CN104059774A

CN104059774A - Method of preparing purslane polyunsaturated fatty acids from purslane residues

Info

Publication number: CN104059774A
Application number: CN201410298157.7A
Authority: CN
Inventors: 敬思群; 刘璐萍; 杨飞飞; 童莲花
Original assignee: XINJIANG YUANSEN AGRICULTURAL DEVELOPMENT Co Ltd; Xinjiang University
Current assignee: XINJIANG YUANSEN AGRICULTURAL DEVELOPMENT Co Ltd; Xinjiang University
Priority date: 2014-06-30
Filing date: 2014-06-30
Publication date: 2014-09-24
Anticipated expiration: 2034-06-30
Also published as: CN104059774B

Abstract

The invention discloses a method of preparing purslane polyunsaturated fatty acids from purslane residues. The method comprises the following steps: selecting the purslane residues; raising the extraction pressure of subcritical fluid extraction by 0.8MPa and raising the extraction temperature by 40 DEG C, and extracting for 150 minutes; saponifying and hydrolyzing to prepare a fatty acid mixed before enrichment; heating and stirring urea and anhydrous ethanol for reflux at 60 DEG C in a ratio m/m of urea and fatty acid mixed before enrichment of 1 to 1 and in a ratio m/v of fatty acid mixed before enrichment and ethanol of 1 to 16; after dissolving urea and uniformly mixing with the fatty acid mixed before enrichment, heating and stirring for reflux for 20 minutes; cooling to room temperature; crystallizing for 24 hours at 4 DEG C; filtering to obtain the fatty acid mixed after enrichment. Verified by diabetic rat test, the prepared purslane polyunsaturated fatty acid microcapsule and soft capsule can effectively reduce the levels of blood glucose and insulin and have wide application value and prospect in preparing foods or medicines for preventing and treating diabetes.

Description

A kind of method of preparing purslane polyunsaturated fatty acid from purslane slag

Technical field

The present invention relates to the technical field that polyunsaturated fatty acid extracts preparation.Specifically, the present invention relates to a kind of preparing technical field of preparing purslane polyunsaturated fatty acid from purslane slag.

Background technology

Polyunsaturated fatty acid is a kind of indispensable nutritive substance growing to the mankind or animal, mainly comprise that linolic acid, linolenic acid and timnodonic acid (EPA) also have docosahexenoic acid (DHA) etc., wherein linolic acid, alpha-linolenic acid can not synthesize in human body, can only obtain from absorbing in the middle of food, therefore be called the indispensable fatty acid of human body.It has a lot of effects, and such as reducing blood-fat, hypotensive, antithrombotic, raising immunity of organisms, prevention and adjusting cardiovascular disorder etc., it is widely used in the every field of food, health care etc. with unique nutritive ingredient and pharmaceutical use.Up to the present, the main source of polyunsaturated fatty acid is main mainly with plant, animal and microorganism etc., but at home the exploitation present situation of polyunsaturated fatty acid is also difficult to meet the demand in market.

The advantages such as purslane is drought-resistant with it, disease and insect resistance, vitality are strong are the high and adaptable cash crop of a kind of output in China, are one of wild plants of the integration of drinking and medicinal herbs assert of health ministry.Prior art research shows, purslane oil carries out linoleic acid content in fatty acid component and accounts for fatty acid total amount 33.19%, and linolenic acid content accounts for 31.04%.Therefore from purslane slag, extracting and enriching obtains polyunsaturated fatty acid and prepares the products such as polyunsaturated fatty acid microcapsule, soft capsule to have wide market outlook.

Microcapsule ( microencapsule) be to using natural or synthetic macromolecular material as wall material, by Physical, chemical method or physico-chemical processes, a kind of active substance (core) is wrapped up and forms the minigel with semipermeability or sealing cyst membrane.During micro encapsulation, by the material of embedding, be known as core, the film forming material of embedding core is called as wall material.Wall material is normally formed by natural or synthetic macromolecular material, in micro encapsulation process, need to possess good mobility and solvability, promote the parcel of core to form more stable emulsion system, the method for micro encapsulation has spray-drying process, Inclusion Complexes method, phase separation method and interfacial polymerization etc.Because polyunsaturated fatty acid self exists many unfavorable factors, greatly limited its use value, adopt microcapsulary to carry out capsulation processing, can improve its stability and the operability in functional product thereof, promote the performance of its physiological function.Have no at present about prepare research and the report of purslane polyunsaturated fatty acid microcapsule from purslane slag, from purslane slag, prepare purslane polyunsaturated fatty acid microcapsule and exist many technical bottlenecks to solve.

Soft capsule is a kind of novel form growing up after tablet, injection, can be by oily medicine, drug solution or drug suspension, mashed prod even medical solid powder, particle quantitative pressure injection be encapsulated in capsule, form size, different seal capsule.Compare with other formulation, soft capsule has that bioavailability is high, the uniform and stable property of liquid is good, sealingly secure, content are accurate, aesthetic in appearance, cover the features such as unpleasant odor, be applicable to that taking dose is little, hydrophobic drug, low melting point substance, oil various for health care that oleaginousness is high, to photaesthesia, meet the production of damp and hot unstable, oxidizable and easy volatile medicine, be widely used in pharmacy, food, makeup, household supplies, chemical industry and other field, it is developed, Application Areas is very extensive.Have no at present about prepare research and the report of purslane polyunsaturated fatty acid soft capsule from purslane slag, from purslane slag, prepare purslane polyunsaturated fatty acid soft capsule and exist many technical bottlenecks to solve.

Summary of the invention

For having no in prior art about prepare research and the report of purslane polyunsaturated fatty acid from purslane slag, from purslane slag prepare purslane polyunsaturated fatty acid exist as little in supercritical extracting equipment volume, cost is high, power consumption is large, be not suitable for the defect of large-scale commercial production and polyunsaturated fatty acid efficiently many technical bottlenecks such as purification need to solve.The technical issues that need to address of the present invention are just to overcome the defect of prior art, aim to provide a kind of method of preparing purslane polyunsaturated fatty acid from purslane slag, selecting purslane slag is raw material, by subcritical fluid extraction method, extract purslane oil, and utilize urea clathration technology to carry out separation and purification to the polyunsaturated fatty acid in purslane oil, by adopting subcritical fluid extraction technology and urea clathration technology integrated technique, arrange and mix wall material, vacuum freeze-drying technique is prepared microcapsule, arrange and mix wall material, hot melt process pelleting technique is prepared soft capsule, and there is significant effect of lowering blood sugar through efficacy test proof purslane polyunsaturated fatty acid microcapsule and soft capsule, there is nutritive value and using value widely, effectively overcome the technical bottleneck existing in the efficient purification of current polyunsaturated fatty acid, having real widespread use is worth.

Technical scheme provided by the invention:

The present invention take that to produce the purslane slag that purslane inspissated juice left behind be raw material, by adopting subcritical fluid extraction technology and urea clathration technology integrated technique, selecting purslane slag in purslane processing is that raw material prepares mixed fatty acid after the enrichment that contains high purity polyunsaturated fatty acid, arrange and mix wall material, pass through magnetic agitation, high-speed homogenization, vacuum freeze-drying technique is prepared microcapsule, by hot melt process pelleting technique, prepare soft capsule, effectively overcome the technical bottleneck existing in the efficient purification of current polyunsaturated fatty acid, for the Application and Development research of purslane polyunsaturated fatty acid provides real basis, be with a wide range of applications.

The present invention specifically provides a kind of method of preparing purslane polyunsaturated fatty acid from purslane slag, and product form is to embody with purslane polyunsaturated fatty acid microcapsule and two kinds of product forms of purslane polyunsaturated fatty acid soft capsule.

Concrete, the invention provides a kind of method of preparing purslane polyunsaturated fatty acid microcapsule from purslane slag, % is according to weight percent meter, and concrete grammar step is as follows:

(1) preparation of purslane slag: take fresh purslane, Drinking Water is main raw material, fresh purslane is selected, clean, chopping, by weight 1:3 ratio, soaks 12h, and infusion 6h at 80 ℃, filters and obtains purslane slag.

(2) adopt the pre-treatment that purslane slag is raw material: the purslane slag that the purslane slag in the processing of employing purslane or above-mentioned steps provide is raw material, through 70 ℃ of dry, pulverize, sieve 60 orders, weighing, fillers.

(3) subcritical Trimethylmethane abstraction technique extracts purslane oil: carry out subcritical fluid extraction technique, impose a condition, extracting pressure 0.8MPa, 40 ℃ of extraction temperature, extraction 150min, by the purslane oil extractum through extraction preparation, wait extract the purslane oil extractum that after three times, merging obtains at every turn, after the centrifugal 15min of 4000r/min, filtration, prepare finished product purslane oil.

(4) saponification is prepared the front mixed fatty acid of enrichment: according to g/mL, count, purslane oil is mixed by 1:20 with 0.5mol/L KOH-ethanolic soln, the purslane oil that soon prepared by above-mentioned steps (3) adds the ethanolic soln of KOH, then be put in stirred in water bath, the backflow 120min of 75 ℃, obtain saponification liquor; Add with the ice distilled water of 0.5 mol/L KOH-ethanolic soln equivalent cooling rapidly, then use the petroleum ether extraction unsaponifiables with 0.5 mol/L KOH-ethanolic soln equivalent, through extraction repeatedly, the water gaging dilution that doubles of water intaking layer, slowly adds 10% HCl, is adjusted to pH=2 ~ 3, now can obviously see oil reservoir and water layer, use immediately the petroleum ether extraction mixed fatty acid with 0.5mol/LKOH-ethanolic soln equivalent, extraction repeatedly, obtains petroleum ether layer by separating funnel; Petroleum ether layer is washed till neutrality by 5% NaCl solution water, then with rotary evaporation after anhydrous sodium sulfate drying, filtration, obtains the front mixed fatty acid of enrichment .

