CN106841501A - Simple and fast differentiates the thin-layered chromatography of Radix Glycyrrhizae, swollen fruit Radix and glycyrrhiza glabra - Google Patents
Simple and fast differentiates the thin-layered chromatography of Radix Glycyrrhizae, swollen fruit Radix and glycyrrhiza glabra Download PDFInfo
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Abstract
The present invention relates to the thin-layered chromatography that simple and fast differentiates Radix Glycyrrhizae, swollen fruit Radix and glycyrrhiza glabra.Specifically, the inventive method is comprised the following steps：Extracting liquorice control medicinal material, swollen fruit Radix control medicinal material, plus methyl alcohol or ethanol solution are made control medicinal material solution；Extracting liquorice, swollen fruit Radix, three kinds of medicinal material coarse powders of glycyrrhiza glabra, solubilizer are ultrasonically treated, and Radix Glycyrrhizae, swollen fruit Radix, the need testing solution of glycyrrhiza glabra are obtained respectively；Draw control medicinal material solution and appropriate need testing solution respectively with quantitative capillary tube, point sample is in reversed phase thin layer plate；With 0.1% phosphate aqueous solution and ethanol with certain proportion as solvent, launched, taken out after expansion, dried；5) spot compared in chromatogram differentiates Radix Glycyrrhizae, swollen fruit Radix, glycyrrhiza glabra.Three kinds of source Radix Glycyrrhizaes are presented dramatically different spot in the inventive method gained chromatogram.Discrimination method specificity of the present invention is strong, can accurately differentiate Radix Glycyrrhizae, swollen fruit Radix, glycyrrhiza glabra.
Description
Technical field
The invention belongs to technical field of traditional Chinese medicines, specifically Chinese medicinal material authentication technique field, and in particular to one kind for it is simple,
Rapidly differentiate Radix Glycyrrhizae, swollen fruit Radix, the inverse thin layer chromatography method of glycyrrhiza glabra.
Background technology
Radix Glycyrrhizae is conventional Chinese medicine, first recorded in《Sheng Nong's herbal classic》, it is classified as top grade.It is clearing heat and detoxicating with invigorating the spleen and benefiting qi, moistening lung
Cough-relieving, relieving spasm to stop pain, effect of coordinating the drug actions of a prescription.Radix Glycyrrhizae has turned into the maximum herbal species of drug matching consumption.Version in 2015《In
State's pharmacopeia》Specify in one Radix Glycyrrhizae kind recorded of page 86, Radix Glycyrrhizae is glycyrrhizic legume Glycyrrhiza uralensis
Fish., the dry root of swollen fruit Radix Glycyrrhiza inflate Bat. or glycyrrhiza glabra Glycyrrhiza glabra L.
And rhizome.Made a distinction for ease of the Radix Glycyrrhizae with swollen fruit Radix, two kinds of sources of glycyrrhiza glabra, according to Radix Glycyrrhizae Glycyrrhiza
The latin name epitheton uralensis of uralensis Fish, normally referred to as Glycyrrhiza Uralensis.
According to height dawn beautiful grade, [Gao Xiaojuan, Zhao Dan, Zhao Jianjun wait herbal textual [J] Chinese experimental pharmacologies of traditional Chinese medical formulae of Radix Glycyrrhizaes miscellaneous
Will, 2017,23 (2):193-198] to investigate, China's history tree records Radix Glycyrrhizae and is Glycyrrhiza Uralensis (i.e. 2015 versions《China
Pharmacopeia》One Radix Glycyrrhizae Glycyrrhiza uralensis Fish. of regulation), do not include《Chinese Pharmacopoeia》The swollen fruit recorded is sweet
Careless (Glycyrrhiza inflate Bat.) and glycyrrhiza glabra (Glycyrrhiza glabra L.).
From going through version《Chinese Pharmacopoeia》Situation is recorded as can be seen that version in 1963《Chinese Pharmacopoeia》Only specify the former plant of Radix Glycyrrhizae
Originate is Radix Glycyrrhizae Glycyrrhiza uralensis Fish., version in 1977《Chinese Pharmacopoeia》Specify the former plant origin of Radix Glycyrrhizae
Increased two kinds of plants of swollen fruit Radix Glycyrrhiza inflate Bat. and glycyrrhiza glabra Glycyrrhiza glabra L.
Thing, later each version《Chinese Pharmacopoeia》Continue to use version regulation in 1977.
On the one hand, due to from Radix Glycyrrhizae Glycyrrhiza uralensis Fish., swollen fruit Radix Glycyrrhiza
Chemical composition contained by the licorice medicinal materials of inflate Bat. or glycyrrhiza glabra Glycyrrhiza glabra L. is different, separately
On the one hand, ancient Chinese medicine doctor is using only a kind of Radix Glycyrrhizae Glycyrrhiza uralensis licorice medicinal materials in source of Fish., and three kinds
The licorice medicinal materials appearance character in source is not easy to distinguish.
Although technically, can realize that three kinds of sources are sweet by sophisticated analytical instruments such as high performance liquid chromatography etc.
