CN106706594A - Method for achieving simultaneous detection of chemical structure and biological activity of functional component of food - Google Patents
Method for achieving simultaneous detection of chemical structure and biological activity of functional component of food Download PDFInfo
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- G—PHYSICS
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- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- 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
- G01N21/65—Raman scattering
- G01N21/658—Raman scattering enhancement Raman, e.g. surface plasmons
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
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- 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
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- 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
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
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Abstract
The invention discloses a method for achieving simultaneous detection of a chemical structure and biological activity of a functional component of a food, and relates to a method for representing an active component in the food. The method comprises the steps of carrying out surface-enhanced raman scattering on a to-be-tested sample, carrying out second-order conversion and peak-fit processing on a raman scattering map to obtain a second-order conversion map and determining characteristic peaks of the active component; carrying out peak intensity value accumulation on the areas which are +/-5cm<-1> from valley positions of the characteristic peaks of the active component to obtain peak intensity accumulation values; carrying out normalization processing on the peak intensity accumulation values by adopting the maximum peak intensity accumulation value as a reference peak, and calculating bar code band width values of all characteristic peaks; and drawing a characteristic bar code of the active component according to the distribution positions of the characteristic peaks in the second-order conversion map and the bar code band width values. A technique for representing the active component in the food by using the bar code is provided. The biological activity of the active component is associated with chemical structure characteristics, so that simultaneous recognition of the structure and the activity is achieved. A simple and efficient active component detection method is provided.
Description
Technical field
This disclosure relates to technical field of food detection, in particular it relates to one kind realize food functional component chemical constitution and
The method that bioactivity is detected simultaneously.
Background technology
Such as phenols, isoprenoid, polyunsaturated fatty acid, protein, amino acid and the carbon aquation being rich in food
The food such as derivative of compound functional component has regulation immunity, improves blood pressure, blood fat, blood sugar level, builds up health, and prevents
The important function such as cardiovascular and cerebrovascular disease.But due to the complexity and diversity of food substrate, to the detection of these functional components
Cause difficulty.
SERS (Surface-enhanced Raman Scattering, SERS) has specimen preprocessing
Reason is simple, the detection used time is short, detect the advantages such as consumption is few, test limit is low, detection method is easy, testing result is accurate, especially
Portable hand-held Raman detection equips the Site Detection of achievement unit go-on-go test sample product, monitoring food has been successfully applied at present and has been lived
Property constituent structure change procedure, while also studies have reported that it is in isoprenoid derivative, aldehydes matter, with protein or ammonia
The Active components of food Structural Identifications such as derivative, carbohydrate and its derivative and polyunsaturated fatty acid based on base acid
Aspect has a clear superiority, while " finger-print " characteristic that SERS technologies have is further to realize structure with spectrogram just
Victory conversion provides the foundation.But SERS is not a kind of achievable functional component chemical constitution and bioactivity identification simultaneously
Technology, exclusive use cannot generally illustrate the bioactivity of complex biological activity functional component, therefore SERS bar codes techniques institute
The collection bioactivity that has and the outstanding advantage of chemical structure information can be used to realizing to complex biological activity effect in food into
Recognized while the chemical constitution divided is with bioactivity.
The content of the invention
The purpose of the disclosure is to provide a kind of chemical constitution to bioactive ingredients in food and carries out letter with bioactivity
Just the method with qualitative recognition is efficiently characterized.
To achieve the above object, the one side of the disclosure provides a kind of method for characterizing Active Ingredient in Food, bag
Include following steps：
SERS is carried out to testing sample, second order conversion is carried out to Raman scattering collection of illustrative plates and peak-fit processing is obtained
Obtain second order conversion spectrogram and determine the characteristic peak of active component, to the characteristic peak wave trough position ± 5cm of the active component-1Area
Domain carries out the cumulative peak intensity that obtains of peak intensity value and adds and be worth, and peak intensity is added and is worth as reference peaks using peak intensity plus with the maximum characteristic peak of value
It is normalized, the bar coded strip width value of reference peaks is set to special value X, is calculated a's using formula I
Value, the bar coded strip width value of all characteristic peaks is calculated using a values, according to second order change spectrogram in each characteristic peak divide
Cloth position and the bar coded strip width value draw the feature bar code of the active component, in the feature bar code,
Each bar coded strip characterizes a chemical structure information of the active component；
Formula I：Bar coded strip width value=a × peak intensity adds and is worth.
