CN104991031B - A kind of rhizoma smilacis glabrae medicinal material and the quick multi information thin-layer identification method of water extract thereof - Google Patents
A kind of rhizoma smilacis glabrae medicinal material and the quick multi information thin-layer identification method of water extract thereof Download PDFInfo
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Abstract
The present invention relates to the quick multi information thin-layer identification method of a kind of rhizoma smilacis glabrae medicinal material and water extract thereof.It is characterized in that: obtain test sample and control medicinal material solution with simple, fast pre-treating method, on same lamellae, 3 kinds different inspect under the conditions of inspected the multi information thin layer discriminating figure of rhizoma smilacis glabrae medicinal material and extract thereof, each clear spot, interlaced, but do not interfere with each other.Difference inspect under the conditions of multi information thin-layer chromatogram, not only increase the quality controllability of rhizoma smilacis glabrae medicinal material and water extract thereof, it is also the low cost of Rhizoma Smilacis Glabrae in noisy compound preparation, the discriminating of high efficiency quick thin layer, it is provided that the multi information condition of exclusive PCR selects.And method is easy, quick, high efficiency, low cost, non-environmental-pollution.
Description
Technical field
The present invention relates to the quick multi information thin-layer identification method of a kind of rhizoma smilacis glabrae medicinal material and water extract thereof.
Background technology
Chinese medicine Rhizoma Smilacis Glabrae has thin-layer identification method at present under the medical material item that Chinese Pharmacopoeia version in 2010 is recorded, but at its compound recipe
In preparation and water extract, the most it is not carried out the report of thin layer discriminating, consulting literatures【1、2】Although having thin in the formulation
The report that layer differentiates, but the most comparatively laborious complexity of pre-treatment of sample and control medicinal material, need by organic solvent purification process repeatedly,
The only pre-treatment of sample will take hours, the organic reagent of hundreds of ml, and efficiency is low, environmental pollution is serious, harm operator
Member's is healthy.Now find to obtain Rhizoma Smilacis Glabrae thin-layer identification method, enumerate and be analyzed as follows:
The thin-layer identification method of Chinese Pharmacopoeia one rhizoma smilacis glabrae medicinal material of version in 2010 takes this product powder 1g, adds methanol 20ml,
Supersound process 30 minutes, filters, and filtrate is as need testing solution.Separately take astilbin reference substance, add methanol and make every 1ml and contain
The solution of 0.1mg, as reference substance solution.Test according to thin layer chromatography (Chinese Pharmacopoeia one annex VIB of version in 2010),
Draw each 10 μ l of above two solution, put respectively on same silica gel g thin-layer plate, with toluene-ethyl acetate-formic acid (13:
32: 9) being developing solvent, launch, taking out, dry, spray, with aluminum chloride test solution, after placing 5 minutes, puts ultra-violet lamp (365nm)
Under inspect.In test sample chromatograph, on position corresponding with reference substance chromatograph, the speckle of aobvious same color.
This discrimination method be use astilbin reference substance be index, carry out Rhizoma Smilacis Glabrae thin layer discriminating, although before sample locate
Manage uncomplicated, but quantity of information is single, be only to inspect spraying aluminum chloride test solution, the fluorescence speckle after placing 5 minutes, differentiate one
Batch sample, sample pre-treatments spends 0.5 hour, organic solvent 20ml.
Document【1】In the Jieduxiezhu I granules of report, the thin-layer identification method of Rhizoma Smilacis Glabrae takes Jieduxiezhu I granules
In right amount, finely ground, weigh 2g, add methanol 50ml, supersound process 30 minutes, filter, filtrate is evaporated, and residue adds 3mol L-1
Hydrochloric acid 20ml, dissolves by several times, and is transferred in conical flask, and heating in water bath hydrolyzes 30 minutes, and taking-up lets cool, and adds chloroform 60ml,
Extracting in three times, combined chloroform layer, filter, be evaporated, residue adds methanol 2ml makes dissolving, as need testing solution.Separately take potato
Chinese yam sapogenin reference substance is appropriate, adds methanol and makes every 1ml solution containing 0.2mg, as reference substance solution.According to thin layer chromatography
(Chinese Pharmacopoeia one annex VIB of version in 2010) tests, and draws need testing solution 10 μ l, reference substance solution 5 μ l, point
Other point is on same silica gel g thin-layer plate, with chloroform-methanol (20: 0.2) as developing solvent, after presaturation 20 minutes, and exhibition
Opening, take out, dry, spray, with 10% phosphomolybdic acid ethanol solution, is heated to spot development at 105 DEG C clear.In test sample chromatograph,
On position corresponding with reference substance chromatograph, the speckle of aobvious same color.
