CN107525794A - Application of the polyvinyl alcohol-nano silver composite material in using Raman spectrum detection crystal violet - Google Patents
Application of the polyvinyl alcohol-nano silver composite material in using Raman spectrum detection crystal violet Download PDFInfo
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- CN107525794A CN107525794A CN201610467068.XA CN201610467068A CN107525794A CN 107525794 A CN107525794 A CN 107525794A CN 201610467068 A CN201610467068 A CN 201610467068A CN 107525794 A CN107525794 A CN 107525794A
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- polyvinyl alcohol
- nano silver
- composite material
- crystal violet
- gel
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/65—Raman scattering
- G01N21/658—Raman scattering enhancement Raman, e.g. surface plasmons
Abstract
The present invention discloses application of the polyvinyl alcohol-nano silver composite material in using Raman spectrum detection crystal violet; using sodium borohydride as reducing agent; the polyvinyl alcohol hydrogel of pattern and the controllable carrying nano silver of particle size is prepared using circulating frozen freezing process and infusion process; it is simple to operate; without complicated process equipment; polyvinyl alcohol can play a supporting role in protection as dispersion stabilizer to the nano-Ag particles with Electromagnetic enhancement.Crystal violet is detected with the polyvinyl alcohol hydrogel substrate of carrying nano silver, its detectable limit can reach 10‑12The order of magnitude, it is easy to use, it is simple to operate.
Description
Technical field
The invention belongs to the preparation of metal nano-macromolecule hydrogel composite and its answering in surface Raman enhancement field
With, be related to a kind of polyvinyl alcohol hydrogel of carrying nano silver preparation and its application as Raman substrate.
Background technology
Crystal violet is a kind of alkaline triphenylmethane cudbear, and solid powder glistens in dirty-green, is dissolved in water or ethanol
It is in afterwards purple, belongs to non-food raw material, it is extremely wide in bacteriology, cytology and histology etc. application, it is a kind of excellent
Biological stain.But also because its is cheap, easy to use, there is antibacterial isoreactivity, often misuse in aquatic products by illegal
Aquaculture.Crystal violet and its metabolite have compared with high toxicity, high residue, Long Term Contact or after taking, and are easily caused life
Body obtains cancer, distortion, mutation etc., there is very big harm to life entity and environment.Therefore this basic-dyeable fibre is residual
It is one of major issue of Food Safety Analysis to stay detection.
The method of detection crystal violet mainly has liquid chromatographic detection and Raman enhancing detection at present, wherein for Raman detection
Substrate is more, for example detects crystal violet using proof gold or fine silver nano material as substrate, but its cost is higher, operation is relative
It is cumbersome.In view of the shortcomings that many above, it is necessary to work out a kind of substrate that fast and convenient can detect crystal violet content.
The content of the invention
It is an object of the invention to overcome the deficiencies of the prior art and provide a kind of system of the polyvinyl alcohol hydrogel of carrying nano silver
The standby and its application as Raman substrate, using sodium borohydride as reducing agent, utilizes circulating frozen-freezing process and " dipping "
Method prepares the polyvinyl alcohol hydrogel of pattern and the controllable carrying nano silver of particle size, simple to operate, without complicated technology
Equipment, polyvinyl alcohol can play a supporting role in protection as dispersion stabilizer to the nano-Ag particles with Electromagnetic enhancement.
Crystal violet is detected with the polyvinyl alcohol hydrogel substrate of carrying nano silver, its detectable limit can reach 10-12The order of magnitude, user
Just, it is simple to operate.
