CN103869021A - High performance liquid chromatography detection method and device based on surface enhanced Raman spectroscopy - Google Patents
High performance liquid chromatography detection method and device based on surface enhanced Raman spectroscopy Download PDFInfo
- Publication number
- CN103869021A CN103869021A CN201410121804.7A CN201410121804A CN103869021A CN 103869021 A CN103869021 A CN 103869021A CN 201410121804 A CN201410121804 A CN 201410121804A CN 103869021 A CN103869021 A CN 103869021A
- Authority
- CN
- China
- Prior art keywords
- high performance
- performance liquid
- liquid chromatography
- raman spectroscopy
- surface enhanced
- 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.)
- Granted
Links
- 238000004128 high performance liquid chromatography Methods 0.000 title claims abstract description 45
- 238000004416 surface enhanced Raman spectroscopy Methods 0.000 title claims abstract description 28
- 238000001514 detection method Methods 0.000 title claims abstract description 27
- 239000011521 glass Substances 0.000 claims abstract description 47
- 238000001069 Raman spectroscopy Methods 0.000 claims abstract description 20
- 150000001875 compounds Chemical class 0.000 claims abstract description 14
- 239000007788 liquid Substances 0.000 claims abstract description 13
- 239000010931 gold Substances 0.000 claims abstract description 8
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims abstract description 4
- 229910052737 gold Inorganic materials 0.000 claims abstract description 4
- 239000002245 particle Substances 0.000 claims description 31
- 239000000243 solution Substances 0.000 claims description 23
- 238000000034 method Methods 0.000 claims description 12
- 238000010521 absorption reaction Methods 0.000 claims description 11
- 239000012071 phase Substances 0.000 claims description 10
- SJECZPVISLOESU-UHFFFAOYSA-N 3-trimethoxysilylpropan-1-amine Chemical compound CO[Si](OC)(OC)CCCN SJECZPVISLOESU-UHFFFAOYSA-N 0.000 claims description 9
- 239000007791 liquid phase Substances 0.000 claims description 9
- 239000011259 mixed solution Substances 0.000 claims description 9
- 238000002360 preparation method Methods 0.000 claims description 7
- 238000001228 spectrum Methods 0.000 claims description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 7
- 238000004140 cleaning Methods 0.000 claims description 6
- 230000004913 activation Effects 0.000 claims description 5
- 238000010168 coupling process Methods 0.000 claims description 4
- 238000005406 washing Methods 0.000 claims description 4
- 230000008878 coupling Effects 0.000 claims description 3
- 238000005859 coupling reaction Methods 0.000 claims description 3
- 239000003480 eluent Substances 0.000 claims description 3
- 238000012545 processing Methods 0.000 claims description 3
- 238000007747 plating Methods 0.000 claims description 2
- 239000002105 nanoparticle Substances 0.000 abstract description 12
- 239000000126 substance Substances 0.000 abstract description 6
- 238000001338 self-assembly Methods 0.000 abstract description 4
- 238000005259 measurement Methods 0.000 abstract description 2
- 230000007547 defect Effects 0.000 abstract 1
- RMVRSNDYEFQCLF-UHFFFAOYSA-N thiophenol Chemical compound SC1=CC=CC=C1 RMVRSNDYEFQCLF-UHFFFAOYSA-N 0.000 description 21
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 12
- 239000000758 substrate Substances 0.000 description 10
- 239000000523 sample Substances 0.000 description 9
- 238000004611 spectroscopical analysis Methods 0.000 description 8
- 238000011160 research Methods 0.000 description 6
- 229910021642 ultra pure water Inorganic materials 0.000 description 6
- 239000012498 ultrapure water Substances 0.000 description 6
- ZVAVRBUTFWLEBK-UHFFFAOYSA-N NC1=CC(=C(C=C1)S)C1=CC=CC=C1 Chemical compound NC1=CC(=C(C=C1)S)C1=CC=CC=C1 ZVAVRBUTFWLEBK-UHFFFAOYSA-N 0.000 description 5
- 230000014759 maintenance of location Effects 0.000 description 5
- 238000000926 separation method Methods 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 244000061458 Solanum melongena Species 0.000 description 4
- 230000002708 enhancing effect Effects 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 4
- 239000001509 sodium citrate Substances 0.000 description 4
- 238000003756 stirring Methods 0.000 description 4
- 230000008859 change Effects 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000004811 liquid chromatography Methods 0.000 description 3
- 238000005728 strengthening Methods 0.000 description 3
- SJUCACGNNJFHLB-UHFFFAOYSA-N O=C1N[ClH](=O)NC2=C1NC(=O)N2 Chemical compound O=C1N[ClH](=O)NC2=C1NC(=O)N2 SJUCACGNNJFHLB-UHFFFAOYSA-N 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000007812 deficiency Effects 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 2
- 229920002521 macromolecule Polymers 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 230000003595 spectral effect Effects 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 238000001237 Raman spectrum Methods 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 238000004817 gas chromatography Methods 0.000 description 1
- 238000002309 gasification Methods 0.000 description 1
