CN105727313A - Preparation method and application of carbon dots from beer - Google Patents
Preparation method and application of carbon dots from beer Download PDFInfo
- Publication number
- CN105727313A CN105727313A CN201410775191.9A CN201410775191A CN105727313A CN 105727313 A CN105727313 A CN 105727313A CN 201410775191 A CN201410775191 A CN 201410775191A CN 105727313 A CN105727313 A CN 105727313A
- Authority
- CN
- China
- Prior art keywords
- carbon point
- medicated beer
- preparation
- amycin
- carbon
- 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.)
- Pending
Links
Abstract
The invention discloses a preparation method and application of carbon dots from beer as a main raw material. In the method, the carbon dots are separated and purified through sephadex gel chromatography and then are collected. The carbon dots can directly be used in living cell fluorescent imaging, and in addition, the carbon dots are mixed and stirred with adriamycin in water to support the adriamycin onto the carbon dots, which can be used for killing tumor cells. The beer carbon dots are low in cost, low in toxicity and simple in method, has good water solubility when the adriamycin is supported thereon, and can be used in the fields of tumor fluorescent markers and treatment as a fluorescent medicine carrier.
Description
Technical field
The present invention relates to a kind of preparation method from medicated beer carbon point, and as fluorescence antineoplastic drug carrier, the application in tumor cell imaging and treatment.
Background technology
Cancer is a big disease of serious harm human health, and the annual whole world there are about 7,000,000 people and dies from cancer, and wherein 24% occurs in China.Treatment cancer mainly has three kinds of methods at present: operative therapy, chemotherapy and X-ray therapy.But the entrance of the chemotherapeutics of excess dosage is internal obvious side effect to the mankind.Therefore invention one can support cancer therapy drug, and fluorescence carrier that is that cancer carries out effectively treatment and that have no side effect is the great sciences problems of mankind's common concern.
Nano material is as a kind of new material in recent years, causes global concern and research boom.And fluorescent nano material receives much concern because it has the good advantage such as optical especially, it is made to have good application prospect (document 1:AlivisatosAPSemiconductorclusters in biomedicine field and nanocomposite optical device, nanocrystals, andquantumdots [J] .Science, 1996,271 (5251): 933-937).Semiconductor-quantum-point (such as CdSe) has superior light stability, water solublity, quantum yield high.But studies have found that recently, semiconductor-quantum-point, owing to containing heavy metal element, showing toxic and side effects in vivo, limits its application in biological medicine significantly.So finding a kind of fluorescent probe having no side effect is problem demanding prompt solution.Carbon quantum dot, is also called carbon point, is the newcomer of carbon nanometer family, and its average particle size distribution is at 1-10nm.Because its major part is containing only having C, H, O, three kinds of elements, compared with traditional quantum dot, it not only maintains original fluorescent stabilization and water solublity etc. a little, and there is obvious hypotoxicity and good biocompatibility and be easily functionalized (document 2:S.N.Baker, G.A.Baker, Luminescentcarbonnanodots:emergentnanolights.Angew.Chem., Int.Ed., 2010,49 (38), 6726-6744.).So fluorescent carbon point is considered as a kind of ideal fluorescent labeling for tumor and drug carrier material.
nullNearly ten years，The synthetic method of carbon point has increased significantly，Such as hydro-thermal method (document 3:B.Bourlinos,A.Stassinopoulos,D.Anglos,etal.SurfaceFunctionalizedCarbogenicQuantumDots.Small,2008,4(4):455-458.)，Oxidizing process (document 4:H.P.Liu,T.Ye,C.D.Mao,Fluorescentcarbonnanoparticlesderivedfromcandlesoot.Angew.Chem.,Int.Ed.,2007,46(34):6473-6475.)，Microwave assisting method (document 5:Q.L.Wang,H.Z.Zheng,Y.J.Long,L.Y.Zhang,etal.Microwave-hydrothermalsynthesisoffluorescentcarbondotsfromgraphiteoxide.Carbon,2011,49(9),3134-3140.)，But the condition generally synthesized is comparatively laborious，Required time is generally longer，Temperature is higher，It it is not the synthetic method of a kind of green.Daily bread and beverage are mainly made up of carbohydrate, can form carbon point in its course of processing.The carbon point that food and beverage are formed has no side effect, and has superior biocompatibility.Therefore in bio-imaging and medicine conveying, there is broader practice.
