CN109115915A - A kind of detection method of Chinese herbaceous peony plant different parts paeoniflorin content - Google Patents
A kind of detection method of Chinese herbaceous peony plant different parts paeoniflorin content Download PDFInfo
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- CN109115915A CN109115915A CN201811174077.5A CN201811174077A CN109115915A CN 109115915 A CN109115915 A CN 109115915A CN 201811174077 A CN201811174077 A CN 201811174077A CN 109115915 A CN109115915 A CN 109115915A
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
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Abstract
The invention discloses a kind of detection methods of Chinese herbaceous peony plant different parts paeoniflorin content, including drawing materials, separating, weighs, extract Paeoniflorin, filtering, chromatographic isolation, record absorption value, preparation reference substance solution, establish linear relationship, carry out precision test, substitute into linear equation and obtain paeoniflorin content.Not completely crushed bulky grain is screened out, the progress that can ensure to extract at the extraction is consistent, filtering is organic filter membrane using 0.45um, the insoluble substance and bubble of mobile phase and the inside in the Paeoniflorin solution of extraction are filtered, the reference substance solution for choosing 1.0,2.0,4.0,6.0,8.0 and 10.0ml respectively is placed in 10ml measuring bottle, it is configured to gradient concentration, convenient for the formulation of Paeoniflorin regression equation.
Description
Technical field
The present invention relates to a kind of detection method, specially a kind of detection side of Chinese herbaceous peony plant different parts paeoniflorin content
Method belongs to organic compound detection technique field.
Background technique
Chinese herbaceous peony is that Paeoniaceae Paeonia herbaceos perennial has very in the cultivation history that China has more than 4000 years
High ornamental value and medical value, and Paeoniflorin is the main effective of a variety of parts of generic medicinal plants such as Radix Paeoniae Alba, radix paeoniae rubra and cortex moutan
Ingredient has significant ground antalgic and sedative anticonvulsant action, and energy coronary artery dilator increases coronary flow, and confrontation Acute myocardial lacks
Blood reduces blood pressure, protection cerebral ischemia, antithrombus formation, platelet aggregation-against, and lipidemia reverses the effect of low blood pressure, Anhui
Bozhou area is Radix Paeoniae Alba Genuine producing area, high-quality and well-known throughout the country because its yield is big, the harvesting of existing Radix Paeoniae Alba with 3-5 life be it is more,
Some Enterprises are that the Paeoniflorin monomer extracted in Radix Paeoniae Alba is used as raw material.
When extracting Paeoniflorin, often whole plant is extracted for enterprise at present, and contained by Chinese herbaceous peony plant different parts
Paeoniflorin is also different, lacks detection to different parts paeoniflorin content, causes to extract insufficient, causes to waste.
Summary of the invention
Contain the object of the invention is that providing a kind of Chinese herbaceous peony plant different parts Paeoniflorin to solve the above-mentioned problems
The detection method of amount.
The present invention is through the following technical solutions to achieve the above objectives: a kind of Chinese herbaceous peony plant different parts paeoniflorin content
Detection method, detection method include
Step A: materials choose the Chinese herbaceous peony plant of different Growing years；
Step B: separation weighs, and each section flower of Chinese herbaceous peony plant, leaf, stem and root is separated, and weigh identical weight
Component is spare；
Step C: extracting Paeoniflorin, grinding and sieving, and EtOH Sonicate processing 30min is added；
Step D: filtering obtains subsequent filtrate；
Step E: subsequent filtrate is poured into the sample flasket of chromatography by chromatographic isolation, is put into the efficient liquid of Agilent1200 type
In chromatography, chromatogram is obtained；
Step F: record absorption value records numerical value obtained；
Step G: preparation reference substance solution, precision weigh Paeoniflorin reference substance, and methanol is added and is placed in measuring bottle, mixing is made
Solution；
Step H: establishing linear relationship, takes the reference substance solution prepared to configure various concentration, respectively sample introduction 10ul, record
Chromatogram obtains Paeoniflorin regression equation by peak area to corresponding concentration linear regression；
Step I: carrying out precision test, and precision draws reference substance solution 10ul, repeats sample introduction 5 times, measures Paeoniflorin color
Spectral peak area value；
Step J: substituting into linear equation, and the subsequent filtrate absorption value of record is substituted into the Paeoniflorin that reference substance solution obtains and is returned
Equation；
Step K: paeoniflorin content is obtained according to linear equation and calculates separately out different parts paeoniflorin content.
Preferably, in order to reduce error when making detection, the materials in the step A choose three different Growing years
Chinese herbaceous peony plant, and cleaned using distilled water.
Preferably, in order to screen out not completely crushed bulky grain, in the step C, using 40 meshes to smashed
Chinese herbaceous peony each section is sieved.
Preferably, in order to which the insoluble substance and bubble of mobile phase and the inside in the Paeoniflorin solution by extraction carried out
It filters, in the step D, filtering is organic filter membrane using 0.45um.
