CN110194809A - The preparation method and application of Codonopsis pilosula polysaccharide iron complexes - Google Patents
The preparation method and application of Codonopsis pilosula polysaccharide iron complexes Download PDFInfo
- Publication number
- CN110194809A CN110194809A CN201910530623.2A CN201910530623A CN110194809A CN 110194809 A CN110194809 A CN 110194809A CN 201910530623 A CN201910530623 A CN 201910530623A CN 110194809 A CN110194809 A CN 110194809A
- Authority
- CN
- China
- Prior art keywords
- codonopsis pilosula
- pilosula polysaccharide
- iron complexes
- preparation
- solution
- 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.)
- Granted
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/02—Nutrients, e.g. vitamins, minerals
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
-
- 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
- C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Organic Chemistry (AREA)
- Veterinary Medicine (AREA)
- General Chemical & Material Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Public Health (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biochemistry (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Toxicology (AREA)
- Materials Engineering (AREA)
- Polymers & Plastics (AREA)
- Nutrition Science (AREA)
- Diabetes (AREA)
- Hematology (AREA)
- Obesity (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicinal Preparation (AREA)
Abstract
The present invention discloses a kind of preparation method and application of Codonopsis pilosula polysaccharide iron complexes, is related to macromolecule polysaccharide technical field.The preparation methods of the Codonopsis pilosula polysaccharide iron complexes is stirred evenly, forms mixed solution the following steps are included: citric acid is added in the aqueous solution of Codonopsis pilosula polysaccharide；Under agitation, Xiang Suoshu mixed solution and dripping liquor ferri trichloridi and sodium hydroxide solution react to obtain mixture；By the mixture separation and purification treatment, Codonopsis pilosula polysaccharide iron complexes are obtained.Codonopsis pilosula polysaccharide iron complexes produced by the present invention are easy to be absorbed by organisms utilization, have high bioavilability, will not generate ill-effect to gastrointestinal tract, safe and non-toxic ferro element can be supplemented for body, and have preferable anti-tumor activity.
Description
Technical field
The present invention relates to macromolecule polysaccharide technical field, in particular to the preparation method of a kind of Codonopsis pilosula polysaccharide iron complexes and
Using.
Background technique
Iron is required microelement in human body, is the hemoglobin in blood of human body, the weight of myoglobins and a variety of enzymes
Want component part.When internal iron deficiency or when utilizing generation obstacle of iron, hemoglobin synthesis can be made to reduce, newborn red blood cell blood
Lactoferrin amount is insufficient；Anaemia can not only occur when too little iron, the shortage of internal iron content enzyme can also be caused, lead to cellular respiration
Obstacle occurs, influences the function of histoorgan, can clinically cause the dysfunction of the systems such as gastrointestinal tract, circulation, nerve.Iron lacks
Weary and hypoferric anemia is that the most common malnutrition and China micro nutrient deficit urgently to be resolved are asked in the world today
Topic.The oral absorption utilization rate of traditional iron-supplementing preparation is low, Yi Yinqi gastrointestinal side effect, and traditional iron-supplementing preparation course for the treatment of is long,
Patient is difficult to adhere to receiving whole-course treatment, to influence the compliance of patient medication.
Summary of the invention
The main object of the present invention is to propose a kind of preparation method and application of Codonopsis pilosula polysaccharide iron complexes, it is intended to be solved existing
Have that iron-supplementing preparation absorption rate is low, the defect more than side effect.
To achieve the above object, the present invention proposes a kind of preparation method of Codonopsis pilosula polysaccharide iron complexes, the Codonopsis pilosula polysaccharide
The preparation methods of iron complexes the following steps are included:
Citric acid is added in the aqueous solution of Codonopsis pilosula polysaccharide, is stirred evenly, mixed solution is formed；
Under agitation, Xiang Suoshu mixed solution and dripping liquor ferri trichloridi and sodium hydroxide solution react mixed
Close object；
By the mixture separation and purification treatment, Codonopsis pilosula polysaccharide iron complexes are obtained.
Optionally, described that citric acid is added in the aqueous solution of Codonopsis pilosula polysaccharide, it stirs evenly, forms the step of mixed solution
In rapid, the mass ratio of the Codonopsis pilosula polysaccharide contained in the aqueous solution of the citric acid and the Codonopsis pilosula polysaccharide is (2~4): 1.
Optionally, it is described under agitation, Xiang Suoshu mixed solution and dripping liquor ferri trichloridi and sodium hydroxide are molten
Liquid, react mixture the step of in, the concentration of the liquor ferri trichloridi is 1~2mol/L.
Optionally, it is described under agitation, Xiang Suoshu mixed solution and dripping liquor ferri trichloridi and sodium hydroxide are molten
Liquid, react mixture the step of in, in the sodium hydroxide solution mass fraction of sodium hydroxide be 10~25%.
Optionally, it is described under agitation, Xiang Suoshu mixed solution and dripping liquor ferri trichloridi and sodium hydroxide are molten
Liquid, react mixture the step of include:
Under agitation, Xiang Suoshu mixed solution and dripping liquor ferri trichloridi and sodium hydroxide solution are to form reaction
Liquid, and control rate of addition and make the pH 7~9 of the reaction solution；
To which after reaction, the reaction solution is placed under 60~80 DEG C of water baths, 1.5~2.5h is reacted, must be mixed
Object.
