CN108508119A - The discrimination method of Rhizoma Phragmitis extract - Google Patents
The discrimination method of Rhizoma Phragmitis extract Download PDFInfo
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- G01N30/02—Column chromatography
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- G01N30/8624—Detection of slopes or peaks; baseline correction
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Abstract
The present invention relates to the field of Chinese medicines, the more particularly to discrimination method of Rhizoma Phragmitis extract.Discrimination method includes the following steps：The characteristic spectrum of Rhizoma Phragmitis extract standard items and reed root extract standard items is obtained respectively；Obtain Rhizoma Phragmitis extract characteristic peak and reed root extract characteristic peak；Sample to be tested is taken to detect, whether discriminating acquisition sample to be tested be in Rhizoma Phragmitis extract and/or the sample to be tested doped with reed root extract according to testing result.It whether can detect in Rhizoma Phragmitis extract mixed with reed root extract by the above method, and specificity is good, high sensitivity, accurate and reliable, when mixing the reed root of 5% or more mass fraction in reed root, the reed root extract in extract can be detected.
Description
Technical field
The present invention relates to the field of Chinese medicines, the more particularly to discrimination method of Rhizoma Phragmitis extract.
Background technology
Chinese medical extract clear, relatively-stationary extract of chemical composition portfolio ratio for base, is consistent
Natural extract.It can substitute crude drug and be used as medicine and ensure the physicochemical characteristic for having safe and stable, be both a kind of extract
It is a kind of new pharmaceutical dosage form, the bulk pharmaceutical chemicals that the standardization at the same time as Chinese patent drug GMP productions feeds intake.
Rhizoma Phragmitis extract is that reed root medicinal material is prepared using suitable technique, and main component is phenolic acid, main
To include p-Coumaric Acid, ferulic acid, caffeic acid etc., have the function of heat-clearing, preventing or arresting vomiting, antibacterial anti-inflammatory, be used for crude drug reed root
The effect for the treatment of fever, preventing or arresting vomiting, matches.
Adulterant of the reed root as reed root, because of the similitude with reed root in title, appearance character and drug effect, some producers are because of profit
Benefit drive or lack profession Chinese medical herb ability, using adulterant medicinal material as raw material or mix raw material in extract.Reed
Root extract and similar to Rhizoma Phragmitis extract character, research find that main component contains p-Coumaric Acid, ferulic acid, but its effect
There is certain difference with Rhizoma Phragmitis extract, is mixed the effect of Rhizoma Phragmitis extract sale can influence Rhizoma Phragmitis extract, seriously invaded
Violate consumers' rights and interests.Since reed root extract and Rhizoma Phragmitis extract have an identical main component, only by p-Coumaric Acid or Ah
Whether Wei's acid content measurement is difficult to discriminate between in Rhizoma Phragmitis extract mixed with reed root extract.
Invention content
In view of this, the present invention provides a kind of discrimination method of Rhizoma Phragmitis extract.This method can detect in Rhizoma Phragmitis extract
Whether mixed with reed root extract, and specificity is good, high sensitivity, accurate and reliable, when mixed in reed root mass fraction 5% and with
On reed root when, the reed root extract in extract can be detected.
In order to achieve the above-mentioned object of the invention, the present invention provides following technical scheme：
The present invention provides a kind of discrimination methods of Rhizoma Phragmitis extract, include the following steps：
Step 1：The characteristic spectrum of Rhizoma Phragmitis extract standard items and reed root extract standard items is obtained respectively；
Step 2：Obtain Rhizoma Phragmitis extract characteristic peak and reed root extract characteristic peak；
Step 3：It takes sample to be tested to detect, is differentiated according to the testing result and obtain whether the sample to be tested is that reed root carries
It whether takes in object and/or the sample to be tested doped with reed root extract.
In some specific embodiments of the present invention, the method for the detection is high performance liquid chromatography；
Its chromatographic condition is：Stationary phase is octadecylsilane chemically bonded silica column；It is eluted using flow gradient, mobile phase A
For acetonitrile, Mobile phase B is 0.5% potassium dihydrogen phosphate (pH=2.15), A:B=7%~22%：93%~78%；Flow velocity
0.8~1.2mL/min；Detection wavelength is 270~280nm；Chromatogram column temperature is 25~35 DEG C.
