CN113866100B - Method for judging authenticity of white skill by utilizing nano silver deposition visual sensor - Google Patents
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- 230000008021 deposition Effects 0.000 title claims abstract description 18
- 230000000007 visual effect Effects 0.000 title claims abstract description 13
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 title claims abstract description 11
- 241000132012 Atractylodes Species 0.000 claims abstract description 46
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- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims abstract description 27
- 229910052709 silver Inorganic materials 0.000 claims abstract description 12
- 239000004332 silver Substances 0.000 claims abstract description 12
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- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 9
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- 230000008859 change Effects 0.000 claims description 5
- 238000012569 chemometric method Methods 0.000 claims description 5
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 4
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- 238000004458 analytical method Methods 0.000 description 4
- WOWHHFRSBJGXCM-UHFFFAOYSA-M cetyltrimethylammonium chloride Chemical compound [Cl-].CCCCCCCCCCCCCCCC[N+](C)(C)C WOWHHFRSBJGXCM-UHFFFAOYSA-M 0.000 description 4
- 238000012549 training Methods 0.000 description 4
- LZZYPRNAOMGNLH-UHFFFAOYSA-M Cetrimonium bromide Chemical compound [Br-].CCCCCCCCCCCCCCCC[N+](C)(C)C LZZYPRNAOMGNLH-UHFFFAOYSA-M 0.000 description 3
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- SQGYOTSLMSWVJD-UHFFFAOYSA-N silver(1+) nitrate Chemical compound [Ag+].[O-]N(=O)=O SQGYOTSLMSWVJD-UHFFFAOYSA-N 0.000 description 2
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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
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
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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
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/29—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands using visual detection
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Abstract
The invention discloses a method for judging authenticity of white atractylodes rhizome by utilizing a nano silver deposition visual sensor, which comprises the steps of firstly uniformly mixing white atractylodes rhizome or rhizoma atractylodis extracting solution with nano gold, then respectively introducing silver ions with the same concentration, carrying out heating reaction, and establishing a relation between a white atractylodes rhizome or rhizoma atractylodis sample and the color difference according to deposition differences of silver ions on the surface of the nano gold in a reaction system obtained by utilizing different white atractylodes rhizome or rhizoma atractylodis extracting solutions and reaction system color differences caused by the deposition differences, thereby realizing authenticity judgment of unknown white atractylodes rhizome products. The detection method has the advantages of low cost, quick response, visualization, convenient operation, no need of large-scale precise instruments and the like, can rapidly realize the precise judgment of the authenticity of the bighead atractylodes rhizome, can effectively solve the problems of high cost, complex operation, high requirement, long detection time and the like of the existing detection method, and has important research, popularization and application.
Description
Technical Field
The invention belongs to the technical field of chemical analysis and detection, and particularly relates to a method for judging authenticity of a white skill by using a nano silver deposition visual sensor.
Background
The Atractylodis rhizoma is perennial herb of Atractylodes of Compositae, and its dried rhizome is a common tonic Chinese medicinal material, and has effects of invigorating spleen, invigorating qi, eliminating dampness, promoting diuresis, suppressing sweating, and preventing miscarriage. The current research shows that the extract mainly contains volatile oil (mainly atractylone and atractylenolide), polysaccharide and a small amount of terpenoid compounds, has various functional activities such as anti-tumor, antibacterial, anti-inflammatory and antioxidant activities, and has wide application prospect. However, due to the numerous production places, the quality of the bighead atractylodes rhizome of different varieties is uneven. In addition, the Chinese atractylodes belonging to the same genus is similar to the Chinese atractylodes in terms of its property, ingredient and efficacy, and both are easy to be confused. Therefore, the existing authenticity detection problem of the bighead atractylodes rhizome is further explored and optimized, and the method has important research and application significance.
The existing method for judging the authenticity of the bighead atractylodes rhizome at home and abroad mainly comprises a high-performance liquid chromatography, a high-performance liquid chromatography-mass spectrometry technology, a gas chromatography-mass spectrometry technology, a high-performance capillary electrophoresis method and the like, and the analysis methods have the defects of high precision, strong anti-interference capability, accurate qualitative and quantitative determination, but have the defects of expensive instruments, long analysis time, complicated maintenance, large-scale instruments requiring professional operators and the like, and are not beneficial to popularization. It is necessary to further establish a rapid, simple and low-cost detection method for judging the authenticity of the bighead atractylodes rhizome.
