CN108037110A - The method that surface Raman enhancement technology based on silver nanoparticle colloidal sol substrate quickly detects probenazole pesticide in rape - Google Patents
The method that surface Raman enhancement technology based on silver nanoparticle colloidal sol substrate quickly detects probenazole pesticide in rape Download PDFInfo
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- 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
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Abstract
The invention discloses a kind of method that surface Raman enhancement technology based on silver nanoparticle colloidal sol substrate quickly detects probenazole pesticide in rape, including：Step 1, by the rape, acetonitrile and sodium chloride that are attached with probenazole pesticide after mixing, vortex oscillation, centrifugation are carried out successively, obtains supernatant；Step 2, ferriferrous oxide nano-particle is mixed with supernatant, and carries out vortex oscillation, centrifugation, filtering successively, obtain prepare liquid；Step 3, silver nanoparticle colloidal sol substrate is prepared；Step 4, by silver nanoparticle colloidal sol substrate, prepare liquid and sodium-chloride water solution after mixing, Raman spectrum collection is carried out；Step 5, the residual quantity of probenazole pesticide in rape is calculated according to formula.Probenazole pesticide residue detection method provided by the invention, quick, cheap, easy, accurate detection scheme is provided for Detecting Pesticide.
Description
Technical field
The present invention relates to plant pesticide residue detection technical field, and in particular to a kind of table based on silver nanoparticle colloidal sol substrate
The method that face Raman enhancing technology quickly detects probenazole pesticide in rape.
Background technology
Rape, crucifer is full of nutrition, and wherein Vitamin C content is high, is planted in the range of Asia extensive.
Probenazole is a kind of benzimidazoles derivative, belongs to absorbability wide-spectrum bactericide, is widely used in sclerotiniose, the downy mildew of rape
Disease, black rot prevention, drug effect is 6 months, Long Term Contact or eats by mistake and easily triggers cancerous swelling, blood strangury disease and immune system disorderly
Disorderly, the physical and mental health of consumer is seriously affected.(GB 2763-2012) is provided according to China's Pesticide maximum residue limit,
The residual quantity of probenazole in the oil must not exceed 1mg/kg.At present, probenazole Pesticides Testing method mainly has gas chromatography-mass spectrum
Method, liquid chromatography etc., these methods have the characteristics that accurate, high sensitivity, but complex pretreatment, of high cost, detection time
Long, detecting instrument costliness, it is difficult to meet extensive online, low cost, the testing requirements of high speed.
Raman spectrum (Raman spectra), is a kind of scattering spectrum, and Raman spectrum analysis method is to be based on India scientist
C.V. the Raman scattering effect that Raman (Raman) is found, its principle are：Pair scattering spectrum different from incident light frequency carries out
Information in terms of analyzing to obtain molecular vibration, rotation.Surface enhanced Raman spectroscopy (surface-enhanced Raman
Spectroscopy, SERS) technology refers to Molecular Adsorption to the surface or molten of some nanoscale rough metals (such as gold, silver, copper)
In glue, make the Raman signal of adsorbent into the enhancing of geometry multiple.SERS technologies can be realized to micro-example and monomolecular
Quick detection, meanwhile, and a kind of Fingerprint of high sensitivity, there is instrument to be easy to carry about with one, is easy to operate, pre-treatment convenience
The features such as.In recent years, lot of domestic and international scholar applies Surface enhanced Raman spectroscopy to micro-example and monomolecular quick detection
Studied.