Optionally, described by the mixture separation and purification treatment, the step of obtaining Codonopsis pilosula polysaccharide iron complexes, includes:
The mixture is centrifuged 10~15min under 6000~8000r/min revolving speed, collects supernatant；
20~30h will be stood after the supernatant alcohol precipitation, takes precipitating and with ethanol washing, then in 6000~8000r/
It is centrifuged 10~15min under min revolving speed, collects precipitating vacuum drying, obtains Codonopsis pilosula polysaccharide iron complexes.
In addition, the invention also provides Radix Codonopsis made from a kind of preparation method of Codonopsis pilosula polysaccharide iron complexes as described above
Nu-Iron is preparing the application in iron-supplementing preparation.
In addition, the invention also provides Radix Codonopsis made from a kind of preparation method of Codonopsis pilosula polysaccharide iron complexes as described above
Nu-Iron is preparing the application in ovarian cancer resistance medicament.
In technical solution of the present invention, using Codonopsis pilosula polysaccharide as glycocide, with ferric chloride reaction, iron ion is made to load to Radix Codonopsis
On polysaccharide, due to Codonopsis pilosula polysaccharide be natural large biological molecule, degradable absorption, therefore, Codonopsis pilosula polysaccharide iron complexes obtained
It is easy to be absorbed by organisms utilization, there is high bioavilability, ill-effect will not be generated to gastrointestinal tract, can is body supplement peace
The ferro element of Atoxic；In addition to this, Codonopsis pilosula polysaccharide remove free radical, enhance human immunity, in terms of have
Apparent pharmacological action, can play positive effect to body, and and the combination of iron ion can also effectively enhance Codonopsis pilosula polysaccharide
Bioactivity.
Detailed description of the invention
In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show below
There is attached drawing needed in technical description to be briefly described, it should be apparent that, the accompanying drawings in the following description is only this
Some embodiments of invention for those of ordinary skill in the art without creative efforts, can be with
Other relevant attached drawings are obtained according to these attached drawings.
Fig. 1 is the flow diagram of an embodiment of Codonopsis pilosula polysaccharide iron complexes preparation method proposed by the present invention；
Fig. 2 is iron canonical plotting in iron content measurement；
Fig. 3 is Codonopsis pilosula polysaccharide iron complexes made from embodiment 1 in Antioxidative Activity Determination to the clearance rate of metal ion
Trend chart；
Fig. 4 is removing of the Codonopsis pilosula polysaccharide iron complexes to ABTS free radical made from embodiment 1 in Antioxidative Activity Determination
Rate trend chart；
Fig. 5 is in the cancer suppressing action detection of anti-tumor activity test at Codonopsis pilosula polysaccharide iron complexes made from embodiment 1
Manage the MTT testing result figure of ovarian cancer cell, wherein (a) is the cell electron microscope of Codonopsis pilosula polysaccharide iron complexes processing group, (b)
Cell activation assay figure after handling cell for Codonopsis pilosula polysaccharide iron complexes；
Fig. 6 is that Codonopsis pilosula polysaccharide iron made from embodiment 1 is multiple in cancer cell specific induction of apoptosis detection in anti-tumor activity test
Close the ROS testing result figure of object processing ovarian cancer cell；
Fig. 7 is that Codonopsis pilosula polysaccharide iron made from embodiment 1 is multiple in cancer cell specific induction of apoptosis detection in anti-tumor activity test
Close the Fluorescence Intensity Assays result figure of the ROS detection of object processing ovarian cancer cell；
Fig. 8 is that Codonopsis pilosula polysaccharide iron made from embodiment 1 is multiple in cancer cell specific induction of apoptosis detection in anti-tumor activity test
The mitochondrial membrane potential for closing object processing ovarian cancer cell detects fluorogram；
Fig. 9 is that Codonopsis pilosula polysaccharide iron made from embodiment 1 is multiple in cancer cell specific induction of apoptosis detection in anti-tumor activity test
The mitochondrial membrane potential for closing object processing ovarian cancer cell detects fluorometric result figure；
Figure 10 is that Codonopsis pilosula polysaccharide iron made from embodiment 1 is multiple in cancer cell specific induction of apoptosis detection in anti-tumor activity test
Close Immunofluorescence test γ H2AX and the RAD51 fluorogram of object processing ovarian cancer cell.
The embodiments will be further described with reference to the accompanying drawings for the realization, the function and the advantages of the object of the present invention.
Specific embodiment
It in order to make the object, technical scheme and advantages of the embodiment of the invention clearer, below will be in the embodiment of the present invention
Technical solution be clearly and completely described.The person that is not specified actual conditions in embodiment, according to normal conditions or manufacturer builds
The condition of view carries out.Reagents or instruments used without specified manufacturer is the conventional production that can be obtained by commercially available purchase
Product.
In addition, the meaning of the "and/or" occurred in full text, including three schemes arranged side by side, by taking " A and/or B " as an example, packet
The scheme for including A scheme or B scheme or A and B while meeting.
Asiderosis and hypoferric anemia are urgently to be resolved micro of the most common malnutrition and China in the world today
Nutrient lacks problem.The oral absorption utilization rate of traditional iron-supplementing preparation is low, Yi Yinqi gastrointestinal side effect, and tradition mends iron
The preparation course for the treatment of is long, and patient is difficult to adhere to receiving whole-course treatment, to influence the compliance of patient medication.
In consideration of it, the present invention proposes a kind of preparation method of Codonopsis pilosula polysaccharide iron complexes, a kind of Codonopsis pilosula polysaccharide can be prepared
Iron complexes, the Codonopsis pilosula polysaccharide iron complexes are easy to be absorbed and utilized, and benefit iron effect is good and will not cause gastrointestinal reaction.In conjunction with
The flow diagram of one embodiment of the Codonopsis pilosula polysaccharide iron complexes preparation method that Fig. 1 is proposed, the Codonopsis pilosula polysaccharide iron complexes
Preparation method the following steps are included:
Step S10, citric acid is added in the aqueous solution of Codonopsis pilosula polysaccharide, is stirred evenly, form mixed solution.
