CN111289491A - Method for detecting triadimefon and triadimenol in tobacco based on surface enhanced Raman spectroscopy - Google Patents
Method for detecting triadimefon and triadimenol in tobacco based on surface enhanced Raman spectroscopy Download PDFInfo
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Abstract
The invention discloses a method for detecting triadimefon and triadimenol in tobacco based on surface-enhanced Raman spectroscopy, which comprises the following steps: (1) soaking a tobacco sample in pure water, adding an organic reagent, performing vortex oscillation and centrifugation, taking supernatant liquid, sequentially adding a purifying agent and hydrochloric acid into a new centrifugal tube, performing vortex oscillation and centrifugation, taking an organic layer, drying, adding pure water, and redissolving to obtain a sample liquid to be detected; (2) uniformly mixing the sample solution to be detected with an agglomeration agent and a nanogold sol, and collecting a surface enhanced Raman spectrum of the mixed solution by using a portable Raman spectrometer; (3) analyzing the measured surface enhanced Raman spectrogram to obtain the detection results of triadimefon and triadimenol. The detection method does not need to depend on large-scale equipment, adopts a portable Raman spectrometer combined with a simple and rapid pretreatment technology, has low detection cost, and is suitable for rapid screening of field batch samples.
Description
Technical Field
The invention belongs to the field of detection of triadimefon and triadimenol, and particularly relates to a method for detecting triadimefon and triadimenol in tobacco based on surface-enhanced Raman spectroscopy.
Background
Triazolone is a broad-spectrum triazole bactericide with high efficiency, low toxicity and strong systemic property, is mainly used for preventing and treating plant fungal viruses such as rust disease, powdery mildew and the like, is widely used in agricultural production, and registered crops comprise tobacco, wheat, rice, corn, peanut, tomato, apple trees and the like. Triadimenol is a metabolite of triadimefon, which has a higher bactericidal activity than triadimefon and has been registered as a single agent for use on a variety of crops. Triazolones and triadimenol have been found to produce neurological, reproductive and genotoxicity, as well as carcinogenicity and teratogenicity. In order to ensure scientific and reasonable use of two pesticides, the maximum residual limit of the pesticides in food and crops is clearly defined in many countries such as China, and the maximum residual limit standard of the pesticides in tobacco leaves is correspondingly released by China tobacco general company. Therefore, the method has important significance for detecting residual quantity of triadimefon and triadimenol, especially for rapid detection.
At present, methods for detecting triazolone and triadimenol in tobacco and tobacco products mainly comprise gas chromatography, liquid chromatography-tandem mass spectrometry, gas chromatography-tandem mass spectrometry and the like. For example, the invention patent CN103698462B discloses a method for detecting pesticide residues in tobacco by liquid chromatography-tandem mass spectrometry, and the invention patent CN104458947B discloses a method for detecting pesticide residues in tobacco or tobacco products by gas chromatography-mass spectrometry, which have the advantages of good accuracy, high sensitivity, capability of simultaneously detecting various pesticide residues, and the like, but the required instruments and equipment are expensive, professional technicians are required, the pretreatment of samples is complicated, and the detection time is long.
The Raman spectroscopy (Raman spectra) analysis method is an analysis method which is used for analyzing a scattering spectrum with different incident light frequencies to obtain molecular vibration and rotation information and is applied to molecular structure research based on a Raman scattering effect found by indian scientists c.v. Raman (Raman). However, most molecules usually have extremely small scattering cross sections, so that the raman scattering signals are very weak, and a clear raman spectrum can be obtained only by a high concentration, thereby greatly limiting the practical application of the raman spectrum in the level analysis of low-concentration target substances. The Surface Enhanced Raman Scattering (SERS) effect discovered in the 70's of the 20 th century can enhance Raman signals, overcomes the disadvantage of weak conventional Raman signals, can obtain high-sensitivity structural information at the level of low-concentration substances to be detected, even single molecules, has the characteristics of high analysis speed, low detection cost, simplicity and convenience in operation and the like, is a rapidly-developed scattering spectrum technology, and is one of the research hotspots at the present stage. Currently, the SERS has attracted more and more attention for analyzing and detecting pesticide residues. However, the complexity of the tobacco system and the matching requirement of the sample pretreatment with rapidness, convenience and low cost bring certain challenges to the SERS rapid detection of the pesticide residue of the tobacco sample.