Preferably, in order to obtain higher separating degree in shorter time, in the step E, the item of chromatographic isolation
Part is chromatographic column Shim-pack vp, and specification: 150x4.6, mobile phase are -0.1% phosphoric acid solution of acetonitrile (14:86), detect wave
Long 230nm, column temperature 22C, sample injector 20u1, theoretical cam curve is calculated by Paeoniflorin peak is not less than 2000.
Preferably, for the ease of the formulation of Paeoniflorin regression equation, in the step H, choose 1.0 respectively, 2.0,4.0,
6.0, the reference substance solution of 8.0 and 10.0ml is placed in 10ml measuring bottle.
The beneficial effects of the present invention are: the detection method design of the Chinese herbaceous peony plant different parts paeoniflorin content is rationally, step
Materials in rapid A, choose the Chinese herbaceous peony plant of three different Growing years, and are cleaned using distilled water, can obtain multiple
Sample is made to reduce error when detection, in step C, is sieved, will not be crushed to smashed Chinese herbaceous peony each section using 40 meshes
Complete bulky grain screens out, and the progress that can ensure to extract at the extraction is consistent, and in step D, filtering is having using 0.45um
The insoluble substance and bubble of mobile phase and the inside in the Paeoniflorin solution of extraction are filtered, prevent them by machine filter film
Block instrument, in step E, the condition of chromatographic isolation is chromatographic column Shim-pack vp, and specification: 150x4.6, mobile phase are second
- 0.1% phosphoric acid solution of nitrile (14:86), Detection wavelength 230nm, column temperature 22C, sample injector 20u1, theoretical cam curve press Paeoniflorin peak
It calculates and is not less than 2000, sensitivity is higher, can choose respectively in the higher separating degree of acquisition of shorter time, step H
1.0, the reference substance solution of 2.0,4.0,6.0,8.0 and 10.0ml is placed in 10ml measuring bottle, is configured to gradient concentration, is convenient for Chinese herbaceous peony
The formulation of glycosides regression equation.
Detailed description of the invention
Fig. 1 is schematic structural view of the invention.
Specific embodiment
Following will be combined with the drawings in the embodiments of the present invention, and technical solution in the embodiment of the present invention carries out clear, complete
Site preparation description, it is clear that described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.It is based on
Embodiment in the present invention, it is obtained by those of ordinary skill in the art without making creative efforts every other
Embodiment shall fall within the protection scope of the present invention.
Referring to Fig. 1, a kind of detection method of Chinese herbaceous peony plant different parts paeoniflorin content, detection method include
Step A: materials choose the Chinese herbaceous peony plant of different Growing years；
Step B: separation weighs, and each section flower of Chinese herbaceous peony plant, leaf, stem and root is separated, and weigh identical weight
Component is spare；
Step C: extracting Paeoniflorin, grinding and sieving, and EtOH Sonicate processing 30min is added；
Step D: filtering obtains subsequent filtrate；
Step E: subsequent filtrate is poured into the sample flasket of chromatography by chromatographic isolation, is put into the efficient liquid of Agilent1200 type
In chromatography, chromatogram is obtained；
Step F: record absorption value records numerical value obtained；
Step G: preparation reference substance solution, precision weigh Paeoniflorin reference substance, and methanol is added and is placed in measuring bottle, mixing is made
Solution；
Step H: establishing linear relationship, takes the reference substance solution prepared to configure various concentration, respectively sample introduction 10ul, record
Chromatogram obtains Paeoniflorin regression equation by peak area to corresponding concentration linear regression；
Step I: carrying out precision test, and precision draws reference substance solution 10ul, repeats sample introduction 5 times, measures Paeoniflorin color
Spectral peak area value；
Step J: substituting into linear equation, and the subsequent filtrate absorption value of record is substituted into the Paeoniflorin that reference substance solution obtains and is returned
Equation；
Step K: paeoniflorin content is obtained according to linear equation and calculates separately out different parts paeoniflorin content.
Materials in the step A are chosen the Chinese herbaceous peony plant of three different Growing years, and are carried out clearly using distilled water
It washes, multiple samples can be obtained, make to reduce error when detection, in the step C, using 40 meshes to each portion of smashed Chinese herbaceous peony
Divide and be sieved, not completely crushed bulky grain is screened out, the progress that can ensure to extract at the extraction is consistent, the step D
In, filtering is organic filter membrane using 0.45um, by the insoluble substance of mobile phase and the inside in the Paeoniflorin solution of extraction
It is filtered with bubble, prevents them from blocking instrument, in the step E, the condition of chromatographic isolation is chromatographic column Shim-pack
Vp, specification: 150x4.6, mobile phase are -0.1% phosphoric acid solution of acetonitrile (14:86), Detection wavelength 230nm, column temperature 22C, sample introduction
Device 20u1, theoretical cam curve is calculated by Paeoniflorin peak is not less than 2000, and sensitivity is higher, can obtain in the shorter time higher
Separating degree, in the step H, the reference substance solution for choosing 1.0,2.0,4.0,6.0,8.0 and 10.0ml respectively is placed in 10ml
In measuring bottle, it is configured to gradient concentration, convenient for the formulation of Paeoniflorin regression equation.
