CN108072731A - A kind of TLC Identification of Radix Glycyrrhizae hymsleya amabilis - Google Patents
A kind of TLC Identification of Radix Glycyrrhizae hymsleya amabilis Download PDFInfo
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- CN108072731A CN108072731A CN201611008719.5A CN201611008719A CN108072731A CN 108072731 A CN108072731 A CN 108072731A CN 201611008719 A CN201611008719 A CN 201611008719A CN 108072731 A CN108072731 A CN 108072731A
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Abstract
The invention discloses a kind of TLC Identifications of Radix Glycyrrhizae hymsleya amabilis, comprise the following steps：(1) preparation of test solution：Medicinal powder to be checked is taken, adds in methanol extraction, filtering is evaporated, methanol dissolved residue obtains test solution；(2) preparation of reference substance solution：Reference substance solution is prepared by reference substance of Hemsleyadin first；(3) thin layer differentiates：Respectively by reference substance solution and test solution on lamellae point sample, with chloroform acetone ethyl acetate volume ratio=7:5:1 is unfolded for solvent, and colour developing is inspected, comparison.The TLC Identification of the present invention, effectively each ingredient separation detection in Radix Glycyrrhizae hymsleya amabilis can be come out, active ingredient Hemsleyadin first especially therein, so as to quality that is accurate, simply and rapidly monitoring Radix Glycyrrhizae hymsleya amabilis, the quality control for Radix Glycyrrhizae hymsleya amabilis provides effective guarantee.
Description
Technical field
The present invention relates to medicinal material detection fields, and in particular to a kind of TLC Identification of Radix Glycyrrhizae hymsleya amabilis.
Background technology
Hymsleya amabilis is Curcurbitaceae hymsleya amabilis platymiscium dolichocarpel hemsleya root Hemsleya dolichocarpa W.J.Chang, high eyebrow is avenged
The colored hymsleya amabilis Hemsleya gigantha of courage Hemsleya omeiensis L.T.Shen et W.J.Chang or huge
The dried root of W.J.Chang.
After Radix Glycyrrhizae hymsleya amabilis is the bored profit 7.5h of succus liquiritiae with 0.4 times of amount of mass ratio, 80 DEG C dry the hymsleya amabilis dried after 12h
Product.Radix Glycyrrhizae hymsleya amabilis has clearing heat and detoxicating, and the effect of anti-inflammatory analgetic, for treating bacillary dysentery, enteritis, bronchitis is acute flat
Peach body is scorching, toothache caused by fire of deficiency type, the diseases such as stomachache.
Modern research shows that hymsleya amabilis is mainly used for treating mainly containing cucurbitacin and a small amount of dihydrocucurbitacin F and saponin constituent
The diseases such as bacillary dysentery, enteritis, bronchitis and acute tonsillitis.Wherein Hemsleyadin first content is higher, is its principle active component.
But at present still without it is a kind of can easy, Fast Monitoring Radix Glycyrrhizae hymsleya amabilis quality method.
The content of the invention
To solve the above problems, the present invention provides a kind of TLC Identification of Radix Glycyrrhizae hymsleya amabilis, including following
Step：
(1) preparation of test solution：
Medicinal powder to be checked is taken, adds in methanol extraction, filtering is evaporated, methanol dissolved residue obtains test solution；
(2) preparation of reference substance solution：
Reference substance solution is prepared by reference substance of Hemsleyadin first；
(3) thin layer differentiates：
Respectively by reference substance solution and test solution on lamellae point sample, with chloroform-acetone and ethyl acetate volume ratio
=7:5:1 is unfolded for solvent, and colour developing is inspected, comparison.
Further, in step (1), the extraction is ultrasound or refluxing extraction.
Further, in step (1), the extraction is ultrasonic extraction, when the time of ultrasonic extraction is 1 small.
Further, in the methanol extraction of the step (1), the envelope-bulk to weight ratio of methanol and medicinal material to be checked is 10mL:1g.
Further, in the methanol dissolved residue of the step (1), the envelope-bulk to weight ratio of methanol and medicinal material to be checked is
1.5mL:1g。
Further, the reference substance solution of the step (2) is the methanol solution of Hemsleyadin first；Preferably, Hemsleyadin first
Concentration be 1mg/mL.
Further, in step (3), the temperature of the expansion is 25 DEG C
Further, in step (3), the relative humidity of the expansion is 32%~72%.
Further, in step (3), the colour developing is using 5% phosphomolybdic acid ethanol solution as color developing agent, is added at 105 DEG C
Heat colour developing.
Further, the Radix Glycyrrhizae hymsleya amabilis is prepared by the following method：
It takes hymsleya amabilis medicine materical crude slice, adds in succus liquiritiae, mix thoroughly, bored profit 7.5h dries 12h at 80 DEG C, cools to obtain the final product；The hymsleya amabilis drink
The weight ratio of piece and Radix Glycyrrhizae is 5:2.
