ethanol, obtain trapped fluid/ethanolic soln; By the standing 12h of trapped fluid/ethanolic soln, suction filtration, is precipitated, and uses successively dehydrated alcohol, anhydrous diethyl ether and washing with acetone precipitation, and vacuum-drying or lyophilize, obtain Semen Cucumidis sativi polysaccharide powder; In the trapped fluid/ethanolic soln of step described in 3., the volume of ethanol accounts for 95% of trapped fluid/ethanolic soln cumulative volume;
4. the polypeptide liquid of the molecular mass <10KDa 3. step being collected is the ultra-filtration membrane ultrafiltration of 1KDa and 650Da through molecular weight cut-off successively, collect respectively trapped fluid and filtered solution, obtain the polypeptide solution that molecular mass is respectively 1KDa～10KDa, 650Da～1KDa and <650Da, respectively three peptide species solution for vacuum are dried or lyophilize, obtain the Semen Cucumidis sativi polypeptide powder of three kinds of differing molecular quality.
The ethanol of this testing sequence three described in is 3. medical alcohol;
In this test, in Semen Cucumidis sativi, the yield of VITAMIN is 9.68 ‰, and in Semen Cucumidis sativi, the yield of fatty oil is 25.65%, and in Semen Cucumidis sativi, the yield of polysaccharide is 5.24%, and in Semen Cucumidis sativi, the yield of polypeptide is 53.6%;
The extraction cost of this test is compared and has been reduced by 55% with existing methodical extraction cost.
Test two: a kind of method of comprehensive utilization that improves Semen Cucumidis sativi medicinal material value, comprising:
One, adopt water seaoning to extract the water soluble component in Semen Cucumidis sativi;
Water seaoning described in step 1 comprises: Semen Cucumidis sativi is pulverized, obtained Semen Cucumidis sativi meal; Semen Cucumidis sativi meal is immersed in to 18h in water, refilters, obtain Semen Cucumidis sativi filter residue and filtrate, in filtrate, contain aqueous soluble active constituent; Described aqueous soluble active constituent comprises VITAMIN and organic acid;
The solid-liquid ratio of the Semen Cucumidis sativi meal described in step 1 and water is 1:8;
Two, Semen Cucumidis sativi filter residue step 1 being obtained is ground into powder after drying, then adopts supercritical CO
2extraction process extracts, and obtains Semen Cucumidis sativi fatty oil and residue extract remainder;
Supercritical CO described in step 2
2extraction process, comprises the following steps: at extracting pressure 30MPa, 45 ℃ of extraction temperature and CO
2continuous extraction 2h under flow 45L/min condition, obtains Semen Cucumidis sativi fatty oil and residue extract remainder;
Three, the residue extract remainder that adopts enzymolysis process to obtain step 2 carries out enzymolysis, deactivation 10min at the temperature of 90 ℃ again adds distilled water in the enzymolysis solution after deactivation, with the speed of 5000r/min, carries out centrifugal, get supernatant liquor and filter, obtain the filtrate after enzymolysis; Finally use Semen Cucumidis sativi polysaccharide and Semen Cucumidis sativi polypeptide in the filtrate after the separated enzymolysis of ultrafiltration process;
Enzymolysis solution after deactivation described in step 3 and the volume ratio of distilled water are 1:1;
Enzymolysis process described in step 3, comprises the following steps: in the residue extract remainder obtaining to step 2, add distilled water, according to g/mL meter, the solid-liquid ratio of residue extract remainder and water is 1:12, and stirring and evenly mixing obtains residue extract remainder/distilled water mixed solution; Regulating the pH of residue extract remainder/distilled water mixed solution is 7.0, then adds proteolytic enzyme, in temperature, is to stir enzymolysis 5h under 50 ℃ and the stirring velocity condition that is 250r/min; Described proteolytic enzyme is composite enzyme for hydrolyzing plant protein and papoid, and the mass ratio of composite enzyme for hydrolyzing plant protein and papoid is 1:1; The total amount that adds of described proteolytic enzyme is 5000U/g;
