CN109324111A - A method of the identification eupatorium true and false - Google Patents
A method of the identification eupatorium true and false Download PDFInfo
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- CN109324111A CN109324111A CN201811311930.3A CN201811311930A CN109324111A CN 109324111 A CN109324111 A CN 109324111A CN 201811311930 A CN201811311930 A CN 201811311930A CN 109324111 A CN109324111 A CN 109324111A
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Abstract
The invention discloses a kind of methods for identifying the eupatorium true and false.This method extracts herb sample with different solvents and obtains different extract liquors, it is modified respectively in electrode surface, differential pulse voltammetry scanning is carried out to modified electrode after to be dried, electrochemistry finger-print is converted by obtained electrochemical signals, coincide by comparing sample to be tested and the whether high similarity of standard eupatorium electrochemistry finger-print and judges whether sample to be tested is eupatorium certified products.The advantages that present invention has identification method simple and easy, at low cost, and accuracy is high.
Description
Technical field
The invention belongs to Analysis of Chinese Traditional Medicine technical fields, more particularly to a kind of method for identifying the eupatorium true and false.
Background technique
Chinese Pharmacopoeia (version in 2015) regulation eupatorium certified products is composite family eupatorium (Eupatorium fortune Turcz.)
Dry aerial parts.Eupatorium and Herba Lycopi are conventional Chinese medicine, and the two effect is different.Eupatorium mildly bitter flavor is cold in nature, have antipyretic clear heat,
Dispelling damp-cold from the stomach and improving appetite and digestion, the effect of preventing or arresting vomiting.Herba Lycopi (Lycopus lucidus Turcz.vat.hirtus Regel) bitter slightly warm in nature,
There are promoting blood circulation, the effect of inducing diuresis for removing edema.
Since ancient times, just often there are aliasing in various regions to eupatorium and Herba Lycopi, first is that due in Chinese herbal medicine homonym show
As very common, second is that being difficult to after herbal medicine is broken with identifying by naked eye.Nowadays, adulterant is more in eupatorium commodity, in addition to Herba Lycopi, often
Also Eupatorium adenophorum, Herba Serissae and the Eupatorium lindleyanum etc. seen.To ensure medical effect and drug safety, the express delivery of eupatorium is at a low price
Identification has very important application space.It in the prior art can be by ITS2 bar code sequence method to eupatorium and its mixed adulterant
It is differentiated, but amplification and sequencing of this method based on DNA sequence dna, it is clear that trivial operations and higher cost.Therefore, develop
A kind of technology of quick identification eupatorium is highly desirable.
To sum up, the present invention in view of the above-mentioned defects in the prior art and the market demand, develop it is a kind of from Chinese medicine eupatorium and
The method that eupatorium certified products is identified in its mixed adulterant, this method utilize Electrochemistry Knowledge, are realized by easy-to-use method
The quick identification of eupatorium certified products.
Summary of the invention
The purpose of the present invention is to provide a kind of methods for identifying the eupatorium true and false.The present invention has identification method simply easy
Row, at low cost, the advantages that accuracy is high.
To achieve the above object, the present invention takes following technical scheme:
A method of the identification eupatorium true and false includes the following steps:
(1) fragment of tissue of sample to be tested is added separately to extract in n kind different solvents, obtains n kind extract liquor；
Wherein n >=2.Sample to be tested can be eupatorium and its fresh herbal or drying herbal medicine of mixed adulterant.
(2) n kind extract liquor is coated on screen printing electrode surface with liquid-transfering gun respectively, obtains n kind after its drying and repairs
Adorn electrode.
(3) the n kind modified electrode is respectively placed in progress voltammetry scanning in phosphate buffer, by n kind modified electrode
The multi-loop electric chemical signal respectively obtained constructs to obtain the two-dimentional electrochemistry finger-print of sample to be tested.
(4) the standard two-dimensional electrochemistry finger-print of the electrochemistry finger-print of sample to be tested and eupatorium certified products is carried out pair
Than if the electrochemistry finger-print of sample to be tested is similar to standard diagram height, sample to be tested is eupatorium；Conversely, being then adulterant.
