CN103479875A - Preparation method and application of high-activity radix polygonati officinalis concentrated solution - Google Patents
Preparation method and application of high-activity radix polygonati officinalis concentrated solution Download PDFInfo
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Abstract
The invention provides a preparation method of a high-activity radix polygonati officinalis concentrated solution, which comprises the following steps: carrying out enzyme hydrolysis reaction on radix polygonati officinalis at 25-45 DEG C for 1-4 hours, wherein the weight ratio of the radix polygonati officinalis to the pure water is 1:(6-12); carrying out dynamic circulating extraction at 85-95 DEG C twice; filtering through a microporous film, wherein the delivery pressure is 0.3-0.6 Mpa, and the pore size of the film is 0.5-1.5 mu m; carrying out primary reduced pressure concentration on the filtrate at 60-75 DEG C under the vacuum degree of -0.06 to -0.08 Mpa until the solid content is 15-25% Brix; sending the concentrated solution into a centrifugal machine to carry out purification and separation by high-speed centrifugation, wherein the rotation speed is 12000-20000 rpm, and the separating factor is 11000-15000; and carrying out secondary reduced pressure concentration at 60-75 DEG C under the vacuum degree of -0.06 to -0.08 Mpa until the solid content in the concentrated solution reaches 60-65%Brix.
Description
Technical field
The invention belongs to and belong to medicine, field of health care food, a kind of preparation method and application thereof of high activity Rhizoma Polygonati Odorati concentrated solution are provided especially.
Background technology
The dry root that Rhizoma Polygonati Odorati is Liliaceae Polygonatum Rhizoma Polygonati Odorati Polygonatumodoratum (Mill.) Druce herbaceos perennial.Be listed in the top grade beginning as Chinese medicine and be stated from Shennong's Herbal, mainly contain the compositions such as fragrant solomonseal rhizome polyoses, steroidal saponin, flavone, tannin, sterol, phlegmatic temperament, alkaloid and cardiac glycoside, the main active that wherein fragrant solomonseal rhizome polyoses, steroidal saponin are Rhizoma Polygonati Odorati, there is nourishing YIN to relieve dryness, the effect of the cough-relieving of promoting the production of body fluid, multiplex in treatment dry throat and mouth, lung stomach-Yin wound, interior-heat quench one's thirst, the diseases such as vexed cardiopalmus, cough caused by dryness-heat, pachylosis, diabetes are had to good therapeutic effect.What Rhizoma Polygonati Odorati was put into the Ministry of Public Health promulgation is in the article list of food and medicine, safe.Pharmacological research in recent years shows, Rhizoma Polygonati Odorati has the effects such as enhancing immunity, atherosclerosis, adjusting myocardial movement frequency and intensity, blood fat reducing, slow down aging, antibiotic, blood vessel dilating, blood pressure lowering.
At present both at home and abroad research and the utilization of Rhizoma Polygonati Odorati are mainly contained to following several method: the one, Rhizoma Polygonati Odorati is dried to pulverizing, directly edible or add in the food such as flour edible; The 2nd, ion-exchange-resin process, the method is Rhizoma Polygonati Odorati purified water soaked overnight, spent ion exchange resin absorption Rhizoma Polygonati Odorati soak, then use the organic solvent eluting, and collect eluent and carry out distilling under reduced pressure, make the Rhizoma Polygonati Odorati concentrated solution; The 3rd, the organic solvent extraction method is extracted Rhizoma Polygonati Odorati in extractor by soak with ethanol, by after extracting liquid filtering, be placed in decompression concentrator and distilled, and obtains the Rhizoma Polygonati Odorati concentrated solution.The common problem of above-mentioned three kinds of methods is; technique is more extensive; technology is relatively backward; automaticity is low; the invalid impurity content of product is higher; the content of Rhizoma Polygonati Odorati effective ingredient lower (fragrant solomonseal rhizome polyoses content 3%～6%, Rhizoma Polygonati Odorati contents of steroid saponin 0.3%～0.8%), scale, standardized production are difficult to realize.
Summary of the invention
The objective of the invention is for overcoming the processing technique existed in prior art more extensive; technology is relatively backward; automaticity is low; the invalid impurity content of product is higher; the problems such as the Rhizoma Polygonati Odorati active constituent content is lower; and preparation method and the application thereof of a kind of high activity Rhizoma Polygonati Odorati concentrated solution that a kind of technical matters route is ripe, the product effective ingredient significantly improves, automaticity is higher of invention, the method is easy to realize scale, standardized production.