Disclosure of Invention
The invention aims to provide a method for detecting triazolone and triadimenol in tobacco based on surface-enhanced Raman spectroscopy, which adopts a portable Raman spectrometer and combines a tobacco sample pretreatment technology with rapidness, simplicity, convenience and low cost, and is suitable for rapid screening of field batch samples.
The technical scheme of the invention is as follows:
a method for detecting triadimefon and triadimenol in tobacco based on surface enhanced Raman spectroscopy comprises the following steps:
(1) soaking a tobacco sample in pure water, adding an organic reagent, performing vortex oscillation and centrifugation, taking supernatant liquid, sequentially adding a purifying agent and hydrochloric acid into a new centrifugal tube, performing vortex oscillation and centrifugation, taking an organic layer, drying, adding pure water, and redissolving to obtain a sample liquid to be detected;
(2) uniformly mixing the sample solution to be detected with an agglomeration agent and a nanogold sol, and collecting a surface enhanced Raman spectrum of the mixed solution by using a portable Raman spectrometer;
(3) analyzing the measured surface enhanced Raman spectrogram to obtain the detection results of triadimefon and triadimenol.
In order to ensure that the triazolone and the triadimenol are sufficiently dissolved, in the step (1), the tobacco samples are prepared according to the proportion of 1: 5-20, mixing with pure water, and mixing with a tobacco sample 1: 2-10, adding organic solvent, oscillating for 0.5-5 min by vortex, and centrifuging for 1-2 min at 8000-10000 r/min. Preferably, the mixing ratio of the tobacco sample, the pure water and the organic solvent is 1: 7-13: 4 to 7. The organic solvent comprises one of ethyl acetate, cyclohexane, petroleum ether, dichloromethane, diethyl ether or trichloromethane.
In order to remove the interference of impurities such as pigments, phenols, organic acids and the like in the tobacco, in the step (1), polyamide is used as a purifying agent, the dosage of the polyamide in each gram of tobacco sample is 0-0.5 g, the dosage of 1mol/L hydrochloric acid is 10-200 mu L, vortex oscillation is carried out for 0.5-1.5 min, and centrifugation is carried out for 1-2 min at 8000-10000 r/min. Preferably, the polyamide is used in an amount of 0.05 to 0.3g, and 1mol/L hydrochloric acid is added in an amount of 40 to 80. mu.L.
In order to adjust the ionic strength of the system and enable the gold nanoparticles to obtain the optimal reinforcing capacity, in the step (2), the agglomerating agent comprises one or a mixture of sodium chloride, potassium chloride, sodium sulfate, magnesium sulfate, hydrochloric acid and potassium iodide, and preferably, the agglomerating agent is a mixture of 1mol/L hydrochloric acid and 1mol/L sodium sulfate in a volume ratio of 1: 2-5 mixing.
In the step (2), the particle size of the nano gold sol particles is 20-150 nm, the enhancement capability, the synthesis controllability of the morphology and the size, the system stability and the like of the nano gold sol in a comprehensive system are integrated, and preferably, the particle size of the nano gold sol particles is 55-75 nm.
In the step (2), the volume ratio of the sample liquid to be detected, the agglomeration agent and the nanogold sol is 1: 0.1-0.6: 0.8 to 1.5. Preferably, the volume ratio of the sample liquid to be detected, the agglomerating agent and the nanogold sol is 1: 0.1-0.3: 0.9 to 1.2.
In the step (2), the parameters of the portable Raman spectrometer are 785nm of an excitation light source, the excitation power is 50-500 mW, the scanning time is 100-10000 ms, preferably, the excitation power is 500mW, and the scanning time is 5000 ms.