In technical solution of the present invention, using Codonopsis pilosula polysaccharide as ligand, be added citric acid into Codonopsis pilosula polysaccharide solution, then with trichlorine
Change iron reaction, loads to iron ion on Codonopsis pilosula polysaccharide.Wherein, the additional amount of citric acid influences whether the network of Codonopsis pilosula polysaccharide and iron
Reaction is closed, and then influences the iron-content of Codonopsis pilosula polysaccharide iron complexes, in the present embodiment, the citric acid and the Codonopsis pilosula polysaccharide
Aqueous solution in the mass ratio of Codonopsis pilosula polysaccharide that contains be (2~4): 1, i.e., mass ratio can for 2:1,2.1:1,2.6:1,3:1,
3.4:1,4:1 etc., preferably 3:1.Within this range, the higher Codonopsis pilosula polysaccharide iron complexes of iron-content can be obtained.In addition to this,
Sodium citrate, natrium hydrocitricum, trisodium citrate etc. are compared, the Codonopsis pilosula polysaccharide iron for the complex reaction that citric acid directly participates in is compound
Object has higher iron-content.
It should be noted that wherein, Codonopsis pilosula polysaccharide can buy acquisition in the market, can also voluntarily prepare.Its
Preparation method may is that clean Radix Codonopsis dry, pulverize after be sieved.Radix Codonopsis powder is placed in distilled water, in water bath with thermostatic control
Refluxing extraction in pot.It collects extracting solution and extracting solution is filtered by vacuum while hot, remove residue, collect filtrate.Filtrate is set
It is concentrated in revolving instrument, is then mixed in equal volume with chloroform-n-butanol (3:1, v/v) solution, stand liquid separation.Retain supernatant after
Continuous distillation, is subsequently placed in drying box and dries to get Codonopsis pilosula polysaccharide.The present invention does not limit the concentration of the aqueous solution of Codonopsis pilosula polysaccharide
System, only need the mass ratio of the quality of the Codonopsis pilosula polysaccharide contained in the aqueous solution of Codonopsis pilosula polysaccharide and the additional amount of citric acid be 1:(2~
4).
Step S20, under agitation, Xiang Suoshu mixed solution and dripping liquor ferri trichloridi and sodium hydroxide solution,
React to obtain mixture.
Wherein, the pH of the concentration of reactant, the temperature of reaction system and reaction system influence whether reaction efficiency and
Effect is complexed, through analysis of experiments, when the concentration of the liquor ferri trichloridi is 1~2mol/L, the temperature of reaction system be 60~
80 DEG C, the pH of reaction system is when being 7~9, the iron-content of Codonopsis pilosula polysaccharide iron complexes obtained is 10%~33.8% (w/w).
In addition, it is molten that the sodium hydroxide is added dropwise when the mass fraction of sodium hydroxide in sodium hydroxide solution is 10~25%
Liquid adjusts the pH of reaction system more quickly, precisely.
In addition, can specifically include following steps in implementation steps S20:
Step S21, under agitation, Xiang Suoshu mixed solution and dripping liquor ferri trichloridi and sodium hydroxide solution with
Reaction solution is formed, and control rate of addition to make the pH 7~9 of the reaction solution；
Step S22, to which after reaction, the reaction solution is placed under 60~80 DEG C of water baths, reaction 1.5~
2.5h obtains mixture.
Wherein, when carrying out step S21, to avoid the reactant concentration of regional area excessively high or local pH is beyond default
Range, liquor ferri trichloridi and sodium hydroxide solution are slowly added into mixed solution in a manner of being added dropwise, specific that speed is added dropwise
The degree present invention with no restriction, can be added dropwise with constant speed, can also be added dropwise with speed change, only the pH of reaction solution need to be made to be in 7~9 always
In range.During the reaction, when liquor ferri trichloridi and sodium hydroxide solution is initially added dropwise, will appear in reaction solution red
Brown precipitate and at once disappear, when red-brown precipitation no longer disappears, illustrate reaction terminate, stop be added dropwise liquor ferri trichloridi and
Sodium hydroxide solution.
Step S30, by the mixture separation and purification treatment, Codonopsis pilosula polysaccharide iron complexes are obtained.
When carrying out step S30, can be achieved by the steps of:
Step S31, the mixture is centrifuged to 10~15min under 6000~8000r/min revolving speed, collects supernatant；
Step S32,20~30h will be stood after the supernatant alcohol precipitation, take precipitating and with ethanol washing, then 6000~
It is centrifuged 10~15min under 8000r/min revolving speed, collects precipitating vacuum drying, obtains Codonopsis pilosula polysaccharide iron complexes.
Wherein exist, when carrying out alcohol precipitation, volume fraction can be used for 80~95% ethyl alcohol, additional amount is about supernatant
Three times of liquid product, after mixing the ethyl alcohol with supernatant, the volumetric concentration of ethyl alcohol is 70~76% in supernatant.
