CN110343192B - Extraction method of huperzia serrata polysaccharide - Google Patents
Extraction method of huperzia serrata polysaccharide Download PDFInfo
- Publication number
- CN110343192B CN110343192B CN201910800020.XA CN201910800020A CN110343192B CN 110343192 B CN110343192 B CN 110343192B CN 201910800020 A CN201910800020 A CN 201910800020A CN 110343192 B CN110343192 B CN 110343192B
- Authority
- CN
- China
- Prior art keywords
- huperzia serrata
- solution
- polysaccharide
- huperzia
- drying
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Sustainable Development (AREA)
- Biochemistry (AREA)
- Materials Engineering (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicinal Chemistry (AREA)
- Polymers & Plastics (AREA)
- Organic Chemistry (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses a method for extracting huperzia serrata polysaccharide, and relates to the technical field of food and medicine processing. The extraction method of the huperzia serrata polysaccharide comprises the following steps: drying Huperzia serrata, pulverizing, and sieving to obtain Huperzia serrata powder; mixing the huperzia serrata powder with water and then extracting to obtain a huperzia serrata extract; decolorizing the huperzia serrata extract by macroporous resin and removing protein to obtain purified liquid; and carrying out alcohol precipitation treatment on the purified solution to obtain a precipitate, and drying to obtain the huperzia serrata polysaccharide. The extraction method of the huperzia serrata polysaccharide provided by the invention has the advantages of simple process and low cost, and the obtained huperzia serrata polysaccharide can effectively remove free radicals and plays a positive role in enhancing the immunity of an organism.
Description
Technical Field
The invention relates to the technical field of food and medicine processing, in particular to a method for extracting huperzia serrata polysaccharide.
Background
Huperzia serrata (Lycopodium serratum Thunb.) is Tibetan plant of huperzia of huperziaceae, also called Huperzia serrata, etc. Huperzia serrata is used as a traditional Chinese herbal medicine for treating traumatic injury, schizophrenia, inflammation and detoxification and the like. The Huperzia serrata contains Huperzia serrata polysaccharide, huperzine, terpenoids and flavonoids. In addition, the huperzia serrata contains more huperzia serrata polysaccharides, and has important effects of removing free radicals in human bodies, enhancing immunity and resisting oxidation.
At present, the research on the extraction of the huperzia serrata polysaccharide is not available, and the research on the extraction method of the huperzia serrata polysaccharide has important significance.
Disclosure of Invention
The invention mainly aims to provide an extraction method of huperzia serrata polysaccharide, and aims to solve the technical problem that huperzia serrata polysaccharide is difficult to extract.
In order to achieve the above object, the present invention provides a method for extracting huperzia serrata polysaccharide, comprising the steps of:
drying Huperzia serrata, pulverizing, and sieving to obtain Huperzia serrata powder;
mixing the huperzia serrata powder with water and then extracting to obtain a huperzia serrata extract;
decolorizing the huperzia serrata extract by macroporous resin and removing protein to obtain purified liquid;
and carrying out alcohol precipitation treatment on the purified solution to obtain a precipitate, and drying to obtain the huperzia serrata polysaccharide.
Optionally, in the step of drying, pulverizing and sieving the huperzia serrata to obtain the huperzia serrata powder, the drying conditions are as follows: the drying temperature is 60-70 ℃, and the drying time is 5-6 h; and/or the presence of a gas in the gas,
and the sieving is to sieve the mixture by a sieve of 60-140 meshes.
Optionally, the extracting step of mixing the huperzia serrata powder with water to obtain a huperzia serrata extract comprises:
mixing the huperzia serrata powder with water, and heating for 2-4 hours under the water bath condition of 40-60 ℃ to obtain a crude extract;
centrifuging the crude extract at a rotating speed of 6000-8000 r/min for 5-10min, and taking supernatant and carrying out suction filtration to obtain the huperzia serrata extract.
Optionally, in the step of mixing the huperzia serrata powder with water, and heating the mixture for 2 to 4 hours under the water bath condition of 40 to 60 ℃ to obtain the crude extract, the ratio of the mass of the huperzia serrata powder to the volume of the water is 1: (10-30).
Optionally, the step of subjecting the huperzia serrata extract to decolorization and deproteinization treatment by using macroporous resin to obtain a purified solution comprises:
decoloring the huperzia serrata extracting solution for 10-30 min at the temperature of 25-30 ℃ through macroporous resin to obtain a decoloring solution, wherein the volume ratio of the huperzia serrata extracting solution to the macroporous resin is (1-4): 1;
and mixing the decolorized solution with the Sevage solution, and layering to obtain a purified solution.
