CN107721951B - The preparation method and application of 5 hydroxymethyl furfural trimer - Google Patents
The preparation method and application of 5 hydroxymethyl furfural trimer Download PDFInfo
- Publication number
- CN107721951B CN107721951B CN201711130605.2A CN201711130605A CN107721951B CN 107721951 B CN107721951 B CN 107721951B CN 201711130605 A CN201711130605 A CN 201711130605A CN 107721951 B CN107721951 B CN 107721951B
- Authority
- CN
- China
- Prior art keywords
- buffer
- trimer
- hydroxymethyl furfural
- cell
- control unit
- 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
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D307/00—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom
- C07D307/02—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom not condensed with other rings
- C07D307/34—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members
- C07D307/38—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members with substituted hydrocarbon radicals attached to ring carbon atoms
- C07D307/40—Radicals substituted by oxygen atoms
- C07D307/46—Doubly bound oxygen atoms, or two oxygen atoms singly bound to the same carbon atom
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2500/00—Screening for compounds of potential therapeutic value
- G01N2500/10—Screening for compounds of potential therapeutic value involving cells
Abstract
The present invention provides a kind of preparation method of 5 hydroxymethyl furfural trimer, the preparation method comprising steps of 1) into the organic solvent solution of 5 hydroxymethyl furfural be added catalytic amount acid, at 50~60 DEG C react 4~4.5h；Wherein, the organic solvent is one of chloroform, ethyl acetate, dimethylbenzene and toluene or a variety of；The acid is one of hydrochloric acid, sulfuric acid, acetic acid, trifluoroacetic acid and methanesulfonic acid or a variety of；2) reaction mixture is cooling and washes, and is concentrated to give crude product；3) gained crude product purifying, obtains 5 hydroxymethyl furfural trimer.The present invention also provides application of the 5 hydroxymethyl furfural trimer in reagent preparation and antiallergy screening compound.
Description
Technical field
The invention belongs to biomedicine technical fields, are related to a kind of preparation method of 5 hydroxymethyl furfural trimer and answer
With.
Background technique
Saccharide compound is the main source and the indispensable Major Foods of the mankind of body energy, with human lives
Closely related, e.g., cake, honey, coffee, wine etc. contain a large amount of saccharide compounds.Saccharide compound is also extensive in drug
Using if glucose injection, dextran make blood plasma preparation, starch makees tablet excipient, and Chinese medicine also contains a large amount of carbohydrate
Compound.Studies have shown that easily dehydration generates 5 hydroxymethyl furfural to monosaccharide under heating or acid condition, honey, wine etc. contain sugared object
A large amount of 5 hydroxymethyl furfural is detected in matter.5 hydroxymethyl furfural can be to groups such as eyes, the upper respiratory tract, skin and mucous membranes
Generation stimulation is knitted, there are the toxic side effects such as carcinogenic and teratogenesis, therefore pharmacopeia is using 5 hydroxymethyl furfural as containing the miscellaneous of sugared injection
Matter is defined.In addition, 5 hydroxymethyl furfural chemical property is active, it is easy to happen polymerization reaction, therefore, studies 5- methylol chaff
Safe handling of the polymerizate and toxic side effect of aldehyde to object containing confectionery and drug is ensured, ensures that human health has important meaning
Justice.
Summary of the invention
The purpose of the present invention is to provide a kind of preparation method and application of 5 hydroxymethyl furfural trimer, the preparation methods
Mild condition, process is easy to operate, and preparation cost is low；5 hydroxymethyl furfural trimer can promote rat basophilic leukemia
(RBL-2H3) cell release β-hexosaminidase and histamine.
The present invention is to be achieved through the following technical solutions:
A kind of preparation method of 5 hydroxymethyl furfural trimer, which is characterized in that comprising steps of
1) into the organic solvent solution of 5 hydroxymethyl furfural be added catalytic amount acid, at 50~60 DEG C react 4~
4.5h；Wherein, the organic solvent is one of chloroform, ethyl acetate, dimethylbenzene and toluene or a variety of；The acid is salt
One of acid, sulfuric acid, acetic acid, trifluoroacetic acid and methanesulfonic acid are a variety of；
2) reaction mixture is cooling and washes, and is concentrated to give crude product；
3) gained crude product purifying, obtains 5 hydroxymethyl furfural trimer；
Wherein, the structure of the 5 hydroxymethyl furfural trimer are as follows:
Preferably, in step 1), the concentration of the organic solvent solution of the 5 hydroxymethyl furfural is 0.1~0.6mol/
L。
Preferably, in step 1), sour dosage is the 0.1%~2% of organic solvent volume.