In the step (3), the collected surface enhanced Raman spectrum of the sample is compared with the surface enhanced Raman spectrum of the triadimefon and triadimenol standard, and whether the sample contains triadimefon and triadimenol can be judged according to the characteristic peak. Wherein, the surface enhanced Raman characteristic peak of the triadimenol is as follows: 536cm-1、632cm-1、745cm-1、862cm-1、1012cm-1、1091cm-1、1221cm-1(ii) a Surface enhanced raman characteristic peaks of triadimefon: 589cm-1、641cm-1、830cm-1、987cm-1、1091cm-1、1221cm-1。
Compared with the prior art, the invention has the beneficial effects that:
the method for detecting triadimefon and triadimenol in tobacco based on the surface-enhanced Raman spectrum has the advantages of simple pretreatment steps of tobacco samples, convenience in operation, rapidness, high efficiency, low cost, small organic reagent consumption, no dependence on large-scale equipment and suitability for rapid screening of field batch samples.
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In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to these drawings without creative efforts.
FIG. 1 is a scanning electron microscope image of gold nanoparticles;
FIG. 2 shows the surface enhanced Raman spectrum and the solid Raman spectrum of a triazolone standard solution, wherein a, b, c, d and e are respectively 0, 0.01, 0.05, 0.1 and 1mg/L of the triazolone standard solution, and f is the triazolone solid Raman spectrum;
FIG. 3 is the surface enhanced Raman spectrum and the solid Raman spectrum of the triadimenol standard solution, wherein a, b, c and d are the surface enhanced Raman spectrum of the triadimenol standard solution of 0, 0.05, 0.1 and 1mg/L respectively, and e is the solid Raman spectrum of the triadimenol;
FIG. 4 is a surface enhanced Raman spectrum of triadimenol in a tobacco sample, wherein a is a surface enhanced Raman spectrum of a tobacco blank sample extract, b, c and d are surface enhanced Raman spectra of tobacco sample extract with triadimenol of 1, 5 and 10 μ g/g, respectively, and e is a surface enhanced Raman spectrum of a triadimenol standard;
FIG. 5 is a surface enhanced Raman spectrum of triazolone in a tobacco sample, wherein a is a surface enhanced Raman spectrum of a tobacco blank sample extract, b, c and d are respectively surface enhanced Raman spectra of triazolone-added tobacco sample extract labeled with 1, 5 and 10 μ g/g, and e is a surface enhanced Raman spectrum of a triazolone standard substance;
FIG. 6 is a surface-enhanced Raman spectrum of an extract of a tobacco blank sample in the presence of both triadimenol and triadimefon, wherein a is a surface-enhanced Raman spectrum of the extract of the tobacco blank sample, b, c and d are respectively a surface-enhanced Raman spectrum of an extract of the tobacco sample labeled with 5. mu.g/g triadimenol and 1. mu.g/g triadimenol, labeled with 1. mu.g/g triadimenol and 5. mu.g/g triadimefon, labeled with 5. mu.g/g triadimenol and 5. mu.g/g triadimefon, and e is a surface-enhanced Raman spectrum of a mixed standard of triadimenol and triadimefon.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention will be further described in detail with reference to the accompanying drawings and examples. It should be understood that the detailed description and specific examples, while indicating the scope of the invention, are intended for purposes of illustration only and are not intended to limit the scope of the invention.
The preparation of the nano gold sol comprises the following steps:
200mL of a 0.01 wt% chloroauric acid solution was boiled, and then 1.5mL of a sodium citrate solution (1 wt% concentration) was rapidly added thereto at a time to change the solution to reddish brown in about 3min, and the mixture was kept boiling for 30min and cooled for use. The prepared gold nanoparticles were characterized by scanning electron microscopy, and the results are shown in fig. 1. The gold nanoparticles are spherical with uniform size and the diameter is about 70 nanometers.
Raman spectrum collection of triadimefon and triadimenol standard
Respectively placing standard triadimefon and triadimenol solid powder on a clean glass slide, flattening, and performing Raman spectrum collection on a portable Raman spectrometer to obtain Raman spectra shown in figures 2 and 3.
Preparing standard aqueous solutions (1mg/L, 0.1mg/L, 0.05mg/L, 0.01mg/L and 0mg/L) of triadimefon and triadimenol with different concentrations respectively. And (3) adding 50 mu L of an agglomeration agent (1mol/L sodium sulfate and 1mol/L hydrochloric acid are mixed according to a volume ratio of 4: 1) into 200 mu L of standard solution, adding 200 mu L of gold nano sol, and collecting a Raman spectrum by using a portable Raman spectrometer under the conditions of 785nm of exciting light, 500mW of laser power and 5000ms of scanning time. The result shows that the detection concentration of the standard solution of triadimefon and triadimenol can reach 0.05mg/L by the detection method provided by the invention, as shown in fig. 2 and 3.