Experiment results proved, the present invention have been successfully established the TLC Identification of Radix Glycyrrhizae hymsleya amabilis, can effectively by
Each ingredient separation detection in Radix Glycyrrhizae hymsleya amabilis comes out, active ingredient Hemsleyadin first especially therein, so as to accurate, simple
Quality that is single, rapidly monitoring Radix Glycyrrhizae hymsleya amabilis, the quality control for Radix Glycyrrhizae hymsleya amabilis provide effective guarantee.
Obviously, the above according to the present invention according to the ordinary technical knowledge and customary means of this field, is not departing from
Under the premise of the above-mentioned basic fundamental thought of the present invention, the modification, replacement or change of other diversified forms can also be made.
The specific embodiment of form by the following examples remakes further specifically the above of the present invention
It is bright.But the scope that this should not be interpreted as to the above-mentioned theme of the present invention is only limitted to following example.It is all to be based on the above of the present invention
The technology realized all belongs to the scope of the present invention.
Description of the drawings
Fig. 1 is the thin-layer chromatogram of medicinal material to be checked and reference substance：1st, Hemsleyadin first reference substance；2~4, hymsleya amabilis sample processed.
Fig. 2 is investigated for extracting method under item, the thin-layer chromatogram of medicinal material and reference substance to be checked：1st, Hemsleyadin first reference substance；
2nd, methanol ultrasonic extraction；3rd, methanol is heated to reflux；4th, ethyl alcohol is heated to reflux；5th, EtOH Sonicate extracts.
Fig. 3 is solvent chloroform-ethanol volume ratio 19:When 1, the thin-layer chromatogram of medicinal material and reference substance to be checked：1st, hymsleya amabilis
Plain first reference substance；2~4, hymsleya amabilis sample processed.
Fig. 4 is solvent chloroform-methanol volume ratio=19:When 1, the thin-layer chromatogram of medicinal material and reference substance to be checked：1st, avenge
Choline first reference substance；2~4, hymsleya amabilis sample processed.
Fig. 5 is solvent chloroform-acetone and ethyl acetate volume ratio=5:3:When 1, the thin layer color of medicinal material and reference substance to be checked
Spectrogram：2nd, Hemsleyadin first reference substance；1st, 3,4, hymsleya amabilis sample processed.
Fig. 6 is solvent chloroform-acetone and ethyl acetate volume ratio=7:5:When 1, the thin layer color of medicinal material and reference substance to be checked
Spectrogram：1st, Hemsleyadin first reference substance；2~4, hymsleya amabilis sample processed.
When Fig. 7 is 30 DEG C of expansion of temperature, the thin-layer chromatogram of medicinal material and reference substance to be checked：1st, Hemsleyadin first reference substance；2~
4th, hymsleya amabilis sample processed.
When Fig. 8 is unfolded for 25 DEG C, the thin-layer chromatogram of medicinal material and reference substance to be checked：1st, Hemsleyadin first reference substance；2~4, make
Hymsleya amabilis sample.
When Fig. 9 is relative humidity 32%, the thin-layer chromatogram of medicinal material and reference substance to be checked：1st, Hemsleyadin first reference substance；2~
4th, hymsleya amabilis sample processed.
When Figure 10 is relative humidity 42%, the thin-layer chromatogram of medicinal material and reference substance to be checked：1st, Hemsleyadin first reference substance；2
~4, hymsleya amabilis sample processed.
When Figure 11 is relative humidity 47%, the thin-layer chromatogram of medicinal material and reference substance to be checked：2nd, Hemsleyadin first reference substance；1、
3rd, 4, hymsleya amabilis sample processed.
When Figure 12 is relative humidity 58%, the thin-layer chromatogram of medicinal material and reference substance to be checked：2nd, Hemsleyadin first reference substance；1、
3rd, 4, hymsleya amabilis sample processed.
When Figure 13 is relative humidity 65%, the thin-layer chromatogram of medicinal material and reference substance to be checked：1st, Hemsleyadin first reference substance；2
~4, hymsleya amabilis sample processed.
When Figure 14 is relative humidity 72%, the thin-layer chromatogram of medicinal material and reference substance to be checked：1st, Hemsleyadin first reference substance；2
~4, hymsleya amabilis sample processed.
When Figure 15 is relative humidity 88%, the thin-layer chromatogram of medicinal material and reference substance to be checked：1st, Hemsleyadin first reference substance；2
~4, hymsleya amabilis sample processed.