- 238000002173 high-resolution transmission electron microscopy Methods 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 238000011005 laboratory method Methods 0.000 description 1
- 150000002605 large molecules Chemical class 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 238000003822 preparative gas chromatography Methods 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000001845 vibrational spectrum Methods 0.000 description 1
Images
Landscapes
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
The invention discloses a high performance liquid chromatography detection method and device based on a surface enhanced Raman spectroscopy. A sample outflowing end opening of a high performance liquid chromatograph is communicated with the inlet end of a detection pipe through a pipeline, the outlet end of the detection pipe is connected with a sample inlet end of a Raman spectroscopy, and the detection pipe is a gold nano particle modified glass pipe, and is applied to liquid chromatogram-surface enhanced Raman spectroscopy detection. When a sample to be detected is detected, by adopting the detection pipe which is prepared by nano particles through self assembly as a high performance liquid chromatography detector device, a multi-component substance can be effectively separated through the high performance liquid chromatography, the defect that the high performance liquid chromatography can not be used for identifying a molecular structure can be overcome, and separated compounds are subjected to surface enhanced Raman spectroscopy discrimination and concentration measurement; the invention provides a novel detection device suitable for the high performance liquid chromatography.
Description
Technical field
The invention belongs to the field of on-line analysis detection technique, be specifically related to a kind of high-efficient liquid phase color spectrum detection device based on Surface enhanced raman spectroscopy.
Background technology
Nineteen sixty, for the macromolecular substances of the difficult gasification such as isolated protein, nucleic acid, the theory of gas chromatography and method are reintroduced back to classical liquid chromatography.1971, the people such as section's Crane (Kirkland) published " modern practice of liquid chromatography " book, indicated that high performance liquid chromatography (HPLC) formally sets up.
Through the development of decades, because high performance liquid chromatography has higher separation efficiency, well selectivity, higher detection sensitivity and wide range of application, become the indispensable analyzing and testing instrument of chemistry and association area.Compared with vapor-phase chromatography, high performance liquid chromatography is volatility and the thermal stability limit of test sample not, applied range; Mobile phase kind is many, can reach high separation efficiency by the optimization of mobile phase; Generally at room temperature analyze, do not need high column temperature, thereby be applicable to analyze the compound of higher boiling, large molecule, strong polarity, poor heat stability.
Although high performance liquid chromatography has above plurality of advantages, but inevitably still come with some shortcomings, as cannot be differentiated the design feature of different compounds, multiple different compounds may have identical retention time etc., cause thus the expansion research of scientists to high performance liquid chromatography detecting device, High Performance Liquid Chromatography/Mass Spectrometry (HPLC-MS) coupling technique of for example generally applying at present, its core is to differentiate means for high performance liquid chromatography provides follow-up compound structure.
Surface enhanced raman spectroscopy (SERS) is as one of the sensitiveest detection technique, and (referring to document: Nature Biotechnology 26,83 – 90 (2008)) receives publicity in various fields.The key that realizes SERS high-sensitivity detection is the surface reinforced Raman active substrate that preparation enhancing ability is strong, stability is good, can obtain sensitive, stable SERS signal on the one hand, expand the range of application of SERS, can also establish on the other hand quantitative measurement according to (referring to document: Physical Review E 62,4318 – 4324 (2000)).And key in strengthening as SERS, the pattern feature of nano particle and arranging has determined the quality of strengthening the property.In the method for common structure nano-device, there is the one nano particle assemble method of " from bottom to top ", the i.e. self assembly of nano particle efficiently, fast and cheaply.Its cardinal principle be by with the effect of difunctional molecule, its one end is closely connected with substrate, and the functional group of the other end can produce chemistry or electrostatic force with nanoparticle surface, thereby form one deck nano particle coverlay at substrate surface.By two dimension or the three-dimensional package assembly of nano particle, can obtain the even and stable SERS substrate on definite meaning.