In some specific embodiments of the present invention, the chromatographic condition is specially：A, B phase transformation of eluent turn to：0
~15min, A phase 7%；15~35min, A phase 7%~12%；35~55min, A phase 12%~15%；55~70min, A phase
15%~19%；70~80min, A phase 19%~22%；80~85min, A phase 22%；Flow velocity 1.0mL/min；Detection wavelength is
275nm；Chromatogram column temperature is 30 DEG C.
In some specific embodiments of the present invention, the Rhizoma Phragmitis extract characteristic peak totally 7, retention time range
For 15~50min；The reed root extract characteristic peak totally 6, retention time ranging from 55~85min.
In some specific embodiments of the present invention, the Rhizoma Phragmitis extract characteristic peak totally 7, retention time is
17.1 ± 2min, 23.8 ± 2min, 26.7 ± 2min, 28.3 ± 2min, 35.1 ± 2min, 39.2 ± 2min and 46.6 ±
2min；The reed root extract characteristic peak totally 6, retention time be 59.6 ± 2min, 65.0 ± 2min, 71.5 ± 2min,
75.0 ± 2min, 78.4 ± 2min and 80.0 ± 2min.
In some specific embodiments of the present invention, the standard of the discriminating is：
I) if, the testing result include that all the Rhizoma Phragmitis extract characteristic peak, the sample to be tested are reed
Root extract；, whereas if only including the part Rhizoma Phragmitis extract characteristic peak, the then sample to be tested in the testing result
It is not Rhizoma Phragmitis extract；
If II), the sample to be tested includes whole Rhizoma Phragmitis extract characteristic peaks, and including at least described in one
Reed root extract characteristic peak, then the sample to be tested is for Rhizoma Phragmitis extract and doped with reed root extract；, whereas if described wait for
Sample includes whole Rhizoma Phragmitis extract characteristic peaks, but is not reed root extract characteristic peak including described in, then described to wait for
Sample is Rhizoma Phragmitis extract and undopes and have reed root extract.
In some specific embodiments of the present invention, the retention time of the Rhizoma Phragmitis extract characteristic peak is 15~
50min；The retention time of the reed root extract characteristic peak is 55~85min.
In some specific embodiments of the present invention, the retention time of the Rhizoma Phragmitis extract characteristic peak is 17.1 ±
2min, 23.8 ± 2min, 26.7 ± 2min, 28.3 ± 2min, 35.1 ± 2min, 39.2 ± 2min and 46.6 ± 2min；It is described
The retention time of reed root extract characteristic peak is 59.6 ± 2min, 65.0 ± 2min, 71.5 ± 2min, 75.0 ± 2min, 78.4
± 2min and 80.0 ± 2min.
The present invention some specific embodiments in, in the sample to be tested detection of reed root extract be limited to be not less than
5% (w/w).
The present invention provides a kind of discrimination methods of Rhizoma Phragmitis extract, include the following steps：Step 1：Reed root is obtained respectively
The characteristic spectrum of extract standard items and reed root extract standard items；Step 2：Obtain Rhizoma Phragmitis extract characteristic peak and reed root extraction
Object characteristic peak；Step 3：It takes sample to be tested to detect, is differentiated according to the testing result and obtain whether the sample to be tested is reed root
Whether doped with reed root extract in extract and/or the sample to be tested.Can detect in Rhizoma Phragmitis extract by the above method is
No and specificity is good, high sensitivity, accurate and reliable mixed with reed root extract, when mixing 5% or more mass fraction in reed root
Reed root when, the reed root extract in extract can be detected.
Description of the drawings
In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show below
There is attached drawing needed in technology description to be briefly described.
Fig. 1 shows precision test overlay chart；
Fig. 2 shows stability test overlay chart；
Fig. 3 shows repetitive test overlay chart；
Fig. 4 shows the extract characteristic spectrum overlay chart of different batches reed root；
Fig. 5 shows the compare feature collection of illustrative plates that the extract of different batches reed root generates；
Fig. 6 shows the extract characteristic spectrum overlay chart of different batches reed root；
Fig. 7 shows the compare feature collection of illustrative plates that the extract of different batches reed root generates；
Fig. 8 shows the Rhizoma Phragmitis extract HPLC chromatogram of 5% reed root of incorporation；
Fig. 9 shows the Rhizoma Phragmitis extract HPLC chromatogram of 10% reed root of incorporation；
Figure 10 shows the Rhizoma Phragmitis extract HPLC chromatogram of 15% reed root of incorporation；
Figure 11 shows the Rhizoma Phragmitis extract HPLC chromatogram of 20% reed root of incorporation；
Figure 12 shows the HPLC chromatogram of not adulterated Rhizoma Phragmitis extract；
Figure 13 shows Rhizoma Phragmitis extract (LG1) characteristic peak ultraviolet spectra；
Figure 14 shows 6 characteristic peak (w1~w6) ultraviolet spectras of reed root extract (WG1-ZZ).
