CN108982741A - The discrimination method of method semen coicis based on thin-layer chromatography - Google Patents
The discrimination method of method semen coicis based on thin-layer chromatography Download PDFInfo
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Abstract
The present invention provides a kind of discrimination method of method semen coicis based on thin-layer chromatography, and the discrimination method includes: to prepare test solution；Follow the example of semen coicis control medicinal material；Take the test solution, reference substance solution, it is put respectively on same lamellae, using volume ratio for (75~82): (15~20): 1 petroleum ether, ether, glacial acetic acid mixture be unfolded as solvent, lamellae, spray color developing agent, heating colour developing are dried, is inspected under ultraviolet lamp.The present invention is based on study for a long period of time to method semen coicis, when being identified using thin-layered chromatography to it, selecting volume ratio is (75~82): (15~20): 1 petroleum ether, ether, glacial acetic acid mixture be that solvent is unfolded, using the solvent of the special formulation, it is good that effect is unfolded, without repeating expansion operation or carrying out presaturation to expansion cylinder, simplifies and identify distance, identification efficiency is high.
Description
Technical field
The present invention relates to traditional Chinese medicine quality detection technique fields, more particularly to a kind of method semen coicis based on thin-layer chromatography
Discrimination method.
Background technique
Thin-layered chromatography (TLC) is a kind of micro, quick, easy method for separating and analyzing.System applies suitable stationary phase
It is distributed on glass plate, plastics or aluminium substrate, at a thin uniform layer.After point sample, expansion, according to Rf value (Rf) and suitable pair
It is compared according to the Rf value of object resulting chromatogram by the same method, to carry out the identification of drug, determination of foreign matter or assay
Method.This method is a kind of critically important experimental technique of quick separating and a small amount of substance of qualitative analysis, is also used for tracking reaction
Process.Have the characteristics that duration of run is short, separating capacity is strong, high sensitivity.
Semen coicis is gramineae Coix lacryma-jobi L.var.mayuen (Roman.) Stapf's
Dry mature kernal.As Chinese medicine, semen coicis have wet sharp water penetration, invigorating the spleen to arrest diarrhea, eliminating impediment, apocenosis, detoxicating and resolving a mass work
With.The traditional concocting method of semen coicis be frying, generally fry, bran fry, soil fry etc., after processing gained semen coicis product, example
Such as method semen coicis, the content of glycerol trioleate therein can be risen, are conducive to the promotion of drug effect, other chemical components
Type and content can may also accordingly become complicated.And the variation of chemical composition group, the band of semen coicis product is stranded
It disturbs.When traditional use thin-layered chromatography identifies method semen coicis, it may appear that expansion effect is poor, such as spot is fuzzy, spot
Point hangover is serious, even detects the defect of not spotting out.In order to obtain good expansion effect, in thin layer chromatography analysis,
Presaturation processing would generally be carried out in advance to expansion cylinder using repeatedly expansion, or using solvent.But above-mentioned processing method
Gained thin-layer chromatogram effect be not still it is especially good, and cumbersome, identification efficiency is low.
Summary of the invention
Based on this, the main object of the present invention is to provide that a kind of expansion effect is good, identification efficiency is high based on thin-layer chromatography
Method semen coicis discrimination method.
The purpose of the present invention is what is be achieved through the following technical solutions:
A kind of discrimination method of the method semen coicis based on thin-layer chromatography, the discrimination method include:
Method semen coicis to be measured is taken, test solution is prepared；
Semen coicis control medicinal material is followed the example of, reference substance solution is prepared；
Thin-layer chromatography: taking the test solution, reference substance solution, is put respectively on same lamellae, with volume ratio
For (75~82): (15~20): 1 petroleum ether, ether, glacial acetic acid mixture be that solvent is unfolded, dry lamellae,
Color developing agent, heating colour developing are sprayed, is inspected under ultraviolet lamp.
It is (80~82): (15~18) that the solvent, which is volume ratio, in one of the embodiments: 1 petroleum ether,
The mixture of ether, glacial acetic acid.