Optionally, in the step of mixing the decolored solution with the Sevage solution and layering to obtain the purified solution, the volume ratio of the decolored solution to the Sevage solution is (1-3): 1.
optionally, in the step of mixing the decolorized solution with a Sevage solution and layering to obtain a purified solution, the Sevage solution is a mixed solution of chloroform and n-butanol, and the volume ratio of chloroform to n-butanol in the mixed solution is (3-4): 1.
optionally, the step of precipitating the purified solution with ethanol to obtain a precipitate and drying the precipitate to obtain huperzia serrata polysaccharide comprises:
adding an ethanol solution into the purified solution, and standing for 10-16 h at 3-5 ℃ to obtain a solid-liquid mixed solution;
and carrying out solid-liquid separation on the solid-liquid mixed solution to obtain a precipitate, and drying to obtain the huperzia serrata polysaccharide.
Optionally, in the step of adding an ethanol solution into the purification, and then standing the mixture at 3 to 5 ℃ for 10 to 16 hours to obtain a solid-liquid mixed solution, the volume ratio of the purified solution to the ethanol solution is 1: (1-3); and/or the presence of a gas in the gas,
the volume concentration of the ethanol solution is 60-90%.
Optionally, in the step of precipitating the purified solution with ethanol to obtain a precipitate and drying the precipitate to obtain huperzia serrata polysaccharide, the precipitate is dried by freeze drying under the following conditions: the drying temperature is-20 to-10 ℃, and the drying time is 36 to 48 hours.
The extraction method of the huperzia serrata polysaccharide provided by the invention comprises the steps of mixing huperzia serrata polysaccharide powder with water for extraction, decoloring by using macroporous resin, deproteinizing, precipitating by using alcohol to obtain huperzia serrata polysaccharide precipitate, and finally drying the huperzia serrata polysaccharide precipitate to obtain the huperzia serrata polysaccharide with higher purity. The extraction method of the huperzia serrata polysaccharide provided by the invention comprises the steps of dissolving the huperzia serrata polysaccharide in the huperzia serrata in water through extraction to form an extracting solution, purifying the extracting solution through decoloring and deproteinization, and finally carrying out alcohol precipitation to obtain a high-purity huperzia serrata polysaccharide precipitate. The process is simple, the cost is low, and the obtained huperzia serrata polysaccharide can effectively remove free radicals and plays a positive role in enhancing the immunity of the organism.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other related drawings can be obtained according to the drawings without creative efforts.
FIG. 1 is a schematic flow chart of an embodiment of the method for extracting Huperzia serrata polysaccharides according to the present invention;
FIG. 2 is a graph of a glucose standard curve in the determination of glucose content;
FIG. 3 is a graph showing the trend of the scavenging rate of the huperzia serrata polysaccharides prepared in examples 1, 2, 3 and 4 for ABTS radicals in the determination of antioxidant activity;
FIG. 4 measurement of antioxidant Activity of Huperzia serrata polysaccharides prepared in examples 1, 2, 3 and 4 versus Fe2+A trend graph of clearance change.
The implementation, functional features and advantages of the objects of the present invention will be further explained with reference to the accompanying drawings.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
Huperzia serrata is a Chinese herbal medicine with excellent performance, contains various substances beneficial to the body, including Huperzia serrata polysaccharide, and the extraction of the Huperzia serrata polysaccharide is not researched at present. In view of the above, the extraction method of huperzia serrata polysaccharide provided by the invention can obtain huperzia serrata polysaccharide with higher purity, the process is simple, the cost is low, and the obtained huperzia serrata polysaccharide can effectively remove free radicals and plays a positive role in enhancing the immunity of the organism.
Fig. 1 is a schematic flow chart of an embodiment of the method for extracting huperzia serrata polysaccharide according to the present invention, wherein the method for extracting huperzia serrata polysaccharide comprises the following steps:
and step S10, drying the huperzia serrata, crushing and sieving to obtain huperzia serrata powder.
According to the technical scheme, the huperzia serrata polysaccharide with high purity can be obtained by taking huperzia serrata as a raw material and carrying out a series of treatments. The picked huperzia serrata contains a large amount of water, and needs to be dried, the drying temperature is preferably 60-70 ℃, the drying time is preferably 5-6 hours, so that huperzia serrata polysaccharide in the huperzia serrata is protected from being damaged, and meanwhile, the water is fully evaporated. Wherein, the drying temperature can be 60 ℃, 62 ℃, 65 ℃, 67 ℃, 70 ℃ and the like, the drying time can be 5h, 5.2h, 5.5h, 5.6h, 5.8h, 6h and the like, and the preferable drying conditions are as follows: the drying temperature is 60 ℃, and the drying time is 6 h.
