阅读说明：本技术 一种利用拉曼光谱快速检测白酒中氰化物的方法 (A method of quickly detecting cyanide in white wine using Raman spectrum ) 是由 夏婧竹 刘峰 盛金 殷磊 于 2018-05-25 设计创作，主要内容包括：本发明涉及一种利用拉曼光谱快速检测白酒中氰化物的方法,特别是适用于食品快速检测。具体为：以金纳米修饰的磁性微球溶胶作为拉曼检测增强剂,将1体积待测溶液和5体积拉曼检测增强剂进行混合,快速拉曼测定,根据特征峰位置和强度可实现氰化物的定性定量检测。本发明为白酒中的氰化物提供了一种新的快速检测的方法,该方法操作过程简便快速,结果准确,灵敏度高,检出限为0.01mg/L；此外配合手持拉曼分析仪可以实现现场和户外的实时分析。(The present invention relates to a kind of methods that cyanide in white wine is quickly detected using Raman spectrum, are particularly suitable for food science literature.Specifically: the magnetic microsphere colloidal sol modified using gold nano mixes 1 volume solution to be measured and 5 volume Raman detection reinforcing agents as Raman detection reinforcing agent, and fast Raman measurement can realize the qualitative and quantitative detection of cyanide according to characteristic peak positions and intensity.The present invention provides a kind of new method quickly detected for the cyanide in white wine, and this method operating process is easy quickly, and as a result accurately, high sensitivity, detection is limited to 0.01mg/L；This fit on, which holds Raman analyser, may be implemented scene and outdoor real-time analysis.)

1. a kind of method for quickly detecting cyanide in white wine using Raman spectrum, which is characterized in that be achieved by the steps of:

(1) by reduction of sodium citrate gold chloride, gold nanoparticle, uniform deposition to Fe are generated₃O₄Or Fe₂O₃Magnetic microsphere table Face, so that the complex microsphere (Au/Fe- complex microsphere) of surface covering Fe ion and nanometer Au is made, by Au/Fe- complex microsphere It is distributed in deionized water, obtains Raman detection reinforcing agent；

(2) negative electricity affecting agent is added into Raman detection reinforcing agent, makes the Zeta potential -60 of Au/Fe- complex microsphere colloidal sol ~-30 mV；

(3) a series of solution of cyanide standard items is prepared, or Wine Sample is handled；

(4) obtained Au/Fe- complex microsphere colloidal sol and 1 volume standard solution to be measured or to be measured will be handled in 5 debulking steps (2) Wine Sample is added sequentially in optical glass cuvette；

(5) under the conditions of 500 ~ 200 mW energy lasers and 2 ~ 10 second time of integration, with 2128 cm^-1±3 cm^-1Peak intensity is spy Sign draws intensity-concentration standard curve to various concentration cyanide standard solution, and in kind test Wine Sample is special Peak intensity is levied, sample to be tested feature peak intensity substitution standard curve is obtained into concentration of cyanide, to obtain to cyaniding in test sample The content of object.

2. a kind of method for quickly being detected cyanide in white wine using Raman spectrum according to claim 1, feature are existed In the Fe₃O₄Or Fe₂O₃The microballoon core of magnetic microsphere can be polystyrene or silica or other high molecular materials, micro- Ball surface uniform fold Fe₃O₄Or Fe₂O₃, Microsphere Size is 0.5 ~ 10 micron.

3. a kind of method for quickly being detected cyanide in white wine using Raman spectrum according to claim 1, feature are existed In the step (1), specially by 0.2gFe₃O₄Or Fe₂O₃Magnetic microsphere is added to 50mL and contains 0.1 ~ 0.2g HAuCl₄、3g NH₄In Cl, 2g sodium citrate mixed solution, it is centrifuged after ultrasonic reaction 5min.

4. a kind of method for quickly being detected cyanide in white wine using Raman spectrum according to claim 1, feature are existed In the Zeta potential is -50 ~ -40 mV.

5. a kind of method for quickly being detected cyanide in white wine using Raman spectrum according to claim 1, feature are existed In the negative electricity affecting agent can be one of sodium citrate, sodium ascorbate, tannic acid, or wherein the two or three Combination.

