CN106290304A - A kind of method for quick of plant polyphenol - Google Patents
A kind of method for quick of plant polyphenol Download PDFInfo
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- CN106290304A CN106290304A CN201610810319.XA CN201610810319A CN106290304A CN 106290304 A CN106290304 A CN 106290304A CN 201610810319 A CN201610810319 A CN 201610810319A CN 106290304 A CN106290304 A CN 106290304A
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- G—PHYSICS
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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
- G01N21/65—Raman scattering
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- G—PHYSICS
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Abstract
The present invention relates to the method for quick of a kind of plant polyphenol.Its technical scheme is: 1) add sodium citrate in tannic acid solution, stirs evenly post-heating and adds silver ammino solution under conditions of stirring, obtaining the elargol of modification；2) with the plant polyphenol standard solution of the differently configured gradient concentration of distilled water, plant polyphenol standard solution is mixed with elargol, centrifugal collecting precipitation part, and utilize laser Raman spectrometer to test, linear equation between plant polyphenol Raman spectrum absorption intensity and plant polyphenol concentration, i.e. y=kx+b；3) use step 2) described in method, utilize laser Raman spectrometer that the plant polyphenol solution of unknown concentration is tested, obtain plant polyphenol Raman spectrum absorption intensity under corresponding wave number, extrapolate the concentration of plant polyphenol according to linear equation.The method of the present invention has that pretreatment sample size simple, required is few, the test analysis time is short, sensitivity advantages of higher, and detection limit reaches 10ng/L.
Description
Technical field
The present invention relates to the method for quick of a kind of plant polyphenol.Specifically with nanometer silver as substrate, by drawing
The method for quick of a kind of plant polyphenol that graceful spectrographic technique is carried out.
Background technology
Plant polyphenol (Plant polyphenol) has another name called vegatable tannin (Vegetable tannin), in plant
Complicated phenols secondary metabolites, has polyhydric phenols structure, is primarily present in the skin of plant, root, leaf, fruit, containing in plant
Amount is only second to cellulose, hemicellulose and lignin.
Although polyphenol is widely present in plant food, but its content and structure of in different foods has the biggest difference
Different.And, in plant, the content of polyphenol is affected by factors, such as maturity, kind, the course of processing and condition of storage etc..
The method using more detection plant polyphenol kind at present in the world is chromatography, especially high-resolution gas chromatogram
In conjunction with high resolution mass spectrum multiple techniques.The method needs complicated sample pretreatment process, and test period is longer, and requires have
Accurate instrument, good experimental situation, through the operator of professional training and qualitative, quantitative standard sample etc., use
This method detection plant polyphenol kind is costly.
Surface enhanced raman spectroscopy technology (SERS) is few, to sample without destructiveness, Er Qieling to the requirement of detection sample
The advantage that sensitivity is high.Being simultaneously based on the best development of laser technology, computer technology, particularly nanotechnology is continuous
Widen with increasingly mature, constantly provide variation, active substrate that enhancement effect is good for SERS technology, make the application of SERS technology
Field the most constantly extends.Due to the non-destructive of sample and the peak position resolution energy to the not dependent of exciting light and finger-print type
Power, thus the trace detection of material can be realized.
Utilization shows to strengthen Raman spectroscopy detection by quantitative plant polyphenol, there is not yet relevant report both at home and abroad, this field
It is still blank at present.
Summary of the invention
In order to overcome that the detection of existing plant polyphenol exists costly, the problem such as complicated, the time-consuming length of pretreatment, the present invention's
Purpose is to provide the method for quick of a kind of plant polyphenol, has easily operated, highly sensitive, can save a large amount of economy and time
Between the advantage of cost.