Summary of the invention
The present invention selects the fluorescence carrier as fluorescence cancer therapy drug using carbon naturally occurring in medicated beer first, the preparation method disclosing a kind of functional fluorescence carbon point pharmaceutical carrier, this medicated beer carbon is selected and can effectively be supported cancer therapy drug, and is applied to imaging and the treatment of tumor cell.
The technical scheme that the present invention takes is: the preparation method that the medicated beer carbon provided is selected is: concentrated by medicated beer, after removing insoluble impurities, by separating-purifying, collects hyperfluorescence component, dries to obtain carbon point；Being mixed with amycin 1000:1 1:10 in mass ratio by carbon point, be dissolved in deionized water, regulate pH value to 3-12, lucifuge stirs 0.5 72h, adds organic solvent, collects precipitation, obtains supporting amycin carbon point after volatile dry.
Described method for concentration is the combination of one or two or more kinds in rotary evaporation, lyophilization or spray drying.
Described process for separation and purification is the combination of a kind of in ultrafiltration, dialysis, ultracentrifugation, gel chromatography or any two and above method.
Described hyperfluorescence component is naturally occurring carbon quantum dot in medicated beer.
Described organic solvent is a kind of in methanol, ethanol, normal propyl alcohol, isopropanol, acetonitrile, acetone or any two or more than the two kinds mixture with arbitrary proportion composition, and the mass ratio of the addition of organic solvent and carbon point is 1:1-50:1.
Described carbon point, can as fluorescent drug carrier, apply in tumor fluorescent labeling and treatment, the carbon point supporting amycin arrives tumor locus, slow releasing medicine, uses the EPR effect of nanoparticle, strengthens oncotherapy effect, and in imaging, have better application, thereby increases and it is possible to the myocardial toxicity of amycin can be reduced.
Present invention have the advantage that
(1) raw material required for the present invention is easily purchased.
(2) carbon of the present invention is selected and is formed in the medicated beer course of processing, need not carry out specially
Synthesis.
(3) the medicated beer carbon point of the present invention has good fluorescence and biocompatibility.
(4) the medicated beer carbon point of the present invention has high safety.
(5) to support amycin antitumous effect obvious for the medicated beer carbon point of the present invention.
Accompanying drawing explanation
Fig. 1 is the transmission electron microscope photo of medicated beer carbon point.
Fig. 2 is the ultraviolet of medicated beer carbon point, fluorescence spectrum.
Fig. 3 is the fluorescence lifetime figure of medicated beer carbon point.
Fig. 4 is the XPS collection of illustrative plates of medicated beer carbon point.
Fig. 5 is the FTIR collection of illustrative plates of medicated beer carbon point.
Fig. 6 is the collection of illustrative plates of the XRD of medicated beer carbon point.
The ionic strength that Fig. 7 is medicated beer carbon point is stably schemed.
Fig. 8 is the cytotoxicity result of medicated beer carbon point.
Fig. 9 is the cytotoxicity result that medicated beer carbon point supports amycin.
Figure 10 is the cell marking figure of medicated beer carbon point.
Detailed description of the invention
Below by detailed description of the invention, the present invention will be further described.
Embodiment 1: the preparation of functional fluorescence carbon point and amycin support
(1) Tsingtao beer of about 100mL is steamed to about 5mL at 60 DEG C of condition backspins, pass sequentially through the membrane filtration of 0.45 μm, 0.22 μm, collect filtrate.Purifying with glucosan SephadexG-25 gel chromatography separation subsequently, observe under 340nm exciting light is assisted, collect the component of hyperfluorescence, lyophilization obtains carbon point.