The discriminating of grass is distinguished, but has not yet to see method simple and easy to apply while realizing that the discriminating of three kinds of source Radix Glycyrrhizaes is distinguished.And
This realistic problem is also that those skilled in the art urgently expect to solve.
The content of the invention
To differentiate simultaneously that three kinds of Radix Glycyrrhizaes in source, swollen fruits are sweet it is an object of the invention to provide a kind of reverse phase thin layer chromatography
Grass, the method for glycyrrhiza glabra, expect source and difference medication that this method is easy to user to differentiate licorice medicinal materials, while expecting
This method is easy, easy to operate, and exactly can make a distinction the Radix Glycyrrhizae in three kinds of sources.It has been found by the present inventors that with
Radix Glycyrrhizae control medicinal material that National Institute for Food and Drugs Control issues, swollen fruit Radix control medicinal material are index components, by solvent
The program of expansion, realizes the discriminating of Radix Glycyrrhizae, swollen fruit Radix, glycyrrhiza glabra, reaches the accurate purpose using medicinal material.
In the present invention, if not otherwise specified, the Radix Glycyrrhizae for referring to, refers to glycyrrhizic legume Glycyrrhiza
Uralensis Fish., i.e. people usually said Glycyrrhiza Uralensis.
The technical solution adopted by the present invention is：
The reverse phase thin layer chromatography of technical scheme 1. differentiates Radix Glycyrrhizae, swollen fruit Radix, the method for glycyrrhiza glabra, it is characterised in that bag
Include following steps：
1) Radix Glycyrrhizae control medicinal material, swollen fruit Radix control medicinal material are weighed respectively, plus methyl alcohol or ethanol solution are made control medicinal material
Solution；
2) extracting liquorice, swollen fruit Radix, three kinds of medicinal material coarse powders of glycyrrhiza glabra are appropriate, are respectively placed in conical flask with cover, solubilization
Agent, ultrasonically treated (such as 5~60 minutes), filtration takes subsequent filtrate, and the confession examination of Radix Glycyrrhizae, swollen fruit Radix, glycyrrhiza glabra is obtained respectively
Product solution；
3) with quantitative capillary tube distinguish aspiration step 1) obtained in two kinds of control medicinal material solution and step 2) obtained in three kinds
Appropriate need testing solution, point sample is on the same horizontal line diverse location of same reversed phase thin layer plate successively；
4) with 0.1% phosphate aqueous solution and ethanol with certain proportion as solvent, launched, taken out after expansion, dried；
5) spot compared as follows in chromatogram differentiates Radix Glycyrrhizae, swollen fruit Radix, glycyrrhiza glabra：
In reviewing step 4 under 365nm length ultraviolet lamps) gained reversed phase thin layer plate, and compare the fluorescence spot in chromatogram,
To differentiate Radix Glycyrrhizae, swollen fruit Radix, glycyrrhiza glabra；
Radix Glycyrrhizae test sample and Radix Glycyrrhizae control medicinal material have a sapphire fluorescent spot at the place of Rf value Rf values 0.41 or so
Point, at the place of Rf values 0.51 or so, there is a fluorescence spot for yellow green；
Swollen fruit Radix test sample and swollen fruit Radix control medicinal material only at the place of Rf values 0.41 or so, have one it is sapphire glimmering
Light spot；
Glycyrrhiza glabra test sample respectively has a fluorescence spot for yellow green, in Rf values at the place of Rf values 0.49 and 0.54 or so
0.41 and 0.51 or so, it is speckless.
Method of the technical scheme 2. according to technical scheme 1, it is characterised in that：The step 1) described in methyl alcohol or second
Alcoholic solution refers to the solution being made after mixing methyl alcohol or ethanol with certain proportion with water, and e.g. 30%~100% methyl alcohol is molten
Liquid such as be 30%~100% ethanol solution, e.g. 60%~85% methanol solution such as be 60%~85%
Ethanol solution；
Method of the technical scheme 3. according to technical scheme 1, step 2) described in medicinal material coarse powder can be 20~100
The powder of mesh sieve；The meal is appropriate, can be 0.5~2g；
Method of the technical scheme 4. according to technical scheme 1, step 2) described in solvent refer to by methyl alcohol or ethanol with
Water mixed with certain proportion after the solution that is made, e.g. 30%~100% methanol solution such as be 30%~100% second
Alcoholic solution, e.g. 60%~85% methanol solution such as be 60%~85% ethanol solution；
Method of the technical scheme 5. according to technical scheme 1, step 3) described in take control medicinal material solution or test sample
Appropriate solution, can be 1~20 μ l, such as 5~15 μ l；The reversed phase thin layer plate, can be prefabricated reversed phase thin layer plate, and it is can
With what is obtained from commercially available approach；
Method of the technical scheme 6. according to technical scheme 1, step 4) described in 0.1% phosphate aqueous solution and ethanol with
Certain proportion is solvent, and the volume ratio of 0.1% phosphate aqueous solution and ethanol can be 3:7~7:3, such as 5：3~5, for example
5：4；
Method of the technical scheme 7. according to technical scheme 1, step 4) described in launch, can be One step development or two
Secondary expansion.