Disclosure of Invention
Aiming at the defects existing in the prior art, the invention provides the method for detecting the authenticity of the bighead atractylodes rhizome based on the nano-gold surface silver deposition visual sensor, which has the advantages of low cost, quick response, visualization, convenient operation and the like, can rapidly realize the accurate judgment of the authenticity of the bighead atractylodes rhizome, and is suitable for popularization and application.
In order to achieve the above purpose, the invention adopts the following technical scheme:
A method for judging authenticity of a white skill by utilizing a nano silver deposition visual sensor comprises the following steps: respectively extracting a white atractylodes rhizome sample and a rhizoma atractylodis sample, uniformly mixing the obtained white atractylodes rhizome or rhizoma atractylodis extracting solution with nano gold, respectively introducing silver ions with the same concentration, performing heating reaction, and judging the authenticity of an unknown white atractylodes rhizome product according to the deposition difference of the silver ions on the surface of the nano gold and the color difference of a reaction system caused by the deposition difference of the silver ions in a reaction system obtained by using different white atractylodes rhizome or rhizoma atractylodis extracting solutions.
In the scheme, the distinguishing method comprises the steps of directly and visually observing the color difference to distinguish the bighead atractylodes rhizome from the rhizoma atractylodis or establishing the relationship between the bighead atractylodes rhizome or rhizoma atractylodis sample and the color difference by adopting a chemometric method.
In the scheme, the extracting agent adopted in the extracting step is methanol solution or ethanol solution with the concentration of 45-55wt%; the solid-liquid ratio between the extractant and the atractylodes macrocephala koidz or atractylodes macrocephala koidz sample is 1g (300-500) mL.
In the scheme, the extraction process adopts conventional extraction processes such as ultrasonic extraction and the like.
In the above scheme, the Ag + is introduced by adopting inorganic silver salts such as silver nitrate.
In the above scheme, the concentration of Ag + in the reaction system is (1-3). Times.10. 10 -4 mol/L.
In the scheme, the rhizoma atractylodis extracting solution accounts for 35-45% of the volume of the reaction system.
In the scheme, the molar ratio of the nano gold to the Ag + in the reaction system is 1 (10 4～1.2×105).
In the scheme, the heating reaction temperature is 50-70 ℃ and the time is 10-25min.
In the scheme, the color information is obtained by converting the color change of the nanogold before and after the reaction of the introduced atractylodes macrocephala koidz extract or the atractylis ovata extract into an RGB value; and adopting chemometric method to construct the relationship between RGB values and rhizoma Atractylodis Macrocephalae or rhizoma Atractylodis products in different places.
In the above-described embodiment, the chemometric method may be Partial Least Squares Discriminant Analysis (PLSDA), or the like.
In the scheme, the average grain diameter of the nano gold is 12-14 nm, and the morphology of the nano gold is spherical.
In the above scheme, the preparation method of the nano gold comprises the following steps:
1) Preparing a mixed aqueous solution (yellow solution) of chloroauric acid and a surfactant CTAB, then adding a reducing agent sodium borohydride under stirring, and carrying out stirring reaction to obtain an initial nano gold seed solution (light pink);
2) Uniformly mixing chloroauric acid and CTAC, preparing a mixed aqueous solution of chloroauric acid and CTAC, adding ascorbic acid, then adding the nano gold seed solution obtained in the step 1) under the condition of rapid stirring, and reacting to obtain the nano gold dispersion (pink).
In the scheme, the molar ratio of chloroauric acid to CTAB to sodium borohydride in the step 1) is 1 (350-450): 4-5.
In the scheme, the molar ratio of chloroauric acid to CTAC to ascorbic acid in the step 2) is 1 (0.3-0.5) (130-180).
In the scheme, the molar ratio of the chloroauric acid adopted in the step 1) to the chloroauric acid adopted in the step 2) is 1 (75-85).
In the scheme, the stirring reaction time in the step 1) is 2-4h; the reaction time in the step 2) is 5-15.
The principle of the invention is as follows:
The invention is based on the characteristic that Ag + can be deposited on the surface of nano gold (Ag + deposition of different degrees shows the color change from powder to orange), and firstly proposes to utilize the characteristic that the effective components in the extracting solution of bighead atractylodes rhizome and rhizoma atractylodis are used for preventing Ag + from being deposited on the surface of nano gold, according to the different relations of silver deposition preventing capability caused by different contents of effective compounds in bighead atractylodes rhizome and rhizoma atractylodis in different places, the RGB value of the surface of nano gold in a colorimetric plate after silver deposition is extracted by PS software, and color information is further processed by combining a Partial Least Squares Discriminant Analysis (PLSDA) model, so that the sensor for visually judging the authenticity of bighead atractylodes rhizome is constructed.