In technical solution of the present invention, using Codonopsis pilosula polysaccharide as ligand, with ferric chloride reaction, iron ion is set to load to Radix Codonopsis more
On sugar, since Codonopsis pilosula polysaccharide is natural large biological molecule, degradable absorption, therefore, Codonopsis pilosula polysaccharide iron complexes obtained are easy
In be absorbed by organisms utilize, have high bioavilability, and due to iron ion be after being complexed enter body, thus make iron from
Son will not be combined by other internal substances before being absorbed and influence to absorb, will not to gastrointestinal tract generation ill-effect, so as to
Think that body supplements safe and non-toxic ferro element；Meanwhile Codonopsis pilosula polysaccharide is being removed free radical, is being enhanced human immunity, anti-aging
Etc. there is apparent pharmacological action, can play positive effect to body, and and the combination of metal ion can also be effective
Enhance the bioactivity of Codonopsis pilosula polysaccharide.That is, Codonopsis pilosula polysaccharide iron complexes proposed by the present invention both can be used as safe nothing
Evil, the iron-supplementing preparation easily absorbed use, and can also be used as antioxidant use, can be also used for preparing ovarian cancer resistance medicament.
In addition to this, preparation method proposed by the present invention is to the selection of citric acid, reaction temperature, reaction pH and citric acid
Optimized with parameters such as the mass ratioes of Codonopsis pilosula polysaccharide, effectively increase the yield and iron-content of Codonopsis pilosula polysaccharide iron complexes,
Codonopsis pilosula polysaccharide iron complexes obtained are rich in ferro element, and iron content has reached 10~30%, and has high stability and water
Dissolubility is easy to be absorbed by organisms, and has excellent pharmacological action and bioactivity.
In addition, the invention also provides Radix Codonopsis made from a kind of preparation method of Codonopsis pilosula polysaccharide iron complexes as described above
Nu-Iron is preparing the application in iron-supplementing preparation.
Codonopsis pilosula polysaccharide iron complexes are rich in ferro element, and iron content has reached 10~33.8%, and has high stability
And water solubility, it is easy to be absorbed by organisms, there is excellent pharmacological action and bioactivity.While supplementing ferro element, may be used also
Effectively to enhance the immunocompetence etc. of body, it is a kind of high-quality iron-supplementing preparation, can be used directly as iron-supplementing preparation, it can also be with
As one of component, and other auxiliary materials (such as preservative, corrigent etc.), and/or with other components (such as other microelements
Replenishers etc.) iron-supplementing preparation use is made together.
In addition, the invention also provides Radix Codonopsis made from a kind of preparation method of Codonopsis pilosula polysaccharide iron complexes as described above
Nu-Iron is preparing the application in ovarian cancer resistance medicament.
Codonopsis pilosula polysaccharide iron complexes can inhibit human epithelial ovarian carcinoma cells proliferation and induce ovarian cellular apoptosis, therefore can make
For one of the component of ovarian cancer resistance medicament.In the specific implementation, the Codonopsis pilosula polysaccharide iron that above-mentioned preparation method can be prepared is compound
Object is added other drugs component and ovarian cancer resistance medicament is made in auxiliary material as one of main component.
Technical solution of the present invention is described in further detail below in conjunction with specific embodiments and the drawings, it should be understood that
Following embodiment is only used to explain the present invention, is not intended to limit the present invention.
Embodiment 1
Citric acid is added in the aqueous solution of Codonopsis pilosula polysaccharide, is stirred evenly, formed mixed solution, wherein citric acid with
The mass ratio of the Codonopsis pilosula polysaccharide contained in the aqueous solution of Codonopsis pilosula polysaccharide is 3:1.Under agitation, to mixed solution and dripping
Speed is added dropwise by control to form reaction solution in the sodium hydroxide solution that 2mol/L liquor ferri trichloridi and mass fraction are 20%
Degree makes the pH of reaction solution remain 9.When red-brown precipitation no longer disappears in reaction solution, stop that liquor ferri trichloridi and hydrogen is added dropwise
Sodium hydroxide solution simultaneously places reaction liquid under 60 DEG C of water baths, reacts 1.5h, obtains mixture.By mixture in 8000r/min
It is centrifuged 10min under revolving speed, collects supernatant；22h will be stood after supernatant alcohol precipitation, take precipitating and with ethanol washing, then existed
It is centrifuged 10min under 8000r/min revolving speed, collects precipitating vacuum drying, obtains Codonopsis pilosula polysaccharide iron complexes.
Embodiment 2
Citric acid is added in the aqueous solution of Codonopsis pilosula polysaccharide, is stirred evenly, formed mixed solution, wherein citric acid with
The mass ratio of the Codonopsis pilosula polysaccharide contained in the aqueous solution of Codonopsis pilosula polysaccharide is 3:1.Under agitation, to mixed solution and dripping
The sodium hydroxide solution that 1.8mol/L liquor ferri trichloridi and mass fraction are 25% is added dropwise with forming reaction solution by control
Speed makes the pH of reaction solution remain 8.When red-brown precipitation no longer disappears in reaction solution, stop be added dropwise liquor ferri trichloridi and
Sodium hydroxide solution simultaneously places reaction liquid under 80 DEG C of water baths, reacts 2h, obtains mixture.By mixture in 8000r/min
It is centrifuged 11min under revolving speed, collects supernatant；26h will be stood after supernatant alcohol precipitation, take precipitating and with ethanol washing, then existed
It is centrifuged 12min under 8000r/min revolving speed, collects precipitating vacuum drying, obtains Codonopsis pilosula polysaccharide iron complexes.