Semen Cucumidis sativi polysaccharide in filtrate after the separated enzymolysis of use ultrafiltration process described in step 3 and the method for Semen Cucumidis sativi polypeptide, comprise the following steps:
1. the ultra-filtration membrane ultrafiltration that is 10KDa by the filtrate after enzymolysis through trapped molecular weight, filtrate residual content in ultrafiltration cup after enzymolysis be filtrate cumulative volume after enzymolysis 1/5 time, collect filtered solution, and to supplementing the original volume of distilled water to the filtrate after enzymolysis, ultrafiltration again in ultrafiltration cup;
2. repeating step is 1. 4 times, stops ultrafiltration, collects each filtered solution and trapped fluid; Step 2. described each filtered solution is the polypeptide liquid of molecular mass <10KDa; The trapped fluid of step described in is 2. the Crude polysaccharides liquid of molecular mass >10KDa;
3. under temperature is the condition of 60 ℃, trapped fluid is evaporated to 1/2 of trapped fluid initial volume, in the trapped fluid after concentrated, adds ethanol, obtain trapped fluid/ethanolic soln; By the standing 10h of trapped fluid/ethanolic soln, suction filtration, is precipitated, and uses successively dehydrated alcohol, anhydrous diethyl ether and washing with acetone precipitation, and vacuum-drying or lyophilize, obtain Semen Cucumidis sativi polysaccharide powder; In the trapped fluid/ethanolic soln of step described in 3., the volume of ethanol accounts for 85% of trapped fluid/ethanolic soln cumulative volume;
4. the polypeptide liquid of the molecular mass <10KDa 3. step being collected is the ultra-filtration membrane ultrafiltration of 1KDa and 650Da through molecular weight cut-off successively, collect respectively trapped fluid and filtered solution, obtain the polypeptide solution that molecular mass is respectively 1KDa～10KDa, 650Da～1KDa and <650Da, respectively three peptide species solution for vacuum are dried or lyophilize, obtain the Semen Cucumidis sativi polypeptide powder of three kinds of differing molecular quality.
The ethanol of this testing sequence three described in is 3. dehydrated alcohol;
In this test, in Semen Cucumidis sativi, the yield of VITAMIN is 8.54 ‰, and in Semen Cucumidis sativi, the yield of fatty oil is 20.38%, and in Semen Cucumidis sativi, the yield of polysaccharide is 5.76%, and in Semen Cucumidis sativi, the yield of polypeptide is 49.72%;
The extraction cost of this test is compared and has been reduced by 58% with existing methodical extraction cost.
Test three: a kind of method of comprehensive utilization that improves Semen Cucumidis sativi medicinal material value, comprising:
One, adopt water seaoning to extract the water soluble component in Semen Cucumidis sativi;
Water seaoning described in step 1 comprises: Semen Cucumidis sativi is pulverized, obtained Semen Cucumidis sativi meal; Semen Cucumidis sativi meal is immersed in to 12h in water, refilters, obtain Semen Cucumidis sativi filter residue and filtrate, in filtrate, contain aqueous soluble active constituent; Described aqueous soluble active constituent comprises VITAMIN and organic acid;
The solid-liquid ratio of the Semen Cucumidis sativi meal described in step 1 and water is 1:6;
Two, Semen Cucumidis sativi filter residue step 1 being obtained is ground into powder after drying, then adopts supercritical CO
2extraction process extracts, and obtains Semen Cucumidis sativi fatty oil and residue extract remainder;
Supercritical CO described in step 2
2extraction process, comprises the following steps: at extracting pressure 25MPa, 55 ℃ of extraction temperature and CO
2continuous extraction 2h under flow 65L/min condition, obtains Semen Cucumidis sativi fatty oil and residue extract remainder;