Preferably, the quality of the fragment of tissue in the step (1) and the volume ratio of each solvent are (0.1~1.5) g:
(0.5~4) ml.
Preferably, the different solvents in the step (1) include dimethylformamide or methanol.
Preferably, coating sample size used is 1~5 μ l in the step (2).
Preferably, the drying mode in the step (2) is 60 DEG C of heating, dryings.
Preferably, the voltammetric scan mode in the step (3) is differential pulse voltammetry scanning.
Preferably, the phosphate buffer in the step (3) is the mixed solution of potassium dihydrogen phosphate and sodium dihydrogen phosphate, respectively
From concentration be respectively 0.05~0.2mol/L.
Preferably, the sweep interval of pulse voltammetric scan is 1.2V~-0.2V in the step (3).
Preferably, in the step (3) two-dimentional electrochemistry finger-print by any one group of differential pulse voltammetry scanning figure
The current value of spectrum is as fingerprint image X-axis data, and the current value of another component pulse voltammetry scanning spectra is as fingerprint image Y-axis number
According to gained.
The present invention provides one kind based on electrochemical analysis, is to implement the eupatorium certified products of object with eupatorium and its adulterant
Discrimination method.Different extract liquors are obtained with different solvents extraction herb sample, it is modified respectively in electrode surface, after to be dried
Differential pulse voltammetry scanning is carried out to modified electrode, electrochemistry finger-print is converted by obtained electrochemical signals, passes through ratio
It coincide compared with sample to be tested and the whether high similarity of standard eupatorium electrochemistry finger-print and judges whether sample to be tested is eupatorium certified products.
Differential pulse voltammetry scanning is to be summed it up with the fixed pulse of linear potential or ladder potential and amplitude as pumping signal.Will
Using before pulse and pulse latter stage, to electric current two sub-samplings of progress.This kind of scanning mode possesses than normal pulsed voltammetric scan
Lower background current value, detection sensitivity is higher, and detection limit is lower, is more suitable for the electrolyte environment of low concentration, favorably
The signal measuring of trace materials in plant.
The present invention has following technical characterstic:
1) identification method provided by the invention is simple and easy, is not necessarily to expensive reagent and instrument, reduces appraisal cost.
2) discrimination method of the invention can fresh goods to eupatorium and its mixed adulterant and dry product identify.
Detailed description of the invention
Fig. 1 is the standard two-dimensional electrochemistry finger-print that differential pulse voltammetry scans resulting fresh eupatorium.
Fig. 2 is the two-dimentional electrochemistry finger-print that differential pulse voltammetry scans resulting fresh eupatorium to be measured.
Fig. 3 is the two-dimentional electrochemistry finger-print that differential pulse voltammetry scans resulting fresh Herba Lycopi.
Fig. 4 is the two-dimentional electrochemistry finger-print that differential pulse voltammetry scans resulting fresh Eupatorium adenophorum.
Fig. 5 is the two-dimentional electrochemistry finger-print that differential pulse voltammetry scans resulting fresh Herba Serissae.
Fig. 6 is the two-dimentional electrochemistry finger-print that differential pulse voltammetry scans resulting fresh Eupatorium lindleyanum.
Fig. 7 is the standard two-dimensional electrochemistry finger-print that differential pulse voltammetry scans resulting drying eupatorium.
Fig. 8 is the two-dimentional electrochemistry finger-print that differential pulse voltammetry scans resulting drying eupatorium to be measured.
Fig. 9 is the two-dimentional electrochemistry finger-print that differential pulse voltammetry scans resulting drying Herba Lycopi.
Figure 10 is the two-dimentional electrochemistry finger-print that differential pulse voltammetry scans resulting drying Eupatorium adenophorum.
Figure 11 is the two-dimentional electrochemistry finger-print that differential pulse voltammetry scans resulting drying Herba Serissae.
Figure 12 is the two-dimentional electrochemistry finger-print that differential pulse voltammetry scans resulting drying Eupatorium lindleyanum.
Figure 13 is the standard two-dimensional electrochemistry finger-print that normal pulsed voltammetry scans resulting fresh eupatorium.