(5) adopt urea adduct method enriching polyunsaturated fatty acid: according to g/g/mL, count, by the mixed fatty acid before enrichment, urea and dehydrated alcohol are according to 1:1:16, with the mixed fatty acid before urea/enrichment, press m/m, be 1, mixed fatty acid/ethanol before enrichment is 1:16 by m/v, by urea and dehydrated alcohol, heated and stirred at 60 ℃ refluxes, until urea all dissolves, pour into rapidly in the mixed fatty acid filling before enrichment prepared by above-mentioned steps (4), after mixing, heated and stirred backflow 20min at 60 ℃, be cooled to room temperature, be placed in after 4 ℃ of crystallization 24h, by the rapid suction filtration of crystal clathrate, obtain crystal clathrate and filtrate two portions, wherein in crystal clathrate, contain most saturated fatty acids and oleic acid, by inclusion lipid acid not in filtrate, be about to highly purified polyunsaturated fatty acid through extraction, after being removed to ethanol, filtrate rotary evaporation obtains solid particulate, add 30% water, HCL is adjusted to pH5 ~ 6, then adds with the sherwood oil of dehydrated alcohol equivalent and extract, with distillation washing ether layer to urea detection paper without urea till, use anhydrous sodium sulfate drying 4h, mixed fatty acid after the enrichment of the polyunsaturated fatty acid that can obtain containing higher degree,

(6) preparation of polyunsaturated fatty acid microcapsule: select gum arabic, beta-cyclodextrin and modified starch as wall material, the mixed fatty acid after enrichment of take is core, with wall material and core according to weight ratio 3:1 proportioning, after being mixed according to weight ratio 1:1:2, the gum arabic taking, beta-cyclodextrin and modified starch stir 30min under 60 ℃ of conditions, then through stirring, it is fully dissolved, and placement room temperature is cooling, obtains water; Take mixed fatty acid after 25% enrichment prepared by above-mentioned steps (5), and add 2% tween 80 to carry out stirring and dissolving under 60 ℃ of conditions, obtain oil phase; Oil phase is slowly joined in water and continues to stir under 60 ℃ of conditions, it is fully dissolved, then use under high speed homogenization 10000r/min condition and disperse 3min, make oil droplet uniformly emulsify in wall material solution, be milk sap state, finally be placed in the dry 12h of vacuum freezing drying oven of-55 ℃, collect product, prepare purslane polyunsaturated fatty acid microcapsule.

Meanwhile, the present invention specifically provides a kind of method of preparing purslane polyunsaturated fatty acid soft capsule from purslane slag, and % is according to weight percent meter, and concrete grammar step is as follows:

(2) adopt the pre-treatment that purslane slag is raw material: purslane slag prepared by the purslane slag in the processing of employing purslane or above-mentioned steps is raw material, through 70 ℃ of dry, pulverize, sieve 60 orders, weighing, fillers.

(5) adopt urea adduct method enriching polyunsaturated fatty acid: according to g/g/mL, count, by the mixed fatty acid before enrichment, urea and dehydrated alcohol are according to 1:1:16, with the mixed fatty acid before urea/enrichment, press m/m, be 1, mixed fatty acid/ethanol before enrichment is 1:16 by m/v, by urea and dehydrated alcohol, heated and stirred at 60 ℃ refluxes, until urea all dissolves, pour into rapidly in the mixed fatty acid filling before enrichment prepared by above-mentioned steps (4), after mixing, heated and stirred backflow 20min at 60 ℃, be cooled to room temperature, be placed in after 4 ℃ of crystallization 24h, by the rapid suction filtration of crystal clathrate, obtain crystal clathrate and filtrate two portions, wherein in crystal clathrate, contain most saturated fatty acids and oleic acid, by inclusion lipid acid not in filtrate, be about to highly purified polyunsaturated fatty acid through extraction, after being removed to ethanol, filtrate rotary evaporation obtains solid particulate, add 30% water, HCL is adjusted to pH5 ~ 6, then adds with the sherwood oil of dehydrated alcohol equivalent and extract, with distillation washing ether layer to urea detection paper without urea till, use anhydrous sodium sulfate drying 4h, mixed fatty acid after the enrichment of the polyunsaturated fatty acid that can obtain containing higher degree.

(6) preparation of polyunsaturated fatty acid soft capsule: select gum arabic, beta-cyclodextrin and modified starch as wall material, the mixed fatty acid of take after enrichment is core, with wall material and core according to weight ratio 3:1 proportioning, after being mixed according to weight ratio 1:1:2, the gum arabic taking, beta-cyclodextrin and modified starch stir 30min under 60 ℃ of conditions, then through stirring, it is fully dissolved, and placement room temperature is cooling, obtains water; Take the mixed fatty acid after 25% enrichment prepared by above-mentioned steps (5), and add 2% tween 80 to carry out stirring and dissolving under 60 ℃ of conditions, obtain oil phase; Oil phase is slowly joined in water and continues to stir under 60 ℃ of conditions, it is fully dissolved, then use under high speed homogenization 10000r/min condition and disperse 3min, make oil droplet uniformly emulsify in wall material solution, be milk sap state; In glue pot, first add glycerine (15%), pure water (40%), add gelatin (45%) after stirring and be heated to 75 ℃, stirring and dissolving 60min dissolves gelatin completely, vacuumizes, and removes bubble, 60 ℃ of heat preservation for standby use; Glue is packed in capsule machine processed and carries out pelleting with the content mixing, seasoning after capsule and pill sizing, temperature is controlled at 45 ℃～50 ℃, prepares purslane polyunsaturated fatty acid soft capsule.

In technical scheme provided by the invention, take into full account the polyunsaturated fatty acid in purslane oil, mainly refer to that linolenic and linoleic all exists with the form of glyceryl ester, and glyceryl ester is sterically hindered larger, be difficult to direct inclusion separated, for this reason in adopting urea adduct method enrichment purslane oil before polyunsaturated fatty acid, first glycerin fatty acid ester is carried out to saponification and make mixed fatty acid before enrichment; The present invention by saponification react preparation enrichment before mixed fatty acid, the optimum process condition of saponification is: KOH-alcohol concn is 0.5 mol/L, saponification temperature is 75 ℃, saponification time is 120min, mixed fatty acid yield is 57.36%, polyunsaturated fatty acid before enrichment in mixed fatty acid accounts for 68.12%, is mainly linolic acid, linolenic acid.

The present invention is by the above-mentioned preparation method who provides, by adopting subcritical fluid extraction method, extract purslane oil, and utilize urea clathration technology to carry out separation and purification to the polyunsaturated fatty acid in purslane oil, mixed fatty acid prepare polyunsaturated fatty acid microcapsule and soft capsule after the enrichment that obtains containing high purity polyunsaturated fatty acid, the purslane polyunsaturated fatty acid microcapsule product color of preparation is milk yellow, without special odor, powder is evenly fine and smooth; Surface oil length is 17.3%, and embedding rate is 82.7%; Microcapsule water ratio is 3.80%, meets the requirement of microcapsule moisture content; Median size is 306.6nm.The purslane polyunsaturated fatty acid soft capsule product colour of preparation is faint yellow, and without special odor, granularity is moderate.

Further, the invention provides the effect of lowering blood sugar experiment of purslane polyunsaturated fatty acid microcapsule and soft capsule: the dosage of setting purslane polyunsaturated fatty acid microcapsule and soft capsule is consistent, the impact of research various dose on the function of blood sugar reduction of diabetes rat.By the experiment of rat function of blood sugar reduction, confirm: the hypoglycemic while falls in purslane polyunsaturated fatty acid microcapsule and soft capsule, also can the raise content of diabetes rat serum insulin, relatively has significant difference (P<0.01) with model group rat.Hence one can see that, and purslane polyunsaturated fatty acid microcapsule and soft capsule can effectively reduce blood sugar and the insulin level of diabetes rat, for prevention and the treatment of diabetes, have positive effect.Visible, the present invention is tested and is confirmed that purslane polyunsaturated fatty acid microcapsule and the soft capsule of preparation can effectively reduce blood sugar and insulin level by diabetes rat, prevention and treatment for diabetes have positive effect, show that purslane polyunsaturated fatty acid microcapsule provided by the invention and purslane polyunsaturated fatty acid soft capsule are with a wide range of applications and prospect in the food of preparation prevention and treatment diabetes or medicine.

By implementing the concrete summary of the invention of the present invention, can reach following beneficial effect:

(1) first the present invention adopts subcritical Trimethylmethane abstraction technique to extract purslane oil, investigated the impact on purslane oil extraction yield of single factors such as extracting pressure, extraction temperature, extraction time, according to single factor design orthogonal experiment to determine best processing parameter.Result shows: the optimum process condition of the purslane oil of subcritical fluid extraction: extracting pressure 0.8MPa, 40 ℃ of extraction temperature, extraction time 150min, extraction yield is 2.91%, compares and has improved 1.2% with ultrasonic assistance enzymolysis-Soxhlet extraction coupling method of the common employing in this area.

(2) the present invention prepares purslane oil by adopting the ultrasonic assistance enzymolysis-Soxhlet of subcritical fluid extraction method and the common employing in this area to extract two kinds of methods of coupling method, by mensuration and the lipid acid GC-MS of purslane oil physical and chemical index are analyzed to the quality that has compared two kinds of oil extracting methods.Result shows: adopt the content of polyunsaturated fatty acid of traditional ultrasonic assistance enzymolysis-Soxhlet coupling method gained purslane oil to reach 56.39%, the content of polyunsaturated fatty acid of subcritical fluid extraction purslane oil reaches 64.23%, than front method, has improved 7.84%.It is low that the purslane oleic acid value of subcritical fluid extraction and peroxide value are extracted coupling method than ultrasonic enzymolysis-Soxhlet, and quality will be got well.

(3) the present invention adopts saponification to prepare mixed fatty acid before enrichment to purslane oil, the mixed fatty acid yield of take before enrichment has been investigated KOH-ethanol, saponification time, the impact of saponification temperature three factors on the saponification reaction of purslane oil as index, according to single factor design orthogonal experiment to determine optimal processing parameter.The optimum process condition of saponification is: KOH-alcohol concn is 0.5 mol/L, and saponification temperature is 75 ℃, and saponification time is 120min, amplifies confirmatory experiment, and obtaining the front mixed fatty acid yield of enrichment is 57.36%.