The present invention specifically provides a kind of preparation method of high activity Rhizoma Polygonati Odorati concentrated solution, it is characterized in that, comprises the steps:
(1) Rhizoma Polygonati Odorati enzymatic hydrolysis reaction: the Rhizoma Polygonati Odorati of cleaning is dropped into and forces in the outer circulation type extraction pot, the pure water that adds 6～12 times of Rhizoma Polygonati Odorati weight, utilize the natural biologic enzyme existed in the Rhizoma Polygonati Odorati plant, insulation under 25～45 ℃, enzymatic hydrolysis reaction 1～4h, the Rhizoma Polygonati Odorati enzyme is fully reacted with complexation materials such as albumen, starch fragrant solomonseal rhizome polyoses, steroidal saponin, discharge polysaccharide, saponin constituent, improve the effectively high activity component content such as fragrant solomonseal rhizome polyoses, Rhizoma Polygonati Odorati steroidal saponin;
(2) dynamic circulation is extracted: the Rhizoma Polygonati Odorati enzymatic hydrolysis reaction fully after, open the extraction pot circulating pump, extracting solution, by the heat exchanger fin dynamic heat that circulated, extracts 85～95 ℃ of temperature, extraction time 2～6h; Residue after filtration extracts once by above-mentioned technological parameter again, merges extracted twice liquid;
(3) micro-pore-film filtration: by said extracted liquid high-pressure delivery and pass through microporous membrane filters, discharge pressure is 0.3～0.6MPa, membrane filtration aperture 0.5～1.5 μ m, to remove solid particle polluter and larger molecular organics matter, then be transported to filtrate in temporary tank;
(4) concentrating under reduced pressure: open vacuum pump, make the concentrating under reduced pressure tank keep negative pressure state, the extracting solution that to keep in during be more than or equal to-0.03Mpa of vacuum in tank sucks in the concentrating under reduced pressure tank, opening steam valve is concentrated, 60～75 ℃ of thickening temperatures, vacuum-0.06～-0.08Mpa, be concentrated into solid content to 15%～25%Brix(w/w), be transported in temporary tank, cool the temperature to 4～12 ℃;
(5) high speed centrifugation: the feed liquid that will keep in tank is transported to macromolecular particles such as carrying out centrifugal, removal starch in centrifuge, rotating speed 12000～20000rpm, and separation factor 10000～15000,1～3h cleans a centrifuge, and centrifugal liquid is placed in temporary tank;
(6) secondary concentrating under reduced pressure: will keep in the low concentration concentrated solution after centrifugalize in tank and suck in the concentrating under reduced pressure tank, carry out the secondary concentrating under reduced pressure, 60～75 ℃ of thickening temperatures, vacuum-0.06～-0.08Mpa, when the solid content of concentrated solution reaches 60%～65%Brix(w/w) time, the concentrated end.
The high activity Rhizoma Polygonati Odorati concentrated solution that adopts By Anthrone Sulphuric acid colorimetric determination the method for the invention to make, its fragrant solomonseal rhizome polyoses content is 8%～12%; Adopt thin layer-spectrophotometry, Rhizoma Polygonati Odorati contents of steroid saponin 1.0%～2.5%.
The optimum process method that the present invention also provides described high activity Rhizoma Polygonati Odorati concentrated solution to prepare, specific as follows: (1) Rhizoma Polygonati Odorati enzymatic hydrolysis reaction: that the Rhizoma Polygonati Odorati of cleaning is dropped into and forces in the outer circulation type extraction pot, the pure water that adds 10 times of Rhizoma Polygonati Odorati weight, utilize the natural biologic enzyme existed in the Rhizoma Polygonati Odorati plant, insulation under 35 ℃, enzymatic hydrolysis reaction 2.5h;
(2) dynamic circulation is extracted: the Rhizoma Polygonati Odorati enzymatic hydrolysis reaction fully after, open the extraction pot circulating pump, extracting solution, by the heat exchanger fin dynamic heat that circulated, extracts 92 ℃ of temperature, extraction time 5h; Residue after filtration extracts once by above-mentioned technological parameter again, merges extracted twice liquid;
(3) micro-pore-film filtration: by said extracted liquid high-pressure delivery and by microporous membrane filters, discharge pressure is 0.4MPa, and membrane filtration aperture 1.0 μ m are transported to filtrate in temporary tank;
(4) concentrating under reduced pressure: open vacuum pump, make the concentrating under reduced pressure tank keep negative pressure state, the extracting solution that to keep in during reach-0.03Mpa of vacuum in tank sucks in the concentrating under reduced pressure tank, opening steam valve is concentrated, 65 ℃ of thickening temperatures, vacuum-0.07Mpa, be concentrated into solid content to 20%Brix(w/w), be transported in temporary tank, cool the temperature to 8 ℃;
(5) high speed centrifugation: will keep in feed liquid in tank be transported in the high speed tube centrifuge, carry out centrifugal, rotating speed 16000rpm, separation factor 12000,1.5h cleans a centrifuge, centrifugal liquid is placed in temporary tank;
(6) secondary concentrating under reduced pressure: will keep in the low concentration concentrated solution after centrifugalize in tank and suck in the concentrating under reduced pressure tank, carry out the secondary concentrating under reduced pressure, 68 ℃ of thickening temperatures, vacuum-0.07Mpa, when the solid content of concentrated solution reaches 65%Brix(w/w) time, the concentrated end.