Method of the technical scheme 8. according to technical scheme 1, step 1) it is substantially according to following manner operation：
Radix Glycyrrhizae control medicinal material, Radix Glycyrrhizae control medicinal material (such as each 0.5~2g) are weighed, is respectively placed in conical flask with cover, plus
Solution (such as 30%~100% methanol solution, e.g. 60%~85% methanol solution, methyl alcohol account for volume ratio in the aqueous solution) or
(such as 30%~100% ethanol solution, e.g. 60%~85% ethanol solution, ethanol account for volume in the aqueous solution to ethanol solution
Than) appropriate (such as 50~200ml, such as 80~180ml), close plug, ultrasonically treated (such as power 500W, frequency 40kHz) example
Such as such as 30 minutes 20-40 minutes, let cool, filter, subsequent filtrate is taken, as control medicinal material solution.
Method of the technical scheme 9. according to technical scheme 1, step 2) medicinal material coarse powder is substantially according to such as lower section
What formula was prepared：Dry Radix Glycyrrhizae, swollen fruit Radix, glycyrrhiza glabra medicinal material are crushed respectively with high speed Universal pulverizer,
Cross 20~100 mesh sieves and obtain medicinal powder, be meal.
Method of the technical scheme 10. according to technical scheme 1, step 2) three kinds of need testing solutions are substantially photograph
What following manner was prepared：Radix Glycyrrhizae, swollen fruit Radix, glycyrrhiza glabra 0.5~2g of powder are weighed respectively, are respectively placed in tool plug taper
In bottle, plus 30%~100% (such as 60%~85%) methyl alcohol (methyl alcohol accounts for volume ratio in the aqueous solution) or 30%~100% (example
Such as 60%~85%) ethanol (ethanol accounts for volume ratio in the aqueous solution) it is appropriate (such as 50~200ml, such as 80~180ml), it is close
Plug, ultrasonically treated (power 500W, frequency 40kHz) such as 30 minutes such as 20-40 minutes 5~60 minutes lets cool, filtration, takes
Subsequent filtrate, is obtained Radix Glycyrrhizae, swollen fruit Radix, glycyrrhiza glabra need testing solution respectively.
Method of the technical scheme 11. according to technical scheme 1, step 4) it is described launch be in double flute Development of Thin-Layer Chromatography
Carried out in cylinder.
Method of the technical scheme 12. according to technical scheme 1, step 4) expansion substantially photograph following steps a)
Carried out with the mode of step b)：
A) 0.1% phosphate aqueous solution and ethanol are taken, according to volume ratio 5:4 are well mixed, and obtain solvent；Solvent is put
In first groove of the double flute thin layer chromatograph developing cylinder of dried and clean；
B) by step 3) reversed phase thin layer plate after point sample, it is placed in second groove of double flute thin layer chromatograph developing cylinder of step a)
In, closed cylinder cap, after presaturation 2~30 minutes (such as presaturation 10~15 minutes), by above-mentioned double flute thin layer chromatograph developing cylinder
Solvent in first groove is transferred in second groove for being placed with reversed phase thin layer plate, is launched, and exhibition terminates exhibition away from 10cm
Open, take out lamellae, dry, complete to launch.
Method of the technical scheme 13. according to technical scheme 1, it is as described in the embodiment of the present invention 1.
Method of the technical scheme 14. according to technical scheme 1, it is as described in the embodiment of the present invention 2.
Can be mutually combined to form new technical scheme between any two of the present invention or multiple technical schemes.
Any technical characteristic that any embodiment of either side of the present invention or the either side has is equally applicable
Any embodiment of other any embodiments or other either sides, as long as they will not be conflicting, certainly mutual
Between where applicable, if necessary can individual features be made with appropriate modification.Make into one with feature to various aspects of the present invention below
The description of step.
All documents recited in the present invention, their full content is incorporated herein by reference, and if these are literary
Offer expressed implication with it is of the invention inconsistent when, be defined by statement of the invention.Additionally, the various terms that use of the present invention and
Phrase has well known to a person skilled in the art general sense, nonetheless, the present invention remain desirable at this to these terms and
Phrase is described in more detail and explains, the term and phrase for referring to if any inconsistent with common art-recognized meanings, with institute's table of the present invention
The implication stated is defined.