Detection of triadimenol in tobacco samples
Adding a standard solution of triadimenol into 1g of tobacco blank samples according to the ratio of 1 mug/g, 5 mug/g and 10 mug/g respectively, after the standard solution is completely absorbed and dried, sequentially adding 10mL of pure water and 5mL of cyclohexane, carrying out vortex oscillation for 2min, centrifuging at 8000r/min for 2min, taking 4mL of upper-layer liquid into a new centrifuge tube, adding 0.1g of polyamide and 50 muL of 1mol/L hydrochloric acid, oscillating for 1min, centrifuging at 8000r/min for 2min, taking 3mL of upper-layer liquid nitrogen, carrying out air blowing drying, and adding 400 muL of pure water to redissolve to obtain a sample solution to be detected. Taking 250 mu L of sample liquid to be detected, adding 50 mu L of agglomeration agent (1mol/L sodium sulfate and 1mol/L hydrochloric acid are mixed according to the volume ratio of 3: 1), adding 250 mu L of nano gold sol, and collecting the Raman spectrum of the sample by using a portable Raman spectrometer under the conditions of 785nm of exciting light, 500mW of laser power and 5000ms of scanning time. The result shows that the detection concentration of the standard triadimenol in the tobacco sample by the detection method provided by the invention can reach 1 mug/g, as shown in figure 4.
Detection of triazolone in tobacco sample
Adding standard solutions of triazolone into 1g of tobacco blank samples according to the proportion of 1 mug/g, 5 mug/g and 10 mug/g respectively, after the standard solutions are completely absorbed and dried, sequentially adding 10mL of pure water and 5mL of petroleum ether, carrying out vortex oscillation for 3min, centrifuging for 2min at 8000r/min, taking 4mL of upper-layer liquid into a new centrifugal tube, adding 0.15g of polyamide and 60 mug of 1mol/L hydrochloric acid, oscillating for 0.5min, centrifuging for 1min at 10000r/min, taking 3mL of upper-layer liquid nitrogen, carrying out air blowing drying, adding 400 mug of pure water, and redissolving to obtain a sample solution to be detected. Adding 50 mu L of an agglomeration agent (1mol/L sodium sulfate and 1mol/L hydrochloric acid are mixed according to the volume ratio of 4: 1) into 200 mu L of sample liquid to be detected, adding 200 mu L of nano gold sol, and collecting the Raman spectrum of the sample by using a portable Raman spectrometer under the conditions of 785nm of exciting light, 500mW of laser power and 5000ms of scanning time. The result shows that the detection concentration of the labeled triazolone in the tobacco sample by the detection method provided by the invention can reach 1 mug/g, as shown in figure 5.
Simultaneous detection of triadimefon and triadimenol in tobacco samples
Adding a standard solution of triazolone and triadimenol into a 1g tobacco blank sample, after the standard solution is completely absorbed and dried, sequentially adding 12mL of pure water and 6mL of petroleum ether, carrying out vortex oscillation for 5min, centrifuging at 10000r/min for 2min, taking 4mL of upper layer solution, adding 0.1g of polyamide and 50 mu L of hydrochloric acid at 1mol/L into a new centrifugal tube, oscillating for 1min, centrifuging at 8000r/min for 2min, taking 3mL of upper layer liquid nitrogen, air-drying, and adding 400 mu L of pure water for redissolving to obtain a sample solution to be detected. And adding 50 mu L of an agglomeration agent into 300 mu L of sample liquid to be detected, adding 300 mu L of nano gold sol, and collecting the Raman spectrum of the sample by using a portable Raman spectrometer under the conditions of 785nm of exciting light, 500mW of laser power and 5000ms of scanning time. The result shows that the detection method provided by the invention can realize the simultaneous detection of the labeled triadimefon and triadimenol in the tobacco sample, as shown in fig. 6.