In one of the embodiments, the solvent be volume ratio be 82:18:1 petroleum ether, ether, glacial acetic acid
Mixture.
The lamellae uses sodium carboxymethylcellulose for the silica G thin layer of binder in one of the embodiments,
Plate；The color developing agent is the mixture of sulfuric acid and ethyl alcohol, and the volume of sulfuric acid accounts for 8%~12% in the mixture.
The color developing agent is the mixture of sulfuric acid and ethyl alcohol, sulfuric acid in the mixture in one of the embodiments,
Volume account for 10%.
Test solution is prepared described in one of the embodiments, is included the following steps:
Method semen coicis to be measured is taken, powder is made, the petroleum ether that temperature is 60 DEG C~90 DEG C is added, is ultrasonically treated, filtering, institute
Obtaining filtrate is test solution；
The preparation reference substance solution, includes the following steps:
Semen coicis control medicinal material is followed the example of, powder is made, the petroleum ether that temperature is 60 DEG C~90 DEG C is added, ultrasonic treatment, mistake
Filter, gained filtrate is reference substance solution.
The temperature of the heating colour developing is 103~106 DEG C in one of the embodiments,.
The ultraviolet light wavelength is 365nm in one of the embodiments,.
The method semen coicis includes the method semen coicis medicine materical crude slice for the method that is prepared by the following procedure in one of the embodiments:
(1) clean semen coicis, boiling are taken；
(2) semen coicis obtained by step (1) is pulled out, draining is added edible oil, mixes thoroughly；
(3) semen coicis obtained by step (2) is steamed to the well-done heart, taking-up is dried, light shines；
(4) semen coicis obtained by sand stir-fry step (3) is flower-shaped to rice krispies, shakeout.
In one of the embodiments, in step (1), the time of the boiling is 10~20min；It is described in step (2)
Edible oil is at least one of peanut oil, tea-seed oil；Semen coicis obtained by every 100kg step (1) be added the edible oil 2.5~
2.8kg；Or/and the edible oil is selected from least one of peanut oil, tea-seed oil, castor oil.
Compared with prior art, the present invention have it is following the utility model has the advantages that
The present invention is based on studying for a long period of time to method semen coicis, when identifying using thin-layered chromatography to it, body is selected
Product is than being (75~82): (15~20): 1 petroleum ether, ether, glacial acetic acid mixture be that solvent is unfolded, using this
The solvent of special formulation, expansion effect is good, without repeating expansion operation or carrying out presaturation to expansion cylinder, simplifies mirror
Other distance, identification efficiency are high.
Under conditions of the present invention is using the solvent of special formulation, cooperation uses sodium carboxymethylcellulose pyce for the silica gel of binder
G lamellae and color developing agent can shorten the time of expansion, colour developing.
Detailed description of the invention
Fig. 1 is the thin-layer chromatogram of embodiment 1；
Fig. 2 is the thin-layer chromatogram of embodiment 2；
Fig. 3 is the thin-layer chromatogram of embodiment 3.
Specific embodiment
It to facilitate the understanding of the present invention, below will be to invention is more fully described.But the present invention can be to be permitted
Mostly different form is realized, however it is not limited to embodiment described herein.On the contrary, purpose of providing these embodiments is makes
It is more thorough and comprehensive to the understanding of the disclosure.
Unless otherwise defined, all technical and scientific terms used herein and belong to technical field of the invention
The normally understood meaning of technical staff is identical.Term as used herein in the specification of the present invention is intended merely to description tool
The purpose of the embodiment of body, it is not intended that in the limitation present invention.Term as used herein "and/or" includes one or more phases
Any and all combinations of the listed item of pass.
Method semen coicis of the invention includes the semen coicis legal system medicine materical crude slice of following method preparation: (1) taking clean semen coicis, water
It boils；(2) moisture on semen coicis surface obtained by step (1) is drained, edible oil is added, mixes thoroughly；(3) by semen coicis obtained by step (2)
It steams to the well-done heart, taking-up is dried, light shines；(4) semen coicis obtained by sand stir-fry step (3) is flower-shaped to rice krispies, shakeout.