Prior art discloses some to the various sides that Radix Glycyrrhizae, swollen fruit Radix, glycyrrhiza glabra three are made a distinction/differentiated
Method.For example, version in 2015《Chinese Pharmacopoeia》The Radix Glycyrrhizae in three kinds of sources is carried out from the angle of proterties in one " Radix Glycyrrhizae " recorded
Distinguish, but this differentiation requirement has the extremely rich experience could to complete, and common practitioner is to be difficult to carry out them
Scientifically distinguish.In addition, Cui Shufen (Cui Shufen etc., the research of micro emulsion thin-layered chromatography discriminating Radix Glycyrrhizae, Chinese herbal medicine, 2007
04 phase) a kind of thin-layered chromatography using micro emulsion solvent is disclosed to differentiate three kinds of Radix Glycyrrhizaes, wherein the micro emulsion composition for using is：
The normal heptane of+11% methyl alcohol of SDS+16% n-butanols+2.4% of 0.23mol/L, and need to use liquiritin and licochalcone
Two kinds of control monomeric compounds of first, the expansion plate that it is used also is polyamide film；Obviously, on the one hand the refined fragrant method of Cui exists
It is to want big more than the ethanol solution of the present invention micro phosphoric acid of addition to prepare the difficulty in terms of micro emulsion, for example in order that n-butanol,
Both normal heptanes are dissolved in the water, it is necessary to add surfactants/emulsifiers lauryl sodium sulfate i.e. SDS, and this micro-
Breast is unstable, there is the time limit for using；Further, since having used methyl alcohol, n-butanol, normal heptane these organic solvents, its is right
Cleaning etc. can all bring very big trouble after the health care of operator, environment-friendly, experiment；Furthermore, the method for Cui Shufen
Need to use two kinds of control monomeric compounds of liquiritin and Licochalcone A, and this control is originated with monomeric compound
Less, it is expensive, it is not that common laboratory can be equipped with.And the present invention uses micro phosphoric acid-ethanol-water system to enter as spread agent
The inverse thin layer chromatography of all very easy disposal of the aspect such as cleaning after row health care, environment-friendly, experiment, can be without right
It is to be capable of achieving three kinds of fast and effectively discriminatings of medicinal material to distinguish according to monomeric compound, this is that prior art all cannot be with the present invention
What technique effect was compared.Even in addition, the present inventor it has by the present invention been found that the inventive method, be also it is a kind of very
The unique and accurate method condition for selecting；For example, working as in Example 1 and Example 2 of the present invention only by solvent therein
Phosphoric acid used is changed to use the pH of the organic acids such as inorganic acid or formic acid or acetic acid such as hydrochloric acid, sulfuric acid or nitric acid and aqueous acid
When value is identical with the phosphate aqueous solution, the sapphire fluorescence spot at the place of Rf values 0.41 or so disappears, and only implements in the present invention
Example 1 and embodiment 2 are using phosphoric acid as could separate/show the place of Rf values 0.41 or so under conditions of solvent acidic components
Sapphire fluorescence spot；And for example, phosphate aqueous solution during only agent ought be spread out in Example 1 and Example 2 of the present invention
Volume ratio with ethanol is changed to 5：When changing in the range of 3~5, glycyrrhiza glabra is glimmering in two yellow greens of Rf values 0.49 and 0.54
Light spot can be separated, but if in only spreading out agent in Example 1 and Example 2 of the present invention phosphate aqueous solution with
The volume ratio of ethanol is changed to 5：Range such as aqueous outside 3~5 are than alcohol liquid volume ratio 5：2 or 5：When 6, light fruit is sweet
Two yellow-green fluorescence spots of the grass at Rf values 0.49 and 0.54 cannot efficiently separate and come and move on at Rf values about 0.51
A yellow-green fluorescence spot is presented, i.e., its in Fig. 1 the spot of B, D two move on at C, show i.e. using being chromatographic condition of the present invention
The trickle change of the species and water/proportion of ethanol of middle acid substance may have a strong impact on three kinds of discriminatings of Radix Glycyrrhizae and distinguish；Separately
Outward, it has been found that, when in Example 1 and Example 2 of the present invention, other analysis conditions present invention provide that in the range of it is appropriate
Three kinds of discriminatings of Radix Glycyrrhizae are not interfered with during change and distinguishes effect.For another example, wood closes diligent (the wood conjunction Bu Liabuli of Bu Liabuli
Diligent etc., Xinjiang Medicine University's journal, 02 phase in 2014) a kind of easy rapid identification method of swollen fruit Radix is disclosed, used by it
Method the total flavonoid extract of Xinjiang swollen fruit Radix and glycyrrhiza glabra is prepared using the ethanol extraction method of ultrasonic wave added；Utilize
Silica gel thin-layer chromatography analyzes 2 kinds of blob features of glycyrrhiza total flavonoid extract；Separated on lamellae using thin-layer method is prepared
Special spot simultaneously carries out structural analysis to separated monomer component, as a result shows that swollen fruit Radix ethanol extract is in solvent
Ethyl acetate: formic acid: shown a kind of special glassy yellow spot is accredited as Radix Glycyrrhizae under the conditions of the thin-layer chromatography of water (8: 3: 1)
Chalcone A, chromatographic characteristics are different from other Radix Glycyrrhizae kinds, show that the silica gel with licochalcone A as reference substance is fought layer chromatography
It is the short-cut method of quick discriminating swollen fruit Radix, is suitable for Rapid identification and the resource survey research of Radix Glycyrrhizae kind.But wood is closed
This literature method of cloth power is not appropriate for discriminating while three kinds of Radix Glycyrrhizaes, and due to which using substantial amounts of acetic acid second
Ester, formic acid, also use this monomer chemistries material of licochalcone A, and these method conditions can like that trigger order with Cui Shufen
People is difficult to the defect for receiving.