Specific embodiment
Experiment material used in following embodiments is as follows：
1 instrument and reagent
Instrument：Double flute expansion cylinder；Supersonic wave cleaning machine (model：SB-3200DT, the new limited public affairs of skill ultrasonic equipment in Ningbo
Department)；Vacuum drying chamber (DZF-6051, Shanghai will appoint experimental facilities Co., Ltd)；BP211D electronic balance (sensibility reciprocals
0.00001g, German Sartorius companies)；Gel GF 254 plate (10 × 20cm, Haiyang Chemical Plant, Qingdao)；Silica G plate is certainly
Making sheet.
Reagent：Cucurbitacin reference substance (lot number：111518-201103, purity 99.6%, Chinese food drug assay
Research institute).Water is distilled water, and other reagents are that analysis is pure.
Hymsleya amabilis is purchased from lotus pond market, is awarded through Lu elder generation of crude drug teaching and research room of pharmaceutical college of Chengdu University of Traditional Chinese Medicine penetrating judgment and is accredited as calabash
The dried root of Lu Ke plant hymsleya amabilis Hemsleya omeiensis L.T.Shen et W.J.Chang.
Hymsleya amabilis medicine materical crude slice：Above-mentioned hymsleya amabilis is taken, removes impurity, cleans, moisturizes.Sheet is cut, it is dry
It is prepared by 2 lamellaes
It prepares 0.3%CMC-Na solution left standstills to stay overnight, with silica G (proportioning about 3:1) mix, stir properly standby to viscosity
With.It is uniformly layered on clean glass plate, stands 12h, treats that plate dries in the shade after activating 0.5~1h at 110 DEG C, is placed in drying
It is spare in device.
The TLC Identification of 1 present invention of embodiment
This product powder 2g is taken, adds methanol 20mL, ultrasonic extraction 1h, is filtered, filtrate is put and is evaporated in water-bath, and residue adds methanol
3mL dissolves, as test solution.Hemsleyadin first reference substance is taken, adds methanol dissolving that solution of every 1mL containing 1mg is made, as right
According to product solution.According to thin-layered chromatography (《Chinese Pharmacopoeia》Four general rules 0502 of version in 2015) experiment, it is each to draw above two solution
10 μ l are put respectively on same silica gel g thin-layer plate, with chloroform-acetone and ethyl acetate (7:5:1) it is unfolded for solvent, takes out,
It dries, sprays the phosphomolybdic acid ethanol solution with 5%, it is clear to be heated to spot development at 105 DEG C.In test sample chromatography, with compareing
On the corresponding position of product chromatography, the spot of same color is shown.The results are shown in Figure 1.
The investigation of 2 the method for the present invention process conditions of embodiment
Under the conditions of the thin layer of embodiment 1, using single factor exploration method to extracting method, solvent, expansion temperature and exhibition
Humidity is opened to be investigated.
1st, the influence of different solvents, Different Extraction Method to hymsleya amabilis thin-layer chromatography processed
Using methanol, ethyl alcohol as extraction solvent, heating and refluxing extraction method, ultrasonic extraction Different Extraction Method and simultaneously are investigated
Investigate influence of the different solvents to hymsleya amabilis thin-layer chromatography processed.
4 parts of hymsleya amabilis powder 2g processed is weighed, is operated by table 1.
The different solvents and Different Extraction Method of 1 hymsleya amabilis processed of table
Filtration, filtrate are put and are evaporated in water-bath, and residue is dissolved respectively with methanol, ethyl alcohol, as test solution.Take Hemsleyadin
First reference substance adds methanol dissolving that solution of every 1mL containing 1mg is made, as reference substance solution.According to thin-layered chromatography (《Middle traditional Chinese medicines
Allusion quotation》Four general rules 0502 of version in 2015) experiment, each 10 μ l of above two solution are drawn, are put respectively in same silica gel g thin-layer plate
On, with chloroform-acetone and ethyl acetate (7:5:1) it is unfolded for solvent, takes out, dry, spray with 5% phosphomolybdic acid ethanol solution,
105 DEG C to be heated to spot development clear.In test sample chromatography, on position corresponding with reference substance chromatography, same color is shown
Spot.The results are shown in Figure 2.
As shown in Figure 2, the spot of methanol ultrasonic extraction can separate, and apparent, and ultrasonic extracting method is easy, easily grasp
Make, therefore selective extraction solvent and extracting method are methanol ultrasonic extraction.
2nd, the investigation of development system
To have selected chloroform-ethanol (19:1), chloroform-methanol (19:1), chloroform-acetone and ethyl acetate (5:3:1、7:5:
Etc. 1) several development systems are investigated.As a result as illustrated in figures 3-6.
From Fig. 3-6, development system chloroform-acetone and ethyl acetate (7:5:1) when being unfolded, maculiform in thin-layer chromatography
Shape is preferable, and separating degree is preferable, and Rf values are moderate, therefore it is selected to carry out indentification by TLC to hymsleya amabilis processed as solvent.