This discrimination method be use diosgenin be index, carry out Rhizoma Smilacis Glabrae thin layer discriminating, differentiate a batch sample, only
3 hours sample pre-treatments time to be spent, organic solvent 230ml.
Document【2】In the Yinxie capsule of report, the thin layer of Rhizoma Smilacis Glabrae differentiates to take Yinxie capsule content 3g, and add water 60ml,
Supersound process 30 minutes, lets cool, and is transferred in separatory funnel, adds hydrochloric acid and adjusts pH to 2, adds ethyl acetate and shake extraction 2
Secondary, each 20ml, united extraction liquid, after being covered with the funnel of anhydrous sodium sulfate, filtrate is evaporated, adds methanol 1ml and make
Residue dissolves, and lysate is as need testing solution.Separately fetch earth Poria control medicinal material 5g, and add water 100ml, is heated to reflux 4 hours,
Filtering, filtrate is concentrated into about 10ml, adds ethanol 15ml, stirring, stands, and filters, and filtrate is evaporated, by the system of need testing solution
Preparation Method, prepares control medicinal material solution.Test according to thin layer chromatography (Chinese Pharmacopoeia one annex VIB of version in 2010), inhale
Take need testing solution and each 10 μ l of control medicinal material solution, put respectively on same silica gel g thin-layer plate, with chloroform-methanol-
Formic acid (15: 5: 1) is developing solvent, launches, and takes out, dries, and spray is with ferric chloride ethanol solution, in test sample chromatograph,
On position corresponding with control medicinal material chromatograph, the speckle of aobvious same color.
This discrimination method differentiates with Rhizoma Smilacis Glabrae control medicinal material for comparison thin layer, differentiates a batch sample, only sample and compares
The 7-8 hour time to be spent of pre-treatment of medical material, organic solvent 97ml.
Differentiated example by above-mentioned Rhizoma Smilacis Glabrae thin layer in the formulation, illustrate so far, its triviality differentiated, complexity,
Time-consuming, the unicity of quantity of information, to the contaminative of surrounding and high cost, poor efficiency.So raising detection efficiency,
Reduce testing cost, reduce environmental pollution, create rhizoma smilacis glabrae medicinal material and the quick multi information thin-layer identification method of water extract thereof,
Having become the instant objective of the struggle of testing staff, we are exactly under this background condition, invented rhizoma smilacis glabrae medicinal material and water carries
Take the quick multi information thin-layer identification method of thing.
The preparation method Rhizoma Smilacis Glabrae decoction pieces 7000-8000g of rhizoma smilacis glabrae medicinal material water extract or granule, adds water 8 respectively
Times amount, decoct extract secondary, each 1-2 hour, filter, 70 DEG C of concentrating under reduced pressure of filtrate, concentrated solution be spray-dried or 70 DEG C subtract
Press dry dry, obtain Rhizoma Smilacis Glabrae water extract dried cream powder；Or it is appropriate to add dextrin, mixing, pelletize, be respectively prepared 1000g, subpackage,
Obtain Rhizoma Smilacis Glabrae water extraction granule.
Summary of the invention
The present invention is aiming at Rhizoma Smilacis Glabrae water extract does not also have thin-layer identification method and its thin layer in compound preparation to differentiate
Loaded down with trivial details, time-consuming, quantity of information is single, method pollutes environment and the drawback of high cost, invented a kind of rhizoma smilacis glabrae medicinal material and water carried
Take the quick multi information thin-layer identification method of thing.That is, test sample and control medicinal material are obtained with the most simple, fast pre-treating method
Solution, on same lamellae, it is thus achieved that Rhizoma Smilacis Glabrae and its water extract are at ultra-violet lamp 365nm and 254nm, and spraying
10% ethanol solution of sulfuric acid colour developing after, daylight, 3 kinds inspect under the conditions of multi information thin layer discriminating figure, clear spot, interlaced,
But it does not interfere with each other (see Fig. 1,2,3).Compared with conventional single information thin layer, be not only greatly improved rhizoma smilacis glabrae medicinal material and
The quality controllability of its extract, beneficially quality surveillance, but also improve detection efficiency, saved cost, decrease ring
Environment pollution.And for extract compound preparation in Rhizoma Smilacis Glabrae quick thin layer differentiate, it is provided that three kinds inspect under the conditions of thin layer chromatography
Figure selects reference, in order to exclusive PCR composition.
The present invention solves the scheme that its technical problem used:
(1) thin layer of rhizoma smilacis glabrae medicinal material differentiates to take rhizoma smilacis glabrae medicinal material with its control medicinal material in right amount, finely ground, weighs 0.3g respectively,
Adding the 70% each 2ml of methanol, supersound process 20 minutes, supernatant is respectively as need testing solution and control medicinal material solution；In absorption
State control medicinal material solution and need testing solution each 5～8 μ l, put respectively in same silica gel G F254On lamellae, with volume ratio for 8:
Chloroform-acetate-methanol-the formic acid of 2: 3: 0.3 is developing solvent, launches, and takes out, hot blast drying, puts ultra-violet lamp
Inspect under 365nm, in test sample chromatograph, on position corresponding with control medicinal material chromatograph, aobvious same color fluorescence principal spot；
Put and inspect under ultra-violet lamp 254nm, in test sample chromatograph, on position corresponding with control medicinal material chromatograph, aobvious same color master
Speckle；Spraying with 10% ethanol solution of sulfuric acid again, 105 DEG C to be heated to spot development clear, in test sample chromatograph, with control medicinal material
On the corresponding position of chromatograph, the principal spot of aobvious same color.
(2) thin layer of Rhizoma Smilacis Glabrae water extract differentiates to take this product in right amount, finely ground, weighs 0.2g, adds 70% methanol 5ml,
Supersound process 10 minutes, filters, and filtrate is as need testing solution.Separately fetch earth Poria control medicinal material 0.3g, adds 70% methanol 2ml,
Supersound process 20 minutes, takes supernatant as control medicinal material solution.Test according to thin layer chromatography (annex VIB), in absorption
State control medicinal material solution and need testing solution each 5～8 μ l, put respectively in same silica gel G F254On lamellae, with volume ratio for 8:
Chloroform-acetate-methanol-the formic acid of 2: 3: 0.3 is developing solvent, launches, and takes out, hot blast drying, puts ultra-violet lamp
Inspect under 365nm, in test sample chromatograph, on position corresponding with control medicinal material chromatograph, aobvious same color fluorescence principal spot；
Put and inspect under ultra-violet lamp 254nm, in test sample chromatograph, on position corresponding with control medicinal material chromatograph, aobvious same color master
Speckle；Spraying with 10% ethanol solution of sulfuric acid again, 105 DEG C to be heated to spot development clear, in test sample chromatograph, with control medicinal material
On the corresponding position of chromatograph, the principal spot of aobvious same color.
The principle of the present invention is as follows:
Chemical constitution and character according to each effective ingredient of Chinese medicine, it then follows the extraction principle of similar compatibility, uses suitable proposing
Take solvent, easy, prepare test sample and control medicinal material solution quickly.Again by the movement of developing solvent, each composition is at lamellae
On the difference of the attached ability of Adsorption and desorption so that it is speckle is separated.Again by each compositional polarity size, choose polarity approximation
Effective ingredient, under the conditions of different inspecting, present different spot colors, on same lamellae, different inspects
Under the conditions of, it is thus achieved that the thin-layer chromatogram of multi information.
The innovative point of the present invention and having the beneficial effect that:
1. rhizoma smilacis glabrae medicinal material with the pre-treating method of control medicinal material is: weighs 0.3g rhizoma smilacis glabrae medicinal material respectively and compares medicine with it
Material, adds methanol 2ml, and after supersound process, supernatant is directly as test sample and control medicinal material solution；Easy, quick, only flower
Take about 20 minutes pre-treatment time, organic solvent 4ml.Compared with official method, although only saving process 10 minutes time,
Extraction solvent 16ml, but quantity of information is three times of report method.
2. the pre-treating method of Rhizoma Smilacis Glabrae water extract is also: take a small amount of Rhizoma Smilacis Glabrae water extract and control medicinal material, adds a small amount of
70% methanol, after supersound process, filtrate and supernatant are directly as test sample and control medicinal material solution；Spend the pre-treatment time altogether
20 minutes, organic solvent 7ml.Compared with report method, often detect a collection of medical material, save 2.5-7 hour time, You Jirong
Agent 90-223ml.
3. the quantity of information of detection is under the bright blue fluorescence principal spot under ultra-violet lamp (365nm), ultra-violet lamp (254nm)
2 fluorescence quenching speckles and colour developing after 3 colored speckles under daylight.Three kinds of differences inspect under the conditions of multi information collection of illustrative plates,
Condition of inspecting when providing exclusive PCR composition in compound preparation selects.