Figure 14 is the two-dimentional electrochemistry finger-print that normal pulsed voltammetry scans resulting fresh eupatorium to be measured.
Specific embodiment
Following specific embodiments are the further explanations to method provided by the invention and technical solution, but are not construed as
Limitation of the present invention.
Embodiment 1: the detection to fresh herbal
(1) fresh Chinese medicine eupatorium the building of eupatorium certified products standard electrochemical finger-print: is shredded into tissue with scissors
Fragment, respectively throw 0.5g fragment of tissue into 2mL dimethylformamide and methanol solution, sufficiently oscillation after, obtain two parts it is fresh
The eupatorium extract liquor in dimethylformamide and methanol respectively.Above-mentioned 2 μ L of extract liquor is respectively taken, is respectively coated with liquid-transfering gun in silk
Wire mark brush electrode surface obtains 2 modified electrodes that different extract liquors have been added dropwise after it is spontaneously dried.It is placed in phosphate buffer
Middle forming circuit obtains two groups of different electrochemical signals after differential pulse voltammetry scans, and constructs the standard two of fresh eupatorium
Tie up electrochemistry finger-print.As a result as shown in Fig. 1.
(2) preparation of medicinal material extract liquor: the puppet of eupatorium known to eupatorium certified products and 4 various names is separately taken to mix product: Herba Lycopi, purple stem pool
Orchid, Herba Serissae, Eupatorium lindleyanum.It respectively takes 0.5g fragment of tissue into 2mL dimethylformamide and methanol solution respectively, sufficiently vibrates
Afterwards, 10 parts of above-mentioned medicinal materials extract liquor in dimethylformamide and methanol respectively is obtained.
(3) electrode modification: respectively taking above-mentioned 2 μ L of extract liquor, is respectively coated with liquid-transfering gun in screen printing electrode surface,
10 modified electrodes that different extract liquors have been added dropwise are obtained after 60 ° of dryings.
(4) electrochemical analysis: 10 sheet modified electrodes are separately placed in phosphate buffer and are formed into a loop, through difference
Two groups of different electrochemical signals are obtained after Pulse Voltammetry scanning.Wherein, the differential pulse voltammetry sweep interval be 1.2V~-
0.2V；Scanning speed is 100mV/s；Sampling interval is 1mV.
Every kind of medicinal material obtains 2 kinds of different electrochemical signals (with each one group of dimethylformamide and methanol extraction), structure
Build the two-dimentional electrochemistry finger-print of each medicinal material.As a result as shown in attached drawing 2~6.
Fig. 2~6 are compared with the standard two-dimensional electrochemistry finger-print of Fig. 1 it is found that the electrochemistry of eupatorium certified products refers to respectively
Line map and standard diagram similarity are up to very high, and electrochemistry finger-print and standard diagram similarity that eupatorium mixes adulterant are very
It is small.Therefore embodiment success identifies eupatorium certified products from fresh eupatorium and its mixed adulterant.
Embodiment 2: the detection to drying herbal medicine
(1) building of eupatorium certified products standard electrochemical finger-print: drying Chinese medicine eupatorium is taken to shred into tissue with scissors
Fragment throws 0.5g sample into 2mL dimethylformamide and methanol solution respectively, sufficiently after oscillation, obtains two parts of drying eupatoriums
Extract liquor in dimethylformamide and methanol respectively.Above-mentioned 2 μ L of extract liquor is respectively taken, is respectively coated with liquid-transfering gun in silk screen
It prints electrode surface, obtains 2 modified electrodes that different extract liquors have been added dropwise after it is spontaneously dried.It is placed in phosphate buffer
Forming circuit, two groups of different electrochemical signals are obtained after differential pulse voltammetry scans, construct the standard two-dimensional of drying eupatorium
Electrochemistry finger-print.As a result as shown in Fig. 7.
(2) preparation of medicinal material extract liquor: drying herbal medicine known to 5 various names: eupatorium, Herba Lycopi, Eupatorium adenophorum, Hua Ze is separately taken
Blue and Eupatorium lindleyanum.It respectively takes 0.5g fragment of tissue into 2mL dimethylformamide and methanol solution respectively, sufficiently after oscillation, obtains
To 10 parts of above-mentioned medicinal materials extract liquor in dimethylformamide and methanol respectively.
(3) electrode modification: above-mentioned 2 μ L of extract liquor is respectively taken, is respectively coated with liquid-transfering gun in screen printing electrode surface, to it
10 modified electrodes that different extract liquors have been added dropwise are obtained after natural drying.
(4) electrochemical analysis: 10 sheet modified electrodes are separately placed in phosphate buffer and are formed into a loop, through difference
Two groups of different electrochemical signals are obtained after Pulse Voltammetry scanning.Wherein, the differential pulse voltammetry sweep interval be 1.2V~-
0.2V；Scanning speed is 100mV/s；Sampling interval is 1mV.
Every kind of medicinal material obtains 2 kinds of different electrochemical signals (with each one group of dimethylformamide and methanol extraction), structure
Build the two-dimentional electrochemistry finger-print of each medicinal material.As a result as shown in attached drawing 8~12.
Fig. 8~12 are compared with the standard two-dimensional electrochemistry finger-print of Fig. 7 it is found that the electrochemistry of eupatorium certified products refers to respectively
Line map and standard diagram similarity are up to very high, and electrochemistry finger-print and standard diagram similarity that eupatorium mixes adulterant are very
It is small.Therefore embodiment success identifies eupatorium certified products from drying eupatorium and its mixed adulterant.
Comparative example 1: fresh eupatorium is detected with normal pulsed voltammetric scan
(1) fresh Chinese medicine eupatorium the building of eupatorium certified products standard electrochemical finger-print: is shredded into tissue with scissors
Fragment, respectively throw 0.5g fragment of tissue into 2mL dimethylformamide and methanol solution, sufficiently oscillation after, obtain two parts it is fresh
The eupatorium extract liquor in dimethylformamide and methanol respectively.Above-mentioned 2 μ L of extract liquor is respectively taken, is respectively coated with liquid-transfering gun in silk
Wire mark brush electrode surface obtains 2 modified electrodes that different extract liquors have been added dropwise after it is spontaneously dried.It is placed in phosphate buffer
Middle forming circuit obtains two groups of different electrochemical signals after normal pulsed voltammetric scan, constructs the standard two of fresh eupatorium
Tie up electrochemistry finger-print.As a result as shown in Fig. 13.
(2) preparation of medicinal material extract liquor: separately take 0.5g eupatorium certified products fragment of tissue molten to 2mL dimethylformamide and methanol
In liquid, sufficiently after oscillation, 2 portions of eupatoriums extract liquor in dimethylformamide and methanol respectively is obtained.
(3) electrode modification: respectively taking above-mentioned 2 μ L of extract liquor, is respectively coated with liquid-transfering gun in screen printing electrode surface,
2 modified electrodes that different extract liquors have been added dropwise are obtained after 60 ° of dryings.
(4) electrochemical analysis: 2 sheet modified electrodes are separately placed in phosphate buffer and are formed into a loop, through common
Two groups of different electrochemical signals are obtained after Pulse Voltammetry scanning.Wherein, normal pulsed voltammetric scan section be 1.2V~-
0.2V；Scanning speed is 100mV/s；Sampling interval is 1mV.
With resulting 2 kinds of different electrochemical signals (with each one group of dimethylformamide and methanol extraction), each medicine is constructed
The two-dimentional electrochemistry finger-print of material.As a result as shown in Fig. 14.
Fig. 1 and 2 in Figure 13 and 14 and embodiment 1 is compared it is found that for same plant, differential pulse is utilized
The otherness for the electrochemistry finger-print that voltammetric scan obtains is smaller than the electrochemistry fingerprint image that normal pulsed voltammetric scan obtains,
Therefore, differential pulse voltammetry scanning possesses higher accuracy in plants identification.
