CN107085004A - A kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on Ni Gd C nanos probe - Google Patents
A kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on Ni Gd C nanos probe Download PDFInfo
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- CN107085004A CN107085004A CN201710228321.0A CN201710228321A CN107085004A CN 107085004 A CN107085004 A CN 107085004A CN 201710228321 A CN201710228321 A CN 201710228321A CN 107085004 A CN107085004 A CN 107085004A
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N24/00—Investigating or analyzing materials by the use of nuclear magnetic resonance, electron paramagnetic resonance or other spin effects
- G01N24/08—Investigating or analyzing materials by the use of nuclear magnetic resonance, electron paramagnetic resonance or other spin effects by using nuclear magnetic resonance
- G01N24/088—Assessment or manipulation of a chemical or biochemical reaction, e.g. verification whether a chemical reaction occurred or whether a ligand binds to a receptor in drug screening or assessing reaction kinetics
Abstract
A kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on Ni Gd C nanos probe, belongs to tcm development applied technical field.The present invention dependent on set up based on prepares Ni Gd C nanos probe be used for screen suppression anaerobic bacteria Chinese medicine preparation method.The characteristics of paramagnetic Ni Gd C nanos probes prepared can be specifically bound to object bacteria is coated with using antibody, the influence using Ni Gd C paramagnetic properties to the NMR relaxation time detects whether contain object bacteria in sample.Paramagnetic Ni Gd C nano probes, NMR relaxation time showing outlet sexual intercourse within the specific limits, i.e. Ni nanoparticle Gd C contents are big, the nuclear magnetic resonance spin Lattice Relaxation Time and transverse relaxation time value of sample are smaller, detection object bacteria can be quantified in certain limit, so that the fungistatic effect of indirect assessment Chinese medicine preparation.This method can be used for the recipe determination for having the Chinese medicine of inhibitory action to anaerobic bacteria, accelerate the development rate of tcm product.
Description
Technical field
The present invention relates to a kind of screening technique of Chinese medicine preparation.
Background technology
Antibiotic simultaneously, will also result in human body infringement, influence liver, renal function, gastrointestinal reaction etc. in sterilization.Abuse is anti-
Raw element may cause the generation of some bacterial resistance phenomenons, and the treatment to infection can become very difficult.At present, almost without one
Plant antibacterials and resistance phenomenon is not present.Chinese medicine and formula of the exploitation with antibacterial action have positive effect.
Many Chinese medicines and magistral medicines have antibacterial and anti-inflammation functions, and some medium-height grass the effective elements of the medicines and molecular structure etc. are also
Through studying clear whole or in part.The main component berberine of such as coptis and golden cypress stop dysentery（Berberine）, radix scutellariae antibacterial
Main component baicalein etc..A host of facts prove, the medicine that ancient Chinese working people is accumulated by long-term practice
Legacy is extremely abundant, extremely valuable.We should value the great treasure-house of this motherland's medicine and pharmacology, make great efforts to excavate, be subject to
Improve, can therefrom develop the Chinese medicine with antibacterial and anti-inflammation functions, can replace or part substitution antibiotic.
Anaerobic bacteria be a class under anaerobic than the bacterium grown in aerobic environment, and can not be with the presence of air
In the case of the bacterium that grows.This kind of bacterium lacks complete metabolism enzyme system, and its energetic supersession is carried out in the way of anaerobic fermentation.
It can cause the infection of human body different parts, including appendicitis, cholecystitis, tympanitis, mouth infection, endocarditis, osteomyelitis,
A variety of diseases such as peritonitis.It is usually in sterilizing in Chinese medicine new formulation procedures of traditional exploitation for certain anaerobic bacteria
Traditional Chinese medicinal components are added after inoculation of medium anaerobic bacteria, are cultivated in special anaerobic culture box.Sampled again after a period of time into
Row line culture, calculates clump count, so that it is determined that whether certain Chinese medicine compositing formula has bacteriostasis.Line culture is also required to detesting
Cultivated in oxygen incubator, and cultivation cycle at least takes several days, therefore whole cycle is long, efficiency is low.This method is asked for above-mentioned
Inscribe a kind of rapid screening method of Chinese medicine preparation of invention.