4. this discrimination method also differ in that with report method: the developing solvent of this discriminating is by chloroform, acetic acid
Ethyl ester, methanol, the organic solvent composition of 4 kinds of opposed polarities of formic acid, report method is by 2 kinds or 3 kinds of opposed polarities
Organic solvent forms, and considers from the separating resulting of similar compatibility, and the separating effect of developing solvent is proportionate with the kind of solvent, but
Obtain the difficulty of the optimal developing solvent component proportion kind also with solvent to be proportionate, thus through multiple developing solvent component with than
Example inquires into research, it is thus achieved that developing solvent best of breed, obtain three kinds inspect under the conditions of multi information collection of illustrative plates, and each information speckle
Separating effect is the most more satisfactory.
5. Rhizoma Smilacis Glabrae water extract is compared with the thin-layer identification method of Rhizoma Smilacis Glabrae in the preparation of report, and detection efficiency is high, detection
Low cost, non-environmental-pollution, for extract the quick thin layer of Rhizoma Smilacis Glabrae in compound preparation differentiate to provide three kinds inspect under the conditions of figure
Spectrum reference.
Accompanying drawing explanation
Fig. 1 is rhizoma smilacis glabrae medicinal material and extract thin layer TLC figure under ultra-violet lamp 365nm thereof.
Fig. 2 is rhizoma smilacis glabrae medicinal material and extract thin layer TLC figure under ultra-violet lamp 254nm thereof.
Thin layer TLC figure after Fig. 3 is rhizoma smilacis glabrae medicinal material and extract colour developing thereof, under daylight.
Fig. 1,2,3 are same lamellae, difference inspect under the conditions of thin-layer chromatogram, wherein, 1.2.3 is Rhizoma Smilacis Glabrae
Siberian cocklebur water extract；5.6.7 it is rhizoma smilacis glabrae medicinal material；4. it is Rhizoma Smilacis Glabrae control medicinal material
The specific embodiment of the invention is as follows:
(1) thin layer of rhizoma smilacis glabrae medicinal material differentiates to take rhizoma smilacis glabrae medicinal material with its control medicinal material in right amount, finely ground, weighs 0.3g respectively,
Adding the 70% each 2ml of methanol, supersound process 20 minutes, supernatant is respectively as need testing solution and control medicinal material solution；In absorption
State control medicinal material solution and need testing solution each 5～8 μ l, put respectively in same silica gel G F254On lamellae, with volume ratio for 8:
Chloroform-acetate-methanol-the formic acid of 2: 3: 0.3 is developing solvent, launches, and takes out, hot blast drying, puts ultra-violet lamp
Inspect under 365nm, in test sample chromatograph, on position corresponding with control medicinal material chromatograph, aobvious same color fluorescence principal spot；
Put and inspect under ultra-violet lamp 254nm, in test sample chromatograph, on position corresponding with control medicinal material chromatograph, aobvious same color master
Speckle；Spraying with 10% ethanol solution of sulfuric acid again, 105 DEG C to be heated to spot development clear, in test sample chromatograph, with control medicinal material
On the corresponding position of chromatograph, the principal spot of aobvious same color；
(2) thin layer of Rhizoma Smilacis Glabrae water extract differentiates to take this product in right amount, finely ground, weighs 0.2g, adds 70% methanol 5ml,
Supersound process 10 minutes, filters, and filtrate is as need testing solution.Separately fetch earth Poria control medicinal material 0.3g, adds 70% methanol 2ml,
Supersound process 20 minutes, takes supernatant as control medicinal material solution.Test according to thin layer chromatography (annex VIB), in absorption
State control medicinal material solution and need testing solution each 5～8 μ l, put respectively in same silica gel G F254On lamellae, with volume ratio for 8:
Chloroform-acetate-methanol-the formic acid of 2: 3: 0.3 is developing solvent, launches, and takes out, hot blast drying, puts ultra-violet lamp
Inspect under 365nm, in test sample chromatograph, on position corresponding with control medicinal material chromatograph, aobvious same color fluorescence principal spot；
Put and inspect under ultra-violet lamp 254nm, in test sample chromatograph, on position corresponding with control medicinal material chromatograph, aobvious same color master
Speckle；Spraying with 10% ethanol solution of sulfuric acid again, 105 DEG C to be heated to spot development clear, in test sample chromatograph, with control medicinal material
On the corresponding position of chromatograph, the principal spot of aobvious same color.
