CN106124476A - Based on surface enhanced raman spectroscopy and the glucose sensing approach of bi-molecular probe - Google Patents
Based on surface enhanced raman spectroscopy and the glucose sensing approach of bi-molecular probe Download PDFInfo
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- CN106124476A CN106124476A CN201610450103.7A CN201610450103A CN106124476A CN 106124476 A CN106124476 A CN 106124476A CN 201610450103 A CN201610450103 A CN 201610450103A CN 106124476 A CN106124476 A CN 106124476A
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- glucose
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- molecular probe
- sers
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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 provides a kind of based on surface enhanced raman spectroscopy (SERS) and the glucose sensing approach of bi-molecular probe, a species specificity is provided strong for glucose detection, highly sensitive means, cannot be only used for direct blood sugar test, it may also be used for the detection of the human sample that the glucose contents such as urine, saliva and perspiration are relatively low.SERS technology realizes the glucose detection of high sensitivity, non-invasive manner, the gold silver core-shell nanometer rod that have employed the character such as shape, size highly controllable prepares SERS active-substrate, the phasmon wavelength of substrate accurately can be regulated and resonate to incident illumination, thus reach the highest Raman enhancer, maximized raising detection sensitivity.Have employed 4 mercaptophenyl boronic acid and 4 cyanophenylboronic acid bi-molecular probe technology, it is achieved thereby that the specificity detection of glucose, the Characteristic Raman peak of 4 cyanophenylboronic acids is positioned at biological quiescent region, thus effectively prevent the interference of other endogenous biological molecule, it is ensured that the specificity of detection and accuracy.
Description
Technical field
The present invention relates to blood sugar test field, more particularly, to one based on surface enhanced raman spectroscopy and bimolecular
The glucose quantitation detection method of probe.
Background technology
Commonly used with in family blood-sugar detecting instrument clinically is that blood directly detects, and blood collection can be given and suffer from frequently
Person brings discomfort.Examination of sugar in urine is the non-invasive simplest method of diabetes primary dcreening operation, almost without glucose in Healthy People urine,
Only when blood sugar concentration is higher than renal glucose threshold (8.88 ~ 9.99 mmol/l), urine just there will be glucose, and concentration and blood
Sugar concentration has preferable dependency, but is affected by many factors because of screening test at present, it is impossible to accurate reaction blood glucose
Concentration, thus its reference value is limited.Study confirmation, for individuality, in the human body body fluid such as saliva, perspiration before the meal
Concentration of glucose also has good dependency with blood sugar concentration, can be used for indirect reaction blood sugar concentration, if it can accurately be measured
In glucose content, in fields such as diabetes early screening, blood sugar test, will there is huge using value, but due to its Portugal
Grape sugar concentration is relatively low, generally 2*10-2 mmol/l ~ 2.4*10-1Mmo/l), there is no the commercially available blood sugar test instrument of method accurate
Really detect.Therefore, detection sensitivity glucose quantitation detection method high, fireballing will bring new contract for clinical practice
Machine.
Surface enhanced raman spectroscopy (SERS) detection technique is a kind of noinvasive, highly sensitive finger-print type spectral detection skill
Art, can provide the information about the structure of matter on a molecular scale, and the enhancing to Raman signal may be up to 1014Times.Therefore, SERS
Detection technique is with a wide range of applications in biomedical sensing detection field.Chinese patent CN103411949A discloses one
Planting the method utilizing SERS technology indirect detection glucose in serum content, the method is at glucoseoxidase by glucose
Effect under produce hydrogen peroxide molecule, in the presence of produced hydrogen peroxide molecule, divided by oxidation state chromogenic substrate
The intensity size of the surface-enhanced Raman signals of son obtains the glucose content in serum indirectly.Demonstrate surface-enhanced Raman
Technology feasibility in glucose quantitation detects.