The preparation method of high activity Rhizoma Polygonati Odorati concentrated solution of the present invention is characterized in that: described Rhizoma Polygonati Odorati comprises closes Rhizoma Polygonati Odorati, Polygonatum Modoratum (Mill.) Druce and processed goods thereof.
Can be used for making the products such as blood sugar lowering medicine, enhancing immunity medicine and health food by the preparation-obtained high activity Rhizoma Polygonati Odorati of high activity Rhizoma Polygonati Odorati concentrated solution preparation method of the present invention concentrated solution.
In the preparation technology of high activity Rhizoma Polygonati Odorati concentrated solution of the present invention, creationary invention, Rhizoma Polygonati Odorati enzymatic hydrolysis reaction technology, dynamic circulation extractive technique, secondary concentration technology, microporous membrane filter techniques, high-speed centrifugation technology have been used, make processing technique more advanced, improved automatic controlling level, effectively improved the content of concentrated solution fragrant solomonseal rhizome polyoses and Rhizoma Polygonati Odorati steroidal saponin, increased the stable physical-chemical indexes of product, product quality is improved significantly.The present invention has optimized the preparation technology parameter of high activity Rhizoma Polygonati Odorati concentrated solution, study the Rhizoma Polygonati Odorati natural biologic enzyme to complex qualitative response temperature, times such as fragrant solomonseal rhizome polyoses, steroidal saponin and albumen, starch, discharged to greatest extent fragrant solomonseal rhizome polyoses, the contour active component of steroidal saponin; The pressure of microporous filter membrane and flow velocity thereof, the selection of high speed centrifuge separation factor, K cryogenic treatment are treated to the parameters such as centrifugal concentrating liquid, concentrated number of times and concentration control have carried out the optimization of system, obviously improve yield, effective active component content and the product quality of product, can be used for making the products such as the medicine of blood sugar lowering, enhancing immunity and health food.Technology flow process of the present invention can be widely used in Chinese crude drug deep processing industry, is conducive to improve the added value of Chinese crude drug, promotes Chinese medicine processing modernization development.
The specific embodiment
Embodiment 1
(1) Rhizoma Polygonati Odorati enzymatic hydrolysis reaction: selected, clean and the clean sheet 500kg of the pass Rhizoma Polygonati Odorati of weighing drops into and forces in outer circulation type extraction pot (being called for short extraction pot in embodiment), add pure water 3000kg, utilize the natural biologic enzyme existed in the Rhizoma Polygonati Odorati plant, insulation under 25 ℃, enzymatic hydrolysis reaction time 1h.
(2) dynamic circulation is extracted: the Rhizoma Polygonati Odorati enzymatic hydrolysis reaction fully after, open the extraction pot circulating pump, extracting solution, by the heat exchanger fin dynamic heat that circulated, extracts 85 ℃ of temperature, extraction time 2h; Residue after filtration extracts once by above-mentioned technological parameter again, merges extracted twice liquid;
(3) micro-pore-film filtration: by said extracted liquid high-pressure delivery and by microporous membrane filters, discharge pressure is 0.3MPa, and membrane filtration aperture 0.6 μ m, to remove solid particle polluter and larger molecular organics matter, then is transported to filtrate in temporary tank;
(4) concentrating under reduced pressure: open vacuum pump, make the concentrating under reduced pressure tank keep negative pressure state, the extracting solution that to keep in during reach-0.03Mpa of vacuum in tank sucks in the concentrating under reduced pressure tank, opening steam valve is concentrated, 60 ℃ of thickening temperatures, vacuum-0.06Mpa, be concentrated into solid content to 15%Brix(w/w), be transported in temporary tank, cool the temperature to 4 ℃;
(5) high speed centrifugation: will keep in feed liquid in tank and be transported to GQLB-105N type tube centrifuge (embodiment of the present invention all adopts this centrifuge, hereinafter to be referred as centrifuge) in carry out centrifugal, remove the macromolecular particles such as starch, rotating speed 12000rpm, separation factor 11000,1h cleans a centrifuge, and centrifugal liquid is placed in temporary tank;
(6) secondary concentrating under reduced pressure: will keep in the low concentration concentrated solution after centrifugalize in tank and suck in concentration tank, carry out the secondary concentrating under reduced pressure, 60 ℃ of thickening temperatures, vacuum-0.06Mpa, when the solid content of concentrated solution reaches 60%Brix(w/w) time, the concentrated end.