The general principle of this method：Monoclonal antibody or antigen molecule are by Covalent bonding together, and this combination will not change
Monoclonal antibody, the immunological characteristic of antigen and bioactivity, specific monoclonal antibody only can be with specific antigen knots
Close.And Ni-Gd-C materials have ferromagnetism, its outer layer has multiple unpaired electrons, with high saturation and magnetic intensity, in particle diameter
It is small to a certain extent when, it may appear that superparamagnetic characteristic, Ni-Gd-C nano materials can to the nuclear magnetic resonance spins of surrounding water molecules-
The influence of Lattice Relaxation Time is very big, the effect with amplification nuclear magnetic resonance relaxation signals.Very micro Ni-Gd-C nanometers
The NMR relaxation time that material will result in water declines to a great extent.Therefore can be by building Ni-Gd-C nano-probes, from magnetic
The angle of resonance detects whether to have object bacteria, so that the bacteriostasis of indirect assessment traditional Chinese medicine ingredients.
Key step：
1）Prepared by Ni-Gd-C nano-probes.Commercially paramagnetic Ni-Gd-C nano materials, can also pass through its other party
Method prepares nano level Ni-Gd-C.Using silane coupler, its formula is:Y(CH2)nSiX3.Herein, n is 0-3；X is can water
The group of solution；Y is organo-functional group.X is typically chloro, methoxyl group, ethyoxyl, acetoxyl group etc., when these groups are hydrolyzed
Generate silanol (Si (OH)3), and combined with inorganic substances, form siloxanes.Y is vinyl, amino, epoxy radicals, metering system
Acyloxy, sulfydryl.These reactive groups can react and combine with organic substance.Therefore, can be in nothing by using silane coupler
" molecular bridge " is erected between machine material and the interface of organic substance, the material of two kinds of property great disparities is linked together and improves compound
The effect of the performance and increase adhesive strength of material.The amination or carboxylic of surface modification can be obtained by silane coupler reaction
The Ni-Gd-C nano materials of base.Further by coupled antibody, surface-functionalized, formation specific immunity probe can be achieved,
The unnecessary avtive spot of closing, can separate excessive antibody after centrifugation, you can to obtain the Ni-Gd-C of resistance modification
Nano-probe.
2）The sample cell made using polystyrene material carries out Chinese medicine screening.Polystyrene material and ELISA Plate class
Seemingly, detecting probe surface antibody can be combined on the surface of polystyrene material due to suction-operated, still, and the antibody of detecting probe surface is inhaled
The speed of polystyrene material surface is attached to compared with the speed of detecting probe surface antibody binding object bacteria, the former is slow-footed more,
Both are competitive relations.Therefore, the culture medium of the anaerobic bacteria of sterilizing, vaccination target anaerobic bacteria are added in polystyrene sample pipe
The Chinese medicine extract for needing to screen is added afterwards, after closing anaerobic culture for a period of time, adds step 1）Prepared Ni-Gd-C receives
Rice probe, is sufficiently mixed concussion reaction for a period of time, if there is target bacteria growing in sample cell, Ni-Gd-C nano-probes surface
Antibody can be combined first with object bacteria so that Ni-Gd-C nano-probes are stayed in a liquid.If there is no target in sample cell
Bacterium, then the antibody on Ni-Gd-C nano-probes surface can gradually adsorb in polystyrene sample pipe surface so that not having in liquid
Ni-Gd-C nano-probes.Preceding to have addressed, the NMR relaxation time that micro Ni-Gd-C nano materials will result in water is big
Width declines.Using the sterile medium liquid without vaccination target bacterium as blank, the relaxation time of the suspension measured, which is compared, to be had
Significantly reduce, then illustrate to contain probe in liquid, so as to there is object bacteria in indirect proof sample, the amount of probe and relaxation time
Drop-out value is proportional, and the amount of object bacteria can be gone out with indirect quantification by the quantitative probe of mark-on.With the presence of object bacteria, then illustrate Chinese medicine
Bacteriostasis is not obvious, it is on the contrary then explanation have bacteriostasis.