Embodiment 3
Citric acid is added in the aqueous solution of Codonopsis pilosula polysaccharide, is stirred evenly, formed mixed solution, wherein citric acid with
The mass ratio of the Codonopsis pilosula polysaccharide contained in the aqueous solution of Codonopsis pilosula polysaccharide is 3:1.Under agitation, to mixed solution and dripping
The sodium hydroxide solution that 1.5mol/L liquor ferri trichloridi and mass fraction are 18% is added dropwise with forming reaction solution by control
Speed makes the pH of reaction solution remain 8.When red-brown precipitation no longer disappears in reaction solution, stop be added dropwise liquor ferri trichloridi and
Sodium hydroxide solution simultaneously places reaction liquid under 70 DEG C of water baths, reacts 2.5h, obtains mixture.By mixture in 7000r/
It is centrifuged 12min under min revolving speed, collects supernatant；It will be stood for 24 hours after supernatant alcohol precipitation, take precipitating and with ethanol washing, then existed
It is centrifuged 13min under 7000r/min revolving speed, collects precipitating vacuum drying, obtains Codonopsis pilosula polysaccharide iron complexes.
Embodiment 4
Citric acid is added in the aqueous solution of Codonopsis pilosula polysaccharide, is stirred evenly, formed mixed solution, wherein citric acid with
The mass ratio of the Codonopsis pilosula polysaccharide contained in the aqueous solution of Codonopsis pilosula polysaccharide is 4:1.Under agitation, to mixed solution and dripping
The sodium hydroxide solution that 1.2mol/L liquor ferri trichloridi and mass fraction are 20% is added dropwise with forming reaction solution by control
Speed makes the pH of reaction solution remain 8.When red-brown precipitation no longer disappears in reaction solution, stop be added dropwise liquor ferri trichloridi and
Sodium hydroxide solution simultaneously places reaction liquid under 80 DEG C of water baths, reacts 2h, obtains mixture.By mixture in 6000r/min
It is centrifuged 13min under revolving speed, collects supernatant；20h will be stood after supernatant alcohol precipitation, take precipitating and with ethanol washing, then existed
It is centrifuged 10min under 8000r/min revolving speed, collects precipitating vacuum drying, obtains Codonopsis pilosula polysaccharide iron complexes.
Embodiment 5
Citric acid is added in the aqueous solution of Codonopsis pilosula polysaccharide, is stirred evenly, formed mixed solution, wherein citric acid with
The mass ratio of the Codonopsis pilosula polysaccharide contained in the aqueous solution of Codonopsis pilosula polysaccharide is 2:1.Under agitation, to mixed solution and dripping
Speed is added dropwise by control to form reaction solution in the sodium hydroxide solution that 1mol/L liquor ferri trichloridi and mass fraction are 10%
Degree makes the pH of reaction solution remain 7.When red-brown precipitation no longer disappears in reaction solution, stop that liquor ferri trichloridi and hydrogen is added dropwise
Sodium hydroxide solution simultaneously places reaction liquid under 80 DEG C of water baths, reacts 2.2h, obtains mixture.By mixture in 8000r/min
It is centrifuged 15min under revolving speed, collects supernatant；30h will be stood after supernatant alcohol precipitation, take precipitating and with ethanol washing, then existed
It is centrifuged 15min under 6000r/min revolving speed, collects precipitating vacuum drying, obtains Codonopsis pilosula polysaccharide iron complexes.
Comparative example 1
In addition to changing citric acid into trisodium citrate, other steps are same as Example 1.
Comparative example 2
In addition to the pH of reaction system is maintained at 9.5, other steps are same as Example 1.
Comparative example 3
In addition to the temperature of water-bath is changed to 90 DEG C, other steps are same as Example 1.
Comparative example 4
In addition to the mass ratio of the Codonopsis pilosula polysaccharide contained in the aqueous solution of citric acid and Codonopsis pilosula polysaccharide is changed to 1:1, other steps
It is rapid same as Example 1.
Codonopsis pilosula polysaccharide iron complexes prepared by the various embodiments described above and comparative example are detected as follows.
(1) iron content measures
Codonopsis pilosula polysaccharide iron complexes prepared by Example 1 to 5 and comparative example 1 to 4 measure as follows:
Iron content standard curve determination: taking the volumetric flask of 8 25m L, be separately added into iron standard solution 0,1,2,3,4,
5,6,7mL, then it is separately added into 10% ascorbic acid 1.75mL, 0.1% phenanthroline 2.5mL, distilled water constant volume shakes up, puts
10min is set, absorbance is measured at 510nm, using the concentration of iron as abscissa, absorbance is the standard song that ordinate draws iron
Line is shown in Fig. 2.
Iron content measures in Codonopsis pilosula polysaccharide iron complexes, specific steps are as follows: take dry Codonopsis pilosula polysaccharide iron complexes powder
0.05g is dissolved in appropriate distilled water, is shifted in 50mL volumetric flask and is settled to scale, and Codonopsis pilosula polysaccharide ferrous solution is made.Take Radix Codonopsis more
Sugared ferrous solution 1.0mL, is placed in 50mL volumetric flask, and 10% ascorbic acid solution 2.5mL, 0.1% phenanthroline solution is added
5.0mL is shaken up with distilled water constant volume, is reacted at 37 DEG C.Then absorbance is measured at 510nm, and iron is calculated according to the following formula
Content and be recorded in table 1.
Formula:
Wherein, Y is iron content (%), and A is absorbance.
The comparison of 1 iron content of table
By upper table 1 it is found that preparation method proposed by the present invention preparation Codonopsis pilosula polysaccharide iron complexes in iron ion content
Generally reach 17% or more, illustrates that Codonopsis pilosula polysaccharide iron complexes made from preparation method proposed by the present invention have aobvious in amount of iron load
Work advantage, and due to the pharmacological action of the easy absorbability of Codonopsis pilosula polysaccharide and itself, illustrate Codonopsis pilosula polysaccharide iron complexes as benefit iron
Agent not only has high bioavailability, also has effects that additionally positive.Meanwhile the iron content of each comparative example is generally lower than implementation
Example, also illustrating there is higher iron to contain Codonopsis pilosula polysaccharide iron complexes made from the preparation method of the present invention after each parameter optimization
Amount.