By the enforcement of this extraction process, obtain high activity Rhizoma Polygonati Odorati concentrated solution 305.5kg, solid content is 60%Brix(w/w), the product yield is 61.1%, and fragrant solomonseal rhizome polyoses content is 8.7%, and the Rhizoma Polygonati Odorati contents of steroid saponin is 1.1%.
Embodiment 2
(1) Rhizoma Polygonati Odorati enzymatic hydrolysis reaction: selected, clean and the clean sheet 500kg of the Polygonatum Modoratum (Mill.) Druce of weighing drops in extraction pot, add pure water 6000kg, utilize the natural biologic enzyme existed in the Rhizoma Polygonati Odorati plant, insulation under 45 ℃, enzymatic hydrolysis reaction time 4h.
(2) dynamic circulation is extracted: the Rhizoma Polygonati Odorati enzymatic hydrolysis reaction fully after, open the extraction pot circulating pump, extracting solution, by the heat exchanger fin dynamic heat that circulated, extracts 95 ℃ of temperature, extraction time 6h; Residue after filtration extracts once by above-mentioned technological parameter again, merges extracted twice liquid;
(3) micro-pore-film filtration: by said extracted liquid high-pressure delivery and by microporous membrane filters, discharge pressure is 0.5MPa, and membrane filtration aperture 1.5 μ m, to remove solid particle polluter and larger molecular organics matter, then are transported to filtrate in temporary tank;
(4) concentrating under reduced pressure: open vacuum pump, make the concentrating under reduced pressure tank keep negative pressure state, the extracting solution that to keep in during reach-0.03Mpa of vacuum in tank sucks in the concentrating under reduced pressure tank, opening steam valve is concentrated, 72 ℃ of thickening temperatures, vacuum-0.08Mpa, be concentrated into solid content to 25%Brix(w/w), be transported in temporary tank, cool the temperature to 12 ℃;
(5) high speed centrifugation: the feed liquid that will keep in tank is transported to macromolecular particles such as carrying out centrifugal, removal starch in centrifuge, rotating speed 19000rpm, and separation factor 14000,2.5h cleans a centrifuge, and centrifugal liquid is placed in temporary tank;
(6) secondary concentrating under reduced pressure: will keep in the low concentration concentrated solution after centrifugalize in tank and suck in concentration tank, carry out the secondary concentrating under reduced pressure, 72 ℃ of thickening temperatures, vacuum-0.08Mpa, when the solid content of concentrated solution reaches 62%Brix(w/w) time, the concentrated end.
By the enforcement of this extraction process, obtain high activity Rhizoma Polygonati Odorati concentrated solution 301.0kg, solid content is 62%Brix(w/w), the product yield is 60.2%, and fragrant solomonseal rhizome polyoses content is 9.1%, and the Rhizoma Polygonati Odorati contents of steroid saponin is 1.3%.
Embodiment 3
(1) Rhizoma Polygonati Odorati enzymatic hydrolysis reaction: selected, clean and the clean sheet 500kg of the pass Rhizoma Polygonati Odorati of weighing drops in extraction pot, add pure water 5000kg, utilize the natural biologic enzyme existed in the Rhizoma Polygonati Odorati plant, insulation under 35 ℃, enzymatic hydrolysis reaction time 2.5h.
(2) dynamic circulation is extracted: the Rhizoma Polygonati Odorati enzymatic hydrolysis reaction fully after, open the extraction pot circulating pump, extracting solution, by the heat exchanger fin dynamic heat that circulated, extracts 92 ℃ of temperature, extraction time 5h; Residue after filtration extracts once by above-mentioned technological parameter again, merges extracted twice liquid;
(3) micro-pore-film filtration: by said extracted liquid high-pressure delivery and by microporous membrane filters, discharge pressure is 0.4MPa, and membrane filtration aperture 1.0 μ m, to remove solid particle polluter and larger molecular organics matter, then are transported to filtrate in temporary tank;
(4) concentrating under reduced pressure: open vacuum pump, make the concentrating under reduced pressure tank keep negative pressure state, the extracting solution that to keep in during reach-0.03Mpa of vacuum in tank sucks in the concentrating under reduced pressure tank, opening steam valve is concentrated, 65 ℃ of thickening temperatures, vacuum-0.07Mpa, be concentrated into solid content to 20%Brix(w/w), be transported in temporary tank, cool the temperature to 8 ℃;
(5) high speed centrifugation: the feed liquid that will keep in tank is transported to macromolecular particles such as carrying out centrifugal, removal starch in centrifuge, rotating speed 16000rpm, and separation factor 12000,1.5h cleans a centrifuge, and centrifugal liquid is placed in temporary tank;
(6) secondary concentrating under reduced pressure: will keep in the low concentration concentrated solution after centrifugalize in tank and suck in concentration tank, carry out the secondary concentrating under reduced pressure, 68 ℃ of thickening temperatures, vacuum-0.07Mpa, when the solid content of concentrated solution reaches 65%Brix(w/w) time, the concentrated end.