Malini Olivo et al. (journal article, Biosensors and Bioelectronics, 56 (2014)
186– 191;J. Am. Chem. Soc. 2013,135,18028 18031) propose to utilize sandwich style bimolecular to visit
Pin technology realizes the specificity detection of glucose, utilizes 4-Mercaptobenzoic acid-three osmium Carbonyl Clusters conjugate or alkynyl phenylboric acid
SERS detection is carried out with the use of the selective glucose molecule that catches, to realize the high selection of glucose with 4-mercaptophenyl boronic acid
Property detection by quantitative.But, owing to the method is by spin on polymers microsphere and sputtering silver, gold thin film, slide is prepared relatively
For uniform nanostructured SERS active-substrate, the stabilization time of substrate is shorter, and due to the phasmon of its nanostructured
Wavelength cannot regulate, and causes its detection sensitivity limited, and these are all unfavorable for clinical practice.
Summary of the invention
The present invention is the defect overcoming the detection sensitivity described in above-mentioned prior art limited, it is provided that a species specificity is strong,
Highly sensitive, noninvasive based on surface enhanced raman spectroscopy with the glucose sensing approach of bi-molecular probe.
For solving above-mentioned technical problem, technical scheme is as follows:
Based on surface enhanced raman spectroscopy (SERS) and the glucose sensing approach of bi-molecular probe, described method includes following step
Rapid:
S1: by the coin race metal nano-rod uniform deposition for preparing at carrier surface, be prepared as surface enhanced raman spectroscopy
(SERS) active substrate, the size of regulation coin race metal nano-rod and transverse and longitudinal ratio, make the phasmon wavelength of SERS active-substrate
Resonate with incident illumination；
S2: by primary glucoreceptor molecular modification SERS active-substrate so that it is be fixed in SERS active-substrate；
SERS active-substrate is soaked in the Glucose standards aqueous solution of variable concentrations, the glucose in solution is captured
In SERS active-substrate；
Adding two grades of glucoreceptor molecules in SERS active-substrate, two grades of glucoreceptor molecules are selective and in substrate
Glucose molecule covalent bond；
Carry out SERS spectra test, demarcate the standard of concentration known with the Raman signatures peak intensity of two grades of glucoreceptor molecules
D/W, sets up quantitative analysis standard curve；
After S3: gather the sample to be tested that concentration of glucose is unknown, suitably dilution, carry out processing and SERS light by the method for step S2
Spectrum test, substitutes into, by the Raman signatures peak intensity of obtain two grades of Fructus Vitis viniferae acceptor molecules, the quantitative analysis standard that step S2 is set up
Curve, obtains the glucose concentration value of sample to be tested.
In the preferred scheme of one, described coin race metal nano-rod is Jin-silver core-shell nanometer rod.
In the preferred scheme of one, described carrier is coverslip.
In the preferred scheme of one, described carrier is filter paper or filter membrane.
In the preferred scheme of one, the instrument that SERS spectra test uses is Raman spectrometer, the wavelength of its exciting light
Scope is 400 ~ 1000 nm.
In the preferred scheme of one, described sample to be tested includes blood, urine, saliva, perspiration.
In the preferred scheme of one, described primary glucoreceptor molecule uses 4-mercaptophenyl boronic acid molecule, and it is fixed
In SERS active-substrate.
In the preferred scheme of one, described two grades of glucoreceptor molecules use 4-cyanophenylboronic acid, and it is lived with SERS
Property suprabasil glucose molecule combine.
In the preferred scheme of one, use the cyano group raman characteristic peak of 4-cyanophenylboronic acid that concentration of glucose is carried out fixed
Amount detection.
Compared with prior art, technical solution of the present invention provides the benefit that: the present invention provides a kind of based on surface enhanced
Raman scattering and the glucose sensing approach of bi-molecular probe, provide a species specificity for glucose detection strong, highly sensitive
Means, cannot be only used for blood sugar test, it may also be used for the human sample that the glucose contents such as urine, saliva and perspiration are relatively low
Detection.SERS technology realizes the glucose detection of high sensitivity, non-invasive manner, have employed the character such as shape, size highly controllable
Jin-silver core-shell nanometer rod prepare SERS active-substrate, the phasmon wavelength of substrate accurately can be regulated to incident illumination altogether
Shake, thus reach the highest Raman enhancer, maximized raising detection sensitivity.Have employed 4-mercaptophenyl boronic acid and 4-cyanogen
Base phenylboric acid bi-molecular probe technology, it is achieved thereby that the specificity detection of glucose, the cyano group feature of 4-cyanophenylboronic acid is drawn
Man Feng is positioned at biological quiescent region, thus effectively prevent the interference of other endogenous biological molecule, it is ensured that the specificity of detection
And accuracy.
Accompanying drawing explanation
Fig. 1 is the schematic diagram of glucose sensing approach of the present invention.
Fig. 2 is the schematic diagram of Jin-silver core-shell nanometer rod SERS substrate.
Fig. 3 is the schematic diagram of glucose detection.
Detailed description of the invention
Accompanying drawing being merely cited for property explanation, it is impossible to be interpreted as the restriction to this patent；
In order to the present embodiment is more preferably described, some parts of accompanying drawing have omission, zoom in or out, and do not represent the chi of actual product
Very little；
To those skilled in the art, in accompanying drawing, some known features and explanation thereof may will be understood by omission.
With embodiment, technical scheme is described further below in conjunction with the accompanying drawings.
Embodiment 1
As Figure 1-3, based on surface enhanced raman spectroscopy and the glucose sensing approach of bi-molecular probe, described method includes
Following steps:
S1: Jin-silver core-shell nanometer rod (the silver gold-covered nano rod) uniform deposition prepared, on coverslip surface, is prepared as SERS
Active substrate, the size of regulation Jin-silver core-shell nanometer rod and transverse and longitudinal ratio, make the phasmon wavelength of SERS active-substrate with incident
Photoresonance；
S2: modify SERS active-substrate with 4-mercaptophenyl boronic acid, make 4-mercaptophenyl boronic acid be fixed in SERS active-substrate, as
Primary glucoreceptor；
SERS active-substrate is soaked in the Glucose standards aqueous solution of variable concentrations, the glucose in solution is captured
In SERS active-substrate；
In SERS active-substrate add two grades of glucoreceptor 4-cyanophenylboronic acids, 4-cyanophenylboronic acid molecule optionally with
Glucose molecule covalent bond in substrate；
Carry out SERS spectra test, demarcate the standard glucose of concentration known with the Raman signatures peak intensity of 4-cyanophenylboronic acid
Aqueous solution, sets up quantitative analysis standard curve；
S3: use the Raman spectrometer that wave-length coverage is 400 ~ 1000 nm of exciting light, gathers the blood that concentration of glucose is unknown
After liquid, urine, saliva or perspiration sample, suitably dilution, carry out processing and SERS spectra test by the method for step S2, will obtain
The Raman signatures peak intensity of 4-cyanophenylboronic acid substitute into the quantitative analysis standard curve set up of step S2, obtain sample to be tested
Glucose concentration value.
The present invention provides a kind of based on surface enhanced raman spectroscopy with the glucose sensing approach of bi-molecular probe, for Fructus Vitis viniferae
It is strong that sugar detection provides a species specificity, and highly sensitive means cannot be only used for blood sugar test, it may also be used for urine, saliva
The detection of the human sample relatively low with glucose contents such as perspiration.SERS technology realizes the glucose of high sensitivity, non-invasive manner
Detection, have employed the highly controllable Jin of the character such as shape, size-silver core-shell nanometer rod and prepares SERS active-substrate, can be by substrate
Phasmon wavelength accurately regulate and resonate to incident illumination, thus reach the highest Raman enhancer, maximized raising
Detection sensitivity.Have employed 4-mercaptophenyl boronic acid and 4-cyanophenylboronic acid bi-molecular probe technology, it is achieved thereby that glucose
Specificity detects, and the Characteristic Raman peak of 4-cyanophenylboronic acid is positioned at biological quiescent region, thus it is raw to effectively prevent other endogenouss
The interference of thing molecule, it is ensured that the specificity of detection and accuracy.
The corresponding same or analogous parts of same or analogous label；
Term the being merely cited for property explanation of position relationship described in accompanying drawing, it is impossible to be interpreted as the restriction to this patent；
Obviously, the above embodiment of the present invention is only for clearly demonstrating example of the present invention, and is not to this
The restriction of bright embodiment.For those of ordinary skill in the field, can also do on the basis of the above description
Go out change or the variation of other multi-form.Here without also cannot all of embodiment be given exhaustive.All in the present invention
Spirit and principle within any amendment, equivalent and the improvement etc. made, should be included in the guarantor of the claims in the present invention
Within the scope of protecting.
Claims (9)
1. based on surface enhanced raman spectroscopy and the glucose sensing approach of bi-molecular probe, it is characterised in that described method bag
Include following steps:
S1: by the coin race metal nano-rod uniform deposition for preparing at carrier surface, is prepared as SERS active-substrate, regulates coin race
The size of metal nano-rod and transverse and longitudinal ratio, make the phasmon wavelength of SERS active-substrate resonate with incident illumination；
S2: by primary glucoreceptor molecular modification SERS active-substrate so that it is be fixed in SERS active-substrate；
SERS active-substrate is soaked in the Glucose standards aqueous solution of variable concentrations, the glucose in solution is captured
In SERS active-substrate；
Adding two grades of glucoreceptor molecules in SERS active-substrate, two grades of glucoreceptor molecules are selective and in substrate
Glucose molecule covalent bond；
Carry out SERS spectra test, demarcate the standard of concentration known with the Raman signatures peak intensity of two grades of glucoreceptor molecules
D/W, sets up quantitative analysis standard curve；
After S3: gather the sample to be tested that concentration of glucose is unknown, suitably dilution, carry out processing and SERS light by the method for step S2
Spectrum test, substitutes into, by the Raman signatures peak intensity of obtain two grades of Fructus Vitis viniferae acceptor molecules, the quantitative analysis standard that step S2 is set up
Curve, obtains the glucose concentration value of sample to be tested.
The most according to claim 1 based on surface enhanced raman spectroscopy with the glucose sensing approach of bi-molecular probe, institute
Shu Bi race metal nano-rod is Jin-silver core-shell nanometer rod.
The most according to claim 1 based on surface enhanced raman spectroscopy with the glucose sensing approach of bi-molecular probe, its
Being characterised by, described carrier is coverslip.
The most according to claim 1 based on surface enhanced raman spectroscopy with the glucose sensing approach of bi-molecular probe, its
Being characterised by, described carrier is filter paper or filter membrane.
The most according to claim 1 based on surface enhanced raman spectroscopy with the glucose sensing approach of bi-molecular probe, its
Being characterised by, the instrument that SERS spectra test uses is Raman spectrometer, and the wave-length coverage of its exciting light is 400 ~ 1000 nm.
The most according to claim 1 based on surface enhanced raman spectroscopy with the glucose sensing approach of bi-molecular probe, its
Being characterised by, described sample to be tested includes blood, urine, saliva, perspiration.
The most according to claim 1 based on surface enhanced raman spectroscopy with the glucose sensing approach of bi-molecular probe, its
Being characterised by, described primary glucoreceptor molecule uses 4-mercaptophenyl boronic acid molecule, and it is fixed in SERS active-substrate.
The most according to claim 1 based on surface enhanced raman spectroscopy with the glucose sensing approach of bi-molecular probe, its
Being characterised by, described two grades of glucoreceptor molecules use 4-cyanophenylboronic acid, itself and the Fructus Vitis viniferae sugar in SERS active-substrate
Son combines.
The most according to claim 8 based on surface enhanced raman spectroscopy with the glucose sensing approach of bi-molecular probe, its
It is characterised by, uses the cyano group raman characteristic peak of 4-cyanophenylboronic acid that concentration of glucose is carried out detection by quantitative.
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Cited By (9)
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|---|---|---|---|---|
| CN106501234A (en) * | 2016-12-20 | 2017-03-15 | 复旦大学 | Cell Raman quiet zone surface enhanced Raman scattering substrate and its preparation method and application |
| CN108680555A (en) * | 2018-04-02 | 2018-10-19 | 上海应用技术大学 | The field fast detection method of metrifonate and Simanex in a kind of seawater |
| CN109001176A (en) * | 2018-06-14 | 2018-12-14 | 福建师范大学 | A kind of preparation method of the SERS substrate of Au@Ag nanoparticle and method using substrate detection glucose |
| WO2021046741A1 (en) * | 2019-09-11 | 2021-03-18 | 亳州市新健康科技有限公司 | Substrate material for high-efficiency surface-enhanced raman scattering and preparation method |
| CN113563222A (en) * | 2021-07-13 | 2021-10-29 | 暨南大学 | Method for detecting hydrogen peroxide based on surface enhanced Raman scattering of silent zone reporter molecule and application |
| CN113892946A (en) * | 2021-09-17 | 2022-01-07 | 天津工业大学 | Preparation method of sweat sensing bandage based on SERS |
| CN114216851A (en) * | 2020-11-27 | 2022-03-22 | 四川大学华西医院 | Acute pancreatitis assessment device based on surface enhanced Raman spectroscopy |
| CN115356323A (en) * | 2022-08-15 | 2022-11-18 | 滨州医学院 | Method for growing gold nanorod array by evaporation-induced self-assembly and SERS (surface enhanced Raman Scattering) sensor for detecting blood sugar or glucose |
| JP7359227B2 (en) | 2020-01-27 | 2023-10-11 | 日本電気株式会社 | A method for sensing plant hormones and a method for early detection of plant disease infection using the same |
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Cited By (13)
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| CN106501234A (en) * | 2016-12-20 | 2017-03-15 | 复旦大学 | Cell Raman quiet zone surface enhanced Raman scattering substrate and its preparation method and application |
| CN108680555A (en) * | 2018-04-02 | 2018-10-19 | 上海应用技术大学 | The field fast detection method of metrifonate and Simanex in a kind of seawater |
| CN108680555B (en) * | 2018-04-02 | 2021-07-20 | 上海应用技术大学 | On-site rapid detection method for trichlorfon and simazine in seawater |
| CN109001176A (en) * | 2018-06-14 | 2018-12-14 | 福建师范大学 | A kind of preparation method of the SERS substrate of Au@Ag nanoparticle and method using substrate detection glucose |
| CN109001176B (en) * | 2018-06-14 | 2021-08-24 | 福建师范大学 | Preparation method of SERS substrate of Au @ Ag nanoparticles and method for detecting glucose by using substrate |
| WO2021046741A1 (en) * | 2019-09-11 | 2021-03-18 | 亳州市新健康科技有限公司 | Substrate material for high-efficiency surface-enhanced raman scattering and preparation method |
| JP7359227B2 (en) | 2020-01-27 | 2023-10-11 | 日本電気株式会社 | A method for sensing plant hormones and a method for early detection of plant disease infection using the same |
| CN114216851A (en) * | 2020-11-27 | 2022-03-22 | 四川大学华西医院 | Acute pancreatitis assessment device based on surface enhanced Raman spectroscopy |
| CN113563222A (en) * | 2021-07-13 | 2021-10-29 | 暨南大学 | Method for detecting hydrogen peroxide based on surface enhanced Raman scattering of silent zone reporter molecule and application |
| CN113563222B (en) * | 2021-07-13 | 2023-05-30 | 暨南大学 | Method for detecting hydrogen peroxide by surface enhanced Raman scattering based on silent region reporter molecule and application |
| CN113892946A (en) * | 2021-09-17 | 2022-01-07 | 天津工业大学 | Preparation method of sweat sensing bandage based on SERS |
| CN113892946B (en) * | 2021-09-17 | 2023-10-31 | 天津工业大学 | Preparation method of sweat sensing binding band based on SERS |
| CN115356323A (en) * | 2022-08-15 | 2022-11-18 | 滨州医学院 | Method for growing gold nanorod array by evaporation-induced self-assembly and SERS (surface enhanced Raman Scattering) sensor for detecting blood sugar or glucose |
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Application publication date: 20161116 |