阅读说明：本技术 一种基于复合纳米材料的动物源性食品中克伦特罗的快速检测方法 (method for rapidly detecting clenbuterol in animal-derived food based on composite nano material ) 是由 程劼 苏晓鸥 王石 王培龙 王瑞国 于 2019-10-28 设计创作，主要内容包括：本发明公开了一种基于复合纳米材料的动物源性食品中克伦特罗的快速检测方法。首先制备二次深度氧化石墨烯负载纳米金ro-GO/AuNPs作为SERS基底；对待检测样品进行净化处理,获得待测溶液；向所述待测溶液加入ro-GO/AuNPs复合基底,并对其进行拉曼光谱扫描,采集光谱数据；针对采集的光谱数据,以△v＝1265,1474,1602cm<Sup>-1</Sup>处拉曼吸收作为克伦特罗的定性特征峰,并通过△v＝1474cm<Sup>-1</Sup>处拉曼吸收强度进行定量计算。上述方法具有快速、高灵敏、重复性好的特点,大大提高了检测效率。(The invention discloses a rapid detection method of clenbuterol in animal-derived foods based on composite nano materials, which comprises the steps of firstly preparing secondary deep graphene oxide loaded nano gold ro-GO/AuNPs as an SERS substrate, purifying a sample to be detected to obtain a solution to be detected, adding the ro-GO/AuNPs composite substrate into the solution to be detected, performing Raman spectrum scanning on the solution to be detected, acquiring spectral data, and regarding the acquired spectral data, adopting △ v 1265, 1474 and 1602cm ‑1 Raman absorption as the qualitative characteristic peak of clenbuterol and passing △ v-1474 cm ‑1 And (4) performing quantitative calculation on the Raman absorption intensity. The method has the characteristics of high speed, high sensitivity and good repeatability, and greatly improves the detection efficiency.)

1, detection method of clenbuterol in animal derived food based on composite nano material, characterized in that the method comprises:

step 1, firstly, preparing secondary deep graphene oxide loaded nano auro-GO/AuNPs as a surface enhanced Raman spectrum SERS enhanced composite substrate;

step 2, carrying out pretreatment on a clenbuterol CL sample in the animal-derived food to be detected to obtain a solution to be detected;

step 3, adding a ro-GO/AuNPs composite substrate into the solution to be detected, performing Raman scanning on the solution to be detected, and collecting spectral data in a corresponding range;

and 4, carrying out △ v-1265, 1474 and 1602cm on the acquired spectral data^-1(ii) process RamanAbsorption was taken as a characteristic qualitative peak for clenbuterol and passed △ v-1474 cm^-1And quantitatively calculating the Raman absorption intensity to realize the detection of the clenbuterol.

2. The method for detecting clenbuterol in animal derived food based on composite nanomaterial according to claim 1, wherein the process of step 1 specifically comprises:

firstly, mixing and heating 8mL of graphene oxide GO solution, 50mL of deionized water and 5.6mL of oxidant to prepare an excess solution;

adding 5mL of oxidant into part of the solution, and continuously heating and stirring to prepare a secondary deep graphene oxide ro-GO solution;

then adding citric acid aqueous solution and 1mL of chloroauric acid, and continuously heating and stirring to obtain the secondary deep graphene oxide loaded nano auro-GO/AuNPs.

3. The method for detecting clenbuterol in animal derived food based on composite nanomaterial according to claim 1, wherein the process of step 2 specifically comprises:

adding 10mL of dichloromethane and 5.0g of anhydrous sodium sulfate into 5.0g of the crushed sample to be detected for times of extraction;

transferring the extracted solvent into a centrifuge tube, adding 8mL of nitric acid aqueous solution, oscillating, extracting, centrifuging, and transferring part of upper layer solution;

then adding 20 mu L of NaOH aqueous solution and 800 mu L of dichloromethane for secondary reverse extraction, oscillating and centrifuging;

and then adding 800 mu L of nitric acid solution into the lower layer solution for third extraction, oscillating and centrifuging to obtain an upper layer solution which is the solution to be detected.