The dried and crushed huperzia serrata needs to be sieved, so that the particle size of the obtained huperzia serrata powder is uniform, extraction of huperzia serrata polysaccharide is facilitated, the sieving is preferably performed by a 60-140-mesh sieve which can be 60 meshes, 70 meshes, 80 meshes, 100 meshes, 120 meshes, 140 meshes and the like, and the sieving is preferably performed by a 140-mesh sieve, so that the particle size of the obtained huperzia serrata powder is small, the surface area is large, and the subsequent extraction of the huperzia serrata polysaccharide is facilitated.
And step S20, mixing the huperzia serrata powder with water and then extracting to obtain a huperzia serrata extract.
When the step S20 is specifically implemented, the method may include the following steps:
step S21, mixing the huperzia serrata powder with water, and heating for 2-4 hours under the condition of water bath at the temperature of 40-60 ℃ to obtain a crude extract;
and step S22, centrifuging the crude extract at the rotating speed of 6000-8000 r/min for 5-10min, and taking supernatant and carrying out suction filtration to obtain the huperzia serrata extract.
Wherein, when the step S21 is performed, the ratio of the mass of the huperzia serrata powder to the volume of water is 1: (10-30) in (g/ml), specifically, 10ml, 18ml, 20ml, 22ml, 25ml, 30ml and the like of water can be added for every 1g of the huperzia serrata powder, and preferably, 20ml of water is added for every 1g of the huperzia serrata powder, so that the huperzia serrata polysaccharide is fully extracted and has moderate concentration. In addition, ultrapure water is used as water in the present embodiment in order not to introduce other impurities in the preparation of the extraction liquid.
And step S30, decoloring the huperzia serrata extract by macroporous resin, and removing protein to obtain a purified solution.
When the step S30 is specifically implemented, the implementation includes the following two steps:
step S31, decoloring the huperzia serrata extract by macroporous resin at 25-30 ℃ for 10-30 min to obtain a decolored solution, wherein the volume ratio of the huperzia serrata extract to the macroporous resin is (1-4): 1;
and step S32, mixing the decolored solution and the Sevage solution, and layering to obtain a purified solution.
In step S31, the macroporous resin is selected from nonionic macroporous resin with the model of Amberlite XAD-4, and the decoloring effect is good.
In step S32, the volume ratio of the destaining solution to the Sevage solution is (1-3): 1, such as 1:1, 2:1, 2.5:1, 3:1 and the like, wherein the Sevage solution is a mixed solution of chloroform and n-butyl alcohol, and the volume ratio of the chloroform to the n-butyl alcohol is (3-4): 1. in addition, in order to save the Sevage solution for protein removal, the destaining solution obtained in step S31 may be concentrated first and then mixed with the Sevage solution, so that the removal rate of protein is improved and the raw material is also saved.
And step S40, carrying out alcohol precipitation on the purified solution to obtain a precipitate, and drying the precipitate to obtain the huperzia serrata polysaccharide.
Step S40 is also performed in two steps:
step S41, adding an ethanol solution into the purified solution, and then standing for 10-16 h at 3-5 ℃ to obtain a solid-liquid mixed solution;
and step S42, performing solid-liquid separation on the solid-liquid mixed solution to obtain a precipitate, and drying the precipitate to obtain the huperzia serrata polysaccharide.
Wherein, in the step of alcohol precipitation, the volume ratio of purification liquid and ethanol solution and the concentration of ethanol solution all can exert an influence to the effect of alcohol precipitation, and the volume ratio of purification liquid and ethanol solution is 1: (1-3); the volume concentration of the ethanol is 60-90%.
After the huperzia serrata polysaccharide is precipitated by alcohol, solid-liquid separation is carried out on a solid-liquid mixture, the solid huperzia serrata polysaccharide is dried, the drying mode can be freeze drying, the drying temperature is-20 to-10 ℃, the drying time is 36 to 48 hours, and the original chemical composition and physical properties of the huperzia serrata polysaccharide after drying can be kept.
The extraction method of the huperzia serrata polysaccharide provided by the invention comprises the steps of mixing huperzia serrata polysaccharide powder with water for extraction, decoloring by using macroporous resin, deproteinizing, precipitating by using alcohol to obtain huperzia serrata polysaccharide precipitate, and finally drying the huperzia serrata polysaccharide precipitate to obtain the huperzia serrata polysaccharide with higher purity. The extraction method of the huperzia serrata polysaccharide provided by the invention is simple in process and low in cost, and the obtained huperzia serrata polysaccharide can effectively remove free radicals and plays a positive role in enhancing the immunity of an organism. In addition, in the technical scheme provided by the invention, the extraction conditions (extraction temperature, ratio of the mass of the huperzia serrata to the volume of water, and extraction time) of the huperzia serrata polysaccharide are optimized, so that the extraction rate of the huperzia serrata polysaccharide is improved.