(4) the present invention adopts urea adduct method enriching polyunsaturated fatty acid, iodine number and the rate of recovery of mixed fatty acid after enrichment of take is to judge index, the front mixed fatty acid (m/m) of urea/enrichment, the front mixed fatty acid/ethanol (m/v) of enrichment, return time and the impact of inclusion time on urea clathration reaction have been investigated, by single factor and Orthogonal experiment results, determine urea clathration optimal conditions, utilize GC-MS to analyze the product fatty acid component after purifying; Result shows: urea clathration optimised process is: before urea/enrichment, mixed fatty acid (m/m) is 1, before enrichment, mixed fatty acid/ethanol (m/v) is 1:16, return time 20min, inclusion time 24h, after the enrichment obtaining under this technique, mixed fatty acid is compared with the lipid acid of purslane oil before inclusion not, and content of polyunsaturated fatty acid brings up to 94.56% by 64.23%.

(5) by adopting the method for preparing purslane polyunsaturated fatty acid from purslane slag provided by the invention, the purslane polyunsaturated fatty acid microcapsule product color of preparation is milk yellow, and without special odor, powder is evenly fine and smooth; Surface oil length is 17.3%, and embedding rate is 82.7%; Microcapsule water ratio is 3.80%, meets the requirement of microcapsule moisture content; Median size is 306.6nm.The purslane polyunsaturated fatty acid soft capsule product colour of preparation is faint yellow, and without special odor, granularity is moderate.The present invention is tested and is confirmed that purslane polyunsaturated fatty acid microcapsule and the soft capsule of preparation can effectively reduce blood sugar and insulin level by diabetes rat, prevention and treatment for diabetes have positive effect, in the food of preparation prevention and treatment diabetes or medicine, are with a wide range of applications and prospect.

Accompanying drawing explanation

Fig. 1 prepares the process flow sheet of purslane polyunsaturated fatty acid capsule from purslane slag.

Fig. 2 is shown as the affect figure of extracting pressure on extraction yield.

Fig. 3 is shown as the affect figure of extraction temperature on extraction yield.

Fig. 4 is shown as the affect figure of extraction time on extraction yield.

Fig. 5 is shown as the affect figure of extraction times on purslane oil extraction yield.

Fig. 6 be shown as KOH-alcohol concn on enrichment before mixed fatty acid yield affect figure.

Fig. 7 be shown as saponification temperature on enrichment before mixed fatty acid yield affect figure.

Fig. 8 be shown as saponification time on enrichment before mixed fatty acid yield affect figure.

Fig. 9 is shown as the affect figure of the front mixed fatty acid (m/m) of urea/enrichment on mixed fatty acid urea clathration after enrichment.

Figure 10 is shown as the affect figure of the front mixed fatty acid/ethanol (m/v) of enrichment on mixed fatty acid urea clathration after enrichment.

Figure 11 be shown as the inclusion time on enrichment after mixed fatty acid urea clathration affect figure.

Embodiment

Below, for embodiment, the present invention is described, still, the present invention is not limited to following embodiment, and the % substantially relating in the present invention is weight percent meter without specializing.

All raw and auxiliary materials of selecting in the present invention, selected all reagent and instrument are all well known selecting, but do not limit enforcement of the present invention, other reagent more well known in the art and equipment are all applicable to the enforcement of the following embodiment of the present invention.

embodiment mono-: the preparation of purslane polyunsaturated fatty acid microcapsule

%according to weight percent meter, the concrete grammar step of preparing purslane polyunsaturated fatty acid microcapsule from purslane slag is as follows:

The purslane polyunsaturated fatty acid microcapsule product color of preparing through above-mentioned preparation method is milk yellow, and without special odor, powder is evenly fine and smooth; Surface oil length is 17.3%, and embedding rate is 82.7%; Microcapsule water ratio is 3.80%, meets the requirement of microcapsule moisture content; Median size is 306.6nm.

embodiment bis-: the preparation of purslane polyunsaturated fatty acid soft capsule

%according to weight percent meter, the concrete grammar step of preparing purslane polyunsaturated fatty acid soft capsule from purslane slag is as follows:

By the purslane polyunsaturated fatty acid soft capsule of above-mentioned preparation, after enrichment mixed fatty acid lipid acid with respect to enrichment before the overall resistance of oxidation of mixed fatty acid, purslane oil remarkable, the purslane polyunsaturated fatty acid soft capsule product colour of preparation is faint yellow, without special odor, granularity is moderate.

embodiment tri-: the preparation of purslane polyunsaturated fatty acid microcapsule and soft capsule

By weight percentage, the concrete grammar step of purslane polyunsaturated fatty acid microcapsule and soft capsule is as follows for %:

(1) adopt the pre-treatment that purslane slag is raw material: take fresh purslane, Drinking Water is main raw material, fresh purslane is selected, clean, chopping, by weight 1:3 ratio, soak 12h, infusion 6h at 80 ℃, filter and obtain purslane slag, or to adopt the purslane slag in purslane processing be raw material, purslane slag 70 ℃ of dry, pulverize, sieve 60 orders, weighing, fillers.

(2) subcritical Trimethylmethane abstraction technique extracts purslane oil: adopt subcritical fluid extraction technique, impose a condition: extracting pressure 0.8MPa, 40 ℃ of extraction temperature, extraction 150min, by the purslane oil extractum through extraction preparation, wait extract the purslane oil extractum that after three times, merging obtains at every turn, after the centrifugal 15min of 4000r/min, filtration, prepare finished product purslane oil.

(3) saponification is prepared the front mixed fatty acid of enrichment: the purslane of being prepared by above-mentioned steps (2) oil that takes 1g, the ethanolic soln that adds the KOH of 20mL 0.5 mol/L concentration, then be put in stirred in water bath, the backflow 120min of 75 ℃, obtain saponification liquor; Add with the ice distilled water of 0.5mol/LKOH-ethanolic soln equivalent cooling rapidly, then use the 30ml petroleum ether extraction unsaponifiables with 0.5mol/LKOH-ethanolic soln equivalent, through extraction repeatedly, the water gaging dilution that doubles of water intaking layer, slowly adds 10% HCl, is adjusted to pH=2 ~ 3, now can obviously see oil reservoir and water layer, use immediately the petroleum ether extraction mixed fatty acid with the 20ml of 0.5mol/LKOH-ethanolic soln equivalent, extraction repeatedly, obtains petroleum ether layer by separating funnel; Petroleum ether layer is washed till neutrality by 5% NaCl solution water, then with rotary evaporation after anhydrous sodium sulfate drying, filtration, obtains the front mixed fatty acid of enrichment .

(4) adopt urea adduct method enriching polyunsaturated fatty acid: urea and 16mL dehydrated alcohol heated and stirred at 60 ℃ of getting 1g reflux, until urea all dissolves, pour into rapidly before the enrichment that fills 1g prepared by above-mentioned steps (3) in mixed fatty acid, after mixing, heated and stirred backflow 20min at 60 ℃, be cooled to room temperature, be placed in after 4 ℃ of crystallization 24h, by the rapid suction filtration of crystal clathrate, obtain crystal clathrate and filtrate two portions, wherein in crystal clathrate, contain most saturated fatty acids and oleic acid; By inclusion lipid acid not in filtrate, be about to highly purified polyunsaturated fatty acid through extraction, after being removed to ethanol, filtrate rotary evaporation obtains solid particulate, add 30% 5mL water, dense HCL is adjusted to pH5 ~ 6, then adds with the sherwood oil of dehydrated alcohol equivalent and extract, with distillation washing ether layer to urea detection paper without urea till, use anhydrous sodium sulfate drying 4h, mixed fatty acid after the enrichment of the polyunsaturated fatty acid that can obtain containing higher degree.

(5) preparation of polyunsaturated fatty acid microcapsule: select gum arabic, beta-cyclodextrin and modified starch as wall material, the mixed fatty acid before enrichment of take is core, with wall material and core according to weight ratio 3:1 proportioning; After the gum arabic taking, beta-cyclodextrin and modified starch are mixed according to weight ratio 1:1:2, under 60 ℃ of conditions, stir 30min, then, through stirring, it is fully dissolved, placement room temperature is cooling, obtains water; Take the front mixed fatty acid of 25% enrichment prepared by above-mentioned steps (4), and add 2% tween 80 to carry out stirring and dissolving under 60 ℃ of conditions, obtain oil phase; Oil phase is slowly joined in water and continues to stir under 60 ℃ of conditions, it is fully dissolved, then use under high speed homogenization 10000r/min condition and disperse 3min, make oil droplet uniformly emulsify in wall material solution, be milk sap state, finally be placed in the dry 12h of vacuum freezing drying oven of-55 ℃, collect product, prepare purslane polyunsaturated fatty acid microcapsule.

(6) preparation of polyunsaturated fatty acid soft capsule: select gum arabic, beta-cyclodextrin and modified starch as wall material, the mixed fatty acid of take after enrichment is core, with wall material and core according to weight ratio 3:1 proportioning, after being mixed according to weight ratio 1:1:2, the gum arabic taking, beta-cyclodextrin and modified starch stir 30min under 60 ℃ of conditions, then through stirring, it is fully dissolved, and placement room temperature is cooling, obtains water; Take the mixed fatty acid after 25% enrichment prepared by above-mentioned steps (4), and add 2% tween 80 to carry out stirring and dissolving under 60 ℃ of conditions, obtain oil phase; Oil phase is slowly joined in water and continues to stir under 60 ℃ of conditions, it is fully dissolved, then use under high speed homogenization 10000r/min condition and disperse 3min, make oil droplet uniformly emulsify in wall material solution, be milk sap state; In glue pot, first add glycerine (15%), pure water (40%), add gelatin (45%) after stirring and be heated to 75 ℃, stirring and dissolving 60min dissolves gelatin completely, vacuumizes, and removes bubble, 60 ℃ of heat preservation for standby use; Glue is packed in capsule machine processed and carries out pelleting with the content mixing, seasoning after capsule and pill sizing, temperature is controlled at 45 ℃～50 ℃, prepares purslane polyunsaturated fatty acid soft capsule.

embodiment tetra-: subcritical fluid extraction purslane oil

1.1 subcritical fluid extraction purslanes oil briefs description of the process: first by purslane slag drying, pulverize, sieve, take a certain amount of being sub-packed in 200 order Nylon Bags, tighten sack, drop in batch extractor, take out batch extractor negative pressure to-0.08MPa.Solvent is imported to batch extractor by solvent tank, and because batch extractor volume is larger, a small amount of error of each extraction solvent amount is less on experimental result impact, till therefore being observed and made solvent flood the about 5cm of material by visor.Open into hot water valve and hot water pump batch extractor is carried out to circulating-heating, when temperature rises to certain temperature, oil in purslane is fully dissolved, after extraction certain hour, with force (forcing) pump, extraction liquid is imported to evaporating pot, by heating, solvent is constantly gasified, with system compresses machine, solvent vapo(u)r is recycled to solvent tank by condenser and continues to use, open evaporating pot delivery valve, the purslane oil extractum soaking with clean container access first.Until extraction, repeatedly merge afterwards the purslane oil extractum at every turn obtaining, through centrifugal, filter after both finished product purslane oil.

