CN105713104A - Synthesis and application of selenized fenugreek polysaccharides - Google Patents
Synthesis and application of selenized fenugreek polysaccharides Download PDFInfo
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Abstract
The invention discloses a method for synthesizing selenized fenugreek polysaccharides. The method comprises the following steps: preparing a fenugreek polysaccharide formamide solution; preparing an ionic liquid; activating selenous acid by using the ionic liquid; and synthesizing the selenized fenugreek polysaccharides. The ionic liquid is utilized to activate the selenous acid to prepare the high-activity selenization reagent, and the bioactive selenized fenugreek polysaccharides are efficiently synthesized under mild conditions. The selenium content is up to 27554-29773 mu g/g. The reagent is friendly to the environment and simple to prepare, does not need any toxic seleniferous intermediate, and does not have the waste liquid discharge, thereby effectively shortening the reaction required lime, and widening the application range of the fenugreek polysaccharides as natural active substances. The activity test indicates that the synthesized selenized fenugreek polysaccharides have obvious in-vitro inhibiting actions on HepG2 liver cancer cells.
Description
Technical field
The invention belongs to biopolymer and chemosynthesis technical field, relate to the synthesis side of a kind of selenizing Fenugreek Polysaccharides
Method；The present invention also relates to the medical usage inhibitor application as HepG2 hepatoma carcinoma cell of selenizing Fenugreek Polysaccharides
In preparing in cancer therapy drug.
Background technology
Selenium (Se) is the requisite trace element of human body, life immune function raising, anticancer, antioxidation, prevent nutrition
All many-sides such as property hepatic necrosis play an important role.In biological metabolism, selenium is also the activity of glutathion peroxidase
Composition, therefore, selenium is particularly important to the effect of human body.But, selenium element is at earth skewness, in about five points of China
Three area lack selenium, and in bread and cheese, Se content is all extremely low, typically up to less than the effect of Selenium Supplement.At nature,
The existence form of selenium mainly includes inorganic selenium and organic selenium two kinds.Compared with inorganic selenium, organic selenium is safer, active higher
Containing selenium material, more more significantly than inorganic selenium on challenge.The main source of organic selenium is natural selenium-rich biology and people
Work synthesizes, and numerous rich in organic selenium material, selenium polysaccharide receives greatly concern.Selenium polysaccharide is as a kind of organic selenium chemical combination
Thing, has the activity of selenium and both polysaccharide.The biological activity of selenium polysaccharide is generally higher than polysaccharide and selenium, and toxicity is less, and after selenizing
Polysaccharide be easier to be absorbed by organism and utilize.
The preparation method of selenium polysaccharide includes natural containing selenium vegetable polysaccharides, microorganism enrichment culture metabolism selenium polysaccharide and artificial conjunction
Become selenium polysaccharide.Synthetic selenium polysaccharide is more convenient and controlled method, current domestic and foreign literature and patent mainly use with
Lower three kinds of methods prepare selenium polysaccharide: use monomer selenium, Monohydrated selenium dioxide or sodium selenite that polysaccharide is carried out selenizing modification under temperate condition；
Utilizing chemical property active, the intermediate with acyl chlorides structure is modified as selenizing reagent；Functional gene containing selenium is connect
Branch is on polysaccharide molecule.Liang Shuxuan etc. use the lower sodium selenate of glacial acetic acid catalysis to react with lycium barbarum polysaccharide, gained selenizing lycium barbarum polysaccharide
Se content at 606-3762 μ g/g(food research and development, 2011,3,160-164)；Li Zhizhou etc. utilize chemical synthesis,
Prepare polyporus selenium polysaccharide with polyporusum bellatus and sodium selenite for raw material, use continuously or ultrasonic assistant chemical synthesis process (food
Product and machinery, 2013,1,31-35, do not report Se content).Patent CN 1560088A has disclosed selenizing glucomannoglycan a kind of
Preparation method, selenium simple substance is oxidized to Se under oxidant effect6+, at Se6+The aqueous solution of ion adds ethanol and salt
Acid, obtains selenylation reaction liquid, is reacted with glucomannoglycan by selenylation reaction liquid, makes selenizing glucomannoglycan, gained selenizing Portugal
Sweet polysaccharide Se content ratio is relatively low, and for 125.4-197.3 μ g/g, and in derivant, selenium is Se6+；Patent ZL 88103347 disclosure
The preparation method of a kind of selenocarrageenan, with selenium powder as raw material, is dissolved with nitric acid and is prepared as selenium liquid, adds Kappa-chondrus ocellatus Holmes
Sol solution carries out selenylation reaction, but the method products therefrom Se content relatively low (2500-15000 ppm), and by-product Selenium hydride.
Poisonous.Patent CN102653566A discloses a kind of method that organic method prepares astragalus polysaccharide selenide, and the method is by astragalus polysaccharides
Crude product synthesizes sterling astragalus polysaccharides under enzyme degradation, then reacts with protochloride selenonyl, obtains the astragalus polysaccharides selenium content of selenizing
For 14305-24864 μ g/g, but the method selenizing time length (12-48h), reactant protochloride selenonyl is difficult to preparation, toxicity
Greatly, large-scale industrial production is not suitable for.Patent ZL 200910162003.4 discloses a kind of organic method and prepares selenizing Folium et Cacumen Artemisiae Halodendri
Polysaccharide or the method for fern amylose, react by fern amylose or artemisia desertorum polysaccharide and seleniyl chlorine to obtain the artemisia desertorum polysaccharide of selenizing or Radix potentillae anserinae
Polysaccharide, Se content 12000-26000 μ g/g.But the seleniyl chloro that the method uses becomes difficulty and seleniyl chlorine to have toxicity,
In course of reaction, exposing in atmosphere when adding seleniyl chlorine, seleniyl chlorine can be oxidized, may contain in the seleniyl chlorine obtained
There are the impurity such as selenonyl chlorine.
Generally there is the intermediate containing selenium and prepare complicated and malicious in the polysaccharide selenizing modification technique mentioned in above patent and document
Property big, response time length, selenizing polysaccharide Se content and the shortcoming such as yield is low, side reaction is many, biological effect is not clear.Ionic liquid
It is widely used in all kinds of organic synthesis with advantages such as its Heat stability is good, efficiency height, environmental friendliness and recyclable recyclings
In reaction.By consulting domestic and foreign literature and patent, all not reporting with Fenugreek Polysaccharides as raw material, ionic liquid is as catalyst
Synthesize method and the application of high Se content selenizing Fenugreek Polysaccharides.
Summary of the invention
It is an object of the invention to provide the synthetic method of a kind of selenizing Fenugreek Polysaccharides.
Another object of the present invention is to the inhibitor use providing a kind of selenizing Fenugreek Polysaccharides as HepG2 hepatoma carcinoma cell
In preparing cancer therapy drug.
One, the synthesis of selenizing Fenugreek Polysaccharides
1. the preparation of Fenugreek Polysaccharides formamide solution:
Fenugreek Polysaccharides stirs under the rotating speed of 300 ~ 500 rpm at 60 DEG C ~ 80 DEG C 2 ~ 5 h be dissolved in Methanamide, obtain calabash
The formamide solution of reed bar polysaccharide.
The weight average molecular weight of above-mentioned Fenugreek Polysaccharides is 2.303 × 106 Da, total sugar content is 87.2%.Fenugreek Polysaccharides with
The mass volume ratio of Methanamide is 2.5 ~ 4 mg/mL.
2. the preparation of ionic liquid
Concentrated sulphuric acid is added drop-wise in N-Methyl pyrrolidone with the speed of 50 ~ 80/min at-10 DEG C ~ 0 DEG C, 300 ~ 800
Rpm stirs reaction 10 ~ 18 h, washs 2 ~ 3 times by ethyl acetate after completion of the reaction, collect lower floor's thick liquid, vacuum under room temperature
It is dried to obtain ionic liquid.
Wherein: N-Methyl pyrrolidone and the body that volume ratio is 2:1 ~ 2.5:1, ethyl acetate and ionic liquid of concentrated sulphuric acid
Long-pending ratio is 0.5:1 ~ 1:1, and vacuum drying condition is vacuum 0.06 ~ 0.08 MPa, baking temperature 50 ~ 70 DEG C, time 6 ~ 12 h.
3. the ionic liquid activation to Monohydrated selenium dioxide
At 60 ~ 80 DEG C and N2Under conditions of protection, Monohydrated selenium dioxide is joined and above-mentioned ionic liquid reacts after 1 ~ 3 h must activate
Monohydrated selenium dioxide.
The mass volume ratio of above-mentioned Monohydrated selenium dioxide and ionic liquid is 0.125 ~ 0.2 g/mL.
4. the synthesis of selenizing Fenugreek Polysaccharides
With the speed of 30 ~ 60/min, the formamide solution Dropping funnel of Fenugreek Polysaccharides is added drop-wise to ionic liquid activate
Monohydrated selenium dioxide in, at 60 ~ 80 DEG C and N22 ~ 4 h are reacted, after question response under conditions of isolation air under conditions of protection
Add washing with acetone 10 ~ 15 times, precipitate is loaded in the bag filter of molecular cut off 8000 ~ 14000 Da flowing water dialysis 24 ~
48 h, deionized water dialysis 12 ~ 24 h, in complete back pkt. of dialysing, thing is evaporated to the 1/20 ~ 1/30 of original volume, at concentrated solution
Middle addition dehydrated alcohol (ethanol content of the total volume 70 ~ 80%) precipitates, centrifugal collection lower sediment thing, and lyophilization obtains selenium
Change Fenugreek Polysaccharides.
Wherein: the volume ratio of the formamide solution of Fenugreek Polysaccharides and the Monohydrated selenium dioxide of activation is 10 ~ 25, stirring condition is
The volume ratio of 300 ~ 800 rpm, acetone and reactant liquor is 2 ~ 3(v/v), concentrating under reduced pressure temperature is 55 ~ 65 DEG C, vacuum 0.06 ~
0.08 MPa, centrifugal rotational speed is 4000 ~ 5000 rpm/min, centrifugation time 10 ~ 15 min, and lyophilization condition is temperature-60
~-50 DEG C, vacuum 5 ~ 20 Pa, drying time 24 ~ 48 h.
Two, the external inhibitory action to HepG2 hepatoma carcinoma cell of selenizing Fenugreek Polysaccharides
Take the logarithm the HepG-2 cell of trophophase, use 0.25% trypsinization, and train with the DMEM containing 15% new-born calf serum
Concentration 3 × 10 made by foster base5The single cell suspension of individual/mL, constantly shake cell suspension makes cell number uniform, with 0.2 ml/ hole
It is inoculated in 96 well culture plates (edges of boards edge envelope sterilized water, blank group adds the 3 the most celliferous culture medium in hole), puts CO2In incubator
37 DEG C, saturated humidity, 5%CO2Under the conditions of cultivate.
Cultivating cell attachment after 24 h, change pastille culture medium, every hole adds 190 μ L culture medium and 10 μ L sample liquid are (blank
Group and matched group add 10 μ l PBS liquid), final concentration of 20,50,100,200,500,1000, the 5000 μ g/mL of medicine, every concentration
Repeat 6 holes, and set matched group and the blank zeroing group being not added with sample.Put CO2In incubator, 37 DEG C, saturated humidity, 5%CO2Bar
Cultivate under part.
After sample treatment 48 h, every hole adds MTT solution 20 μ L, continues to cultivate 4 h at 37 DEG C.Carefully discard in hole
Clear liquid, and add 150 μ L DMSO, vibrate 10 min, makes purple crystal be completely dissolved.At 490 nm, extinction is surveyed by microplate reader
Value, matched group returns to zero with blank group, and calculates suppression ratio by following equation, and calculates half according to the suppression ratio of variable concentrations sample
Number inhibition concentration (IC50):
Suppression ratio (%)=(1-experimental group mean OD value/matched group mean OD value) × 100%
IC to HepG2 hepatoma carcinoma cell50It is 527.4 ~ 1230.8 μ g/mL, there is the work of significant vitro inhibition HepG2 cell
With, can be used for developing cancer therapy drug as the inhibitor of HepG2 hepatoma carcinoma cell.
Beneficial effects of the present invention:
1, the present invention is with Fenugreek Polysaccharides as raw material, uses ionic liquid-catalyzed Monohydrated selenium dioxide to synthesize selenizing Fenugreek Polysaccharides, ion
Liquid is nontoxic, environmental protection, prepare simple, it is not necessary to use poisonous containing selenium intermediate, without discharging of waste liquid, be effectively shortened reaction required
Time, obtain the selenizing polysaccharide of high Se content under mild conditions.
2, it is prepared for the selenizing Fenugreek Polysaccharides with significant vitro inhibition HepG2 cytoactive, expands Semen Trigonellae
Polysaccharide is as the range of application of natural active matter；
The Se content of the selenizing Fenugreek Polysaccharides that 3, prepared by the present invention can be optimized by reaction condition regulation, with satisfied different necks
Territory requirement；
4, the present invention need not special installation, the most controlled, low cost, is suitable for popularization and application.
Accompanying drawing explanation
Fig. 1 Fenugreek Polysaccharides and the infrared spectrogram of selenizing Fenugreek Polysaccharides thereof.
X-ray photoelectron power spectrum (XPS) figure of Fig. 2 Fenugreek Polysaccharides and selenizing Fenugreek Polysaccharides.
Detailed description of the invention
Below by specific embodiment, the synthetic method of selenizing Fenugreek Polysaccharides of the present invention is further described.
Embodiment 1
(1) preparation of Fenugreek Polysaccharides formamide solution: by 200 mg Fenugreek Polysaccharides at 70 DEG C under rotating speed with 500 rpm
Stir 2 h to be dissolved in 60 mL Methanamides, obtain the formamide solution of Fenugreek Polysaccharides.
(2) preparation of ionic liquid: 5 mL concentrated sulphuric acids are added drop-wise to 10 mL's with the speed of 60/min at-10 DEG C
In N-Methyl pyrrolidone, 800 rpm stirring reaction 12 h, wash 2 times by 15 mL ethyl acetate under room temperature after completion of the reaction,
Collect lower floor thick liquid, at vacuum 0.07 MPa, be vacuum dried 6 h at 60 DEG C and obtain ionic liquid.
(3) ionic liquid activation to Monohydrated selenium dioxide: at 80 DEG C and N2Under conditions of protection, 0.6 g Monohydrated selenium dioxide is joined 4
ML ionic liquid reacts the Monohydrated selenium dioxide after 3 h must activate.
(4) synthesis of selenizing Fenugreek Polysaccharides: by the formamide solution Dropping funnel of 60 mL Fenugreek Polysaccharides with 40
Drip/speed of min is added drop-wise in the Monohydrated selenium dioxide that 4 mL ionic liquids activate, at 70 DEG C and N24 h are reacted under conditions of protection,
Under conditions of isolation air, add 128 mL acetone, washes at room temperature 10 times after question response, precipitate is loaded and retains
In the bag filter of molecular weight 8000 ~ 14000 Da, flowing water is dialysed 24 h, and deionized water is dialysed 18 h, and in complete back pkt. of dialysing, thing exists
0.08 MPa, being evaporated to the 1/30 of original volume at 60 DEG C, (ethanol content accounts for cumulative volume to add dehydrated alcohol in concentrated solution
70%) precipitation, under 5000 rpm, centrifugal 10 min collect lower sediment things, at 5 Pa, lyophilization 36 h at-55 DEG C, obtain selenium
Change Fenugreek Polysaccharides.
This selenizing Fenugreek Polysaccharides is kermesinus amorphous powder, total sugar content 80.3%, and in water, dissolubility is
18.5mg/mL(25 DEG C), Se content is 28128 μ g/g, and molecular weight is 2.376 × 106Da, the body to HepG2 hepatoma carcinoma cell
Outer IC50Value is 591.1 μ g/mL.
Below by Determination of Selenium, infrared spectrogram, X-ray photoelectron spectroscopic analysis (XPS), molecular weight determination to this
The structure of selenizing Fenugreek Polysaccharides of embodiment 1 synthesis, activity are analyzed, and are explained:
1. the mensuration of Se content: use fluorimetry that sample is carried out Determination of Selenium.
Weigh dry testing sample, add nitration mixture (HNO3:H2SO4:HClO4=1:1:4), at 135 DEG C after left undisturbed overnight
In clear up 1～2 hour, digestion solution is used distilled water diluting constant volume, take 10mL add 0.2 mol/L EDTA-2Na solution and
The oxammonium hydrochloride. each 2～4mL of 0.5 mol/L, stands 5～10min after shaken well, dark place adds the 2 of 0.1%, 3-diaminonaphthalene
Solution 2～4mL, boiling water bath 4～6min, takes out after being cooled to room temperature, adds hexamethylene 5 mL, abundant oscillation extraction, take hexamethylene
Alkane layer measures fluorescent value (excitation wavelength 376 nm launches wavelength 520 nm) in fluorescent sub-photometer, uses working curve method meter
Calculate Se content.
2. infrared spectrum map analysis
The infrared spectrogram of the selenizing Fenugreek Polysaccharides that Fig. 1 is Fenugreek Polysaccharides and prepared by the present invention.Fenugreek Polysaccharides in figure
Infrared spectrum shows that it has a characteristic peak of polysaccharide: 3400 cm-1The widest strong absworption peak in left and right is that the O-H of polysaccharide hydroxyl stretches
Vibration, 2922.22 cm-1Place is the stretching vibration of c h bond.Contrasted by the infrared spectrum with Fenugreek Polysaccharides, it can be seen that selenium
Change Fenugreek Polysaccharides is in addition to polysaccharide parent characteristic absorption peak, at 1149 cm-1Place and 669 cm-1There is new absworption peak in place,
It is respectively stretching vibration and the bending vibration of C-O-Se of Se=O, illustrates that the part-OH in Fenugreek Polysaccharides structure is by Monohydrated selenium dioxide
Base replaces, by selenizing.
3. XPS analysis
Fig. 2 is Fenugreek Polysaccharides and the XPS spectrum of selenizing Fenugreek Polysaccharides.The Se 3d spectrum of Fenugreek Polysaccharides does not has any peak, and
At 58.1 eV, selenizing Fenugreek Polysaccharides occurs in that the strongest Se 3d combines energy peak, the valence state of Se is Se4+, Fenugreek Polysaccharides is described
By selenizing, the selenium in selenizing Fenugreek Polysaccharides occurs with the form of monohydrate selenium dioxide ester.
4. molecular weight determination
Gel chromatography-dynamic static light scattering combination is used to measure, the Weight-average molecular of the selenizing Fenugreek Polysaccharides of preparation in the present invention
Amount is 1.533 × 106 Da ~2.616×106 Da。
Embodiment 2
(1) preparation of Fenugreek Polysaccharides formamide solution: by 250 mg Fenugreek Polysaccharides at 80 DEG C under rotating speed with 300 rpm
Stir 4 h to be dissolved in 100 mL Methanamides, obtain the formamide solution of Fenugreek Polysaccharides.
(2) preparation of ionic liquid: 4 mL concentrated sulphuric acids are added drop-wise to the speed of 50/min at 0 DEG C the N-of 10 mL
In methyl pyrrolidone, 600 rpm stirring reaction 18 h, wash 3 times by 7 mL ethyl acetate under room temperature after completion of the reaction, collect
Lower floor's thick liquid, at vacuum 0.06 MPa, is vacuum dried 8 h at 70 DEG C and obtains ionic liquid.
(3) ionic liquid activation to Monohydrated selenium dioxide: at 60 DEG C and N2Under conditions of protection, 0.8 g Monohydrated selenium dioxide is joined 4
ML ionic liquid reacts the Monohydrated selenium dioxide after 2 h must activate.
(4) synthesis of selenizing Fenugreek Polysaccharides: by the formamide solution Dropping funnel of 100 mL Fenugreek Polysaccharides with 60
Drip/speed of min is added drop-wise in the Monohydrated selenium dioxide that 4 mL ionic liquids activate, at 60 DEG C and N22 h are reacted under conditions of protection,
Under conditions of isolation air, add 312 mL acetone, washes at room temperature 12 times after question response, precipitate is loaded and retains
In the bag filter of molecular weight 8000 ~ 14000 Da, flowing water is dialysed 48 h, and deionized water is dialysed 12 h, and in complete back pkt. of dialysing, thing exists
0.06 MPa, being evaporated to the 1/25 of original volume at 65 DEG C, (ethanol content accounts for cumulative volume to add dehydrated alcohol in concentrated solution
80%) precipitation, under 4000 rpm, centrifugal 12 min collect lower sediment things, at 20 Pa, lyophilization 48 h at-60 DEG C,
Selenizing Fenugreek Polysaccharides.
This selenizing Fenugreek Polysaccharides is kermesinus amorphous powder, total sugar content 84.9%, and in water, dissolubility is 23.6
Mg/mL(25 DEG C), Se content is 27554 μ g/g, and molecular weight is 1.533 × 106Da, the external IC to HepG2 hepatoma carcinoma cell50
Value is 527.4 μ g/mL.
Embodiment 3
(1) preparation of Fenugreek Polysaccharides formamide solution: by 160 mg Fenugreek Polysaccharides at 60 DEG C under rotating speed with 400 rpm
Stir 5 h to be dissolved in 40 mL Methanamides, obtain the formamide solution of Fenugreek Polysaccharides.
(2) preparation of ionic liquid: 5 mL concentrated sulphuric acids are added drop-wise to the speed of 80/min at-5 DEG C the N-of 10 mL
In methyl pyrrolidone, 300 rpm stirring reaction 10 h, wash 3 times by 15 mL ethyl acetate under room temperature after completion of the reaction, receive
Collection lower floor thick liquid, at vacuum 0.08 MPa, is vacuum dried 12 h at 50 DEG C and obtains ionic liquid.
(3) ionic liquid activation to Monohydrated selenium dioxide: at 70 DEG C and N2Under conditions of protection, 0.6 g Monohydrated selenium dioxide is joined 4
ML ionic liquid reacts the Monohydrated selenium dioxide after 1 h must activate.
(4) synthesis of selenizing Fenugreek Polysaccharides: by the formamide solution Dropping funnel of 40 mL Fenugreek Polysaccharides with 30
Drip/speed of min is added drop-wise in the Monohydrated selenium dioxide that 4 mL ionic liquids activate, at 80 DEG C and N23 h are reacted under conditions of protection,
Under conditions of isolation air, add 110 mL acetone, washes at room temperature 15 times after question response, precipitate is loaded and retains
In the bag filter of molecular weight 8000 ~ 14000 Da, flowing water is dialysed 36 h, and deionized water is dialysed 24 h, and in complete back pkt. of dialysing, thing exists
0.07 MPa, being evaporated to the 1/20 of original volume at 55 DEG C, (ethanol content accounts for cumulative volume to add dehydrated alcohol in concentrated solution
75%) precipitation, under 4500 rpm, centrifugal 15 min collect lower sediment things, at 15 Pa, lyophilization 24 h at-50 DEG C,
Selenizing Fenugreek Polysaccharides.
This selenizing Fenugreek Polysaccharides is kermesinus amorphous powder, total sugar content 78.1%, and in water, dissolubility is 20.2
Mg/mL(25 DEG C), Se content is 29773 μ g/g, and molecular weight is 2.616 × 106Da, the external IC to HepG2 hepatoma carcinoma cell50
Value is 630.6 μ g/mL.
Comparative example 1
(1) preparation of Fenugreek Polysaccharides formamide solution: by 250 mg Fenugreek Polysaccharides at 80 DEG C under rotating speed with 300 rpm
Stir 4 h to be dissolved in 100 mL Methanamides, obtain the formamide solution of Fenugreek Polysaccharides.
(2) synthesis of selenizing Fenugreek Polysaccharides: 0.8 g Monohydrated selenium dioxide is joined the formamide solution of above-mentioned Fenugreek Polysaccharides
In, at 60 DEG C and N2React 2 h under conditions of protection, after question response, under conditions of isolation air, add 312 mL third
Ketone, washes at room temperature 12 times, precipitate is loaded flowing water in the bag filter of molecular cut off 8000 ~ 14000 Da and dialyses 48 h,
Deionized water is dialysed 12 h, and in complete back pkt. of dialysing, thing is evaporated to the 1/25 of original volume, dense at 0.06 MPa, at 65 DEG C
Adding dehydrated alcohol (ethanol content of the total volume 80%) precipitation in contracting liquid, under 4000 rpm, centrifugal 12 min collect lower floor and sink
Shallow lake thing, at 20 Pa, lyophilization 48 h at-60 DEG C, obtains selenizing Fenugreek Polysaccharides.
This selenizing Fenugreek Polysaccharides is pale red amorphous powder, total sugar content 82.8%, and in water, dissolubility is 16.6
Mg/mL(25 DEG C), Se content is 4060 μ g/g, and molecular weight is 1.236 × 106Da, the external IC to HepG2 hepatoma carcinoma cell50
Value is 1230.8 μ g/mL.
Each parameter of this comparative example is corresponding with embodiment 2, and the difference of the two is: embodiment 2 uses ionic liquid activation Asia
After selenic acid, the formamide solution with Fenugreek Polysaccharides reacts.
Table 1 is embodiment of the present invention 1-3 and comparative example is prepared selenizing Fenugreek Polysaccharides reaction temperature, response time and selenium and contained
The catalog of amount:
As can be seen from Table 1: Se content adds the reaction temperature in the formamide solution of Fenugreek Polysaccharides and reaction with Monohydrated selenium dioxide
Time is relevant.
Claims (9)
1. a synthetic method for selenizing Fenugreek Polysaccharides, comprises the following steps that:
(1) preparation of Fenugreek Polysaccharides formamide solution: by Fenugreek Polysaccharides at 60 DEG C ~ 80 DEG C, 300 ~ 500 rpm stirring 2 ~
5 h are dissolved in Methanamide, obtain the formamide solution of Fenugreek Polysaccharides；
(2) preparation of ionic liquid: concentrated sulphuric acid is added drop-wise to N-methyl pyrrole with the speed of 50 ~ 80/min at-10 DEG C ~ 0 DEG C
In pyrrolidone, 300 ~ 800 rpm stir reaction 10 ~ 18 h, wash 2 ~ 3 times by ethyl acetate after completion of the reaction, collect under room temperature
Lower floor's thick liquid, at 0.06 ~ 0.08 MPa, is vacuum dried 6 ~ 12 h at 50 ~ 70 DEG C and obtains ionic liquid；
(3) ionic liquid activation to Monohydrated selenium dioxide: at 60 ~ 80 DEG C and N2Under conditions of protection, Monohydrated selenium dioxide is joined above-mentioned (2)
Ionic liquid reacts the Monohydrated selenium dioxide after 1 ~ 3 h must activate；
(4) synthesis of selenizing Fenugreek Polysaccharides: by the formamide solution Dropping funnel of Fenugreek Polysaccharides that obtains in (1) with 30
The speed of ~ 60/min is added drop-wise in the Monohydrated selenium dioxide that (3) intermediate ion liquid activates, at 60 ~ 80 DEG C and N2Under conditions of protection,
React 2 ~ 4 h under 300 ~ 800 rpm, after question response, under conditions of isolation air, add washing with acetone 10 ~ 15 times, will
Flowing water dialysis 24 ~ 48 h, deionized water dialysis 12 ~ 24 in the bag filter of precipitate loading molecular cut off 8000 ~ 14000 Da
H, in complete back pkt. of dialysing thing under vacuum 0.06 ~ 0.08 MPa, 55 ~ 65 DEG C be evaporated to the 1/20 ~ 1/30 of original volume,
Adding dehydrated alcohol precipitation in concentrated solution, 4000 ~ 5000 rpm are centrifuged 10 ~ 15 min and collect lower sediment thing, 5 ~ 20
Pa, lyophilization 24 ~ 48 h at-60 ~-50 DEG C, obtain selenizing Fenugreek Polysaccharides.
2. the synthetic method of selenizing Fenugreek Polysaccharides as claimed in claim 1, it is characterised in that: in step (1), Fenugreek Polysaccharides
Weight average molecular weight be 2.303 × 106 Da, total sugar content is 87.2%.
The synthetic method of a kind of selenizing Fenugreek Polysaccharides the most as claimed in claim 1, it is characterised in that: in step (1), Semen Trigonellae
Polysaccharide is 2.5 ~ 4 mg/mL with the mass volume ratio of Methanamide.
The synthetic method of a kind of selenizing Fenugreek Polysaccharides the most as claimed in claim 1, it is characterised in that: in step (2), N-methyl
Ketopyrrolidine is 2:1 ~ 2.5:1 with the volume ratio of concentrated sulphuric acid.
The synthetic method of a kind of selenizing Fenugreek Polysaccharides the most as claimed in claim 1, it is characterised in that: in step (2), acetic acid second
The volume ratio of ester and ionic liquid is 0.5:1 ~ 1:1.
The synthetic method of a kind of selenizing Fenugreek Polysaccharides the most as claimed in claim 1, it is characterised in that: in step (3), Monohydrated selenium dioxide
It is 0.125 ~ 0.2 g/mL with the mass volume ratio of ionic liquid.
The synthetic method of a kind of selenizing Fenugreek Polysaccharides the most as claimed in claim 1, it is characterised in that: in step (4), Semen Trigonellae
The volume ratio of the formamide solution of polysaccharide and the Monohydrated selenium dioxide of activation is 10 ~ 25.
The synthetic method of a kind of selenizing Fenugreek Polysaccharides the most as claimed in claim 1, it is characterised in that: in step (4), acetone and
The volume ratio of reactant liquor is 2 ~ 3.
9. the selenizing Fenugreek Polysaccharides conduct prepared by the synthetic method of a kind of selenizing Fenugreek Polysaccharides described described in claim 1
The inhibitor of HepG2 hepatoma carcinoma cell is applied in cancer therapy drug.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610271490.8A CN105713104B (en) | 2016-04-28 | 2016-04-28 | A kind of synthesis and its application of selenizing Fenugreek Polysaccharides |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107573437A (en) * | 2017-10-12 | 2018-01-12 | 西北师范大学 | A kind of method of solid acid catalysis synthesis selenizing polysaccharide |
| CN107586347A (en) * | 2017-09-27 | 2018-01-16 | 西北师范大学 | A kind of method that round end wormwood artemisia polysaccharide derivates are synthesized using acidic ion liquid as catalyst |
| CN107629134A (en) * | 2017-09-18 | 2018-01-26 | 西北师范大学 | The method that selenizing artemisia desertorum polysaccharide is synthesized as solvent and catalyst using ionic liquid |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20050122289A (en) * | 2004-06-24 | 2005-12-29 | 정한춘 | Manufacturing process of chitosan-selenium complex |
| JP2008260823A (en) * | 2007-04-11 | 2008-10-30 | Univ Of Fukui | Polysaccharide modified with selenious acid and its manufacturing method |
| CN101421307A (en) * | 2006-03-27 | 2009-04-29 | 宝洁公司 | Methods for modifying biopolymers in ionic liquids |
| CN101624425A (en) * | 2009-08-05 | 2010-01-13 | 西北师范大学 | Method for preparing seleno-polysaccharide by organic method |
| CN101628946A (en) * | 2009-08-07 | 2010-01-20 | 西北师范大学 | Method for compounding polyhexose selenide by microwave |
| CN103193893A (en) * | 2013-04-18 | 2013-07-10 | 江南大学 | Preparation method of rice bran polysaccharide sulfate based on ionic liquid |
| CN103752340A (en) * | 2013-12-30 | 2014-04-30 | 浙江工业大学 | Applications of acidic functionalized ion liquid in esterification reaction |
-
2016
- 2016-04-28 CN CN201610271490.8A patent/CN105713104B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20050122289A (en) * | 2004-06-24 | 2005-12-29 | 정한춘 | Manufacturing process of chitosan-selenium complex |
| CN101421307A (en) * | 2006-03-27 | 2009-04-29 | 宝洁公司 | Methods for modifying biopolymers in ionic liquids |
| JP2008260823A (en) * | 2007-04-11 | 2008-10-30 | Univ Of Fukui | Polysaccharide modified with selenious acid and its manufacturing method |
| CN101624425A (en) * | 2009-08-05 | 2010-01-13 | 西北师范大学 | Method for preparing seleno-polysaccharide by organic method |
| CN101628946A (en) * | 2009-08-07 | 2010-01-20 | 西北师范大学 | Method for compounding polyhexose selenide by microwave |
| CN103193893A (en) * | 2013-04-18 | 2013-07-10 | 江南大学 | Preparation method of rice bran polysaccharide sulfate based on ionic liquid |
| CN103752340A (en) * | 2013-12-30 | 2014-04-30 | 浙江工业大学 | Applications of acidic functionalized ion liquid in esterification reaction |
Non-Patent Citations (1)
| Title |
|---|
| JUNLONG WANG等: ""Synthesis of selenium-containing polysaccharides and evaluation of antioxidant activity in vitro"", 《INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107629134A (en) * | 2017-09-18 | 2018-01-26 | 西北师范大学 | The method that selenizing artemisia desertorum polysaccharide is synthesized as solvent and catalyst using ionic liquid |
| CN107629134B (en) * | 2017-09-18 | 2019-12-31 | 西北师范大学 | Method for synthesizing selenylation artemisia desertorum polysaccharide by taking ionic liquid as solvent and catalyst |
| CN107586347A (en) * | 2017-09-27 | 2018-01-16 | 西北师范大学 | A kind of method that round end wormwood artemisia polysaccharide derivates are synthesized using acidic ion liquid as catalyst |
| CN107586347B (en) * | 2017-09-27 | 2019-12-31 | 西北师范大学 | Method for synthesizing artemisia sphaerocephala polysaccharide derivative by taking acidic ionic liquid as catalyst |
| CN107573437A (en) * | 2017-10-12 | 2018-01-12 | 西北师范大学 | A kind of method of solid acid catalysis synthesis selenizing polysaccharide |
| CN107573437B (en) * | 2017-10-12 | 2019-12-27 | 西北师范大学 | Method for synthesizing selenized polysaccharide by solid acid catalysis |
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| CN105713104B (en) | 2017-11-21 |
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