The above-mentioned embodiments are intended to illustrate the technical solutions and advantages of the present invention, and it should be understood that the above-mentioned embodiments are only the most preferred embodiments of the present invention, and are not intended to limit the present invention, and any modifications, additions, equivalents, etc. made within the scope of the principles of the present invention should be included in the scope of the present invention.
Claims (10)
1. A method for detecting triadimefon and triadimenol in tobacco based on surface enhanced Raman spectroscopy comprises the following steps:
(1) soaking a tobacco sample in pure water, adding an organic reagent, performing vortex oscillation and centrifugation, taking supernatant liquid, sequentially adding a purifying agent and hydrochloric acid into a new centrifugal tube, performing vortex oscillation and centrifugation, taking an organic layer, drying, adding pure water, and redissolving to obtain a sample liquid to be detected;
(2) uniformly mixing the sample solution to be detected with an agglomeration agent and a nanogold sol, and collecting a surface enhanced Raman spectrum of the mixed solution by using a portable Raman spectrometer;
(3) analyzing the measured surface enhanced Raman spectrogram to obtain the detection results of triadimefon and triadimenol.
2. The method for detecting triazolone and triadimenol in tobacco based on surface enhanced raman spectroscopy according to claim 1, wherein in the step (1), the ratio of the triadimenol to the triadimenol in the tobacco sample is determined according to the following formula 1: 5-20, mixing with pure water, and mixing with a tobacco sample 1: 2-10, adding organic solvent, oscillating for 0.5-5 min by vortex, and centrifuging for 1-2 min at 8000-10000 r/min.
3. The method for detecting triazolone and triadimenol in tobacco based on surface enhanced raman spectroscopy according to claim 1, wherein the mixing ratio of the tobacco sample, pure water and the organic solvent is 1: 7-13: 4 to 7.
4. The method for detecting triadimefon and triadimenol in tobacco based on surface-enhanced raman spectroscopy of claim 1, wherein the organic solvent comprises one of ethyl acetate, cyclohexane, petroleum ether, dichloromethane, diethyl ether or chloroform.
5. The method for detecting triazolone and triadimenol in tobacco based on surface enhanced Raman spectroscopy according to claim 1, wherein in step (1), polyamide is used as a purifying agent, the amount of polyamide used per gram of tobacco sample is 0-0.5 g, and the amount of 1mol/L hydrochloric acid is 10-200 μ L.
6. The method for detecting triadimefon and triadimenol in tobacco based on surface enhanced Raman spectroscopy of claim 1, wherein the agglomeration agent comprises one or more of sodium chloride, potassium chloride, sodium sulfate, magnesium sulfate, hydrochloric acid and potassium iodide.
7. The method for detecting triazolone and triadimenol in tobacco based on surface enhanced raman spectroscopy of claim 1, wherein the agglomeration agent is 1mol/L hydrochloric acid and 1mol/L sodium sulfate in a volume ratio of 1: 2-5 mixing.
8. The method for detecting triazolone and triadimenol in tobacco based on surface-enhanced Raman spectroscopy according to claim 1, wherein the particle size of the nanogold sol particles is 20-150 nm.
9. The method for detecting triadimefon and triadimenol in tobacco based on surface-enhanced Raman spectroscopy according to claim 1, wherein in the step (2), the volume ratio of the sample liquid to be detected, the agglomerating agent and the nanogold sol is 1: 0.1-0.6: 0.8 to 1.5.
10. The method for detecting triazolone and triadimenol in tobacco based on surface-enhanced Raman spectroscopy according to claim 1, wherein in the step (2), the parameters of the portable Raman spectrometer are 785nm of an excitation light source, the excitation power is 50-500 mW, and the scanning time is 100-10000 ms.
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| CN112595702B (en) * | 2020-12-22 | 2024-06-11 | 云南省烟草质量监督检测站 | Method for rapidly detecting hexaconazole in tobacco by surface enhanced Raman scattering |
| CN113375997A (en) * | 2021-08-16 | 2021-09-10 | 赛默飞世尔（上海）仪器有限公司 | Method and product for detecting fentanyl compound |
| CN113375997B (en) * | 2021-08-16 | 2022-02-18 | 赛默飞世尔（上海）仪器有限公司 | Method and product for detecting fentanyl compound |
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