By the enforcement of this extraction process, obtain high activity Rhizoma Polygonati Odorati concentrated solution 355.2kg, solid content is 65%Brix(w/w), the product yield is 71.04%, and fragrant solomonseal rhizome polyoses content is 12%, and the Rhizoma Polygonati Odorati contents of steroid saponin is 2.5%.
Comparative Examples
(1) selected, clean and the clean sheet 500kg of the pass Rhizoma Polygonati Odorati of weighing drops in extraction pot, add pure water 5000kg, open the extraction pot circulating pump, extracting solution, by the heat exchanger fin dynamic heat that circulated, extracts 92 ℃ of temperature, extraction time 5h; Residue after filtration extracts once by above-mentioned technological parameter again, merges extracted twice liquid;
(2) micro-pore-film filtration: by said extracted liquid high-pressure delivery and by microporous membrane filters, discharge pressure is 0.4MPa, and membrane filtration aperture 1.0 μ m, to remove solid particle polluter and larger molecular organics matter, then are transported to filtrate in temporary tank;
(3) concentrating under reduced pressure: open vacuum pump, make the concentrating under reduced pressure tank keep negative pressure state, the extracting solution that to keep in during reach-0.03Mpa of vacuum in tank sucks in the concentrating under reduced pressure tank, opening steam valve is concentrated, 65 ℃ of thickening temperatures, vacuum-0.07Mpa, be concentrated into solid content to 20%Brix(w/w), be transported in temporary tank, cool the temperature to 8 ℃;
(4) high speed centrifugation: the feed liquid that will keep in tank is transported to macromolecular particles such as carrying out centrifugal, removal starch in centrifuge, rotating speed 16000rpm, and separation factor 12000,1.5h cleans a centrifuge, and centrifugal liquid is placed in temporary tank;
(5) secondary concentrating under reduced pressure: will keep in the low concentration concentrated solution after centrifugalize in tank and suck in concentration tank, carry out the secondary concentrating under reduced pressure, 68 ℃ of thickening temperatures, vacuum-0.07Mpa, when the solid content of concentrated solution reaches 65%Brix(w/w) time, the concentrated end.
By the enforcement of this extraction process, obtain Rhizoma Polygonati Odorati concentrated solution 310.9kg, solid content is 61%Brix(w/w), the product yield is 62.8%, and fragrant solomonseal rhizome polyoses content is 3.8%, and the Rhizoma Polygonati Odorati contents of steroid saponin is 0.35%.
(1), the experimentation (following experiment all adopts the Rhizoma Polygonati Odorati cream of embodiment 3 preparations as reagent agent) of high activity Rhizoma Polygonati Odorati concentrated solution (being called for short Rhizoma Polygonati Odorati cream, lower same) hypoglycemic activity
Embodiment 4
The impact of Rhizoma Polygonati Odorati cream on normal mouse blood sugar:
1. materials and methods: choose 50 of normal Kunming mouses, body weight 23～25g, be divided at random 5 groups, every group 10 (male and female half and half), difference per os gavage Rhizoma Polygonati Odorati cream 62.5g/kg, 18.75g/kg, 6.25g/kg, positive control drug antidiabetic drug 2.5mg/kg and normal saline, each administration group successive administration 15d, last administration 24h posterior orbit is dehematized (getting the front fasting 12h of blood), uses By Phosphomolybdate Colorimetry blood glucose.
2. result: the results are shown in Table 1.
After table 1 Rhizoma Polygonati Odorati cream successive administration 15d on the impact of normal mouse blood sugar value
Compare * P<0.05, * * P<0.01 with the normal saline group.
Result shows, continuously after per os gavage Rhizoma Polygonati Odorati cream 15d, normal mouse is all had to obvious hypoglycemic activity, high, medium and low dosage group Rhizoma Polygonati Odorati cream with contrast normal saline group (high dose group P<0.01 of comparing that all there were significant differences, middle dosage group P<0.05, low dose group P<0.01), antidiabetic drug also has obvious hypoglycemic activity (P<0.01), high dose group Rhizoma Polygonati Odorati cream is similar to the blood sugar lowering level of antidiabetic drug group, other dosage group hypoglycemic effects are not as good as the antidiabetic drug group, but on statistical significance all without significant difference (P > 0.05).
Embodiment 5
The impact of Rhizoma Polygonati Odorati cream on alloxan induced hyperglycemia mice:
1. modeling: get the above Kunming mouse of fasting 24h, tail vein injection alloxan normal saline solution 75mg/kg (0.1ml/10g), in 30S, injection is complete, after injection, the 48h eye socket is got blood and is surveyed blood glucose, get the front fasting 12h of blood, adopt By Phosphomolybdate Colorimetry blood glucose, select blood glucose value to be tested higher than the mice of 11.0mmol/L.