(2) Antioxidative Activity Determination
(1) Codonopsis pilosula polysaccharide iron complexes prepared by Example 1 measure it to metal cations Fe as follows2+Free radical
Cleaning performance:
Each 100 μ L of sample solution (1mg/mL, 2mg/mL, 4mg/mL, 8mg/mL, 10mg/mL) of various concentration is taken, respectively
10 μ L 5mmol/L FeCl are added2The distilled water of solution, 20 μ L phenanthrene hello piperazine solution and 270 μ L measures its extinction at 562nm
Degree is A1.Make blank control with distilled water.According to the following formula calculate clearance rate and draw clearance rate variation diagram as shown in figure 3,
Wherein CPPI is Codonopsis pilosula polysaccharide iron complexes.
Formula: clearance rate (%)=[1- (A1-A2)/A0] × 100%
In formula: A0For the absorbance of control group；A1For sample absorbance；A2For the background absorbance value (FeCl of sample2It is molten
Liquid changes distilled water into).
(2) Codonopsis pilosula polysaccharide iron complexes prepared by Example 1 measure its removing to ABTS free radical as follows
Performance:
Each 50 μ L of sample solution (1mg/mL, 2mg/mL, 4mg/mL, 8mg/mL, 10mg/mL) of various concentration is taken, respectively
200 μ L of ABTS+ stock solution is added, reacts 10min at room temperature, it is A1 that its absorbance is measured at 734nm.Make blank with distilled water
Control.Clearance rate is calculated according to the following formula and to draw clearance rate variation diagram as shown in Figure 4.
Formula: clearance rate (%)=[1- (A1-A2)/A0] × 100%
In formula: A0For the absorbance of control group；A1For sample absorbance；A2For the background absorbance value of sample, (distilled water is replaced
For ABTS+ stock solution).
Can be seen that Codonopsis pilosula polysaccharide iron complexes from Fig. 3, Fig. 4 has clearly metal ion free radical and ABTS free radical
Except effect, and with the raising of polysaccharide concentration of iron, the clearance rate of free radical is also increased, illustrates preparation method system of the present invention
The Codonopsis pilosula polysaccharide iron complexes obtained have preferable antioxidant activity.
(3) anti-tumor activity test
Codonopsis pilosula polysaccharide iron complexes prepared by Example 1 measure its anti-tumor activity as follows:
(1) cancer suppressing action
Experimental cell: ovarian cancer cell
Test method: will be inoculated in orifice plate after 0.25% trypsin digestion of the tumour cell of adherent growth, examination
It is compound to test group Codonopsis pilosula polysaccharide iron that each concentration (6.25 μ g/mL, 12.5 μ g/mL, 25 μ g/mL, 50 μ g/mL, 100 μ g/mL) is added
The aqueous solution of object, DEM/F12 culture medium is added in blank control group, in CO2After being cultivated for 24 hours in incubator, detected with mtt assay.It is real
Result is tested as shown in figure 5, abscissa is the concentration of Codonopsis pilosula polysaccharide iron complexes aqueous solution in Fig. 5 (b), ordinate is deposited for cell
Motility rate.
From figure 5 it can be seen that relative to blank control group, when the concentration of CPPI (Codonopsis pilosula polysaccharide iron complexes) is 6.25
When μ g/ml, 12.5 μ g/ml, 25 μ g/ml, 50 μ g/mL, cancer cell reduces 11.48% respectively, 12.49%, 16.8%,
17.23%；When its concentration is in 100 μ g/ml, it can lead to cancer cell Colony forming, substantially reduce 23.83% left and right
Cancer cell.Illustrate that Codonopsis pilosula polysaccharide iron complexes can produce dose dependent and reduce Colony forming ability, it can inhibit ovary
Cancer cell multiplication.
(2) cancer cell specific induction of apoptosis
Experimental cell: ovarian cancer cell
It is proceeded as follows according to the specification of kit: activation oxygen (ROS) detection, mitochondrial membrane potential (JC-1) inspection
It surveys, Nick End label (TUNEL) detection that deoxyribonucleotide terminal enzyme (DNA) mediates, cellular immunofluorescence detection.
Experiment sets 6 groups, and grouping situation is as follows:
First group: Codonopsis pilosula polysaccharide iron complexes are not added in blank control group；
The second~six group: Codonopsis pilosula polysaccharide iron complexes processing group, set respectively five concentration (6.25 μ g/mL, 12.5 μ g/mL,
25μg/mL、50μg/mL、100μg/mL)。
Test result is as shown in Fig. 6 to Figure 10.
As a result:
(1) abscissa is the concentration of Codonopsis pilosula polysaccharide iron complexes aqueous solution in Fig. 7, and ordinate is average fluorescent strength.From
In Fig. 6, Fig. 7 as can be seen that Codonopsis pilosula polysaccharide iron complexes processing group compared with the control group, the fluorescence intensity and ROS of DCF all compared with
It is high.