1.2 purslane oil percentage extraction method of calculation: percentage extraction=

In formula: m ₁for the quality (g) of purslane, m ₂quality (g) for the purslane oil that extracts.

1.3 experiment of single factor

1.3.1 the impact of extracting pressure on purslane oil extraction yield

Get and pulverize and sieve sample 4kg after (60 order), 30 ℃ of extraction temperature, extraction time 120min, investigate in 0.7MPa, 0.8MPa, 0.9MPa, 1.0MPa, the impact of the different extracting pressure of 1.1MPa on purslane oil extraction effect.

From accompanying drawing 2, purslane oil extraction yield increases along with the rising of extracting pressure, and when pressure, purslane oil extraction rate reached is to maximum value 2.29% during at 0.8MPa, and after surpassing 0.8 MPa, extraction yield trend is stablized.Within the scope of subcritical region, the increase of pressure increases the density of fluid solvent, and dissolving power also increases thereupon.And along with the increase of fluid solvent pressure, the interaction in solute molecule also increases, so the solvability of purslane oil is also increase trend, so extraction yield increases.When pressure is during higher than 0.8MPa, extracting pressure is not remarkable on the impact of extraction yield, shows that now the increase of pressure is less on the solvability impact of purslane oil.Therefore, extracting pressure of the present invention selects 0.8MPa to be advisable.

1.3.2 the impact of extraction temperature on purslane oil extraction yield:

Get and pulverize and sieve sample 4kg after (60 order), extracting pressure is 0.8MPa, and extraction time 120min investigates the impact of 25 ℃, 30 ℃, 35 ℃, 40 ℃, 45 ℃ different extraction temperature on the oily extraction effect of purslane.

From accompanying drawing 3, in extraction for the previous period, the extraction yield of purslane oil is along with temperature raises and increases.When temperature rises to 35 ℃ from 25 ℃, extraction yield significantly increases, and rises to 2.57% from 1.87%, obtains maximum extracted rate in the time of 35 ℃.The rising of this explanation temperature can make the mass transfer coefficient of fluid increase, and viscosity degradation, is conducive to extraction.But when temperature continues to rise, extraction yield is on a declining curve, this is due to the density that can reduce fluid that increases of temperature, and solvent has part vaporization trend.Not only can not improve percentage extraction, increase on the contrary System risk, also be unfavorable for bioactive reservation in product, energy consumption consumption also increases simultaneously, therefore selects 35 ℃ as the optimum temperuture of extraction.

1.3.3 the impact of extraction time on purslane oil extraction yield

Get and pulverize and sieve sample 4kg after (60 order), extracting pressure is 0.8MPa, and 35 ℃ of extraction temperature are investigated 60min, 90min, 120min, 150min, the impact of the different extraction times of 180min on purslane oil extraction effect.

Grease reaches dissolution equilibrium in subcritical fluids needs the regular hour.From accompanying drawing 4, with extraction time, extend, extraction yield presents rising trend.When extraction time is 120 min, reach maximum value 2.6%, afterwards along with time lengthening, extraction yield declines, and likely the prolongation of extraction time causes impurity stripping quantity to increase, and makes purslane oil extraction yield present downtrending.Extend extraction time, extraction yield has to decline and tend to slightly to be stablized again.This has also illustrated that extraction time is not that the longer the better, extends simply extraction time, and energy consumption raises, and extraction efficiency declines.

1.3.4 the impact of extraction times on purslane oil extraction yield

Get and pulverize and sieve sample 4kg after (60 order), extracting pressure is 0.8MPa, 35 ℃ of extraction temperature, and extraction time 40min, investigates the impact of extraction times on purslane oil extraction effect.

Different from supercritical fluid extraction: supercritical extraction can be realized continuous extraction, parsing, and subcritical abstraction due to extraction agent in resolving and extract velocity of separation slower, therefore can only realize intermittent type lixiviate.From accompanying drawing 5, extraction times is comparatively remarkable on the impact of subcritical abstraction, and this is because fluid is very strong to the solvability of grease.When extraction times to 3 time, purslane oil extraction yield has reached 2.73%, and along with the increase of extraction times, grease is almost extracted completely, increases extraction times, can only make energy consumption increase, and percentage extraction is constant.Experiment shows that extraction times is 3 times, and extraction efficiency is the highest.

1.4 orthogonal tests: according to experiment of single factor result, the purslane oil extraction yield of take is experimental index, and Three factors-levels test is set, and selects L ₉(3 ⁴) orthogonal table tests.In triplicate, result is got its mean value in each experiment combination.Orthogonal test level of factor table is in Table 1-1, and test-results and analysis of variance table are in Table 1-2 and table 1-3.

Table 1-1 orthogonal test level of factor table

Table 1-2 test quadrature result table

Table 1-3 subcritical abstraction orthogonal test analysis of variance table

Factor	Sum of square of deviations	Degree of freedom	F ratio	F threshold value	Significance
						A	0.184	2	26.286	19.000	*
B	0.027	2	3.857	19.000
						C	0.475	2	67.857	19.000	*
Error	0.02	2	1.000

Note: * is remarkable (P < 0.05)

By table 1-2 and table 1-3, can be found out, extraction temperature is minimum on the extraction yield impact of purslane oil, and extraction time and extracting pressure are remarkable on the impact of purslane oil extraction yield, and extraction temperature is not remarkable on the impact of extraction yield.According to above analysis, can obtain factor affects order and is: extraction time > extracting pressure > extraction temperature.Test-results shows, subcritical fluid extraction purslane oil optimum process condition is A ₂b ₃c ₃, extracting pressure is 0.8MPa, and extraction temperature is 40 ℃, and extraction time is 150min, and amplifying confirmatory experiment purslane oil extraction yield is 2.91%, and result is reproducible, illustrates that selected condition is reasonable.

In 1.5 purslane oil, the composition of lipid acid adopts common GC-MS analytical method.

1.5.1 the physical and chemical index measurement result of grease is in Table 1-4: the physico-chemical property comparison of table 1-4 subcritical fluid extraction and the purslane oil of ultrasonic enzymolysis-Soxhlet extraction

From table, 1-4 learns, the purslane oil quality that subcritical fluid extraction obtains extracts purslane oil that coupling method obtains and the quality of purslane seed oil apparently higher than the ultrasonic enzymolysis of common employing-Soxhlet, the acid value of the purslane oil that ultrasonic enzymolysis-Soxhlet extraction coupling legal system is standby is obviously higher, reaches 10.5 mgKOHg ^-1.Acid value is high may be relevant with oil extracting methods, extract temperature 70 ℃ of left and right, make its destroyed degree of fatty acid oil glyceride structure high, thereby the free fatty acids producing in putting forward oily process is many, causes ultrasonic enzymolysis-Soxhlet to extract the standby purslane oleic acid value of coupling legal system very high.

1.5.2 the lipid acid component analysis of grease is in Table 1-5: the fatty acid compositional analysis of table 1-5 subcritical fluid extraction and the purslane oil of ultrasonic enzymolysis-Soxhlet extraction

From table 1-5, can find out, it is respectively oleic acid, linolic acid, linolenic acid that the purslane oil of ultrasonic assistance enzymolysis-Soxhlet coupling method gained contains 3 kinds of unsaturated fatty acidss, the content of the polyunsaturated fatty acid in hence one can see that purslane oil accounts for 56.39% of fatty acid total amount, and main component is linolic acid and two kinds of compositions of linolenic acid.And the linoleic acid plus linolenic acid that the purslane oil of subcritical fluid extraction gained contains accounts for 64.23% of fatty acid total amount, than front method, improved 7.84%, wherein, linolenic acid content is apparently higher than ultrasonic assistance enzymolysis-Soxhlet coupling method of common employing.

conclusion:subcritical Trimethylmethane extraction purslane oil optimum process condition extracting pressure is 0.8MPa, and extraction temperature is 40 ℃, and extraction time is 150min, and purslane oil extraction yield is up to 2.91% with this understanding; The soxhlet extraction adopting with tradition is compared and has been improved 1.2%; Mensuration and fatty acid oil GC-MS by purslane oil physical and chemical index analyze, result shows: adopt the content of polyunsaturated fatty acid of common ultrasonic assistance enzymolysis-Soxhlet coupling method gained purslane oil to reach 56.39%, the content of polyunsaturated fatty acid of the subcritical Trimethylmethane extraction purslane oil that the present invention adopts reaches 64.23%, than front method, has improved 7.84%.It is low that the purslane oleic acid value of subcritical Trimethylmethane extraction and peroxide value are extracted coupling method than the ultrasonic enzymolysis of common employing-Soxhlet, and quality will be got well.

embodiment five: the preparation of mixed fatty acid before the enrichment of purslane oil

1.1 test methods: accurately take a certain amount of purslane oil being obtained by above-described embodiment four subcritical fluid extractions, add the ethanolic soln of finite concentration KOH, be then put in stirred in water bath, the backflow certain hour of preset temperature, obtain saponification liquor.Add appropriate ice distilled water cooling rapidly, then use 30mL petroleum ether extraction unsaponifiables, extract three times.The water gaging dilution that doubles of water intaking layer, slowly adds 10% HCl, is adjusted to pH=2 ~ 3.Now can obviously see oil reservoir and water layer, use immediately the petroleum ether extraction mixed fatty acid of 20mL, extracting twice, obtains petroleum ether layer by separating funnel.Petroleum ether layer is washed till neutrality by 5% NaCl solution water, then with rotary evaporation after anhydrous sodium sulfate drying, filtration, obtains the front mixed fatty acid of enrichment.Be placed in refrigerator and cooled and freeze storage, stand-by.