It is obvious to a person skilled in the art that invention is not limited to the details of the above exemplary embodiments, Er Qie
In the case where without departing substantially from spirit or essential attributes of the invention, the present invention can be realized in other specific forms.Therefore, no matter
From the point of view of which point, the present embodiments are to be considered as illustrative and not restrictive, and the scope of the present invention is by appended power
Benefit requires rather than above description limits, it is intended that all by what is fallen within the meaning and scope of the equivalent elements of the claims
Variation is included within the present invention.Any reference signs in the claims should not be construed as limiting the involved claims.
In addition, it should be understood that although this specification is described in terms of embodiments, but not each embodiment is only wrapped
Containing an independent technical solution, this description of the specification is merely for the sake of clarity, and those skilled in the art should
It considers the specification as a whole, the technical solutions in the various embodiments may also be suitably combined, forms those skilled in the art
The other embodiments being understood that.
Claims (6)
1. a kind of detection method of Chinese herbaceous peony plant different parts paeoniflorin content, it is characterised in that: its detection method includes
Step A: materials choose the Chinese herbaceous peony plant of different Growing years；
Step B: separation weighs, and each section flower of Chinese herbaceous peony plant, leaf, stem and root is separated, and weigh the component of identical weight
It is spare；
Step C: extracting Paeoniflorin, grinding and sieving, and EtOH Sonicate processing 30min is added；
Step D: filtering obtains subsequent filtrate；
Step E: subsequent filtrate is poured into the sample flasket of chromatography by chromatographic isolation, is put into Agilent1200 type high-efficient liquid phase color
In spectrometer, chromatogram is obtained；
Step F: record absorption value records numerical value obtained；
Step G: preparation reference substance solution, precision weigh Paeoniflorin reference substance, and methanol is added and is placed in measuring bottle, it is molten that mixing is made
Liquid；
Step H: establishing linear relationship, takes the reference substance solution prepared to configure various concentration, respectively sample introduction 10ul, records chromatography
Figure, by peak area to corresponding concentration linear regression, obtains Paeoniflorin regression equation；
Step I: carrying out precision test, and precision draws reference substance solution 10ul, repeats sample introduction 5 times, measures Paeoniflorin chromatographic peak
Area value；
Step J: substituting into linear equation, and the subsequent filtrate absorption value of record is substituted into the Paeoniflorin recurrence side that reference substance solution obtains
Journey；
Step K: paeoniflorin content is obtained according to linear equation and calculates separately out different parts paeoniflorin content.
2. a kind of detection method of Chinese herbaceous peony plant different parts paeoniflorin content according to claim 1, it is characterised in that:
Materials in the step A, choose the Chinese herbaceous peony plant of three different Growing years, and are cleaned using distilled water.
3. a kind of detection method of Chinese herbaceous peony plant different parts paeoniflorin content according to claim 1, it is characterised in that:
In the step C, it is sieved using 40 meshes to smashed Chinese herbaceous peony each section.
4. a kind of detection method of Chinese herbaceous peony plant different parts paeoniflorin content according to claim 1, it is characterised in that:
In the step D, filtering is organic filter membrane using 0.45um.
5. a kind of detection method of Chinese herbaceous peony plant different parts paeoniflorin content according to claim 1, it is characterised in that:
In the step E, the condition of chromatographic isolation is chromatographic column Shim-pack vp, and specification: 150x4.6, mobile phase are acetonitrile-
0.1% phosphoric acid solution (14:86), Detection wavelength 230nm, column temperature 22C, sample injector 20u1, theoretical cam curve is based on Paeoniflorin peak
It calculates and is not less than 2000.
6. a kind of detection method of Chinese herbaceous peony plant different parts paeoniflorin content according to claim 1, it is characterised in that:
In the step H, the reference substance solution for choosing 1.0,2.0,4.0,6.0,8.0 and 10.0ml respectively is placed in 10ml measuring bottle.
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| US20020160059A1 (en) * | 2000-09-13 | 2002-10-31 | Wei Xiao | Cinnamomi and poria composition, method to prepare same and uses thereof |
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| US20020160059A1 (en) * | 2000-09-13 | 2002-10-31 | Wei Xiao | Cinnamomi and poria composition, method to prepare same and uses thereof |
| CN102106948A (en) * | 2009-12-29 | 2011-06-29 | 郑州豫密药业股份有限公司 | Method for measuring paeoniflorin content in norlutin pills by high performance liquid chromatography |
| CN102998386A (en) * | 2012-11-22 | 2013-03-27 | 宁波立华制药有限公司 | Method for measuring radix paeoniae alba diglucoside in radix paeoniae alba callus by utilizing high performance liquid chromatography |
| CN106841450A (en) * | 2017-02-22 | 2017-06-13 | 黑龙江省科学院自然与生态研究所 | It is a kind of at the same determine Chinese herbaceous peony in 9 kinds of HPLC methods of monomeric compound |
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