Optionally, methods described also includes, will characterize the particular organisms activity related chemical structure of the active component
Bar coded strip is defined as the activity band of the bioactivity, and activity band color 1 is marked, will except the activity band with
Other outer bar coded strips are marked with other colors for having cognizable difference with color 1.
Optionally, the span of a is 1/10000-1/1000, and the special value X is 10 pounds.
Optionally, the testing sample is the standard items of active component, sets up the feature bar code data of the standard items
Storehouse.
Optionally, the condition of the SERS include laser wavelength be 325nm, 514nm, 532nm,
633nm or 785nm, sweep time is 10-30s, and scanning times are 2-5 times, and wave-number range is 0-4000cm-1。
Optionally, the substrate used by SERS is silver-colored dendritic nano particle.
Optionally, the preparation method of silver-colored dendritic nano particle includes：It is 100-200mmol/L's that zinc metal sheet is put into concentration
AgNO320-60s is reacted in solution.
Optionally, the active component is including isoprenoid derivative, aldehydes matter, with protein or amino acid as base
The derivative of plinth, carbohydrate and its derivative and polyunsaturated fatty acid.
Second aspect of the disclosure provides a kind of method for detecting Active Ingredient in Food, comprises the following steps：
S1：The feature bar code B of active component in testing sample is obtained using the method described in disclosure one side；
S2：The feature bar code B is compared with the bioactivity of each active component standard items, by testing sample
The chemical constitution of active component be labeled to realize Qualitive test in bar code.
Optionally, it is characterised in that methods described also includes detecting the bioactivity of active component standard items, incites somebody to action
Bioactivity testing result and Structure-activity analysis result are compared with the feature bar code of the standard items, obtain feature bar
Relation between the chemical structure information and bioactivity of the active component reflected in shape code.
The disclosure is provided a kind of using bar code sign Active Ingredient in Food chemical constitution by above-mentioned technical proposal
Technology, while the bioactivity of active component can also be associated with its chemical structure characteristic, so as to realize structure with
Characterized while active and detected, there is provided a kind of active component detection method of simple and effective.
Other feature and advantage of the disclosure will be described in detail in subsequent specific embodiment part.
Brief description of the drawings
Accompanying drawing is, for providing further understanding of the disclosure, and to constitute the part of specification, with following tool
Body implementation method is used to explain the disclosure together, but does not constitute limitation of this disclosure.In the accompanying drawings：
Fig. 1 is 9 kinds of chemical constitution schematic diagrames of active component；
Fig. 2 is 9 kinds of second order SERS spectrograms of active component；
Fig. 3 is 9 kinds of bioactivity testing results of active component；
Fig. 4 is 9 kinds of feature bar codes of active component.
Specific embodiment
It is described in detail below in conjunction with accompanying drawing specific embodiment of this disclosure.It should be appreciated that this place is retouched
The specific embodiment stated is merely to illustrate and explains the disclosure, is not limited to the disclosure.