The technical purpose of the present invention is achieved by following technical proposals：
Polyvinyl alcohol-nano silver composite material, is prepared as steps described below：
Step 1, the polyvinyl alcohol hydrogel of the presoma containing Nano Silver is prepared using circulating frozen-freezing process
Polyvinyl alcohol is dissolved in deionized water, the aqueous solution of silver nitrate is added into system and is uniformly dispersed, will uniformly be divided
The aqueous solution for dissipating polyvinyl alcohol and silver nitrate carries out circulating frozen-defrosting, to obtain the polyvinyl alcohol of the presoma containing Nano Silver
Hydrogel；
Specifically：Take 1.0~1.1g polyvinyl alcohol to be dissolved in 85~90 DEG C in 8ml deionized waters, stir 30min, then
Be cooled to room temperature, into system add 2ml concentration be 20~50mM silver nitrate solutiones, stirring 15min, after solution is taken
Go out centrifugation (400~600rpm, 3min) to drive the bubble in system out of.The syringe for being 1ml with capacity takes the above-mentioned institutes of 1ml
Solution is obtained, refrigerator is put and 12h is freezed at-18 DEG C, the lower lucifuge of 20-25 degrees Celsius of room temperature is thawed, freeze-thaw circulation 3~5
It is secondary, finally the gel after defrosting is pushed out in sample bottle, it is stand-by；
Step 2, use infusion process reduction gel in silver nitrate for Nano Silver
The polyvinyl alcohol hydrogel of the presoma containing Nano Silver prepared by step 1 is immersed in the aqueous solution of sodium borohydride, with
Silver ion is fully reacted and be reduced to Nano Silver, and be dispersed in hydrogel；
In step 2, reaction temperature is 20-25 degrees Celsius, soak time at least 24 hours, preferably 24-48 hours.
In step 2, the concentration ratio of sodium borohydride and silver nitrate is 3:1.
Step 3, the completely polyvinyl alcohol gel containing Nano Silver will be reduced in step 2 to be soaked in deionized water, every
8h changes a water, repeats 4~5 times, and to remove excess sodium borohydrate, rear taking-up gel carries out freezing with freeze drier and done
It is dry, that is, obtain the polyvinyl alcohol gel of dried carrying nano silver.
The polyvinyl alcohol gel of the polyvinyl alcohol-nano silver composite material, i.e. carrying nano silver that are prepared using the above method, is received
The size of rice Argent grain particle diameter is 5-50nm, and gel swelling is average up to 1000-2500%, and reaches the flat of balance
The equal used time is short, up to 5-10min.
Application of the above-mentioned polyvinyl alcohol-nano silver composite material in using Raman spectrum detection crystal violet, test limit are
10-12mol/l。
Gel is coated on slide and forms basalis by selection, i.e. SERS base materials, then will set SERS substrate materials
The slide of material is substantially immersed in the crystal violet aqueous solution respectively, select fully immersion so that base material fully with crystallization
Purple contact and/or absorption, while gel is reached swelling saturation state, such as slide is dipped into 5-10min in the aqueous solution.
Application in the Raman image research that crystal violet is distributed in animal body of above-mentioned polyvinyl alcohol-nano silver composite material.
Compared with prior art, the beneficial effects of the invention are as follows：(1) polyvinyl alcohol of the carrying nano silver prepared by the present invention
In hydrogel, polyvinyl alcohol can play support protection to the nano-Ag particles with Electromagnetic enhancement and make as dispersion stabilizer
With；(2) polyvinyl alcohol hydrogel of the carrying nano silver prepared by the present invention is as SERS substrates, the light with noble metal
Spectral property, and sample prepares simple, low cost and other advantages；(3) the polyvinyl alcohol water of the carrying nano silver prepared by the present invention
Gel includes network structure, has powerful water absorbing capacity, energy quick adsorption probe molecule, phase as SERS substrates
Than when traditional fine silver nano material substrate, test stability it is more preferable：(4) detection method of offer of the invention is with showing
The detection method for having crystal violet is compared, and has the characteristics of high sensitivity, its test limit can reach 10-12mol/l。
Brief description of the drawings
Silver nano-grain TEM photos in the polyvinyl alcohol hydrogel for the carrying nano silver that Fig. 1 is prepared for the present invention.
Fig. 2 is the SEM photograph of the polyvinyl alcohol hydrogel of carrying nano silver prepared by the present invention.
Fig. 3 is the polyvinyl alcohol hydrogel of carrying nano silver and the thermal gravimetric analysis curve of pure polyvinyl alcohol hydrogel prepared by the present invention.
Fig. 4 is the polyvinyl alcohol hydrogel of carrying nano silver and the ultraviolet-ray visible absorbing light of pure polyvinyl alcohol hydrogel prepared by the present invention
Spectrogram.