Specific implementation mode
The invention discloses a kind of discrimination method of Rhizoma Phragmitis extract, those skilled in the art can use for reference present disclosure,
It is suitably modified technological parameter realization.In particular, it should be pointed out that all similar substitutions and modifications carry out those skilled in the art
Say it is it will be apparent that they are considered as being included in the present invention.The method of the present invention and application have passed through preferred embodiment
It is described, related personnel can obviously not depart from the content of present invention, in spirit and scope to method described herein and answer
With being modified or suitably changing and combine, to realize and apply the technology of the present invention.
The purpose of the present invention is to provide a kind of Rhizoma Phragmitis extract distinguishing method between true and false, are for differentiating in Rhizoma Phragmitis extract
It is no mixed with reed root extract.
The technical solution used in the present invention is：
A kind of Rhizoma Phragmitis extract distinguishing method between true and false, includes the following steps：
1) high performance liquid chromatography detection Rhizoma Phragmitis extract standard items and reed root extract standard items are respectively adopted, establish reed
Root extract characteristic spectrum and reed root extract characteristic spectrum；
2) reed root extract characteristic spectrum and Rhizoma Phragmitis extract characteristic spectrum are compared, determine Rhizoma Phragmitis extract characteristic peak and
Reed root extract characteristic peak；
3) use high performance liquid chromatography to detect Rhizoma Phragmitis extract, by measurement result judge in Rhizoma Phragmitis extract whether mixed with
Reed root extract.
Further, high performance liquid chromatography includes the following steps：
1) preparation of reference substance solution：Reference substance p-Coumaric Acid and ferulic acid are taken, is set in brown measuring bottle, methanol is added, is shaken up；
2) preparation of test solution：Precision weighs detection sample, methanol is added, ultrasonic extraction is cooling, filtration, filtrate
It is evaporated, residue adds methanol to dissolve, and is transferred in measuring bottle, shakes up to get test solution；
3) it measures：Precision draws test solution, and injection high performance liquid chromatograph measures.
Further, 22.2 μ g/mL p-Coumaric Acids and 10.6 μ g/mL ferulic acids are contained in reference substance solution.
Further, chromatographic condition when high performance liquid chromatography detection is：Stationary phase is octadecylsilane bonded silica
Rubber column gel column；It is eluted using flow gradient, mobile phase A is acetonitrile, and Mobile phase B is 0.5% potassium dihydrogen phosphate (pH=2.15), A:
B=7%~22%：93%~78%；0.8~1.2mL/min of flow velocity；Detection wavelength is 270~280nm；Chromatogram column temperature is
25~35 DEG C.
Further, A, B phase transformation of eluent turn to：0~15min, A phase 7%；15~35min, A phase 7%~12%；
35~55min, A phase 12%~15%；55~70min, A phase 15%~19%；70~80min, A phase 19%~22%；80~
85min, A phase 22%；Flow velocity 1.0mL/min；Detection wavelength is 275nm；Chromatogram column temperature is 30 DEG C.
Further, Rhizoma Phragmitis extract characteristic peak totally 7, retention time ranging from 15~50min；Reed root extract is special
Levy totally 6, peak, retention time ranging from 55~85min.
Further, Rhizoma Phragmitis extract characteristic peak totally 7, retention time are 17.1 ± 2min, 23.8 ± 2min, 26.7
± 2min, 28.3 ± 2min, 35.1 ± 2min, 39.2 ± 2min and 46.6 ± 2min；Reed root extract characteristic peak totally 6,
Retention time is 59.6 ± 2min, 65.0 ± 2min, 71.5 ± 2min, 75.0 ± 2min, 78.4 ± 2min and 80.0 ± 2min.
Further, judge be mixed with the standard of reed root extract in Rhizoma Phragmitis extract：Occur reed root in measurement result to carry
Whole characteristic peaks of object are taken, and the characteristic peak of a reed root extract at least occur, illustrate to carry mixed with reed root in Rhizoma Phragmitis extract
Object is taken, otherwise is illustrated in Rhizoma Phragmitis extract not mixed with reed root extract.