The major advantage of this method is exactly quick, sensitivity height.Object bacteria can be realized and continuously existed to a certain extent
Line is monitored.Time of even several days 2-3 days relative to anaerobic bacteria culture, magnetic resonance detection only needs a few minutes.The method is not required to
Anaerobic culture box is wanted, can cultivate the sample seal of tube after need to being only inoculated with anaerobic, will be quantitative with syringe during detection
Probe injection can carry out follow-up magnetic resonance detection.Therefore, the screening of extensive Chinese medicine and formula can be made of this method.
The content of the invention
A kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on Ni-Gd-C nano-probes.This method, which is that one kind is objective, to be had
The method of object bacteria, can do the quick screening of extensive Chinese medicine and formula, so as to a certain extent in the detection sample of effect
Reduction is directed to certain anaerobic bacteria effective tcm development time significantly.
A kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on Ni-Gd-C nano-probes.Using NMR to suitable
The response sensibility of magnetic material, proposes NMR relaxation Parameters variation and Ni-Gd-C nano-particle paramagnetic immunological probe contents
Correlation metric.This method is used to the detection of object bacteria in sample assess dependent on paramagnetic nano Ni-Gd-C nano-probes
The fungistatic effect of Chinese medicine preparation., can be to sample using the paramagnetic Ni-Gd-C nano-probes for being coupled monoclonal antibody specific
In specific anaerobic bacteria detected.Because NMR is very sensitive to Ni-Gd-C nano-particles, exist micro suitable
Magnetic Ni-Gd-C nano-particles, then the spin-lattice relaxation time of water and/spin-spin relaxation will be remarkably decreased.Pass through sky
White control and mark-on quantitatively detect the immunological probe content in sample, the anaerobic bacteria bacterium gone out so as to indirect quantification in sample
Fall number.The anaerobism bacterial content detected and probe content linear correlation, degree of fitting are preferable.Finally with Ni-Gd-C nanometers of spies of paramagnetic
Corresponding relation between pin and anaerobic bacteria is tie, determines the anaerobic bacteria clump count in sample.So as to determine the antibacterial of Chinese medicine indirectly
Whether effect is effective.Time of even several days 2-3 days relative to anaerobic bacteria culture, magnetic resonance detection only needs a few minutes.Therefore,
The screening of extensive Chinese medicine and formula can be made of this method.
The present invention is achieved in that step is as follows：
1）Target bacteria antibody is coated with the preparation of paramagnetic nano Ni-Gd-C nano-probes.
2）The culture medium of the anaerobic bacteria of sterilizing, vaccination target anaerobic bacteria are added in polystyrene sample pipe.Adding needs
The Chinese medicine extract of screening, after culture a period of time, after culture a period of time, adds step 1）Prepared Ni-Gd-C nanometers
Probe, is sufficiently mixed concussion reaction for a period of time, if there is object bacteria in sample cell, first can be with Ni-Gd-C nano-probe tables
The antibody binding in face, so that Ni-Gd-C nano-probes are stayed in a liquid.If not having object bacteria in sample cell, Ni-
The antibody on Gd-C nano-probes surface can gradually be adsorbed and separated in polystyrene sample pipe surface with liquid.Now, to sample
Pipe carries out the measure of NMR relaxation time.Using the sterile medium liquid without vaccination target bacterium as blank, measure
The relaxation time of suspension, then explanation contained probe, so as to have object bacteria, probe in indirect proof sample compared to significantly reducing
Amount and the decline in relaxation time it is proportional, by mark-on quantitative probe, indirect quantification goes out the amount of object bacteria.
The Ni-Gd-C nano-probes are the nanoscale Ni-Gd-C materials with paramagnetic properties, and nanometer particle size is less than 300
Nanometer.
The change in final relaxation time of the detection evaluation method based on nuclear magnetic resonance technique of described object bacteria, during relaxation
Between include spin-lattice relaxation time and spin spin relaxation time.
Beneficial effects of the present invention：The present invention, which provides a kind of objectively quick screening, the Chinese medicine of inhibitory action to anaerobic bacteria
The method of formula, it is characterized in that using the corresponding relation between paramagnetic Ni-Gd-C nano-probes and anaerobic bacteria as tie, determining in sample
Anaerobic bacteria clump count.So as to determine whether the bacteriostasis of Chinese medicine is effective indirectly.It is even several relative to anaerobic bacteria culture 2-3 days
It time, magnetic resonance detection only needs a few minutes.Therefore, the screening of extensive Chinese medicine and formula can be made of this method.