2. materials and methods: get the satisfactory Kunming mouse of blood glucose value after 50 modelings, male and female half and half, body weight 25～28g, be divided at random 5 groups, every group 10, get 10 normal mouses, also male and female half and half, as Normal group, started per os gastric infusion Rhizoma Polygonati Odorati cream 62.5g/kg respectively from the same day of dividing into groups, 18.75g/kg, 6.25g/kg, antidiabetic drug 7.5mg/kg and normal saline, measure respectively each modeling group 2d, 7d, 14d, blood glucose value during 21d, each measure the blood glucose time and be 24h after the last administration, measure the front fasting 12h of blood glucose, eye socket is got blood, adopt the By Phosphomolybdate Colorimetry blood glucose value.
3. result: the results are shown in Table 2.
The impact (n=10) of table 2 Rhizoma Polygonati Odorati cream different dosing time on alloxan diabetes mouse blood sugar value
Compare * P<0.05, * * P<0.01 with the normal saline group.
Result shows, Rhizoma Polygonati Odorati cream needs the hyperglycemia mice that the long period administration could cause alloxan to demonstrate hypoglycemic effect.In the time of can finding out administration 7d from experimental result between each dosage group of Rhizoma Polygonati Odorati cream and simulation group without significant difference (P<0.05), during administration 14d high dose group with contrast the normal saline group and compare that there were significant differences (P<0.05), other dosage groups have blood sugar lowering performance, but on statistical significance without significant difference (P > 0.05); During 21d, there were significant differences (P<0.01) for high dose group, middle dosage group and model group, and low dose group also has blood sugar reducing function, but on statistical significance without significant difference (P > 0.05); Demonstrate hypoglycemic activity faster after the administration of antidiabetic drug group, after administration 21d, can make blood glucose remain on lower level, hypoglycemic activity is better than each dosage group of Rhizoma Polygonati Odorati.Model group has been compared extremely significant difference with normal group, and model group blood glucose all is greater than 11.1mmol/L, meets requirement of experiment.
(2) experimentation of Rhizoma Polygonati Odorati cream enhancing immunity effect
Experimental example 6
The impact of Rhizoma Polygonati Odorati cream administration on the mice enhancing immunity
1. materials and methods: choose 40 of normal Kunming mouses, body weight 23～25g, divide 4 groups at random, every group 10 (male and female half and half), difference per os gavage Rhizoma Polygonati Odorati cream 62.5g/kg, 18.75g/kg, 6.25g/kg, positive control is distilled water, and each administration group successive administration 30d measures and respectively organizes the mouse immune capacity index.
1.1 delayed allergy experimental technique and investigation index: sheep red blood cell (SRBC) is brought out mouse DTH (the sufficient sole of the foot thickens method), get Sanguis caprae seu ovis, with normal saline washing 3 times, and with the centrifugal 10min of centrifuge 2000rpm, every mice is through lumbar injection Mianyang erythrocyte (SRBC) 0.2ml, after sensitization 4d, measures left back sufficient sole of the foot section thickness, same position is measured 3 times, averages.Overstock SRBC(v/v at measuring point subcutaneous injection 20%) 20ml, after injection, 24h measures left back sufficient sole of the foot section thickness, means the degree of DTH with the difference of sufficient sole of the foot thickness before and after injection SRBC.
1.2 antibody-producting cell detects (Jerne slide glass method): by hematocrit SRBC with being mixed with 2%(v/v) cell suspending liquid, every mouse peritoneal is injected 0.2ml, carries out immunity.After 4d, mice cervical vertebra dislocation is put to death, takes out spleen, use Hank ' s liquid washing 2 times, then by cell suspension in 5ml RPMI1640 culture fluid, count cell number, cell concentration is adjusted into to 5 * 10
6individual/ml.After the top layer culture medium is dissolved, 45 ℃ of water bath heat preservations, mix with Hank ' the s liquid of pH7.2～2 times of concentration of 7.4 equivalent, the every pipe 0.5ml of packing, then add 50 μ l 10%SRBC(v/v in pipe) and 25 μ l splenocyte suspensions (5 * 10
6individual/ml), mix homogeneously, be placed in the agarose slide, is placed in CO
2after incubation 1.5h, add SA buffer dilution complement (1:6) in incubator, continue incubation 1.5h, calculate the hemolysis plaque number.