Further, judge be mixed with the standard of reed root extract in Rhizoma Phragmitis extract：Measurement result is in retention time 15
There are whole characteristic peaks of Rhizoma Phragmitis extract in~50min, and reed root extract at least occurs in 55~85min of retention time
Characteristic peak illustrates mixed with reed root extract in Rhizoma Phragmitis extract, otherwise illustrates in Rhizoma Phragmitis extract not mixed with reed root extract.
Further, judge be mixed with the standard of reed root extract in Rhizoma Phragmitis extract：Measurement result is in retention time
17.1 ± 2min, 23.8 ± 2min, 26.7 ± 2min, 28.3 ± 2min, 35.1 ± 2min, 39.2 ± 2min and 46.6 ± 2min
Equal appearance, and retention time be 59.6 ± 2min, 65.0 ± 2min, 71.5 ± 2min, 75.0 ± 2min, 78.4 ± 2min and
80.0 ± 2min at least goes out a peak, illustrates mixed with reed root extract in Rhizoma Phragmitis extract, otherwise illustrates in Rhizoma Phragmitis extract not
Mixed with reed root extract.
The beneficial effects of the invention are as follows：It whether can detect in Rhizoma Phragmitis extract mixed with reed root extract by the above method,
And it is the good, high sensitivity of specificity, accurate and reliable, when mixing the reed root of 5% or more mass fraction in reed root, extract
In reed root extract can be detected.
Raw materials used, auxiliary material and reagent are available on the market in the discrimination method of Rhizoma Phragmitis extract provided by the invention.
With reference to embodiment, the present invention is further explained：
Embodiment 1
Instrument and reagent：
Instrument：The efficient high performance liquid chromatograph of Agilent1200 types (diode array detector), the accurate electricity of BP-211D
Sub- balance (Sartorius AG, Goettingen, Germany)；(Switzerland Mettler Toledo are public for MS 204S electronic balances
Department) desk-top ultrasonic cleaning machine (SK7200LH, Shanghai High Kudos Science Instrument Co., Ltd.).
Reagent：7 batches of Rhizoma Phragmitis extracts are commercially available Rhizoma Phragmitis extract, are all made of water extraction extraction.7 batches of reed root extracts are by reality
The homemade extract of reed root medicinal material that separate sources is collected in room is tested, specifically adds 20 times of amount water to decoct three times by appropriate medicinal material, merges
Filtrate is concentrated into thick paste, adds appropriate amount of auxiliary materials mixing, is dried under reduced pressure and obtains.P-Coumaric Acid reference substance (lot number BCBS872), is purchased from
Sigma Aldriches, purity >=98.0%；Ferulic acid reference substance (lot number 111073-201604) is purchased from Chinese food medicine
Research institute, purity 99.0% are determined in product examine；Extraction is that analysis is pure with methanol；Liquid phase analysis reagent acetonitrile is chromatographically pure.
High performance liquid chromatography step：
1) preparation of reference solution：Take p-Coumaric Acid reference substance, ferulic acid reference substance appropriate, it is accurately weighed, set brown
In measuring bottle, add 50% methanol that mixed solutions of every 1mL containing 22.2 μ g of p-Coumaric Acid, 10.6 μ g of ferulic acid is made, shake up to get.
2) preparation of test solution：Precision weighs sample to be tested 0.5g, and 50% methanol 50mL, ultrasonic extraction is added
60min takes out, lets cool, and filters, and filtrate is evaporated, and residue adds 50% methanol to dissolve, and is transferred in 5mL measuring bottles, shakes up, is configured to
The test solution of 0.1g/mL to get.
3) it measures：Accurate respectively to draw 15 μ L reference solutions and test solution, injection high performance liquid chromatograph measures.
Chromatographic condition is：
Use octadecylsilane chemically bonded silica for stationary phase, column model be Zorbax SB C18 (250mm ×
4.6mm, 5 μm)；Mobile phase A is acetonitrile, and Mobile phase B is 0.5% potassium dihydrogen phosphate (pH=2.15)；Flow gradient elutes
In the process, A, B phase transformation of eluent turn to：0~15min, A phase 7%；15~35min, A phase 7%~12%；35~55min, A
Phase 12%~15%；55~70min, A phase 15%~19%；70~80min, A phase 19%~22%；80~85min, A phase
22%；Detection wavelength is 275nm；Flow velocity 1.0mL/min；Chromatogram column temperature is 30 DEG C.