The one side of the disclosure provides a kind of method for characterizing Active Ingredient in Food chemical constitution, including following step
Suddenly：
SERS is carried out to testing sample, second order conversion is carried out to Raman scattering collection of illustrative plates and peak-fit processing is obtained
Obtain second order conversion spectrogram and determine the characteristic peak of active component, to the characteristic peak wave trough position ± 5cm of the active component-1Area
Domain carries out the cumulative peak intensity that obtains of peak intensity value and adds and be worth, and peak intensity is added and is worth as reference peaks using peak intensity plus with the maximum characteristic peak of value
It is normalized, the bar coded strip width value of reference peaks is set to special value X, is calculated a's using formula I
Value, the bar coded strip width value of all characteristic peaks is calculated using a values, according to second order change spectrogram in each characteristic peak divide
Cloth position and the bar coded strip width value draw the feature bar code of the active component, in the feature bar code,
Each bar coded strip characterizes a chemical structure information of the active component；
Formula I：Bar coded strip width value=a × peak intensity adds and is worth.
Wherein, the position one that the position of each band occurs with each characteristic peak in second order conversion collection of illustrative plates in the feature bar code
One correspondence.
Be imparted to for bioactivity information according to biological activity test result and structure-activity relationship by the method that the disclosure is provided
In bar code, feature bar code, the relation between chemical constitution and bioactivity are set up.The feature bar code of the active component
Intrinsic characteristic comprising structure-activity relationship, convenient identification while being capable of achieving structure with bioactivity.
Wherein, the multiplication coefficient that the coefficient a adds and be worth for peak intensity, specifically, if the peak of certain active component reference peaks
It is N to force and be worth, and its bar coded strip width value such as is set into special value X pounds, then according to " bar code width value=a
× peak intensity adds and is worth " calculate it is known that a=X/N, after a values are calculated, it is possible to use a values calculating active component its
The bar coded strip width value of its characteristic peak；In one kind of the disclosure compares preferred embodiment, for the ease of calculating, can
An integer value for being easy to calculate is set to by reference peaks bar coded strip width value, for example, can set it to 10 pounds, a
Span be 1/10000-1/1000.
Optionally, the chemical constitution feature of the active component in the testing sample can be labeled in bar code
To realize Qualitive test, methods described also includes, will characterize the particular organisms activity related chemical structure of the active component
Bar coded strip is defined as the activity band of the bioactivity, and activity band color 1 is marked, will except the activity band with
Other outer bar coded strips are marked with other colors for having cognizable difference with color 1.
In a kind of specific embodiment of the disclosure, a certain bioactivity of active component is anti-oxidant work as described
Property, because the feature bar code can characterize its chemical structure information, then those skilled in the art can be from its feature bar
The chemical structure information related to its antioxidation activity is read in shape code, characterize band as activity of the chemical constitution into
Divide the activity band of antioxidation activity, the band color 1 can be marked, remaining band is had and can recognized with color 1
Difference other colors mark.The color of the feature bar code should have cognizable difference with background when drawing bar code
It is different.Such as background color can be white, and color 1 can be black, and other colors for having cognizable difference with color 1 can
Think grey.
In the method that the disclosure is provided, described to have cognizable difference refer to that the difference of the two can be by this area
Technical staff can make a distinction by naked eyes or readily discriminating by conventional instrument.
Preferably, methods described includes screening spectral region, crosses noise filtering, improves signal to noise ratio so as to obtain feature
Peak, in the method that the disclosure is provided using second order conversion abate the noise and peak-fit processing after obtain second order conversion spectrogram, it is determined that
The characteristic peak of active component in second order collection of illustrative plates.Specifically, can be carried out by OMNIC and TQ Analyst data processing softwares
The second order conversion of SERS spectrograms, to system immanent structure relation dimensionality reduction, eliminates change at random.
Wherein, the characteristic peak is that material is distinctive, it is not easy to coincidence, intensity most strong or stronger peak.
In a kind of implementation method of disclosure one side, the testing sample is the standard items of active component, from
And the feature bar code data storehouse of the standard items can be set up.Various activity can be included in the feature bar code data storehouse
The feature bar code of ingredient standard product, and feature bar code of the various active component standard items under various concentrations.