Therefore, need research to strengthen the preparation of substrate by special surface, a kind of New-type detector that can realize Surface enhanced raman spectroscopy as high performance liquid chromatography is provided, thereby under simple experiment condition, complete the recognition function to molecule in high efficiency liquid phase efflux, there is range of application widely to meet high performance liquid chromatography.
Summary of the invention
Problem to be solved by this invention is to overcome existing high performance liquid chromatography to detect the deficiency existing aspect compound structure discriminating, a kind of eigen vibration signal that can obtain some contained molecule is provided, reaches the high-efficient liquid phase color spectrum detection device based on Surface enhanced raman spectroscopy of differentiating molecular structure function.
To achieve the above object of the invention, the technical solution used in the present invention is to provide a kind of high-efficient liquid phase color spectrum detection device based on Surface enhanced raman spectroscopy, and it comprises high performance liquid chromatograph, Raman spectrometer; Sample outflow port at high performance liquid chromatograph is connected with the entrance point of detector tube by pipeline, and the endpiece of detector tube is connected with the sample inlet end of Raman spectrometer by pipeline; Described detector tube is the glass tube that golden nanometer particle is modified, and preparation method is: the glass tube after washing is placed in to H
20, NH
4oH and H
2o
2mixed solution in, by volume, H
20:NH
4oH:H
2o
2for 5:1:1, be activation processing 0.5～1 hour under the water bath condition of 60～80 ℃ in temperature, obtain the glass tube of coupling aminopropyl trimethoxysilane molecule; Glass tube is placed in to aurosol solution again and soaks 10～12 hours, golden nanometer particle is fixed on glass tube inwall, obtains Surface enhanced raman spectroscopy detector tube through cleaning; The mean diameter of described aurosol GOLD FROM PLATING SOLUTION particle is 45～65nm.
The internal diameter that a preferred version of the present invention is described glass tube is 0.91～1mm, and wall thickness is 0.1～0.15mm, and pipe range is 80～120mm.
Technical solution of the present invention also comprises the method detecting with said apparatus, comprises the steps:
1, detected sample enters chromatographic column through the injector of high performance liquid chromatography, through the adjusting of pump, mobile phase is passed through to chromatographic column continuously, and wash-out is adsorbed in the solute molecule in chromatographic column;
2, the eluent obtaining enters the UV-detector that high performance liquid chromatography is equipped with, and detects in real time the uv absorption signal of efflux;
3, the sample to be checked high performance liquid chromatograph outflow port being flowed out enters Raman spectrometer after the detector tube of the fixing golden nanometer particle of inwall, strengthen Raman shift and the intensity of vibration peak in spectrogram by detecting compound to be checked surface, obtain structure and the concentration of compound to be detected.
The present invention adopts H by volume
20:NH
4oH:H
2o
2the mixed solution of=5:1:1 carries out activation processing to detector tube, and detector tube inwall coupling aminopropyl trimethoxysilane (APTMS) molecule makes the exposed a large amount of hydroxyl of inside pipe wall, utilizes the chemical interactions of silicon in hydroxyl and APTMS, fixing one deck APTMS molecule; The fixing acting force of golden nanometer particle is chemical bond power, by effect amino and golden in APTMS, golden nanometer particle is fixed on to glass detector tube inwall, surface reinforced Raman active substrate detector tube is connected with high performance liquid chromatograph, and the Raman spectrometer that is coupled, can realize chromatographic resolution and spectral detection and carry out in real time.
Principle of the present invention is: utilize the detector tube that is assembled with golden nanometer particle, can be by the efflux after separating in high performance liquid chromatography by this detector tube, efflux by the different retention times of valving control flows through the detector tube that is assembled with golden nanometer particle separately, if the substance separating is adsorbed on golden nanometer particle surface and reaches certain concentration, just can obtain some chemical bond vibrational spectra peak of this molecule by Surface enhanced raman spectroscopy, thereby the structure of compound is differentiated and played a role; Read according to the variation of spectral intensity the concentration that flows out species simultaneously.The surface reinforced Raman active substrate that the present invention is prepared by self-assembly is as a kind of high performance liquid chromatography detecting device, solve high performance liquid chromatography and cannot differentiate the deficiency of molecular structure, some eigen vibration pattern of institute's separating compound is realized to identification, thereby make that high performance liquid chromatography and these two kinds of technology of Surface enhanced raman spectroscopy complement each other, effectively combination.
Due to the employing of technique scheme, compared with prior art, tool of the present invention has the following advantages:
1, the method that adopts Chemical self-assembly, is assembled into glass detector tube inwall by golden nanometer particle, obtains enhancing substrate in a big way, and can realize the effect of online detection.