The advantage of the invention is that：The present invention differentiates that Radix Glycyrrhizae, swollen fruit Radix, light fruit are sweet using reverse phase thin layer chromatography first
Grass, method is simple to operate, and the instrument and equipment and control medicinal material popularization for being used are good, are easy to processing of crude drugs, sale enterprise and medicine
The popularization and application of product Inspection Unit.
Brief description of the drawings
Fig. 1 is the inverse thin layer chromatography discrimination method chromatogram (the ultraviolet light of 365nm) of the resulting tape of embodiment 1 mark, figure
In 1 be Radix Glycyrrhizae (swollen fruit Radix) control medicinal material, 2 be swollen fruit Radix test sample, 3 be glycyrrhiza glabra test sample, 4 be Radix Glycyrrhizae (Radix Glycyrrhizae)
Control medicinal material, 5 is Radix Glycyrrhizae test sample；Specifically, being shown in figure 1 above：
Radix Glycyrrhizae test sample and Radix Glycyrrhizae control medicinal material in Rf values 0.41 or so (the corresponding positions of A in figure), have one it is sky blue
The fluorescence spot of color, in Rf values 0.51 or so (the corresponding positions of C in figure), there is a fluorescence spot for yellow green；
Swollen fruit Radix and swollen fruit Radix control medicinal material only in Rf values 0.41 or so (the corresponding positions of A in figure), there is one
Sapphire fluorescence spot；
Glycyrrhiza glabra respectively has a fluorescence for yellow green in Rf values 0.49 and 0.54 or so (the corresponding positions of B and D in figure)
Spot, in Rf values 0.41 and 0.51 or so (the corresponding positions of A and C in figure), is speckless.
Fig. 2 is the gained of embodiment 2 (not with mark) inverse thin layer chromatography discrimination method chromatogram (365nm uviol lamps
Light), 1~5 material for representing is identical with the material that in Fig. 1 1~5 represents in figure.
Specific embodiment
The present invention can be conducted further description by the following examples, however, the scope of the present invention is not limited
In following embodiments.One of skill in the art, can be with it is understood that on the premise of without departing substantially from the spirit and scope of the present invention
Various change and modification are carried out to the present invention.The present invention to used in experiment to material and test method carry out generality
And/or specific description.Although for realize many materials that the object of the invention used and operating method be it is known in the art that
But the present invention is still described in detail as far as possible herein.Following examples further illustrate the present invention, rather than limiting this hair
It is bright.
Embodiment 1：Reverse phase thin layer chromatography differentiates Radix Glycyrrhizae, swollen fruit Radix, the method for glycyrrhiza glabra.
1) preparation of control medicinal material solution：
Radix Glycyrrhizae (Radix Glycyrrhizae) control medicinal material, Radix Glycyrrhizae (swollen fruit Radix) each 1g of control medicinal material are weighed respectively, are respectively placed in tool plug cone
In shape bottle, plus 70% ethanol (ethanol accounts for volume ratio in the aqueous solution) 100ml, close plug, ultrasonically treated (power 500W, frequency
40kHz) 30 minutes, let cool, filter, take subsequent filtrate, it is standby as control medicinal material solution；
2) prepared by medicinal material need testing solution：
Dry Radix Glycyrrhizae, swollen fruit Radix, glycyrrhiza glabra medicinal material are crushed respectively with high speed Universal pulverizer, crosses 60 mesh
Sieve to obtain medicinal powder；
Radix Glycyrrhizae, swollen fruit Radix, glycyrrhiza glabra powder 1g are weighed, is respectively placed in conical flask with cover, plus 70% ethanol (ethanol
Account for volume ratio in the aqueous solution) 100ml, close plug, ultrasonically treated (power 500W, frequency 40kHz) 30 minutes lets cool, filtration, takes continuous
Filtrate, is obtained Radix Glycyrrhizae, swollen fruit Radix, glycyrrhiza glabra need testing solution.
3) 2 kinds of control medicinal material solution obtained above and 3 kinds of each 10 μ l of need testing solution are drawn respectively with quantitative capillary tube,
Same horizontal line is not in same reversed phase thin layer plate (this inverse thin layer chromatography plate directly can be bought from market) for point sample successively
With on position.
4) method of deploying, is operated by following a~b methods：
A) 0.1% phosphate aqueous solution and ethanol are taken, according to volume ratio 5:4 are well mixed, and obtain solvent.Solvent is put
In first groove of the double flute thin layer chromatograph developing cylinder of dried and clean；
B) by step 3) reversed phase thin layer plate after point sample, it is placed in second groove of double flute thin layer chromatograph developing cylinder of step a)
In, closed cylinder cap, after presaturation 10~15 minutes, by the solvent transfer in first groove of above-mentioned double flute thin layer chromatograph developing cylinder
To being placed with second groove of reversed phase thin layer plate, launched, exhibition terminates to launch away from 10cm, takes out lamellae, dries, and completes
Launch；
5) spot in chromatogram is compared as follows, so as to differentiate Radix Glycyrrhizae, swollen fruit Radix, glycyrrhiza glabra：
In reviewing step 4 under 365nm length ultraviolet lamps) reversed phase thin layer plate b), and compare the fluorescence spot in chromatogram
(referring to accompanying drawing), to differentiate Radix Glycyrrhizae, swollen fruit Radix, glycyrrhiza glabra；
Radix Glycyrrhizae test sample and Radix Glycyrrhizae (Radix Glycyrrhizae) control medicinal material have one in Rf values 0.41 or so (the corresponding positions of A in figure)
Individual sapphire fluorescence spot, in Rf values 0.51 or so (the corresponding positions of C in figure), there is a fluorescence spot for yellow green；
Swollen fruit Radix and Radix Glycyrrhizae (swollen fruit Radix) control medicinal material only in Rf values 0.41 or so (the corresponding positions of A in figure),
There is a sapphire fluorescence spot；
Glycyrrhiza glabra respectively has a fluorescence for yellow green in Rf values 0.49 and 0.54 or so (the corresponding positions of B and D in figure)
Spot, in Rf values 0.41 and 0.51 or so (the corresponding positions of A and C in figure), is speckless.
Fig. 1 shows that the present embodiment inspects figure under 365nm wavelength.In addition, the gained thin layer collection of illustrative plates of Examples below 2 is
Fig. 2 is essentially identical with the gained Fig. 1 of above-described embodiment 1.
Embodiment 2：Reverse phase thin layer chromatography differentiates Radix Glycyrrhizae, swollen fruit Radix, the method for glycyrrhiza glabra.
1) preparation of control medicinal material solution：
Radix Glycyrrhizae (Radix Glycyrrhizae) control medicinal material, Radix Glycyrrhizae (swollen fruit Radix) each 2g of control medicinal material are weighed respectively, are respectively placed in tool plug cone
In shape bottle, plus 80% methyl alcohol (methyl alcohol accounts for volume ratio in the aqueous solution) 150ml, close plug, ultrasonically treated (power 500W, frequency
40kHz) 30 minutes, let cool, filter, take subsequent filtrate, it is standby as control medicinal material solution；
2) prepared by medicinal material need testing solution：
Dry Radix Glycyrrhizae, swollen fruit Radix, glycyrrhiza glabra medicinal material are crushed respectively with high speed Universal pulverizer, crosses 60 mesh
Sieve to obtain medicinal powder；
Radix Glycyrrhizae, swollen fruit Radix, glycyrrhiza glabra powder 2g are weighed respectively, are respectively placed in conical flask with cover, plus 80% methyl alcohol
(methyl alcohol accounts for volume ratio in the aqueous solution) 150ml, close plug, ultrasonically treated (power 500W, frequency 40kHz) 30 minutes lets cool, filter
Cross, take subsequent filtrate, Radix Glycyrrhizae, swollen fruit Radix, glycyrrhiza glabra need testing solution is obtained.
3) 2 kinds of control medicinal material solution obtained above and 3 kinds of each 10 μ l of need testing solution are drawn respectively with quantitative capillary tube,
Point sample is on the diverse location of same reversed phase thin layer plate same horizontal line successively.
4) method of deploying, is operated by following a~b methods：
A) 0.1% phosphate aqueous solution and ethanol are taken, according to volume ratio 5:4 are well mixed, and obtain solvent.Solvent is put
In first groove of the double flute thin layer chromatograph developing cylinder of dried and clean；
B) by step 3) reversed phase thin layer plate after point sample, it is placed in second groove of double flute thin layer chromatograph developing cylinder of step a)
In, closed cylinder cap, after presaturation 10~15 minutes, by the solvent transfer in first groove of above-mentioned double flute thin layer chromatograph developing cylinder
To being placed with second groove of reversed phase thin layer plate, launched, exhibition terminates to launch away from 10cm, takes out lamellae, dries, and completes
Launch；
5) spot compared as follows in chromatogram differentiates Radix Glycyrrhizae, swollen fruit Radix, glycyrrhiza glabra：
In reviewing step 4 under 365nm length ultraviolet lamps) reversed phase thin layer plate b), and compare the fluorescence spot in chromatogram
(referring to accompanying drawing), to differentiate Radix Glycyrrhizae, swollen fruit Radix, glycyrrhiza glabra；
Radix Glycyrrhizae test sample and Radix Glycyrrhizae (Radix Glycyrrhizae) control medicinal material have one in Rf values 0.41 or so (the corresponding positions of A in figure)
Individual sapphire fluorescence spot, in Rf values 0.51 or so (the corresponding positions of C in figure), there is a fluorescence spot for yellow green；
Swollen fruit Radix and Radix Glycyrrhizae (swollen fruit Radix) control medicinal material only in Rf values 0.41 or so (the corresponding positions of A in figure),
There is a sapphire fluorescence spot；
Glycyrrhiza glabra respectively has a fluorescence for yellow green in Rf values 0.49 and 0.54 or so (the corresponding positions of B and D in figure)
Spot, in Rf values 0.41 and 0.51 or so (the corresponding positions of A and C in figure), is speckless.