To achieve these goals, present invention employs following technical scheme:
1) in tannic acid solution, sodium citrate is added, after stirring, heating and addition and tannic acid under conditions of stirring
The silver ammino solution of solution same volume, reaction obtains tannic acid modified elargol;
2) with the plant polyphenol standard solution of the differently configured gradient concentration of distilled water, respectively the plant polyphenol of different gradient concentrations
Standard solution is by the volume ratio of 1:1 and step 1) the tannic acid modified elargol that obtains uniformly mixes, centrifugation, collects precipitation
Part, and utilize laser Raman spectrometer to use no absorbing light spectral peak to test, obtain the SERS spectra of plant polyphenol
Figure, obtain plant polyphenol Raman spectrum absorption intensity (y) and plant polyphenol concentration (x) between relation, set up Raman spectrum inhale
Receive the linear equation between intensity and plant polyphenol concentration, i.e. y=kx+b;
3) use step 2) described in method, utilize laser Raman spectrometer to use step 2) corresponding absorbing light spectral peak is not to
Know that the plant polyphenol solution of concentration is tested, obtain plant polyphenol Raman spectrum absorption intensity under corresponding wave number, according to step
2) plant polyphenol is extrapolated in the linear equation between the plant polyphenol Raman spectrum absorption intensity and the plant polyphenol concentration that obtain
Concentration；Choose in 5 data immediate 3 to average, be the concentration of the plant polyphenol of unknown concentration.
Above-mentioned steps 1) in: the mass fraction of described tannic acid solution is 0.05%-0.5%；Described sodium citrate
Consumption is the 30-50% of tannic acid quality；Described heating-up temperature is preferably 60-80 DEG C；The concentration of described silver ammino solution is
5mmol/L-20mmol/L；Described reaction, its time is 2-3 hour.
Above-mentioned steps 2) in: the plant polyphenol standard solution of described different gradient concentrations is the concentration using distilled water configuration
Scope is the plant polyphenol standard solution of 6 gradient concentrations of Concentraton gradient at equal intervals between 5ng/L-50ng/L.Described suction
Receiving spectrum peak is wave number 1420 cm-1、1409 cm-1、1254 cm-1、1110 cm-1With 1102 cm-1。
Above-mentioned steps 3) in: before the plant polyphenol solution of described unknown concentration is tested, with distilled water to the unknown
The plant polyphenol solution of concentration carries out constant volume.
Advantages of the present invention is: 1) method of the present invention have pretreatment sample size simple, required few (as long as 0.5ml),
Test analysis time short (less than 30 minutes), sensitivity advantages of higher, detection limit reaches 10ng/L.
2) use the colloidal sol prepared of tannic acid to have preparation is simple, material therefor environmental protection, colloidal sol long shelf-life (room temperature
Lower preservation was up to 8 months) etc. advantage.
3) utilize the multiformity of the molecular structure of tannic acid, the different-shape SERS with size can be prepared with receiving
Rice SERS substrate；Utilize the hydroxyl reproducibility of tannic acid, the effect preparing SERS nanometer silver seed can be played, reduce reducing agent
Consumption, reaches the effect preferably controlling SERS substrate pattern with particle diameter；Utilize the polymer characteristics of tannic acid, be possible to prevent nanometer
The reunion of silver, it is achieved that the advantages such as the long-time preservation (storage life reaches more than 8 months) at the bottom of SERS nano silver-group.Solve existing
Prepare with the presence of technology that the pattern of SERS nanometer silver is wayward, poor reproducibility, product are single, storage life is short (about 3 months)
Etc. shortcoming, it can be seen that storage life at the bottom of the SERS nano silver-group of the present invention is more than two sesquialters of prior art.
4) having higher sensitivity at the bottom of nano silver-group prepared by the present invention, the sensitivity of the present invention is at least 10ng/L,
It is 100 times of prior art, as long as 1st/50th of testing cost liquid phase-mass spectrometry instrument assay method.
Accompanying drawing explanation
Fig. 1 is the transmission electron microscope picture of elargol tannic acid modified in embodiment 1.
Fig. 2 is the surface enhanced raman spectroscopy figure of 6 the gradient concentration plant polyphenols recorded in embodiment 1.
Fig. 3 is to record 6 gradient concentration plant polyphenol surface enhanced raman spectroscopy in embodiment 1 at 1420 cm-1At wave number
The standard working curve figure of relation between absorption peak strength and concentration.
Fig. 4 is to record 6 gradient concentration plant polyphenol surface enhanced raman spectroscopy in embodiment 1 at 1409 cm-1At wave number
The standard working curve figure of relation between absorption peak strength and concentration.