Wherein, when standard solution of the testing sample for active component, can be by by active component standard items
It is pre-dissolved with chromatographic grade dichloromethane, obtains pre- solution, then the pre- solution to concentration to be measured is diluted with hplc grade methanol and obtain
Obtain the standard solution of active component.
Wherein, when the testing sample is food samples, the pre-treatment step of sample includes：Obtained using liquid-liquid extraction
The first extract of active component, and after through thin-layer chromatography process obtain Active components of food sterling.
In the method that the disclosure is provided, methods described also includes carrying out bioactivity detection, this public affairs to active component
Open and the method for carrying out bioactivity detection had no particular limits, the quantitative determination given activity that can be known in the art into
The method of the bioactivity divided, if the result for accurately reflecting its bioactivity degree is obtained in that, and according to preliminary
Activity Results data set up the potential relation of the hydroxyl and methoxy based structures with bioactivity that represent activity, for example, working as the life
When thing activity is for inoxidizability, it is possible to use DPPH methods, ABTS methods or cell culture processes are detected.
Preferably, the condition of the SERS include laser wavelength be 325nm, 514nm, 532nm,
633nm or 785nm, sweep time is 10-30s, and scanning times are 2-5 times, and wave-number range is 0-4000cm-1。
Preferably, in step sl, the substrate used by SERS is silver-colored dendritic nano particle.
Preferably, the preparation method of silver-colored dendritic nano particle includes：It is 100-200mmol/L's that zinc metal sheet is put into concentration
AgNO3In solution, 20-60s is reacted.
In the specific embodiment of the disclosure, the active component can be Nobiletin, 5- demethyls river dried orange peel
The Polymethoxylated Huangs such as element, 4 '-demethylnobiletin, 5,4 '-dinor- Nobiletin, orange peel element or 5- demethyl orange peel elements
Ketone compounds and polymethoxyflavone glycosides, phenols and terpenes etc.；Specifically, the polymethoxyflavone glycosides can be orange
Skin glycosides, the phenols can be synephrine, and the terpenes are beta carotene.
Second aspect of the disclosure provides a kind of method for detecting Active Ingredient in Food, comprises the following steps：
S1：The feature bar code B of active component in testing sample is obtained using the method described in disclosure one side；
S2：The feature bar code B is compared with the bioactivity of each active component standard items, by testing sample
The chemical constitution of active component be labeled to realize Qualitive test in bar code.
Optionally, methods described also includes detecting the bioactivity of active component standard items, and bioactivity is examined
Survey result to be associated with the feature bar code of the standard items, obtain the chemistry of the active component reflected in feature bar code
Relation between structural information and bioactivity.Specifically, can according to the distribution situation of band in bar code learn activity into
The chemical structure information for dividing, chemical structure information is associated with bioactivity testing result and is compared, so as to learn feature bar
The relation of the distribution situation of band and bioactivity in shape code.
Specifically, the structural information such as hydroxyl and methoxy of compound can be determined according to the position of SERS bar coded strips
Based structures information, so as to judge its bioactivity.
Technical scheme is further described below by embodiment.
Embodiment 1
The present embodiment is detected while being the chemical constitution and antioxidation activity of 6 kinds of polymethoxyflavones
(1) 6 kind of preparation of polymethoxyflavone standard solution
6 kinds of polymethoxyflavones (chemical constitution is as shown in Figure 1) be Nobiletin, 5- demethylnobiletins, 4 '-go first
6 kinds of compounds such as base Nobiletin, 5,4 '-dinor- Nobiletin, orange peel element and 5- demethyl orange peel elements, are from oranges and tangerines
Directly extracted in pericarp separating obtained.This several composition can be pre-dissolved using a small amount of chromatographic grade dichloromethane, then add chromatographic grade
Methanol dilution to concentration is 100ppm solution for standby.