Surface enhanced Raman spectroscopy technology refers to Molecular Adsorption to the table of some nanoscale rough metals (such as gold, silver, copper)
In face or colloidal sol, the Raman signal of adsorbent is set to be adapted to carry out the remains of pesticide in agricultural product into the enhancing of geometry multiple
Rapid screening.Chetan etc. makes elargol capillary column and carried out post detection to orange juice, minimum dense with reference to SERS technologies, detection
Spend for 50 μ g/L.Li et al. is research pair with organophosphorus pesticide (thimet and Entex) using Nano silver grain as enhancing substrate
As choosing 2cm2Apple epidermis simulates pesticide residue, is detected with SERS, chooses 728cm respectively-1And 1215cm-1For mesh
Mark peak and carry out quantitative analysis, the results showed that test limit is respectively 0.05 and 0.4mg/L.Open duckweed etc. and use Surface enhanced Raman spectroscopy
Detection technique combination Accelerated solvent extraction pre-treating method establishes the quick inspection of basic element of cell division 6-BA residuals in bean sprouts
Survey method.Kim etc. gathers the Surface enhanced Raman spectroscopy of solution to be measured under different pH value using benzimidazole as research object,
Raman spectra is belonged to.He etc. combines a quick and easy method using SERS technologies realizes apple surface thiophene bacterium
The detection of clever pesticide, detection time only need 10min.Although the studies above can be reached pair using Surface enhanced Raman spectroscopy technology
The purpose that pesticide molecular signal strengthens in agricultural product, but fail fully to remove the influence of stroma ground substance, the test limit of method needs
Further improve, accuracy, the stability of method are poor, its detection method is also to be further improved, in addition, on utilizing table
There is not been reported for the remaining research of probenazole bactericide agricultural chemicals in face enhancing Raman spectroscopy detection rape.
The content of the invention
The present invention provides a kind of surface Raman enhancement technology based on silver nanoparticle colloidal sol substrate quickly to detect thiophene in rape
The method of bacterium spirit pesticide, quick, cheap, easy, accurate detection scheme is provided for Detecting Pesticide.
A kind of surface Raman enhancement technology based on silver nanoparticle colloidal sol substrate quickly detects the side of probenazole pesticide in rape
Method, including：
Step 1, by the rape, acetonitrile and sodium chloride that are attached with probenazole pesticide after mixing, vortex is carried out successively and is shaken
Swing, centrifuge, obtain supernatant；
Step 2, ferriferrous oxide nano-particle is mixed with supernatant, and carries out vortex oscillation, centrifugation, filtering successively,
Obtain prepare liquid；
Step 3, silver nanoparticle colloidal sol substrate is prepared；
Step 4, by silver nanoparticle colloidal sol substrate, prepare liquid and sodium-chloride water solution after mixing, Raman spectrum is carried out to adopt
Collection；
Step 5, the residual quantity of probenazole pesticide in rape is calculated according to equation below：
Y=0.0011x-15.355
In formula：Y be probenazole pesticide residual quantity, unit mg/kg；
X is 780cm-1Locate Raman absorption peak intensity, unit a.u..
The present invention is using rape as carrier, using benzimidazole germicide pesticide probenazole as research object, using four oxidations
The fluorescence that three Fe nanometer particles remove the materials such as chlorophyll, protein, fat, carbohydrate influences, using SERS technologies pair
Probenazole is remained and is used for quickly detecting in rape, to realize that it is a kind of quick, cheap, easy, accurate that Detecting Pesticide provides
Detection scheme.
In order to ensure being fully dissolved out for probenazole, it is preferable that in step 1, be attached with the rape of probenazole pesticide, acetonitrile,
Amount ratio with sodium chloride is 2g：2mL：1g.In step 1, after vortex oscillation 1min, 5min is centrifuged under 4200r/min rotating speeds,
Obtain supernatant.
In order to ensure that ferriferrous oxide nano-particle removes the effect of the materials such as chlorophyll, protein, fat, carbohydrate
Fruit, it is preferable that in step 2, the amount ratio of ferriferrous oxide nano-particle and supernatant is 300mg：1mL.In step 2, it is vortexed
After vibrating 1min, 2min is centrifuged under 4200r/min rotating speeds, after the supernatant after centrifugation crosses 0.22 μm of organic filter membrane, is treated
Survey liquid.
Preferably, trisodium citrate heating reduction method for preparing silver Nano sol substrate of the step 3 using Lee-Meisel.
Silver nanoparticle colloidal sol substrate can also use other modes to prepare, and can play Raman spectrum humidification, and do not influence target quality testing
Survey.
Preferably, in step 4, the mass fraction of sodium-chloride water solution is 1%, silver nanoparticle colloidal sol substrate, prepare liquid and
The volume ratio of sodium-chloride water solution is 5：1：1.