Mixed fatty acid yield method of calculation before 1.2 enrichments: mixed fatty acid yield=

In formula: m ₃for the quality (g) of purslane oil, m ₄quality (g) for mixed fatty acid.

1. 3 single factor experiments

1.3.1 KOH-alcohol concn on enrichment before the impact of mixed fatty acid yield

By testing sequence, get respectively KOH-alcohol concn 0.5mol/L, 1mol/L, 1.5mol/L, 2mol/L, 2.5mol/L saponification time 40min, 60 ℃ of saponification temperatures, carry out saponification test to purslane oil.

From accompanying drawing 6, along with the increase gradually of the concentration of KOH-ethanolic soln, before enrichment, the yield of mixed fatty acid presents and first increasing the summary downward trend that tends to be steady afterwards.When KOH-ethanolic soln concentration reaches 1mol/L, before enrichment, the yield of mixed fatty acid is the highest.Because KOH-ethanolic soln can be used as the catalysis of saponification reaction, so the consumption of catalyzer is larger, the speed of saponification reaction is just faster, and saponification is also just more complete.But during the excessive concentration of KOH-ethanolic soln, can cause whole system viscosity to increase, and strengthen the difficulty of saponification reaction, cause reaction not exclusively.Can see, when KOH-alcohol concn surpasses 1mol/L, before enrichment, mixed fatty acid yield declines.

1.3.2 saponification temperature on enrichment before the impact of mixed fatty acid yield

By testing sequence, get respectively 60 ℃ of saponification temperatures, 65 ℃, 70 ℃, 75 ℃, 80 ℃, KOH-alcohol concn 1mol/L, saponification time 40min, carries out saponification test to purslane oil.

From accompanying drawing 7, before enrichment, mixed fatty acid yield raise and increases with temperature before 70 ℃, and the rising with temperature after 70 ℃ tends towards stability.Due to before 70 ℃, with temperature, raise, the corresponding increase of saponification degree, during to 70 ℃, hydrolysis has reached balance, and saponification completes substantially, and before enrichment at this moment, mixing fat acid yield is the highest.So the saponification temperature of purslane oil is selected to 70 ℃ better.

1.3.3 saponification time on enrichment before the impact of mixed fatty acid yield

By testing sequence, get respectively saponification time 20min, 40min, 60min, 80min, 100min, KOH-alcohol concn 1mol/L, 70 ℃ of saponification temperatures, carry out saponification test to purslane oil.

From accompanying drawing 8, along with the prolongation of saponification time, before enrichment, after the increase that must take the lead in of mixed fatty acid, slightly reduce.Saponification is minute three periods generally, and first paragraph time oil mixes with KOH-ethanol, and now two solution are inconsistent, and the second segment time, speed was sharply accelerated when reaction reaches 20%, and the 3rd section of saponification completes.Saponification time is larger, and purslane oil is higher by saponification degree, mixes fat acid just more before the enrichment correspondingly obtaining.After saponification reaction 100min, purslane oil is substantially complete by saponification, then extends the reaction times, and curve is reduction trend.So select 100min better to the saponification time of purslane oil.

1.4 orthogonal tests: according to experiment of single factor result, the mixed fatty acid yield before enrichment of take is experimental index, and Three factors-levels test is set, and selects L ₉(3 ⁴) orthogonal table tests.In triplicate, result is got its mean value in each experiment combination, finally obtains preparing the front mixed fatty acid optimised process of enrichment.

Orthogonal test level of factor table is in Table 2-1, and test-results and analysis of variance table are in Table 2-2 and table 2-3.

Table 2-1 orthogonal test level of factor table

Table 2-2 test quadrature result table

Table 2-3 orthogonal test analysis of variance table

Factor	Sum of square of deviations	Degree of freedom	F ratio	F threshold value	Significance
						A	74.563	2	19.815	19.000	*
B	3.763	2	1.000	19.000
						C	137.637	2	36.576	19.000	*
Error	3.76	2

Note: * is remarkable (P < 0.05)

By table 2-2 and 2-3, can be found out, in extreme difference value, maximum is saponification time, is secondly KOH-alcohol concn, is finally saponification temperature, and it is C > A >B that its factor affects order.Saponification optimal conditions is A ₁b ₃c ₃, KOH-alcohol concn is 0.5 mol/L, and saponification temperature is 75 ℃, and saponification time is 120min, and the front mixed fatty acid yield of enrichment is up to 57.36% with this understanding.

The physical and chemical index of 1.5 greases is measured: peroxide value: measure with reference to GB/T5538-2005 peroxide value of vegetable oil; Acid value: measure with reference to GB/T5530-2005 acid value of plant oil; Iodine value: measure with reference to GB/T5532-2008 vegetables oil iodine value; Saponification value: with reference to GB/T5534-2008 saponification value of lipids assay method; Before enrichment, in mixed fatty acid, lipid acid composition GC-MS analytical procedure adopts common method.

1.5.1 the physical and chemical index measurement result of mixed fatty acid before enrichment

The physical and chemical index of mixed fatty acid before table 2-4 enrichment

Mensuration project	Annex seven mixed fatty acids
		Smell	The distinctive fragrance of purslane oil
Color and luster	Deep yellow
		Peroxide value/meqkg ^-1	0.92
Acid value/mgKOHg ^-1	2.78
		Saponification value/mgKOHg ^-1	201.03
Iodine number/g100g ^-1	86.71

From table, 2-4 learns, before enrichment, color and luster and the purslane oil phase of mixed fatty acid are superficial, and approaches at normal temperatures liquid.

1.5.2 before enrichment, the lipid acid main component of mixed fatty acid is analyzed

Before table 2-5 enrichment, the lipid acid main component of mixed fatty acid is analyzed

Peak number	Relative molecular mass	Corresponding lipid acid	Molecular formula	Relative content (%)
					1	242	Myristic acid	C14:0	0.21
2	270	Palmitinic acid	C16:0	17.01
					3	298	Stearic acid	C18:0	4.31
4	296	Oleic acid	C18:1	10.01
					5	294	Linolic acid	C18:2	35.31
6	292	Linolenic acid	C18:3	32.81
					7	326	Eicosanoic acid	C20:0	0.51
8	354	Behenic acid	C21:0	—
					9	282	Phthalic acid dimethoxy	C ₁₄H ₁₈O ₆	1.46

By table, 2-5 can find out, before the enrichment preparing, in mixed fatty acid, detect 8 kinds of lipid acid in purslane oil, saturated fatty acid comprises myristic acid, palmitinic acid, stearic acid, eicosanoic acid, behenic acid, monounsaturated fatty acids comprises oleic acid, polyunsaturated fatty acid accounts for 68.12%, is mainly linolic acid, linolenic acid.

conclusion:the present embodiment is by adopting saponification to prepare mixed fatty acid before enrichment to purslane oil, the mixed fatty acid yield before enrichment of take has been investigated KOH-alcohol concn, saponification temperature, the impact of saponification time three factors on saponification reaction as index, by design orthogonal test, obtains optimal processing parameter; The optimum process condition of saponification is: KOH-alcohol concn is 0.5 mol/L, and saponification temperature is 75 ℃, and saponification time is 120min, and the front mixed fatty acid yield of enrichment is up to 57.36% with this understanding.Polyunsaturated fatty acid before enrichment in mixed fatty acid accounts for 68.12%, is mainly linolic acid, linolenic acid.

embodiment six: urea clathration enriching polyunsaturated fatty acid technique

The present embodiment adopts urea adduct method enriching polyunsaturated fatty acid, pass through decrease temperature crystalline, saturated fatty acid in mixed fatty acid before enrichment and oleic acid are removed to the polyunsaturated fatty acid of purifying wherein, thereby mixed fatty acid after the enrichment that enrichment obtains containing highly purified polyunsaturated fatty acid.

1.1 test materialss: before enrichment, mixed fatty acid: embodiment five preparations provide.

1.2 urea adduct method enriching polyunsaturated fatty acid technical process: get a certain amount of urea and dehydrated alcohol and be placed in round-bottomed flask, heated and stirred refluxes at 60 ℃, until urea all dissolves.Pour into rapidly in the three-necked flask that fills the front mixed fatty acid of a certain amount of enrichment, after mixing, heated and stirred backflow certain hour at 60 ℃, be cooled to room temperature, be placed in after 4 ℃ of refrigerator crystallization certain hours, by the rapid suction filtration of crystal clathrate, obtain crystal clathrate and filtrate two portions, wherein in crystal clathrate, contain most saturated fatty acids and oleic acid, the polyunsaturated fatty acid containing in filtrate thereby obtain enrichment.

The not extraction of inclusion lipid acid (containing high purity polyunsaturated fatty acid) in filtrate: obtain solid particulate after filtrate rotary evaporation is removed to ethanol, add 5mL water, dense HCL is adjusted to pH5 ~ 6, adding appropriate sherwood oil extracts again, till extremely using urea detection paper without urea with distillation washing ether layer, use anhydrous sodium sulfate drying 4h, mixed fatty acid after the enrichment that can enrichment obtains containing highly purified polyunsaturated fatty acid.

The iodine value of mixed fatty acid and the mensuration of the rate of recovery after 1.3 enrichments

After enrichment, in mixed fatty acid, iodine value adopts GB/T5532-2008 vegetables oil iodine value assay method to measure, its rate of recovery calculation formula:

Mixed fatty acid quality * 100% before mixed fatty acid quality/enrichment after the rate of recovery=enrichment

The forward and backward mixed fatty acid mass unit of enrichment in formula is g.

