CN110567940A - Silver-coated gold SERS substrate and preparation method thereof - Google Patents

Silver-coated gold SERS substrate and preparation method thereof Download PDF

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Publication number
CN110567940A
CN110567940A CN201911073807.7A CN201911073807A CN110567940A CN 110567940 A CN110567940 A CN 110567940A CN 201911073807 A CN201911073807 A CN 201911073807A CN 110567940 A CN110567940 A CN 110567940A
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silver
coated gold
gold
centrifuging
internal standard
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毕丽艳
白聪聪
郑庆印
马朋
杨影
王超云
张瑾锦
郑体花
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Binzhou Medical College
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Binzhou Medical College
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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y20/00—Nanooptics, e.g. quantum optics or photonic crystals
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y35/00—Methods or apparatus for measurement or analysis of nanostructures
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y40/00—Manufacture or treatment of nanostructures
    • 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

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  • Nanotechnology (AREA)
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  • Optics & Photonics (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)

Abstract

The invention relates to a silver-coated gold SERS substrate and a preparation method thereof. The method comprises the following steps: 0.2M CTAB and 5 x 10‑4HAuCl of M4Mixing and stirring, adding 3.8 x 10‑4NaHB of M4Stirring for 2min, and standing in water bath at 26-28 deg.C for 2 hr to obtain seeds; mixing CTAB and HAuCl4Sequentially mixing ascorbic acid and HCl, adding the seeds, uniformly stirring, and reacting for 4 hours in a water bath kettle at 26-28 ℃ to obtain gold nanorods; centrifuging and cleaning the gold nanorods, and mixing the gold nanorods with R6G to synthesize silver-coated gold of internal standard substances with different concentrations; and centrifuging and cleaning the silver-coated gold of the internal standard substances with different concentrations, then dripping the silver-coated gold on a clean silicon chip, and drying to obtain the SERS substrate. The invention has simple operationAnd the obtained product has high stability and is convenient to detect.