Specifically, semen coicis legal system medicine materical crude slice involved by following embodiments, including sample Y1 to sample Y9；Wherein:
Method acquisition is prepared by the following procedure in described sample Y1, Y4, the Y7: (1) taking semen coicis, cross 20 mesh~50 meshes
Hole removes granule and fragment etc., obtains clean semen coicis, and every kilogram of semen coicis adds water 3L to carry out boiling；The time of the boiling
For 10min；(2) moisture on semen coicis surface obtained by step (1) is drained, edible oil is added, mixes thoroughly；The edible oil is peanut
Oil；The edible oil 2.5kg is added in semen coicis obtained by every 100kg step (1)；(3) semen coicis obtained by step (2) is steamed to well-done
The heart, taking-up is dried, light shines；Described dry is that the semen coicis taking-up steamed to the well-done heart is placed at aeration-drying to dry in the air to surface
Existing gloss, color are bright；The light solarization is to be placed in the semen coicis after drying to shine under the sun to holding to have hardness；(4) sand is fried
Semen coicis obtained by step (3) is flower-shaped to rice krispies, shakeout；The condition that the sand is fried include: temperature be 140 DEG C, the time is
5min。
Method acquisition: the basic phase of preparation method with sample Y1 is prepared by the following procedure in described sample Y2, Y5, the Y8
Together, variation place is only that, in step (1), the time of boiling is 15min.
Method acquisition: the basic phase of preparation method with sample Y1 is prepared by the following procedure in described sample Y3, Y6, the Y9
Together, variation place is only that, in step (1), the time of boiling is 20min.
The semen coicis legal system medicine materical crude slice being prepared through this processing procedure, contained chemical composition group changes, therein
The content of glycerol trioleate can at least reach 1.5%.
Embodiment 1
The discrimination method of the present embodiment provides a kind of semen coicis legal system medicine materical crude slice based on thin-layer chromatography.
The semen coicis legal system medicine materical crude slice of the present embodiment, sample Y1 as described above, sample Y2, sample Y3.
The discrimination method of the present embodiment includes:
(1) prepared by test solution: taking semen coicis legal system medicine materical crude slice, powder is made, take 1g powder, add petroleum ether (60~90
DEG C) 10ml, it is ultrasonically treated 30 minutes, filtration takes filtrate, as test solution.
Accordingly prepare the test solution of the test solution of sample Y1, the test solution of sample Y2, sample Y3.
(2) reference substance solution prepare: separately take semen coicis control medicinal material (National Institute for Food and Drugs Control, lot number:
121254-201504), powder is made, takes 1g powder, adds petroleum ether (60~90 DEG C) 10ml, is ultrasonically treated 30 minutes, filtration,
Filtrate is taken, as reference substance solution.
(3) according to thin-layered chromatography test (2015 editions four general rules 0502 of Chinese Pharmacopoeia):
1) above-mentioned test solution, each 2 μ l of reference substance solution are drawn, being put respectively in same sodium carboxymethylcellulose is bonding
On the silica gel g thin-layer plate of agent；
2) with petroleum ether (60~90 DEG C)-ether-glacial acetic acid (82:18:1) for solvent, it is unfolded, takes out, dry；
3) it sprays with 10% ethanol solution of sulfuric acid, it is clear to be heated to spot development at 105 DEG C, inspects under 365nm ultraviolet lamp.
The present embodiment identification result is shown in Fig. 1, in Fig. 1,1,2,3 it is corresponding be sample Y1, sample Y2, sample Y3 it is thin
Layer chromatography figure, 10 be semen coicis control medicinal material thin-layer chromatogram, be can be seen that according to Fig. 1, and spot expansion effect is especially good, is not dragged
Tail clearly can perform well in the identification of method semen coicis.Also, step needed for the present embodiment obtains such expansion effect
It is rapid 2) in duration of run be 10min, heating developing time needed for step 3) is 120s.