1.3 Turnover of Mouse Peritoneal Macrophages is engulfed chicken red blood cell test (in vitro method): before experiment, 4d injects 2% hematocrit sheep red blood cell (SRBC) 0.2ml to every mouse peritoneal.Put to death mice with the cervical vertebra dislocation method, only give Hank ' s liquid 5ml/ that mice contains 5% calf serum through lumbar injection, extract ascites 1～2ml after 2min, the ascites that Hank ' s liquid is mixed with to 1% chicken RBC suspension 0.5ml and 0.5ml adds in vitro, mixes, and gets suspension 0.5ml smear, 37 ℃ of insulation 25min, after taking out, use normal saline from the slide backside rinse, fix, dye, read sheet, calculate phagocytic rate and phagocytic index.
2, result
2.1 the results are shown in Table 3:
The effect (n=10) of table 3 Rhizoma Polygonati Odorati cream administration to sheep red blood cell (SRBC) inducing mouse DTH
Compare * P<0.05, * * P<0.01 with the distilled water group.
The administration of Rhizoma Polygonati Odorati cream on the impact of sheep red blood cell (SRBC) (SRBC) inducing mouse DTH as can be seen from Table 3, the swelling degree of the paw of Rhizoma Polygonati Odorati cream high dose group, middle dosage group is higher than the distilled water matched group, difference all has significant (P<0.05, P<0.01), show that in this experiment, the administration of Rhizoma Polygonati Odorati cream can strengthen the delayed allergy of mice.
2.2 the results are shown in Table 4:
The impact (n=10) of table 4 Rhizoma Polygonati Odorati cream administration antagonist cellulation number
Compare * P<0.05, * * P<0.01 with the distilled water group.
The impact of Rhizoma Polygonati Odorati cream administration antagonist cellulation as can be seen from Table 4, Rhizoma Polygonati Odorati cream high dose group haemolysis speckle number/full spleen is higher than the solvent control group, difference has remarkable meaning (P<0.05), shows that Rhizoma Polygonati Odorati cream high dose group, middle dosage group have facilitation to the propagation of mouse antibodies cellulation.
2.3 the results are shown in Table 5:
The impact (n=10) of chicken red blood cell test is engulfed in the administration of table 5 Rhizoma Polygonati Odorati cream on Turnover of Mouse Peritoneal Macrophages
Compare * P<0.05, * * P<0.01 with the distilled water group.
The impact that the administration of Rhizoma Polygonati Odorati cream is engulfed chicken red blood cell test to Turnover of Mouse Peritoneal Macrophages as can be seen from Table 5, Rhizoma Polygonati Odorati cream is processed the high dose group mouse macrophage and is engulfed chicken red blood cell ability (phagocytic rate and phagocytic index) higher than the distilled water matched group, difference has significant (phagocytic rate P<0.05, phagocytic index P<0.01), shows that Rhizoma Polygonati Odorati cream high dose group, middle dosage group engulf the effect of being significantly increased of chicken red blood cell ability to mouse macrophage.
3. conclusion
The result of the test demonstration, Rhizoma Polygonati Odorati cream all has remarkable effect to delayed allergy, antibody-producting cell number, the macrophage phagocytic chicken red blood cell ability of Kunming mouse; Rhizoma Polygonati Odorati cream high dose group, middle dosage group, low dose group and contrast the distilled water group relatively, high dose group, middle dosage group performance significant difference (P<0.05, P<0.01).In sum, Rhizoma Polygonati Odorati cream has the effect of stronger enhancing human body immunity power.
Above-mentionedly experimental results show that Rhizoma Polygonati Odorati cream has blood sugar lowering and enhancing immunity effect.Comprehensive each result of the test, can tentatively think when the effective dosage of mice is 62.5g/kg, can reduce the blood sugar level of diabetic mice, strengthens the mice immunity of organisms.
Above-described embodiment is only explanation technical conceive of the present invention and characteristics, and its purpose is to allow the person skilled in the art can understand content of the present invention and implement according to this, can not limit the scope of the invention with this.All equivalences that spirit is done according to the present invention change or modify, within all should being encompassed in protection scope of the present invention.