Preferably, in step 4, the parameter of Raman spectrum collection is arranged to：Excitation wavelength 785nm, power 200mW, sweeps
Retouch 200~3300cm of scope-1, optical resolution 2cm-1, time of integration 10s, integrates 3 times and takes average spectral value.
Compared with prior art, the present invention has the following advantages：
(1) surface Raman enhancement technology is applied to the detection of probenazole pesticide residue in rape first, is compared to biography
In the rape of system for the detection of probenazole pesticide residue, accuracy of detection is high, and the time is short, easy to operate；
(2) the PLS models of the actual value of probenazole pesticide residue in Raman absorption peak intensity and rape are established, quantitative detection
Probenazole pesticide residue.
Brief description of the drawings
Fig. 1 is that the surface Raman enhancement technology of the invention based on silver nanoparticle colloidal sol substrate quickly detects probenazole agriculture in rape
The flow chart of the method for medicine；
Fig. 2 is probenazole schematic arrangement；
Fig. 3 is the micrograph of the silver nanoparticle substrate of TEM transmission electron microscopes characterization；
Fig. 4 is the Surface enhanced Raman spectroscopy of probenazole pesticide residue in rape；
Fig. 5 is the Surface enhanced Raman spectroscopy of the first derivative processing of probenazole pesticide residue in rape；
Fig. 6 is the scatter diagram between predicted value and measured value after first derivative processing, wherein (a) collects for modeling, (b) is
Forecast set；
Fig. 7 is the linear regression graph of probenazole pesticide concentration in Raman absorption peak and rape.
Embodiment
Below in conjunction with the accompanying drawings, rape is quickly detected to surface Raman enhancement technology of the present invention based on silver nanoparticle colloidal sol substrate
The method of middle probenazole pesticide is described in detail.
As shown in Figure 1, a kind of surface Raman enhancement technology based on silver nanoparticle colloidal sol substrate quickly detects thiophene bacterium in rape
The method of clever pesticide, including：
Step 1,10g rapes sample is weighed in 50mL centrifuge tubes, 10mL acetonitriles, 5g sodium chloride is sequentially added, after shaking up
Vortex oscillation mixes 1min, then centrifuges 5min under 4200r/min rotating speeds, obtains green supernatant.
Step 2,2mL green supernatants are taken to be put into the 15mL centrifuge tubes equipped with 600mg ferriferrous oxide nano-particles, whirlpool
After rotation vibration mixing 1min, 2min is centrifuged under 4200r/min rotating speeds, after supernatant crosses 0.22 μm of organic filter membrane, as to be measured
Liquid is spare.
In this step rape Determination of Chlorophyll, protein, fat, carbohydrate are removed using ferriferrous oxide nano-particle
Fluorescence Deng material influences.
Step 3, silver nanoparticle colloidal sol substrate is prepared, concrete operations are as follows：
Reduction method is heated using the trisodium citrate of Lee-Meisel, by certain density silver nitrate solution (36mg nitric acid
Silver is dissolved in 200mL ultra-pure waters) pour into flask, it is placed in constant temperature blender with magnetic force, high temperature was heated rapidly to seethe with excitement, at 2 minutes
Mass fraction is inside progressively instilled as 1% citric acid three sodium solution (60mg trisodium citrates are dissolved in 6mL ultra-pure waters), while with
The rotating speed stirring of 200r/min, in the process, by transparent change light brown, celadon liquid slowly is obtained after reacting 25min for solution
Body.After room temperature cooling, above-mentioned appropriate fulmargin is poured into centrifuge tube, a small amount of supernatant, then past centrifugation are outwelled after centrifugation
Appropriate ultra-pure water is added in pipe, is mixed with sonic oscillation, elargol is kept in dark place after repeatedly purification.TEM transmission electron microscopes characterization
The micrograph of silver nanoparticle substrate is as shown in Figure 3.
Step 4, the silver nanoparticle substrate of 500 μ L steps 3 preparation is sequentially added in 2mL quartz bottles, prepared by 100 μ L steps 2
Prepare liquid, 100 μ L mass fractions be 1% sodium-chloride water solution, be put into supporting liquid sample pool collection SERS spectra.