Precision Experiment
It takes with a Rhizoma Phragmitis extract sample test solution, by above-mentioned chromatographic condition continuous sample introduction 6 times, measurement result is such as
Shown in Fig. 1, S1-S6 is the collection of illustrative plates of continuous 6 sample introductions, the relative retention time at each shared peak and opposite peak known to interpretation of result
The RSD of area is respectively less than 3%, shows that precision is good.
Stability experiment
Take with a Rhizoma Phragmitis extract sample test solution, by above-mentioned chromatographic condition respectively 0,3,6,12,18, for 24 hours
Sample introduction measures, and measurement result is shown in Fig. 2, S1-S6 0,3,6,12,18, for 24 hours sample introduction measure as a result, by interpretation of result it is found that respectively
The relative retention time at shared peak and the RSD of relative peak area are respectively less than 3%, show that sample solution is stablized interior for 24 hours.
Repeated experiment
Same a collection of 6 parts of Rhizoma Phragmitis extract sample is taken, it is accurately weighed, it is molten to prepare test sample by above-mentioned test sample preparation method
Liquid, difference sample detection, the results are shown in Figure 3, and S1-S6 is that 6 sample introductions measure as a result, by interpretation of result it is found that each shared
The relative retention time at peak and the RSD of relative peak area are respectively less than 3%, show repeated preferable.
The foundation of characteristic spectrum：
1. the foundation of Rhizoma Phragmitis extract characteristic spectrum
The extract sample for taking different batches reed root is measured according to above-mentioned high performance liquid chromatography step with condition, is surveyed
Fixed the results are shown in Figure 4, and it is as shown in Figure 5 to formulate Rhizoma Phragmitis extract compare feature collection of illustrative plates according to measurement result.
2. the foundation of reed root extract characteristic spectrum：
The extract sample for taking different batches reed root is measured according to above-mentioned high performance liquid chromatography step with condition, root
The results are shown in Figure 6 according to surveying and determination, and it is as shown in Figure 7 to formulate reed root extract compare feature collection of illustrative plates according to measurement result.
The determination of characteristic peak：
By comparing Rhizoma Phragmitis extract characteristic spectrum and reed root extract characteristic spectrum, determine 59.6 ± 2min, 65.0 ±
6 chromatographic peaks that 2min, 71.5 ± 2min, 75.0 ± 2min, 78.4 ± 2min, 80.0 ± 2min occur are that reed root extract is special
Have, and Rhizoma Phragmitis extract occurs in this retention time without chromatographic peak, therefore, this 6 chromatographic peaks is determined as characteristic peak.
Embodiment 2
Instrument and reagent：
Instrument：The efficient high performance liquid chromatograph of Agilent1200 types (diode array detector), the accurate electricity of BP-211D
Sub- balance (Sartorius AG, Goettingen, Germany)；(Switzerland Mettler Toledo are public for MS 204S electronic balances
Department) desk-top ultrasonic cleaning machine (SK7200LH, Shanghai High Kudos Science Instrument Co., Ltd.).
Reagent：7 batches of Rhizoma Phragmitis extracts are commercially available Rhizoma Phragmitis extract, are all made of water extraction extraction.7 batches of reed root extracts are by reality
The homemade extract of reed root medicinal material that separate sources is collected in room is tested, specifically adds 20 times of amount water to decoct three times by appropriate medicinal material, merges
Filtrate is concentrated into thick paste, adds appropriate amount of auxiliary materials mixing, is dried under reduced pressure and obtains.P-Coumaric Acid reference substance (lot number BCBS872), is purchased from
Sigma Aldriches, purity >=98.0%；Ferulic acid reference substance (lot number 111073-201604) is purchased from Chinese food medicine
Research institute, purity 99.0% are determined in product examine；Extraction is that analysis is pure with methanol；Liquid phase analysis reagent acetonitrile is chromatographically pure.
High performance liquid chromatography step：
4) preparation of reference solution：Take p-Coumaric Acid reference substance, ferulic acid reference substance appropriate, it is accurately weighed, set brown
In measuring bottle, add 50% methanol that mixed solutions of every 1mL containing 22.2 μ g of p-Coumaric Acid, 10.6 μ g of ferulic acid is made, shake up to get.