Compared with the prior art, the invention has the beneficial effects that:
The invention provides a rapid visual detection method for the authenticity of the bighead atractylodes rhizome for the first time by utilizing the characteristic that the effective components in the bighead atractylodes rhizome and rhizoma atractylodis extract prevent Ag + from depositing on the surface of nano gold and having difference; the detection method has the advantages of low cost, quick response, visualization, convenient operation, no need of large-scale precise instruments and the like, can rapidly realize the precise judgment of the authenticity of the bighead atractylodes rhizome, can effectively solve the problems of high cost, complex operation, high requirement, long detection time and the like of the existing detection method, and has important research, popularization and application.
Drawings
Fig. 1 is a schematic flow chart of a method for judging the authenticity of bighead atractylodes rhizome by using the visual sensor based on nano-gold surface silver deposition.
FIG. 2 is a TEM characterization of CTAC-AuNPs nanogold obtained in example 1.
FIG. 3 shows the visual color reaction of three kinds of bighead atractylodes rhizome and rhizoma atractylodis in origin detected by using nano gold in example 1.
FIG. 4 shows the result of partial least squares discriminant analysis of three kinds of bighead atractylodes rhizome and rhizoma atractylodis in example 1.
Detailed Description
The present invention will be described in further detail with reference to the following examples in order to make the objects, technical solutions and advantages of the present invention more apparent. It should be understood that the specific embodiments described herein are for purposes of illustration only and are not intended to limit the scope of the invention.
Example 1
A method for judging authenticity of a white skill by utilizing a nano silver deposition visual sensor comprises the following steps:
1) Preparation of nano gold: adding 5ml of HAuCl 4 aqueous solution (0.25 mM) and 5ml of CTAB aqueous solution (100 mM) into a beaker to obtain a yellow solution, adding 600ul of freshly prepared NaBH 4 aqueous solution (10 mM) under magnetic stirring, immediately turning the color of the obtained solution system from yellow to colorless, and continuously stirring for 3 hours to change the color into light pink to obtain an initial nano gold seed solution; adding 6ml of HAuCl 4 aqueous solution (0.5 mM) and 6ml of CTAC aqueous solution (0.2M) into another clean beaker, uniformly mixing, adding 4.5ml of AA aqueous solution (100 mM), then adding 300ul of the nano gold seed solution obtained in the above step under the condition of rapid stirring, and changing the color of a solution system from colorless to pink to obtain CTAC-AuNPs nano gold solution (the concentration is 3.4 nmol/L);
2) Taking 6 atractylodes macrocephala samples of Zhejiang, hunan and Anhui province, and 18 atractylodes samples of different producing areas (including two samples of inner Mongolian Tongliao, two samples of inner Mongolian producing areas, four samples of inner Mongolian wild, and seven samples of Anhui Bozhou; two batches of Hubei yellow spine samples; a batch of Hebei Anguo samples), crushing the block-shaped samples, sieving the crushed block-shaped samples with a 100-mesh sieve, and respectively carrying out ultrasonic extraction on the powder samples, wherein the specific process conditions comprise: extracting with CNC ultrasonic cleaner (KQ-500 DE) with 50wt% methanol solution at a feed-liquid ratio of 1g:400mL at room temperature for 30min, and filtering to obtain Atractylodis rhizoma extractive solution and rhizoma Atractylodis extractive solution;
3) Taking 500 mu L (3.4 nmol/L) of CTAC-AuNP nano gold solution, adding 500 mu L of water, uniformly mixing, and taking the solution as a blank solution; taking 500 mu L of CTAC-AuNP nano-gold solution (3.4 nmol/L), 400 mu L of water and 100 mu L of AgNO 3 solution with concentration of 2X 10 - 3 mol/L, uniformly mixing, and taking the mixture as a control group; taking 500 mu L of CTAC-AuNP nano gold solution (3.4 nmol/L), 400 mu L of white atractylodes rhizome or rhizoma atractylodis extract, uniformly mixing, and then adding 100 mu L of AgNO 3 solution (2X 10 -3 mol/L) into the mixture to serve as an experimental group; wherein each blank group, control group or experimental group controls the reaction time to be 20min, and the reaction temperature is 60 ℃;
3) Using a camera to obtain pictures of the nano gold in the 96-well plate after the reaction of different reaction systems, and obtaining color information in the pictures; extracting 10×10 pixel points by using Photoshop 14.0 software, and converting the picture information into RGB numerical information by using Matlab R2016 a;
4) Performing model discriminant analysis (the division of training set and prediction set is shown in table 1) by using Partial Least Squares Discriminant Analysis (PLSDA), preparing 18 samples of rhizoma Atractylodis in each producing area, and preparing 18 samples of rhizoma Atractylodis in different producing areas; and establishing the relationship between the RGB numerical information and different products of bighead atractylodes rhizome and rhizoma atractylodis.