2, the present invention is by high performance liquid chromatography and the effective combination of Surface enhanced raman spectroscopy, first adopt high performance liquid chromatography to separate potpourri, recycling Surface enhanced raman spectroscopy is identified the eigen vibration of compound in liquid phase stream fluid, the mensuration of implementation structure and concentration, be compared to UV-detector simultaneously, use Surface enhanced raman spectroscopy can realize the detection of the different molecular with identical retention time, and the detectability of Surface enhanced raman spectroscopy is lower than other detection techniques, thereby the analysis of some low concentration compound is played to actual meaning.
3, pick-up unit provided by the invention can utilize the existing checkout equipment in laboratory, and detection method is simple, mild condition, is conducive to apply.
Accompanying drawing explanation
The structural representation of the pick-up unit that Fig. 1 provides for the embodiment of the present invention;
The mean grain size that the sodium citrate reducing process that Fig. 2 provides for the embodiment of the present invention prepares is about the TEM figure of the golden nanometer particle of 55 nm;
Fig. 3 assembles golden nanometer particle front and back glass detector tube variation diagram in kind in the embodiment of the present invention; Wherein, figure a is the glass tube before assembling Au nano particle, and figure b is the glass detector tube after assembling Au nano particle;
Fig. 4 is the prepared SEM figure that is assembled with golden nanometer particle glass detector tube of the embodiment of the present invention;
Fig. 5 be the embodiment of the present invention prepared be assembled with the surface increasing Raman spectrum figure of golden nanometer particle glass detector tube for different probe molecule; Wherein, a is to sulfydryl benzenethiol, and b is 4,4-dipyridine;
Fig. 6 is that the embodiment of the present invention adopts high performance liquid chromatography to separate 4,4-dipyridine and the uv absorption result collection of illustrative plates to sulfydryl benzenethiol;
Fig. 7 is that the glass detector tube that is assembled with golden nanometer particle prepared by the embodiment of the present invention applies to high performance liquid chromatography separation 4,4-dipyridine and the SERS of sulfydryl benzenethiol efflux is detected to collection of illustrative plates;
Fig. 8 is that the embodiment of the present invention adopts high performance liquid chromatography to separate the uv absorption result collection of illustrative plates of 4,4-dipyridine and p-aminophenyl thiophenol;
Fig. 9 is the SERS detection collection of illustrative plates that the prepared glass detector tube that is assembled with golden nanometer particle of the embodiment of the present invention applies to high performance liquid chromatography separation 4,4-dipyridine and p-aminophenyl thiophenol efflux.
Embodiment
Below in conjunction with drawings and Examples, the invention will be further described.
Embodiment 1
Referring to accompanying drawing 1, it is the structural representation of a kind of high-efficient liquid phase color spectrum detection device based on Surface enhanced raman spectroscopy provided by the invention.The sample introduction solution of high performance liquid chromatography enters chromatographic column via the injector of high performance liquid chromatography, afterwards, the adjusting of mobile phase by pump continuously by chromatographic column and make to be adsorbed in solute molecule in chromatographic column along with flowing of mobile phase by wash-out, afterwards, eluent enters the UV-detector that high performance liquid chromatography is equipped with, detect in real time the uv absorption signal of efflux, enter via polytetrafluoro connecting pipe the glass detector tube being positioned in Raman instrument detection platform again, detect in real time the surface-enhanced Raman signal of solute molecule.
Utilize above-mentioned detection device to detect, its step is as follows:
(1) in 100 ml three-neck flasks, adding 100 ml concentration is the chlorauric acid solution of 25 mM, and it is faint yellow that solution is; This solution is heated to boil, and under vigorous stirring, to add 0.75ml concentration be the sodium citrate solution of 0.01 g/ml.Now solution is gradually by the faint yellow black that changes into, then becomes the slightly aubergine of the colour of loess from black, and whole change color process is 1～3 min.Become aubergine with solution and start timing, under stirring condition, keep system fluidized state 15 min, naturally cool to subsequently room temperature.Referring to accompanying drawing 2, it is the TEM figure that the present embodiment adopts golden nanometer particle prepared by sodium citrate reducing process, adopts the Dutch TEcnaiG20 of FEI Co. type high resolution transmission electron microscopy, as seen from Figure 2, it is spherical that golden nanometer particle is, and particle diameter is even, and monodispersity is good.