4. The method for detecting clenbuterol in animal derived food based on composite nanomaterial according to claim 1, wherein the process of step 3 specifically comprises:

adding 600 mu L of ro-GO/AuNPs composite substrate and 80uL of nitric acid into 10 mu L of solution to be detected, and oscillating for 60 s;

then carrying out Raman scanning on the Raman-scanned optical fiber, wherein the specific parameters are as follows: the spectrum scanning time is 10s, the scanning frequency is 2, the smoothing parameter is 1, the scanning power is 100mw, and the collection is 1000-2000cm^-1Spectral data within a range.

Technical Field

The invention relates to the technical field of surface-enhanced Raman spectroscopy application, in particular to a rapid detection method for clenbuterol in animal-derived foods based on composite nano materials.

Background

The common analysis method of CL in animal tissues is liquid chromatography-tandem mass spectrometry, the method is accurate, sensitive and reliable and is suitable for confirmed analysis in laboratories, but expensive instrument equipment and complex sample pretreatment steps make the method unsuitable for on-site rapid screening.

The Surface Enhanced Raman Spectroscopy (SERS) technology is taken as rapid analysis means, and is very concerned in recent years, under the synergistic effect based on physical 'hot spot' enhancement and charge transfer chemical enhancement, the Raman absorption displacement of a compound is Enhanced, trace analysis can be realized, the construction of an Enhanced substrate and efficient sample purification are the key for realizing rapid and high-sensitivity SERS analysis of a target compound in an actual sample, research reports are provided for the SERS analysis of clenbuterol, but the target substrate of the analysis is basically urine, in order to improve the sensitivity and selectivity of the analysis, most of literature reports are that enrichment of target CL molecules is realized based on the specific recognition of CL antibodies, but the preparation of the CL antibodies requires more complicated processes.

Disclosure of Invention

The invention aims to provide a rapid detection method for clenbuterol in animal-derived foods based on composite nano materials, which has the characteristics of rapidness, high sensitivity and good repeatability, and the detection sensitivity can reach 1ng g^-1The detection time is less than 10 min/sample.

The purpose of the invention is realized by the following technical scheme:

method for detecting clenbuterol in animal derived food based on composite nano material, comprising the following steps:

step 1, firstly, preparing secondary deep graphene oxide loaded nano auro-GO/AuNPs as a surface enhanced Raman spectrum SERS enhanced composite substrate;

step 2, carrying out pretreatment on a clenbuterol CL sample in the animal-derived food to be detected to obtain a solution to be detected;

step 3, adding a ro-GO/AuNPs composite substrate into the solution to be detected, performing Raman scanning on the solution to be detected, and collecting spectral data in a corresponding range;

and 4, carrying out △ v-1265, 1474 and 1602cm on the acquired spectral data^-1Raman absorption as characteristic qualitative peak of clenbuterol, and passing △ v-1474 cm^-1And quantitatively calculating the Raman absorption intensity to realize the detection of the clenbuterol.

The technical scheme provided by the invention shows that the method has the characteristics of high speed, high sensitivity and good repeatability, and the detection sensitivity can reach 1ng g^-1The detection time is less than 10 min/sample.

Drawings

In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings used in the description of the embodiments will be briefly introduced below, and it is obvious that the drawings in the following description are only embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings based on the drawings without creative efforts.

Fig. 1 is a schematic flow chart of a method for detecting clenbuterol in an animal-derived food based on a composite nanomaterial provided in an embodiment of the present invention;

fig. 2 is a schematic diagram of characteristic qualitative peaks of clenbuterol provided in an embodiment of the present invention;

fig. 3 is a clenbuterol quantitation curve provided by an embodiment of the present invention.

Detailed Description

The technical solutions in the embodiments of the present invention are clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only partial embodiments of the present invention , rather than all embodiments.