Fig. 5 is to record 6 gradient concentration plant polyphenol surface enhanced raman spectroscopy in embodiment 1 at 1254 cm-1At wave number
The standard working curve figure of relation between absorption peak strength and concentration.
Fig. 6 is to record 6 gradient concentration plant polyphenol surface enhanced raman spectroscopy in embodiment 1 at 1110 cm-1At wave number
The standard working curve figure of relation between absorption peak strength and concentration.
Fig. 7 is to record 6 gradient concentration plant polyphenol surface enhanced raman spectroscopy in embodiment 1 at 1102 cm-1At wave number
The standard working curve figure of relation between absorption peak strength and concentration.
Detailed description of the invention
Below with reference to embodiment, the present invention will be further described with accompanying drawing.
Embodiment 1
(1) tannic acid of configuration quality mark 0.2%, adds the sodium citrate that quality is tannic acid 37.5%, after stirring, rises
High to 75 DEG C of temperature, adding under conditions of stirring with tannic acid solution same volume concentration is 10mmol/L silver ammino solution, 75
React 3 hours at a temperature of DEG C, obtain tannic acid modified elargol.
(2) 6 gradient concentrations (10ng/L, 15ng/L, 20ng/L, 25ng/L, 30ng/L, 35ng/L) are configured with distilled water
Plant polyphenol standard solution, the plant polyphenol standard solution of 1ml variable concentrations is uniformly mixed with the elargol of 1ml, centrifugal point
From, collect sediment fraction, and utilize laser Raman spectrometer to test, obtain the SERS spectra figure of plant polyphenol, see Fig. 1.
Obtaining absorption spectrum spike number is 1420 cm-1、1409 cm-1、1254 cm-1、1110 cm-1With 1102 cm-1's
Relation between plant polyphenol Raman spectrum absorption intensity (y) and plant polyphenol concentration (x), sets up Raman spectrum absorption intensity
Y the linear equation between () and plant polyphenol concentration (x) is as follows:
1420 cm-1Linear equation is y=699.71x-1304 linearly dependent coefficient R2=0.9918(is shown in Fig. 3)
1409 cm-1Linear equation is y=8122.5x-32380 linearly dependent coefficient R2=0.9888(is shown in Fig. 4)
1254 cm-1Linear equation is y=533.65x 904.19 linearly dependent coefficient R2=0.9919(is shown in Fig. 5)
1110 cm-1Linear equation is y=470.39x 781.64 linearly dependent coefficient R2=0.9904(is shown in Fig. 6)
1102 cm-1Linear equation is y=551x 1248.3 linearly dependent coefficient R2=0.9927(is shown in Fig. 7)
(3) with distilled water, the plant polyphenol solution of unknown concentration is carried out constant volume, the plant polyphenol solution of unknown concentration with etc.
The step 1 of volume) the tannic acid modified elargol that obtains uniformly mixes, centrifugation, and collect sediment fraction, utilize LR laser raman
The plant polyphenol solution of unknown concentration is tested by spectrogrph, and obtaining wave number is 1420 cm-1、1409 cm-1、1254 cm-1、
1110 cm-1With 1102 cm-1Plant polyphenol Raman spectrum absorption intensity be respectively 12270. 4,126821,9501.9,
9237.7、9964.55.According to the formula in step (2), the concentration being calculated plant polyphenol is 19.4 ng/L, 19.6 ng/
L, 19.5 ng/L, 21.3 ng/L, 20.35 ng/L, average, and the concentration obtaining plant polyphenol to be measured is 20.0ng/L.
Embodiment 2
(1) tannic acid of configuration quality mark 0.5%, adds the sodium citrate that quality is tannic acid 45%, after stirring, raises
To 70 DEG C of temperature, adding under conditions of stirring with tannic acid solution same volume concentration is 17.5mmol/L silver ammino solution, 75
React 2.5 hours at a temperature of DEG C, obtain tannic acid modified elargol.