1.4 single factor experiment

1.4.1 before urea/enrichment, mixed fatty acid (m/m) affects mixed fatty acid urea clathration after enrichment

Get respectively the front mixed fatty acid (m/m) of urea/enrichment: 0.5,1.0,1.5,2.0,2.5, fixedly before enrichment, mixed fatty acid/ethanol (m/v) is 1:18, return time 20min, the inclusion time is 24h, with the rate of recovery and the iodine number of mixed fatty acid after enrichment, is used as criterion.

By accompanying drawing 9, can be found out, along with the increase of urea content, iodine number presents first increases the trend reducing afterwards, and the rate of recovery presents the rule that falls progressively.When the consumption of urea is very few, urea clathration saturated fatty acid is incomplete, thereby has more saturated fatty acid in the lipid acid obtaining, and causes iodine number lower; Continue to strengthen amount of urea, with saturated fatty acid, constantly by urea clathration, so the rising of iodine value has also illustrated that after enrichment, the purity of the polyunsaturated fatty acid in mixing-in fat raises.When mixed fatty acid before urea/enrichment (m/m) reaches 1, iodine number reaches maximum value.And when continuing to strengthen amount of urea, it is except energy inclusion saturated fatty acid and monounsaturated fatty acids (oleic acid), the composition that also has the polyunsaturated fatty acids such as linoleic acid plus linolenic acid of part is formed crystallization by urea clathration, can cause the rate of recovery of the polyunsaturated fatty acid in mixed fatty acid after enrichment greatly to reduce, corresponding purity also can reduce; And addition is too much, form the excessive difficulty that too much can increase filtration of crystal, these factors all can affect the purity of product.Consider, it is 1 suitable selecting before urea/enrichment mixed fatty acid (m/m).

1.4.2 before enrichment, mixed fatty acid/ethanol (m/v) affects mixed fatty acid urea clathration after enrichment

Get respectively the front mixed fatty acid/ethanol (m/v) of enrichment: 1:12,1:14,1:16,1:18,1:20, fixedly before urea/enrichment, mixed fatty acid (m/m) is 1.0, return time 20min, the inclusion time is 24h, with the rate of recovery and the iodine number of mixed fatty acid after enrichment, is used as criterion.

In urea clathration process, after enrichment, the purity of mixed fatty acid and the rate of recovery can directly be subject to the impact of urea addition, as the addition of the ethanol of solvent, have been directly connected to urea and by the degree of scatter between the saturated and monounsaturated fatty acids of inclusion.By accompanying drawing 10, can be found out, iodine number along with enrichment before mixed fatty acid/ethanol (m/v) increase and reduce purity drop.And the rate of recovery in contrast, in rising trend.Reaction has just started urea and can not dissolve completely and cause inclusion degree bad, but mixed fatty acid/ethanol (m/v) raises gradually before enrichment, amount of urea is also just with respect to increase, and polyunsaturated fatty acid that also can inclusion part, causes the effect of inclusion undesirable.When mixed fatty acid/ethanol before enrichment (m/v) reaches 1:16, after enrichment, the purity of mixed fatty acid and the rate of recovery have reached optimum value separately jointly.

1.4.3 the selection of return time on enrichment after mixed fatty acid urea clathration impact: get respectively return time: 10min, 20min, 30min, 40min, 50min, fixedly before urea/enrichment, mixed fatty acid (m/m) is 1.0, before enrichment, mixed fatty acid/ethanol (m/v) is 1:16, the inclusion time is 24h, with the rate of recovery and the iodine number of mixed fatty acid after enrichment, is used as criterion.

The main purpose refluxing is that the same urea-ethanolic soln of mixed fatty acid before enrichment is mixed, and makes carrying out smoothly of urea clathration reaction.If the return stirring time is short, to mix inhomogeneously, the quality product obtaining can decrease.Experiment shows, return time on enrichment after purity and the rate of recovery impact of mixed fatty acid little.But can find out, return time is that the rate of recovery that obtains of 10min is later lower than backflow 20min.And return time surpasses after 20min, little on urea clathration reaction impact.But return time is long, cause power consumption large, also increased on the other hand polyunsaturated fatty acid and caused rotten risk, so select return time 20min to be advisable.

1.4.4 the selection of inclusion time on enrichment after mixed fatty acid urea clathration impact

Get respectively inclusion time: 6h, 12h, 18h, 24h, 30h, fixedly before urea/enrichment, mixed fatty acid (m/m) is 1.0, mixed fatty acid/ethanol (m/v) is 1:16 before enrichment, and return time 20min is used as criterion with the rate of recovery and the iodine number of mixed fatty acid after enrichment.

By accompanying drawing 11, can obviously be found out, iodine value is along with the inclusion time increases and present the trend that first raises and reduce afterwards gradually, reason may be because urea clathration reaction crystal grain of the inclusion compound of urea when just starting is less, thorough separating saturated fatty acid during suction filtration, make iodine value lower, and along with the inclusion time increases gradually, it is large that the crystal grain of inclusion compound become gradually, the effect of inclusion saturated fatty acid is become better and better, and iodine value is increased gradually.But when inclusion overlong time, after saturated fatty acid and oleic acid are all completed by urea clathration, understand some polyunsaturated fatty acid and enter inclusion phase, after enrichment, the rate of recovery of mixed fatty acid reduces the reduction of the purity that causes product.So select inclusion time iodine value when 24h to reach maximum value.

1.5 orthogonal tests: according to experiment of single factor result, adopt L ₉(3 ⁴) orthogonal table, do the front mixed fatty acid (m/m) of urea/enrichment, the front mixed fatty acid/ethanol (m/v) of enrichment, return time and inclusion times four factor three horizontal quadratures experiments, each is tested in triplicate, and result is got its mean value.With orthogonal design assistant II v3.1.1 statistical software, carry out interpretation of result.

Orthogonal test level of factor table is in Table 3-1, and test-results and analysis of variance table are in Table 3-2 and table 3-3,3-4.

Table 3-1 orthogonal test level of factor table

Table 3-2 urea clathration orthogonal experiments table

Table 3-3 urea clathration rate of recovery analysis of variance table

Factor	Sum of square of deviations	Degree of freedom	F ratio	F threshold value	Significance
						A	18.078	2	18.024	19.000
B	21.504	2	21.440	19.000	*
						C	47.426	2	47.284	19.000	*
Error	1.00	2

Note: * is remarkable (P < 0.05)

Table 3-4 urea clathration iodine number analysis of variance table

Factor	Sum of square of deviations	Degree of freedom	F ratio	F threshold value	Significance
						A	73.510	2	45.715	19.000	*
B	24.971	2	15.529	19.000
						C	69.335	2	43.119	19.000	*
Error	1.61	2

Note: * is remarkable (P < 0.05)

By table 3-2 and 3-3, can be found out, what the rate of recovery of mixed fatty acid after enrichment was had the greatest impact is the inclusion time, next is mixed fatty acid/ethanol (m/v) before enrichment, be finally mixed fatty acid (m/m) before urea/enrichment, it is the front mixed fatty acid of mixed fatty acid/ethanol (m/v) urea/enrichment before the > enrichment of inclusion time that its factor affects order.Its optimal conditions is A ₂b ₂c ₂, i.e. before urea/enrichment, mixed fatty acid (m/m) is 1, and before enrichment, mixed fatty acid/ethanol (m/v) is 1:16, and the inclusion time is 24h.By table 3-2 and table 3-4, can be found out, it is mixed fatty acid (m/m) before urea/enrichment that iodine number is had the greatest impact, next is the inclusion time, be finally mixed fatty acid/ethanol (m/v) before enrichment, it is before mixed fatty acid (m/m) > inclusion time > enrichment, to close lipid acid/ethanol (m/v) before urea/enrichment that its factor affects order.Iodine number optimal conditions is A ₂b ₂c ₂, i.e. before urea/enrichment, mixed fatty acid (m/m) is 1, and before enrichment, mixed fatty acid/ethanol (m/v) is 1:16, and the inclusion time is 24h.Consider primary and secondary influence factor, the final optimum combination of selecting is: A ₂b ₂c ₂, i.e. before urea/enrichment, mixed fatty acid (m/m) is 1, and before enrichment, mixed fatty acid/ethanol (m/v) is 1:16, and the inclusion time is 24h.With this understanding, the final product rate of recovery is up to 78.04%, and iodine number is up to 112.03 g/100g.

The physical and chemical index of 1.6 greases is measured: adopt common measuring method.

1.6.1 the physical and chemical index comparison of mixed fatty acid after mixed fatty acid and enrichment before purslane oil, enrichment

The physical and chemical index comparison of mixed fatty acid after mixed fatty acid and enrichment before table 3-5 subcritical abstraction purslane oil, enrichment

Mensuration project	Purslane oil	Mixed fatty acid before enrichment	Mixed fatty acid after enrichment
				Smell	The distinctive fragrance of purslane oil	The distinctive fragrance of purslane oil	The distinctive fragrance of purslane oil
Color and luster	Yellow-green colour	Deep yellow	Light yellow
				Peroxide value/meqkg ^-1	2.3	0.92	0.19
Acid value/mgKOHg ^-1	4.18	2.78	1.15
				Saponification value/mgKOHg ^-1	164.34	201.03	150.32
Iodine number/g100g ^-1	79.07	86.71	112.03

From table, 3-5 learns, after the enrichment being obtained by urea adduct method enrichment, mixed fatty acid is compared first two product, its peroxide value, acid number, saponification value are starkly lower than mixed fatty acid before purslane oil and enrichment, and iodine number is apparently higher than mixed fatty acid before purslane oil and enrichment.The present invention utilizes in the process of mixed fatty acid after the enrichment that urea adduct method enrichment obtains and removes impurity, and the oleic acid composition that after the enrichment obtaining, mixed fatty acid contains minute quantity, so quality is better than the quality of purslane oil and the front mixed fatty acid of enrichment.