(2) in Fig. 8 merged represent fused cell, monomer represent JC-1 monomer aggregation in healthy cell formed it is poly-
Close object, aggregate represents JC-1 monomer in apoptosis or non-viable non-apoptotic cell；Abscissa is that Codonopsis pilosula polysaccharide iron complexes are water-soluble in Fig. 9
The concentration of liquid, ordinate are the ratio of apoptosis or non-viable non-apoptotic cell and healthy cell.From Fig. 8, Fig. 9 as can be seen that with Radix Codonopsis
The ratio of the increase of the concentration of Nu-Iron aqueous solution, apoptosis or non-viable non-apoptotic cell and healthy cell is gradually increasing, and illustrates party
Gracilis polysaccharide iron complexes can induce ovarian cellular apoptosis.
(3) it can be seen from fig. 10 that compared with blank control group, Codonopsis pilosula polysaccharide iron complexes processing group expresses γ H2AX
It is increased with RAD51 level and Codonopsis pilosula polysaccharide concentration of iron is positively correlated, illustrate that apoptotic cell gradually increases.
In conclusion Codonopsis pilosula polysaccharide iron complexes can inhibit human epithelial ovarian carcinoma cells proliferation and induce ovarian cellular apoptosis,
Illustrate that Codonopsis pilosula polysaccharide iron complexes prepared by the present invention have preferable anti-tumor activity.
The above is only a preferred embodiment of the present invention, is not intended to limit the scope of the invention, for this field
For technical staff, the invention may be variously modified and varied.All within the spirits and principles of the present invention, made any
Modification, equivalent replacement, improvement etc. should all be included within the scope of the present invention.
Claims (8)
1. a kind of preparation method of Codonopsis pilosula polysaccharide iron complexes, which comprises the following steps:
Citric acid is added in the aqueous solution of Codonopsis pilosula polysaccharide, is stirred evenly, mixed solution is formed；
Under agitation, Xiang Suoshu mixed solution and dripping liquor ferri trichloridi and sodium hydroxide solution react to obtain mixture；
By the mixture separation and purification treatment, Codonopsis pilosula polysaccharide iron complexes are obtained.
2. the preparation method of Codonopsis pilosula polysaccharide iron complexes as described in claim 1, which is characterized in that described that citric acid is added
In the step of into the aqueous solution of Codonopsis pilosula polysaccharide, stirring evenly, forming mixed solution, the citric acid and the Codonopsis pilosula polysaccharide
The mass ratio of the Codonopsis pilosula polysaccharide contained in aqueous solution is (2~4): 1.
3. the preparation method of Codonopsis pilosula polysaccharide iron complexes as described in claim 1, which is characterized in that described in stirring condition
Under, Xiang Suoshu mixed solution and dripping liquor ferri trichloridi and sodium hydroxide solution, react mixture the step of in, described three
The concentration of ferric chloride solution is 1~2mol/L.
4. the preparation method of Codonopsis pilosula polysaccharide iron complexes as described in claim 1, which is characterized in that described in stirring condition
Under, Xiang Suoshu mixed solution and dripping liquor ferri trichloridi and sodium hydroxide solution, react mixture the step of in, the hydrogen
The mass fraction of sodium hydroxide is 10~25% in sodium hydroxide solution.
5. the preparation method of Codonopsis pilosula polysaccharide iron complexes as described in claim 1, which is characterized in that described in stirring condition
Under, Xiang Suoshu mixed solution and dripping liquor ferri trichloridi and sodium hydroxide solution, react mixture the step of include:
Under agitation, Xiang Suoshu mixed solution and dripping liquor ferri trichloridi and sodium hydroxide solution to be to form reaction solution,
And it controls rate of addition and makes the pH 7~9 of the reaction solution；
To which after reaction, the reaction solution is placed under 60~80 DEG C of water baths, 1.5~2.5h is reacted, mixture is obtained.
6. the preparation method of Codonopsis pilosula polysaccharide iron complexes as described in claim 1, which is characterized in that described by the mixture
Separation and purification treatment, the step of obtaining Codonopsis pilosula polysaccharide iron complexes include:
The mixture is centrifuged 10~15min under 6000~8000r/min revolving speed, collects supernatant；
20~30h will be stood after the supernatant alcohol precipitation, takes precipitating and with ethanol washing, then at 6000~8000r/min turns
Speed is lower to be centrifuged 10~15min, collects precipitating vacuum drying, obtains Codonopsis pilosula polysaccharide iron complexes.
7. Codonopsis pilosula polysaccharide made from a kind of preparation method such as Codonopsis pilosula polysaccharide iron complexes as claimed in any one of claims 1 to 6
Iron complexes are preparing the application in iron-supplementing preparation.