(2) SERS of each polymethoxyflavone functional component is characterized
All of glassware is soaked through being cleaned by ultrasonic ultra-pure water first, it is standby after oven drying.Accurately weigh 3.39g
AgNO3Crystal is dissolved in clean beaker with appropriate ultra-pure water, be transferred to crystallization volumetric flask in be settled to 100ml
It is configured to the AgNO that concentration is 200mmol/L3Solution.The concentrated hydrochloric acid (12mol/L) of 8.5ml is accurately measured using pipette, and
In clean beaker of the injection containing appropriate ultra-pure water, it is stirred continuously and the dilute hydrochloric acid solution that concentration is 1mol/L is configured to after constant volume
It is standby.The zinc metal sheet of formed objects and dilute hydrochloric acid solution are reacted into 1min, is repeated 3 times, and dried after the cleaning of a large amount of ultra-pure waters standby
With.Clean zinc metal sheet is put into AgNO3It is strict to control the reaction time for 40s in solution, it was observed that zinc metal sheet surface covers one layer of black
Or greyish crystals material.By unnecessary AgNO3Solution is slowly outwelled, and the crystalline solid repeatedly rinsed on zinc metal sheet with ultra-pure water
Obtain the dendritic nano active substrates of Ag standby.The 5 μ L dendritic nano particle substrates of silver are accurately pipetted with liquid-transfering gun on clean slide,
40 DEG C of magnetic stirring apparatus heats the moisture for being evaporated top layer, is cooled to room temperature, and SERS detections are carried out to silver-colored dendritic nano particle substrate
Determine its characteristic absorption peak；Nobiletin, 5- demethylnobiletins, 4 '-demethyl river dried orange peel of 2 μ L are pipetted with liquid-transfering gun again
Element, 5,4 '-dinor- Nobiletin, orange peel element, 5- demethyls orange peel 6 kinds of polymethoxyflavone sample solutions of element are accurately added dropwise
In base center position, at room temperature after solvent volatilization completely, the slide is positioned under object lens.Micro- using Horiba swashs
Light Confocal laser-scanning microscopy instrument, the wavelength of laser is 514nm, and sweep time is 10s, scanning times 3 times, and wave-number range is
500-2000cm-1.The second order for carrying out SERS spectrograms by OMNIC and TQ Analyst data processing softwares is changed, in system
In structural relation dimensionality reduction, change at random is eliminated, realize the fixation and recognition of various functional components, every kind of composition is repeated 5 times, obtain phase
Answer SERS second orders conversion spectrogram (as shown in Fig. 2 in figure (1)-(6) represent respectively Nobiletin, 5- demethylnobiletins,
4 '-demethylnobiletin, 5,4 '-dinor- Nobiletin, orange peel element and 5- demethyls orange peel element).
(3) test of each polymethoxyflavone functional component antioxidation activity
The HL-7702 cells RPM I- for containing 10% calf serum, 100U/mL penicillin and 100U/mL streptomysins
1640 culture mediums, in 5%CO2, cellar culture in 37 DEG C of cell culture incubator, 0.25% trypsase, 37 DEG C of digestion 4min, 1：
5 passages, put 37 DEG C, 5%CO2Incubator, continues to cultivate under conditions of relative saturation humidity；The next day change liquid, treat that cell is long extremely
Tested during 90% fusion.Experiment packet and treatment：It is divided into negative control group (the HL-7702 cells without drug-treated),
Experimental group (totally 3 concentration gradients), totally 5 groups, every group of 5 multiple holes.Take the logarithm growth period cell, RPMI-1640 nutrient solutions suspend
And it is 1 × 10 to adjust concentration5Individual/mL, is seeded in 24 orifice plates, 5%CO per the μ L of hole 1002, the culture of 37 DEG C of saturated humidity.Treat that 24 is small
When after cell it is completely adherent, experimental group adds oranges and tangerines functional component to the final concentration of RPMI-1640 complete mediums dilution to distinguish
It it is 2 μm, negative control group adds complete medium, solvent control group adds the training completely of DMSO concentration corresponding with each dosage group