3rd, influence of the temperature to hymsleya amabilis thin-layer chromatography processed
Investigate 25 DEG C of room and 30 DEG C of influences to hymsleya amabilis thin-layer chromatography.When temperature is 30 DEG C, point of hymsleya amabilis thin-layer chromatography
Poor from spending, Rf values are larger, therefore select 25 DEG C of expansion.As a result as shown in Figure 7 and Figure 8.
4th, influence of the relative humidity to hymsleya amabilis thin-layer chromatography processed
The method for controlling relative humidity is：Expansion cylinder one end add in different proportion sulfuric acid, it is closed place 15~
30min adds solvent expansion.Reference table 2 prepares the sulfuric acid of different proportion.As a result as shown in Fig. 9-15.
Sulfuric acid ratio (V/V) needed for 2 different humidity of table
The results show that relative humidity, at 32%~72%, the influence unobvious to hymsleya amabilis thin-layer chromatography work as relative humidity
For 88% when, hymsleya amabilis thin-layer chromatography is had a significant impact, separating degree reduces, and spot is beyond recognition, therefore relative humidity control exists
32%~72%.
In conclusion the present invention has been successfully established the TLC Identification of Radix Glycyrrhizae hymsleya amabilis, it can be effectively by Radix Glycyrrhizae
Each ingredient separation detection in hymsleya amabilis processed comes out, active ingredient Hemsleyadin first especially therein, so as to it is accurate, simple,
The quality of Radix Glycyrrhizae hymsleya amabilis is rapidly monitored, the quality control for Radix Glycyrrhizae hymsleya amabilis provides effective guarantee.
Claims (10)
1. a kind of TLC Identification of Radix Glycyrrhizae hymsleya amabilis, it is characterised in that：Comprise the following steps：
(1) preparation of test solution：
Medicinal powder to be checked is taken, adds in methanol extraction, filtering is evaporated, methanol dissolved residue obtains test solution；
(2) preparation of reference substance solution：
Reference substance solution is prepared by reference substance of Hemsleyadin first；
(3) thin layer differentiates：
Respectively by reference substance solution and test solution on lamellae point sample, with chloroform-acetone and ethyl acetate volume ratio=7:
5:1 is unfolded for solvent, and colour developing is inspected, comparison.
2. TLC Identification according to claim 1, it is characterised in that：In step (1), the extraction is ultrasound
Or refluxing extraction.
3. TLC Identification according to claim 2, it is characterised in that：In step (1), the extraction is ultrasound
Extraction, when the time of ultrasonic extraction is 1 small.
4. according to claim 1-3 any one of them TLC Identifications, it is characterised in that：The first of the step (1)
In alcohol extracting, the envelope-bulk to weight ratio of methanol and medicinal material to be checked is 10mL:1g.
5. according to claim 1-4 any one of them TLC Identifications, it is characterised in that：The first of the step (1)
In alcohol dissolved residue, the envelope-bulk to weight ratio of methanol and medicinal material to be checked is 1.5mL:1g.
6. according to claim 1-4 any one of them TLC Identifications, it is characterised in that：Pair of the step (2)
According to the methanol solution that product solution is Hemsleyadin first；Preferably, the concentration of Hemsleyadin first is 1mg/mL.
7. according to claim 1-4 any one of them TLC Identifications, it is characterised in that：In step (3), the exhibition
The temperature opened is 25 DEG C.
8. according to claim 1-4 any one of them TLC Identifications, it is characterised in that：In step (3), the exhibition
The relative humidity opened is 32%~72%.
9. according to claim 1-4 any one of them TLC Identifications, it is characterised in that：It is described aobvious in step (3)
Color is using 5% phosphomolybdic acid ethanol solution as color developing agent, and colour developing is heated at 105 DEG C.
10. according to claim 1-9 any one of them TLC Identifications, it is characterised in that：The Radix Glycyrrhizae hymsleya amabilis
It is prepared by the following method：
It takes hymsleya amabilis medicine materical crude slice, adds in succus liquiritiae, mix thoroughly, bored profit 7.5h dries 12h at 80 DEG C, cools to obtain the final product；The hymsleya amabilis medicine materical crude slice with
The weight ratio of Radix Glycyrrhizae is 5:2.
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Cited By (1)
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| CN114814068A (en) * | 2022-05-13 | 2022-07-29 | 中山市中智药业集团有限公司 | Efficient thin-layer identification method for abrus cantoniensis hance and Abrus mollis |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN114814068A (en) * | 2022-05-13 | 2022-07-29 | 中山市中智药业集团有限公司 | Efficient thin-layer identification method for abrus cantoniensis hance and Abrus mollis |
| CN114814068B (en) * | 2022-05-13 | 2023-11-17 | 中山市中智药业集团有限公司 | Efficient thin-layer identification method for abrus herb and abrus herb |
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Application publication date: 20180525 |