CN109030701B - Thin-layer identification method for quality control of radix stephaniae tetrandrae medicinal materials, decoction pieces and formula granules - Google Patents
Thin-layer identification method for quality control of radix stephaniae tetrandrae medicinal materials, decoction pieces and formula granules Download PDFInfo
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Abstract
The invention discloses a thin-layer identification method for stephania tetrandra medicinal materials, decoction pieces and formula granules, which comprises the following detection conditions: the stationary phase is silica gel GF254A thin layer plate; the developing solvent is toluene-ethyl acetate-methanolThe isopropanol-concentrated ammonia test solution has the volume ratio of (5-7) to (2-4) to (0.5-1.5) to (0.1-0.3). The invention can be used for controlling the quality of the stephania tetrandra medicinal materials and decoction pieces and can also be used for controlling the quality of the stephania tetrandra formula granules. The detection method has the characteristics of rich exclusive characteristic information, simplicity and convenience in operation, strong specificity, good repeatability and good application prospect.
Description
Technical Field
The invention relates to a thin-layer identification method for quality control of stephania tetrandra medicinal materials, decoction pieces and formula granules.
Background
The Stephania tetrandra is a dry root of Stephania tetrandra s.moore of a Stephania tetrandra plant of the family Menispermaceae, has the effects of dispelling wind, relieving pain, inducing diuresis and reducing edema, is clinically used for treating rheumatic arthralgia, edema, beriberi, dysuresia and eczema and sore toxin, and has a definite curative effect. Modern researches have shown that the active ingredient is mainly tetrandrine (C)38H42N2O6) And fangchinoline (C)37H40N2O6)。
In order to ensure the quality of the radix stephaniae tetrandrae, the quality control is required. At present, the thin-layer chromatography is adopted for identifying the stephania tetrandra medicinal material under the item of stephania tetrandra of the first edition of the 'Chinese pharmacopoeia' 2015 year edition, and the specific method is as follows:
taking 1g of the product powder, adding 15ml of ethanol, heating and refluxing for 1 hour, cooling, filtering, evaporating filtrate to dryness, and dissolving residue with 5ml of ethanol to obtain a sample solution. And mixing tetrandrine control and fangchinoline control with chloroform to obtain mixed solution containing 1mg of tetrandrine and fangchinoline control per 1ml as control solutions. Performing thin layer chromatography (general rule 0502) test, sucking 5 μ l of the above two solutions, respectively dropping on the same silica gel G thin layer plate, developing with chloroform-acetone-methanol-5% ammonia solution (6: 1:1: 0.1) as developing agent, taking out, air drying, and spraying with diluted bismuth potassium iodide solution. Spots of the same color appear in the chromatogram of the test solution at positions corresponding to those in the chromatogram of the control solution.
In the actual work of the identification method, because the identification spots are only two, the thin-layer chromatography information is relatively less, the specificity is low, the false positive result of sample application of a reference substance replacing a sample is easy to occur, the content of fangchinoline in the radix stephaniae tetrandrae is low, when the thin-layer chromatography condition is identified, the spots are seriously trailing and dispersed, so that the color development minimum limit of diluted bismuth potassium iodide is lower, the spots of fangchinoline in the thin-layer chromatography of the radix stephaniae tetrandrae sample cannot be observed, the false negative result occurs, the concentrated ammonia test solution in the developing agent is volatile, and the serious marginal effect can be generated when the saturation time of the developing agent is too short or too long, so that the identification of the radix stephaniae medicinal material is influenced.
Therefore, a method for identifying the stephania tetrandra medicinal materials, the decoction pieces and the formula granules, which has the advantages of more abundant exclusive characteristics, simple and easy operation, strong specificity and good repeatability, is required to be found.
Disclosure of Invention
In order to solve the problems, the invention provides a detection method for identifying characteristic components in stephania tetrandra medicinal materials, decoction pieces and formula granules.
The detection method provided by the invention adopts thin layer chromatography for detection, and comprises the following steps:
(1) preparation of control solutions: dissolving tetrandrine and fangchinoline in solvent to obtain control solution.
(2) Preparation of reference drug solution: extracting radix Stephaniae Tetrandrae control with solvent, filtering, evaporating filtrate, and dissolving residue with solvent to obtain control solution.
(3) Preparation of a test solution: taking the medicinal material of the stephania tetrandra, the decoction pieces of the stephania tetrandra or the formula particle powder of the stephania tetrandra, adding 5-15 times of ethanol, carrying out ultrasonic extraction for 20-40 min, cooling, filtering, evaporating filtrate to dryness, and adding 0.25-2.5 times of ethanol into residues to dissolve the residues to obtain a solution.
(4) The thin-layer chromatography identification method comprises the following steps: respectively dripping 5-10 μ l of reference solution, reference medicinal material solution, and sample solution on the same silica gel GF254Adding developing agent into the thin layer plate, saturating, adding the thin layer plate, continuously presaturating, developing, taking out, air drying, and inspecting under ultraviolet lamp (254 nm). In the chromatogram of the test solution, fluorescent spots of the same color appear at the corresponding positions of the chromatograms of the reference substance and the reference medicinal material.
Preferably, the conditions of each step of the thin layer chromatography detection are as follows:
(1) preparation of control solutions: taking a tetrandrine reference substance and a fangchinoline reference substance, and adding methanol to prepare mixed solutions containing 1-2 mg of tetrandrine and fangchinoline in each 1 ml.
(2) Preparation of reference drug solution: taking the tetrandra root reference medicinal material, adding 5-15 times of ethanol, carrying out ultrasonic extraction for 20-40 min, cooling, filtering, evaporating filtrate, and dissolving residues in 0.25-2.5 times of ethanol to obtain a solution.
(3) Preparation of a test solution: processing radix Stephaniae Tetrandrae medicinal material, decoction pieces or formula granules into fine powder, collecting powder, adding 4 times of ethanol, heating and refluxing for 30min, cooling, filtering, evaporating filtrate to dryness, and dissolving residue with 0.4 times of ethanol to obtain sample solution.
(4) The thin-layer chromatography identification method comprises the following steps: the developing solvent is toluene-ethyl acetate-methanol-isopropanol-concentrated ammonia test solution, and the volume ratio of the developing solvent to the concentrated ammonia test solution is (5-7): (2-4): (0.5-1.5): (0.1-0.3).
Further, it is preferable that the conditions of each step of the thin layer chromatography detection are as follows:
(1) preparation of control solutions: taking tetrandrine reference substance and fangchinoline reference substance, and adding methanol to obtain mixed solution containing 1mg of tetrandrine and fangchinoline in each 1 ml.
(2) Preparation of reference drug solution: collecting radix Stephaniae Tetrandrae control material, adding 4 times of ethanol, heating and refluxing for 30min, cooling, filtering, evaporating filtrate, and dissolving residue with 0.4 times of ethanol to obtain solution.
(3) Preparation of a test solution: processing radix Stephaniae Tetrandrae medicinal material, decoction pieces or formula granules into fine powder, collecting powder, adding 4 times of ethanol, heating and refluxing for 30min, cooling, filtering, evaporating filtrate to dryness, and dissolving residue with 0.4 times of ethanol to obtain solution.
(4) The thin-layer chromatography identification method comprises the following steps: the developing agent is toluene-ethyl acetate-methanol-isopropanol-concentrated ammonia test solution with the volume ratio of 6: 3: 1: 0.2, the solution is saturated for 10 minutes in an environment with the temperature of 4-35 ℃ and the relative humidity of 32-75%, and the solution is added into a thin layer plate and then is presaturated for 10 minutes.
The thin-layer identification method can be used as a method for detecting genuine stephania tetrandra medicinal materials and decoction pieces and controlling the quality of intermediate and finished products of stephania tetrandra formula granules, and has good practical value. The method can accurately detect and judge whether the medicinal materials and the decoction pieces of the fourstamen stephania root are genuine products carried by pharmacopoeia, can also accurately judge whether the feeding materials for producing the fourstamen stephania root formula particles are accurate, and can evaluate the condition of characteristic components of the fourstamen stephania root contained in particle intermediates or finished products so as to control the quality of the fourstamen stephania root formula particles.
Obviously, many modifications, substitutions, and variations are possible in light of the above teachings of the invention, without departing from the basic technical spirit of the invention, as defined by the following claims.
The present invention will be described in further detail with reference to the following examples. This should not be understood as limiting the scope of the above-described subject matter of the present invention to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.
Drawings
FIG. 1 shows the results of thin-layer chromatography detection of radix Stephaniae Tetrandrae;
FIG. 2 is a thin-layer chromatography detection result of radix Stephaniae Tetrandrae decoction pieces;
FIG. 3 is a thin-layer chromatography of radix Stephaniae Tetrandrae granules;
FIG. 4 shows a thin layer chromatogram developed by the development system I;
FIG. 5 thin layer chromatogram developed by development System II;
FIG. 6 thin layer chromatogram developed from development System III;
FIG. 7 thin layer chromatogram developed for development System IV;
FIG. 8 is a thin layer chromatogram developed using a cuttable plate available from Tianjin Silida technologies, Inc.;
FIG. 9 is a thin layer chromatogram developed using the Merck plate, Hencqi instruments, Inc. of Shanghai;
FIG. 10 is a thin layer chromatogram developed from a glass plate, which is a division of the Qingdao ocean plant;
FIG. 11 is a thin layer chromatogram developed in an environment at a temperature of 4 ℃;
FIG. 12 is a thin layer chromatogram developed in an environment at a temperature of 35 ℃;
FIG. 13 is a thin layer chromatogram developed in a humidity environment with a relative humidity of 32%;
FIG. 14 is a thin layer chromatogram developed in a humidity environment with a relative humidity of 75%;
FIG. 15 thin layer chromatograms for specificity study;
Detailed Description
Example 1 detection method of the invention
1. Detection method
(1) Preparation of control solutions: taking tetrandrine reference substance and fangchinoline reference substance, adding methanol to make into mixed solution containing 1mg of tetrandrine and fangchinoline respectively per 1ml, and using as reference substance solution.
(2) Preparation of reference drug solution: collecting radix Stephaniae Tetrandrae control material 2g, adding ethanol 10ml, ultrasonic extracting for 20min, cooling, filtering, evaporating filtrate, dissolving residue with ethanol 2ml to obtain control solution.
(3) Preparation of a test solution: taking 2g of radix Stephaniae Tetrandrae powder, adding 10ml of ethanol, performing ultrasonic extraction for 20min, cooling, filtering, evaporating filtrate to dryness, and dissolving residue with 2ml of ethanol to obtain a sample solution.
(4) The thin-layer chromatography identification method comprises the following steps: according to the thin-layer chromatography test of 0502 of the fourth general rule of the year 2015 of Chinese pharmacopoeia, 5 μ l of each of the reference solution, the reference solution and the sample solution is respectively spotted on the same silica gel GF254Adding toluene-ethyl acetate-methanol-isopropanol-concentrated ammonia test solution with the volume ratio of 5: 2: 0.5: 0.1 on a thin layer plate, spreading at the temperature of 4-35 ℃ and the relative humidity of 32-75 percentSaturating for 10 min, placing into thin layer plate, pre-saturating for 10 min, developing, taking out, air drying, and inspecting under ultraviolet lamp (254 nm). In the chromatogram of the test solution, fluorescent spots of the same color appear at the corresponding positions of the chromatograms of the reference solution and the reference material, as shown in FIG. 1.
Example 2 detection method of the invention
1. Detection method
(1) Preparation of control solutions: taking tetrandrine reference substance and fangchinoline reference substance, adding methanol to make into mixed solution containing 1.5mg of tetrandrine and fangchinoline respectively per 1ml as reference substance solution.
(2) Preparation of reference drug solution: collecting control radix Stephaniae Tetrandrae 5g, adding ethanol 20ml, ultrasonic extracting for 30min, cooling, filtering, evaporating filtrate, dissolving residue with ethanol 3ml to obtain control solution.
(3) Preparation of a test solution: taking 5g radix Stephaniae Tetrandrae decoction piece powder, adding 20ml ethanol, ultrasonic extracting for 30min, cooling, filtering, evaporating filtrate to dryness, and dissolving residue with 3ml ethanol to obtain sample solution.
(4) The thin-layer chromatography identification method comprises the following steps: according to the thin-layer chromatography test of 0502 of the fourth general rule of the 2015 th edition of Chinese pharmacopoeia, 5-10 mul of each of the reference solution, the reference solution and the sample solution are respectively spotted on the same silica gel GF254Adding toluene-ethyl acetate-methanol-isopropanol-concentrated ammonia test solution (the volume ratio is 6: 3: 1: 0.2) to the thin-layer plate, developing, saturating for 10 minutes in an environment with the temperature of 4-35 ℃ and the relative humidity of 32-75%, putting the thin-layer plate into the thin-layer plate, continuing presaturating for 10 minutes, developing, taking out, airing, and inspecting under an ultraviolet lamp (254 nm). In the chromatogram of the test solution, fluorescent spots of the same color appear at the corresponding positions of the chromatograms of the reference solution and the reference material, as shown in FIG. 2.
Example 3 detection method of the invention
1. Detection method
(1) Preparation of control solutions: taking tetrandrine reference substance and fangchinoline reference substance, adding methanol to make into mixed solution containing 1mg of tetrandrine and fangchinoline respectively per 1ml, and using as reference substance solution.
(2) Preparation of reference drug solution: collecting control radix Stephaniae Tetrandrae 5g, adding ethanol 20ml, ultrasonic extracting for 30min, cooling, filtering, evaporating filtrate, dissolving residue with ethanol 2ml to obtain control solution.
(3) Preparation of a test solution: taking 5g of radix Stephaniae Tetrandrae formula granule powder, adding 20ml of ethanol, performing ultrasonic extraction for 30min, cooling, filtering, evaporating filtrate to dryness, and dissolving residue with 2ml of ethanol to obtain sample solution.
(4) The thin-layer chromatography identification method comprises the following steps: according to the thin-layer chromatography test of 0502 of the fourth general rule of the 2015 th edition of Chinese pharmacopoeia, 5-10 mul of each of the reference solution, the reference solution and the sample solution are respectively spotted on the same silica gel GF254Adding toluene-ethyl acetate-methanol-isopropanol-concentrated ammonia test solution with the volume ratio of 6: 3: 1: 0.2 onto the thin-layer plate, developing, saturating for 10 minutes in an environment with the temperature of 4-35 ℃ and the relative humidity of 32-75%, putting the thin-layer plate into the thin-layer plate, continuing presaturating for 10 minutes, developing, taking out, airing, and inspecting under an ultraviolet lamp (254 nm). In the chromatogram of the test solution, fluorescent spots of the same color appear at the corresponding positions of the chromatograms of the reference solution and the reference material, as shown in FIG. 3.
Example 4 detection method of the invention
1. Detection method
(1) Preparation of control solutions: taking tetrandrine reference substance and fangchinoline reference substance, adding methanol to make into mixed solution containing 2mg of tetrandrine and fangchinoline respectively per 1ml as reference substance solution.
(2) Preparation of reference drug solution: collecting radix Stephaniae Tetrandrae control material 8g, adding ethanol 30ml, ultrasonic extracting for 40min, cooling, filtering, evaporating filtrate, dissolving residue with ethanol 5ml to obtain control solution.
(3) Preparation of a test solution: taking 8g of radix Stephaniae Tetrandrae formula granule powder, adding 30ml of ethanol, performing ultrasonic extraction for 40min, cooling, filtering, evaporating filtrate to dryness, and dissolving residue with 5ml of ethanol to obtain a sample solution.
(4) The thin-layer chromatography identification method comprises the following steps: according to the thin-layer chromatography test of 0502 of the fourth general rule of the 2015 version in Chinese pharmacopoeia, 5-10 mul of each of the reference solution, the reference medicinal material solution and the test solution is sucked,respectively spotted on the same silica gel GF254Adding toluene-ethyl acetate-methanol-isopropanol-concentrated ammonia test solution with the volume ratio of 7: 4: 1.5: 0.3 onto the thin-layer plate, developing, saturating for 10 minutes in an environment with the temperature of 4-35 ℃ and the relative humidity of 32-75%, placing the thin-layer plate into the thin-layer plate, continuing to presaturate for 10 minutes, developing, taking out, airing, and inspecting under an ultraviolet lamp (254 nm). In the chromatogram of the test solution, fluorescent spots of the same color appear at the corresponding positions of the chromatograms of the reference substance and the reference medicinal material.
Experimental example 1 examination of development System of detection method of the present invention
1. Instrument, reagent and experimental material
KQ5200DB model digital control ultrasonic cleaner (Kunshan ultrasonic Instrument Co., Ltd.); DK-98-II electrothermal constant temperature water bath (Qinst instruments, Tianjin Co., Ltd.), silica gel GF254Thin layer plate (Tianjin Silida science and technology Co., Ltd., cutting plate, batch number: 160122), silica gel GF254Thin layer plate (Qingdao Seawa silica gel desiccant Co., Ltd., glass plate, batch No. 20150502), silica gel GF254Thin layer plate (Qingdao ocean chemical plant division, glass plate, batch number: 20160611), thin layer chromatography scanner (CAMAG, made in Switzerland), automatic sample application instrument (CAMAG, made in Switzerland), tetrandrine control drug (China food and drug assay research institute, batch number: 121279-201502), tetrandrine control (China food and drug assay research institute, batch number: 110711-201609, purity 99.2%), tetrandrine control (China food and drug assay research institute, batch number: 110793-201606, purity 98.8%); the chemical reagents such as methanol, ethanol, toluene, ethyl acetate, isopropanol, acetone, chloroform, ammonia water and the like are analytically pure. Menispermum root prescription granule (batch number: 1803001, Sichuan new green pharmaceutical science and technology development Co., Ltd.)
2. Developer system inspection
(1) Preparation of a test solution: taking 5g of the powder, adding 20ml of ethanol, performing ultrasonic extraction for 30min, cooling, filtering, evaporating filtrate, dissolving residue with 2ml of ethanol to obtain a sample solution;
(2) preparation of reference drug solution: collecting control radix Stephaniae Tetrandrae 5g, adding ethanol 20ml, ultrasonic extracting for 30min, cooling, filtering, evaporating filtrate, dissolving residue with ethanol 2ml to obtain control solution;
(3) preparation of control solutions: taking tetrandrine reference substance and fangchinoline reference substance, adding methanol to make into mixed solution containing 1mg of tetrandrine and fangchinoline respectively per 1ml, and making into mixed reference substance solution.
(4) Thin-layer chromatography condition investigation: sucking 10 μ l of each of the reference solution, reference medicinal material solution and sample solution, and respectively dropping on the same silica gel GF254On the thin-layer plate, spreading systems I, II and III are respectively used.
Unfolding the system I: toluene-ethyl acetate-methanol-isopropanol-concentrated ammonia test solution (volume ratio of 6: 3: 1: 0.2)
And (3) unfolding a system II: toluene-ethyl acetate-methanol-formic acid (volume ratio 20: 10: 1)
Deployment system iii: toluene-Ethyl acetate-methanol-Water (volume ratio 2:4:2:1, upper layer solution placed below 10 ℃ C.)
And (4) unfolding the system IV: chloroform-acetone-methanol-5% concentrated ammonia test solution (volume ratio 6:1:1:0.1, Menispermaceae item lower thin layer development system of the national pharmacopoeia 2015 edition)
Developing with developing agent, and inspecting under ultraviolet lamp (254 nm).
3. The result of the detection
As shown in fig. 4-7, all component spots in the thin-layer chromatogram of the development system ii are at the front edge of the solvent, which indicates that the system cannot fully identify the components in the stephania tetrandra medicinal material; the spot tailing in the thin-layer chromatogram of the development system III is serious, and the fangchinoline and tetrandrine reference substances are not fully developed, so that fangchinoline medicinal materials cannot be fully identified; the IV thin-layer chromatography information of the development system is relatively less, the specificity is low, and false positive results of sample application by replacing samples with reference substances are easy to occur; the developing system I can fully separate the two reference substances, and compared with the developing systems II, III and IV, the developing system I has richer thin-layer chromatographic information and lower toxicity of reagents used by the system, so that the developing system I is selected as a developing system for thin-layer identification of the product.
Experimental example 2 durability examination of the detection method of the present invention
1. Instrument, reagent and experimental material
The conditions were the same as those in experimental example 1.
2. Method for investigating different thin-layer plates and results
Selecting silica gel GF of three different manufacturers254Thin layer plate, 10 μ l each of the control solution, the control solution and the sample solution in Experimental example 1, was spotted on the same silica gel GF254Developing the thin-layer plate according to the selected thin-layer chromatography conditions of the experimental example, taking out, airing, and inspecting under an ultraviolet lamp (254 nm).
As shown in FIGS. 8-10, under the selected thin layer chromatography conditions, different thin layer plates can achieve the purpose of identification.
3. Examination under different temperature conditions
Under the condition that the relative humidity is fixed at 60%, the temperature of the thin layer development environment is respectively changed, 10 mul of each of the reference solution, the reference medicinal material solution and the sample solution in the experimental example 1 are respectively taken, the samples are applied, the thin layer plates with the samples applied are taken, the thin layer plates are placed in the environment of 4 ℃ and 35 ℃ for development, taken out, dried and placed under an ultraviolet lamp (254nm) for inspection.
As shown in FIGS. 11-12, the thin layer chromatogram developed in the environment of 4-35 ℃ has clear main spots and good separation degree, and can achieve the identification purpose, so the thin layer identification method has good durability at 4-35 ℃.
4. Examination under different humidity conditions
As shown in FIGS. 13 to 14, under the condition that the temperature is fixed at 25 ℃, the relative humidity of the thin layer development environment is changed respectively, 10 μ l of each of the control solution, the control medicinal material solution and the sample solution in Experimental example 1 is sampled, the sampled thin layer plates are placed in the environment with the relative humidity of 32% and 75% respectively for development, taken out, dried and placed under an ultraviolet lamp (254nm) for inspection.
The result shows that the thin layer is spread in the environment with the relative humidity of 32 percent and 75 percent, the main spots are clear, the separation degree is good, and the method has good durability in the environment with the relative humidity of 32 percent to 75 percent.
Experimental example 3 investigation of specificity of the detection method of the present invention
1. Instrument, reagent and experimental material
The conditions were the same as those in experimental example 1.
2. Experimental methods
2.1 sample solution preparation method
(1) Preparing a test solution: grinding radix Stephaniae Tetrandrae formula granule, collecting powder 2g, adding ethanol 20ml, ultrasonic extracting for 30min, cooling, filtering, evaporating filtrate, and dissolving residue with ethanol 2ml to obtain sample solution;
(2) preparation of reference drug solution: collecting control radix Stephaniae Tetrandrae 5g, adding ethanol 20ml, ultrasonic extracting for 30min, cooling, filtering, evaporating filtrate, dissolving residue with ethanol 2ml to obtain control solution;
(3) preparation of control solutions: taking tetrandrine reference substance and fangchinoline reference substance, adding methanol to make into mixed solution containing 1mg of tetrandrine and fangchinoline respectively per 1ml, and making into mixed reference substance solution.
(4) Preparation of negative sample solution: preparing the final product without radix Stephaniae Tetrandrae according to the preparation process of radix Stephaniae Tetrandrae formula granule, grinding, collecting powder 2g, adding ethanol 20ml, ultrasonic extracting for 30min, cooling, filtering, evaporating filtrate, dissolving residue with ethanol 2ml to obtain negative sample solution.
2.2 thin layer specificity experiments
Taking 10 μ l of the control solution in step 3, 10 μ l of the control solution in step 2, and 10 μ l of each of the test solution in step 1 and the negative sample solution in step 4, and spotting on the same silica gel GF254On the thin layer plate, toluene-ethyl acetate-methanol-isopropanol-concentrated ammonia test solution with the volume ratio of 6: 3: 1: 0.2 is used as a developing agent, the developing agent is placed in a chromatographic cylinder, the thin layer plate is saturated for 10 minutes in the environment with the temperature of 25 ℃ and the relative humidity of 75 percent, then the thin layer plate is placed, the thin layer plate is placed in the chromatographic cylinder, the thin layer plate is presaturated for 10 minutes, the thin layer plate is developed, taken out and dried, the thin layer plate is inspected under ultraviolet light (254nm), and the observation is carried out, so.
3. Results of the experiment
As shown in fig. 15, the test sample without fangchinoline formulation granule did not interfere with the corresponding positions of the chromatography spots of fangchinoline and tetrandrine reference samples, indicating that the thin layer identification method has good specificity.
In conclusion, the thin-layer identification method can be used as a method for detecting genuine stephania tetrandra medicinal materials and decoction pieces and controlling the quality of intermediates and finished products of stephania tetrandra formula granules, and has good practical value. The method can accurately detect and judge whether the stephania tetrandra medicinal materials and decoction pieces are genuine products carried by pharmacopoeia, and can also accurately judge whether the feeding materials for the production of the stephania tetrandra formula particles are accurate, evaluate the condition of characteristic components contained in particle intermediates or finished products, and control the quality of the stephania tetrandra formula particles.
Claims (2)
1. A method for identifying traditional Chinese medicine radix stephaniae tetrandrae is characterized by comprising the following steps: identifying characteristic components of radix Stephaniae Tetrandrae, decoction pieces, formula granule intermediate product and formula granule finished product by thin layer chromatography; the method for identifying the characteristic components of the stephania tetrandra medicinal materials, the decoction pieces, the formula particle intermediate products and the formula particle finished products comprises the following steps:
(1) preparation of control solutions: taking tetrandrine reference substance and fangchinoline reference substance, adding methanol to make into mixed solution containing 1mg of tetrandrine and fangchinoline respectively per 1ml as reference substance solution;
(2) preparation of reference drug solution: taking a radix stephaniae tetrandrae reference medicinal material, adding 5-15 times of ethanol, carrying out ultrasonic extraction for 20-40 min, cooling, filtering, evaporating filtrate to dryness, and adding 0.25-2.5 times of ethanol into residues to dissolve the residues to obtain a solution serving as a reference medicinal material solution;
(3) preparation of a test solution: taking radix stephaniae tetrandrae medicinal materials, decoction pieces, formula particle intermediate products and formula particle finished product powder, adding 5-15 times of ethanol, carrying out ultrasonic extraction for 20-40 min, cooling, filtering, evaporating filtrate to dryness, and adding 0.25-2.5 times of ethanol into residues to dissolve the residues to obtain a solution serving as a test sample solution;
(4) the thin-layer chromatography identification method comprises the following steps: respectively dripping 5-10 μ l of reference solution, reference medicinal material solution, and sample solution on the same silica gel GF254Adding spreading agent into the thin layer plate, saturating, adding the thin layer plate, continuously presaturating, developing, taking out,air drying, placing under 254nm ultraviolet lamp, and observing to obtain fluorescent spots with the same color in the chromatogram of the sample and the corresponding positions of the chromatogram of the reference substance and the reference medicinal material;
the developing solvent is toluene-ethyl acetate-methanol-isopropanol-concentrated ammonia test solution, and the volume ratio of the developing solvent to the concentrated ammonia test solution is (5-7): (2-4): (0.5-1.5): (0.1-0.3).
2. The authentication method according to claim 1, wherein: the developing solvent in the step (4) is toluene-ethyl acetate-methanol-isopropanol-concentrated ammonia test solution with the volume ratio of 6: 3: 1: 0.2, the solution is saturated for 10 minutes in an environment with the temperature of 4-35 ℃ and the relative humidity of 32-75%, and is added with a thin layer plate and then presaturated for 10 minutes.
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| CN101028354B (en) * | 2006-03-01 | 2011-05-25 | 河北长天药业有限公司 | Detection method of siegesbeckia orientalis tablets |
| CN101371913A (en) * | 2008-09-08 | 2009-02-25 | 王衡新 | Quality control method of medicament for treatingcardiovascular and cerebrovascular diseases |
| CN102285994B (en) * | 2011-08-30 | 2013-08-14 | 聊城大学 | Method for separating and purifying fangchinoline and tetrandrine from stephania tetrandra |
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