5) preparation of test solution：Precision weighs sample to be tested 0.5g, and 50% methanol 50mL, ultrasonic extraction is added
60min takes out, lets cool, and filters, and filtrate is evaporated, and residue adds 50% methanol to dissolve, and is transferred in 5mL measuring bottles, shakes up, is configured to
The test solution of 0.1g/mL to get.
6) it measures：Accurate respectively to draw 15 μ L reference solutions and test solution, injection high performance liquid chromatograph measures.
Chromatographic condition is：
Use octadecylsilane chemically bonded silica for stationary phase, column model be Zorbax SB C18 (250mm ×
4.6mm, 5 μm)；Mobile phase A is acetonitrile, and Mobile phase B is 0.5% potassium dihydrogen phosphate (pH=2.15)；Flow gradient elutes
In the process, A, B phase transformation of eluent turn to：0~15min, A phase 7%；15~35min, A phase 7%~12%；35~55min, A
Phase 12%~15%；55~70min, A phase 15%~19%；70~80min, A phase 19%~22%；80~85min, A phase
22%；Detection wavelength is 275nm；Flow velocity 1.0mL/min；Chromatogram column temperature is 30 DEG C.
Accuracy, sensitivity experiment
The component of sample to be tested A, B, C, D are as follows：
Table 1
| Sample name | A | B | C | D | Control group |
| Reed root | 47.5g | 45g | 42.5g | 40g | 50g |
| Reed root | 2.5g | 5g | 7.5g | 10g | 0 |
| Adulterated amount | 5% | 10% | 15% | 20% | 0 |
Sample A, B, C, D and control sample are taken, water extraction prepares extract A, B, C, D, according to above-mentioned high-efficient liquid phase color
Spectrum step is measured extract with experiment condition, and measurement result is shown in that Figure 12 is seen in Fig. 8~11, control group measurement result.By tying
Fruit it is found that incorporation reed root extract chromatogram 59.6 ± 2min, 65.0 ± 2min, 71.5 ± 2min, 75.0 ± 2min,
There is at least one apparent chromatographic peak in 78.4 ± 2min, 80.0 ± 2min or so, according to identification method provided by the invention it is found that
Mixed with reed root in sample A, B, C, D.
And the extract for being not added with reed root shows in control sample herein without apparent chromatographic peak not mixed with reed root.
And in figure the result shows that, characteristic peak increases, the side in showing through the invention with the increase of reed root adding proportion
Method can differentiate that the Rhizoma Phragmitis extract of incorporation reed root, accuracy are good；And it can differentiate that content is not less than the reed root of 5% (w/w), spirit
Sensitivity is high.
The 3 Rhizoma Phragmitis extract true and false of embodiment differentiates specificity analysis
The foundation and analysis of Rhizoma Phragmitis extract characteristic spectrum：
Under the conditions of 275nm Detection wavelengths, 7 batches of Rhizoma Phragmitis extract sample principal character peaks are almost the same, before and after matching
Overlapping see Fig. 4, using similarity software generate control collection of illustrative plates, indicate 7 characteristic peaks altogether, wherein authenticated p-Coumaric Acid,
Two characteristic peaks of ferulic acid, are shown in Fig. 5.The characteristic peak ultraviolet spectra of 7 characteristic peaks of Rhizoma Phragmitis extract 1 (LG1) is shown in Figure 13.With peak
6 (p-Coumaric Acids) calculate each characteristic peak relative retention time and relative peak area, the results are shown in Table 2 as with reference to peak.
27 batches of Rhizoma Phragmitis extract HPLC characteristic spectrums of table share the relative retention time and relative peak area at peak
The foundation and analysis of reed root extract characteristic spectrum：
Under the conditions of 275nm Detection wavelengths, 7 batches of reed root extract samples can detect 7 spies consistent with Rhizoma Phragmitis extract
Levy peak, but more than Rhizoma Phragmitis extract 6 flavones ultraviolet spectrum characteristics characteristic peak (peak w1-w6), discriminating purpose can be reached.It is different
Batch principal character peak is almost the same, matches front and back overlapping and sees Fig. 6.By the similarity software generation pair of 7 batches or more samples
According to collection of illustrative plates, Fig. 7 is seen.The characteristic peak ultraviolet spectra of 6 flavones characteristic peaks of reed root extract sample WG1-ZZ is shown in Figure 14.With peak 6
(p-Coumaric Acid) calculates each characteristic peak relative retention time and relative peak area, the results are shown in Table 3 as with reference to peak.