Embodiment described above is only the preferred embodiment lifted to absolutely prove the present invention, protection model of the invention
Enclose not limited to this.Equivalent substitute or conversion that those skilled in the art are made on the basis of the present invention, in the present invention
Protection domain within.Protection scope of the present invention is defined by claims.
Claims (10)
1. reverse phase thin layer chromatography differentiates Radix Glycyrrhizae, swollen fruit Radix, the method for glycyrrhiza glabra, it is characterised in that comprise the following steps：
1) Radix Glycyrrhizae control medicinal material, swollen fruit Radix control medicinal material are weighed respectively, plus methyl alcohol or ethanol solution are made control medicinal material solution；
2) extracting liquorice, swollen fruit Radix, three kinds of medicinal material coarse powders of glycyrrhiza glabra are appropriate, are respectively placed in conical flask with cover, solubilizer, surpass
Sonication (such as 5~60 minutes), filtration, take subsequent filtrate, respectively be obtained Radix Glycyrrhizae, swollen fruit Radix, glycyrrhiza glabra test sample it is molten
Liquid；
3) with quantitative capillary tube distinguish aspiration step 1) obtained in two kinds of control medicinal material solution and step 2) obtained in three kinds for examination
Product solution is appropriate, and point sample is on the same horizontal line diverse location of same reversed phase thin layer plate successively；
4) with 0.1% phosphate aqueous solution and ethanol with certain proportion as solvent, launched, taken out after expansion, dried；
5) spot compared as follows in chromatogram differentiates Radix Glycyrrhizae, swollen fruit Radix, glycyrrhiza glabra：
In reviewing step 4 under 365nm length ultraviolet lamps) gained reversed phase thin layer plate, and compare the fluorescence spot in chromatogram, to reflect
Other Radix Glycyrrhizae, swollen fruit Radix, glycyrrhiza glabra；
Radix Glycyrrhizae test sample and Radix Glycyrrhizae control medicinal material have a sapphire fluorescence spot at the place of Rf value Rf values 0.41 or so,
At the place of Rf values 0.51 or so, there is a fluorescence spot for yellow green；
Swollen fruit Radix test sample and swollen fruit Radix control medicinal material only at the place of Rf values 0.41 or so, there is a sapphire fluorescent spot
Point；
Glycyrrhiza glabra test sample respectively has a fluorescence spot for yellow green, in Rf values 0.41 at the place of Rf values 0.49 and 0.54 or so
With 0.51 or so, it is speckless.
2. method according to claim 1, it is characterised in that：The step 1) described in methyl alcohol or ethanol solution refer to by
The solution that methyl alcohol or ethanol are made after being mixed with certain proportion with water, e.g. 30%~100% methanol solution such as be
30%~100% ethanol solution, e.g. 60%~85% methanol solution such as be 60%~85% ethanol solution.
3. method according to claim 1, step 2) described in medicinal material coarse powder can be 20~100 mesh sieves powder；
The meal is appropriate, can be 0.5~2g.
4. method according to claim 1, step 2) described in solvent refer to certain proportion by methyl alcohol or ethanol with water
The solution being made after mixing, e.g. 30%~100% methanol solution such as be 30%~100% ethanol solution, for example
It is 60%~85% methanol solution such as is 60%~85% ethanol solution.
5. method according to claim 1, step 3) described in take control medicinal material solution or appropriate need testing solution, can be with
It is 1~20 μ l, such as 5~15 μ l；The reversed phase thin layer plate, can be prefabricated reversed phase thin layer plate, and it can be from commercially available approach
Obtain.
6. method according to claim 1, step 4) described in 0.1% phosphate aqueous solution and ethanol with certain proportion be exhibition
Open agent, the volume ratio of 0.1% phosphate aqueous solution and ethanol can be 3:7~7:3, such as 5：3~5, such as 5：4.
7. method according to claim 1, step 4) described in launch, can be One step development or second outspread.
8. method according to claim 1, step 1) it is substantially according to following manner operation：
Radix Glycyrrhizae control medicinal material, Radix Glycyrrhizae control medicinal material (such as each 0.5~2g) are weighed, is respectively placed in conical flask with cover, solubilization liquid
(such as 30%~100% methanol solution, e.g. 60%~85% methanol solution, methyl alcohol account for volume ratio in the aqueous solution) or ethanol
Solution (such as 30%~100% ethanol solution, e.g. 60%~85% ethanol solution, ethanol account for volume ratio in the aqueous solution) is fitted
Amount (such as 50~200ml, such as 80~180ml), close plug, ultrasonically treated (such as power 500W, frequency 40kHz) such as 20-
Such as 30 minutes 40 minutes, let cool, filter, subsequent filtrate is taken, as control medicinal material solution.