Description

Silver-coated gold SERS substrate and preparation method thereof
Technical Field
The invention relates to a silver-coated gold SERS substrate and a preparation method thereof.
Background
Generally, the detection method has higher requirements on detection of substances, namely high sensitivity and low detection limit. In view of the high efficiency and rapidness of the analysis method, people mostly select Surface-enhanced raman scattering (SERS for short) in many fields. The Raman probe can amplify or enhance signals of a substance to be detected by taking Raman molecules as probes. The normal Raman spectrum has the characteristics of small sample amount, no damage to sample components, strong specificity and the like, and has the advantages of not only having the advantages but also having stronger detection limit as low as 10 for the surface enhanced Raman spectrum-13And the detection range is wider. The SERS marking technology developed in the end of the 80's of the 20 th century on the basis of Surface Enhanced Raman Scattering (SERS) is a spectral marking technology for tracing an object to be detected using raman molecules as probes. The principle is that surface Raman active molecules are adsorbed on certain nano particles through adsorption so as to achieve the purpose of amplifying or enhancing Raman signals, and then the signals are read out as marking signals.
SERS probes generally refer to the labeling of some raman molecules on nanoparticles with SERS effect, which may be referred to as SERS-labeled nanoparticles. Wherein the molecules have definite common peak attribution and large Raman cross section, and a large conjugated system is contained in the molecules, or the resonance enhancement effect can be generated at certain excitation light wavelength.
In SERS detection of some substances with core-shell structures in recent years, the selection of an internal standard substance and the optimal concentration range of the internal standard substance are always the most time-consuming and exploration problems of the scientific research, the selection of a proper internal standard substance has no interference effect on signals of substances to be detected, the detection of the signals of the substances to be detected is also facilitated, the selection of the concentration is particularly critical, and therefore, the research and development of a probe for the proper internal standard substance is important.
Disclosure of Invention
Aiming at the defects of the prior art, the application provides a silver-coated gold SERS substrate and a preparation method thereof. The method is simple to operate, and the obtained product is high in stability and convenient to detect.
A preparation method of a silver-coated gold SERS substrate comprises the following steps:
(1) 0.2M CTAB and 5 x 10-4HAuCl of M4Mixing and stirring, adding 3.8 x 10-4NaHB of M4Stirring for 2min, and standing in water bath at 26-28 deg.C for 2 hr to obtain seeds; the CTAB and HAuCl4And NaHB4In a volume ratio of 25:25: 3;
25ml of 0.2M CTAB, 25ml of 4 x 10-3HAuCl of M4Sequentially mixing 0.4ml of 0.0788M ascorbic acid and 0.4ml of 1M HCl, then adding 0.14ml of the seeds, uniformly stirring, and reacting for 4 hours in a water bath kettle at 26-28 ℃ to obtain gold nanorods; (ii) a
(2) Centrifuging and cleaning the gold nanorods obtained in the step (1), and mixing the gold nanorods with R6G to synthesize silver-coated gold containing an internal standard substance;
(3) And (3) centrifuging and cleaning the silver-coated gold of the internal standard substance with different concentrations obtained in the step (2), then dripping the silver-coated gold on a clean silicon chip, and drying to obtain the SERS substrate.
Further, the step (2) centrifugally cleans the synthesized gold nanorods, and the specific conditions are as follows: centrifuging at 8000r/min for 5min, and removing supernatant.
Further, the step (2) of synthesizing the silver-coated gold containing the internal standard substance specifically comprises the following steps: centrifuging the synthesized gold nanorods, removing supernatant, adding a certain amount of R6G with different concentrations, mixing uniformly, standing in dark for 8-12h, and then, according to the volume ratio of CTAC to AgNO3VC =10:6:3, respectively adding 0.08M CTAC and 0.01M AgNO in sequence3And performing 0.1M VC to obtain the silver-coated gold containing the internal standard substance R6G.
further, the step (3) is to centrifuge and clean the silver-coated gold under the following conditions: centrifuging at 8000r/min for 5min, and removing supernatant; then, 1ml of triple distilled water was added thereto to carry out centrifugal washing under the same conditions.
The invention also relates to the silver-coated gold SERS substrate prepared by the preparation method.
Has the advantages that: the method is simple to operate, and the obtained product is high in stability and convenient to detect. The gold nanorods which have consistent length-diameter ratio and are uniformly arranged are synthesized, and the gold nanorods have high sensitivity and good repeatability; the generated gold nanorods have good stability and can be stored for more than 1 month at normal temperature; the synthesized silver-coated gold core-shell structure has uniform shape and size and good stability.
Drawings
Fig. 1 is a flow chart of a preparation process of a silver-coated gold SERS substrate according to example 1 of the present invention.