(2) 6 gradient concentrations (10ng/L, 15ng/L, 20ng/L, 25ng/L, 30ng/L, 35ng/L) are configured with distilled water
Plant polyphenol standard solution, the plant polyphenol standard solution of 1ml variable concentrations is uniformly mixed with the elargol of 1ml, centrifugal point
From, collect sediment fraction, and utilize laser Raman spectrometer to test, obtain the SERS spectra figure of plant polyphenol.
Obtaining absorption spectrum spike number is 1420 cm-1、1409 cm-1、1254 cm-1、1110 cm-1With 1102 cm-1's
Relation between plant polyphenol Raman spectrum absorption intensity (y) and plant polyphenol concentration (x), sets up Raman spectrum absorption intensity
Y the linear equation between () and plant polyphenol concentration (x) is as follows:
1420 cm-1Linear equation is y=722.28x-1587 linearly dependent coefficient R2 = 0.9968
1409 cm-1Linear equation is y=8358.7x-38957 linearly dependent coefficient R2 = 0.9987
1254 cm-1Linear equation is y=587.39x 874.32 linearly dependent coefficient R2 = 0.9934
1110 cm-1Linear equation is y=467.2x 989.31 linearly dependent coefficient R2 = 0.9945
1102 cm-1Linear equation is y=565x 1148.3 linearly dependent coefficient R2 = 0.9972
(3) with distilled water, the plant polyphenol solution of unknown concentration is carried out constant volume, the plant polyphenol solution of unknown concentration with etc.
The step 1 of volume) the tannic acid modified elargol that obtains uniformly mixes, centrifugation, and collect sediment fraction, utilize LR laser raman
The plant polyphenol solution of unknown concentration is tested by spectrogrph, and obtaining wave number is 1420 cm-1、1409 cm-1、1254 cm-1、
1110 cm-1With 1102 cm-1Plant polyphenol Raman spectrum absorption intensity be respectively 16903.4,183384.4,13754.2,
11765.3、13372.2.According to the linear equation formula in step (2), the concentration being calculated plant polyphenol is: 25.6ng/
L, 26.6ng/L, 25.1ng/L, 27.3ng/L, 25.7ng/L, average, and the concentration obtaining plant polyphenol to be measured is
26.1ng/L。
Embodiment 3
(1) tannic acid of configuration quality mark 0.1%, adds the sodium citrate that quality is tannic acid 40%, after stirring, raises
To 78 DEG C of temperature, adding under conditions of stirring with tannic acid solution same volume concentration is 7.5mmol/L silver ammino solution, 72 DEG C
At a temperature of react 2.5 hours, obtain tannic acid modified elargol.
(2) 6 gradient concentrations (10ng/L, 15ng/L, 20ng/L, 25ng/L, 30ng/L, 35ng/L) are configured with distilled water
Plant polyphenol standard solution, the plant polyphenol standard solution of 1ml variable concentrations is uniformly mixed with the elargol of 1ml, centrifugal point
From, collect sediment fraction, and utilize laser Raman spectrometer to test, obtain the SERS spectra figure of plant polyphenol.
Obtaining wave number is 1420 cm-1、1409 cm-1、1254 cm-1、1110 cm-1With 1102 cm-1Plant polyphenol draw
Relation between graceful spectral absorption intensity and plant polyphenol concentration, sets up Raman spectrum absorption intensity (y) and plant polyphenol concentration
X the linear equation between () is as follows:
1420 cm-1Linear equation is y=783.25x-1289 linearly dependent coefficient R2 = 0.9938
1409 cm-1Linear equation is y=8324.6x-34785 linearly dependent coefficient R2 = 0.9878
1254 cm-1Linear equation is y=533.65x 904.19 linearly dependent coefficient2 = 0.9932
1110 cm-1Linear equation is y=470.39x 781.64 linearly dependent coefficient R2 = 0.9948
1102 cm-1Linear equation is y=539x 1357.8 linearly dependent coefficient R2 = 0.9967
(3) with distilled water, the plant polyphenol solution of unknown concentration is carried out constant volume, the plant polyphenol solution of unknown concentration with etc.