Embodiment 2
The discrimination method of the present embodiment provides a kind of semen coicis legal system medicine materical crude slice based on thin-layer chromatography.
The semen coicis legal system medicine materical crude slice of the present embodiment, sample Y4 as described above, sample Y5, sample Y6.
The discrimination method of the present embodiment includes:
(1) prepared by test solution: with embodiment 1；
(2) prepared by reference substance solution: with embodiment 1；
(3) according to thin-layered chromatography test (2015 editions four general rules 0502 of Chinese Pharmacopoeia):
1) above-mentioned test solution, each 2 μ l of reference substance solution are drawn, being put respectively in same sodium carboxymethylcellulose is bonding
On the silica gel g thin-layer plate of agent；
2) with petroleum ether (60~90 DEG C)-ether-glacial acetic acid (80:15:1) for solvent, it is unfolded, takes out, dry；
3) it sprays with 8% ethanol solution of sulfuric acid, it is clear to be heated to spot development at 105 DEG C, inspects under 365nm ultraviolet lamp.
The present embodiment identification result is shown in Fig. 2, in Fig. 2,4,5,6 it is corresponding be sample Y4, sample Y5, sample Y6 it is thin
Layer chromatography figure, 10 be semen coicis control medicinal material thin-layer chromatogram, be can be seen that according to fig. 2, and spot expansion effect is preferable, is not dragged
Tail, clearly.Also, the duration of run in step 2) needed for the present embodiment obtains such expansion effect is 12min, step 3)
Required heating developing time is 150s.
Embodiment 3
The discrimination method of the present embodiment provides a kind of semen coicis legal system medicine materical crude slice based on thin-layer chromatography.
The semen coicis legal system medicine materical crude slice of the present embodiment, sample Y7 as described above, sample Y8, sample Y9.
The discrimination method of the present embodiment includes:
(1) prepared by test solution: with embodiment 1；
(2) prepared by reference substance solution: with embodiment 1；
(3) according to thin-layered chromatography test (2015 editions four general rules 0502 of Chinese Pharmacopoeia):
1) above-mentioned test solution, each 2 μ l of reference substance solution are drawn, being put respectively in same sodium carboxymethylcellulose is bonding
On the silica gel g thin-layer plate of agent；
2) with petroleum ether (60~90 DEG C)-ether-glacial acetic acid (75:20:1) for solvent, it is unfolded, takes out, dry；
3) it sprays with 12% ethanol solution of sulfuric acid, it is clear to be heated to spot development at 105 DEG C, inspects under 365nm ultraviolet lamp.
The present embodiment identification result is shown in Fig. 3, in Fig. 3,7,8,9 it is corresponding be sample Y7, sample Y8, sample Y9 it is thin
Layer chromatography figure, 10 be semen coicis control medicinal material thin-layer chromatogram, be can be seen that according to Fig. 3, and spot is unfolded, and does not trail, clearly.
Also, the duration of run in step 2) needed for the present embodiment obtains such expansion effect is 15min, is added needed for step 3)
Hot developing time is 160s.
Embodiment 4
The present embodiment is the change case of embodiment 3, provides a kind of identification of semen coicis legal system medicine materical crude slice based on thin-layer chromatography
Method.
The semen coicis legal system medicine materical crude slice of the present embodiment, sample Y7 as described above, sample Y8, sample Y9.
The discrimination method of the present embodiment includes:
(1) prepared by test solution: with embodiment 3；
(2) prepared by reference substance solution: with embodiment 3；
(3) according to thin-layered chromatography test (2015 editions four general rules 0502 of Chinese Pharmacopoeia):
1) above-mentioned test solution, each 2 μ l of reference substance solution are drawn, being put respectively in same sodium carboxymethylcellulose is bonding
On the silica gel g thin-layer plate of agent；
2) with petroleum ether (60~90 DEG C)-ether-glacial acetic acid (82:15:1) for solvent, it is unfolded, takes out, dry；
3) it sprays with 8% ethanol solution of sulfuric acid, it is clear to be heated to spot development at 105 DEG C, inspects under 365nm ultraviolet lamp.