Fig. 5 is the polyvinyl alcohol hydrogel of carrying nano silver and the swellbility figure of pure polyvinyl alcohol hydrogel prepared by the present invention.
Fig. 6 is the SERS spectrograms that various concentrations crystal violet is detected in case study on implementation 4 of the present invention.
Fig. 7 is the white light photo and injection site schematic diagram of the nude mice used in the present invention.
Fig. 8 is the crystal violet Raman image picture at mouse bare subcutaneous injection position in the embodiment of the present invention.
Embodiment
Technical solution of the present invention is described in further detail with specific embodiment below in conjunction with the accompanying drawings, described specific reality
Apply example only the present invention is explained, be not intended to limit the invention.The type of polyvinyl alcohol 1799 (PVA1799)：
- CH2CHOH-n averages degree of polymerization 1750 ± 50, alcoholysis degree：98-99% (mol/mol)；SEM
(SEM):FEI NANO430 SEM；Transmission electron microscope (TEM)：FEI Tecnai G2 F20
Transmission electron microscope；Ultraviolet-uisible spectrophotometer (UV)：The UV, visible light spectrophotometrics of PerkinElmer Lambda 25
Meter；Raman spectrum (RS)：RENISHAW inVia reflex laser co-focusing micro-Raman spectroscopies；Electronic analysis day
It is flat：FA1004 (Shanghai).
Embodiment 1, foundation bibliography (YOSHIHIRO MORI, HITOSHI TOKURA, MASANORI YOSHIKAWA,
Properties of hydrogels synthesized by freezing and thawing aqueous polyvinyl
Alcohol solutions and their applications, JOURNAL OF MATERIALS SCIENCE 32 (1997)
The preparation of polyvinyl alcohol hydrogel 491-496) is carried out using circulating frozen-thawing mode
Take 1.1g polyvinyl alcohol to be dissolved in 90 DEG C in 8ml deionized waters, stir 30min, room temperature is subsequently cooled to, with appearance
The syringe measured as 1ml takes the above-mentioned resulting solutions of 1ml, puts refrigerator and 12h is freezed at -18 DEG C, and lucifuge is thawed at room temperature, cold
Gel after defrosting, is finally pushed out in sample bottle, is soaked in deionized water, every 8h by jelly-thaw cycles 4 times
A water is changed, is repeated 4 times.Gel is taken out afterwards to be freeze-dried with freeze drier, that is, obtains dried pure poly- second
Enol gel.
The preparation method of the polyvinyl alcohol hydrogel of the carrying nano silver of embodiment 2, comprises the following steps：
Step 1: the polyvinyl alcohol hydrogel of the presoma containing Nano Silver is prepared with circulating frozen-freezing process：Take the poly- second of 1.1g
Enol is dissolved in 8ml deionized waters in 90 DEG C, is stirred 30min, is subsequently cooled to room temperature, and it is dense that 2ml is added into system
Spend for 20mM silver nitrate solutiones, stir 15min, after solution taken out into centrifugation (500rpm, 3min) to drive system out of
In bubble.The syringe for being 1ml with capacity takes the above-mentioned resulting solutions of 1ml, puts refrigerator and freezes 12h at -18 DEG C,
Lucifuge is thawed at room temperature, and freeze-thaw is circulated 4 times, and finally the gel after defrosting is pushed out in sample bottle, stand-by；
Step 2: reduce the silver nitrate in gel with " dipping " method：The gel that 3mm is thick in step 1 is cut, by it
It is soaked in the sodium borohydride solution that 10ml concentration is 60mM, soaks 24h, makes its reaction complete；
Step 3: sample post-processes：The completely polyvinyl alcohol gel containing Nano Silver will be reduced in step 2 to be soaked in
In deionized water, a water is changed every 8h, is repeated 4 times, it is rear to take out gel freezing to remove excess sodium borohydrate
Drying machine is freeze-dried, that is, obtains the polyvinyl alcohol gel of dried carrying nano silver.
The preparation method of the polyvinyl alcohol hydrogel of the carrying nano silver of embodiment 3, compared with Example 2, difference is step
In one, it is 50mM silver nitrate solutiones to be changed to add 2ml concentration into system, and sodium borohydride concentration is in corresponding step 2
150mM。
The preparation method of the polyvinyl alcohol hydrogel of the carrying nano silver of embodiment 4, compared with Example 2, difference is step
In one, it is 100mM silver nitrate solutiones to be changed to add 2ml concentration into system, sodium borohydride concentration in corresponding step 2
For 300mM.
The silver nanoparticle restored in the step of (a), (b), (c) in Fig. 1 are respectively embodiment 2,3,4 two
Particle TEM photos, corresponding silver nano-grain particle diameter are 5-10nm, 20-25nm and 45-50nm respectively.With
The concentration of the silver-colored presoma and reducing agent of addition increases, the silver nanoparticle particle diameter increase restored in gel.
(a), (b), (c) in Fig. 2 are respectively the polyvinyl alcohol hydrogel of the carrying nano silver in embodiment 2,3,4
The SEM photograph of glue, (a) and (b) network size are bigger, and the network size in (c) is smaller.
(a) and (b) is respectively the polyvinyl alcohol hydrogel of carrying nano silver and pure polyethylene corresponding to embodiment 4 and 1 in Fig. 3
The thermal gravimetric analysis curve of alcohol hydrogel, PVA-Ag (AgNO in (a)3100mM) the thermal weight loss temperature range of hydrogel is
260~450 DEG C, and pure polyvinyl alcohol hydrogel heat decomposition temperature corresponding to (b) then starts from 210 DEG C, it was demonstrated that Ag nanometers
The addition of particle can aid in the heat endurance for improving PVA hydrogels.
(a) in Fig. 4, (b), (c), (d) are respectively the polyvinyl alcohol hydrogel of carrying nano silver in embodiment 4,3,2,1
The UV-visible absorption spectrum of glue and pure polyvinyl alcohol hydrogel, (a), (b), (c) occur bright near 405nm
The plasmon resonance absworption peak of aobvious Nano Silver, and the intensity of the absworption peak increases as silver nitrate concentration increases,
Show that nano Ag particles are successfully dispersed in hydrogel and the increase of nano Ag particles concentration, pure polyvinyl alcohol in (d)
Hydrogel is without obvious absorption peaks.
(a), (b), (c), (d) correspond to the polyvinyl alcohol water of carrying nano silver in embodiment 4,3,2,1 respectively in Fig. 5
The swellbility figure of gel and pure polyvinyl alcohol hydrogel, it is seen that the swellbility of the polyvinyl alcohol hydrogel of carrying nano silver is with nitre
The increase of sour silver concentration and increase, but when silver nitrate concentration maximum, i.e. the polyvinyl alcohol hydrogel of carrying nano silver corresponding to (a)
Required time is most long when glue reaches equilibrium swelling.It is placed in swellbility experiment using dried hydrogels in deionized water to absorption
Saturation state, swellbility=(weight-hydrogel dry of hydrogel saturation state)/hydrogel dry * 100%, swellbility
Average reachable 1000-2500%, and the average used time for reaching balance is shorter (up to 5-10min), it was demonstrated that water-setting of the present invention
Glue has loose structure and stronger water absorbing capacity.
Embodiment 5：The polyvinyl alcohol gel for the carrying nano silver being prepared by the use of embodiment 4 is as SERS substrates, with knot
Crystalviolet is Raman probe molecules, detects the content of crystal violet molecule.Enter by the polyvinyl alcohol gel of above-mentioned carrying nano silver
Row is in use, select that gel is coated on slide and forms basalis, i.e. SERS base materials, then will set SERS
The slide of base material is substantially immersed in 10 respectively-13、10-12、10-11、10-10、10-9、10-8、10-7Mol/l concentration
In the crystal violet aqueous solution, fully immersion is selected, so that base material is fully contacted and/or adsorbed with crystal violet, while makes to coagulate
Glue reaches swelling saturation state, such as slide is dipped into 5-10min in the aqueous solution.Naturally dry after taking-up slide,
Detection sample is made, is placed in Reinshaw laser Raman spectrometer, wherein：Frequency range is 2500cm-1—0cm-1, ripple
A length of 532nm, laser attenuation power are 0.01-0.05%, in a continuous mode, using the 2-5s time for exposure, are repeatedly adopted
Collection, complete the Raman detection of crystal violet.Fig. 6 is the SERS that various concentrations crystal violet is detected in case study on implementation 5 of the present invention
Spectrogram.As a result show, hydrogel of the invention can detect the characteristic peak of crystal violet well, when crystal violet concentration is
10-13During mol/l, 1620cm-1The peak at place is no longer obvious, i.e., is surveyed using the polyvinyl alcohol gel of carrying nano silver as SERS substrates
The test limit for obtaining crystal violet is 10-12mol/l。
Embodiment 6：Using the polyvinyl alcohol hydrogel of carrying nano silver as carrier, saturated crystallization purple molecular probe molecular solution is soaked
Swelling, is injected into nude mouse, observation imaging.
Step 1: by the polyvinyl alcohol hydrogel of carrying nano silver, 10 are completely immersed in-5Mol/l crystal violet solution to completely it is molten
Swollen saturation, after being dissolved under 90 degree, about 50 microlitres of nude mice by subcutaneous is injected into, to be measured, injection position such as Fig. 7 Block Diagrams
It is shown；
Step 2: by the injection crystal violet position of the nude mice sample of step 1 processing, Reinshaw laser Raman spectrometer is placed in
Under 5 times of object lens of microscope and focus on, wherein：The point centered on injection point, set constituency scope and step-length, Raman frequency shift
Scope is 2000cm-1—200cm-1, wavelength 633nm, laser attenuation power is 50%, in a static mode, is used
The 5s time for exposure, multipoint acquisition, the Raman Mapping detections of crystal violet are completed, and to 1621cm-1Place's peak intensity is drawn
Graceful imaging.
The white light photo and injection site schematic diagram and the knot at mouse bare subcutaneous injection position of nude mice during Fig. 7 and Fig. 8 is corresponded to respectively
Crystalviolet Raman image picture.Polyvinyl alcohol hydrogel detectable concentration by carrying nano silver is 10-5Mol/l crystal violet solutions exist
Distribution in nude mouse, and to 1621cm-1Place's Raman peak intensity does Raman image (such as Fig. 8).As a result show, in low power thing
Under mirror, the Raman response of crystal violet is still very sensitive, it is seen that has notable difference, i.e. this hair in injection site and peripheral region
Application (the Raman image that is in animal body distributed of the bright hydrogel in the Raman test that crystal violet is distributed in animal body
Research).
Exemplary description has been done to the present invention above, it should explanation, in the case where not departing from the core of the present invention,
Any simple deformation, modification or other skilled in the art can not spend the equivalent substitution of creative work to fall
Enter protection scope of the present invention.
Claims (7)
1. application of the polyvinyl alcohol-nano silver composite material in using Raman spectrum detection crystal violet, it is characterised in that
Polyvinyl alcohol-nano silver composite material is coated on slide and forms basalis, be i.e. SERS base materials, then will be set
The slide for putting SERS base materials is substantially immersed in the crystal violet aqueous solution respectively, fully immersion is selected, so that substrate
Material is fully contacted and/or adsorbed with crystal violet, while gel is reached swelling saturation state；
The size of nano-Ag particles particle diameter is 5-50nm in polyvinyl alcohol-nano silver composite material, and gel swelling is average
Up to 1000-2500%, and the average used time for reaching balance is short, up to 5-10min, is prepared as steps described below：
Step 1, the polyvinyl alcohol hydrogel of the presoma containing Nano Silver is prepared using circulating frozen-freezing process
Polyvinyl alcohol is dissolved in deionized water, the aqueous solution of silver nitrate is added into system and is uniformly dispersed, will uniformly be divided
The aqueous solution for dissipating polyvinyl alcohol and silver nitrate carries out circulating frozen-defrosting, to obtain the polyvinyl alcohol of the presoma containing Nano Silver
Hydrogel；
Step 2, use infusion process reduction gel in silver nitrate for Nano Silver
The polyvinyl alcohol hydrogel of the presoma containing Nano Silver prepared by step 1 is immersed in the aqueous solution of sodium borohydride, with
Silver ion is fully reacted and be reduced to Nano Silver, and be dispersed in hydrogel；
Step 3, the completely polyvinyl alcohol gel containing Nano Silver will be reduced in step 2 to be soaked in deionized water, every
8h changes a water, repeats 4~5 times, and to remove excess sodium borohydrate, rear taking-up gel carries out freezing with freeze drier and done
It is dry, that is, obtain the polyvinyl alcohol gel of dried carrying nano silver.
2. polyvinyl alcohol-nano silver composite material according to claim 1 is in using Raman spectrum detection crystal violet
Application, it is characterised in that in the preparation method of polyvinyl alcohol-nano silver composite material, in step 2, reaction
Temperature is 20-25 degrees Celsius, soak time at least 24 hours, preferably 24-48 hours.
3. polyvinyl alcohol-nano silver composite material according to claim 1 is in using Raman spectrum detection crystal violet
Application, it is characterised in that in the preparation method of polyvinyl alcohol-nano silver composite material, in step 2, boron hydrogen
The concentration ratio for changing sodium and silver nitrate is 3:1.
4. polyvinyl alcohol-nano silver composite material according to claim 1 is in using Raman spectrum detection crystal violet
Application, it is characterised in that test limit is 10-12mol/l。
5. the application in the Raman image research that crystal violet is distributed in animal body of polyvinyl alcohol-nano silver composite material, its
It is characterised by, the size of nano-Ag particles particle diameter is 5-50nm in polyvinyl alcohol-nano silver composite material, gel swelling
Degree is average up to 1000-2500%, and the average used time for reaching balance is short, up to 5-10min, enters as steps described below
It is prepared by row：
Step 1, the polyvinyl alcohol hydrogel of the presoma containing Nano Silver is prepared using circulating frozen-freezing process
Polyvinyl alcohol is dissolved in deionized water, the aqueous solution of silver nitrate is added into system and is uniformly dispersed, will uniformly be divided
The aqueous solution for dissipating polyvinyl alcohol and silver nitrate carries out circulating frozen-defrosting, to obtain the polyvinyl alcohol of the presoma containing Nano Silver
Hydrogel；
Step 2, use infusion process reduction gel in silver nitrate for Nano Silver
The polyvinyl alcohol hydrogel of the presoma containing Nano Silver prepared by step 1 is immersed in the aqueous solution of sodium borohydride, with
Silver ion is fully reacted and be reduced to Nano Silver, and be dispersed in hydrogel；
Step 3, the completely polyvinyl alcohol gel containing Nano Silver will be reduced in step 2 to be soaked in deionized water, every
8h changes a water, repeats 4~5 times, and to remove excess sodium borohydrate, rear taking-up gel carries out freezing with freeze drier and done
It is dry, that is, obtain the polyvinyl alcohol gel of dried carrying nano silver.
6. polyvinyl alcohol-nano silver composite material according to claim 5 Raman that crystal violet is distributed in animal body
Application in imaging research, it is characterised in that in the preparation method of polyvinyl alcohol-nano silver composite material, in step 2
In, reaction temperature is 20-25 degrees Celsius, soak time at least 24 hours, preferably 24-48 hours.
7. polyvinyl alcohol-nano silver composite material according to claim 5 Raman that crystal violet is distributed in animal body
Application in imaging research, it is characterised in that in the preparation method of polyvinyl alcohol-nano silver composite material, in step 2
In, the concentration ratio of sodium borohydride and silver nitrate is 3:1.
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| CN110857914A (en) * | 2018-08-23 | 2020-03-03 | 南京理工大学 | Poly (trifluoropropylmethylsiloxane)/silver composite surface enhanced Raman substrate and preparation method thereof |
| CN114062347A (en) * | 2021-11-12 | 2022-02-18 | 福州大学 | Flexible hydrogel SERS chip of aggregation-state silver nanoparticles |
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