Before gathering Raman spectrum data, rectified an instrument using acetonitrile.
The parameter of Raman spectrum collection is arranged to：Excitation wavelength 785nm, power 200mW, 200~3300cm of scanning range-1, optical resolution 2cm-1, time of integration 10s, integrates 3 times and takes average spectral value.
Step 5, the residual quantity of probenazole pesticide in rape is calculated according to equation below：
Y=0.0011x-15.355
In formula：Y is the residual quantity of probenazole pesticide, and unit is that (kg is the weight of rape to mg/kg, and mg is probenazole pesticide
Residual mass)；
X is 780cm-1Locate Raman absorption peak intensity, unit a.u..
Formula to establish process as follows：
(1) probenazole standard solution is prepared
1000mg probenazole standard items are dissolved in 1000mL acetonitriles, the probenazole standard solution of 1000mg/L is made into, puts
Stored in 4 DEG C of light protected environments, probenazole schematic arrangement is as shown in Figure 2.
(2) probenazole pesticide residue in rape is simulated
Choose without pesticide rape, the probenazole standard solution of the 1000mg/L in step 1 is diluted to the thiophene of various concentrations
Bacterium spirit solution is sprayed, and concentration range is 0~100mg/L, concentration gradient 1mg/L, when simulation nature sprinkling 24 is small after, adopt
Rape is plucked as experiment sample.
Experiment sample amounts to 101, wherein 70 sample composition modeling collection, remaining 31 sample composition forecast set.
(3) sample to be tested is prepared
Experiment sample is handled using the method for step 1 and step 2, obtains prepare liquid.
(4) silver nanoparticle colloidal sol substrate is prepared using the method for step 3, tests to obtain table using the parameter and method of step 4
Face strengthens Raman spectrum, and for Surface enhanced Raman spectroscopy as shown in figure 4, x-axis is wave number in Fig. 4, y-axis is Raman absorption peak intensity.
(5) actual value of probenazole pesticide residue in high performance liquid chromatography detection rape is used
Prepare liquid prepared by 2mL steps (3) is added equipped with 150mg magnesium sulfate, 50mg N- propyl group ethylenediamine (i.e.
Primary secondary amine, abbreviation PSA), in the 15mL centrifuge tubes of 10mg Graphons and 50mgC18 chromatographic columns,
After vortex oscillation mixing 2min, 5min is centrifuged under 4200r/min rotating speeds, takes supernatant 1mL in 10mL teat glasses, nitrogen
After drying, 1mL ethyl acetate is added, vortex oscillation, crosses 0.22 μm of organic filter membrane, be positioned in 2mL sample injection bottles, utilize efficient liquid
The actual value of probenazole pesticide residue, test result are as shown in table 1 in phase chromatography detection rape.
Table 1
(6) Pretreated spectra
The initial surface enhancing Raman spectrum gathered using preprocessing procedures to step (4) is handled, using one
Rank derivation pre-processes original spectral data, eliminates the interference of needle position misalignment, random noise and background.
Surface enhanced Raman spectroscopy in rape after the first derivative processing of probenazole pesticide residue is as shown in Figure 5.In Fig. 5
X-axis is wave number, and y-axis is Raman absorption peak intensity.
(7) actual value of probenazole pesticide in Raman spectrum absworption peak and rape is established using partial least-square regression method
Relationship model
Fig. 6 is modeling collection and the scatter diagram between the predicted value and measured value of forecast set sample after first derivative processing.Fig. 6
Middle x-axis is wave number, and y-axis is Raman absorption peak intensity.
Relationship model is the formula employed in step 5, and the linear regression graph of formula is as shown in fig. 7, the correlation of formula
Coefficients R2=0.9667.
According to the disclosure and teachings of the above specification, those skilled in the art in the invention can also be to above-mentioned embodiment party
Formula carries out appropriate change and modification.Therefore, the invention is not limited in embodiment disclosed and described above, to this
Some modifications and changes of invention should also be as falling into the scope of the claims of the present invention.In addition, although this specification
In used some specific terms, but these terms are merely for convenience of description, do not limit the present invention in any way.