Fig. 2 is a scanning electron microscope image of a silver-coated gold SERS substrate according to example 1 of the present invention.
Fig. 3 is a transmission electron microscope image of a silver-coated gold SERS substrate according to example 1 of the present invention.
FIG. 4 shows an internal standard 1 x 10 according to example 1 of the present invention-2The silver-coated gold SERS detection spectrogram of R6G of M.
FIG. 5 shows an internal standard substance 5 x 10 according to example 2 of the present invention-3The silver-coated gold SERS detection spectrogram of R6G of M.
FIG. 6 is a graph of the SERS detection spectrum of silver coated gold of comparative example 1 of the present invention.
FIG. 7 is a graph of the SERS detection spectrum of silver-coated gold of comparative example 2 of the present invention.
FIG. 8 shows an internal standard 1 x 10 according to example 1 of the present invention-2R6G silver coated gold ultraviolet spectrum of M.
FIG. 9 shows an internal standard substance 5 x 10 according to example 2 of the present invention-3R6G silver coated gold ultraviolet spectrum of M.
FIG. 10 is a silver-coated gold UV spectrum of comparative example 3 of the present invention.
Detailed Description
Example 1
Fig. 1 is a flow chart for preparing a silver-coated gold SERS substrate, which illustrates the synthesized substances of the present application in a general manner by a simple flow chart, and for detecting the result, the steps of preparing the silver-coated gold SERS substrate are as follows:
1. Preparing seeds: 5ml of a 0.2M CTAB (cetyltrimethylammonium bromide) solution and 5ml of 5X 10-4HAuCl of M4The solution was added to a beaker with a stirrer in sequence, placed on a magnetic stirrer and then 0.6ml of NaHB was added4Solution (NaHB)4The solution was 0.01892g of NaHB4Shaking with 50ml ice-distilled water, and dissolving), changing the color of the solution from light yellow to brown yellow, stirring for 2min, and standing in 26-28 deg.C water bath for 2 hr to obtain seed.
2. Synthesizing gold nanorods: 25ml of 0.2M CTAB, 25ml of 4 x 10-3HAuCl of M40.4ml of 0.0788M VC (ascorbic acid) and 0.4ml of 1M HCl (hydrochloric acid) are sequentially added into a 100ml beaker to be uniformly mixed to obtain a growth solution, then 0.14ml of prepared seeds are added and uniformly stirred, and then the mixture is placed in a water bath kettle at 26-28 ℃ for 4 hours to obtain the gold nanorods. The gold nanorods have consistent length-diameter ratio and SERS activity.
3. Centrifuging gold nanorods 8000r/min for 5min, removing supernatant, and adding 0.02ml 1 × 10-3Mixing the R6G internal standard molecules of M uniformly, performing ultrasonic treatment for 3min, standing in dark for 8-12h, and then adding CTAC (cetyl trimethyl ammonium chloride) and AgNO in sequence3And (silver nitrate) and VC to obtain the silver-coated gold containing the internal standard substance R6G.
As an embodiment of the invention, CTAC, AgNO3, and VC are added in sequence, specifically: adding 0.2ml of 0.08M CTAC, then adding 1.6ml of CTAC, blowing uniformly, and then sequentially adding 1.2ml of 0.01M AgNO30.6ml of 0.1M VC is blown uniformly, the solution color is changed into light pink, and then the solution is placed in a water bath kettle at 65 ℃ for 4.5 hours, during which the solution color is changed from light pink into green and then changed into earthy yellow. (in the process of preparing silver-coated gold, the dissolution temperature of CTAC is required to be 55 ℃, the temperature of a centrifugal machine is strictly controlled during the centrifugation of gold nanorods, the temperature of the centrifugal machine is ensured to be not lower than 22 ℃, because the dissolution temperature of CTAB is 33 ℃ in the process of preparing the gold nanorods, and the CTAB can be caused by too low temperatureAnd the gold nano-rod is separated out from the solution, so that the morphology of the gold nano-rod is influenced, and the silver-coated gold can not be prepared by utilizing the gold nano-rod.
FIG. 2 is a scanning electron microscope photograph of gold-in-silver in example 1. Preparing a scanning electron microscope sample: centrifuging 1ml of synthesized silver-coated gold in two EP tubes at 8000r/min for 5min, removing supernatant, adding 1ml of triple distilled water, cleaning, centrifuging under the same conditions, removing supernatant, sucking about 6-10ul of concentrated solution with a pipette gun, dripping onto a silicon wafer, drying, and observing the appearance with a Scanning Electron Microscope (SEM) to obtain the figure 2.
FIG. 3 is a transmission electron microscope photograph of gold-in-silver in example 1. Preparation of transmission electron microscope samples: firstly, a silver-coated gold concentrated solution is prepared, the process is the same as the preparation process of the scanning electron microscope, then a proper amount of concentrated solution is absorbed by a liquid-transferring gun and dropped on a common carbon supporting film (copper net), after the concentrated solution is dried, the appearance of the silver-coated gold is represented by a Transmission Electron Microscope (TEM), and a graph 3 is obtained.
As can be seen from fig. 2 and 3, fig. 2 reflects the spatial three-dimensional structure of ag-au, and it can be seen from this figure that ag-au is cuboid, closely arranged, and shows a laminated distribution; FIG. 3 shows a two-dimensional structure of Ag-coated Au, which is rectangular, and the Au nanorods are clearly seen to be coated by the outer Ag, and the Ag-coated Au has uniform shape and size distribution.