Base is supported, highest DMSO concentration was 0.128% (it is generally acknowledged that DMSO 0.1% does not cause the change of the biological character of cell), often
Hole final volume is 200 μ L.After temperature incubates 4-12h, the H that concentration is 150 μM is added2O2, after continuation temperature incubates 1.5h, supernatant discarded, by examination
Agent cassette method adds the serum free medium containing DCFH-DA and Hoechest33258 fluorescence probes.Cell rinses two after 30min
It is secondary, add serum free medium, fluorescence microplate reader to determine fluorescence intensity, experiment 3 times is repeated, calculate DCFH-DA/
Hoechest33258 fluorescence intensity ratios, the content of reactive oxygen species is characterized with its relative mean fluorescent intensity.It is right with feminine gender
Fluorescence intensity according to hole is 1, calculates H2O2The active oxygen fluorescent value lifting ratio of model group and each sample treatment group.H2O2, river it is old
Pi Su, 5- demethylnobiletin, 4 '-demethylnobiletin, 5,4 '-dinor- Nobiletin, orange peel element and 5- go first
The relative intensity of fluorescence of base orange peel element is respectively 147.5%, 88.3%, 111.0%, 81.9%, 69.3%, 205.4%,
196.7%.(as shown in figure 3,1-6 represents Nobiletin, 5- demethylnobiletins, 4 '-demethyl river dried orange peel respectively in figure
Element, 5,4 '-dinor- Nobiletin, orange peel element and 5- demethyls orange peel element)
(4) conversion of each polymethoxyflavone functional component SERS spectrograms and bar code
The position of the characteristic peak according to each functional component SERS spectrograms and peak intensity, to each characteristic peak wave trough position ± 5cm-1
The peak intensity in region carry out the cumulative peak intensity that obtains of peak intensity value and add and be worth, force and be worth maximum characteristic peak as base using middle peak of spectrogram
Quasi- peak is normalized, and the bandwidth of the corresponding bar code after being normalized is set to 10 pounds, according to formula：Bar code
Strip width value=a × peak intensity adds and is worth to corresponding a values, the peak intensity of remaining each characteristic peak add and value and a values product i.e.
It is the strip width value of corresponding bar code.By taking Nobiletin as an example, in its SERS spectrogram, most strong peak appears in 1610cm-1,
To this peak ± 5cm-1Region, i.e. 1605-1615cm-1In the range of peak intensity sum up, its add and be worth about 30000, to this
Value is normalized, and the width of bar coded strip is set into 10 pounds, and it is 1/3000 to obtain a values.Nobiletin is other
Characteristic peak peak intensity adds and is worth the strip width value for being multiplied by a as its corresponding bar code.The bar code strip bandwidth of other compounds
Value is calculated in the method.Drawn according to the distributing position and bar coded strip width value that second order changes each characteristic peak in spectrogram
The feature bar code of active component.
(5) conversion of each functional component antioxidation activity and bar code
The position of the hydroxyl in the antioxidation activity of each functional component and its chemical constitution and quantity are closely related, with each work
The fluorescence intensity ratio of property composition as its anti-oxidant observation index, according to antioxidation activity data result determine all kinds of effects into
The antioxidation activity for dividing is strong and weak, while the feature bar code opening relationships with reflection chemical constitution, in 1328-1357cm-1And
1417-1428cm-1In the range of have hydroxyl vibration peak PMFs show antioxidation activity higher, can see in bar code
Corresponding feature marks band (as shown in figure 4, black stripe represents high anti-oxidation active structure characteristic bands, grey bar in Fig. 4
Band represents the characteristic bands for structural characterization；In figure (1)-(6) represent respectively Nobiletin, 5- demethylnobiletins, 4 '-
Demethylnobiletin, 5,4 '-dinor- Nobiletin, orange peel element and 5- demethyls orange peel element).
Embodiment 2
Polymethoxyflavone glycosides, phenols and terpenes based food functional component chemical constitution (chemical constitution is as shown in Figure 1) with
Detected while bioactivity.