Fig. 2 is a TEM characterization diagram of CTAC-AuNPs nanogold obtained in this example, and it can be seen that the nanogold presents spherical nanoparticles, and the particle size is mainly about 12 nm.
Fig. 3 shows the results of the visual color reaction test of three kinds of producing areas of bighead atractylodes rhizome and rhizoma atractylodis by adopting nano gold in the embodiment, the color changes of RGB of the bighead atractylodes rhizome and the rhizoma atractylodis are obviously different, the results obtained by the bighead atractylodes rhizome sample are red brown, and the results obtained by the rhizoma atractylodis sample are lighter in color and mainly light purple and pale yellow. The comparison of the reaction results of the bighead atractylodes rhizome in different producing areas shows that the reaction results of the three bighead atractylodes rhizome have similar colors and have smaller RGB color change difference; based on the visual result, the bighead atractylodes rhizome and the rhizoma atractylodis can be directly distinguished under naked eyes.
Table 1 division of training set and prediction set for plausibility PLS-DA mode discrimination of Atractylodis rhizoma (state=5)
TABLE 2 PLS-DA discrimination results of the authenticity of Atractylodis rhizoma
As shown in fig. 4 and table 2, the specific discrimination results according to the above scheme are that the number of training set samples is larger than the number of prediction set samples. There are three bighead atractylodes rhizome producing areas and one rhizoma atractylodis, and 18 samples are taken under each category. When state=5, selecting the optimal hidden variable as 5, wherein under the condition, the accuracy of the training set is 98.65%, and the accuracy of the prediction set is 97.06%; the method can rapidly realize accurate judgment of the authenticity of the bighead atractylodes rhizome.
The above examples are presented for clarity of illustration only and are not limiting of the embodiments. Other variations and modifications of the above description will be apparent to those of ordinary skill in the art, and it is not necessary or exhaustive of all embodiments, and thus all obvious variations or modifications that come within the scope of the invention are desired to be protected.
Claims (4)
1. The method for judging the authenticity of the white skill by using the nano silver deposition visual sensor is characterized by comprising the following steps of: respectively extracting a white atractylodes rhizome sample and a rhizoma atractylodis sample, uniformly mixing the obtained white atractylodes rhizome or rhizoma atractylodis extracting solution with CTAC-AuNPs dispersion liquid, respectively introducing silver ions with the same concentration, performing heating reaction, and judging the authenticity of an unknown white atractylodes rhizome product according to the deposition difference of the silver ions on the surface of nano gold and the color difference caused by the deposition difference of the silver ions in a reaction system obtained by using different white atractylodes rhizome or rhizoma atractylodis extracting solutions; the distinguishing method comprises directly observing color difference with naked eyes to distinguish Atractylodis rhizoma and rhizoma Atractylodis or adopting chemometric method to establish relationship between Atractylodis rhizoma or rhizoma Atractylodis sample and color difference;
The shape of the nano gold is spherical;
the solid-to-liquid ratio between the extractant and the atractylodes macrocephala koidz or rhizoma atractylodis sample is 1g (300-500) mL;
In the reaction system, the concentration of Ag + is (1-3) multiplied by 10 -4 mol/L, and the molar ratio of nano gold to Ag + is 1 (10 4～1.2×105);
The volume percentage of the extracting solution in the reaction system is 35-45%;
the heating reaction temperature is 50-70 ℃ and the time is 10-25min.
2. The method according to claim 1, wherein the extracting agent used in the extracting step is a methanol solution or an ethanol solution having a concentration of 45-55 wt%.
3. The method according to claim 1, wherein the color information is obtained by converting color change of nanogold before and after the reaction of introducing the Atractylodis rhizoma extract or the rhizoma Atractylodis extract into RGB values; and then adopting a chemometric method to construct the relationship between RGB values and white atractylodes rhizome or rhizoma atractylodis products in different places.
4. The method according to claim 1, wherein the average particle size of the nanogold is 12-14 nm.
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