Three, the residue extract remainder that adopts enzymolysis process to obtain step 2 carries out enzymolysis, deactivation 10min at the temperature of 85 ℃ again adds distilled water in the enzymolysis solution after deactivation, with the speed of 8000r/min, carries out centrifugal, get supernatant liquor and filter, obtain the filtrate after enzymolysis; Finally use Semen Cucumidis sativi polysaccharide and Semen Cucumidis sativi polypeptide in the filtrate after the separated enzymolysis of ultrafiltration process;
Enzymolysis solution after deactivation described in step 3 and the volume ratio of distilled water are 1:1;
Enzymolysis process described in step 3, comprises the following steps: in the residue extract remainder obtaining to step 2, add distilled water, according to g/mL meter, the solid-liquid ratio of residue extract remainder and water is 1:16, and stirring and evenly mixing obtains residue extract remainder/distilled water mixed solution; Regulating the pH of residue extract remainder/distilled water mixed solution is 7.5, then adds proteolytic enzyme, in temperature, is to stir enzymolysis 7h under 50 ℃ and the stirring velocity condition that is 300r/min; Described proteolytic enzyme is papoid and neutral protease, and the mass ratio of papoid and neutral protease is 2:1; The total amount that adds of described proteolytic enzyme is 5500U/g;
Semen Cucumidis sativi polysaccharide in filtrate after the separated enzymolysis of use ultrafiltration process described in step 3 and the method for Semen Cucumidis sativi polypeptide, comprise the following steps:
1. the ultra-filtration membrane ultrafiltration that is 10KDa by the filtrate after enzymolysis through trapped molecular weight, filtrate residual content in ultrafiltration cup after enzymolysis be filtrate cumulative volume after enzymolysis 1/5 time, collect filtered solution, and to supplementing the original volume of distilled water to the filtrate after enzymolysis, ultrafiltration again in ultrafiltration cup;
2. repeating step is 1. 3 times, stops ultrafiltration, collects each filtered solution and trapped fluid; Step 2. described each filtered solution is the polypeptide liquid of molecular mass <10KDa; The trapped fluid of step described in is 2. the Crude polysaccharides liquid of molecular mass >10KDa;
3. under temperature is the condition of 60 ℃, trapped fluid is evaporated to 1/3 of trapped fluid initial volume, in the trapped fluid after concentrated, adds ethanol, obtain trapped fluid/ethanolic soln; By the standing 8h of trapped fluid/ethanolic soln, suction filtration, is precipitated, and uses successively dehydrated alcohol, anhydrous diethyl ether and washing with acetone precipitation, and vacuum-drying or lyophilize, obtain Semen Cucumidis sativi polysaccharide powder; In the trapped fluid/ethanolic soln of step described in 3., the volume of ethanol accounts for 80% of trapped fluid/ethanolic soln cumulative volume;
4. the polypeptide liquid of the molecular mass <10KDa 3. step being collected is the ultra-filtration membrane ultrafiltration of 1KDa and 650Da through molecular weight cut-off successively, collect respectively trapped fluid and filtered solution, obtain the polypeptide solution that molecular mass is respectively 1KDa～10KDa, 650Da～1KDa and <650Da, respectively three peptide species solution for vacuum are dried or lyophilize, obtain the Semen Cucumidis sativi polypeptide powder of three kinds of differing molecular quality.
The ethanol of this testing sequence three described in is 3. medical alcohol;
In this test, in Semen Cucumidis sativi, the yield of VITAMIN is 7.73 ‰, and in Semen Cucumidis sativi, the yield of fatty oil is 9.72%, and in Semen Cucumidis sativi, the yield of polysaccharide is 4.75%, and in Semen Cucumidis sativi, the yield of polypeptide is 52.38%;
The extraction cost of this test is compared and has been reduced by 56% with existing methodical extraction cost.
Claims (10)
1. improve the method for comprehensive utilization that Semen Cucumidis sativi medicinal material is worth, it is characterized in that a kind of method of comprehensive utilization that Semen Cucumidis sativi medicinal material is worth that improves, comprising:
One, adopt water seaoning to extract the water soluble component in Semen Cucumidis sativi;
Water seaoning described in step 1 comprises: Semen Cucumidis sativi is pulverized, obtained Semen Cucumidis sativi meal; Semen Cucumidis sativi meal is immersed in to 12h～24h in water, refilters, obtain Semen Cucumidis sativi filter residue and filtrate, in filtrate, contain aqueous soluble active constituent; Described aqueous soluble active constituent comprises VITAMIN and organic acid;
The solid-liquid ratio of the Semen Cucumidis sativi meal described in step 1 and water is 1:(5～10);
Two, Semen Cucumidis sativi filter residue step 1 being obtained is ground into powder after drying, then adopts supercritical CO
2extraction process extracts, and obtains Semen Cucumidis sativi fatty oil and residue extract remainder;
Supercritical CO described in step 2
2extraction process, comprises the following steps: at extracting pressure 25MPa～35MPa, 35 ℃～55 ℃ of extraction temperature and CO
2continuous extraction 1h～2h under flow 45L/min～60L/min condition, obtains Semen Cucumidis sativi fatty oil and residue extract remainder;
Three, the residue extract remainder that adopts enzymolysis process to obtain step 2 carries out enzymolysis, deactivation 10min～20min at the temperature of 80 ℃～100 ℃ again, in the enzymolysis solution after deactivation, add distilled water, speed with 5000r/min～10000r/min is carried out centrifugal, get supernatant liquor and filter, obtain the filtrate after enzymolysis; Finally use Semen Cucumidis sativi polysaccharide and Semen Cucumidis sativi polypeptide in the filtrate after the separated enzymolysis of ultrafiltration process;
Enzymolysis solution after deactivation described in step 3 and the volume ratio of distilled water are 1:1;
Enzymolysis process described in step 3, comprises the following steps: in the residue extract remainder obtaining to step 2, add distilled water, according to g/mL meter, the solid-liquid ratio of residue extract remainder and water is 1:6～1:20, and stirring and evenly mixing obtains residue extract remainder/distilled water mixed solution; Regulating the pH of residue extract remainder/distilled water mixed solution is 5.0～8.0, then adds proteolytic enzyme, in temperature, is to stir enzymolysis 3h～8h under 50 ℃～65 ℃ and the stirring velocity condition that is 200r/min～300r/min; The total amount that adds of described proteolytic enzyme is 2500U/g～5500U/g;
Semen Cucumidis sativi polysaccharide in filtrate after the separated enzymolysis of use ultrafiltration process described in step 3 and the method for Semen Cucumidis sativi polypeptide, comprise the following steps:
1. the ultra-filtration membrane ultrafiltration that is 10KDa by the filtrate after enzymolysis through trapped molecular weight, filtrate residual content in ultrafiltration cup after enzymolysis be filtrate cumulative volume after enzymolysis 1/5 time, collect filtered solution, and to supplementing the original volume of distilled water to the filtrate after enzymolysis, ultrafiltration again in ultrafiltration cup;
2. repeating step is 1. 3 times～4 times, stops ultrafiltration, collects each filtered solution and trapped fluid; Step 2. described each filtered solution is the polypeptide liquid of molecular mass <10KDa; The trapped fluid of step described in is 2. the Crude polysaccharides liquid of molecular mass >10KDa;
3. under temperature is the condition of 50 ℃～60 ℃, trapped fluid is evaporated to 1/2～1/3 of trapped fluid initial volume, in the trapped fluid after concentrated, adds ethanol, obtain trapped fluid/ethanolic soln; By the standing 8h～12h of trapped fluid/ethanolic soln, suction filtration, is precipitated, and uses successively dehydrated alcohol, anhydrous diethyl ether and washing with acetone precipitation, and vacuum-drying or lyophilize, obtain Semen Cucumidis sativi polysaccharide powder; In the trapped fluid/ethanolic soln of step described in 3., the volume of ethanol accounts for 60%～95% of trapped fluid/ethanolic soln cumulative volume;
4. the polypeptide liquid of the molecular mass <10KDa 3. step being collected is the ultra-filtration membrane ultrafiltration of 1KDa and 650Da through molecular weight cut-off successively, collect respectively trapped fluid and filtered solution, obtain the polypeptide solution that molecular mass is respectively 1KDa～10KDa, 650Da～1KDa and <650Da, respectively three peptide species solution for vacuum are dried or lyophilize, obtain the Semen Cucumidis sativi polypeptide powder of three kinds of differing molecular quality.
2. a kind of method of comprehensive utilization that Semen Cucumidis sativi medicinal material is worth that improves according to claim 1, it is characterized in that the proteolytic enzyme described in step 3 be in Sumizyme MP, papoid, neutral protease, composite enzyme for hydrolyzing plant protein, stomach en-and trypsinase a kind of, two or three form in any proportion.
3. a kind of method of comprehensive utilization that Semen Cucumidis sativi medicinal material is worth that improves according to claim 1, is characterized in that the ethanol described in step 3 is 4. medical alcohol or dehydrated alcohol.
4. a kind of method of comprehensive utilization that Semen Cucumidis sativi medicinal material is worth that improves according to claim 1, it is characterized in that, in step 1, Semen Cucumidis sativi meal is immersed in to 18h～24h in water, refilter, obtain Semen Cucumidis sativi filter residue and filtrate, in filtrate, contain aqueous soluble active constituent.
5. a kind of method of comprehensive utilization that Semen Cucumidis sativi medicinal material is worth that improves according to claim 1, is characterized in that the supercritical CO described in step 2
2extraction process, comprises the following steps: at extracting pressure 30MPa～35MPa, 45 ℃～55 ℃ of extraction temperature and CO
2continuous extraction 1h～2h under flow 50L/min～60L/min condition, obtains Semen Cucumidis sativi fatty oil and residue extract remainder.
6. a kind of method of comprehensive utilization that Semen Cucumidis sativi medicinal material is worth that improves according to claim 1, it is characterized in that adopting the residue extract remainder that enzymolysis process obtains step 2 to carry out enzymolysis in step 3, deactivation 10min at the temperature of 90 ℃～100 ℃ again, in the enzymolysis solution after deactivation, add distilled water, speed with 8000r/min～10000r/min is carried out centrifugal, get supernatant liquor and filter, obtain the filtrate after enzymolysis.
7. a kind of method of comprehensive utilization that Semen Cucumidis sativi medicinal material is worth that improves according to claim 1, it is characterized in that the enzymolysis process described in step 3, comprise the following steps: in the residue extract remainder obtaining to step 2, add distilled water, according to g/mL, count, the solid-liquid ratio of residue extract remainder and water is 1:10～1:20, stirring and evenly mixing, obtains residue extract remainder/distilled water mixed solution; Regulating the pH of residue extract remainder/distilled water mixed solution is 7.0～8.0, then adds proteolytic enzyme, in temperature, is to stir enzymolysis 5h～8h under 55 ℃～65 ℃ and the stirring velocity condition that is 200r/min～300r/min.
8. a kind of method of comprehensive utilization that Semen Cucumidis sativi medicinal material is worth that improves according to claim 1, it is characterized in that the enzymolysis process described in step 3, comprise the following steps: in the residue extract remainder obtaining to step 2, add distilled water, according to g/mL, count, the solid-liquid ratio of residue extract remainder and water is 1:15～1:20, stirring and evenly mixing, obtains residue extract remainder/distilled water mixed solution; Regulating the pH of residue extract remainder/distilled water mixed solution is 5.0～6.0, then adds proteolytic enzyme, in temperature, is to stir enzymolysis 3h～5h under 50 ℃～55 ℃ and the stirring velocity condition that is 200r/min～300r/min.
9. a kind of method of comprehensive utilization that Semen Cucumidis sativi medicinal material is worth that improves according to claim 1, it is characterized in that during step 3 3., under temperature is the condition of 55 ℃～60 ℃, trapped fluid to be evaporated to 1/2～1/3 of trapped fluid initial volume, in the trapped fluid after concentrated, add ethanol, obtain trapped fluid/ethanolic soln; By the standing 10h～12h of trapped fluid/ethanolic soln, suction filtration, is precipitated, and uses successively dehydrated alcohol, anhydrous diethyl ether and washing with acetone precipitation, and vacuum-drying or lyophilize, obtain Semen Cucumidis sativi polysaccharide powder.
10. a kind of method of comprehensive utilization that Semen Cucumidis sativi medicinal material is worth that improves according to claim 1, is characterized in that the volume of ethanol in the trapped fluid/ethanolic soln described in step 3 3. accounts for 60%～95% of trapped fluid/ethanolic soln cumulative volume.
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