6. a kind of method for quickly being detected cyanide in white wine using Raman spectrum according to claim 1, feature are existed In the Wine Sample processing mode is that 5mL is taken to cross 0.22 μm of filter membrane, obtains prepare liquid.

7. a kind of method for quickly being detected cyanide in white wine using Raman spectrum according to claim 1, feature are existed In 2128 cm^-1±3 cm^-1If the characteristic peak that signal-to-noise ratio is greater than 3 occurs in position, show to contain cyaniding in sample to be tested Object.

Technical field

The invention belongs to the analysis detection fields of substance, especially technical field of rapid detection of food safety.

Background technique

Cyanide belongs to extremely toxic substance, into human body after cryanide ion is precipitated, with cell mitochondrial internal oxidition type cytochromes The ferric iron of oxidizing ferment combines, and prevents the ferric iron back in oxidizing ferment, interferes cell eupnea, causes histanoxia, lead Body is caused to fall into interior smother.In addition the molecule of certain nitrile compounds itself has the inhibition of direct Central nervous system Effect.The acute poisoning lethal dose of human oral's approach is 0.48 ~ 3.37 mg/kg weight (cyanogen root), is gone out in several minutes Existing acute poisoning symptom.

Cyanide is widely present in nature, and in plant, cyanide is usually in conjunction with glycan molecule, and with glucosides containing cyanogen (cyanogenic glycoside) form exists.For example, in cassava just contain glucosides containing cyanogen, using plant as raw material (such as cassava, Substitute, beans and other fruit stones are mixed into some wild plants) inevitably hydrolysis produces brewing white spirit during the fermentation Raw cyanide (such as HCN).Therefore, cyanide is an important monitoring index of drinks.GB2757-2012 is advised in national standard Being scheduled on cyanide limitation in Spirit and its assembled alcoholic drinks should be less than 8.0 g/L(in terms of HCN, convert by 100% alcoholic strength).

Currently, specifically including that optical colorimetry, spectrophotometry, gas phase color for the detection method of cyanide in food Spectrometry, the chromatography of ions and chromatography mass spectrometry etc..Wherein, optical colorimetry and spectrophotometry etc. mostly using cyanide and are shown Quantitative reaction between color substance, sensitivity is horizontal in 0.01 ~ 1 mg/ L, but specificity is poor；And the methods of chromatography (mass spectrum) Detection needs to perform the derivatization reaction or selects the chromatographic column of specific type, complex disposal process, time-consuming, at high cost, needs specially Industry technical operation, and it is not suitable for field quick detection.The present invention develops a kind of using Raman spectrum, quickly analyzes in white wine The method of cyanide.Raman spectrum nineteen twenty-eight is found, and nineteen thirty obtains Nobel Prize in physics, it is prevalent in a cutting In son, the structural information of molecule can be reliably provided, do not influenced by aqueous solvent etc., with the use of laser light source, laser is drawn Graceful spectrum has become important compound analysis means, is widely used in the fields such as criminal investigation identification, mineral analysis, in recent years With surface enhancement techniques, the combination of Fourier's infrared technique and laser Raman spectroscopy, start to be applied to food safety detection.

Summary of the invention

In order to solve the above-mentioned technical problem, the object of the present invention is to provide a kind of Ramans of cyanide in quickly detection white wine Enhanced spectrum method, to realize fast qualitative and the quantitative analysis to cyanide in white wine.

The present invention is achieved through the following technical solutions.