(1) preparation of polymethoxyflavone glycosides, phenols and terpenes based food functional component standard solution
Aurantiamarin is a kind of Flavonoid substances with glucosides, and dissolubility is similar with polymethoxyflavone, using chromatogram two
Chloromethanes is pre-dissolved, then standby to 100ppm with chromatogram methanol dilution.Aldehydes matter synephrine is a kind of water miscible oranges and tangerines work(
Effect composition, the testing sample solution that 100ppm is configured to by the way of ultra-pure water and heating stirring is standby.Beta carotene is one
Isoprene based food functional component is planted, is mainly dissolved in the organic solvents such as chloroform, this experiment is mainly pre- using dichloromethane
It is 100ppm standby to be diluted to concentration with n-hexane again after dissolving.
(2) SERS of polymethoxyflavone glycosides, phenols and terpenes based food functional component is characterized
Specific experiment flow it is identical with embodiment 1 (second order changes collection of illustrative plates as shown in Fig. 2 wherein, and (7) refer to aurantiamarin,
(8) synephrine is referred to, (9) refer to beta carotene).
(3) polymethoxyflavone glycosides, phenols and terpenes based food functional component biological activity determination
Specific experiment flow is same as Example 1, the relative mean fluorescent intensity of orange peel mandarin orange, synephrine and beta carotene
Respectively 93.1%, 127.9% and 74.0%, control group H2O2For 147.5% (as shown in figure 3, wherein, 7 refer to aurantiamarin, 8
Synephrine is referred to, 9 refer to beta carotene).
(4) conversion of polymethoxyflavone glycosides, phenols and terpenes based food functional component bar code
The position of the characteristic peak of the SERS spectrograms according to each functional component and peak intensity, to each characteristic peak wave trough position ±
5cm-1The peak intensity in region carry out the cumulative peak intensity that obtains of peak intensity value and add and be worth, force and be worth the characteristic peak work of maximum with middle peak of spectrogram
On the basis of peak be normalized, and the bandwidth of the corresponding bar code after being normalized is set to 10 pounds, the peak of characteristic peak
Force and value is the bandwidth numerical value of corresponding bar code with the product of a values.Specific implementation process and the method described in embodiment 1
It is identical.
Determine that the antioxidation activity of each functional component is strong and weak according to antioxidation activity data result, at the same with reactive chemistry knot
The bar code of structure is corresponded to set up the relation of bar code and antioxidation activity, specifically：As polymethoxyflavone glycosides generation
The aurantiamarin of table is because in 1320-1327cm-1In the range of there is the representative band of hydroxyl in representative structure and be presented higher anti-
Oxidation activity, as the synephrine of phenols representative in 1324cm-1And 1407cm-1Place has the band of representation hydroxy and height is presented
Activity, and the beta carotene represented as terpenes because in structure multiple isoprene structures and in 1677cm-1Place occurs special
Different in nature band and antioxidation activity higher is presented, can see in bar code corresponding feature mark band (Fig. 4, wherein, it is black
Vitta band represents sign high anti-oxidation living features band, and gray bars represent the characteristic bands for structural characterization；(7) refer to
Aurantiamarin, (8) refer to synephrine, and (9) refer to beta carotene).
Describe the preferred embodiment of the disclosure in detail above in association with accompanying drawing, but, the disclosure is not limited to above-mentioned reality
The detail in mode is applied, in the range of the technology design of the disclosure, various letters can be carried out with technical scheme of this disclosure
Monotropic type, these simple variants belong to the protection domain of the disclosure.
It is further to note that each particular technique feature described in above-mentioned specific embodiment, in not lance
In the case of shield, can be combined by any suitable means, in order to avoid unnecessary repetition, the disclosure to it is various can
The combination of energy is no longer separately illustrated.