Claims (4)
1. the preparation method of a high activity Rhizoma Polygonati Odorati concentrated solution, is characterized in that, comprises the steps:
(1) Rhizoma Polygonati Odorati enzymatic hydrolysis reaction: the Rhizoma Polygonati Odorati of cleaning is dropped into and forces, in the outer circulation type extraction pot, to add the pure water of 6～12 times of Rhizoma Polygonati Odorati weight, utilize the natural biologic enzyme existed in the Rhizoma Polygonati Odorati plant, insulation under 25～45 ℃, enzymatic hydrolysis reaction 1～4h;
(2) dynamic circulation is extracted: the Rhizoma Polygonati Odorati enzymatic hydrolysis reaction fully after, open the extraction pot circulating pump, extracting solution, by the heat exchanger fin dynamic heat that circulated, extracts 85～95 ℃ of temperature, extraction time 2～6h; Residue after filtration extracts once by above-mentioned technological parameter again, merges extracted twice liquid;
(3) micro-pore-film filtration: by said extracted liquid high-pressure delivery and by microporous membrane filters, discharge pressure is 0.3～0.6MPa, and membrane filtration aperture 0.5～1.5 μ m is transported to filtrate in temporary tank;
(4) concentrating under reduced pressure: open vacuum pump, make the concentrating under reduced pressure tank keep negative pressure state, the extracting solution that to keep in during be more than or equal to-0.03Mpa of vacuum in tank sucks in the concentrating under reduced pressure tank, opening steam valve is concentrated, 60～75 ℃ of thickening temperatures, vacuum-0.06～-0.08Mpa, be concentrated into solid content to 15%～25%Brix(w/w), be transported in temporary tank, cool the temperature to 4～12 ℃;
(5) high speed centrifugation: will keep in feed liquid in tank be transported in centrifuge, carry out centrifugal, rotating speed 12000～20000rpm, separation factor 10000～15000,1～3h cleans a centrifuge, centrifugal liquid is placed in temporary tank;
(6) secondary concentrating under reduced pressure: will keep in the low concentration concentrated solution after centrifugalize in tank and suck in the concentrating under reduced pressure tank, carry out the secondary concentrating under reduced pressure, 60～75 ℃ of thickening temperatures, vacuum-0.06～-0.08Mpa, when the solid content of concentrated solution reaches 60%～65%Brix(w/w) time, the concentrated end.
2. according to the preparation method of the described high activity Rhizoma Polygonati Odorati of claim 1 concentrated solution, it is characterized in that: described Rhizoma Polygonati Odorati comprises closes Rhizoma Polygonati Odorati, Polygonatum Modoratum (Mill.) Druce and processed goods thereof.
3. according to the preparation method of the described high activity Rhizoma Polygonati Odorati of claim 1 concentrated solution, it is characterized in that, concrete steps are:
(1) Rhizoma Polygonati Odorati enzymatic hydrolysis reaction: the Rhizoma Polygonati Odorati of cleaning is dropped into and forces, in the outer circulation type extraction pot, to add the pure water of 10 times of Rhizoma Polygonati Odorati weight, utilize the natural biologic enzyme existed in the Rhizoma Polygonati Odorati plant, insulation under 35 ℃, enzymatic hydrolysis reaction 2.5h;
(2) dynamic circulation is extracted: the Rhizoma Polygonati Odorati enzymatic hydrolysis reaction fully after, open the extraction pot circulating pump, extracting solution, by the heat exchanger fin dynamic heat that circulated, extracts 92 ℃ of temperature, extraction time 5h; Residue after filtration extracts once by above-mentioned technological parameter again, merges extracted twice liquid;
(3) micro-pore-film filtration: by said extracted liquid high-pressure delivery and by microporous membrane filters, discharge pressure is 0.4MPa, and membrane filtration aperture 1.0 μ m are transported to filtrate in temporary tank;
(4) concentrating under reduced pressure: open vacuum pump, make the concentrating under reduced pressure tank keep negative pressure state, the extracting solution that to keep in during reach-0.03Mpa of vacuum in tank sucks in the concentrating under reduced pressure tank, opening steam valve is concentrated, 65 ℃ of thickening temperatures, vacuum-0.07Mpa, be concentrated into solid content to 20%Brix(w/w), be transported in temporary tank, cool the temperature to 8 ℃;
(5) high speed centrifugation: will keep in feed liquid in tank be transported in centrifuge, carry out centrifugal, rotating speed 16000rpm, separation factor 12000,1.5h cleans a centrifuge, centrifugal liquid is placed in temporary tank;
(6) secondary concentrating under reduced pressure: will keep in the low concentration concentrated solution after centrifugalize in tank and suck in the concentrating under reduced pressure tank, carry out the secondary concentrating under reduced pressure, 68 ℃ of thickening temperatures, vacuum-0.07Mpa, when the solid content of concentrated solution reaches 65%Brix(w/w) time, the concentrated end.
4. the application of the preparation method of the described high activity Rhizoma Polygonati Odorati of claim 1 concentrated solution aspect making blood sugar lowering medicine, enhancing immunity medicine and health food.
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| CN104026706B (en) * | 2014-06-27 | 2015-10-14 | 广西健美乐食品有限公司 | A kind of preparation method of red bamboo liquid |
| CN109959749A (en) * | 2017-12-26 | 2019-07-02 | 九芝堂股份有限公司 | A method of radix polygonati officinalis cream is identified using thin-layered chromatography |
| CN110917294A (en) * | 2019-12-10 | 2020-03-27 | 李学龙 | Preparation method of high-activity rhizoma polygonati cream |
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| CN110917294A (en) * | 2019-12-10 | 2020-03-27 | 李学龙 | Preparation method of high-activity rhizoma polygonati cream |
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