(2) carbon point is mixed with the ratio of mass ratio 8:1 with amycin, and be dissolved in deionized water.With 1MNaOH aqueous solution, pH value is adjusted to 8, lucifuge stirring 24h.Adding the dehydrated alcohol of above-mentioned 8 times of volumes of mixed solution, high speed centrifugation (10000rpm, 10min) collects precipitation, obtains after drying supporting amycin carbon point.
Embodiment 2: the sign of medicated beer carbon point character
(1) form of medicated beer carbon point and size dimension
Fig. 1 is transmission electron microscope photo and the particle diameter cartogram of medicated beer carbon point, and result shows, the medicated beer carbon point size through separating-purifying is uniform, the regular one-tenth ball-shaped of form, add up medicated beer carbon point grain size concentrates on 2-3nm.
(2) fluorescence spectrum of medicated beer carbon point and ultraviolet spectrum characteristic
Fig. 2 is fluorescence spectrum figure and the ultraviolet spectra of medicated beer carbon point.In figure, there is obvious red shift along with excitation wavelength increases in the fluorescence spectrum of medicated beer carbon point, and the maximum excitation wavelength of coffee carbon point occurs in 340nm place.Ultraviolet spectrogram occurs in that the characteristic absorption peak of n → π * transition as seen at 270nm place.
(3) fluorescence lifetime of medicated beer carbon point
Fig. 3 is the fluorescence lifetime figure of medicated beer carbon point.The medicated beer carbon point aqueous solution of configuration 6mg/mL, excites under the exciting light of 376nm, and maximum emission peak is that 440nm launches, and records fluorescence lifetime.Finding that in experiment medicated beer carbon point is three index quenchers, the fluorescence lifetime being computed medicated beer carbon point is 4.79ns.
(4) the x-ray photoelectron power spectrum (XPS) of medicated beer carbon point characterizes
Fig. 4 is the collection of illustrative plates of medicated beer carbon point XPS.In figure, result shows to mainly contain two kinds of elements of C and O in medicated beer carbon point and containing minimal amount of N element.
(1) the Fourier's series infrared spectrum characterization of medicated beer carbon point
Fig. 5 is the infared spectrum of medicated beer carbon point.In figure, result shows the 3326cm of the medicated beer carbon point after purification-1Containing the stretching vibration peak of O-H and N-H, 2927cm-1There are the stretching vibration peak of saturated alkane, 1637cm-1There are the deformation vibration peak of N-H, 1044cm-1There is the stretching vibration peak of C-O.
(2) the XRD experiment of medicated beer carbon point
Fig. 6 is the collection of illustrative plates of the XRD of medicated beer carbon point, it is seen that 2theta=22 ° of place only has a steamed bread peak, composes according to XRD figure, illustrates that this carbon point is the structure with amorphous carbon.
(3) the pH stability experiment of medicated beer carbon point
Fig. 7 is the stable figure of the pH of medicated beer carbon point.The B-R buffer solution of differently configured pH (2 11), takes carbon point solution and is separately added in different pH solution (2mg/mL), survey fluorescence intensity with spectrofluorophotometer, take three meansigma methodss.Can be seen that in Fig. 8 that medicated beer carbon point fluorescence in acid condition is more stable, at pH=8 place, fluorescence intensity is decreased obviously, and under alkali condition, the comparatively acidic condition of fluorescence intensity is low.
Embodiment 3:
Cytotoxicity experiment
Select MCF-7 cell and the DMEM in high glucose culture medium of the hyclone containing 10%, add antibiotic and non essential amino acid.MCF-7 cell is inoculated in 96 orifice plates with 10000/hole, until adherent completely after (12h), discard former culture medium.After carbon point, the carbon point being supported with amycin and amycin being dissolved with culture medium respectively, then it is diluted to variable concentrations with culture medium, and joins in above-mentioned 96 orifice plates.After hatching 48h, measure appreciation rate with mtt assay.As shown in Figure 8, medicated beer carbon point (50mg/mL) under bigger concentration, MCF-7 cell survival rate remains to reach more than 80%, illustrates that medicated beer carbon point toxicity is relatively low.As it is shown in figure 9, MCF-7 cell is had bigger toxicity after supporting amycin by medicated beer carbon point.