Claims (8)
1. a kind of method that surface Raman enhancement technology based on silver nanoparticle colloidal sol substrate quickly detects probenazole pesticide in rape,
It is characterised in that it includes：
Step 1, the rape, acetonitrile and sodium chloride of probenazole pesticide will be attached with after mixing, carry out successively vortex oscillation,
Centrifugation, obtains supernatant；
Step 2, ferriferrous oxide nano-particle is mixed with supernatant, and carries out vortex oscillation, centrifugation, filtering successively, obtained
Prepare liquid；
Step 3, silver nanoparticle colloidal sol substrate is prepared；
Step 4, by silver nanoparticle colloidal sol substrate, prepare liquid and sodium-chloride water solution after mixing, Raman spectrum collection is carried out；
Step 5, the residual quantity of probenazole pesticide in rape is calculated according to equation below：
Y=0.0011x-15.355
In formula：Y be probenazole pesticide residual quantity, unit mg/kg；
X is 780cm-1Locate Raman absorption peak intensity, unit a.u..
2. the surface Raman enhancement technology as claimed in claim 1 based on silver nanoparticle colloidal sol substrate quickly detects thiophene bacterium in rape
The method of clever pesticide, it is characterised in that in step 1, be attached with the amount ratio of the rape of probenazole pesticide, acetonitrile and sodium chloride
For 2g：2mL：1g.
3. the surface Raman enhancement technology as claimed in claim 1 based on silver nanoparticle colloidal sol substrate quickly detects thiophene bacterium in rape
The method of clever pesticide, it is characterised in that in step 1, after vortex oscillation 1min, centrifuge 5min under 4200r/min rotating speeds, obtain
Supernatant.
4. the surface Raman enhancement technology as claimed in claim 1 based on silver nanoparticle colloidal sol substrate quickly detects thiophene bacterium in rape
The method of clever pesticide, it is characterised in that in step 2, the amount ratio of ferriferrous oxide nano-particle and supernatant is 300mg：
1mL。
5. the surface Raman enhancement technology as claimed in claim 1 based on silver nanoparticle colloidal sol substrate quickly detects thiophene bacterium in rape
The method of clever pesticide, it is characterised in that in step 2, after vortex oscillation 1min, 2min is centrifuged under 4200r/min rotating speeds, is centrifuged
After supernatant afterwards crosses 0.22 μm of organic filter membrane, prepare liquid is obtained.
6. the surface Raman enhancement technology as claimed in claim 1 based on silver nanoparticle colloidal sol substrate quickly detects thiophene bacterium in rape
The method of clever pesticide, it is characterised in that step 3 is molten using the trisodium citrate heating reduction method for preparing silver nanometer of Lee-Meisel
Matrix bottom.
7. the surface Raman enhancement technology as claimed in claim 1 based on silver nanoparticle colloidal sol substrate quickly detects thiophene bacterium in rape
The method of clever pesticide, it is characterised in that in step 4, the mass fraction of sodium-chloride water solution is 1%, and silver nanoparticle colloidal sol substrate, treat
The volume ratio for surveying liquid and sodium-chloride water solution is 5：1：1.
8. the surface Raman enhancement technology as claimed in claim 1 based on silver nanoparticle colloidal sol substrate quickly detects thiophene bacterium in rape
The method of clever pesticide, it is characterised in that in step 4, the parameter of Raman spectrum collection is arranged to：Excitation wavelength 785nm, power
200mW, 200~3300cm of scanning range-1, optical resolution 2cm-1, time of integration 10s, integrates 3 times and takes average spectral value.