37 batches of reed root extract characteristic peak relative retention times of table and relative peak area
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Claims (9)
1. a kind of discrimination method of Rhizoma Phragmitis extract, which is characterized in that include the following steps：
Step 1：The characteristic spectrum of Rhizoma Phragmitis extract standard items and reed root extract standard items is obtained respectively；
Step 2：Obtain Rhizoma Phragmitis extract characteristic peak and reed root extract characteristic peak；
Step 3：It takes sample to be tested to detect, is differentiated according to the testing result and obtain whether the sample to be tested is Rhizoma Phragmitis extract
And/or whether doped with reed root extract in the sample to be tested.
2. discrimination method according to claim 1, which is characterized in that the method for the detection is high performance liquid chromatography；
Its chromatographic condition is：Stationary phase is octadecylsilane chemically bonded silica column；It is eluted using flow gradient, mobile phase A is second
Nitrile, Mobile phase B are 0.5% potassium dihydrogen phosphate (pH=2.15), A:B=7%~22%：93%~78%；Flow velocity 0.8~
1.2mL/min；Detection wavelength is 270~280nm；Chromatogram column temperature is 25~35 DEG C.
3. discrimination method according to claim 2, which is characterized in that the chromatographic condition is specially：A, B phase of eluent
Variation is：0~15min, A phase 7%；15~35min, A phase 7%~12%；35~55min, A phase 12%~15%；55~
70min, A phase 15%~19%；70~80min, A phase 19%~22%；80~85min, A phase 22%；Flow velocity 1.0mL/min；
Detection wavelength is 275nm；Chromatogram column temperature is 30 DEG C.
4. discrimination method according to any one of claims 1 to 3, which is characterized in that the Rhizoma Phragmitis extract characteristic peak totally 7
It is a, retention time ranging from 15~50min；The reed root extract characteristic peak totally 6, retention time ranging from 55~
85min。
5. discrimination method according to any one of claims 1 to 4, which is characterized in that the Rhizoma Phragmitis extract characteristic peak totally 7
It is a, retention time be 17.1 ± 2min, 23.8 ± 2min, 26.7 ± 2min, 28.3 ± 2min, 35.1 ± 2min, 39.2 ±
2min and 46.6 ± 2min；The reed root extract characteristic peak totally 6, retention time be 59.6 ± 2min, 65.0 ± 2min,
71.5 ± 2min, 75.0 ± 2min, 78.4 ± 2min and 80.0 ± 2min.
6. discrimination method according to any one of claims 1 to 5, which is characterized in that the standard of the discriminating is：
I) if, the testing result include that all the Rhizoma Phragmitis extract characteristic peak, the sample to be tested carry for reed root
Take object；, whereas if only including the part Rhizoma Phragmitis extract characteristic peak in the testing result, then the sample to be tested is not
Rhizoma Phragmitis extract；
If II), the sample to be tested includes whole Rhizoma Phragmitis extract characteristic peaks, and includes at least a reed root
Extract characteristic peak, then the sample to be tested is for Rhizoma Phragmitis extract and doped with reed root extract；, whereas if described wait for test sample
Product include whole Rhizoma Phragmitis extract characteristic peaks, but do not include the reed root extract characteristic peak, then the sample to be tested
For Rhizoma Phragmitis extract and undopes and have reed root extract.
7. discrimination method according to claim 6, which is characterized in that the retention time of the Rhizoma Phragmitis extract characteristic peak is
15~50min；The retention time of the reed root extract characteristic peak is 55~85min.
8. discrimination method according to claim 6, which is characterized in that the retention time of the Rhizoma Phragmitis extract characteristic peak is
17.1 ± 2min, 23.8 ± 2min, 26.7 ± 2min, 28.3 ± 2min, 35.1 ± 2min, 39.2 ± 2min and 46.6 ±
2min；The retention time of the reed root extract characteristic peak be 59.6 ± 2min, 65.0 ± 2min, 71.5 ± 2min, 75.0 ±
2min, 78.4 ± 2min and 80.0 ± 2min.
9. according to claim 1 to 8 any one of them discrimination method, which is characterized in that reed root is extracted in the sample to be tested
The detection of object is limited to be not less than 5% (w/w).
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