List of references
[1] Zhang Mingsheng, Gao Jiarong, Wei Liangbing " the indentification by TLC research of Jieduxiezhu I granules " Anhui medicine, 2007,11
(4)350-351
[2] Chen Xiuyuan, Li Hongyan, Zhao Hua " the thin layer chromatography qualitative identification of Yinxie capsule " Chinese Medicine science, 2013,3 (1)
111-112
Claims (2)
1. a rhizoma smilacis glabrae medicinal material and the quick multi information thin-layer identification method of water extract thereof, it is characterised in that:
(1) thin layer of rhizoma smilacis glabrae medicinal material differentiates to take rhizoma smilacis glabrae medicinal material with its control medicinal material in right amount, finely ground, weighs 0.3g respectively,
Adding the 70% each 2ml of methanol, supersound process 20 minutes, supernatant is respectively as need testing solution and control medicinal material solution；In absorption
State control medicinal material solution and need testing solution each 5～8 μ l, put respectively in same silica gel G F254On lamellae, with volume ratio for 8:
Chloroform-acetate-methanol-the formic acid of 2: 3: 0.3 is developing solvent, launches, and takes out, hot blast drying, puts ultra-violet lamp
Inspect under 365nm, in test sample chromatograph, on position corresponding with control medicinal material chromatograph, aobvious same color fluorescence principal spot；
Put and inspect under ultra-violet lamp 254nm, in test sample chromatograph, on position corresponding with control medicinal material chromatograph, aobvious same color master
Speckle；Spraying with 10% ethanol solution of sulfuric acid again, 105 DEG C to be heated to spot development clear, in test sample chromatograph, with control medicinal material
On the corresponding position of chromatograph, the principal spot of aobvious same color；
(2) thin layer of Rhizoma Smilacis Glabrae water extract differentiates to take this product in right amount, finely ground, weighs 0.2g, adds 70% methanol 5ml, super
Sonication 10 minutes, filters, and filtrate is as need testing solution；Separately fetch earth Poria control medicinal material 0.3g, adds 70% methanol 2ml,
Supersound process 20 minutes, takes supernatant as control medicinal material solution；Draw above-mentioned control medicinal material solution and need testing solution each 5～
8 μ l, put respectively in same silica gel G F254On lamellae, with chloroform-ethyl acetate that volume ratio is 8: 2: 3: 0.3-
Methyl alcohol-formic acid is developing solvent, launches, and takes out, hot blast drying, puts and inspect under ultra-violet lamp 365nm, in test sample chromatograph,
On position corresponding with control medicinal material chromatograph, aobvious same color fluorescence principal spot；Put and inspect under ultra-violet lamp 254nm, test sample
In chromatograph, on position corresponding with control medicinal material chromatograph, aobvious same color principal spot；Spray with 10% ethanol solution of sulfuric acid again,
105 DEG C are heated to spot development clearly, in test sample chromatograph, on position corresponding with control medicinal material chromatograph, aobvious same color
Principal spot.
A kind of rhizoma smilacis glabrae medicinal material the most according to claim 1 and the quick multi information thin-layer identification method of water extract thereof, its
It is characterised by the described Rhizoma Smilacis Glabrae water extract suitable primary crude drug of every 1g 7～8g.
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| CN111024879B (en) * | 2020-01-08 | 2021-08-31 | 河北中医学院 | Rapid multi-information thin-layer identification method for fresh motherwort herb freeze-dried powder, particles and standard decoction dry powder |
| CN112326862A (en) * | 2020-09-29 | 2021-02-05 | 鸿翔中药科技有限责任公司 | Poria cocos formula granules, preparation method thereof and thin-layer identification method |
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Inventor after: Niu Liying Inventor after: An Lina Inventor after: Wang Xinguo Inventor after: Han Guiru Inventor before: Wang Xinguo Inventor before: An Lina Inventor before: Niu Liying Inventor before: Han Guiru |
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Effective date of registration: 20201110 Address after: 050200 No. 3 Xingyuan Road, Luquan Economic Development Zone, Shijiazhuang City, Hebei Province Patentee after: HEBEI University OF CHINESE MEDICINE Patentee after: YUNNAN SHENWEI SHIPURUI PHARMACEUTICAL Co.,Ltd. Address before: 050200 No. 3 Apricot Garden Road, Luquan Economic Development Zone, Hebei, Shijiazhuang Patentee before: HEBEI University OF CHINESE MEDICINE |