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| CN110441282A (en) * | 2019-07-19 | 2019-11-12 | 盐城工学院 | A kind of Pesticide Residues in Tea detection method based on Surface enhanced Raman spectroscopy and papery micro-fluidic chip |
| CN110907429A (en) * | 2019-12-31 | 2020-03-24 | 广东海洋大学 | Surface enhanced Raman spectroscopy detection method for micro/nano plastic |
| CN111060489A (en) * | 2019-12-25 | 2020-04-24 | 安徽中科赛飞尔科技有限公司 | SERS detection method for dazomet or bromadiolone in edible oil |
| CN111289491A (en) * | 2020-03-17 | 2020-06-16 | 浙江中烟工业有限责任公司 | Method for detecting triadimefon and triadimenol in tobacco based on surface enhanced Raman spectroscopy |
| CN111735805A (en) * | 2020-06-19 | 2020-10-02 | 上海如海光电科技有限公司 | Raman rapid detection method for fenoxycarb in food and fruit |
| CN112461808A (en) * | 2019-09-06 | 2021-03-09 | 苏州市农产品质量安全监测中心 | Detection method and kit for detecting carbendazim in agricultural products |
| CN112798704A (en) * | 2020-12-28 | 2021-05-14 | 江西省食品检验检测研究院（江西国家果蔬产品及加工食品质量监督检验中心） | Method for detecting pesticide residue in plant-derived agricultural products |
| CN113008870A (en) * | 2021-03-23 | 2021-06-22 | 浙江大学山东(临沂)现代农业研究院 | Microscopic detection method for sclerotinia sclerotiorum infection of rape stalks based on microscopic Raman spectrum |
| CN113960013A (en) * | 2021-11-08 | 2022-01-21 | 北京同仁堂健康（大连）海洋食品有限公司 | Method for detecting nitrofuran drug metabolites based on Br-assisted SERS |
| CN117554354A (en) * | 2024-01-12 | 2024-02-13 | 江苏诺恩作物科学股份有限公司 | Spectral detection analysis method and processing system for thiabendazole wastewater sample |
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| CN110441282A (en) * | 2019-07-19 | 2019-11-12 | 盐城工学院 | A kind of Pesticide Residues in Tea detection method based on Surface enhanced Raman spectroscopy and papery micro-fluidic chip |
| CN112461808A (en) * | 2019-09-06 | 2021-03-09 | 苏州市农产品质量安全监测中心 | Detection method and kit for detecting carbendazim in agricultural products |
| CN111060489A (en) * | 2019-12-25 | 2020-04-24 | 安徽中科赛飞尔科技有限公司 | SERS detection method for dazomet or bromadiolone in edible oil |
| CN110907429A (en) * | 2019-12-31 | 2020-03-24 | 广东海洋大学 | Surface enhanced Raman spectroscopy detection method for micro/nano plastic |
| CN110907429B (en) * | 2019-12-31 | 2023-09-19 | 广东海洋大学 | Surface enhanced Raman spectrum detection method for micro/nano plastic |
| CN111289491B (en) * | 2020-03-17 | 2023-03-10 | 浙江中烟工业有限责任公司 | Method for detecting triadimefon and triadimenol in tobacco based on surface enhanced Raman spectroscopy |
| CN111289491A (en) * | 2020-03-17 | 2020-06-16 | 浙江中烟工业有限责任公司 | Method for detecting triadimefon and triadimenol in tobacco based on surface enhanced Raman spectroscopy |
| CN111735805A (en) * | 2020-06-19 | 2020-10-02 | 上海如海光电科技有限公司 | Raman rapid detection method for fenoxycarb in food and fruit |
| CN112798704A (en) * | 2020-12-28 | 2021-05-14 | 江西省食品检验检测研究院（江西国家果蔬产品及加工食品质量监督检验中心） | Method for detecting pesticide residue in plant-derived agricultural products |
| CN113008870A (en) * | 2021-03-23 | 2021-06-22 | 浙江大学山东(临沂)现代农业研究院 | Microscopic detection method for sclerotinia sclerotiorum infection of rape stalks based on microscopic Raman spectrum |
| CN113960013A (en) * | 2021-11-08 | 2022-01-21 | 北京同仁堂健康（大连）海洋食品有限公司 | Method for detecting nitrofuran drug metabolites based on Br-assisted SERS |
| CN117554354A (en) * | 2024-01-12 | 2024-02-13 | 江苏诺恩作物科学股份有限公司 | Spectral detection analysis method and processing system for thiabendazole wastewater sample |
| CN117554354B (en) * | 2024-01-12 | 2024-03-15 | 江苏诺恩作物科学股份有限公司 | Spectral detection analysis method and processing system for thiabendazole wastewater sample |
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