The technical solutions of the present invention are further described in detail below with reference to specific examples and drawings, it should be understood that the following examples are merely illustrative of the present invention and are not intended to limit the present invention.
Example 1
Drying Huperzia serrata at 60 deg.C for 6h, and sieving with 140 mesh sieve to obtain Huperzia serrata powder. Mixing the huperzia serrata powder with ultrapure water, wherein the ratio of the mass of the huperzia serrata powder to the volume of the ultrapure water is 1: 20, heating for 3h under the condition of 50 ℃ water bath to obtain crude extract, centrifuging the crude extract for 5min at the rotating speed of 8000r/min, taking supernatant, and performing suction filtration to obtain huperzia serrata extract. And then, decoloring the huperzia serrata extract by macroporous resin at the temperature of 30 ℃ for 30min to obtain a decolored solution, wherein the volume ratio of the huperzia serrata extract to the macroporous resin is 1:1, mixing a decolorized solution and a Sevage solution in a volume ratio of 1:1, mixing and layering to obtain a purified solution, wherein the volume ratio of chloroform to n-butanol in the Sevage solution is 3: 1. adding an ethanol solution with the volume fraction of 90% into the purified solution, wherein the volume ratio of the purified solution to the ethanol solution is 1: 3, standing at 4 ℃ for 12 hours to obtain a solid-liquid mixed solution, carrying out solid-liquid separation on the solid-liquid mixed solution to obtain a precipitate, and carrying out freeze drying at-20 ℃ for 48 hours to obtain the huperzia serrata polysaccharide.
Example 2
Drying Huperzia serrata at 70 deg.C for 5 hr, and sieving with 60 mesh sieve to obtain Huperzia serrata powder. Mixing the huperzia serrata powder with ultrapure water, wherein the ratio of the mass of the huperzia serrata powder to the volume of the ultrapure water is 1: 20, heating for 2 hours under the condition of water bath at 60 ℃ to obtain crude extract, centrifuging the crude extract for 10 minutes at the rotating speed of 6000r/min, taking supernatant, and carrying out suction filtration to obtain the huperzia serrata extract. And then, decoloring the huperzia serrata extract by macroporous resin at the temperature of 25 ℃ for 10min to obtain a decolored solution, wherein the volume ratio of the huperzia serrata extract to the macroporous resin is 4: 1, mixing a decolorized solution and a Sevage solution in a volume ratio of 3:1, mixing and layering to obtain a purified solution, wherein the volume ratio of chloroform to n-butanol in the Sevage solution is 4: 1. adding an ethanol solution with the volume fraction of 60% into the purified solution, wherein the volume ratio of the purified solution to the ethanol solution is 1:1, standing at 5 ℃ for 10 hours to obtain a solid-liquid mixed solution, carrying out solid-liquid separation on the solid-liquid mixed solution to obtain a precipitate, and carrying out freeze drying at-10 ℃ for 36 hours to obtain the huperzia serrata polysaccharide.
Example 3
Drying Huperzia serrata at 65 deg.C for 5.5h, and sieving with 100 mesh sieve to obtain Huperzia serrata powder. Mixing the huperzia serrata powder with ultrapure water, wherein the ratio of the mass of the huperzia serrata powder to the volume of the ultrapure water is 1: heating for 3h under 40 deg.C water bath condition to obtain crude extractive solution, centrifuging at 7000r/min for 8min, and vacuum filtering the supernatant to obtain herba Lycopodii Serrati extractive solution. And then, decoloring the huperzia serrata extract by macroporous resin at the temperature of 27 ℃ for 20min to obtain a decolored solution, wherein the volume ratio of the huperzia serrata extract to the macroporous resin is 3:1, mixing a decolorized solution and a Sevage solution in a volume ratio of 2:1, mixing and layering to obtain a purified solution, wherein the volume ratio of chloroform to n-butanol in the Sevage solution is 3.5: 1. adding an ethanol solution with the volume fraction of 70% into the purified solution, wherein the volume ratio of the purified solution to the ethanol solution is 1: 2, standing at 3 ℃ for 16h to obtain a solid-liquid mixed solution, carrying out solid-liquid separation on the solid-liquid mixed solution to obtain a precipitate, and freeze-drying at-15 ℃ for 42h to obtain the huperzia serrata polysaccharide.