Embodiment 4:
Medicated beer carbon point cell labeling experiment: MCF-7 cell forms single cell suspension after trypsinization, with 1 × 105The density of cells/mL is inoculated in culture dish, and adding 1ml medicated beer carbon point concentration in each culture medium is the culture medium of 20mg/ml, at 37 DEG C, and the CO of 5%224h is hatched under condition.Culture medium is gone with hypsokinesis, and with phosphate buffer eluting 2 times.Culture dish is placed under laser confocal microscope, excites under 405nm, 488nm wave band respectively, observe the cell marking situation of medicated beer carbon point.As shown in Figure 10, compared with matched group, obvious fluorescence can be sent through the cell of medicated beer carbon point labelling, under 405nm wave band excites, send obvious blue light, under 488nm wave band excites, send obvious green glow, illustrate that medicated beer carbon point can as good fluorescent material for cell fluorescence imaging.
Claims (6)
1. the preparation method from the carbon point of medicated beer, it is characterised in that: medicated beer is concentrated, after removing insoluble impurities, by separating-purifying, collects hyperfluorescence component, dry to obtain carbon point；Being mixed with amycin 1000:1 1:10 in mass ratio by carbon point, be dissolved in deionized water, regulate pH value to 3-12, lucifuge stirs 0.5 72h, adds organic solvent, collects precipitation, obtains supporting amycin carbon point after volatile dry.
2. preparation method according to claim 1, it is characterised in that: described method for concentration is the combination of one or two or more kinds in rotary evaporation, lyophilization or spray drying.
3. preparation method according to claim 1, it is characterised in that: described process for separation and purification is the combination of a kind of in ultrafiltration, dialysis, ultracentrifugation, gel chromatography or any two and above method.
4. preparation method according to claim 1, it is characterised in that: described hyperfluorescence component is naturally occurring carbon quantum dot in medicated beer.
5. preparation method according to claim 1, it is characterized in that: it is characterized in that: described organic solvent is a kind of in methanol, ethanol, normal propyl alcohol, isopropanol, acetonitrile, acetone or any two or more than the two kinds mixture with arbitrary proportion composition, the mass ratio of the addition of organic solvent and carbon point is 1:1-50:1.
6. the application of the carbon point described in a claim 1, it is characterized in that: described carbon point, can as fluorescent drug carrier, applying in tumor fluorescent labeling and treatment, the carbon point supporting amycin arrives tumor locus, slow releasing medicine, use the EPR effect of nanoparticle, strengthen oncotherapy effect, and in imaging, have better application, thereby increases and it is possible to the myocardial toxicity of amycin can be reduced.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410775191.9A CN105727313A (en) | 2014-12-12 | 2014-12-12 | Preparation method and application of carbon dots from beer |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410775191.9A CN105727313A (en) | 2014-12-12 | 2014-12-12 | Preparation method and application of carbon dots from beer |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105727313A true CN105727313A (en) | 2016-07-06 |
Family
ID=56240952
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410775191.9A Pending CN105727313A (en) | 2014-12-12 | 2014-12-12 | Preparation method and application of carbon dots from beer |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105727313A (en) |
Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107115319A (en) * | 2017-04-05 | 2017-09-01 | 汪涛 | A kind of biological safety carbon quantum dot load adriamycin complex and its preparation method and application |