Example 2
A method for preparing a silver-coated gold SERS substrate, which is the same as in example 1. Except that 0.02ml of 5 x 10 was added-3M R6G internal standard molecule.
Comparative example 1
A method for preparing a silver-coated gold SERS substrate, which is the same as in example 1. Except that no R6G internal standard molecule is added, the absorbance of the silver-coated gold is measured by an ultraviolet spectrophotometer, the concentration is obtained according to the Lambert beer law, and the concentration is diluted according to the corresponding multiple to obtain 1 x 10-2M silver coated gold.
Comparative example 2
A method for preparing a silver-coated gold SERS substrate, which is the same as in example 1. Except that no internal standard R6G was addedMeasuring the absorbance of the silver-coated gold by using an ultraviolet spectrophotometer, obtaining the concentration of the silver-coated gold according to the Lambert beer law, and diluting according to corresponding times to obtain 5 x 10-3M silver coated gold.
Comparative example 3
A preparation method of a silver-coated gold SERS substrate comprises the steps of firstly synthesizing gold nanorods according to the preparation process of example 1, centrifugally cleaning, then sequentially adding CTAC, AgNO3 and VC with the same concentration and volume, and placing in a 65 ℃ water bath kettle for 4.5 hours to obtain the silver-coated gold core-shell structure.
As shown in fig. 4 to 7, the SERS detection operation procedure for fig. 4 to 7 is as follows: a certain amount of silver-coated gold with different concentrations of R6G and silver-coated gold liquid with different concentrations are respectively sucked by a capillary, and data can be acquired by a Raman spectrometer and processed to obtain a spectrogram. The wavelength of the excitation light of the Raman spectrometer is 785 nm.
As can be seen from fig. 4 to 7, according to the data reflected by the following pictures, fig. 4 is compared with fig. 6, and fig. 5 is compared with fig. 7, and the two sets of compared SERS detection spectra show that the intensity of the raman spectrum of fig. 4 and 5 is obviously increased after the internal standard substance R6G is added; the silver coated gold Raman signal intensity measured in the absence of the internal standard R6G in the images of FIGS. 6 and 7 is low; compared with fig. 6 and 7, the intensity of the raman spectrum of fig. 4 and 5 is enhanced by nearly 10 times, which shows that the internal standard R6G can achieve the purpose of obvious raman signal amplification; from the data plots of fig. 4 and 5, 1 × 10 can be derived-2The intensity of the silver-coated gold raman spectrum of R6G of M was slightly stronger than that of the internal standard substance by 5 x 10-3Silver coated gold for R6G for M.
as shown in fig. 8 to 10, the uv spectrum data of fig. 8 to 10 can be obtained by adding 50ul of ag-coated gold prepared at different concentrations of R6G or ag-coated gold containing no internal standard molecule to a cuvette containing 3ml of triple distilled water, and then plotting.
As can be seen from fig. 8 to 10, the addition of the internal standard substance R6G obtained from the ultraviolet spectrum has one more peak than that of pure silver-coated gold, and the peak can visually reflect that the R6G is successfully coated by silver; according to the data in the following three graphs, it can be seen that the addition of the internal standard R6G does not affect the uv absorption of ag-au, and does not cause red shift and blue shift of wavelength, i.e. the addition of the internal standard R6G does not affect the vibration between covalent bonds in the ag-au structure at all.
The Raman molecules can be tightly adsorbed to the surface of the SERS substrate as much as possible, and have enough interaction to prevent desorption in certain processes; compared with other molecules, the Raman molecule of the application has a large Raman scattering cross section and strong Raman scattering capability.
The silver-coated gold core-shell structure has unique electronic and optical characteristics, and the shape, size and length-diameter ratio of the nano particles have a certain relation with the characteristics.
The application adopts Raman molecule R6G as interior label and carries out simple processing and can obtain higher sensitivity, and stability has good silver-coated gold SERS probe.
R6G as one of Raman molecules has good stability, and the SERS characteristic peak can be used as a good internal standard substance. The SERS probe taking R6G as a Raman molecule has the characteristics of high sensitivity, strong specificity, high physicochemical stability, good repeatability and the like, so that the SERS marking technology based on the surface enhanced Raman scattering is widely applied in the fields of national defense safety, environmental pollution, food safety, basic scientific research and the like, and is more frequently applied in the aspect of biomedicine.
The present invention may be embodied in other specific forms without departing from its spirit or essential characteristics. The above-described embodiments of the invention are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims, and not by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.

Claims (5)

1. A preparation method of a silver-coated gold SERS substrate is characterized by comprising the following steps:
(1) 0.2M CTAB and 5 x 10-4HAuCl of M4Mixing and stirring, adding 3.8 x 10-4NaHB of M4Stirring for 2min, and standing in water bath at 26-28 deg.C for 2 hr to obtain seeds; the CTAB and HAuCl4And NaHB4In a volume ratio of 25:25: 3;
25ml of 0.2M CTAB, 25ml of 4 x 10-3HAuCl of M4Sequentially mixing 0.4ml of 0.0788M ascorbic acid and 0.4ml of 1M HCl, then adding 0.14ml of the seeds, uniformly stirring, and reacting for 4 hours in a water bath kettle at 26-28 ℃ to obtain gold nanorods;
(2) Centrifuging and cleaning the gold nanorods obtained in the step (1), and mixing the gold nanorods with R6G to synthesize silver-coated gold containing an internal standard substance;
(3) And (3) centrifuging and cleaning the silver-coated gold of the internal standard substance with different concentrations obtained in the step (2), then dripping the silver-coated gold on a clean silicon chip, and drying to obtain the SERS substrate.
2. The preparation method according to claim 1, wherein the step (2) is to centrifugally wash the synthesized gold nanorods, under the specific conditions: centrifuging at 8000r/min for 5min, and removing supernatant.
3. The method according to claim 1, wherein the step (2) of synthesizing the silver-coated gold containing the internal standard substance comprises: centrifuging the synthesized gold nanorods, removing supernatant, adding a certain amount of R6G with different concentrations, mixing uniformly, standing in dark for 8-12h, and then, according to the volume ratio of CTAC to AgNO3VC =10:6:3, respectively adding 0.08M CTAC and 0.01M AgNO in sequence3And performing 0.1M VC to obtain the silver-coated gold containing the internal standard substance R6G.
4. the method according to claim 1, wherein the step (3) comprises centrifuging and washing the silver-coated gold under the following conditions: centrifuging at 8000r/min for 5min, and removing supernatant; then, 1ml of triple distilled water was added thereto to carry out centrifugal washing under the same conditions.
5. A silver-coated gold SERS substrate prepared by the preparation method according to any one of claims 1 to 4.
CN201911073807.7A 2019-11-06 2019-11-06 Silver-coated gold SERS substrate and preparation method thereof Pending CN110567940A (en)

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Citations (7)

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CN104677881A (en) * 2015-03-26 2015-06-03 厦门大学 Nanoparticles with core-shell structures and containing internal standard molecules and surface enhancement Raman quantitative detection method
CN106257271A (en) * 2015-06-19 2016-12-28 中国科学院宁波材料技术与工程研究所 A kind of composite based on surface enhanced raman spectroscopy technology and preparation method thereof
CN107328753A (en) * 2017-07-14 2017-11-07 厦门稀土材料研究所 The nanoparticle surface enhancing Raman spectrum quantitative analysis method of embedded internal standard molecule
CN107537438A (en) * 2017-08-23 2018-01-05 湖南大学 Magnetic composite nano material of graphene parcel and its preparation method and application
US20180067054A1 (en) * 2016-09-08 2018-03-08 Baker Hughes, A Ge Company, Llc Amine detection using surface enhanced raman spectroscopy with functionalized nanoparticles
US20190079014A1 (en) * 2016-03-24 2019-03-14 Nanyang Technological University Core-shell plasmonic nanogapped nanostructured material

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104614358A (en) * 2014-12-09 2015-05-13 临沂大学 Method for detecting thrombin based on Raman amplification of nanoparticle signal probe
CN104677881A (en) * 2015-03-26 2015-06-03 厦门大学 Nanoparticles with core-shell structures and containing internal standard molecules and surface enhancement Raman quantitative detection method
CN106257271A (en) * 2015-06-19 2016-12-28 中国科学院宁波材料技术与工程研究所 A kind of composite based on surface enhanced raman spectroscopy technology and preparation method thereof
US20190079014A1 (en) * 2016-03-24 2019-03-14 Nanyang Technological University Core-shell plasmonic nanogapped nanostructured material
US20180067054A1 (en) * 2016-09-08 2018-03-08 Baker Hughes, A Ge Company, Llc Amine detection using surface enhanced raman spectroscopy with functionalized nanoparticles
CN107328753A (en) * 2017-07-14 2017-11-07 厦门稀土材料研究所 The nanoparticle surface enhancing Raman spectrum quantitative analysis method of embedded internal standard molecule
CN107537438A (en) * 2017-08-23 2018-01-05 湖南大学 Magnetic composite nano material of graphene parcel and its preparation method and application

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Application publication date: 20191213