Example 4
Drying Huperzia serrata at 62 deg.C for 6h, and sieving with 80 mesh sieve to obtain Huperzia serrata powder. Mixing the huperzia serrata powder with ultrapure water, wherein the ratio of the mass of the huperzia serrata powder to the volume of the ultrapure water is 1: 30, heating for 4 hours under the condition of water bath at 50 ℃ to obtain crude extract, centrifuging the crude extract for 6 minutes at the rotating speed of 7000r/min, taking supernatant, and carrying out suction filtration to obtain the huperzia serrata extract. And then, decoloring the huperzia serrata extract by macroporous resin at the temperature of 29 ℃ for 27min to obtain a decoloring solution, wherein the volume ratio of the huperzia serrata extract to the macroporous resin is 2:1, mixing a decolorized solution and a Sevage solution in a volume ratio of 1:1, mixing and layering to obtain a purified solution, wherein the volume ratio of chloroform to n-butanol in the Sevage solution is 3.7: 1. adding an ethanol solution with the volume fraction of 80% into the purified solution, wherein the volume ratio of the purified solution to the ethanol solution is 1: and 3, standing at 4 ℃ for 13 hours to obtain a solid-liquid mixed solution, performing solid-liquid separation on the solid-liquid mixed solution to obtain a precipitate, and performing freeze drying at-17 ℃ for 40 hours to obtain the huperzia serrata polysaccharide.
Example 5
Drying Huperzia serrata at 68 deg.C for 5h, and sieving with 120 mesh sieve to obtain Huperzia serrata powder. Mixing the huperzia serrata powder with ultrapure water, wherein the ratio of the mass of the huperzia serrata powder to the volume of the ultrapure water is 1: 20, heating for 3h under the condition of 50 ℃ water bath to obtain crude extract, centrifuging the crude extract for 9min at the rotating speed of 8000r/min, taking supernatant, and carrying out suction filtration to obtain the huperzia serrata extract. And then, decoloring the huperzia serrata extract by macroporous resin at the temperature of 30 ℃ for 25min to obtain a decolored solution, wherein the volume ratio of the huperzia serrata extract to the macroporous resin is 1:1, mixing a decolorized solution and a Sevage solution in a volume ratio of 3:1, mixing and layering to obtain a purified solution, wherein the volume ratio of chloroform to n-butanol in the Sevage solution is 3: 1. adding an ethanol solution with the volume fraction of 90% into the purified solution, wherein the volume ratio of the purified solution to the ethanol solution is 1:1, standing at 3 ℃ for 15 hours to obtain a solid-liquid mixed solution, carrying out solid-liquid separation on the solid-liquid mixed solution to obtain a precipitate, and carrying out freeze drying at-13 ℃ for 38 hours to obtain the huperzia serrata polysaccharide.
Example 6
Drying Huperzia serrata at 60 deg.C for 6h, and sieving with 140 mesh sieve to obtain Huperzia serrata powder. Mixing the huperzia serrata powder with ultrapure water, wherein the ratio of the mass of the huperzia serrata powder to the volume of the ultrapure water is 1: 20, heating for 3h under the condition of 50 ℃ water bath to obtain crude extract, centrifuging the crude extract for 5min at the rotating speed of 8000r/min, taking supernatant, and performing suction filtration to obtain huperzia serrata extract. And then, decoloring the huperzia serrata extract by macroporous resin at the temperature of 25 ℃ for 15min to obtain a decolored solution, wherein the volume ratio of the huperzia serrata extract to the macroporous resin is 4: 1, mixing a decolorized solution and a Sevage solution in a volume ratio of 1:1, mixing and layering to obtain a purified solution, wherein the volume ratio of chloroform to n-butanol in the Sevage solution is 4: 1. adding an ethanol solution with the volume fraction of 90% into the purified solution, wherein the volume ratio of the purified solution to the ethanol solution is 1: 3, standing at 4 ℃ for 12h to obtain a solid-liquid mixed solution, carrying out solid-liquid separation on the solid-liquid mixed solution to obtain a precipitate, and carrying out freeze drying at-20 ℃ for 46h to obtain the huperzia serrata polysaccharide.
Comparative example 1
The procedure was as in example 1 except that the heating temperature of the water bath was 80 ℃.
Comparative example 2
The procedure was as in example 1 except that the heating time of the water bath was 1 hour.
Comparative example 3
The ratio of the mass of the huperzia serrata powder to the volume of the ultrapure water is 1: the other steps are the same as in example 1 except for 50.