As seen from the above embodiment, the present invention provides one kind based on electrochemical analysis, with eupatorium and its mixed adulterant
For the eupatorium certified products discrimination method for implementing object.With different solvents extraction medicinal material obtain different extracts, by extract modification in
Electrode surface carries out Pulse Voltammetry scanning to the electrode of modified, converts multidimensional electrochemistry figure for obtained electrochemical signals
Spectrum, it is whether consistent by comparing the electrochemistry finger-print and standard electrochemical finger-print of sample, whether judge institute's sample
It is eupatorium certified products.Identification method provided by the invention is simple and easy to do, quickly can filter out eupatorium just from eupatorium and its mixed adulterant
Product.
The method of the present invention that the above embodiments are only used to help understand and its core concept.It should be pointed out that for
For those skilled in the art, without departing from the principle of the present invention, if can also be carried out to the present invention
Dry improvement and modification, these improvement and modification are also fallen into the claims in the present invention protection scope.
Claims (10)
1. a kind of method for identifying the eupatorium true and false, which comprises the steps of:
(1) fragment of tissue of sample to be tested is added separately to extract in n kind different solvents, obtains n kind extract liquor；Wherein
n≥2；
(2) n kind extract liquor is coated on screen printing electrode surface with liquid-transfering gun respectively, n kind modification electricity is obtained after its drying
Pole；
(3) the n kind modified electrode is respectively placed in progress voltammetry scanning in phosphate buffer, respectively by n kind modified electrode
Obtained multi-loop electric chemical signal constructs to obtain the two-dimentional electrochemistry finger-print of sample to be tested；
(4) the electrochemistry finger-print of sample to be tested and the standard two-dimensional electrochemistry finger-print of eupatorium certified products are compared,
If the electrochemistry finger-print of sample to be tested is similar to standard diagram height, sample to be tested is eupatorium；Conversely, being then adulterant.
2. the method according to claim 1, wherein the sample to be tested is the new fresh grass of eupatorium and its mixed adulterant
Medicine or drying herbal medicine.
3. the method according to claim 1, wherein the quality of the fragment of tissue in the step (1) and each molten
The volume ratio of agent is (0.1~1.5) g:(0.5~4) ml.
4. the method according to claim 1, wherein the different solvents in the step (1) include dimethyl methyl
Amide or methanol.
5. the method according to claim 1, wherein coating sample size used is 1~5 μ in the step (2)
l。
6. the method according to claim 1, wherein the drying mode in the step (2) is that 60 DEG C of heating are dried
It is dry.
7. the method according to claim 1, wherein the voltammetric scan mode in the step (3) is difference arteries and veins
Rush voltammetry scanning.
8. the method according to claim 1, wherein the phosphate buffer in the step (3) is biphosphate
The mixed solution of potassium and sodium dihydrogen phosphate, respective concentration are respectively 0.05~0.2mol/L.
9. the method according to claim 1, wherein in the step (3) pulse voltammetric scan sweep interval
For 1.2V~-0.2V.
10. the method according to claim 1, wherein two-dimentional electrochemistry finger-print is by appointing in the step (3)
Anticipate one group of differential pulse voltammetry scanning spectra current value as fingerprint image X-axis data, another component pulse voltammetry scanning
The current value of map is as obtained by fingerprint image Y-axis data.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111272858A (en) * | 2020-04-01 | 2020-06-12 | 杭州电子科技大学 | Analysis method of pottery clay of temple site |
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| CN1403809A (en) * | 2001-09-04 | 2003-03-19 | 洪筱坤 | Establishment and application of Chinese medicine chaacteristic fingerprint spectrum |
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| CN111272858B (en) * | 2020-04-01 | 2022-05-31 | 杭州电子科技大学 | Analysis method of pottery clay of temple site |
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Effective date of registration: 20220217 Address after: 230000 Anhui Hefei high tech Zone Innovation Industrial Park two phase J2 District C block 18 floor. Patentee after: HEFEI LONGTUTEM INFORMATION TECHNOLOGY Co.,Ltd. Address before: 310018 no.1158, No.2 street, Baiyang street, Hangzhou Economic and Technological Development Zone, Zhejiang Province Patentee before: HANGZHOU DIANZI University |