Embodiment
Example 1
Screening is to Propiobacterium（Anaerobic bacteria）There is the Chinese medicine preparation of suppression or therapeutic action.
1）It is prepared by Ni nanoparticle-Gd-C immunological probes：
Ni-Gd-C nano-particles can be commercially（Such as the Chao Wei nano materials company in Shanghai）, can also bibliography
With either physically or chemically synthesizing.
Silicon dioxide coated Ni nanoparticle-Gd-C：47.5g sodium metasilicate is taken, is dissolved in deionized water in beaker, is adjusted with hydrochloric acid
Section pH value is 12-13.5.0g Ni nanoparticles-Gd-C is taken to be added in this beaker, mechanical agitation (using glass bar) 5min.By mixed liquor
Ultrasonic 30min, in good time stirring.85 C are warming up to, salt acid for adjusting pH value 6-7, generation precipitation is added dropwise.Spent in Magneto separate
Ion water washing is precipitated, and is washed 3-4 times.Then, precipitation is scattered in 250mL methanol.Above procedure is in triplicate, it is ensured that silicon
It is attached on Ni-Gd-C.
Amino containing silane Ni-Gd-C nano materials：Obtained silicon dioxide coated Ni nanoparticle-Gd-C is added to 25mL
In methanol, 1mLH is used2O and methanol dilution are to 150mL.Then the mixing of 150mL glycerine is added.Ultrasonic 30min, has been transferred to stirring
In the 500mL three-necked flasks of device.Add 10mL amino silicane coupling agents（AEAPS）, under 80-90 C after quick stirring 3h,
It is transferred out of product.Product is washed with deionized 3 times, and methanol washs 2 times (using Buchner funnel suction filtration).Vacuum drying.Need note
Meaning, suction filtration is due to there is glycerine, so slow, supernatant liquid, suction filtration mistake can be sucked with suction pipe in good time by taking out after a period of time
Journey about needs 6-8h.Obtained Aminosilylation Ni-Gd-C nano materials are finally dried in vacuo 12h.
Antibody modification：A certain amount of amido modified Ni-Gd-C nano-particles are weighed, the EDC and NHS of 2 times of quality is added,
Add 0.01M each 1mL of PBS.It is placed on blending instrument, 15 turns of rotating speed, reacts 45min.）The sample that soak time is arrived is carried out
10000r/min is centrifuged, time 5min, and the PBS for then adding 0.01M carries out washing centrifugation, is repeated 3 times.By the sample centrifuged
Product respectively add excessive Propiobacterium antibody, and 1h is coupled on blending instrument.The 10%BSA confining liquids 45min added after the completion of coupling.
After closing terminates, rotating speed 8000r/min centrifuges 5min, is centrifuged, is repeated 3 times with 0.01M PBS, then will be unnecessary
Antibody, BSA are washed away, and are eventually adding PBS and are resuspended.The Ni-Gd-C nano-particles of the resistance modification of preparation are immunological probe,
Be stored in 4 DEG C it is stand-by.
2）The culture medium sterilized in right amount is each added in a collection of polystyrene sample pipe, acne bar is inoculated with oxygen free condition
Bacterium, is separately added into after the Chinese medicine extract sealing for needing to screen, after culture a period of time, adds step 1）Prepared Ni-Gd-
C nano probe, is sufficiently mixed concussion reaction, if there is object bacteria in sample cell, first can be with Ni-Gd-C nano-probes surface
Antibody binding, so that Ni-Gd-C nano-probes are stayed in a liquid；If not there is object bacteria in sample cell, Ni-Gd-C receives
The antibody of rice detecting probe surface can be adsorbed gradually in polystyrene sample pipe surface.After reaction 1 hour, nuclear-magnetism is carried out to sample cell and is total to
Shake the measure in relaxation time（Shanghai knob steps NMR20 NMRs）.With the sterile medium liquid without vaccination target bacterium
For blank, the relaxation time spin-lattice relaxation time of the suspension measured has compared notable drop with spin spin relaxation time
Low, then explanation contains probe, illustrates there is object bacteria in sample cell, so that the bacteriostasis of side light Chinese medicine is not obvious, it is on the contrary
It is then anti-.The amount of probe and the decline of spin-lattice relaxation time and spin spin relaxation time are proportional, quantitative by mark-on
Probe and indirect quantification go out the amount of object bacteria.Illustrate that the Chinese medicine added does not have inhibitory action with the presence of object bacteria.It is on the contrary then anti-.