The huperzia serrata polysaccharides prepared in the above examples and comparative examples were examined as follows.
Measurement of extraction ratio of polysaccharide from Huperzia serrata (first)
The huperzia serrata polysaccharides prepared in examples 1 to 6 and comparative examples 1 to 3 were measured as follows:
and (3) measuring a glucose content standard curve: weighing 0.01g of glucose, placing the glucose into a 100mL volumetric flask, adding distilled water to dissolve the glucose, diluting the glucose to a scale, and shaking up to obtain a glucose standard solution. 1.6mL, 3.2mL, 4.8mL, 6.4mL, 8mL, 12.8mL, 16mL of the glucose standard solution was pipetted into a 25mL volumetric flask, 2.5mL of a 5% phenol solution and 5mL of concentrated sulfuric acid were added, followed by shaking, water bath in a boiling water bath for 15min, and cooling to room temperature. The absorbance was measured at 490nm, and the glucose standard curve was plotted on the abscissa and the ordinate as the concentration of huperzine serrate polysaccharide, respectively, as shown in FIG. 2.
The method for measuring the extraction rate of the huperzia serrata polysaccharide comprises the following specific steps: dissolving the extracted huperzia serrata polysaccharide with distilled water, fixing the volume to 100mL, measuring the absorbance according to the glucose absorbance measuring method, obtaining the extraction rate of the huperzia serrata polysaccharide, and recording the extraction rate in table 1, wherein the extraction rate of the huperzia serrata polysaccharide is the mass of the extracted huperzia serrata polysaccharide divided by the mass of the huperzia serrata.
TABLE 1 comparison of Huperzia serrata polysaccharide extraction rates
As can be seen from table 1 above, the extraction rate of the huperzia serrata polysaccharide extracted by the extraction method provided by the present invention generally reaches more than 1.3%, which indicates that the method for extracting huperzia serrata polysaccharide provided by the present invention has significant advantages. Meanwhile, the extraction rate of the huperzia serrata polysaccharide in each proportion is generally lower than that of the embodiment, and the better advantage of the huperzia serrata polysaccharide prepared by the extraction method after the optimization of each parameter is also demonstrated.
(II) measurement of antioxidant Activity
(1) The scavenging properties of the huperzia serrata polysaccharides prepared in examples 1, 2, 3 and 4 for ABTS free radicals were determined as follows
Adding 20mL of potassium persulfate solution into 100mL of ABTS solution, reacting at room temperature in the dark for 24h, diluting with distilled water until the absorbance value at 734nm is about 0.7 to obtain a solution, placing the solution in the dark, repeating the above steps for 4 times to obtain 4 groups of solutions, adding 400 μ L of each huperzine serrate polysaccharide solution with the concentration of 1, 2, 4, 8mg/mL to the 4 groups of solutions to obtain 4 groups of samples, measuring the absorbance at 734nm of the samples respectively, and recording the absorbance as A1. Distilled water was used as a blank. The clearance was calculated according to the following formula and the clearance change was plotted as shown in fig. 3.
Clearance rate ═ 1- (A)1－A2)/A0]×100％。
In the formula A0Absorbance of control group; a. the1Is the absorbance of the sample; a. the2The background absorbance value of the sample (distilled water instead of ABTS + stock solution).
(2) The Huperzia serrata polysaccharides prepared in examples 1, 2, 3 and 4 were respectively tested for Fe as follows2+Scavenging properties of free radicals
Taking 100 mu L of each huperzia serrata polysaccharide solution with the concentration of 1, 2, 4 and 8mg/mL into test tubes, respectively adding 10 mu L of 5mmol/L ferrous chloride solution, 20 mu L of phenanthroline and 270mL of distilled water into each test tube, shaking uniformly to obtain 4 groups of samples, respectively measuring the absorbance of the samples at 562nm, and recording the absorbance as A1. Distilled water was used as a blank. The clearance was calculated according to the following formula and the clearance change was plotted as shown in fig. 4.
Clearance rate ═ 1- (A)1－A2)/A0]×100％。
In the formula A0Absorbance of control group; a. the1Is the absorbance of the sample; a. the2The background absorbance value of the sample (distilled water instead of ferrous chloride).
As can be seen from FIGS. 3 and 4, huperzia serrata polysaccharides are responsible for ABTS free radicals and Fe2+All have the function of removing and increasing the concentration of the huperzia serrata polysaccharide, and can be used for treating ABTS free radicals and Fe2+The clearance rate basically shows an increasing trend, which shows that the huperzia serrata polysaccharide prepared by the preparation method has better antioxidant activity.