| CN107603610A (en) * | 2017-09-14 | 2018-01-19 | 大连工业大学 | Preparation method from mature vinegar carbon nano-particles |
| CN107663453A (en) * | 2016-07-28 | 2018-02-06 | 东北林业大学 | A kind of preparation method of lignin fluorescent carbon nano-particles |
| CN108084996A (en) * | 2017-12-28 | 2018-05-29 | 大连工业大学 | The method that extraction prepares fluorescent carbon quantum dot from beer |
| CN108273071A (en) * | 2018-01-26 | 2018-07-13 | 大连工业大学 | A kind of food-borne fluorescence nano grain and its preparation method and application |
| CN108379592A (en) * | 2018-04-10 | 2018-08-10 | 中山大学附属第五医院 | A kind of fluorescent carbon point support adriamycin FRET combination drug carriers and the preparation method and application thereof through PEI modifications |
| CN108837156A (en) * | 2018-06-21 | 2018-11-20 | 河北师范大学 | A kind of preparation method of carbon dots medicine-carried system |
| CN109467074A (en) * | 2018-12-13 | 2019-03-15 | 南京师范大学 | A kind of carbon quantum dot and the preparation method and application thereof improving adriamycin anti-cancer effectiveness |
| CN109678141A (en) * | 2019-03-04 | 2019-04-26 | 上海交通大学 | Graphene quantum dot separation method based on cross-linked dextran gel column |
| WO2021169744A1 (en) * | 2020-02-28 | 2021-09-02 | 复旦大学 | Fluorescent carbon quantum dot, and preparation method therefor and application thereof in preparation of antitumor drug sensitizer |
| CN113548656A (en) * | 2020-06-16 | 2021-10-26 | 哈尔滨成程生命与物质研究所 | Carbon dots with anticancer bioactivity and preparation method thereof |
| CN113999675A (en) * | 2021-11-29 | 2022-02-01 | 江苏科技大学 | Cell-derived fluorescent carbon nanosheet and preparation method and application thereof |
-
2014
- 2014-12-12 CN CN201410775191.9A patent/CN105727313A/en active Pending
Non-Patent Citations (5)
| Title |
|---|
| CHENGKUN JIANG等: ""PresenceofphotoluminescentcarbondotsinNescafes original instant coffee:Applicationstobioimaging"", 《TALANTA》 * |
| JING TANG等: ""Carbon Nanodots Featuring Effi cient FRET for Real-Time Monitoring of Drug Delivery and Two-Photon Imaging"", 《ADVANCED MATERIALS》 * |
| SO YOUNG PARK等: ""Photoluminescent Green Carbon Nanodots from Food-Waste-Derived Sources: Large-Scale Synthesis, Properties, and Biomedical applications"", 《ACS APPLIED MATERIALS&INTERFACES》 * |
| 刘玉梅等: ""烘烤温度对啤酒花品质的影响"", 《酿洒科技》 * |
| 陈霞等: ""焙烤麦芽的麦汁和啤酒中一个稳定自由基的表征"", 《啤酒科技》 * |
Cited By (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107663453B (en) * | 2016-07-28 | 2022-01-11 | 东北林业大学 | Preparation method of lignin fluorescent carbon nanoparticles |
| CN107663453A (en) * | 2016-07-28 | 2018-02-06 | 东北林业大学 | A kind of preparation method of lignin fluorescent carbon nano-particles |
| CN107115319A (en) * | 2017-04-05 | 2017-09-01 | 汪涛 | A kind of biological safety carbon quantum dot load adriamycin complex and its preparation method and application |
| CN107603610A (en) * | 2017-09-14 | 2018-01-19 | 大连工业大学 | Preparation method from mature vinegar carbon nano-particles |
| CN108084996A (en) * | 2017-12-28 | 2018-05-29 | 大连工业大学 | The method that extraction prepares fluorescent carbon quantum dot from beer |
| CN108273071A (en) * | 2018-01-26 | 2018-07-13 | 大连工业大学 | A kind of food-borne fluorescence nano grain and its preparation method and application |
| CN108379592A (en) * | 2018-04-10 | 2018-08-10 | 中山大学附属第五医院 | A kind of fluorescent carbon point support adriamycin FRET combination drug carriers and the preparation method and application thereof through PEI modifications |
| CN108837156A (en) * | 2018-06-21 | 2018-11-20 | 河北师范大学 | A kind of preparation method of carbon dots medicine-carried system |
| CN108837156B (en) * | 2018-06-21 | 2021-08-17 | 河北师范大学 | Preparation method of carbon dot drug-loading system |
| CN109467074A (en) * | 2018-12-13 | 2019-03-15 | 南京师范大学 | A kind of carbon quantum dot and the preparation method and application thereof improving adriamycin anti-cancer effectiveness |