The present embodiment identification result is the same as embodiment 3.
Comparative example 1
This comparative example is the comparative example of embodiment 1, and compared with Example 1, essential difference includes the solvent that uses for 10:
The petroleum ether of 3:0.1, ether, glacial acetic acid mixture, and the number for repeating that operation is unfolded is 3 times.
This comparative example the result is that: although expansion is repeatedly, the spot for characterizing method semen coicis chemical component to be measured does not have still
There is expansion, the thin-layer chromatography for discrimination can not be formed.
Comparative example 2
This comparative example is the comparative example of embodiment 1, and compared with Example 1, essential difference includes: in step 1) using without glutinous
The silica gel g thin-layer plate of mixture；Step 3) uses 20% ethanol solution of sulfuric acid
This comparative example the result is that: spot be unfolded effect it is especially good, do not trail, clearly, the method heart of a lotus seed can be performed well in
The identification of the seed of jog's tears.Also, the duration of run in step 2) needed for this comparative example obtains such expansion effect is 18min, step
It is rapid 3) needed for heating developing time be 180s.
According to the result description of above each example it is found that the ingredient of the good partition method semen coicis of the energy of embodiment 1 to 4, shape
At clearly spot.Identify in practice long-term, the separating effect that embodiment 1 is formed is best, and embodiment 2 is preferable, embodiment 3,4
Slightly take second place.This explanation, there are preferred embodiments for the formula of solvent.In addition, on this basis if the chromatoplate of appropriate mix and
Color developing agent, additionally it is possible to accelerate the time of expansion, colour developing.
In conclusion identifying using thin-layered chromatography to it the present invention is based on studying for a long period of time to method semen coicis
When, selecting volume ratio is (75~82): (15~20): 1 petroleum ether, ether, glacial acetic acid mixture be that solvent is opened up
It opens, using the solvent of the special formulation, expansion effect is good, without repeating expansion operation or carrying out to expansion cylinder pre- full
With, simplify and identifies distance, identification efficiency height.
Each technical characteristic of embodiment described above can be combined arbitrarily, for simplicity of description, not to above-mentioned reality
It applies all possible combination of each technical characteristic in example to be all described, as long as however, the combination of these technical characteristics is not deposited
In contradiction, all should be considered as described in this specification.
The embodiments described above only express several embodiments of the present invention, and the description thereof is more specific and detailed, but simultaneously
It cannot therefore be construed as limiting the scope of the patent.It should be pointed out that coming for those of ordinary skill in the art
It says, without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to protection of the invention
Range.Therefore, the scope of protection of the patent of the invention shall be subject to the appended claims.
Claims (10)
1. a kind of discrimination method of the method semen coicis based on thin-layer chromatography, which is characterized in that the discrimination method includes:
Method semen coicis to be measured is taken, test solution is prepared；
Semen coicis control medicinal material is followed the example of, reference substance solution is prepared；
Thin-layer chromatography: taking the test solution, reference substance solution, is put respectively on same lamellae, with volume ratio for (75
~82): (15~20): 1 petroleum ether, ether, glacial acetic acid mixture be that solvent is unfolded, it is aobvious to dry lamellae, spray
Toner, heating develop the color, and inspect under ultraviolet lamp.
2. the discrimination method of the method semen coicis according to claim 1 based on thin-layer chromatography, which is characterized in that the exhibition
It is (80~82): (15~18) that open agent, which be volume ratio: 1 petroleum ether, ether, glacial acetic acid mixture.
3. the discrimination method of the method semen coicis according to claim 2 based on thin-layer chromatography, which is characterized in that the exhibition
Open agent be volume ratio be 82:18:1 petroleum ether, ether, glacial acetic acid mixture.