The step 1 of volume) the tannic acid modified elargol that obtains uniformly mixes, centrifugation, and collect sediment fraction, utilize LR laser raman
The plant polyphenol solution of unknown concentration is tested by spectrogrph, and obtaining wave number is 1420 cm-1、1409 cm-1、1254 cm-1、
1110 cm-1With 1102 cm-1Plant polyphenol Raman spectrum absorption intensity be respectively 24009.9,237429.4,16439.4,
14317.9、16321.4.According to the linear equation formula in step (2), the concentration being calculated plant polyphenol is: 32.3ng/
L, 32.7ng/L, 32.5ng/L, 32.1ng/L, 32.8ng/L, average, and the concentration obtaining plant polyphenol to be measured is
32.5ng/L。
The foregoing is only embodiments of the invention, not thereby limit the scope of the claims of the present invention, every utilize this
Equivalent structure or equivalence flow process that bright description is made convert, or are directly or indirectly used in other relevant technology necks
Territory, is the most in like manner included in the scope of patent protection of the present invention.
Claims (8)
1. a method for quick for plant polyphenol, is characterized in that:
1) in tannic acid solution, sodium citrate is added, after stirring, heating and addition and tannic acid under conditions of stirring
The silver ammino solution of solution same volume, reaction obtains tannic acid modified elargol；
2) with the plant polyphenol standard solution of the differently configured gradient concentration of distilled water, respectively the plant polyphenol of different gradient concentrations
Standard solution is by the volume ratio of 1:1 and step 1) the tannic acid modified elargol that obtains uniformly mixes, centrifugation, collects precipitation
Part, and utilize laser Raman spectrometer to use no absorbing light spectral peak to test, obtain the SERS spectra of plant polyphenol
Figure, obtains the relation between plant polyphenol Raman spectrum absorption intensity and plant polyphenol concentration, sets up Raman spectrum absorption intensity
And the linear equation between plant polyphenol concentration, i.e. y=kx+b；
3) use step 2) described in method, utilize laser Raman spectrometer to use step 2) corresponding absorbing light spectral peak is not to
Know that the plant polyphenol solution of concentration is tested, obtain plant polyphenol Raman spectrum absorption intensity under corresponding wave number, according to step
2) plant polyphenol is extrapolated in the linear equation between the plant polyphenol Raman spectrum absorption intensity and the plant polyphenol concentration that obtain
Concentration；Choose in 5 data immediate 3 to average, be the concentration of the plant polyphenol of unknown concentration.
The method for quick of a kind of plant polyphenol the most according to claim 1, is characterized in that the list described in step 1)
The mass fraction of thujic acid solution is 0.05%-0.5%.
The method for quick of a kind of plant polyphenol the most according to claim 1, is characterized in that adding described in step 1)
Hot temperature is preferably 60-80 DEG C.
The method for quick of a kind of plant polyphenol the most according to claim 1, is characterized in that silver ammonia described in step 1)
The concentration of solution is 5mmol/L-20mmol/L.
The method for quick of a kind of plant polyphenol the most according to claim 1, it is characterized in that described in step 1) is anti-
Should, its time is 2-3 hour.
The method for quick of a kind of plant polyphenol the most according to claim 1, is characterized in that step 2) described in different
The plant polyphenol standard solution of gradient concentration be use distilled water configuration concentration range between 5ng/L-50ng/L at equal intervals
The plant polyphenol standard solution of 6 gradient concentrations of Concentraton gradient.
The method for quick of a kind of plant polyphenol the most according to claim 1, is characterized in that step 2) described in suction
Receiving spectrum peak is wave number 1420 cm-1、1409 cm-1、1254 cm-1、1110 cm-1With 1102 cm-1。
The method for quick of a kind of plant polyphenol the most according to claim 1, is characterized in that above-mentioned steps 3) described in
The plant polyphenol solution of unknown concentration test before, it is fixed to carry out the plant polyphenol solution of unknown concentration with distilled water
Hold.
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| CN106841637A (en) * | 2017-02-21 | 2017-06-13 | 南昌大学 | A kind of Nano silver grain delustring immuno-chromatographic test paper strip for detecting small-molecule substance |
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