| CN109678141A (en) * | 2019-03-04 | 2019-04-26 | 上海交通大学 | Graphene quantum dot separation method based on cross-linked dextran gel column |
| WO2021169744A1 (en) * | 2020-02-28 | 2021-09-02 | 复旦大学 | Fluorescent carbon quantum dot, and preparation method therefor and application thereof in preparation of antitumor drug sensitizer |
| CN113548656A (en) * | 2020-06-16 | 2021-10-26 | 哈尔滨成程生命与物质研究所 | Carbon dots with anticancer bioactivity and preparation method thereof |
| CN113548656B (en) * | 2020-06-16 | 2023-02-21 | 哈尔滨成程生命与物质研究所 | Carbon dots with anticancer bioactivity and preparation method thereof |
| CN113999675A (en) * | 2021-11-29 | 2022-02-01 | 江苏科技大学 | Cell-derived fluorescent carbon nanosheet and preparation method and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105727313A (en) | Preparation method and application of carbon dots from beer | |
| Qin et al. | pH-responsive polymer-stabilized ZIF-8 nanocomposites for fluorescence and magnetic resonance dual-modal imaging-guided chemo-/photodynamic combinational cancer therapy | |
| Song et al. | Bioimaging based on fluorescent carbon dots | |
| Guan et al. | Fullerene/photosensitizer nanovesicles as highly efficient and clearable phototheranostics with enhanced tumor accumulation for cancer therapy | |
| Yan et al. | Enhanced fluorescence imaging guided photodynamic therapy of sinoporphyrin sodium loaded graphene oxide | |
| Jeong et al. | Fluorescent carbon nanoparticles derived from natural materials of mango fruit for bio-imaging probes | |
| Wang et al. | Drug delivery with upconversion nanoparticles for multi-functional targeted cancer cell imaging and therapy | |
| Guo et al. | A graphene oxide-based switch-on fluorescent probe for glutathione detection and cancer diagnosis | |
| CN110201163B (en) | Hyaluronic acid and polydopamine modified drug-loaded mesoporous titanium dioxide nanoparticle | |
| Ko et al. | In vivo imaging of tumour bearing near-infrared fluorescence-emitting carbon nanodots derived from tire soot | |
| Sun et al. | The cost-effective preparation of green fluorescent carbon dots for bioimaging and enhanced intracellular drug delivery | |
| Li et al. | Core-satellite metal-organic framework@ upconversion nanoparticle superstructures via electrostatic self-assembly for efficient photodynamic theranostics | |
| Mukherjee et al. | Tunable luminescent carbon nanospheres with well‐defined nanoscale chemistry for synchronized imaging and therapy | |
| Chen et al. | NIR light controlled release of caged hydrogen sulfide based on upconversion nanoparticles | |
| Ruan et al. | A simple one-step method for preparation of fluorescent carbon nanospheres and the potential application in cell organelles imaging | |
| Zhang et al. | Perylenediimide chromophore as an efficient photothermal agent for cancer therapy | |
| CN105754593A (en) | Hollow fluorescent carbon quantum dots as well as preparation method and application thereof | |
| Ren et al. | Near-infrared fluorescent carbon dots encapsulated liposomes as multifunctional nano-carrier and tracer of the anticancer agent cinobufagin in vivo and in vitro | |
| CN103566381A (en) | Multifunctional magnetic nano particle and preparation method thereof | |
| CN104083771A (en) | Fluorescence resonance energy transfer-based tumor imaging and therapeutic targeting system and construction method thereof | |
| Jin et al. | Targeted polypyrrole nanoparticles for the identification and treatment of hepatocellular carcinoma | |
| Sun et al. | Targeted and imaging-guided in vivo photodynamic therapy for tumors using dual-function, aggregation-induced emission nanoparticles | |
| Nair et al. | Gold nanorods decorated with a cancer drug for multimodal imaging and therapy | |
| CN110408047B (en) | Nano coordination polymer and preparation method and application thereof | |
| CN107158410B (en) | Folic acid-chitosan-Cy 7 polymer with tumor targeting property and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20160706 |