(2) get simple glass pipe, after within ultrapure water ultrasonic ten minutes, cleaning, use H
20:NH
4oH:H
2o
2=5:1:1 mixed solution, 60～80 ℃ of water-baths activation 0.5～1 hour, is immersed in after washing in APTMS solution and keeps spending the night, after cleaning by 120 ℃ of oven 0.5～1 hour.
(3) get step (1) in the 55nm aurosol centrifugal concentrating of preparation, remove supernatant liquor, step is full of to the aurosol after concentrated in (2) in prepared glass tube, keep approximately 10～12 hours.
(4) the liquid sucking-off in glass tube step being prepared in (3), ultrapure water cleans glass tube 2～3 times, just obtained inwall and be assembled with the glass tube substrate of Au nano particle.The glass tube preparing is kept in ultrapure water.Referring to accompanying drawing 3, it is before and after glass tube assembling golden nanometer particle, to change pictorial diagram in this step, and wherein, a figure is the glass tube before assembling, is water white transparency, and b figure is the glass tube after assembling, is claret, as detector tube.
(5) utilize high performance liquid chromatography to separate 4,4-dipyridine and to sulfydryl benzenethiol mixed solution, mobile phase adopts 50% acetonitrile and 50% water, flow velocity 1mL/min.
The high-efficient liquid phase color spectrum detection device character based on Surface enhanced raman spectroscopy to above-mentioned preparation characterizes, and result is as follows:
Referring to accompanying drawing 4, it is the SEM figure of the prepared glass detector tube that is assembled with golden nanometer particle of the present embodiment; The Hitachi S-4700N scanning electron microscope that adopts Japanese Hitachi company to produce, as shown in Figure 4, glass detector tube inwall gold nanometer particle grain size approximately 55 nm, favorable dispersibility, there is multi-Layer Phenomenon in subregion.
Referring to accompanying drawing 5, it is the research of strengthening the property of the surface of the prepared glass detector tube that is assembled with golden nanometer particle of the present embodiment; Adopt the HR800 type Raman spectrometer of French Jobin Yvon company; The enhancing signal of test different probe molecule.As shown in Figure 5, enhancing signal is good.Wherein, a figure is 10
-3mol/L to sulfydryl benzenethiol, b figure is 10
-34 of mol/L, 4-dipyridine.
Referring to accompanying drawing 6, it is in the present embodiment 4,4-dipyridine and the high performance liquid chromatography Separation Research to sulfydryl benzenethiol mixed solution; Adopt 1200 type high performance liquid chromatographs of Agilent company of the U.S.; Detecting device adopts UV-detector, and mobile phase is 50% acetonitrile and 50% water, separates 4 of same concentrations, 4-dipyridine and to sulfydryl benzenethiol solution, as shown in Figure 6, the corresponding two kinds of different materials in the absorption peak position of uv absorption spectrogram.
Referring to accompanying drawing 7, it is the Surface enhanced raman spectroscopy research of the prepared glass detector tube that is assembled with golden nanometer particle of the present embodiment for high performance liquid chromatography efflux; Adopt the HR800 type Raman spectrometer of French Jobin Yvon company; As shown in Figure 7, in the retention time corresponding to ultraviolet absorption peak, the surface enhanced Raman spectroscopic signals of corresponding molecule can be detected.
The present embodiment carries out detecting based on the high performance liquid chromatography of Surface enhanced raman spectroscopy to sample, and as described in Example 1, detecting step is as follows for pick-up unit:
(1) in 100 ml three-neck flasks, adding 100 ml concentration is the chlorauric acid solution of 25 mM, and it is faint yellow that solution is; This solution is heated to boil, and under vigorous stirring, to add 0.75ml concentration be the sodium citrate solution of 0.01 g/ml.Now solution is gradually by the faint yellow black that changes into, then becomes the slightly aubergine of the colour of loess from black, and whole change color process is 1～3 min.Become aubergine with solution and start timing, under stirring condition, keep system fluidized state 15 min, naturally cool to subsequently room temperature.
(2) get simple glass pipe, after within ultrapure water ultrasonic ten minutes, cleaning, use H
20:NH
4oH:H
2o
2=5:1:1 mixed solution, 60～80 ℃ of water-baths activation 0.5～1 hour, is immersed in after washing in APTMS solution and keeps spending the night, after cleaning by 120 ℃ of oven 0.5～1 hour.
(3) get step (1) in the 55nm aurosol centrifugal concentrating of preparation, remove supernatant liquor, utilize capillarity that step is full of to the aurosol after concentrated in (2) in prepared glass tube, keep approximately 10～12 hours.
(4) the liquid sucking-off in glass tube step being prepared in (3), ultrapure water cleans glass tube 2～3 times, just obtained inwall and be assembled with the glass detector tube substrate of Au nano particle.The glass detector tube preparing is kept in ultrapure water.
(5) utilize high performance liquid chromatography to separate 4,4-dipyridine and p-aminophenyl thiophenol mixed solution, mobile phase adopts 30% acetonitrile and 70% water, flow velocity 1mL/min.
Referring to accompanying drawing 8, it is in the present embodiment 4, the high performance liquid chromatography separating resulting of 4-dipyridine and p-aminophenyl thiophenol mixed solution; Adopt 1200 type high performance liquid chromatographs of Agilent company of the U.S.; Detecting device adopts UV-detector, and mobile phase is 30% acetonitrile and 70% water, separates 4 of same concentrations, 4-dipyridine and p-aminophenyl thiophenol solution, as shown in Figure 8, the corresponding two kinds of different materials in the absorption peak position of uv absorption spectrogram.
Referring to accompanying drawing 9, it is the Surface enhanced raman spectroscopy research of the prepared glass detector tube that is assembled with golden nanometer particle of this example for high performance liquid chromatography efflux; Adopt the HR800 type Raman spectrometer of French Jobin Yvon company; As shown in Figure 9, in the retention time corresponding to ultraviolet absorption peak, the surface enhanced Raman spectroscopic signals of corresponding molecule can be detected.
Claims (3)
1. the high-efficient liquid phase color spectrum detection device based on Surface enhanced raman spectroscopy, it comprises high performance liquid chromatograph, Raman spectrometer, it is characterized in that: the sample outflow port at high performance liquid chromatograph is connected with the entrance point of detector tube by pipeline, the endpiece of detector tube is connected with the sample inlet end of Raman spectrometer by pipeline; Described detector tube is the glass tube that golden nanometer particle is modified, and preparation method is: the glass tube after washing is placed in to H
20, NH
4oH and H
2o
2mixed solution in, by volume, H
20:NH
4oH:H
2o
2for 5:1:1, be activation processing 0.5～1 hour under the water bath condition of 60～80 ℃ in temperature, obtain the glass tube of coupling aminopropyl trimethoxysilane molecule; Glass tube is placed in to aurosol solution again and soaks 10～12 hours, golden nanometer particle is fixed on glass tube inwall, obtains Surface enhanced raman spectroscopy detector tube through cleaning; The mean diameter of described aurosol GOLD FROM PLATING SOLUTION particle is 45～65nm.
2. a kind of high performance liquid chromatography detecting device based on Surface enhanced raman spectroscopy according to claim 1, is characterized in that: the internal diameter of described glass tube is 0.91～1mm, and wall thickness is 0.1～0.15mm, and pipe range is 80～120mm.
3. the method detecting with the high-efficient liquid phase color spectrum detection device based on Surface enhanced raman spectroscopy claimed in claim 1, is characterized in that comprising the steps:
(1) detected sample enters chromatographic column through the injector of high performance liquid chromatography, through the adjusting of pump, mobile phase is passed through to chromatographic column continuously, and wash-out is adsorbed in the solute molecule in chromatographic column;
(2) eluent obtaining enters the UV-detector that high performance liquid chromatography is equipped with, and detects in real time the uv absorption signal of efflux;
(3) sample to be checked high performance liquid chromatograph outflow port being flowed out enters Raman spectrometer after the detector tube of the fixing golden nanometer particle of inwall, strengthen Raman shift and the intensity of vibration peak in spectrogram by detecting compound to be checked surface, obtain structure and the concentration of compound to be detected.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410121804.7A CN103869021B (en) | 2014-03-28 | 2014-03-28 | Based on high-efficiency liquid chromatography method for detecting and the device of Surface enhanced raman spectroscopy |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410121804.7A CN103869021B (en) | 2014-03-28 | 2014-03-28 | Based on high-efficiency liquid chromatography method for detecting and the device of Surface enhanced raman spectroscopy |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103869021A true CN103869021A (en) | 2014-06-18 |