8. Codonopsis pilosula polysaccharide made from a kind of preparation method such as Codonopsis pilosula polysaccharide iron complexes as claimed in any one of claims 1 to 6
Iron complexes are preparing the application in ovarian cancer resistance medicament.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910530623.2A CN110194809B (en) | 2019-06-18 | 2019-06-18 | Preparation method and application of codonopsis pilosula polysaccharide-iron compound |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910530623.2A CN110194809B (en) | 2019-06-18 | 2019-06-18 | Preparation method and application of codonopsis pilosula polysaccharide-iron compound |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110194809A true CN110194809A (en) | 2019-09-03 |
| CN110194809B CN110194809B (en) | 2021-10-26 |
Family
ID=67754801
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910530623.2A Active CN110194809B (en) | 2019-06-18 | 2019-06-18 | Preparation method and application of codonopsis pilosula polysaccharide-iron compound |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110194809B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115490775A (en) * | 2022-08-30 | 2022-12-20 | 广西民族师范学院 | Preparation method of sugar palm polysaccharide iron |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050147699A1 (en) * | 2003-12-31 | 2005-07-07 | Rey-Yuh Wu | Astragalus radix and codonopsis pilosulae radix mixed extract for inhibiting carcinogenesis and metastasis |
| CN101816697A (en) * | 2009-09-24 | 2010-09-01 | 兰州理工大学 | Method for preparing organic iron supplement by using radix codonopsitis |
| CN101967205A (en) * | 2010-10-12 | 2011-02-09 | 滨州学院 | Beta-dextriferron composite and preparation method thereof |
| CN102250261A (en) * | 2010-12-07 | 2011-11-23 | 天津怀仁制药有限公司 | New method for producing iron dextran |
| CN102850461A (en) * | 2011-06-28 | 2013-01-02 | 耿其尊 | Preparation process of corn polyferose |
| CN106478837A (en) * | 2016-11-26 | 2017-03-08 | 青岛海大生物集团有限公司 | A kind of can be used for green algae polysaccharide iron complexes of animals and plants iron supplement and preparation method thereof |
-
2019
- 2019-06-18 CN CN201910530623.2A patent/CN110194809B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050147699A1 (en) * | 2003-12-31 | 2005-07-07 | Rey-Yuh Wu | Astragalus radix and codonopsis pilosulae radix mixed extract for inhibiting carcinogenesis and metastasis |
| CN101816697A (en) * | 2009-09-24 | 2010-09-01 | 兰州理工大学 | Method for preparing organic iron supplement by using radix codonopsitis |
| CN101967205A (en) * | 2010-10-12 | 2011-02-09 | 滨州学院 | Beta-dextriferron composite and preparation method thereof |
| CN102250261A (en) * | 2010-12-07 | 2011-11-23 | 天津怀仁制药有限公司 | New method for producing iron dextran |
| CN102850461A (en) * | 2011-06-28 | 2013-01-02 | 耿其尊 | Preparation process of corn polyferose |
| CN106478837A (en) * | 2016-11-26 | 2017-03-08 | 青岛海大生物集团有限公司 | A kind of can be used for green algae polysaccharide iron complexes of animals and plants iron supplement and preparation method thereof |
Non-Patent Citations (4)
| Title |
|---|
| GE FENG ET AL.: ""Production of a codonopsis polysaccharide iron complex and evaluation of its properties"", 《INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES》 * |
| TAO XIN ET AL.: ""The inhibitory effect of a polysaccharide from Codonopsis pilosula on tumor growth and metastasis in vitro"", 《INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES》 * |
| 张新国等: ""党参多糖铁的合成及鉴别"", 《中医药学报》 * |
| 肖雄等: ""土党参多糖铁配合物的制备及其理化性质"", 《食品科学》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115490775A (en) * | 2022-08-30 | 2022-12-20 | 广西民族师范学院 | Preparation method of sugar palm polysaccharide iron |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110194809B (en) | 2021-10-26 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103265560B (en) | Application in gossypol/cotton ketone derivatives and preparation method thereof and antineoplastic | |
| CN110105460B (en) | Selenylation carboxymethyl pachyman and preparation method and application thereof | |
| CN109303922A (en) | A kind of Polysaccharides from Rosa roxburghii functionalized nano selenium compound and preparation method thereof and the application in hypoglycemic medicine | |
| CN111690150B (en) | Rare earth europium (III) coordination polymer with network structure and preparation method and application thereof | |
| Zhou et al. | A chiral responsive carbon dots–gold nanoparticle complex mediated by hydrogen peroxide independent of surface modification with chiral ligands | |
| Li et al. | Anion-exchangeable modulated fluorescence strategy for sensitive ascorbic acid detection with luminescent Eu hydroxy double salts nanosunflowers derived from MOFs | |
| CN110194809A (en) | The preparation method and application of Codonopsis pilosula polysaccharide iron complexes | |
| CN110016332B (en) | H+Modified red fluorescent gold nanocluster and preparation method and application thereof | |
| CN106290186A (en) | A kind of doping porous carbon Nano silver grain visualizing rapid mercury detection ion | |
| CN115960275A (en) | Preparation method of pachymaran iron with various pharmacological activities | |
| CN101481651B (en) | Metal nano material toxicity detection board and detection method thereof | |
| Sha et al. | One step functional assembly of guanosine monophosphate and terbium ion on metal organic frameworks for determination of alkaline phosphatase activity | |
| CN113632994A (en) | Method for improving soluble dietary fiber of roxburgh rose pomace | |
| CN108190849B (en) | Graphite phase carbon nitride nano particle and preparation method thereof | |
| CN114591509A (en) | Metal organic framework material with antioxidant activity | |
| CN109134683B (en) | Preparation method and application of pyracantha polysaccharide-iron complex | |
| CN113307892A (en) | Tremella polysaccharide iron (III) compound and preparation method thereof | |
| CN106006753B (en) | A kind of preparation method of Zn ferrite nanometer crystal assembly | |
| CN110051647A (en) | A kind of hericium erinaceum polysaccharide chelated zinc microcapsules and preparation method thereof | |
| CN110331098A (en) | A kind of sporogenic method of promotion blight dis-ease pathogen | |
| CN113667138B (en) | Silver-gold-metal organic framework compound surface enhanced Raman scattering substrate and preparation method and application thereof | |
| CN110419738A (en) | A kind of preparation method of lotus root polysaccharide selenium compound | |
| CN110818739B (en) | Metal iridium complex with synergistic response to tumor microenvironment pH/hypoxic and application thereof | |
| CN109429892A (en) | A kind of Phellinus selenium-rich bacterium powder | |
| CN114073316A (en) | Donkey spleen protein iron-supplementing oral liquid and preparation method thereof |
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 |