The following will describe the embodiment of the present invention in further steps with reference to the accompanying drawings, and as shown in fig. 1, is a schematic flow chart of the method for detecting clenbuterol in animal-derived food based on composite nanomaterial provided in the embodiment of the present invention, the method mainly includes:

step 1, firstly, preparing secondary deep graphene oxide loaded nano-gold ro-GO/AuNPs (twin-oxide graphene oxide/gold nanoparticles) as a surface enhanced Raman spectrum SERS enhanced composite substrate;

in the step, the preparation process specifically comprises the following steps:

firstly, mixing and heating 8mL of graphene oxide GO solution, 50mL of deionized water and 5.6mL of oxidant to prepare an excess solution;

adding 5mL of oxidant into part of the solution, and continuously heating and stirring to prepare a secondary deep graphene oxide ro-GO solution;

then adding citric acid aqueous solution and 1mL of chloroauric acid, and continuously heating and stirring to obtain the secondary deep graphene oxide loaded nano auro-GO/AuNPs.

The obtained ro-GO/AuNPs can be stored at 4 ℃ for later use.

The preparation process can be known as follows: through two times of deep oxidation to graphene oxide GO, the load concentration of GO to AuNPs can be improved, and therefore the SERS activity and stability of the ro-GO/AuNPs serving as a composite substrate are improved.

Step 2, carrying out pretreatment on a clenbuterol CL sample in the animal-derived food to be detected to obtain a solution to be detected;

in the step, the specific process is as follows:

firstly, adding 10mL of dichloromethane and 5.0g of anhydrous sodium sulfate into 5.0g of a crushed sample to be detected to perform times of extraction;

transferring the extracted solvent into a centrifuge tube, adding 8mL of nitric acid aqueous solution, oscillating, extracting, centrifuging, and transferring part of upper layer solution;

then adding 20 mu L of NaOH aqueous solution and 800 mu L of dichloromethane for secondary reverse extraction, oscillating and centrifuging;

and then adding 800 mu L of nitric acid solution into the lower layer solution for third extraction, oscillating and centrifuging to obtain an upper layer solution which is the solution to be detected.

Step 3, adding a ro-GO/AuNPs composite substrate into the solution to be detected, performing Raman scanning on the solution to be detected, and collecting spectral data in a corresponding range;

in the step, the specific process is as follows:

firstly, adding 600 mu L of ro-GO/AuNPs composite substrate and 80uL of nitric acid into 10 mu L of solution to be detected, and oscillating for 60 s;

then carrying out Raman scanning on the Raman-scanned optical fiber, wherein the specific parameters are as follows: the spectrum scanning time is 10s, the scanning frequency is 2, the smoothing parameter is 1, the scanning power is 100mw, and the collection is 1000-2000cm^-1Spectral data within a range.

And 4, carrying out △ v-1265, 1474 and 1602cm on the acquired spectral data^-1Raman absorption as characteristic qualitative peak of clenbuterol, and passing △ v-1474 cm^-1And quantitatively calculating the Raman absorption intensity to realize the detection of the clenbuterol.

Fig. 2 is a schematic diagram of characteristic qualitative peaks of clenbuterol provided in an embodiment of the present invention, where △ v 1265, 1474 and 1602cm are shown in fig. 2^-1The raman absorption was taken as the characteristic qualitative peak of clenbuterol.

FIG. 3 shows the present inventionThe clenbuterol quantitative curve provided by the example is shown in fig. 3 as △ v-1474 cm^-1And the Raman absorption peak is a clenbuterol quantitative peak for quantitative calculation.

By setting as described above, the detection sensitivity can be made to 1ng g^-1In the range of 1-100ng g^-1Quantitative calculation is realized within the range.

It is noted that those skilled in the art will recognize that embodiments of the present invention are not described in detail herein.

In conclusion, the detection method provided by the embodiment of the invention can greatly improve the stability of the composite SERS substrate (up to 240 days), has the advantages of rapidness, high sensitivity and good repeatability, and has the detection sensitivity of 1ng g^-1And the detection time is less than 10 min/sample, so that the detection efficiency is greatly improved, and the rapid screening of CL in animal-derived food is realized.

The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope of the present invention are included in the scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.