In conclusion, the huperzia serrata polysaccharide prepared by the embodiment of the invention has better antioxidant activity, and can effectively remove ABTS free radicals and Fe2+The method can be widely used for preparing products with antioxidant effect and enhanced immunocompetence, and has wide application prospect.
The above is only a preferred embodiment of the present invention, and it is not intended to limit the scope of the invention, and various modifications and changes will occur to those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention shall be included in the scope of the present invention.
Claims (10)
1. The extraction method of the huperzia serrata polysaccharide is characterized by comprising the following steps of:
drying Huperzia serrata, pulverizing, and sieving to obtain Huperzia serrata powder;
mixing the huperzia serrata powder with water and then extracting to obtain a huperzia serrata extract;
decolorizing the huperzia serrata extract by macroporous resin and removing protein to obtain purified liquid;
and carrying out alcohol precipitation treatment on the purified solution to obtain a precipitate, and drying to obtain the huperzia serrata polysaccharide.
2. The method for extracting huperzia serrata polysaccharide according to claim 1, wherein the drying step comprises the steps of drying huperzia serrata, pulverizing and sieving to obtain huperzia serrata powder, wherein the drying conditions are as follows: the drying temperature is 60-70 ℃, and the drying time is 5-6 h; and/or the presence of a gas in the gas,
and the sieving is to sieve the mixture by a sieve of 60-140 meshes.
3. The method of claim 1, wherein the step of extracting the huperzia serrata extract comprises mixing the huperzia serrata powder with water to obtain a huperzia serrata extract solution, the step of extracting the huperzia serrata extract solution comprising:
mixing the huperzia serrata powder with water, and heating for 2-4 hours under the water bath condition of 40-60 ℃ to obtain a crude extract;
centrifuging the crude extract at a rotating speed of 6000-8000 r/min for 5-10min, and taking supernatant and carrying out suction filtration to obtain the huperzia serrata extract.
4. The method for extracting Huperzia serrata polysaccharides of claim 3, wherein in the step of mixing the Huperzia serrata powder with water, heating the mixture in a water bath at 40-60 ℃ for 2-4 hours to obtain the crude extract, the ratio of the mass of the Huperzia serrata powder to the volume of the water is 1 g: (10-30) ml.
5. The method for extracting huperzia serrata polysaccharide of claim 1, wherein the step of decolorizing the huperzia serrata extract through macroporous resin and deproteinizing the huperzia serrata extract to obtain a purified solution comprises:
decoloring the huperzia serrata extracting solution for 10-30 min at the temperature of 25-30 ℃ through macroporous resin to obtain a decoloring solution, wherein the volume ratio of the huperzia serrata extracting solution to the macroporous resin is (1-4): 1;
and mixing the decolorized solution with the Sevage solution, and layering to obtain a purified solution.
6. The method for extracting huperzia serrata polysaccharide according to claim 5, wherein in the step of mixing the decolored solution and the Sevage solution and obtaining the purified solution after layering, the volume ratio of the decolored solution to the Sevage solution is (1-3): 1.
7. the extraction method of huperzia serrata polysaccharide as claimed in claim 5, wherein in the step of mixing the decolorized solution with a Sevage solution and layering to obtain the purified solution, the Sevage solution is a mixed solution of chloroform and n-butanol, and the volume ratio of chloroform to n-butanol in the mixed solution is (3-4): 1.
8. the method for extracting huperzia serrata polysaccharide according to claim 1, wherein the step of precipitating the purified solution with ethanol to obtain a precipitate and drying the precipitate to obtain huperzia serrata polysaccharide comprises:
adding an ethanol solution into the purified solution, and standing for 10-16 h at 3-5 ℃ to obtain a solid-liquid mixed solution;
and carrying out solid-liquid separation on the solid-liquid mixed solution to obtain a precipitate, and drying to obtain the huperzia serrata polysaccharide.
9. The method for extracting huperzia serrata polysaccharide according to claim 8, wherein in the step of adding an ethanol solution to the purified solution and then standing the mixture at 3-5 ℃ for 10-16 hours to obtain a solid-liquid mixed solution, the volume ratio of the purified solution to the ethanol solution is 1: (1-3); and/or the presence of a gas in the gas,
the volume concentration of the ethanol solution is 60-90%.