4. the discrimination method of the method semen coicis according to any one of claims 1 to 3 based on thin-layer chromatography, feature exist
In the lamellae uses sodium carboxymethylcellulose for the silica gel g thin-layer plate of binder；The color developing agent is sulfuric acid and second
The mixture of alcohol, the volume of sulfuric acid accounts for 8%~12% in the mixture.
5. the discrimination method of the method semen coicis according to claim 4 based on thin-layer chromatography, which is characterized in that described is aobvious
Toner is the mixture of sulfuric acid and ethyl alcohol, and the volume of sulfuric acid accounts for 10% in the mixture.
6. the discrimination method of the method semen coicis according to any one of claims 1 to 3 based on thin-layer chromatography, feature exist
In described prepares test solution, includes the following steps:
Method semen coicis to be measured is taken, powder is made, the petroleum ether that temperature is 60 DEG C~90 DEG C is added, is ultrasonically treated, filtering, gained filter
Liquid is test solution；
The preparation reference substance solution, includes the following steps:
Semen coicis control medicinal material is followed the example of, powder is made, the petroleum ether that temperature is 60 DEG C~90 DEG C is added, is ultrasonically treated, filtering, institute
Obtaining filtrate is reference substance solution.
7. the discrimination method of the method semen coicis according to any one of claims 1 to 3 based on thin-layer chromatography, feature exist
In the temperature of the heating colour developing is 103~106 DEG C.
8. the discrimination method of the method semen coicis according to any one of claims 1 to 3 based on thin-layer chromatography, feature exist
In the ultraviolet light wavelength is 365nm.
9. the discrimination method of the method semen coicis according to any one of claims 1 to 3 based on thin-layer chromatography, feature exist
In the method semen coicis includes the method semen coicis medicine materical crude slice for the method that is prepared by the following procedure:
(1) clean semen coicis, boiling are taken；
(2) semen coicis obtained by step (1) is pulled out, draining is added edible oil, mixes thoroughly；
(3) semen coicis obtained by step (2) is steamed to the well-done heart, taking-up is dried, light shines；
(4) semen coicis obtained by sand stir-fry step (3) is flower-shaped to rice krispies, shakeout.
10. the discrimination method of the method semen coicis according to claim 9 based on thin-layer chromatography, which is characterized in that step (1)
In, the time of the boiling is 10~20min；In step (2), the edible oil is at least one of peanut oil, tea-seed oil；
2.5~the 2.8kg of edible oil is added in semen coicis obtained by every 100kg step (1)；Or/and the edible oil be selected from peanut oil,
At least one of tea-seed oil, castor oil.
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Citations (4)
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|---|---|---|---|---|
| CN101439172A (en) * | 2007-11-23 | 2009-05-27 | 北京亚东生物制药有限公司 | Chinese medicinal composition and preparation method thereof and quality control method |
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| CN103837636A (en) * | 2014-03-18 | 2014-06-04 | 安徽中医药大学第一附属医院 | HPTLC (high performance thin layer chromatography) quality evaluation method of Xinfeng capsule by adopting coix seeds as reference |
| CN103948839A (en) * | 2014-05-21 | 2014-07-30 | 徐葱茏 | Semen coicis prepared decoction pieces and preparation method thereof |
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2018
- 2018-09-13 CN CN201811067874.3A patent/CN108982741A/en active Pending
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|---|---|---|---|---|
| CN101439172A (en) * | 2007-11-23 | 2009-05-27 | 北京亚东生物制药有限公司 | Chinese medicinal composition and preparation method thereof and quality control method |
| CN101766771A (en) * | 2009-01-05 | 2010-07-07 | 王保安 | Quality control method of medicine for gynecopathy treatment |
| CN103837636A (en) * | 2014-03-18 | 2014-06-04 | 安徽中医药大学第一附属医院 | HPTLC (high performance thin layer chromatography) quality evaluation method of Xinfeng capsule by adopting coix seeds as reference |
| CN103948839A (en) * | 2014-05-21 | 2014-07-30 | 徐葱茏 | Semen coicis prepared decoction pieces and preparation method thereof |
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Application publication date: 20181211 |
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