CN107064105A - It is a kind of at the same detect aquatic products in sunset yellow and malachite green method - Google Patents
It is a kind of at the same detect aquatic products in sunset yellow and malachite green method Download PDFInfo
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- CN107064105A CN107064105A CN201710228400.1A CN201710228400A CN107064105A CN 107064105 A CN107064105 A CN 107064105A CN 201710228400 A CN201710228400 A CN 201710228400A CN 107064105 A CN107064105 A CN 107064105A
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- malachite green
- sunset yellow
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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
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/40—Concentrating samples
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/40—Concentrating samples
- G01N2001/4038—Concentrating samples electric methods, e.g. electromigration, electrophoresis, ionisation
Abstract
The invention discloses a kind of while detecting the method for sunset yellow and malachite green in aquatic products.This method comprises the following steps：(1) reduced graphene (RGO) nano material is prepared by hydro-thermal method；(2) gold nanorods (GNRs) are synthesized by seeded growth method；(3) RGO, GNRs, SY, MG in adsorbent solution are distinguished on screen printing electrode using electrochemistry concentration method；(4) raman spectral signal of screen printing electrode surface mass is detected, so as to carry out qualitative and quantitative analysis detection to SY, MG in aquatic products.The present invention not only has that analysis is quick, sensitivity is high, amount of samples is few using electrochemistry enrichment SERS detection techniques, has a wide range of application, it is easy to operate and easy to carry the features such as, above all, the present invention has excellent selectivity, and need not mark and isolate and purify, analysis detection can be carried out to sunset yellow in aquatic products and malachite green simultaneously, and test limit is low.
Description
Technical field
The invention belongs to technical field of analysis and detection, specifically, be related to it is a kind of and meanwhile detect in aquatic products sunset yellow and
The method of malachite green.
Background technology
Sunset yellow (SY) is mainly used in the coloring of food and medicine, but consumption is severely restricted.If people it is long-term or
The exceeded food of pigment content such as disposable a large amount of edible sunset yellows, may cause the symptoms such as allergy, diarrhoea, when intake mistake
It when greatly, more than liver load, can in vivo accumulate, certain injury is produced to kidney, liver.Malachite green (MG) is a kind of poisonous
Triphenylmethane chemicals, be both dyestuff, be also antifungal, bactericidal, the medicine of antiparasitic, therefore fish etc. can be used as
Bactericide, but long-term Use out of range can carcinogenic, teratogenesis and mutagenesis.China is in agricultural industry criteria《NY5071-2002 is without public affairs
Evil food fish medicine usage criteria》In malachite green is also classified as forbidden drugses.It would therefore be highly desirable to set up for SY in aquatic products and MG
Field fast detection method.
SERS (SERS) refers to when some molecules are adsorbed to some textured metals (Au, Ag, Cu etc.)
During surface, their Raman scattering intensities can increase by 104~106Times.The characteristics of due to SERS technology rapid sensitives, it is widely used in
In terms of food security, biological detection.Gold nanorods modification reduced graphene can be used as surface enhanced Raman substrate material.In recent years
Come, Chinese patent (A of publication number CN 104977286) carries out analysis detection as SERS substrates using golden nanometer particle to MG,
But analysis detection is but rarely reported while realizing SY and MG.Graphene is a kind of new two-dimentional carbon nanomaterial, its carbon
Hybrid form between atom is sp2 hydridization.Because graphene has higher surface area and stronger electric conductivity, therefore in electricity
Had broad application prospects in terms of, optics, bio-sensing.
Reduced graphene (RGO), sunset yellow (SY) are negatively charged in the aqueous solution, gold nanorods (GNRs), malachite green
(MG) it is positively charged, therefore, the method that can be enriched with using electrochemical in-situ, selective absorption SY, MG, so as to realize aquatic products
Middle SY and MG field quick detection.Electrochemistry enrichment-SERS technologies have that analyze speed is fast, detection sensitivity is high and choosing simultaneously
The features such as selecting property is good, the technology is expected to further apply environment and food analysis field.
The content of the invention
In order to solve the deficiency in above-mentioned prior art, sunset in aquatic products is detected simultaneously the invention provides a kind of
The method of yellow and malachite green.Sunset yellow and peacock in the inventive method energy high selectivity, enzyme rapidly and sensitively analysis aquatic products
Malachite green.
The purpose of the present invention is achieved through the following technical solutions.