9. method according to claim 1, step 2) medicinal material coarse powder is substantially to be prepared according to following manner：
Dry Radix Glycyrrhizae, swollen fruit Radix, glycyrrhiza glabra medicinal material are crushed respectively with high speed Universal pulverizer, crosses 20~100 mesh sieves
Medicinal powder is obtained, is meal.
10. method according to claim 1, it is characterised in that：
Step 2) three kinds of need testing solutions are substantially to be prepared according to following manner：It is sweet that Radix Glycyrrhizae, swollen fruit are weighed respectively
Grass, glycyrrhiza glabra 0.5~2g of powder, are respectively placed in conical flask with cover, plus 30%~100% (such as 60%~85%) methyl alcohol
(methyl alcohol accounts for volume ratio in the aqueous solution) or 30%~100% (such as 60%~85%) ethanol (ethanol accounts for volume ratio in the aqueous solution)
In right amount (such as 50~200ml, such as 80~180ml), close plug, ultrasonically treated (power 500W, frequency 40kHz) 5~60 minutes
Such as 30 minutes such as 20-40 minutes, let cool, filter, take subsequent filtrate, Radix Glycyrrhizae, swollen fruit Radix, glycyrrhiza glabra are obtained respectively for examination
Product solution；
Step 4) it is described expansion carried out in double flute thin layer chromatograph developing cylinder；
Step 4) described launch what is substantially carried out according to the mode of following steps a) and step b)：
A) 0.1% phosphate aqueous solution and ethanol are taken, according to volume ratio 5:4 are well mixed, and obtain solvent；Solvent is put into drying
In first groove of clean double flute thin layer chromatograph developing cylinder；
B) by step 3) reversed phase thin layer plate after point sample, it is placed in second groove of double flute thin layer chromatograph developing cylinder of step a), it is close
Cylinder cap is closed, after presaturation 2~30 minutes (such as presaturation 10~15 minutes), by above-mentioned double flute thin layer chromatograph developing cylinder first
Solvent in groove is transferred in second groove for being placed with reversed phase thin layer plate, is launched, and exhibition terminates to launch away from 10cm, takes out
Lamellae, dries, and completes to launch；
It is as described in the embodiment of the present invention 1；And/or
It is as described in the embodiment of the present invention 2.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100729437B1 (en) * | 2006-02-22 | 2007-06-27 | 한국생명공학연구원 | The extracts of corylopsis coreana and tellimagrandin i isolated from same having antifungal activity |
| CN102078386A (en) * | 2009-11-30 | 2011-06-01 | 江西济民可信药业有限公司 | Method for detecting Xiaoyao mixture |
| US20110239745A1 (en) * | 2010-04-01 | 2011-10-06 | Lawrence Livermore National Security, Llc | Rapid Identification of Explosives Using Thin-Layer Chromatography and Colorimetric Techniques |
| CN105063041A (en) * | 2015-09-10 | 2015-11-18 | 山西农业大学 | SSR molecular marker primer for detecting glycyrrhiza plants |
| CN105136967A (en) * | 2015-07-23 | 2015-12-09 | 北京市药品检验所 | Reversed-phase thin-layer chromatographic method for identifying florists chrysanthemum and bo chrysanthemum |
-
2017
- 2017-03-08 CN CN201710134869.9A patent/CN106841501B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100729437B1 (en) * | 2006-02-22 | 2007-06-27 | 한국생명공학연구원 | The extracts of corylopsis coreana and tellimagrandin i isolated from same having antifungal activity |
| CN102078386A (en) * | 2009-11-30 | 2011-06-01 | 江西济民可信药业有限公司 | Method for detecting Xiaoyao mixture |
| US20110239745A1 (en) * | 2010-04-01 | 2011-10-06 | Lawrence Livermore National Security, Llc | Rapid Identification of Explosives Using Thin-Layer Chromatography and Colorimetric Techniques |
| CN105136967A (en) * | 2015-07-23 | 2015-12-09 | 北京市药品检验所 | Reversed-phase thin-layer chromatographic method for identifying florists chrysanthemum and bo chrysanthemum |
| CN105063041A (en) * | 2015-09-10 | 2015-11-18 | 山西农业大学 | SSR molecular marker primer for detecting glycyrrhiza plants |
Non-Patent Citations (4)
| Title |
|---|
| SUENGMOK CHO 等: "Hypnotic effects and GABAergic mechanism of licorice (Glycyrrhiza glabra) ethanol extract and its major flavonoid constituent glabrol", 《BIOORGANIC & MEDICINAL CHEMISTRY》 * |
| 冯雅斌 等: "薄层色谱法在药物分析中的应用及研究进展", 《疾病监测与控制杂志》 * |
| 崔淑芬 等: "微乳薄层色谱法鉴别甘草的研究", 《中草药》 * |
| 木合布力·阿布力孜 等: "胀果甘草的简便快速鉴定方法", 《新疆医科大学学报》 * |
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