Additionally, can also be combined between a variety of implementation methods of the disclosure, as long as it is without prejudice to originally
Disclosed thought, it should equally be considered as disclosure disclosure of that.
Claims (10)
1. a kind of method for characterizing Active Ingredient in Food, comprises the following steps：
SERS is carried out to testing sample, second order conversion is carried out to Raman scattering collection of illustrative plates and peak-fit processing is obtained two
Rank is changed spectrogram and determines the characteristic peak of active component, to the characteristic peak wave trough position ± 5cm of the active component-1Region enter
The cumulative peak intensity that obtains of row peak intensity value adds and is worth, and peak intensity is added as reference peaks and value is carried out using peak intensity plus with the characteristic peak of value maximum
Normalized, special value X is set to by the bar coded strip width value of reference peaks, and the value of a is calculated using formula I, profit
The bar coded strip width value of all characteristic peaks is calculated with a values, the distribution position of each characteristic peak in spectrogram is changed according to second order
Put and the bar coded strip width value draws the feature bar code of the active component, in the feature bar code, each
Bar coded strip all characterizes a chemical structure information of the active component；
Formula I：Bar coded strip width value=a × peak intensity adds and is worth.
2. method according to claim 1, it is characterised in that methods described also includes, will characterize the active component
The bar coded strip of particular organisms activity related chemical structure is defined as the activity band of the bioactivity, by activity band face
Color 1 is marked, by other colors for having with color 1 cognizable difference of other bar coded strips in addition to the activity band
Mark.
3. method according to claim 2, it is characterised in that the span of a is 1/10000-1/1000, described specific
Numerical value X is 10 pounds.
4. the method according to any one in claim 1-3, it is characterised in that the testing sample is active component
Standard items, set up the feature bar code data storehouse of the standard items.
5. method according to claim 4, it is characterised in that the condition of the SERS includes laser
Wavelength is 325nm, 514nm, 532nm, 633nm or 785nm, and sweep time is 10-30s, and scanning times are 2-5 times, wave number model
It is 0-4000cm to enclose-1。
6. the method according to any one in claim 1-3 and 5, it is characterised in that used by SERS
Substrate be silver-colored dendritic nano particle.
7. method according to claim 6, it is characterised in that the preparation method of silver-colored dendritic nano particle includes：By zinc metal sheet
It is put into the AgNO that concentration is 100-200mmol/L320-60s is reacted in solution.
8. the method according to any one in claim 1-3,5 and 7, it is characterised in that the active component includes class
Isoprene derivatives, aldehydes matter, the derivative based on protein or amino acid, carbohydrate and its derivative and
Polyunsaturated fatty acid.
9. a kind of method for detecting Active Ingredient in Food, comprises the following steps：
S1：The feature bar code of active component in testing sample is obtained using the method described in any one in claim 1-8
B；
S2：The feature bar code B is compared with the bioactivity of each active component standard items, by the work in testing sample
The chemical constitution of property composition is labeled to realize Qualitive test in bar code.
10. method according to claim 9, it is characterised in that methods described also includes the life to active component standard items
Thing activity is detected, bioactivity testing result and Structure-activity analysis result is entered with the feature bar code of the standard items
Row is compared, and obtains the relation between the chemical structure information and bioactivity of the active component reflected in feature bar code.
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| CN110068565A (en) * | 2019-06-06 | 2019-07-30 | 长江师范学院 | The application of SERS sensing chip and its detection method and preparation method |
| WO2021053409A1 (en) * | 2019-09-20 | 2021-03-25 | King Abdullah University Of Science And Technology | Encoding raman spectral data in optical identification tags for analyte identification |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110068565A (en) * | 2019-06-06 | 2019-07-30 | 长江师范学院 | The application of SERS sensing chip and its detection method and preparation method |
| WO2021053409A1 (en) * | 2019-09-20 | 2021-03-25 | King Abdullah University Of Science And Technology | Encoding raman spectral data in optical identification tags for analyte identification |
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