Embodiment
Example 2
Screening is to clostridium（Anaerobic bacteria）There is the Chinese medicine preparation of suppression or therapeutic action.
1）It is prepared by Ni nanoparticle-Gd-C immunological probes：
Ni-Gd-C nano-particles can be commercially（Such as the Chao Wei nano materials company in Shanghai）, can also bibliography
With physics and it is chemically synthesized.
Silicon dioxide coated Ni nanoparticle-Gd-C：47.5g sodium metasilicate is taken, is dissolved in deionized water in beaker, is adjusted with hydrochloric acid
Section pH value is 12-13.5.0g Ni nanoparticles-Gd-C is taken to be added in this beaker, mechanical agitation (using glass bar) 5min.By mixed liquor
Ultrasonic 30min, in good time stirring.85 C are warming up to, salt acid for adjusting pH value 6-7, generation precipitation is added dropwise.Spent in Magneto separate
Ion water washing is precipitated, and is washed 3-4 times.Then, precipitation is scattered in 250mL methanol.Above procedure is in triplicate, it is ensured that silicon
It is attached on Ni-Gd-C.
Amino containing silane Ni-Gd-C nano materials：Obtained silicon dioxide coated Ni nanoparticle-Gd-C is added to 25mL
In methanol, 1mLH is used2O and methanol dilution are to 150mL.Then the mixing of 150mL glycerine is added.Ultrasonic 30min, has been transferred to stirring
In the 500mL three-necked flasks of device.Add 10mL amino silicane coupling agents（AEAPS）, under 80-90 C after quick stirring 3h,
It is transferred out of product.Product is washed with deionized 3 times, and methanol washs 2 times (using Buchner funnel suction filtration).Vacuum drying.Need note
Meaning, suction filtration is due to there is glycerine, so slow, supernatant liquid, suction filtration mistake can be sucked with suction pipe in good time by taking out after a period of time
Journey about needs 6-8h.Obtained Aminosilylation Ni-Gd-C nano materials are finally dried in vacuo 12h.
Antibody modification：A certain amount of amido modified Ni-Gd-C nano-particles are weighed, the EDC and NHS of 2 times of quality is added,
Add 0.01M each 1mL of PBS.It is placed on blending instrument, 15 turns of rotating speed, reacts 45min.）The sample that soak time is arrived is carried out
10000r/min is centrifuged, time 5min, and the PBS for then adding 0.01M carries out washing centrifugation, is repeated 3 times.By the sample centrifuged
Product respectively add excessive clostridium antibody, and 1h is coupled on blending instrument.The 10%BSA confining liquids added after the completion of coupling
45min.After closing terminates, rotating speed 8000r/min centrifuges 5min, is centrifuged, is repeated 3 times with 0.01M PBS, then will
Unnecessary antibody, BSA are washed away, and are eventually adding PBS and are resuspended.The Ni-Gd-C nano-particles of the resistance modification of preparation are to exempt from
Epidemic disease probe, be stored in 4 DEG C it is stand-by.
2）The culture medium sterilized in right amount is each added in a collection of polystyrene sample pipe, fusiform bud is inoculated with oxygen free condition
Spore bacillus, is separately added into after the Chinese medicine extract sealing for needing to screen, after culture a period of time, adds step 1）Prepared
Ni-Gd-C nano-probes, are sufficiently mixed concussion reaction, if there is object bacteria in sample cell, first can be with Ni-Gd-C nano-probes
The antibody binding on surface, so that Ni-Gd-C nano-probes are stayed in a liquid.If not having object bacteria in sample cell, Ni-
The antibody on Gd-C nano-probes surface can be adsorbed gradually in polystyrene sample pipe surface.After reaction 1 hour, sample cell is carried out
The measure of NMR relaxation time（Shanghai knob steps NMR20 NMRs）.With the sterile culture without vaccination target bacterium
Base fluid body is blank, and the spin-lattice relaxation time of the suspension measured has compared notable drop with spin spin relaxation time
Low, then explanation contains probe, illustrates there is object bacteria in sample cell, so that the bacteriostasis of side light Chinese medicine is not obvious, it is on the contrary
It is then anti-.The amount of probe and the decline of spin-lattice relaxation time and spin spin relaxation time are proportional, quantitative by mark-on
Probe and indirect quantification go out the amount of object bacteria.Illustrate that the Chinese medicine added does not have inhibitory action with the presence of object bacteria.It is on the contrary then anti-.