Preferably, in step 3), the method for crude product purifying are as follows: column chromatography for separation is carried out using silica gel, with acetic acid second
Ester: petroleum ether: triethylamine=1: for eluant, eluent eluted at 1.5: 0.16.
Application of the 5 hydroxymethyl furfural trimer in reagent preparation, the reagent can promote rat basophilic
Leukaemia cell discharges β-hexosaminidase and histamine.
Preferably, the reagent includes 5 hydroxymethyl furfural trimer and buffer, and the buffer includes 6.5~
7.3g/L NaCl, 0.31~0.37g/L KCl, 0.27~0.29g/L CaCl2, 0.13~0.15g/L MgSO4, 0.15~
0.18g/L KH2PO4, 2.2~2.5g/L HEPES, 0.9~1.1g/L glucose and 0.9~1.1g/LBSA, solvent is water；
Wherein, the concentration of 5 hydroxymethyl furfural trimer is 25~200 μM.
Preferably, the buffer includes 6.954g/L NaCl, 0.353g/L KCl, 0.282g/L CaCl2、
0.143g/L MgSO4、0.162g g/L KH2PO4, 2.383g/L HEPES, 0.991g/L glucose and 1.0g/L BSA.
A kind of screening technique of antiallergy compound, comprising steps of
1) culture experiment cell, experimental cell are randomly divided into cellular control unit and experimental group cell；Preparation control buffer
And test buffer；Wherein, it compares and contains 5 hydroxymethyl furfural trimer in buffer；Contain to be testedization in test buffer
Object is closed, remaining component is identical as control buffer；
2) control buffer is added into cellular control unit, test buffer is added into experimental group cell；Two kinds of cells
It cultivates at identical conditions；
3) after cell culture, the beta-amino hexose in the buffer of cellular control unit and experimental group cell is tested respectively
Glycosides enzyme r e lease rate；
If the β-hexosaminidase release rate in the buffer of cellular control unit is greater than experimental group cell, to be testedization
Closing object has antiallergic activity；
If the β-hexosaminidase release rate in the buffer of cellular control unit is not more than experimental group cell, to be tested
Compound does not have antiallergic activity.
Preferably, in step 3), after cell culture, the buffering of cellular control unit and experimental group cell is tested respectively
Histamine release amount in liquid；
If the histamine release amount in the buffer of cellular control unit is greater than experimental group cell, compound to be tested has anti-
Activity；
If the histamine release amount in the buffer of cellular control unit is not more than experimental group cell, compound to be tested does not have
There is antiallergic activity.
Preferably, compare the component of buffer are as follows: 6.5~7.3g/L NaCl, 0.31~0.37g/L KCl, 0.27~
0.29g/L CaCl2, 0.13~0.15g/L MgSO4, 0.15~0.18g/L KH2PO4, 2.2~2.5g/L HEPES, 0.9~
1.1g/L glucose and 0.9~1.1g/L BSA, solvent are water；Wherein, the concentration of 5 hydroxymethyl furfural trimer be 25~
200μM。
Compared with prior art, the invention has the following beneficial technical effects:
The preparation method of 5 hydroxymethyl furfural trimer provided by the invention, raw material sources are easy to get, and reaction condition is mild,
Reaction process is easy to operate, and agents useful for same is cheap and easily-available.
The application of 5 hydroxymethyl furfural trimer provided by the invention, prepared reagent can promote RBL-2H3 cell
β-hexosaminidase and histamine are discharged, can be used for the research of allergic reaction containing sugar substance.
The screening technique of antiallergy compound provided by the invention, it can be used to quick primary dcreening operation antiallergy substances, find
Antiallergy lead compound.
Detailed description of the invention
Fig. 1 is the synthetic route of 5 hydroxymethyl furfural trimer provided by the invention.
Fig. 2 is after being acted on respectively with 25 μM, 50 μM, 100 μM, 200 μM of 5 hydroxymethyl furfural trimer, and RBL-2H3 is thin
The β-hexosaminidase releasing result of born of the same parents.
Fig. 3 is after being acted on respectively with 25 μM, 50 μM, 100 μM, 200 μM of 5 hydroxymethyl furfural trimer, and RBL-2H3 is thin
The histamine release result of born of the same parents.
Specific embodiment
Below with reference to specific embodiment, the present invention is described in further detail, it is described be explanation of the invention and
It is not to limit.
5 hydroxymethyl furfural trimer provided by the invention can promote RBL-2H3 cell release β-hexosaminidase and
Histamine, chemical structural formula are as follows:
Below with reference to synthetic route shown in Fig. 1 and specific synthesis embodiment the present invention will be described in detail 5- hydroxyl first
The preparation method and application of base furfural trimer.
Embodiment 1:
As shown in Figure 1, a kind of 5 hydroxymethyl furfural trimer the preparation method is as follows:
1) 5 hydroxymethyl furfural (2.52g, 20.00mmol) is placed in 100mL round-bottomed flask, and it is molten that 50mL chloroform is added
Solution, is added dropwise 0.05mL concentrated hydrochloric acid, and 4~4.5h is reacted in 50 DEG C of -60 DEG C of heating.
2) it is cooled to room temperature, washing removes after 5 hydroxymethyl furfural (thin-layer chromatography monitoring, about 10 × 50mL), successively with full
With saline solution (2 × 50mL) washing, the dry organic phase of anhydrous sodium sulfate, removing solvent is concentrated under reduced pressure and obtains crude product.
3) crude product carries out column chromatography for separation using silica gel, and with ethyl acetate: petroleum ether: triethylamine=1: be at 1.5: 0.16
Eluant, eluent is eluted, and R is collectedfSample eluent at value about 0.16, is concentrated to give 5 hydroxymethyl furfural trimer 63mg, produces
Rate 2.63%.
Gained compound is colourless oil liquid, molecular formula C18H16O8。ESI-MS(m/z):383.10[M+Na]+,1H
NMR(400MHz,CDCl3): δ 9.61 (s, 2H), 7.22 (d, J=3.5Hz, 2H), 6.56 (d, J=3.5Hz, 2H), 6.47 (d,
J=3.1Hz, 1H), 6.29 (d, J=3.2Hz, 1H), 5.79 (s, 1H), 4.68 (s, 4H), 4.60 (s, 2H)13C NMR
(101MHz,CDCl3):δ177.74,157.36,155.07,152.73,149.12,125.53,111.85,110.53,
108.30,95.81,59.45,57.30.
Embodiment 2:
The configuration of TM buffer:
Weigh 6.5~7.3g NaCl, 0.31~0.37g KCl, 0.27~0.29g CaCl2, 0.13~0.15gMgSO4、
0.15~0.18g KH2PO4, 2.2~2.5g HEPES, 0.9~1.1g glucose and 0.9~1.1gBSA, be added tri-distilled water
800mL, stirring and dissolving are added tri-distilled water to final volume 1L, are removed in super-clean bench with sterilized 0.22 μm of filtering with microporous membrane
Bacterium saves at 4 DEG C.
As a kind of specific situation, in the present embodiment, the concentration of each component in TM buffer are as follows: 6.954g/L
NaCl、0.353g/L KCl、0.282g/L CaCl2、0.143g/L MgSO4、0.162g g/L KH2PO4、2.383g/L
HEPES, 0.991g/L glucose and 1.0g/L BSA.
According to method as above, the TM buffer solution containing 5 hydroxymethyl furfural trimer is prepared, wherein 5 hydroxymethyl furfural
Trimer is dissolved in TM buffer；In the prepared TM buffer solution containing 5 hydroxymethyl furfural trimer, 5- methylol chaff
The concentration of aldehyde trimer is respectively 25 μM, 50 μM, 100 μM, 200 μM.
The configuration of terminate liquid:
1.055g Na2CO3, it is dissolved in 100mL tri-distilled water, obtains the Na of 0.1mol/L2CO3Solution is protected from light guarantor at 4 DEG C
It deposits.Weigh 0.84g NaHCO3, it is dissolved in 40mL tri-distilled water, obtains the NaHCO of 0.1mol/L3Solution is kept in dark place at 4 DEG C.
Before use, by 0.1mol/L Na2CO3: 0.1mol/L NaHCO3The ratio of=9:1 (v/v) mixes, and obtains 0.1mol/L
Na2CO3/NaHCO3Terminate liquid.
β-hexosaminidase release rate assay experiment: to logarithmic phase well-grown RBL-2H3 cell (adherent life of cell
Long, be in shuttle shape, arrangement is close) it is long to culture bottle floor space 80% or so when, inhale and abandon culture medium, PBS is added, clean three times,
The 0.25% trypsin solution digestion containing EDTA is added, serum-containing media terminates, and collects cell, cell count, with containing blood
Cell is diluted to final concentration of 5 × 10 by clear culture medium5The cell suspension of a/mL, every 100 μ L of hole, which is inoculated in 96 orifice plates, (to be made every
The final cell concentration in hole is 5 × 104A cell).96 orifice plates are placed in the cell incubator of 37 DEG C of saturated humidities, were cultivated
Night keeps cell adherent.
It next day, inhales and abandons culture medium, 100 μ L TM buffer solutions are cleaned twice, are separately added into not according to different cells
Same reagent:
Administration group is separately added into containing final concentration of 25 μM, 50 μM, 100 μM, 200 μM of 5 hydroxymethyl furfural trimer
100 μ L TM buffer solutions；
The 100 μ L TM buffer solutions of positive group (C48/80) the final concentration of 30 μ g/mL C48/80 of addition；
100 μ L TM buffer solutions are added in blank group (control)；
After reagent is added, after cultivating 30min at 37 DEG C, 10min is terminated on ice, and 1000rpm is centrifuged 10min at 4 DEG C, obtains
The cell incubation liquid supernatant of corresponding cell grouping takes the cell incubation liquid supernatant of the corresponding cell grouping of 50 μ L to 96 hole of blank
Plate；Exhaust blank group cell conditioned medium, then use 0.1%TritonX-100, blow and beat 10 times, crack blank group cell 5min, on ice end
Only 10min, 1000rpm is centrifuged 10min at 4 DEG C, obtains cracking group supernatant, takes 50 μ L cracking group supernatants to 96 orifice plate of blank.
Administration group supernatant (administration group cell incubation liquid supernatant), positive group supernatant (positive group cell incubation liquid supernatant), sky
The 50 μ L of beta-amino hexose of 1mmol/L is added in white group of supernatant (blank group cell incubation liquid supernatant) and the every hole of cracking group supernatant, sets
In being incubated for 90min in 37 DEG C of incubators, after the completion of incubation, 150 μ L Na are added in every hole2CO3/NaHCO3(0.1mol/L Na2CO3∶
0.1mol/L NaHCO3=9: 1 (v/v), pH=11) terminate liquid termination reaction.96 orifice plates are placed on room temperature shaker rock it is mixed
Even 2min measures absorbance value (OD) under microplate reader 405nm wavelength.The calculation formula of β-hexosaminidase release rate is such as
Under:
Wherein, ODSupernatantFor ODBlank group supernatant、ODAdministration group supernatantOr ODPositive group supernatant。
Experimental results show is in Fig. 2, the results showed that, 5 hydroxymethyl furfural trimer can promote RBL-2H3 cell
β-hexosaminidase release.
Embodiment 3:
Histamine release experiments: to the well-grown RBL-2H3 cell of logarithmic phase, (cell adherent growth, is in shuttle shape, and arrangement is tight
It is close) it is long to culture bottle floor space 80% or so when, inhale and abandon culture medium, PBS is added, clean three times, be added containing EDTA's
The digestion of 0.25% trypsin solution, serum-containing media terminate, and collect cell, cell count will be thin with serum-containing media
Born of the same parents are diluted to final concentration of 5 × 105The cell suspension of a/mL, every 100 μ L of hole are inoculated in the 96 orifice plates (cell for keeping every hole final
Amount is 5 × 104A cell).96 orifice plates are placed in the cell incubator of 37 DEG C of saturated humidities, overnight incubation, keep cell adherent.
Next day inhales and abandons culture medium, and 100 μ L TM buffer solutions clean twice, and different examinations is separately added into according to different cells
Agent:
Administration group is separately added into containing final concentration of 25 μM, 50 μM, 100 μM, 200 μM of 5 hydroxymethyl furfural trimer
100 μ L TM buffer solutions；
The 100 μ L TM buffer solutions of positive group (C48/80) the final concentration of 30 μ g/mL C48/80 of addition；
Negative group (control) 100 μ L TM buffer solutions of addition；
At 37 DEG C cultivate 30min after, terminate 10min on ice, at 4 DEG C 1000rpm be centrifuged 10min, respectively obtain administration group,
Feminine gender group and positive group cell conditioned medium.Take 50 μ L supernatants into 1.5mL EP pipe respectively, the deuterated histamine of every pipe addition be (5ng/mL's
Acetonitrile solution) 100 μ L are as internal standard.After mixed solution vortex oscillation, 12000g is centrifuged 20min at 4 DEG C, takes supernatant, UHPLC-
ESI-MS/MS measurement.Basic chromatographic condition are as follows: chromatographic column is HILIC column (Venusil HILIC, 2.1 × 150mm, 3 μm).
Experimental results show is in Fig. 3, the results showed that, 5 hydroxymethyl furfural trimer can promote RBL-2H3 cell to release
Histamine is put, and burst size and activity are positively correlated.
Embodiment 4
A kind of screening technique of antiallergy compound, comprising steps of
1) culture experiment cell, experimental cell are randomly divided into cellular control unit and experimental group cell；
It is long to culture bottle to the well-grown RBL-2H3 cell of logarithmic phase (cell adherent growth, is in shuttle shape, and arrangement is close)
Floor space 80% or so when, inhale abandon culture medium, be added PBS, clean three times, be added 0.25% trypsin solution containing EDTA
Digestion, serum-containing media terminate, and collect cell, cell count, cell is diluted to final concentration of 5 with serum-containing media ×
105The cell suspension of a/mL, every 100 μ L of hole, which is inoculated in 96 orifice plates, (makes the cell concentration 5 × 10 that every hole is final4A cell).It will
96 orifice plates are placed in the cell incubator of 37 DEG C of saturated humidities, overnight incubation, keep cell adherent.
Next day inhales and abandons culture medium, gained cell is divided into cellular control unit and experimental group cell.
2) preparation control buffer and test buffer；Wherein, it compares and contains 5 hydroxymethyl furfural trimer in buffer；
Contain compound to be tested in test buffer, remaining component is identical as control buffer.
Weigh 6.5~7.3g NaCl, 0.31~0.37g KCl, 0.27~0.29g CaCl2, 0.13~0.15g
MgSO4, 0.15~0.18g KH2PO4, 2.2~2.5g HEPES, 0.9~1.1g glucose and 0.9~1.1g BSA, be added
Tri-distilled water 800mL, stirring and dissolving add tri-distilled water to final volume 1L, with sterilized 0.22 μm of miillpore filter in super-clean bench
Filtration sterilization saves at 4 DEG C, obtains buffer.
5 hydroxymethyl furfural trimer is dissolved in buffer, control buffer is obtained；
5 hydroxymethyl furfural trimer and compound to be tested are dissolved in buffer, test buffer is obtained；
Wherein, control buffer is identical with the 5 hydroxymethyl furfural trimer concentration in test buffer, and concentration range is
25~200 μM.
3) control buffer is added into cellular control unit, test buffer is added into experimental group cell；It is trained at 37 DEG C
After supporting 30min, 10min is terminated on ice.
4) β-hexosaminidase release rate is measured according to 2 the method for embodiment.
If the β-hexosaminidase release rate in the buffer of cellular control unit is greater than experimental group cell, to be testedization
Closing object has antiallergic activity；
If the β-hexosaminidase release rate in the buffer of cellular control unit is not more than experimental group cell, to be tested
Compound does not have antiallergic activity.
Embodiment 5 is referring to 4 the method for embodiment, wherein in step 4), according to 3 the method measurement group of embodiment
Amine burst size.