10. The method for extracting huperzia serrata polysaccharide according to claim 1, wherein the step of precipitating the purified solution with ethanol to obtain a precipitate and drying the precipitate to obtain huperzia serrata polysaccharide, wherein the precipitate is dried by freeze drying under the following conditions: the drying temperature is-20 to-10 ℃, and the drying time is 36 to 48 hours.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910800020.XA CN110343192B (en) | 2019-08-27 | 2019-08-27 | Extraction method of huperzia serrata polysaccharide |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910800020.XA CN110343192B (en) | 2019-08-27 | 2019-08-27 | Extraction method of huperzia serrata polysaccharide |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110343192A CN110343192A (en) | 2019-10-18 |
| CN110343192B true CN110343192B (en) | 2021-04-27 |
Family
ID=68181288
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910800020.XA Active CN110343192B (en) | 2019-08-27 | 2019-08-27 | Extraction method of huperzia serrata polysaccharide |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110343192B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102048949A (en) * | 2009-11-05 | 2011-05-11 | 天津太平洋制药有限公司 | Use of lycopodium clavatum in preparation of medicament for treating bone diseases |
| CN102423321A (en) * | 2011-12-10 | 2012-04-25 | 重庆市秀山红星中药材开发有限公司 | Method for extracting general flavone from filter residue of huperzia serrata |
| CN105412613A (en) * | 2015-12-15 | 2016-03-23 | 湖北民族学院 | Composition for Alzheimer's disease and processing method of composition |
-
2019
- 2019-08-27 CN CN201910800020.XA patent/CN110343192B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102048949A (en) * | 2009-11-05 | 2011-05-11 | 天津太平洋制药有限公司 | Use of lycopodium clavatum in preparation of medicament for treating bone diseases |
| CN102423321A (en) * | 2011-12-10 | 2012-04-25 | 重庆市秀山红星中药材开发有限公司 | Method for extracting general flavone from filter residue of huperzia serrata |
| CN105412613A (en) * | 2015-12-15 | 2016-03-23 | 湖北民族学院 | Composition for Alzheimer's disease and processing method of composition |
Non-Patent Citations (2)
| Title |
|---|
| "伸筋草多糖提取的研究";张烨等;《农业开发与装备》;20180928(第9期);第139-140页 * |
| "植物多糖对正常及肿瘤小鼠T和B淋巴细胞的影响";顾远锡等;《军事医学（军事医学科学院院刊）》;19861227;第10卷(第6期);第401-405页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110343192A (en) | 2019-10-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN112110979B (en) | Method for extracting tea saponin | |
| JP6473803B2 (en) | Extraction method of chlorogenic acid from Tochu leaves | |
| CN113801013B (en) | Production process for extracting shikimic acid and/or ginkgo polysaccharide from ginkgo leaves | |
| CN110343192B (en) | Extraction method of huperzia serrata polysaccharide | |
| CN111748103B (en) | Method for purifying humin in soil | |
| CN110679996B (en) | Tobacco leaf containing selenium-enriched polysaccharide | |
| CN112759660A (en) | Citron polysaccharide and extraction method and application thereof | |
| CN110898091A (en) | Ginseng extract and preparation method thereof | |
| CN112321658B (en) | Method for extracting anthocyanin in aronia melanocarpa fruit | |
| CN107213180B (en) | Separation and extraction method of notoginseng flavone | |
| CN112876671B (en) | Extraction and purification method of plant condensed tannin and application thereof | |
| CN113599405A (en) | Method for comprehensively extracting multiple effective components from acanthopanax sessiliflorus fruits and application thereof | |
| CN107936133B (en) | Evening primrose leaf polysaccharide and preparation method thereof | |
| CN110679997B (en) | Tobacco shred containing tobacco leaf polysaccharide and derived polysaccharide with high moisturizing property | |
| CN113582835A (en) | Method for extracting shikimic acid from ginkgo leaf waste residue and application | |
| CN110872334A (en) | Method for extracting tea saponin from camellia oleifera shells | |
| CN113956319B (en) | Method for extracting and refining ecdysterone | |
| CN111544494A (en) | HPEF extraction method of tea seed phenolic acid | |
| CN111777656B (en) | Method for extracting astragaloside from fresh astragalus | |
| CN114426478B (en) | High-content gallic acid and preparation method thereof | |
| CN113956303B (en) | Method for simultaneously extracting salidroside, rosavin and polysaccharide from rhodiola rosea | |
| CN113912749B (en) | Method for removing polysaccharide pigment of gentiana straminea by using three-dimensional porous amino graphene decolorizing column | |
| CN114891052B (en) | Method for preparing 1,2,3, 6-tetra-O-galloyl-beta-D-glucopyranoside from camellia oleifera | |
| CN113694091B (en) | Method for extracting achyranthes bidentata chrysanthemum flavone | |
| CN115737708A (en) | Low-cost tartary buckwheat flavone preliminary rapid extraction and purification method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |