CN104991030A - Radix scrophulariae medicinal material and rapid multi-information thin-layer identification method for water extract of radix scrophulariae medicinal material - Google Patents
Radix scrophulariae medicinal material and rapid multi-information thin-layer identification method for water extract of radix scrophulariae medicinal material Download PDFInfo
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Abstract
The invention relaets to a radix scrophulariae medicinal material and a rapid multi-information-thin-layer identification method for water extract of the radix scrophulariae medicinal material. The method is characterized in that test samples and control medicinal material solutions are obtained through a simple, convenient and rapid pretreatment method, on the same thin layer plate, the radix scrophulariae medicinal material and a multi-information-thin-layer identification diagram of the water extract of the radix scrophulariae medicinal material are inspected under three different inspection conditions, and all spots are clear and staggered with each other without interference. Compared with conventional single-information-thin-layer, the quality controllability of objects to be identified is improved greatly, quality supervision is facilitated, and the method is simple, convenient, rapid, high in efficiency, low in cost and free of environment pollution.
Description
Technical field
The present invention relates to the quick multi information thin-layer identification method of a kind of radix scrophulariae medicinal material and water extract thereof.
Background technology
The thin-layer identification method of Studying Chinese Crude Drug Figwort root and extraction preparation thereof, the pre-treatment of its sample and control medicinal material all compares very complicated, need by organic solvent purification process repeatedly, only the pre-treatment of sample will spend a few hours, the organic reagent of hundreds of ml, efficiency is low, environmental pollution is serious, endanger the healthy of operating personnel.For Chinese Pharmacopoeia version pharmacopeia in 2010, with regard to the thin-layer identification method of its medicinal material and its preparation, random extracts enumerates 3 examples:
1. radix scrophulariae single medicinal material TLC distinguish
Get this product powder 2g, add methyl alcohol 25ml, soak 1 hour, ultrasonic process 30 minutes, filter, filtrate evaporate to dryness, the residue 25ml that adds water makes dissolving, extracts 2 times, each 30ml with water saturated normal butyl alcohol, merge normal butyl alcohol liquid, evaporate to dryness, residue adds methyl alcohol 5ml makes dissolving, as need testing solution.Separately get radix scrophulariae control medicinal material 2g, be made in the same way of control medicinal material solution.Get harpagoside reference substance again, add methyl alcohol and make the solution of every 1ml containing 1mg, product solution in contrast.Test according to thin-layered chromatography (Chinese Pharmacopoeia version in 2010 annex VIB), draw each 4 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with methenyl choloride-methanol-water (12: 4: 1) lower floor solution for developping agent, put with in the developping agent presaturation expansion cylinder of 15 minutes, launch, take out, dry, spray with 5% vanillin-sulfuric acid solution, it is clear that hot blast blows to spot development.In test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the spot of aobvious same color.
This discrimination method differentiates a batch sample, spended time is wanted in sample and control medicinal material pre-treatment 3 hours, organic solvent 180ml.
2. radix scrophulariae TLC distinguish in preparation
A. radix scrophulariae TLC distinguish in Xuanmai Ganju grains
Get this product 10g, add water 40ml, and heating makes dissolving, lets cool, and filter, filtrate extracts 2 times with ether jolting, each 30ml, then extract with water saturated normal butyl alcohol 40ml jolting, and normal butyl alcohol liquid evaporate to dryness, residue adds methyl alcohol 1ml makes dissolving, as need testing solution.Separately get radix scrophulariae control medicinal material 2.5g, add ethanol 50ml, add hot reflux 1 hour, filter, filtrate is concentrated into dry, and the residue 30ml that adds water makes dissolving, is made in the same way of control medicinal material solution.Test according to thin-layered chromatography (Chinese Pharmacopoeia version in 2010 annex VIB), draw need testing solution 10 μ l, control medicinal material solution 6 μ l, put respectively on same silica gel g thin-layer plate, with methenyl choloride-methyl alcohol (5: 1) for developping agent, launch, take out, dry, spray is with 1% vanillin-sulfuric acid solution, and 105 DEG C to be heated to spot development clear.In test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the principal spot of aobvious same color.
This discrimination method differentiates a batch sample, spended time is wanted in sample and control medicinal material pre-treatment 3 hours, organic solvent 250ml.
B. radix scrophulariae TLC distinguish in gold fruit drink
Get this product 50ml, add ethyl acetate 50ml jolting to extract, the water saturated normal butyl alcohol of water intaking layer extracts 2 times (50ml, 30ml), merges normal butyl alcohol liquid, 2 (50ml are washed with ammonia solution, 30ml), then wash with normal butyl alcohol saturation water 50ml, get normal butyl alcohol liquid, evaporate to dryness, residue adds methyl alcohol 2ml makes dissolving, is added in neutral alumina column (100 ~ 200 orders, 2g, internal diameter is 1cm) on, with methyl alcohol 30ml wash-out, collect eluent, evaporate to dryness, residue adds absolute ethyl alcohol 0.5ml makes dissolving, as need testing solution.Separately get radix scrophulariae control medicinal material 2g, add water 50ml, adds hot reflux 1 hour, takes out, let cool, and filter, filtrate is made in the same way of control medicinal material solution.Test according to thin-layered chromatography (Chinese Pharmacopoeia version in 2010 annex VIB), draw each 20 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with methenyl choloride-methyl alcohol (10: 1) for developping agent, launch, take out, dry, spray with 5% vanillin-sulfuric acid solution.In test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the spot of aobvious same color.
This discrimination method differentiates a batch sample, spended time is wanted in sample and control medicinal material pre-treatment 3 hours, organic solvent 250ml.
By the TLC distinguish of above-mentioned radix scrophulariae single medicinal material and its TLC distinguish example in the formulation, confirm so far, the unicity of its triviality differentiated, complicacy, time-consuming, quantity of information, to the contaminative of surrounding environment and high cost, poor efficiency.So improve detection efficiency, reduction testing cost, minimizing environmental pollution, create the quick multi information thin-layer identification method of radix scrophulariae medicinal material and water extract thereof, become the instant objective of the struggle of testing staff, we are exactly under this background condition, have invented the quick multi information thin-layer identification method of radix scrophulariae medicinal material and water extract thereof.
Preparation method's radix scrophulariae medicine materical crude slice 4000-5000g of radix scrophulariae medicinal material water extract or granule, add water 8 times amount respectively, decocts and extract secondary, each 1-2 hour, filters, filtrate 70 DEG C of reduced pressure concentrations, concentrate spraying dry or 70 DEG C of drying under reduced pressure, obtain each water extract dried cream powder; Or it is appropriate to add dextrin, mixing, granulates, makes 1000g respectively, packing, obtain respective water extraction and get granule.
Summary of the invention
The present invention is exactly that TLC distinguish is loaded down with trivial details, time-consuming, quantity of information is single for radix scrophulariae medicinal material and in compound preparation, the drawback of method contaminated environment and high cost, has invented the quick multi information thin-layer identification method of a kind of radix scrophulariae medicinal material and water extract thereof.Namely, test sample and control medicinal material solution is obtained with the most simple, fast pre-treating method, on same thin layer plate, obtain radix scrophulariae and its water extract under ultraviolet lamp 365nm and 254nm and after 10% ethanol solution of sulfuric acid of spraying develops the color, daylight 3 kinds inspects the multi information TLC distinguish figure under condition, clear spot, interlaced, but do not interfere with each other (see Fig. 1,2,3).Compared with single information thin layer of routine, not only increase substantially the quality controllability of radix scrophulariae and extract thereof, be conducive to quality supervision, but also improve detection efficiency, saved cost, decrease environmental pollution.
The present invention solves the scheme that its technical matters adopts:
(1) TLC distinguish of radix scrophulariae medicinal material gets radix scrophulariae medicinal material and each 0.3g of control medicinal material, adds methyl alcohol 3ml respectively, ultrasonic process 15 minutes, and supernatant is respectively as test sample and control medicinal material solution; Draw need testing solution and each 5 ~ 6 μ l of control medicinal material solution, put respectively in same silica G F
254on thin layer plate, be that the chlorofonn-ethylacetate-methyl alcohol-dense ammonia of 4: 2: 4: 1 is for developping agent with volume ratio, launch, take out, dry, inspect under putting ultraviolet lamp 365nm and 254nm, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the fluorescence principal spot of aobvious same color and principal spot; Spray with volume ratio the mixed solution of the 5% vanillin-sulfuric acid solution-ethanol being 1: 6 again, 105 DEG C to be heated to spot development clear, inspects from the back side of thin layer plate, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, and the principal spot of aobvious same color;
(2) TLC distinguish of radix scrophulariae water extract gets radix scrophulariae water extract in right amount, and porphyrize, takes 0.3g, adds methyl alcohol 5ml, ultrasonic process 10 minutes, and filter, filtrate is as need testing solution; Separately get radix scrophulariae control medicinal material 0.3g, add methyl alcohol 3ml, ultrasonic process 15 minutes, supernatant is medicinal material solution in contrast; Draw need testing solution and each 5 ~ 6 μ l of control medicinal material solution, put respectively in same silica G F
254on thin layer plate, chlorofonn-ethylacetate-methyl alcohol-dense the ammonia being 4: 2: 4: 1 with volume ratio is that developping agent is for developping agent, launch, take out, dry, inspect under putting ultraviolet lamp 365nm and 254nm, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the fluorescence principal spot of aobvious same color and principal spot; Spray with volume ratio the mixed solution of the 5% vanillin-sulfuric acid solution-ethanol being 1: 6 again, 105 DEG C to be heated to spot development clear, inspects from the back side of thin layer plate, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, and the principal spot of aobvious same color;
Principle of the present invention is as follows:
According to chemical constitution and the character of Chinese medicine each effective constituent, follow the extraction principle of similar compatibility, adopt suitable Extraction solvent, easy, prepare test sample and control medicinal material solution quickly.Again by the movement of developping agent, the difference of the attached ability of the Adsorption and desorption of each composition on thin layer plate, makes its spot be separated.Again by each compositional polarity size, choose the effective constituent that polarity is approximate, under different conditions of inspecting, present different spot colors, on same thin layer plate, different inspect condition under, obtain the thin-layer chromatogram of multi information.
Innovative point of the present invention and beneficial effect as follows:
1. the pre-treating method of radix scrophulariae medicinal material is: get 0.3g radix scrophulariae medicinal material and control medicinal material, adds methyl alcohol 3ml, and after ultrasonic process, supernatant is directly as test sample and control medicinal material solution; Easy, quick, only about 15 minutes pre-treatment time of cost, organic solvent 6ml.Compared with report method, often detect a collection of medicinal material, save time 2.7 hours, organic solvent 174ml.
2. the pre-treating method of radix scrophulariae water extract also: take a morsel radix scrophulariae water extract and control medicinal material, adds proper amount of methanol, and after ultrasonic process, filtrate and supernatant are directly as test sample and control medicinal material solution; Spend 15 minutes pre-treatment time, organic solvent 8ml altogether.Compared with report method, often detect a collection of medicinal material, save time 2.7 hours, organic solvent 242ml.
3. the quantity of information detected is four kinds of colored speckles after the fluorescence spot under ultraviolet lamp (365nm), the sepia spot under ultraviolet lamp (254nm) and colour developing under daylight.These the three kinds information spots inspected under condition, analyze from its Rf value, be not same composition, more than its quantity of information, report method is also incomparable.
4. this discrimination method is also with the difference of report method: the developping agent of this discriminating is alkaline developers, and report method is slant acidity or neutral developping agent.This discriminating make use of the alkalescence of developping agent, can, by the material of some acidity, drag at initial point, make it not move with developping agent or move slowly, overcome the background interference of thin layer plate, realize sample and control medicinal material without purification process, also can obtain thin-layer chromatogram clearly.
5. the present invention is compared with the thin-layer identification method of report, improves detection efficiency 3 ~ 4 times, saves detection time 80%, reduces testing cost 80%, and reducing environmental pollution 90%, is an exemplary detection method with popularizing application prospect.
Accompanying drawing explanation
Fig. 1 is that radix scrophulariae medicinal material and the thin layer TLC of extract under ultraviolet lamp 254nm thereof scheme.
Fig. 2 is that radix scrophulariae medicinal material and the thin layer TLC of extract under ultraviolet lamp 365nm thereof scheme.
Fig. 3 is that after radix scrophulariae medicinal material and extract thereof develop the color, the thin layer TLC under daylight schemes.
Fig. 1,2,3 is same thin layer plate, and difference inspects the thin-layer chromatogram under condition, and wherein, 1.2.3 is radix scrophulariae water extract 5.6.7 is radix scrophulariae medicinal material; 4. be radix scrophulariae control medicinal material
The specific embodiment of the invention is as follows:
(1) TLC distinguish of radix scrophulariae medicinal material gets radix scrophulariae medicinal material and each 0.3g of control medicinal material, adds methyl alcohol 3ml respectively, ultrasonic process 15 minutes, and supernatant is respectively as test sample and control medicinal material solution; Draw need testing solution and each 5 ~ 6 μ l of control medicinal material solution, put respectively in same silica G F
254on thin layer plate, be that the chlorofonn-ethylacetate-methyl alcohol-dense ammonia of 4: 2: 4: 1 is for developping agent with volume ratio, launch, take out, dry, inspect under putting ultraviolet lamp 365nm and 254nm, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the fluorescence principal spot of aobvious same color and principal spot; Spray with volume ratio the mixed solution of the 5% vanillin-sulfuric acid solution-ethanol being 1: 6 again, 105 DEG C to be heated to spot development clear, inspects from the back side of thin layer plate, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, and the principal spot of aobvious same color.
(2) TLC distinguish of radix scrophulariae water extract gets radix scrophulariae water extract in right amount, and porphyrize, takes 0.3g, adds methyl alcohol 5ml, ultrasonic process 10 minutes, and filter, filtrate is as need testing solution; Separately get radix scrophulariae control medicinal material 0.3g, add methyl alcohol 3ml, ultrasonic process 15 minutes, supernatant is medicinal material solution in contrast; Draw need testing solution and each 5 ~ 6 μ l of control medicinal material solution, put respectively in same silica G F
254on thin layer plate, chlorofonn-ethylacetate-methyl alcohol-dense the ammonia being 4: 2: 4: 1 with volume ratio is that developping agent is for developping agent, launch, take out, dry, inspect under putting ultraviolet lamp 365nm and 254nm, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the fluorescence principal spot of aobvious same color and principal spot; Spray with volume ratio the mixed solution of the 5% vanillin-sulfuric acid solution-ethanol being 1: 6 again, 105 DEG C to be heated to spot development clear, inspects from the back side of thin layer plate, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, and the principal spot of aobvious same color.
Claims (3)
1. the present invention relates to the quick multi information thin-layer identification method of a kind of radix scrophulariae medicinal material and water extract thereof, it is characterized in that:
(1) TLC distinguish of radix scrophulariae medicinal material gets radix scrophulariae medicinal material and each 0.3g of control medicinal material, adds methyl alcohol 3ml respectively, ultrasonic process 15 minutes, and supernatant is respectively as test sample and control medicinal material solution; Draw need testing solution and each 5 ~ 6 μ l of control medicinal material solution, put respectively in same silica G F
254on thin layer plate, be that the chlorofonn-ethylacetate-methyl alcohol-dense ammonia of 4: 2: 4: 1 is for developping agent with volume ratio, launch, take out, dry, inspect under putting ultraviolet lamp 365nm and 254nm, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the fluorescence principal spot of aobvious same color and principal spot; Spray with volume ratio the mixed solution of the 5% vanillin-sulfuric acid solution-ethanol being 1: 6 again, 105 DEG C to be heated to spot development clear, inspects from the back side of thin layer plate, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, and the principal spot of aobvious same color;
(2) TLC distinguish of radix scrophulariae water extract gets radix scrophulariae water extract in right amount, and porphyrize, takes 0.3g, adds methyl alcohol 5ml, ultrasonic process 10 minutes, and filter, filtrate is as need testing solution; Separately get radix scrophulariae control medicinal material 0.3g, add methyl alcohol 3ml, ultrasonic process 15 minutes, supernatant is medicinal material solution in contrast; Draw need testing solution and each 5 ~ 6 μ l of control medicinal material solution, put respectively in same silica G F
254on thin layer plate, chlorofonn-ethylacetate-methyl alcohol-dense the ammonia being 4: 2: 4: 1 with volume ratio is that developping agent is for developping agent, launch, take out, dry, inspect under putting ultraviolet lamp 365nm and 254nm, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, the fluorescence principal spot of aobvious same color and principal spot; Spray with volume ratio the mixed solution of the 5% vanillin-sulfuric acid solution-ethanol being 1: 6 again, 105 DEG C to be heated to spot development clear, inspects from the back side of thin layer plate, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, and the principal spot of aobvious same color.
2. the quick multi information thin-layer identification method of a kind of radix scrophulariae medicinal material according to claim 1 and water extract thereof, is characterized in that the described suitable primary crude drug 4 ~ 5g of the every 1g of radix scrophulariae water extract.
3. the quick multi information thin-layer identification method of a kind of radix scrophulariae medicinal material according to claim 1 and water extract thereof, is characterized in that described water extract also can be the granule that its water extraction is got.
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