It is a kind of at the same detect aquatic products in sunset yellow and malachite green method, comprise the following steps that：
(1) cyclic voltammetry is used, using three-electrode system, using PBS solution as cushioning liquid, reduction graphite is obtained respectively
Alkene RGO, gold nanorods GNRs, sunset yellow SY and malachite green MG oxidation-reduction potential；Further according to oxidation-reduction potential, selection
Suitable SY accumulating potential and MG accumulating potential so that above-mentioned four kinds of materials not oxidized reduction when being enriched with respectively；
(2) electrochemical workstation is utilized, using i-t methods, screen printing electrode is placed in aquatic product extract, using electricity
Chemical enrichment method applies accumulating potential, is enriched with 0.5~10min so that gold nanorods GNRs and malachite green MG are fully adsorbed onto
Electrode surface, reduced graphene RGO and sunset yellow SY are adsorbed onto electrode surface；
(3) detection of Raman signal is carried out to electrode surface using Portable Raman spectrometer, electrode surface sample is obtained
SERS collection of illustrative plates；Again by itself and sunset yellow SY, the SERS spectral contrasts of malachite green MG solids, and then realize sunset in aquatic products
The yellow qualitative and quantitative detections of SY and malachite green MG.
In the present invention, in step (1), reduced graphene RGO is prepared by hydro-thermal method；Gold nanorods GNRs is given birth to by crystal seed
Regular way is synthesized.
In the present invention, in step (1), sunset yellow SY accumulating potential is 0.01~0.25V；Malachite green MG enrichment electricity
Position is -0.25~-0.01V.
In the present invention, in step (3), excitation wavelength is 785nm, and the time of integration is 1~60s.
Compared with prior art, the beneficial effects of the present invention are：
1st, it is combined by electrochemistry concentration method and SERS technologies, sunset yellow in sample and malachite green can be realized simultaneously
Selective enrichment and detection, the method can not only eliminate the interference of other materials, and can improve the inspection of sunset yellow and malachite green
Survey sensitivity；
2nd, the sunset yellow and malachite green molecule by electrochemistry concentration method in screen printing electrode adsorption solution,
Can significant increase analysis detection sensitivity, the method have it is easy to operate, have wide range of applications, rapidly and efficiently and be easy to carry
Feature, and amount of samples is few, meets the demand of trace detection；
3rd, it is combined, is can be achieved to sunset yellow in product and malachite by electrochemical workstation and Portable Raman spectrometer
Green live fast qualitative and quantitative detection, according to linear between sunset yellow and the concentration and Raman signal intensity of malachite green
Relation, test limit (DL) determines zero standard deviation (δ) according to 3 times and the ratio calculation of linearity curve slope (k) is obtained, i.e. DL
=3 δ/k, the test limit that can obtain sunset yellow and malachite green is respectively 5 × 10-7Mol/L and 5 × 10-9mol/L；According to linear
It is 5 × 10 that curve, which lists quantitative detection Cmax,-3Mol/L, this is upper limit of detection.
Brief description of the drawings
Fig. 1 is the flow chart of the detection method of the embodiment of the present invention.
Fig. 2 is sunset yellow, the SERS collection of illustrative plates of malachite green solid in the embodiment of the present invention.
Fig. 3 is the SERS collection of illustrative plates before and after electrochemistry enrichment sunset yellow in the embodiment of the present invention.
Fig. 4 is the SERS collection of illustrative plates of electrochemistry enrichment various concentrations sunset yellow in the embodiment of the present invention, and (five are marked shown in figure
Angle star) be sunset yellow TuPu method peak.
Fig. 5 is the standard concentration and feature peak intensity (1592 ± 2cm of sunset yellow-1) linear relationship schematic diagram.
Fig. 6 is the SERS collection of illustrative plates before and after electrochemistry enrichment malachite green in the embodiment of the present invention.
Fig. 7 is the SERS collection of illustrative plates of electrochemistry enrichment various concentrations malachite green in the embodiment of the present invention, is marked shown in figure
(five-pointed star) is the TuPu method peak of malachite green.
Fig. 8 is the standard concentration and feature peak intensity (1172 ± 2cm of malachite green-1) linear relationship schematic diagram.
Embodiment
Embodiment 1：Electrochemistry enrichment-surface enhanced Raman technique detection sunset yellow and malachite green
(1) redox graphene is prepared
Graphene oxide is prepared using improved Hummers methods, constant volume obtains the flocculation of 0.5mg/mL graphene oxides, taken
5mL graphene oxides in distilling flask, add 5 μ L hydrazine hydrates (35%) as reducing agent, then add 100 μ L ammoniacal liquor (28%),
It is sufficiently stirred for, and the 1h that flows back at 95 DEG C.Room temperature is stood, 10000rpm centrifugation 20min takes subnatant constant volume to obtain
0.5mg/mL redox graphene solution.
(2) gold nanorods are prepared by crystal seed method
A. gold nano seed solution is prepared：Under room temperature condition (25-28 DEG C), 9.75ml 0.1mol/L cetyls are prepared
The trimethylammonium bromide aqueous solution, uniform stirring is added dropwise 0.25ml 0.01mol/L aqueous solution of chloraurate, treats it in solution to transparent
In it is dispersed after, rapidly join the 0.01mol/L sodium borohydride solutions (ice-water bath) of 0.6mL Fresh, solution is by pale yellow
Discoloration is stored at room temperature standby after 2h into brown color, uniform stirring 3min.
B. gold nanorods solution is prepared and purified：Under room temperature condition, 10ml 0.1mol/L cetyl trimethyl is prepared
Bromination aqueous ammonium, adds 0.5mL 0.01mol/L aqueous solution of chloraurate, and 0.1mL 0.01mol/ are added after being well mixed
L silver nitrates, 0.2mL 1mol/L hydrochloric acid, are sufficiently stirred for, and add 80 μ L 0.1mol/L ascorbic acid, and solution is changed into from buff
It is colourless, the gold seeds solution that 12 μ L have been prepared is added, uniform stirring 3 minutes is stored at room temperature 6h.The gold nanorods prepared lead to
8000rpm centrifugation 5min are crossed, washs three times, removes unnecessary cetyl trimethylammonium bromide.Gained gold nanorods concentration is
1.8nM。
(3) oxidation-reduction potential of four kinds of materials is determined
Using electrochemical workstation, electrode used therein is screen printing electrode, and working electrode and auxiliary electrode are carbon electrode, right
Electrode is silver-silver chloride electrode, using cyclic voltammetry (CV), is comprised the following steps that：
A. the oxidation-reduction potential of graphene：Graphene oxide is dropped on the working electrode of screen printing electrode, it is natural
Dry；With 5mL 0.1mol/L PBS solution (potassium chloride containing 0.1mol/L) for electrolyte, scanning voltage is 0.4~-1.2V,
Sweep speed is 50mv/s, obtains the oxidation-reduction potential of graphene.
B. golden oxidation-reduction potential：Gold is coated in the working electrode surface of screen printing electrode, dried naturally；With 5mL
0.1mol/L PBS solution (potassium chloride containing 0.1mol/L) is electrolyte, and scanning voltage is -0.1~1.2V, and sweep speed is
50mv/s, obtains the oxidation-reduction potential of gold.
C. the oxidation-reduction potential of sunset yellow：5mL 1×10-4Mol/L sunset yellow and PBS mixed solutions is electrolyte,
Scanning voltage is -1~0.2V, and sweep speed is 50mv/s, obtains the oxidation-reduction potential of sunset yellow.
D. the oxidation-reduction potential of malachite green：5mL 1×10-4Mol/L malachite green and PBS mixed solutions is electrolysis
Liquid, scanning voltage is 0.05~0.9V, and sweep speed is 50mV/s, obtains the oxidation-reduction potential of malachite green.
The oxidation-reduction potential of four kinds of materials more than, the optimal accumulating potential of selection sunset yellow is 0.2V：Malachite
Green optimal accumulating potential is -0.2V.
(4) electrochemical method enrichment-SERS detects sunset yellow
Using electrochemical workstation, using i-t methods, screen printing electrode is placed in reaction solution, the reaction solution contains
3mL5×10-4M sunset yellows solution (pH=3~4), 50 μ L redox graphenes (0.5mg/mL), 100 μ L sodium chloride solutions
(0.1mol/L), it is 0.2V to control accumulating potential, and enrichment time is 5min.
To the screen printing electrode after above-mentioned enrichment, the detection of Raman signal is carried out using Portable Raman spectrometer, is swashed
Hair wavelength is 785nm, and the time of integration is 10s, obtains the SERS collection of illustrative plates of sample, the Raman spectral contrast (figure with SY, MG solid
2), so as to realize the detection of sunset yellow.As shown in figure 3, the signal of sunset yellow is remarkably reinforced before and after electrochemistry enrichment.
The sunset yellow standard liquid of various concentrations is prepared ,-SERS is enriched with using electrochemistry and detects its spectral signal, using drawing
Graceful 1393 ± 2cm of spectrum peak-1、1592±2cm-1It is used as the characteristic peak for judging sunset yellow.With in solution to be measured sunset yellow it is dense
Degree is gradually increased (5.0 × 10-7Mol/L~5.0 × 10-3Mol/L), 1393 ± 2cm in Raman spectrogram-1、1592±2cm-1Two
The feature peak intensity at place gradually increases (Fig. 4) therewith, selects with 1592 ± 2cm-1Correspondence peak intensity combination linearity curve (Fig. 5) can
Content to sunset yellow is calculated.
(5) electrochemical method enrichment-SERS detects malachite green
Using electrochemical workstation, using i-t methods, screen printing electrode is placed in reaction solution, the reaction solution contains
3mL5×10-5M malachite green solutions, 100 μ L gold nanorods (1.8nmol/L), 100 μ L sodium chloride solutions (0.1mol/L), control
Accumulating potential processed is -0.2V, and enrichment time is 5min.
To the screen printing electrode after above-mentioned enrichment, the detection of Raman signal is carried out using Portable Raman spectrometer, is swashed
Hair wavelength is 785nm, and the time of integration is 10s, the SERS collection of illustrative plates of acquisition sample, the Raman spectral contrast (Fig. 2) with MG solids, from
And realize the detection of malachite green.As shown in fig. 6, the signal of malachite green is remarkably reinforced before and after electrochemistry enrichment.
The malachite green standard liquid of various concentrations is prepared ,-SERS is enriched with using electrochemistry and detects its spectral signal, is used
1172 ± 2cm of Raman spectral peaks-1、1618±2cm-1It is used as the characteristic peak for judging malachite green.With solution Malachite stone to be measured
Green concentration is gradually increased (5.0 × 10-9Mol/L~5.0 × 10-3Mol/L), 1172 ± 2cm in Raman spectrogram-1、1618±
2cm-1Feature peak intensity at two gradually increases (Fig. 7) therewith, selects with 1172 ± 2cm-1Correspondence peak intensity combination linearity curve
(Fig. 8) can be calculated the content of malachite green.
Embodiment 2：Detect the sunset yellow and malachite green in aquatic products
Fig. 1 schematically illustrates the flow chart of sunset yellow and malachite green in the aquatic products of the embodiment of the present invention, described
Detection method comprises the following steps：
(1) pretreatment of sample is oppressed
Oppress Malachite Green and its extraction of metabolin and comprising the following steps that for purification：Weigh 2.00 ± 0.02g fishes
Meat adds 500 μ L 9.5g/L hydroxylamine hydrochloride solution avoid light place 10min in 50m L centrifuge tubes, and then vortex oscillation makes
Meat is uniformly dispersed.The 1min that is vortexed energetically after 10m L acetonitriles and 1.0 ± 0.1g anhydrous magnesium sulfates is added, is added in 4.0 ± 0.1g
It is put into after property aluminum oxide in shaking table and 10min is vibrated under the conditions of 250rpm.Sample 3220rpm centrifuges 5min afterwards, uses liquid-transfering gun
Supernatant liquor is pipetted into 15m L centrifuge tubes.The extract solution being collected into is dried up under 50 DEG C of water-bath with nitrogen stream, Ran Houyong
1.0mL 0.003mol/L 2,3-Dichloro-5,6-dicyano-1,4-benzoquinone solution (solvent is acetonitrile) dissolution residual substance, slightly
Vibrate after 10min, be transferred in the 1.5mL centrifuge tubes equipped with 0.5g neutral aluminas, 30~60s of vortex mixed is (depending on different fishes
Depending on, make color close to colourless.Finally, rotating speed is that 20000rpm centrifuges 5min, and supernatant is filtered with 0.45 μm of organic phase filter membrane
Liquid.
(2) reduced graphene, step be the same as Example 1 are prepared
(3) gold nanorods, step be the same as Example 1 are prepared and purified
(4) sunset yellow and malachite green in electrochemistry enrichment-SERS technology for detection flesh of fish sample extracting solution
Using electrochemical workstation, using i-t methods, in the extract solution that screen printing electrode is placed in above-mentioned flesh of fish sample,
The reaction solution contains 50 μ L reduced graphenes (0.5mg/mL), 100 μ L gold nanorods (1.8nmol/L), 100 μ L sodium chloride
(0.1mol/L) solution, it is -0.2V to control accumulating potential, and enrichment time is 5min, accumulating potential 0.2V is then controlled, during enrichment
Between be 5min.
To the screen printing electrode after above-mentioned enrichment, the detection of Raman signal is carried out using Portable Raman spectrometer, is swashed
Hair wavelength is 785nm, and the time of integration is 10s, obtains the SERS collection of illustrative plates of flesh of fish sample extracting solution, the standard curve pair with SY, MG
According to so as to obtain the content of SY and MG in flesh of fish sample, testing result is as shown in table 1, testing result and high performance liquid chromatography
Compare, result error is less than 10% between the two, as a result show that the accuracy in detection of this law is preferable, alternative high performance liquid chromatography
Method is applied to the field assay detection of malachite green and sunset yellow.
Table 1
In embodiment, according to the linear relationship between sunset yellow and the concentration and Raman signal intensity of malachite green, according to 3
Zero standard deviation (δ) is determined again and the ratio calculation of linearity curve slope (k) obtains test limit (DL), i.e. DL=3 δ/k can
The test limit for obtaining sunset yellow and malachite green is respectively 5 × 10-7Mol/L and 5 × 10-9mol/L；Listed according to linearity curve
Quantitatively detection Cmax is 5 × 10-3Mol/L, this is upper limit of detection.
It can be used for the method that sunset yellow and malachite green analyze detection simultaneously in aquatic products present approach provides a kind of,
The method has the features such as instrument is simple, easy to operate, measure is quick and with high accuracy, is expected to as a kind of portable analysis
Technology is applied to the field quick detection of food.
Claims (5)
1. it is a kind of while detecting the method for sunset yellow and malachite green in aquatic products, it is characterised in that to comprise the following steps that：
(1) cyclic voltammetry is used, using three-electrode system, using phosphate PBS solution as cushioning liquid, reduction stone is obtained respectively
Black alkene RGO, gold nanorods GNRs, sunset yellow SY and malachite green MG oxidation-reduction potential；Further according to oxidation-reduction potential, choosing
Select suitable sunset yellow SY accumulating potential and malachite green MG accumulating potential so that above-mentioned four kinds of materials are in silk-screen printing electricity
Not oxidized reduction during the surface enrichment of pole；
(2) electrochemical workstation is utilized, using i-t methods, screen printing electrode is placed in aquatic product extract, using electrochemistry
Concentration method applies positive potential so that gold nanorods GNRs and malachite green MG are adsorbed onto screen printing electrode surface；Apply again negative
Current potential so that reduced graphene RGO and sunset yellow SY are adsorbed onto screen printing electrode surface；
(3) detection of Raman spectrum is carried out to electrode surface adsorbent using Portable Raman spectrometer, electrode surface is obtained
The SERS SERS collection of illustrative plates of material；Again by itself and sunset yellow SY, the Raman spectral contrast of malachite green MG solids,
And then realize the qualitative and quantitative detection of sunset yellow SY and malachite green MG in aquatic products.
2. according to the method described in claim 1, it is characterised in that in step (1), reduced graphene RGO passes through hydro-thermal legal system
It is standby；Gold nanorods GNRs is synthesized by seeded growth method.
3. according to the method described in claim 1, it is characterised in that in step (1), sunset yellow SY accumulating potential be 0.01~
0.25V；Malachite green MG accumulating potential is -0.01~-0.25V.
4. according to the method described in claim 1, it is characterised in that in step (3), excitation wavelength is 785nm, the time of integration is
60s。
5. according to the method described in claim 1, it is characterised in that in step (3), with 1592 ± 2cm-1Corresponding peak intensity knot
Linear curve is quantitatively calculated the content of sunset yellow in sample；With 1172 ± 2cm-1Corresponding peak intensity combines linear bent
Line is calculated the content of sample Malachite Green.
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| CN109030456A (en) * | 2018-08-25 | 2018-12-18 | 复旦大学 | A kind of Surface enhanced Raman spectroscopy detection substrate and its preparation method and application |
| CN110487863A (en) * | 2019-09-02 | 2019-11-22 | 扬州大学 | A method of sunset yellow content is detected using graphitized carbon modified electrode |
| CN110562956A (en) * | 2019-09-04 | 2019-12-13 | 复旦大学 | Preparation method and application of ordered mesoporous carbon nanofiber array material |
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| CN110562956A (en) * | 2019-09-04 | 2019-12-13 | 复旦大学 | Preparation method and application of ordered mesoporous carbon nanofiber array material |
| CN113740388A (en) * | 2021-07-22 | 2021-12-03 | 西安交通大学 | Gas-sensitive sensing material based on in-situ enrichment amplification strategy and preparation method and application thereof |
| CN113740388B (en) * | 2021-07-22 | 2022-10-25 | 西安交通大学 | Gas-sensitive sensing material based on in-situ enrichment amplification strategy and preparation method and application thereof |
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