1.6.2 mixed fatty acid comparison of ingredients after mixed fatty acid and enrichment before purslane oil, enrichment

The Analysis of Fatty Acids Composition comparison of mixed fatty acid before mixed fatty acid and purslane oil, enrichment after table 3-6 enrichment

By table, 3-6 can find out, before purslane oil and enrichment thereof, after mixed fatty acid, enrichment, in mixed fatty acid, mainly detect 8 kinds of lipid acid, in purslane oil, fatty acid carbon chain atomicity is mainly between 14 ~ 21, saturated fatty acid is mainly myristic acid, palmitinic acid, stearic acid, eicosanoic acid etc., monounsaturated fatty acids mainly comprises oleic acid etc., and polyunsaturated fatty acid mainly comprises linolic acid and linolenic acid.After the enrichment being obtained by urea adduct method enrichment, in mixed fatty acid, lipid acid mainly be take linolic acid, linolenic acid as main, and accounts for 94.56% of total amount.After the enrichment obtaining under urea adduct method optimum process condition, mixed fatty acid is compared with the lipid acid of purslane oil, and content of polyunsaturated fatty acid brings up to 94.56% by 64.23%.This illustrates that most saturated fatty acid and monounsaturated fatty acids are formed crystallization by urea clathration, by extraction, remove by filter crystallisate, thereby mixed fatty acid after the enrichment that obtains containing highly purified polyunsaturated fatty acid, not only proving again adopt the superiority of urea adduct method enriching polyunsaturated fatty acid, also for theoretical basis has been established in the exploitation in polyunsaturated fatty acid market.

conclusion:the present invention takes into full account in design technology project, and urea forms 0.4 ~ 0.5nm hexagon tubulose crystallizer road internal diameter by intermolecular hydrogen bonding, and its internal diameter has limited the molecule cross-sectional sizes of its inclusion compound, and linear saturated fatty acids molecule cross section is little easily by inclusion; And that unsaturated fatty acids is affected by two keys is larger, molecule cross section is greater than urea crystals internal diameter of the pipeline, and difficult quilt contains wherein, and two key is more much more difficult.So saturated fatty acid and the oleic acid of urea in can inclusion lipid acid.When temperature reduces, urea and saturated and monounsaturated fatty acids (oleic acid) form crystal inclusion compound and separate out, and adopt the method for filtering to remove the polyunsaturated fatty acid that crystal inclusion compound can obtain higher degree.

It is to judge index that iodine number and the rate of recovery of mixed fatty acid after enrichment take in the present invention, the front mixed fatty acid (m/m) of urea/enrichment, the front mixed fatty acid/ethanol (m/v) of enrichment, return time, the impact of inclusion time on urea clathration reaction have been investigated, by single factor and Orthogonal experiment results, consider primary and secondary influence factor, determine urea clathration optimal conditions, utilize GC-MS to analyze the product fatty acid component after purifying.Result shows: urea clathration optimised process is: take urea/mixed fatty acid (m/m) as 1, mixed fatty acid/ethanol (m/v) is dissolved in ethanolic soln by urea as 1:16, add the front mixed fatty acid backflow 20min of enrichment, be placed on standing 24h at 4 ℃.After the enrichment obtaining under this technique, mixed fatty acid is compared with the lipid acid of purslane oil before inclusion not, and polyunsaturated fatty acid brings up to 94.56% by 64.23% content.

embodiment seven: the preparation of purslane polyunsaturated fatty acid microcapsule

1.1 polyunsaturated fatty acid micro capsule technology flow processs: by the appropriate wall material taking, according to gum arabic: beta-cyclodextrin: modified starch=1:1:2 stirs 30min under 60 ℃ of conditions, then be placed on magnetic stirring apparatus and stir, it is fully dissolved, and placement room temperature is cooling, obtains water.Press wall material: core=3:1 proportioning, take mixed fatty acid after enrichment prepared by above-described embodiment, and add a certain amount of tween 80 to carry out stirring and dissolving under 60 ℃ of conditions, obtain oil phase.Oil phase is slowly joined in water and continues to stir under 60 ℃ of conditions, be placed on magnetic stirring apparatus and stir, it is fully dissolved, then with high speed homogenization refiner, under 10000r/min condition, disperse 3min, make oil droplet uniformly emulsify in wall material solution, be milk sap state, be finally placed in the dry 12h of vacuum freezing drying oven of-55 ℃, collect product.

1.2 microcapsule show the mensuration of oil length and the calculating of embedding rate: use sherwood oil as solvent, accurately take 5g product, 50ml sherwood oil is divided 3 times to add, all vibrations, filter merging filtrate at every turn, filtrate rotary evaporation is removed to moisture, dry to constant weight, be the quality of product surface oil, be i.e. surface of microcapsule oil-contg.

Surface oil length=extract fuel-displaced quality (g)/sample quality (g) * 100%

Embedding rate=(1-surface oil length) * 100%

With analytical balance, accurately take 5.000g microcapsule product, the sherwood oil of take extracts its surface oil as solvent.According to formula as calculated, purslane polyunsaturated fatty acid microcapsule product surface oil length prepared by the present invention is 17.3%, and embedding rate is 82.7%;

1.3 Microcapsules Size analyses: purslane polyunsaturated fatty acid microscapsule powder prepared by a little the present invention is spread in the sample table of having pasted double faced adhesive tape, blows away unnecessary powder; After metal spraying, with the surface tissue of sem observation microcapsule product, acceleration voltage is 20KV.

Microcapsules Size is analyzed: the wall material of microcapsule is embedding core effectively, is one of important indicator of microcapsule product success or not, can be by the observation of surface of microcapsule structure is judged.Microcapsule are placed in to its size distribution of micro-Microscopic observation, median size is approximately distributed between 4～10 μ m, median size is 7.5 μ m, reason may be that high speed homogenization refiner that this experiment adopts carries out emulsification and makes purslane polyunsaturated fatty acid and wall material thereof in solution, be dispersed into drop more tiny, homogeneous, and then impel more polyunsaturated fatty acid core to be wrapped in wall material, improved embedding rate.

1.4 microcapsule product sensory evaluations: microcapsule product is placed under available light, take blank sheet of paper as background, detect by an unaided eye its color and luster, form, smell its smell.

Sensory evaluation: the microcapsule product color of purslane polyunsaturated fatty acid is milk yellow, without special odor, product drying, powder is evenly fine and smooth, and mobility is better.

The method of preparing purslane polyunsaturated fatty acid microcapsule from purslane slag provided by the invention; the advantage of this method is that required equipment is simple, easy and simple to handle; production cost is lower; especially under hot environment, do not carry out, can almost entirely protect the molecular structure of unsaturated fatty acids and physico-chemical property unaffected.

Prepare purslane polyunsaturated fatty acid microcapsule and it is carried out to quality evalution through above-described embodiment, result shows, microcapsule product color is milk yellow, and without special odor, powder is evenly fine and smooth; Surface oil length is 17.3%, and embedding rate is 82.7%; Microcapsule water ratio is 3.80%, meets the requirement of microcapsule moisture content; Median size is 306.6nm.

embodiment eight: the preparation of purslane polyunsaturated fatty acid soft capsule

1.1 polyunsaturated fatty acid micro capsule technology flow processs: by the appropriate wall material taking, according to gum arabic: beta-cyclodextrin: modified starch=1:1:2 stirs 30min under 60 ℃ of conditions, then be placed on magnetic stirring apparatus and stir, it is fully dissolved, and placement room temperature is cooling, obtains water.Press wall material: core=3:1 proportioning, take mixed fatty acid after enrichment prepared by above-described embodiment, and add a certain amount of tween 80 to carry out stirring and dissolving under 60 ℃ of conditions, obtain oil phase.Oil phase is slowly joined in water and continues to stir under 60 ℃ of conditions, be placed on magnetic stirring apparatus and stir, it is fully dissolved, then with high speed homogenization refiner, under 10000r/min condition, disperse 3min, make oil droplet uniformly emulsify in wall material solution, be milk sap state, standby; In glue pot, first add glycerine (15%), pure water (40%), add gelatin (45%) after stirring and be heated to 75 ℃, stirring and dissolving 60min dissolves gelatin completely, vacuumizes, and removes bubble, 60 ℃ of heat preservation for standby use; Glue is packed in capsule machine processed and carries out pelleting with the content mixing, seasoning after capsule and pill sizing, temperature is controlled at 45 ℃～50 ℃, prepares purslane polyunsaturated fatty acid soft capsule.

1.2 content uniformities: with reference to < < Pharmacopoeia of People's Republic of China > > 2010 editions test procedure disintegration, the content uniformity of sample is all in the limits of regulation.

1.3 external disintegrations are investigated: with reference to < < Pharmacopoeia of People's Republic of China > > 2010 editions test procedure disintegration, the disintegration of sample all within the limits prescribed.

1.4 soft capsule product sensories are evaluated: soft capsule product is placed under available light, take blank sheet of paper as background, detect by an unaided eye its color and luster, form, smell its smell.

the method of preparing purslane polyunsaturated fatty acid soft capsule from purslane slag provided by the invention; its advantage is that required equipment is simple, easy and simple to handle; production cost is lower; especially under hot environment, do not carry out, can almost entirely protect the molecular structure of unsaturated fatty acids and physico-chemical property unaffected.

Through above-described embodiment, prepare purslane polyunsaturated fatty acid soft capsule and it is carried out to quality evalution and show, soft capsule product colour is faint yellow, and without special odor, granularity is moderate, and content uniformity and disintegration are all in specialized range.

embodiment nine: the effect of lowering blood sugar experiment of purslane polyunsaturated fatty acid microcapsule and soft capsule

Purslane is as a kind of conventional vegetables no side effects, and all there is supply in the four seasons, originates very abundant.The dosage of this experiment setting purslane polyunsaturated fatty acid microcapsule and soft capsule is consistent, the impact of research various dose on the function of blood sugar reduction of diabetes rat.Rat is divided into 6 groups at random, is respectively Normal group, positive controls, model control group, low dose group, middle dosage group, high dose group, measure body weight and blood sugar and insulin level each side index.By the experiment of rat function of blood sugar reduction, confirm: purslane polyunsaturated fatty acid microcapsule and soft capsule have the effect of hypoglycemic blood.Hence one can see that, and purslane polyunsaturated fatty acid microcapsule and soft capsule can effectively reduce blood sugar and the insulin level of diabetes rat, prevention and treatment for diabetes have positive effect, get a good chance of becoming novel healthy food or even the medicine that regulates blood sugar.

1.1 test reagents: purslane polyunsaturated fatty acid microcapsule and soft capsule prepared by above-described embodiment.

1.2 test apparatuses: the happy Kang Quan blood glucose meter of Roche and blood sugar test paper (Irish Roche Holding Ag); PW-3 digital display balance (Kaishi Electronic Co., Ltd., Shanghai); The common device facilities such as irrigation stomach device (Shanghai Medicine Laser Instrument Plant).

1.3 animal experiment method

1.3.1 animal: 60 of clean level SD male rats, body weight 180 ~ 220g, is provided by Xinjiang Medicine University's experimentation on animals center, SCXK(is new for laboratory animal credit number) 2011-0003.

1.3.2 feed: basal feed is that market is common.

1.3.3 animal divides into groups and experimental technique: 60 SD male rats adapt to and feed after one week at basal feed, by body weight, choose at random 10 as Normal group, and all the other 50 is modeling group.Streptozotocin is dissolved in the citric acid-sodium citrate damping fluid of the 0.1mol/L pH4.4 of existing preparation before use, is mixed with 1% concentration, is formulated in ice bath and carries out.Before injection, water 18h is can't help in all rat fasting.Modeling group is pressed 60mg/kg dosage abdominal injection 1%STZ solution, the citric acid-sodium citrate damping fluid of Normal group abdominal injection same volume.After 3d, water 10h is can't help in fasting, through tail vein blood, surveys fasting plasma glucose, take fasting plasma glucose >=16.7mmol/L as modeling success.Method: first use cotton ball soaked in alcohol wiping afterbody, make tail venous engorgement, cut osculum with knife blade, outflow one is bled and dropped on blood sugar test paper, inserts blood glucose meter, records blood glucose value after 5s.While repeatedly measuring, the edge of a knife is moved to near-end by far-end.Take fasting plasma glucose >=16.7mmol/L as standard, be defined as diabetes model.Before surveying blood sugar, be subject to the equal fasting 10h of laboratory animal.

Modeling success rat is divided into 5 groups of model control group, low dose group, middle dosage group, high dose group, positive controls, 10 every group at random by body weight and blood glucose value.Low dose group, middle dosage group, high dose group are respectively: 32mg/(kgbw ^-1), 64mg/(kgbw ^-1), 128mg/(kgbw ^-1), the solution with distilled water preparation of difference gavage various dose, Normal group and model control group gavage distilled water, gavage volume is 10ml/(kgbw ^-1), 1/d.Positive controls gavage is with the glyburide 80mg/(kgbw of distilled water preparation ^-1), 1/d.

Gavage is 4 weeks continuously, respectively at measuring after administration 14,28d, detects index.

Detect index: body weight, fasting plasma glucose, insulin level.

Fasting plasma glucose: after last administration, water 10h is can't help in fasting, surveys fasting plasma glucose through tail vein blood, first use cotton ball soaked in alcohol wiping afterbody, make tail venous engorgement, with knife blade, cut osculum, outflow one is bled and is dropped on blood sugar test paper, inserts blood glucose meter, records blood glucose value after 5s.While repeatedly measuring, the edge of a knife is moved to near-end by far-end.

Insulin level: after last administration, water 10h is can't help in fasting, rat is through etherization, by abdominal aortic blood, separation of serum, reference reagent box specification sheets is with serum measured by radioimmunoassay insulin content.

1.3.4 statistical procedures: adopt SPSS 11.5 softwares to carry out statistical procedures, data represent with x ± s, relatively adopt one-way analysis of variance between group. p<0.05 has statistical significance, significant difference.

1.4 result

1.4.1 purslane polyunsaturated fatty acid microcapsule and the soft capsule impact on SZT diabetes rat body weight, respectively at before modeling, after modeling, after administration two weeks and survey the body weight of rat after administration surrounding, obtains result as following table 4-1:

Note: " * " represents to compare with model group p<0.05 level difference is remarkable; " * * " represents to compare with model group p<0.01 level difference is remarkable.

From table 4-1, Normal group and model control group can find out that the marked difference of diabetes rat and normal rat is weight loss, and relatively whether the variation of perfusion front and back body weight can have effect of lowering blood sugar by part illustrative experiment medicine.Experimental result shows: after modeling, rat becomes thin gradually, weight loss.Significant difference between high dose group and diabetic model group group ( p<0.05), prove that purslane polyunsaturated fatty acid microcapsule and soft capsule have remarkable restitution to diabetes model rat body weight.Low dose group also has restitution to diabetes rat body weight, but DeGrain.

1.4.2 purslane polyunsaturated fatty acid microcapsule and soft capsule on the impact of SZT blood glucose in diabetic rats and insulin level in Table 4-2.

From table 4-2, purslane polyunsaturated fatty acid microcapsule and soft capsule have certain hypoglycemic activity to SZT diabetes rat.3d after modeling, each is organized blood glucose value and compares with Normal group, have utmost point significant difference ( p<0.01), prove the hyperglycemia of successfully having induced rat with streptozotocin.After administration, model control group blood glucose value does not only reduce, and even also slightly raises; And positive controls and high dose group to SZT diabetes rat produced significant hypoglycemic activity ( p<0.05), the average blood sugar value after high dose group administration has reduced by 51.04% before than administration, compares and has reduced by 37.93% with model control group, there were significant differences ( p<0.05); And low dose group also has certain hypoglycemic activity, but contrast with model compare there was no significant difference ( p>0.05).Meanwhile, purslane polyunsaturated fatty acid microcapsule and soft capsule have dose-dependently aspect hypoglycemic, and high dose group has significant blood sugar reducing function.

SZT rat model serum insulin content is starkly lower than normal rat, high dose group than model group significantly raise serum insulin content ( p<0.01), and middle dosage group also relatively improved serum insulin content ( p<0.015), result shows, purslane polyunsaturated fatty acid microcapsule and soft capsule, except hypoglycemic activity, still have the effect that stimulates B cell to discharge serum insulin.

Conclusion: this experiment shows that purslane polyunsaturated fatty acid microcapsule and soft capsule are falling the hypoglycemic while, the content of the diabetes rat serum insulin that also can raise, with model group rat relatively have significant difference ( p<0.01).Visible, the present invention is tested and is confirmed that purslane polyunsaturated fatty acid microcapsule and the soft capsule of preparation can effectively reduce blood sugar and insulin level by diabetes rat, prevention and treatment for diabetes have positive effect, show that purslane polyunsaturated fatty acid microcapsule provided by the invention and purslane polyunsaturated fatty acid soft capsule are with a wide range of applications and prospect in the food of preparation prevention and treatment diabetes or medicine.

Above-described embodiment is only for example of the present invention is clearly described, and the not restriction to embodiment.For those of ordinary skill in the field, can also make other changes in different forms on the basis of the above description.Here exhaustive without also giving all embodiments.And the apparent variation of being amplified out thus or change are still among protection scope of the present invention.

Claims

1. from purslane slag, prepare a method for purslane polyunsaturated fatty acid microcapsule, it is characterized in that, % is according to weight percent meter, and the concrete steps of purslane polyunsaturated fatty acid are as follows:

(1) preparation of purslane slag: take fresh purslane, Drinking Water is main raw material, fresh purslane is selected, clean, chopping, by weight 1:3 ratio, soaks 12h, and infusion 6h at 80 ℃, filters and obtains purslane slag;

(2) adopt the pre-treatment that purslane slag is raw material: the purslane slag that the purslane slag in the processing of employing purslane or above-mentioned steps provide is raw material, through 70 ℃ of dry, pulverize, sieve 60 orders, weighing, fillers;

(3) subcritical Trimethylmethane abstraction technique extracts purslane oil: carry out subcritical fluid extraction technique, impose a condition, extracting pressure 0.8MPa, 40 ℃ of extraction temperature, extraction 150min, by the purslane oil extractum through extraction preparation, wait extract the purslane oil extractum that after three times, merging obtains at every turn, after the centrifugal 15min of 4000r/min, filtration, prepare finished product purslane oil;

(4) saponification is prepared the front mixed fatty acid of enrichment: according to g/mL, count, purslane oil is mixed by 1:20 with 0.5mol/L KOH-ethanolic soln, the purslane oil that soon prepared by above-mentioned steps (3) adds the ethanolic soln of KOH, then be put in stirred in water bath, the backflow 120min of 75 ℃, obtain saponification liquor; Add with the ice distilled water of 0.5 mol/L KOH-ethanolic soln equivalent cooling rapidly, then use the petroleum ether extraction unsaponifiables with 0.5 mol/L KOH-ethanolic soln equivalent, through extraction repeatedly, the water gaging dilution that doubles of water intaking layer, slowly adds 10% HCl, is adjusted to pH=2 ~ 3, now can obviously see oil reservoir and water layer, use immediately the petroleum ether extraction mixed fatty acid with 0.5mol/LKOH-ethanolic soln equivalent, extraction repeatedly, obtains petroleum ether layer by separating funnel; Petroleum ether layer is washed till neutrality by 5% NaCl solution water, then with rotary evaporation after anhydrous sodium sulfate drying, filtration, obtains the front mixed fatty acid of enrichment ;

2. from purslane slag, prepare a method for purslane polyunsaturated fatty acid soft capsule, it is characterized in that, % is according to weight percent meter, and concrete grammar step is as follows:

(2) adopt the pre-treatment that purslane slag is raw material: purslane slag prepared by the purslane slag in the processing of employing purslane or above-mentioned steps is raw material, through 70 ℃ of dry, pulverize, sieve 60 orders, weighing, fillers;

3. from purslane slag, prepare purslane polyunsaturated fatty acid microcapsule prepared by the method for purslane polyunsaturated fatty acid in preparation prevention and the food for the treatment of diabetes or the application medicine as claimed in claim 1.

4. from purslane slag, prepare purslane polyunsaturated fatty acid soft capsule prepared by the method for purslane polyunsaturated fatty acid in preparation prevention and the food for the treatment of diabetes or the application medicine as claimed in claim 2.