A method of quickly detecting cyanide in white wine using Raman spectrum, this method comprises the following steps:

(1) by reduction of sodium citrate gold chloride, gold nanoparticle, uniform deposition to Fe are generated₃O₄Or Fe₂O₃Magnetic microsphere table Face, so that the complex microsphere (Au/Fe- complex microsphere) of surface covering Fe ion and nanometer Au is made, by Au/Fe- complex microsphere It is distributed in deionized water, obtains Raman detection reinforcing agent；

(2) negative electricity affecting agent is added into Raman detection reinforcing agent, makes the Zeta potential -60 of Au/Fe- complex microsphere colloidal sol ~-30 mV；

(3) a series of solution of cyanide standard items is prepared, or Wine Sample is handled；

(4) obtained Au/Fe- complex microsphere colloidal sol and 1 volume standard solution to be measured or to be measured will be handled in 5 debulking steps (2) Wine Sample is added sequentially in optical glass cuvette；

(5) under the conditions of 500 ~ 200 mW energy lasers and 2 ~ 10 second time of integration, with 2128 cm^-1±3 cm^-1Peak intensity is Feature draws intensity-concentration standard curve to various concentration cyanide standard solution, and in kind tests Wine Sample Sample to be tested feature peak intensity substitution standard curve is obtained concentration of cyanide, to obtain to cyanogen in test sample by feature peak intensity The content of compound.

Wherein, Fe described in step (1)₃O₄Or Fe₂O₃Magnetic microsphere, microballoon core can be polystyrene or titanium dioxide Silicon or other high molecular materials, microsphere surface uniform fold Fe₃O₄Or Fe₂O₃, Microsphere Size is 0.5 ~ 10 micron.

Specific preparation method described in step (1) is by 0.2gFe₃O₄Or Fe₂O₃Magnetic microsphere is added to 50mL and contains 0.1 ~0.2g HAuCl₄、3g NH₄In Cl, 2g sodium citrate mixed solution, it is centrifuged after ultrasonic reaction 5min.

Zeta potential described in step (2), preferably -50 ~ -40 mV, especially -45 mV.

Negative electricity affecting agent described in step (2) can be one of sodium citrate, sodium ascorbate, tannic acid, or The wherein combination of the two or three.

Wine Sample processing mode described in step (3) is that 5 mL is taken to cross 0.22 μm of filter membrane, obtains prepare liquid.

It may be noted that observing 2128 cm in above-mentioned steps (5)^-1±3 cm^-1If there is the spy that signal-to-noise ratio is greater than 3 in position Peak is levied, if showing to contain cyanide in sample to be tested, and then is quantitative determined.

The invention has the advantages that providing a kind of Raman enhanced spectrum method for quickly detecting cyanide in white wine, lead to It crosses and prepares a kind of Au/Fe- complex microsphere colloidal sol, under suitable Zeta potential, can be combined very well with cyanide, generate Raman Enhance enlarge-effect, realizes the trace detection analysis of cyanide in white wine.The present invention in test, is an attempt to preparation nanometer originally The microballoon of gold package is Raman enhancing agent, Au and cyanogen root can be avoided to be coordinated to form solubility by the stabilization core-shell structure of microballoon Compound Au (CN)^2-Stable Raman detection signal is obtained, but obtained signal strength is very weak.Later using in magnetic microsphere Surface modification on nanogold particle, the Au/Fe- complex microsphere being prepared obtains significant Raman enhancement effect, this is answered This is the Fe of magnetic microsphere surface package₃O₄Or Fe₂O₃Ferric ion can be produced, is had strongly to the cyanogen root in sample Combination, by nanogold and ferric collective effect, produces to realize the rapid efficiently concentrating of trace cyanogen root in sample Raw strong Raman signal.Sample substrate background has little effect detection in the invention, without special sample pre-treatments, It only needs simply to be filtered, can realize in several minutes and the rapid sensitive of cyanide in white wine is detected, the detection method Fast, sensitive, operation profession require it is low, be particularly suitable for cooperation Handheld Raman analyzer realize scene and it is outdoor in real time point Analysis.

Detailed description of the invention

Fig. 1 is the surface-enhanced Raman map of 5 mg/L cyanide standard solution.

Fig. 2 is Wine Sample and is added to the surface-enhanced Raman map of 0.3mg/L cyanide standard specimen in Wine Sample.

Specific embodiment

Below in conjunction with attached drawing 1, attached drawing 2, the present invention is described in further detail.

(1) synthesis of Au/Fe- complex microsphere: by 0.2gFe₃O₄Magnetic microsphere is added to 50mL and contains 0.2g HAuCl₄、 3g NH₄In Cl, 2g sodium citrate mixed solution, it is centrifuged after ultrasonic reaction 5min.