Claims (3)
1. a kind of method that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on Ni-Gd-C nano-probes, its characterization step is as follows：
1）Target bacteria antibody is coated with the preparation of paramagnetic Ni-Gd-C nano-probes；
2）The culture medium of the anaerobic bacteria of sterilizing is added in polystyrene sample pipe, vaccination target anaerobic bacteria, addition needs screening
Chinese medicine preparation extract solution, after closed culture a period of time, step 1 is quantitatively adding with syringe）Prepared Ni-Gd-C nanometers
Probe, is sufficiently mixed concussion reaction for a period of time, if there is object bacteria in sample cell, the antibody on Ni-Gd-C nano-probes surface
It can be combined first with object bacteria, so that Ni-Gd-C nano-probes are stayed in a liquid；If not there is object bacteria in sample cell,
Then the antibody on Ni-Gd-C nano-probes surface can gradually adsorb and leave nutrient solution in polystyrene sample pipe surface；Now, it is right
Sample cell carries out the measure of NMR relaxation time；Using the sterile medium liquid without vaccination target bacterium as blank, survey
The relaxation time of the suspension obtained, then explanation contained probe compared to significantly reducing, so that have object bacteria in indirect proof sample,
The amount of probe and the decline in relaxation time are proportional, and by mark-on quantitative probe, indirect quantification goes out the amount of object bacteria；Sample cell
In there is no object bacteria to illustrate that the traditional Chinese medicine ingredients added have a fungistatic effect, it is on the contrary then anti-.
2. the method according to claim 1 that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on Ni-Gd-C nano-probes, its feature
It is that the Ni nanoparticle-Gd-C nano-probes are the Ni nanoparticle-Gd-C alloy materials with paramagnetic properties, nanometer particle size is less than 300
Nanometer.
3. the method according to claim 1 that anaerobic bacteria Inhibitors form Chinese Traditional Medicinal Herbs are screened based on Ni-Gd-C nano-probes, its feature
It is the change in final relaxation time of the detection evaluation method based on nuclear magnetic resonance technique of described object bacteria.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS61268187A (en) * | 1985-05-22 | 1986-11-27 | Kitasato Inst:The | Novel antibiotic substance sk-1071 and production thereof |
| EP0280894A1 (en) * | 1987-02-03 | 1988-09-07 | Mitsubishi Kasei Corporation | Novel antibiotic vermisporin, process for the production thereof and pharmaceutical composition comprising it as an active antibacterial agent |
| CN1808106A (en) * | 2005-01-21 | 2006-07-26 | 中国科学院上海药物研究所 | Method for screening acetylcholine esterase inhibitor by nuclear magnetic resonance |
-
2017
- 2017-04-10 CN CN201710228321.0A patent/CN107085004A/en not_active Withdrawn
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS61268187A (en) * | 1985-05-22 | 1986-11-27 | Kitasato Inst:The | Novel antibiotic substance sk-1071 and production thereof |
| EP0280894A1 (en) * | 1987-02-03 | 1988-09-07 | Mitsubishi Kasei Corporation | Novel antibiotic vermisporin, process for the production thereof and pharmaceutical composition comprising it as an active antibacterial agent |
| CN1808106A (en) * | 2005-01-21 | 2006-07-26 | 中国科学院上海药物研究所 | Method for screening acetylcholine esterase inhibitor by nuclear magnetic resonance |
Non-Patent Citations (1)
| Title |
|---|
| 申静 等: "基于NMR纳米探针的生物传感器用于快速检测沙门氏菌", 《食品工业科技》 * |
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Application publication date: 20170822 |