If the histamine release amount in the buffer of cellular control unit is greater than experimental group cell, compound to be tested has anti-
Activity；
If the histamine release amount in the buffer of cellular control unit is not more than experimental group cell, compound to be tested does not have
There is antiallergic activity.
To sum up, the preparation method of 5 hydroxymethyl furfural trimer provided by the invention is easy to get with raw material sources, reacts item
Part is mild, and reaction process is easy to operate, the cheap and easily-available advantage of agents useful for same.5 hydroxymethyl furfural trimer can promote RBL-
2H3 cell discharges β-hexosaminidase and histamine, and with dosage correlation.
Claims (6)
1. a kind of application of 5 hydroxymethyl furfural trimer in reagent preparation, which is characterized in that the reagent can promote greatly
Mouse basophilic leukemia cell discharges β-hexosaminidase and histamine；Wherein, the structure of 5 hydroxymethyl furfural trimer is as follows:
2. application of the 5 hydroxymethyl furfural trimer as described in claim 1 in reagent preparation, which is characterized in that the examination
Agent includes 5 hydroxymethyl furfural trimer and buffer, and the buffer includes 6.5~7.3g/L NaCl, 0.31~0.37g/L
KCl, 0.27~0.29g/L CaCl2, 0.13~0.15g/L MgSO4, 0.15~0.18g/L KH2PO4, 2.2~2.5g/L
HEPES, 0.9~1.1g/L glucose and 0.9~1.1g/L BSA, solvent is water；Wherein, 5 hydroxymethyl furfural trimer
Concentration is 25~200 μM.
3. application of the 5 hydroxymethyl furfural trimer as claimed in claim 2 in reagent preparation, which is characterized in that described slow
Fliud flushing includes 6.954g/L NaCl, 0.353g/L KCl, 0.282g/L CaCl2、0.143g/L MgSO4、0.162g g/L
KH2PO4, 2.383g/L HEPES, 0.991g/L glucose and 1.0g/L BSA.
4. a kind of screening technique of Claritin, which is characterized in that comprising steps of
1) RBL-2H3 cell is cultivated, cellular control unit and experimental group cell are randomly divided into；Preparation control buffer and experiment buffering
Liquid；Wherein, it compares and contains 5 hydroxymethyl furfural trimer in buffer；Contain 5 hydroxymethyl furfural trimer in test buffer
With drug to be tested, remaining component with control buffer it is identical；The structure of 5 hydroxymethyl furfural trimer is as follows:
2) control buffer is added into cellular control unit, test buffer is added into experimental group cell；Two kinds of cells are in phase
It is cultivated under conditions of；
3) after cell culture, the β-hexosaminidase in the buffer of cellular control unit and experimental group cell is tested respectively
Release rate；
If the β-hexosaminidase release rate in the buffer of cellular control unit is greater than experimental group cell, drug tool to be tested
There is antiallergic activity；
If the β-hexosaminidase release rate in the buffer of cellular control unit is not more than experimental group cell, drug to be tested
Without antiallergic activity.
5. the screening technique of Claritin as claimed in claim 4, which is characterized in that in step 3), cell culture knot
Shu Hou tests the histamine release amount in the buffer of cellular control unit and experimental group cell respectively；
If the histamine release amount in the buffer of cellular control unit is greater than experimental group cell, drug to be tested is living with antiallergy
Property；
If the histamine release amount in the buffer of cellular control unit is not more than experimental group cell, drug to be tested does not have anti-mistake
Quick activity.
6. the screening technique of Claritin as described in claim 4 or 5, which is characterized in that compare the component of buffer are as follows:
6.5~7.3g/L NaCl, 0.31~0.37g/L KCl, 0.27~0.29g/L CaCl2, 0.13~0.15g/L MgSO4、
0.15~0.18g/L KH2PO4, 2.2~2.5g/L HEPES, 0.9~1.1g/L glucose and 0.9~1.1g/L BSA, it is molten
Agent is water；Wherein, the concentration of 5 hydroxymethyl furfural trimer is 25~200 μM.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711130605.2A CN107721951B (en) | 2017-11-15 | 2017-11-15 | The preparation method and application of 5 hydroxymethyl furfural trimer |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711130605.2A CN107721951B (en) | 2017-11-15 | 2017-11-15 | The preparation method and application of 5 hydroxymethyl furfural trimer |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107721951A CN107721951A (en) | 2018-02-23 |
| CN107721951B true CN107721951B (en) | 2019-07-23 |
Family
ID=61216098
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711130605.2A Active CN107721951B (en) | 2017-11-15 | 2017-11-15 | The preparation method and application of 5 hydroxymethyl furfural trimer |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107721951B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110806374B (en) * | 2019-09-29 | 2022-05-20 | 杭州联科生物技术股份有限公司 | Binding Buffer for detecting Annexin V apoptosis and preparation method thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101522655A (en) * | 2006-10-06 | 2009-09-02 | 高砂香料工业株式会社 | Taste-improving agent and food or drink containing the same |
| CN101544624A (en) * | 2009-05-11 | 2009-09-30 | 吉林大学 | 5-methylol-furfural-5-furfural-methanol, preparation method thereof and medical application thereof |
| CN103257237A (en) * | 2013-05-09 | 2013-08-21 | 中国农业大学 | In-vitro detection method of allergen in food |
-
2017
- 2017-11-15 CN CN201711130605.2A patent/CN107721951B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101522655A (en) * | 2006-10-06 | 2009-09-02 | 高砂香料工业株式会社 | Taste-improving agent and food or drink containing the same |
| CN101544624A (en) * | 2009-05-11 | 2009-09-30 | 吉林大学 | 5-methylol-furfural-5-furfural-methanol, preparation method thereof and medical application thereof |
| CN103257237A (en) * | 2013-05-09 | 2013-08-21 | 中国农业大学 | In-vitro detection method of allergen in food |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107721951A (en) | 2018-02-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101260097B (en) | Technique for preparing potassium sodium dehydroandroandrographolide succinic by using potassium dehydroandrographolide succinate | |
| CN106588928B (en) | Heptacyclic compound, synthesis, activity rating and application | |
| CN101357146B (en) | Preparation method of chimonanthus nitens valid target, production method and use of formulation thereof | |
| CN105566317B (en) | A kind of compound and preparation method thereof | |
| CN101260098B (en) | Technique for preparing potassium sodium dehydroandroandrographolide succinate | |
| CN102302685B (en) | Common lophatherum herb extract and preparation method and application thereof | |
| CN107721951B (en) | The preparation method and application of 5 hydroxymethyl furfural trimer | |
| CN101590128B (en) | Lipid regulating and antioxidant effect-based method for evaluating quality of Jiangzhining | |
| CN106977467A (en) | A kind of compound and preparation method thereof and purposes | |
| CN101966172B (en) | New purpose of caffeic acid and derivatives thereof | |
| CN109061031A (en) | Alexandrium mimutum Halim metabolite-gonyatoxin isolation and purification method | |
| CN106966997A (en) | A kind of compound and preparation method thereof and purposes | |
| CN101805385B (en) | Neoflavonoid compounds separated and purified from Tridax procumbens and preparation method thereof | |
| CN101966194A (en) | New application of scutellarin and derivatives thereof | |
| CN103014090B (en) | Method for extracting bis-benzene oxepin-11(6H) keto-1, 10-dihydroxy, 3-methyl-7, 8-dimethoxy from Moller bacteria | |
| CN102144966B (en) | Preparation method of clindamycin phosphate injection | |
| CN101293889B (en) | Water-soluble arteannuin derivative and preparation method thereof | |
| CN102250176B (en) | Antitumor antibiotic ancomycin and its derivative | |
| CN110117630A (en) | A kind of method that preparation phenethyl caffeate glucosides was applied and was used in the glycosylation of dextransucrase | |
| CN102718807B (en) | Novel chiral double-site chitin synthesis inhibitor insecticide | |
| CN101670074A (en) | Preparation method for antivirus compound | |
| CN101348514B (en) | Use of starfish saponin compound extracted from Culcita novaeguineae | |
| CN100497363C (en) | Lactose with first anti-adhesion activity and its fluorescent mark | |
| CN104292351A (en) | Phellinus ribis polysaccharide, as well as preparation method and applications thereof | |
| CN105330563B (en) | For the reference compound in the control of Kukoamine B quality |
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 |