(2) adjusting of Raman detection reinforcing agent: Au/Fe- complex microsphere is distributed in deionized water, and it is multiple to adjust Au/Fe- The Zeta potential for closing microballoon colloidal sol is -50 mV, obtains Raman enhancing agent.

(3) it configures cyanide standard solution: accurately weighing 0.1 g cyanide standard items, after being dissolved with water, constant volume to 100 ML, 4 DEG C are kept in dark place, and standard solution is diluted to 0.1mg/L, 0.5mg/L, 1mg/L, 2 mg/L, 5mg/L, 10mg/L respectively Series standard solution for standby.

(4) it Wine Sample solution preparation to be measured: takes 5 mL white wine to cross 0.22 μm of filter membrane, obtains prepare liquid.

(5) standard curve and test sample are drawn: setting wavelength the swashing as 785 nm lasers of Raman spectrometer institute band 300 mW of optical power, 2 s of the time of integration, average time 2 times, smoothing parameter 5,300 ~ 3500cm of scanning range^-1.For each mark 5 volume Au/Fe- complex microsphere colloidal sol Raman detection reinforcing agents, 1 volume are added in quasi- solution in 1 cm optical glass cuvette Standard sample, place it in laser Raman spectrometer and detect indoor detection.With 2128cm^-1± 3cm^-1Peak intensity is characterized, Various concentration cyanide standard solution is measured by this condition respectively, draw intensity-concentration standard curve, and test to Survey liquid feature peak intensity.

In the present embodiment, with concentration of cyanide (mg/L) for abscissa, peak intensity is ordinate, draws standard curve, The result shows that linear relationship is good in the range of 0.1 mg/L to 10mg/L, regression coefficient R²=0.998, Fig. 1 are 5 mg/L The Raman map of standard solution, it is seen that in 2128 cm^-1There are characteristic Raman signals strongly.

Under identical testing conditions, certain Wine Sample is detected, as a result as shown in Figure 2, it can be seen that in wavelength 2128 cm^-1There are apparent Raman signals at place.

To further determine whether the characteristic peak for cyanide, the cyanide standard of 0.3mg/L is added into Wine Sample Solution finds that Raman signal is remarkably reinforced, therefore in this characteristic peak positions, it is possible to determine that, cyanide is contained in the white wine, and This signal strength is calculated on cyanide standard curve, the content of cyanide is 0.08 mg/L in Wine Sample.

For the repeatability for verifying the detection method, Wine Sample is subjected to detection 5 times in different time, the results are shown in Table 1:

Cyanide repeatability testing result in 1 Wine Sample of table

Pendulous frequency	1	2	3	4	5	Relative standard deviation RSD (%)
							Cyanide content (mg/L)	0.078	0.075	0.082	0.076	0.084	4.38

As it can be seen from table 1 in 5 white wine of METHOD FOR CONTINUOUS DETERMINATION cyanide content, RSD 4.38%, illustrate the Raman spectroscopy survey It is higher to determine cyanide repeatability.

In the present embodiment, to the mark in Wine Sample, being separately added into 0.1 mg/L, 0.2mg/L, 0.3mg/L, 0.4mg/L Quasi- solution is detected after filtration treatment using above-mentioned Raman spectroscopy, is calculated recovery of standard addition, be the results are shown in Table 2:

The recovery of standard addition result of cyanide in 2 Wine Sample of table

Scalar quantity (mg/L)	The rate of recovery (%)
		0.1	82
0.2	85
		0.3	89
0.4	88

From table 2 it can be seen that the recovery of standard addition of cyanide illustrates that the detection method is returned between 82% ~ 89% in Wine Sample Yield is higher.

Further, it is obtained by calculation, under the premise of signal-to-noise ratio S/N=3, cyaniding in detection white wine provided by the invention The method of object, detection limit is up to 0.01mg/L.

Above embodiments illustrate application method of the invention, but the content that the present invention is protected is not limited only to embodiment In usage, the improvement for the category nature extended nature that professional and technical personnel in the field carry out according to the present invention also belongs to of the invention Protection scope.