| CN103869021B CN103869021B (en) | 2015-11-04 |
Family
ID=50907782
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410121804.7A Expired - Fee Related CN103869021B (en) | 2014-03-28 | 2014-03-28 | Based on high-efficiency liquid chromatography method for detecting and the device of Surface enhanced raman spectroscopy |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103869021B (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107655873A (en) * | 2016-07-25 | 2018-02-02 | 岛津分析技术研发（上海）有限公司 | Raman spectrum analysis device and method |
| CN109342602A (en) * | 2018-11-21 | 2019-02-15 | 中国计量大学 | Aflatoxin detection device in a kind of liquid |
| CN114923996A (en) * | 2022-04-27 | 2022-08-19 | 苏州大学 | Automatic HPLC-SERS combined device based on paper substrate and detection method |
| CN116165188A (en) * | 2022-12-12 | 2023-05-26 | 苏州大学 | Automatic HPLC-SERS combined system based on liquid phase substrate and application thereof |
| CN117589687A (en) * | 2024-01-18 | 2024-02-23 | 成都艾立本科技有限公司 | Optical cavity container based on air-wrapped liquid, application and spectrum detection method |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102128908A (en) * | 2010-01-19 | 2011-07-20 | 华东理工大学 | On-site quick detection method for organic pollutants in water |
| CN103398995A (en) * | 2012-09-07 | 2013-11-20 | 欧普图斯(苏州)光学纳米科技有限公司 | Integrated equipment capable of realizing chemical separation and light dispersion |
-
2014
- 2014-03-28 CN CN201410121804.7A patent/CN103869021B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102128908A (en) * | 2010-01-19 | 2011-07-20 | 华东理工大学 | On-site quick detection method for organic pollutants in water |
| CN103398995A (en) * | 2012-09-07 | 2013-11-20 | 欧普图斯(苏州)光学纳米科技有限公司 | Integrated equipment capable of realizing chemical separation and light dispersion |
Non-Patent Citations (7)
| Title |
|---|
| BERND SAGMULLER ET AL.: "Identification of illicit drugs by a combination of liquid chromatography and surface-enhanced Raman scattering spectroscopy", 《JOURNAL OF MOLECULAR STRUCTURE》 * |
| BERNHARD ZACHHUBER ET AL.: "Quantification of DNT isomers by capillary liquid chromatography using at-line SERS detection or multivariate analysis of SERS spectra of DNT isomer mixtures", 《J. RAMAN SPECTROSC.》 * |
| CAROLINA CARRILLO-CARRION ET AL.: "Determination of Pyrimidine and Purine Bases by Reversed-Phase Capillary Liquid Chromatography with At-Line Surface-Enhanced Raman Spectroscopic Detection Employing a Novel SERS Substrate Based on ZnS/CdSe Silver-Quantum Dots", 《ANAL. CHEM.》 * |
| 徐燕慧 等: "干净、均一的表面增强拉曼基底的制备和表征", 《光散射学报》 * |
| 盛蓉生 等: "核酸碱基和核普的高效液相色——谱表面增强拉曼光谱在线联机分析研究", 《光散射学报》 * |
| 高敏侠 等: "结合化学组装和电沉积的SERS基底的制备方法", 《高等学校化学学报》 * |
| 龚龑 等: "拉曼光谱技术在色谱分析检测中的应用", 《化学通报》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107655873A (en) * | 2016-07-25 | 2018-02-02 | 岛津分析技术研发（上海）有限公司 | Raman spectrum analysis device and method |
| CN109342602A (en) * | 2018-11-21 | 2019-02-15 | 中国计量大学 | Aflatoxin detection device in a kind of liquid |
| CN114923996A (en) * | 2022-04-27 | 2022-08-19 | 苏州大学 | Automatic HPLC-SERS combined device based on paper substrate and detection method |
| CN116165188A (en) * | 2022-12-12 | 2023-05-26 | 苏州大学 | Automatic HPLC-SERS combined system based on liquid phase substrate and application thereof |
| CN116165188B (en) * | 2022-12-12 | 2024-02-09 | 苏州大学 | Automatic HPLC-SERS combined system based on liquid phase substrate and application thereof |
| CN117589687A (en) * | 2024-01-18 | 2024-02-23 | 成都艾立本科技有限公司 | Optical cavity container based on air-wrapped liquid, application and spectrum detection method |
| CN117589687B (en) * | 2024-01-18 | 2024-04-09 | 成都艾立本科技有限公司 | Optical cavity container based on air-wrapped liquid, application and spectrum detection method |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103869021B (en) | 2015-11-04 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Wang et al. | Chip-based array magnetic solid phase microextraction on-line coupled with inductively coupled plasma mass spectrometry for the determination of trace heavy metals in cells | |
| Majedi et al. | Recent advances in the separation and quantification of metallic nanoparticles and ions in the environment | |
| Dias et al. | Molecularly imprinted polymer as a solid phase extractor in flow analysis | |
| CN103869021B (en) | Based on high-efficiency liquid chromatography method for detecting and the device of Surface enhanced raman spectroscopy | |
| Özkantar et al. | Spectrophotometric detection of rhodamine B in tap water, lipstick, rouge, and nail polish samples after supramolecular solvent microextraction | |
| EP2503335B1 (en) | Method and device for detecting analytes | |
| Alexovič et al. | A single-valve sequential injection manifold (SV-SIA) for automation of air-assisted liquid-phase microextraction: stopped flow spectrophotometric determination of chromium (vi) | |
| CN102262163B (en) | Rapid and automatic determination method and device for tripolycyanamide content in dairy products | |
| CN105466905B (en) | A kind of surface-enhanced Raman detection chip of integrated solid-phase extraction column | |
| Davis et al. | Low-cost and open-source strategies for chemical separations | |
| Müller et al. | Nanoparticle separation with a miniaturized asymmetrical flow field-flow fractionation cartridge | |
| CN106238109A (en) | The micro-fluidic chip of a kind of methamphetamine hydrochloride in Raman detection hair and using method thereof | |
| Pourbasheer et al. | Dispersive liquid–liquid microextraction for preconcentration and determination of phenytoin in real samples using response surface methodology-high performance liquid chromatography | |
| Hartwell et al. | Bead injection with a simple flow-injection system: an economical alternative for trace analysis | |
| CN102565170A (en) | Sensitive capillary electrophoresis methods for detecting melamine in formula milk powder | |
| CN115266627B (en) | Ultraviolet detection method for rapidly measuring mixed micro-plastic and lead | |
| Zhou et al. | Pretreatment, identification and quantification of submicro/nano-plastics in complex environmental matrices | |
| Fung et al. | Determination of trace metals by capillary electrophoresis | |
| CN110044867A (en) | A kind of fruit surface pesticide residue in-situ acquisition detection device and method | |
| CN105842269B (en) | A kind of device for integrating nmr magnet and popping one's head in | |
| Sanjuan-Navarro et al. | In-tube solid phase microextraction coupled to miniaturized liquid chromatography for both, noble metal nanoparticle assessment and sensitive plasmonic assay development | |
| Wang et al. | Long‐chain ionic liquid based mixed hemimicelles and magnetic dispersed solid‐phase extraction for the extraction of fluorescent whitening agents in paper materials | |
| Tsuyama et al. | Diffraction-based label-free photothermal detector for separation analyses in a nanocapillary | |
| Marina et al. | Derivatization in capillary electrophoresis | |
| CN101358946A (en) | Anionic polymer grafting coatings capillary pipe and analytical method for on-line enrichment for protein |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20170810 Address after: 510530, building sixth, building 11, No. 601-610, Kaiyuan Avenue, Science City, Guangzhou hi tech Industrial Development Zone, Guangdong, Co-patentee after: SHANGHAI CLINMETA CO.,LTD. Patentee after: GUANGZHOU CLIN MASS SPECTRUM MEDICAL INSTRUMENT CO.,LTD. Co-patentee after: SHANGHAI AIKESAIMO MEDICAL INSTRUMENT CO.,LTD. Co-patentee after: JIANGSU CLIN CHROMATICNESS MEDICAL INSTRUMENT CO.,LTD. Address before: 511400, No. five, 5 Street, Dalong village, Dalong street, Guangzhou, Guangdong, Panyu District Patentee before: Guangzhou's Intellectual Property Service Co.,Ltd. Effective date of registration: 20170810 Address after: 511400, No. five, 5 Street, Dalong village, Dalong street, Guangzhou, Guangdong, Panyu District Patentee after: Guangzhou's Intellectual Property Service Co.,Ltd. Address before: 215123 Suzhou City, Suzhou Province Industrial Park, No. love road, No. 199 Patentee before: Soochow University |
|
| TR01 | Transfer of patent right | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: High performance liquid chromatography detection method and device based on surface enhanced Raman spectroscopy Effective date of registration: 20200629 Granted publication date: 20151104 Pledgee: Haizhu sub branch of Guangzhou Rural Commercial Bank Co.,Ltd. Pledgor: SHANGHAI CLINMETA Co.,Ltd.|JIANGSU CLIN CHROMATICNESS MEDICAL INSTRUMENT Co.,Ltd.|SHANGHAI AIKESAIMO MEDICAL INSTRUMENT Co.,Ltd.|GUANGZHOU CLIN MASS SPECTRUM MEDICAL INSTRUMENT Co.,Ltd. Registration number: Y2020980003605 |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20151104 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |