CN110412198B - Detection method of traditional Chinese medicine composition for treating qi deficiency and blood stasis syndrome - Google Patents
Detection method of traditional Chinese medicine composition for treating qi deficiency and blood stasis syndrome Download PDFInfo
- Publication number
- CN110412198B CN110412198B CN201810328487.4A CN201810328487A CN110412198B CN 110412198 B CN110412198 B CN 110412198B CN 201810328487 A CN201810328487 A CN 201810328487A CN 110412198 B CN110412198 B CN 110412198B
- Authority
- CN
- China
- Prior art keywords
- solution
- methanol
- test
- ginsenoside
- thin
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/90—Plate chromatography, e.g. thin layer or paper chromatography
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention relates to a detection method of a traditional Chinese medicine composition for treating qi deficiency and blood stasis, wherein the traditional Chinese medicine composition is prepared from 500g of hedysarum polybotrys, 100g of pseudo-ginseng, 333g of rhizoma alismatis, 333g of angelica sinensis, 200g of ligusticum wallichii, 100g of cicada slough and 200g of radix curcumae and 100g of cassia twig; the detection method of the traditional Chinese medicine composition comprises the following thin-layer identification method of the medicinal materials of hedysarum polybotrys, cassia twig, ligusticum wallichii, angelica sinensis, pseudo-ginseng and rhizoma alismatis and the content determination method of notoginsenoside R1 and formononetin. The detection method has the characteristics of simple and convenient operation, reliable method and good reproducibility.
Description
Technical Field
The invention belongs to the field of detection of medicinal preparations, and particularly relates to a detection method of a traditional Chinese medicine composition for treating qi deficiency and blood stasis.
Background
The traditional Chinese medicine composition is qi-tonifying and blood circulation-promoting granules, the product is a unique variety owned by Shanxi step-size pharmaceutical Co Ltd, and the traditional Chinese medicine composition has a remarkable curative effect in treating cardiovascular and cerebrovascular diseases. The prescription of the traditional Chinese medicine composition comprises: 500g of radix hedysari, 100g of pseudo-ginseng, 333g of rhizoma alismatis, 333g of angelica, 200g of ligusticum wallichii, 100g of cicada slough, 200g of radix curcumae and 100g of cassia twig, and the preparation method comprises the following steps: the above eight ingredients are micronized and prepared into the pseudo-ginseng for later use. Soaking the rest seven materials of radix hedysari, rhizoma alismatis, angelica sinensis, ligusticum wallichii, cicada slough, radix curcumae, cassia twig and the like in 11 times of water, decocting for 1 hour, adding 10 times of water, decocting for two times, each time for 1 hour, filtering, combining filtrates, concentrating until the relative density is 1.20-1.23, adding a proper amount of pseudo-ginseng micro powder and dextrin, and preparing into 1000g particles. It has the functions of invigorating vital energy, promoting blood circulation, eliminating toxin and dredging meridian. The traditional Chinese medicine composition is mainly used for treating qi deficiency and blood stasis syndromes in traditional Chinese medicine, and symptoms of mental fatigue, weakness, short breath, no desire to speak, spontaneous perspiration, chest distress, limb numbness, pain, limb hemiplegia, amnesia, scaly skin, subcutaneous ecchymosis, dark tongue, enlarged tongue or tooth marks.
In order to further improve the production quality control and stability of the product, the invention aims to establish a qualitative and quantitative detection method which is strong in specificity, scientific, simple and convenient, and can ensure the quality and curative effect of the traditional Chinese medicine preparation.
Disclosure of Invention
The invention aims to provide a detection method of a traditional Chinese medicine composition for treating qi deficiency and blood stasis, which comprises the following thin-layer identification method and a method for measuring the contents of notoginsenoside R1 and formononetin. The technical scheme and the method have the characteristics of simplicity, feasibility, accuracy and good repeatability, and can be used as a quality control method of the traditional Chinese medicine composition to further ensure the stability and curative effect of the quality of a medicine product.
In the quality standard draft research, the traditional Chinese medicine composition is finally determined after a large number of tests and repeated groping.
In the formula, radix hedysari is the main medicinal material of the formula, and formononetin is the formononetinThe main effective components of the medicine are used for content determination by taking formononetin as an index, and the quality of a finished product is controlled. Reference and repeated experiments prove that the high performance liquid chromatography is adopted, so that the operation is simpler and more convenient, the method is reliable, and the reproducibility is good, so that the high performance liquid chromatography is adopted in the standard to measure the content of the formononetin. In addition, the content of another Chinese medicinal material of notoginseng in the prescription is controlled, and the notoginsenoside R in the dried product is controlled1(C47H80O18) Ginsenoside Rg1(C42H72O14) Ginsenoside Rb1(C54H92O23) The total amount of (A) is limited.
The reference medicinal materials or the reference substances are used as the standard, thin-layer chromatography identification is carried out on the radix hedysari, the ligusticum wallichii, the rhizoma alismatis, the pseudo-ginseng and the like in the finished product, and the reproducibility is better through three batches of pilot samples and mass production sample inspection and is listed under the quality standard identification item. When the periostracum cicadae and the curcuma aromatica in the formula are subjected to thin-layer qualitative inspection, a large number of tests show that negative test sample solutions of the cicada slough and the curcuma aromatica have interference and cannot be detected, so that inspection items are not listed in the quality standard.
The technical scheme of the patent application of the invention is as follows:
the detection method of the traditional Chinese medicine composition comprises the following thin-layer identification method and a method for measuring the contents of notoginsenoside R1 and formononetin.
The thin layer identification method of the detection method comprises the following steps:
(1) the method for identifying the radix hedysari comprises the following steps: grinding the Chinese medicinal composition, adding methanol, filtering, evaporating filtrate to dryness, dissolving residue in water, extracting with diethyl ether, evaporating to dryness, and dissolving residue to obtain sample solution; adding methanol into formononetin reference substance to obtain reference substance solution; and (3) performing thin-layer chromatography test, namely respectively dropping the two solutions on the same silica gel thin-layer plate according to the ratio of petroleum ether to ethyl acetate of 0.5-2: 0.5-2 of developing agent, presaturating with developing agent, developing, taking out, drying, and inspecting under 254nm ultraviolet lamp to obtain spots with same color in the chromatogram of the test sample and the corresponding positions of the chromatogram of the reference sample;
(2) the method for identifying the cassia twig comprises the following steps: taking a cassia twig reference medicinal material, preparing a reference medicinal material solution according to the preparation method of the test solution under the item of the identification (1), and adding methanol into a cinnamic acid reference substance to prepare a solution serving as a reference substance solution; and (2) performing thin-layer chromatography test, namely absorbing the sample solution and the two solutions under the identification item (1), respectively dropping the sample solution and the two solutions on the same silica gel thin-layer plate, and performing thin-layer chromatography test on the sample solution by using petroleum ether-n-hexane-ethyl formate-formic acid 1.5-2.5: 5-7: 2-4: 0.1-0.3 of developing agent, developing, taking out, drying, and inspecting under an ultraviolet lamp at 254nm, wherein spots with the same color appear in the chromatogram of the test solution at the positions corresponding to the chromatograms of the reference medicinal material and the reference solution;
(3) the identification method of the ligusticum wallichii comprises the following steps: taking rhizoma Ligustici Chuanxiong as reference material, and making into reference material solution according to the preparation method of the sample solution under item (1). And (2) performing thin-layer chromatography test, namely absorbing the sample solution and the reference medicinal material solution under the item (1) for identification, respectively dropping the sample solution and the reference medicinal material solution on the same silica gel thin-layer plate, and performing thin-layer chromatography test on the sample solution and the reference medicinal material solution by using n-hexane-ethyl acetate 0.8-1.2: 0.8-1.2 of developing agent, developing, taking out, drying, placing under an ultraviolet lamp at 365nm for inspection, and displaying fluorescent spots with the same color in the chromatogram of the test solution at the position corresponding to the chromatogram of the reference solution;
(4) the identification method of the angelica comprises the following steps: adding methanol into ferulic acid control to obtain control solution. And (2) performing thin-layer chromatography test, namely sucking the test solution and the reference solution under the identification item (1), respectively dropping the test solution and the reference solution on the same silica gel thin-layer plate, and adding 3.5-4.5 parts of cyclohexane-trichloromethane-ethyl acetate-formic acid: 0.8-1.2: 3.5-4.5: 0.05-0.15 of developing agent, developing, taking out, airing, and placing under an ultraviolet lamp at 254nm for inspection, wherein spots with the same color appear in the chromatogram of the test solution at the positions corresponding to the chromatograms of the reference solution;
(5) the identification method of the pseudo-ginseng comprises the following steps: grinding the Chinese medicinal composition, adding methanol, filtering, evaporating the filtrate to dryness, dissolving the residue in water, extracting with saturated n-butanol, washing with ammonia solution, evaporating the n-butanol solution to dryness, dissolving the residue in methanol to obtain test solution, and collecting ginsenoside Rb1Reference substance, ginsenoside Re reference substance, and notoginsenoside R1Reference substance and ginsenoside Rg1A reference substance is prepared by the following steps of,adding methanol to prepare a mixed solution containing 0.5mg per 1ml, taking the mixed solution as a reference solution, performing thin layer chromatography test, sucking the two solutions, respectively dropping the two solutions on the same silica gel thin layer plate, and mixing the two solutions with chloroform-methanol-water in a ratio of 60-70: 32-37: 8-12, taking out, airing, spraying a 10% sulfuric acid ethanol solution, heating at 105 ℃ until spots are clear, wherein spots with the same color appear in the chromatogram of the test sample at the position corresponding to the chromatogram of the reference sample;
(6) the identification method of the rhizoma alismatis comprises the following steps: grinding the Chinese medicinal composition, adding methanol, ultrasonic treating, filtering, evaporating filtrate to dryness, dissolving residue in water, extracting with ethyl acetate, evaporating to dryness, and dissolving residue in methanol to obtain sample solution; adding ethyl acetate into rhizoma alismatis reference medicinal material, performing ultrasonic treatment, filtering, evaporating filtrate to dryness, dissolving residue by adding methanol to obtain reference medicinal material solution, performing thin-layer chromatography test, sucking the two solutions, respectively dropping the two solutions on the same silica gel thin-layer plate, and adding cyclohexane-ethyl acetate 0.8-1.2: 0.8-1.2 of developing agent, developing, taking out, drying, spraying 5% silicotungstic acid ethanol solution, heating at 105 ℃ until the spots are clear, and showing spots with the same color in the chromatogram of the test solution at the position corresponding to the chromatogram of the reference medicinal material.
Notoginsenoside R in the Chinese medicinal composition1The content determination method comprises the following steps:
the method comprises the following steps of: acetonitrile is used as a mobile phase A, water is used as a mobile phase B, and gradient elution is carried out according to the following elution proportion: time, 0-20 minutes, mobile phase a (%): 21 → 50, mobile phase B (%) 79 → 50;
preparing a control solution: precisely weighing notoginsenoside R1Reference substance, ginsenoside Rg1Reference substance and ginsenoside Rb1Adding methanol to a control sample to obtain a solution containing notoginsenoside R per 1ml10.1-0.3 mg of ginsenoside Rg10.4-0.6 mg of ginsenoside Rb10.3-0.5 mg of mixed solution to obtain,
preparing a test sample solution: mixing the Chinese medicinal composition, grinding, accurately weighing, adding methanol, weighing, refluxing in water bath for 1-3 hr, cooling, weighing, adding methanol to balance weight, shaking, filtering, collecting filtrate,
fourth, measurement method: precisely sucking the reference solution and the sample solution, respectively, injecting into a liquid chromatograph, and measuring.
The method for measuring the content of formononetin in the traditional Chinese medicine composition comprises the following steps:
the method comprises the following steps of: methanol-water 58:42 is used as a mobile phase; the detection wavelength is 250nm, and the column temperature is 25-35 ℃;
preparing a control solution: taking a proper amount of formononetin reference substances, precisely weighing, and adding methanol to prepare a solution containing 6-10 mu g of formononetin per 1ml to obtain the formononetin reference substances;
preparing a test sample solution: mixing the Chinese medicinal composition, grinding, accurately weighing, placing in a conical flask, adding methanol, refluxing in water bath, filtering, placing the residue and filter paper in the conical flask, adding methanol, refluxing, mixing the filtrates, washing the filter paper with methanol, concentrating under reduced pressure, metering the volume of the concentrated solution to a measuring flask, shaking, filtering, and collecting the filtrate.
The technical scheme of the invention has the following beneficial effects:
1. the detection method establishes a thin-layer chromatography identification project, and qualitatively identifies medicinal materials or characteristic components of ligusticum wallichii, cassia twig, radix hedysari, pseudo-ginseng, rhizoma alismatis and angelica sinensis in a prescription composition. One of the biggest characteristics is: the preparation methods of the sample solutions adopted by 4 of the 6 identifications are consistent, great convenience is brought to the qualitative identification of the variety, the production cost is reduced in the quality control link, the qualitative identification is carried out, and the established method has the advantages of high separation degree, good reproducibility and strong specificity.
2. Radix hedysari is the monarch drug in the formula of the medicine, and formononetin is the main effective component of the medicine, so the content determination is carried out by taking the formononetin as an index, and the quality of a finished product is controlled. Notoginseng radix is a valuable medicine, so its effective components are also quality controlled. Therefore, the content of the saponin active ingredients in the formononetin and the pseudo-ginseng is measured by adopting a high performance liquid chromatography in the standard.
The experimental result shows that the notoginseng soapGlycoside R1Ginsenoside Rg1And ginsenoside Rb1The precision test of (A) shows that the notoginsenoside R1Ginsenoside Rg1And ginsenoside Rb1RSD of the peak area natural logarithm is less than or equal to 2 percent, and the sample solution is basically stable within 24 hours; the average recovery results were: the content of notoginsenoside R1 is 99.79%; ginsenoside Rg1 is 98.34%; ginsenoside Rb1 was 100.5% with a relative standard deviation of 3.0%. The method for measuring the content of the formononetin is also established, and the precision test of the method is that the RSD is less than or equal to 0.22 percent, the repeatability test is carried out, the RSD is less than or equal to 1.87 percent, the stability test is carried out, the RSD is less than or equal to 3.14 percent, the sample adding and recycling test is carried out, and the RSD is less than or equal to 1.17 percent; the experimental data show that the detection method established by the application has the characteristics of simple and convenient operation, reliable method and good reproducibility.
3. The application establishes a detection method of multi-index components, and finally determines that each bag of the product contains panax notoginseng and ginsenoside Rg1(C42H72O14) Ginsenoside Rb1(C54H92O23) And notoginsenoside R1(C47H80O18) Not less than 50mg, based on the total amount of (A) and (B). The further control of the effective components ensures the inherent quality and curative effect of the product.
Drawings
FIG. 1-thin-layer chromatography chromatogram for radix Hedysari identification, wherein 1-lot number of test solution-01, 2-lot number of test solution-02, 3-formononetin reference, 4-lot number of test solution-03;
FIG. 2-thin layer chromatogram for identification of ramulus Cinnamomi, wherein 1-test solution lot-01, 2-test solution lot-02, 3-ramulus Cinnamomi reference medicinal material, 4-test solution lot-03;
FIG. 3-thin layer chromatogram for identification of rhizoma Ligustici Chuanxiong, wherein 1-test solution lot-01, 2-test solution lot-02, 3-rhizoma Ligustici Chuanxiong reference medicinal material, 4-test solution lot-03;
FIG. 4-thin layer chromatogram for identification of Angelica sinensis, wherein 1-lot number of test solution-01, 2-lot number of test solution-02, 3-ferulic acid reference, 4-lot number of test solution-03;
FIG. 5-thin layer chromatogram for identification of Notoginseng radix, wherein 1-test solution lot-01, 2-test solution lot-02, 3-Ginseng radix and notoginsenoside mixed reference, 4-test solution lot-03;
FIG. 6-thin-layer chromatography for identification of Alismatis rhizoma, wherein 1-test sample solution lot-01, 2-test sample solution lot-02, 3-Alismatis rhizoma control medicinal material, 4-test sample solution lot-03;
FIG. 7 shows chromatogram of blank experiment and reference solution of 1-notoginsenoside R1, 2-ginsenoside Rg1, and 3-ginsenoside Rb 1.
FIG. 8-formononetin blank experiment and control solution chromatograms.
Detailed Description
Unless defined otherwise, technical or scientific terms used herein in the specification and claims of the present patent application shall have the ordinary meaning as understood by those of ordinary skill in the art to which the present invention belongs. The Chinese medicinal composition is qi-tonifying and collateral-dredging granules, and the prescription proportion and the preparation method of the Chinese medicinal composition have definite meanings in the content part of the invention.
Example 1
The thin-layer identification method of radix hedysari comprises the following steps:
(1) taking 10g of the product, grinding, adding 30ml of methanol, performing ultrasonic treatment for 30 minutes, filtering, and evaporating the filtrate to dryness. Dissolving the residue with 20ml water, transferring into separating funnel, extracting with diethyl ether for 3 times (20 ml each time), mixing diethyl ether solutions, evaporating to dryness, and dissolving the residue with 1ml methanol to obtain sample solution. Taking formononetin reference substance, adding methanol to make into solution containing 0.1mg per 1ml as reference substance solution. Subjecting to thin layer chromatography (general rule 0502), sucking 5 μ l of the above two solutions, and spotting on the same silica gel GF254And (3) taking petroleum ether (30-60 ℃) and ethyl acetate (1:1) as a developing agent on the thin-layer plate, placing the thin-layer plate in a developing cylinder pre-saturated with the developing agent for 20 minutes, developing, taking out, airing, and inspecting under an ultraviolet lamp (254 nm). Spots of the same color appear in the chromatogram of the test solution at the positions corresponding to those in the chromatogram of the control solution.
Example 2
The cassia twig thin layer identification method comprises the following steps:
taking 0.5g of cassia twig as a reference medicinal material, and preparing the reference medicinal material solution according to the preparation method of the test solution under the item of the identification (1). Taking cinnamic acid reference substance, adding methanol to make into solution containing 0.5mg per 1ml as reference substance solution. Performing thin layer chromatography (general rule 0502) test by collecting 3 μ l of each of the sample solution and the above two solutions under (1) and spotting on the same silica gel GF254And (3) developing the thin-layer plate by using petroleum ether (30-60 ℃) -n-hexane-ethyl formate-formic acid (2:6:3:0.2) as a developing agent, taking out, airing, and inspecting under an ultraviolet lamp (254 nm). Spots of the same color appear on the chromatogram of the test solution at the positions corresponding to the chromatograms of the control solution and the reference solution.
Example 3
The thin layer identification method of the ligusticum wallichii comprises the following steps:
taking 0.5g of rhizoma Ligustici Chuanxiong as reference material, and making into reference material solution according to the preparation method of the test solution under item (1). Performing thin layer chromatography (general rule 0502) test, collecting 2 μ l of each of the sample solution and the control solution under the item (1), respectively dropping on the same silica gel G thin layer plate, developing with n-hexane-ethyl acetate (1:1) as developing agent, taking out, air drying, and inspecting under ultraviolet lamp (365 nm). In the chromatogram of the test solution, fluorescent spots with the same color appear at the corresponding positions of the chromatogram of the reference solution.
Example 4
The angelica sinensis thin-layer identification method comprises the following steps:
taking ferulic acid control, adding methanol to make 1ml solution containing 1mg as control solution. Performing thin layer chromatography (general rule 0502) test by sucking 1 μ l of each of the sample solution and the above control solution under item (1), and spotting on the same silica gel GF254Spreading on thin layer plate with cyclohexane-chloroform-ethyl acetate-formic acid (4:1:4:0.1) as developing agent, taking out, air drying, and inspecting under ultraviolet lamp (254 nm). 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.
Example 5
The thin-layer pseudo-ginseng identification method comprises the following steps:
take 5g of this herb and grind it into fine powderAdding 30ml of methanol, performing ultrasonic treatment for 30 minutes, filtering, and evaporating the filtrate to dryness. Dissolving the residue with 10ml water, transferring into separating funnel, extracting with water saturated n-butanol for 3 times (10 ml each time), mixing n-butanol solutions, washing with ammonia solution for 3 times (10 ml each time), evaporating n-butanol solution to dryness, and dissolving the residue with 1ml methanol to obtain sample solution. Taking ginsenoside Rb1Reference substance, ginsenoside Re reference substance, and notoginsenoside R1Reference substance and ginsenoside Rg1As a control, a mixed solution containing 0.5mg of methanol per 1ml was prepared. Performing thin layer chromatography (general rule 0502) test, sucking 2 μ l of the above two solutions, respectively dropping on the same silica gel G thin layer plate, developing with chloroform-methanol-water (65:35:10) lower layer solution at 10 deg.C or below as developing agent, taking out, air drying, spraying 10% sulphuric acid ethanol solution, and heating at 105 deg.C until spots are clear. 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.
Example 6
The thin-layer identification method of the rhizoma alismatis comprises the following steps:
taking 6g of the product, grinding, adding 30ml of methanol, performing ultrasonic treatment for 30 minutes, filtering, and evaporating the filtrate to dryness. Dissolving the residue with 20ml water, transferring into separating funnel, extracting with ethyl acetate for 3 times (20 ml each time), mixing ethyl acetate solutions, evaporating to dryness, and dissolving the residue with 1ml methanol to obtain sample solution. Taking 2g of rhizoma alismatis as a reference medicinal material, adding 30ml of ethyl acetate, performing ultrasonic treatment for 30 minutes, filtering, evaporating filtrate to dryness, and dissolving residues by adding 1ml of methanol to obtain a reference medicinal material solution. Performing thin layer chromatography (general rule 0502) test, sucking 3 μ l of the above two solutions, respectively dropping on the same silica gel G thin layer plate, developing with cyclohexane-ethyl acetate (1:1) as developing agent, taking out, air drying, spraying with 5% silicotungstic acid ethanol solution, and heating at 105 deg.C until the spots are clearly developed. Spots of the same color appear on the chromatogram of the test solution at the positions corresponding to those on the chromatogram of the control solution.
Example 7
The notoginseng in the qi-tonifying and collateral-dredging granules is a valuable medicinal material with a large dosage, the main component of the qi-tonifying and collateral-dredging granules is saponins, the content determination method of the notoginseng and the ginseng in pharmacopeia is adopted for determination, and the components in a sample are enriched and purified by methods such as macroporous adsorption resin, solid phase extraction column and the like, but because the determination wavelength is 203nm, other medicinal components in the compound have interference, and the satisfactory separation effect cannot be achieved. Therefore, the characteristic saponin component with activity in the panax notoginseng medicinal material is finally selected to be detected by an evaporative light scattering detector, and the quality standard of the preparation is researched. To ensure the effectiveness of the preparation.
1.1 instruments, reagents
HPLC, Agilent 1200; evaporative light scattering detector: ELSD 2000 SE; acetonitrile: DikmaPure; and (4) self-preparing high-purity water. Ginsenoside Rb1(Ginsenoside Rb1The batch number is: 110704-201122), Ginsenoside Re (Ginsenoside Re, batch No.: 110754-1(Notoginsenoside R1The batch number is: 110745 and 200617), ginsenoside Rg1(Ginsenoside Rg1Lot 110703-201128) was provided by the china drug biologics assay.
1.2 chromatographic conditions
An Agilent 1200 high performance liquid chromatograph, a G1311A quaternary pump, a G1329A autosampler, a G1316A column oven, an ELSD-2000SE evaporation photodetector; a chromatographic column: MERCK RP-184.6 × 250mm (5 μm); temperature of the drift tube: 105 ℃; air flow rate: 2.4 ml/min; column temperature: 35 ℃ is carried out. Mobile phase: acetonitrile-water
TABLE 1 mobile phase chromatography elution ratio
1.3 methodological investigation
1.3.1 selection of extraction solvent: methanol is used as an extraction solvent according to pharmacopoeia and reference literature.
1.3.2 selection of extraction method: grinding the content of the product (batch No. 20120801), weighing about 5g, three parts, precisely weighing, placing in a conical flask, precisely adding 50ml of methanol solution, weighing, and ultrasonically treating for 45 min; heating and refluxing for 2 hours, preparing test solution I and II in sequence according to the text method, and the measurement results are shown in Table 1.
TABLE 2 selection of extraction methods
The results show that the total amount of the ginsenoside extracted by heating and refluxing is higher than that of the ginsenoside extracted by ultrasonic treatment, so the heating and refluxing extraction is adopted in the text.
1.3.3 selection of extraction time: grinding the content of the product (lot 20120801), weighing about 5g, three parts, precisely weighing, placing in a conical flask, precisely adding 50ml of methanol solution, weighing, and reflux-extracting for 1, 2, and 3 hours; test solutions were prepared according to the text method, and the results are shown in Table 2.
TABLE 3 different extraction times
The results show that the heating reflux extraction is complete after 2 hours, so the text adopts the heating reflux extraction for 2 hours.
1.3.4 blank test
Preparing blank solution according to the proportion of Chinese medicinal materials in the prescription, preparing group medicine without Notoginseng radix, preparing blank preparation according to the process, preparing and measuring according to the preparation method of test solution, and determining the blank solution and notoginsenoside R1Reference substance, ginsenoside Rg1Reference substance and ginsenoside Rb1No significant chromatographic peak was observed at the same retention time as the control, and the blank formulation was not interfering with the assay.
1.3.5 Linear relationship investigation
Collecting notoginsenoside R1Reference substance, ginsenoside Rg1Reference substance and ginsenoside Rb1Adding appropriate amount of reference substance, precisely weighing, and adding methanol to obtain a solution containing notoginsenoside R per 1ml11.28mg, ginsenoside Rg13.9mg of ginsenoside Rb13.4mg of the mixed solution, precisely sucking 10, 25, 50, 100, 125, 150 and 200 mul of the mixed solution respectively, placing the mixed solution into a 1ml measuring flask, adding methanol to dilute the mixed solution to a scale, shaking up the mixed solution, sucking 1Injecting 0 mu l of the mixture into a liquid chromatograph, continuously injecting the sample for 2 times, and measuring the peak area, wherein the result is shown in tables 4-6.
TABLE 4 notoginsenoside R1Measurement results
TABLE 5 ginsenoside Rg1Measurement results
TABLE 6 ginsenoside Rb1Measurement results
The result shows that the notoginsenoside R1Good linearity in the range of 0.32-2.56 μ g; ginsenoside Rg1Good linearity in the range of 0.39-5.85 μ g and ginsenoside Rb1The linearity is good within the range of 0.34 to 5.1 μ g. Regression equation of notoginsenoside R1Is Y-1.5887X-5.2358, r-0.9990; ginsenoside Rg1Is Y-1.5638X-4.7393, r-0.9996; ginsenoside Rb1Y is 1.5728X-4.5978 and r is 0.9994.
1.3.6 precision test
Preparing sample solution from the same batch of samples (lot number 20120801) according to the text method, precisely sucking the same sample solution, introducing sample 10 μ l under determined HPLC-ELSD condition, repeatedly introducing sample 5 times to obtain notoginsenoside R1Ginsenoside Rg1And ginsenoside Rb1The relative standard deviation of the natural logarithm of the peak area was < 2%, and the results are shown in Table 7.
TABLE 7 results of precision test
1.3.7 stability test
The same batch of samples (batch No. 20120801) was prepared into test solution according to the text method, and 5. mu.l of the same test solution was precisely absorbed and measured according to the method at 0, 2, 4, 6, 8, and 24 hours after preparation, indicating that the test solution was substantially stable within 24 hours. The results are shown in Table 8.
TABLE 8 stability test results
1.3.8 repeatability test
The same batch of samples (batch No. 20120801) was taken and tested as indicated in the text to give 6 test solutions and the results are shown in Table 9 under defined ELSD-HPLC conditions with a relative standard deviation of < 3%, indicating good reproducibility of the method.
TABLE 9 results of the repeatability tests
1.3.9 recovery test:
precisely weighing samples (lot number: 20120801, content of notoginsenoside R) with known content of same lot by sample loading and recovering method111.84mg/10g of ginsenoside Rg139.12mg/10g of ginsenoside Rb127.96mg/10g) about 2.5g, and precisely adding control mixed solution (notoginsenoside R)10.0868mg/ml, ginsenoside Rg10.24mg/ml ginsenoside Rb10.15604mg/ml)50ml, preparing according to the preparation method of the test solution and the chromatographic conditions, measuring, calculating the recovery rate according to the following formula, and obtaining the average recovery rate of notoginsenoside R199.79%; ginsenoside Rg1 is 98.34%; the content of ginsenoside Rb1 is 100.5%, the relative standard deviation is 3.0%, which shows that the method is reliable, and the results are shown in tables 10-12.
TABLE 10 notoginsenoside R1Recovery test
TABLE 11 ginsenoside Rg1Recovery test
TABLE 12 ginsenoside Rb1Recovery test
1.3.10 Total Saponin content in sample
Determination of notoginsenoside R according to text load method1Ginsenoside Rg1And ginsenoside Rb1The total amount of the (D) is calculated by an external standard two-point method logarithmic equation to obtain the final product. The results are shown in Table 13.
TABLE 13 results of sample measurement
1.3.11 sample content limit specification
The Chinese pharmacopoeia stipulates that: the Notoginseng radix contains ginsenoside Rg calculated according to dry product1(C42H72O14) Ginsenoside Rb1(C54H92O23) And notoginsenoside R1(C47H80O18) The total amount of (A) should not be less than 5.0%. When the materials are actually fed, the total amount of the three is much higher than the limit specified by pharmacopoeia, so that the ginsenoside Rg in the finished product1(C42H72O14) Ginsenoside Rb1(C54H92O23) And Notoginseng radix soapGlycoside R1(C47H80O18) The total amount is also high. In order to make the content limit of the prepared sample have more universal adaptive significance, the ginsenoside Rg is planned to be regulated according to pharmacopoeia1(C42H72O14) Ginsenoside Rb1(C54H92O23) And notoginsenoside R1(C47H80O18) The total amount of the ginsenoside Rg is set to be standard at the lowest limit, the transfer rate is calculated according to 100 percent, and each bag of ginsenoside Rg is converted1(C42H72O14) Ginsenoside Rb1(C54H92O23) And notoginsenoside R1(C47H80O18) The total amount of (A) should not be less than 50 mg/bag.
Example 8
2, measuring the content of formononetin:
2.1 instruments, reagents
LC-20AT liquid chromatograph; an ultraviolet detector; methanol: DikmaPure; and (4) self-preparing high-purity water. Formononetin (Formononetin, batch No.: 111703-200603) was provided by the China pharmaceutical biologicals assay.
2.2 methodological investigation
2.2.1 examination of extraction
The preparation method comprises precisely weighing the above (lot number: 20120801) granules, grinding, weighing about 1g, weighing 4 parts, adding 25mL methanol, weighing, and extracting by ultrasonic and refluxing methods, wherein each method comprises two parts in parallel. Extracting for 30min for 1 time, filtering, evaporating solvent, dissolving with methanol, and diluting to 10 ml.
The standard solution and the sample solution with the concentration of 2.1 mug/ml, 4.2 mug/ml are respectively and precisely absorbed with 10 mug each, and then injected into a high performance liquid chromatograph for content determination, and the result is shown in Table 14.
TABLE 14 examination results of various extraction methods
And (4) conclusion: compared with the reflux method, the reflux method has higher content of formononetin, so the reflux method is selected for extraction.
2.2.2 investigation of solvent dosage
Precisely weighing the product (batch number: 20120801) particles, grinding into fine powder, placing 1g of the powder into a 50ml conical flask with a plug, adding 30ml and 40ml of methanol respectively, refluxing for 30min, filtering, evaporating the solvent to dryness, adding a proper amount of methanol for dissolving, fixing the volume to a 10ml volumetric flask, and filtering with a 0.45 mu m filter membrane to obtain a sample solution.
Respectively and precisely sucking 10 μ l of each of the standard solution and the sample solution, and injecting into a high performance liquid chromatograph for content determination, wherein the results are shown in Table 15.
TABLE 15 examination of solvent dosages
Extracting by reflux method, and adding 25ml, 30ml and 40ml of methanol under certain time and frequency conditions, wherein the content of formononetin is small, so that 25ml of methanol is selected for extraction.
2.2.3 examination of extraction time
4 parts of granules are precisely weighed, 1g of each part is divided into two groups, and each group is divided into two parts for parallel experiments. Adding methanol 25ml, reflux extracting for 45min and 1 hr respectively, extracting for one time, filtering the extractive solution, evaporating to dryness under reduced pressure, adding appropriate amount of methanol to dissolve, and diluting to 10ml, filtering with 0.45 μm filter membrane to obtain sample solution.
Respectively and precisely sucking 10 μ l of each of the standard solution and the sample solution, and injecting into a high performance liquid chromatograph for content determination, wherein the results are shown in Table 16.
TABLE 16 examination of extraction times
From the results it can be seen that: extracting with 25ml under reflux for 30min, 45min, and 60min, wherein the content measured in 45min is close to that measured in 60min, and the content measured in 45min is higher than that measured in 30min, so the extraction time is 45 min.
2.2.4 examination of extraction times
6 parts of granules are precisely weighed, 1g of each part is divided into three groups, and two parts of each group are parallel. Adding 25ml of methanol into each part, performing reflux extraction for 45min for 1, 2 and 3 times, performing rotary evaporation under reduced pressure to remove the solvent, adding methanol to dissolve, metering to 10ml volumetric flask, and filtering with 0.45 μm filter membrane to obtain sample solution.
Respectively and precisely sucking 10 μ l of each of the standard solution and the sample solution, and injecting into a high performance liquid chromatograph for content determination, wherein the results are shown in Table 17.
TABLE 17 examination of the number of extractions
From the experimental results, the content of the methanol solution which is extracted twice and three times is close to the content of the methanol solution which is extracted once, and the number 6 value is lower, because a small part of the methanol solution is not completely transferred when the methanol solution is transferred from the eggplant-shaped bottle to the volumetric flask for constant volume. Finally, the reflux method is considered to be better for 2 times of extraction.
And (4) conclusion: reflux extracting with methanol 25ml for 45min for 2 times.
2.2.5 blank test
Preparing a blank preparation according to the traditional Chinese medicine proportion of the prescription, preparing a blank sample solution according to the selected method, and injecting the sample, wherein the blank sample solution does not have an obvious chromatographic peak at the same retention time as an formononetin reference substance, and the blank preparation has no interference to measurement.
2.2.6 Linear Range test
Weighing control formononetin 5.03mg, and diluting to 100ml with methanol to obtain control solution of 50.3 μ g/ml as stock solution. 1ml of the stock solution was taken and respectively stored in 2, 5, 10 and 25ml volumetric flasks to obtain the concentrations of 25.15, 10.06, 5.03 and 2.012. mu.g/ml.
0.5ml of stock solution is respectively taken and respectively prepared in 2, 10 and 25ml volumetric flasks to obtain the concentrations of 12.575, 2.515 and 1.006 mu g/ml. A total of 8 concentrations were obtained: 1.006, 2.012, 2.515, 5.03, 10.06, 12.575, 25.15, 50.3 μ g/ml. Samples were taken at 8 concentrations, 10. mu.l each, and the results are shown in Table 18.
TABLE 18 precision test results
The result shows that the formononetin is in the range of 1.006-50.3 mu g/ml, and the linearity is good. The regression equation is 74636X-4894, r 0.9998.
2.2.7 precision test
Precisely weighing 1g of the granule, preparing according to the preparation method of the test sample, repeatedly injecting sample for 6 times according to the formulated chromatographic conditions, 10 μ l each time, measuring the peak area, and the result is shown in Table 19.
TABLE 19 results of precision test
The result shows that the method has good precision.
2.2.8 repeatability test
Precisely weighing 6 parts of the same batch of granules, each part of the granules is 1g, preparing a test solution according to the same preparation method, injecting 10 mu l of each sample, measuring the peak area of 6 parts, and calculating the content of formononetin, which is shown in a table 20.
TABLE 20 results of the repeatability tests
The results show that the method has good repeatability.
2.2.9 stability test
Taking one part of the same batch of granules, preparing a test solution according to the method, precisely sucking 10 mu l of the test solution at 0, 2, 4, 6, 8, 16 and 24 hours respectively, and measuring the peak area, which is shown in Table 21.
TABLE 21 precision test results
The results show that the test solution is stable within 24 hours.
2.2.10 sample recovery test
6 parts (lot: 20120801) of the test sample of known content were precisely weighed, 0.5g of each part was added to a predetermined amount of the control solution at a ratio of 1:1, and the contents of the components were measured in order to determine the recovery rate of the sample application, the results are shown in Table 22.
TABLE 22 sample recovery test
2.2.11 sample determination
Taking 3 batches of samples to be tested, preparing the samples according to the preparation method of the samples, precisely sucking 10 mu l of sample solution to be tested respectively, injecting samples, measuring according to the conditions, and calculating the content in the 3 batches of samples, wherein the results are shown in a table 23.
TABLE 23 results of sample measurement
2.2.12 sample content limit specification
Calculating transfer rate according to 60% based on the content of formononetin in the sample, and calculating to obtain a bag containing formononetin (C)21H18O12) Calculated, not less than 0.25 mg.
Claims (4)
1. A detection method of a traditional Chinese medicine composition for treating qi deficiency and blood stasis syndrome is characterized in that the traditional Chinese medicine composition is prepared from the following raw medicinal materials of 500g of hedysarum polybotrys, 100g of pseudo-ginseng, 333g of rhizoma alismatis, 333g of angelica sinensis, 200g of ligusticum wallichii, 100g of cicada slough and 200g of radix curcumae and 100g of cassia twig; the method for detecting the traditional Chinese medicine composition is characterized by comprising a thin-layer identification method of hedysarum polybotrys, cassia twig, ligusticum wallichii, angelica sinensis, pseudo-ginseng and rhizoma alismatis and a content determination method of notoginsenoside R1 and formononetin;
the thin-layer identification method of the radix hedysari and the cassia twig comprises the following steps:
(1) the method for identifying the radix hedysari comprises the following steps: grinding the Chinese medicinal composition, adding methanol, filtering, evaporating filtrate to dryness, dissolving residue in water, extracting with diethyl ether, evaporating to dryness, and dissolving residue in methanol to obtain sample solution; adding methanol into formononetin reference substance to obtain reference substance solution; and (3) performing thin-layer chromatography test, namely respectively dropping the two solutions on the same silica gel thin-layer plate according to the ratio of petroleum ether to ethyl acetate of 0.5-2: 0.5-2 of developing agent, presaturating with developing agent, developing, taking out, drying, and inspecting under 254nm ultraviolet lamp to obtain spots with same color in the chromatogram of the test sample and the corresponding positions of the chromatogram of the reference sample;
(2) the method for identifying the cassia twig comprises the following steps: taking a cassia twig reference medicinal material, preparing a reference medicinal material solution according to the preparation method of the test solution under the item of the identification (1), and adding methanol into a cinnamic acid reference substance to prepare a solution serving as a reference substance solution; and (2) performing thin-layer chromatography test, namely absorbing the sample solution and the two solutions under the identification item (1), respectively dropping the sample solution and the two solutions on the same silica gel thin-layer plate, and performing thin-layer chromatography test on the sample solution by using petroleum ether-n-hexane-ethyl formate-formic acid 1.5-2.5: 5-7: 2-4: 0.1-0.3 of developing agent, developing, taking out, drying, and inspecting under an ultraviolet lamp at 254nm, wherein spots with the same color appear in the chromatogram of the test solution at the positions corresponding to the chromatograms of the reference medicinal material and the reference solution;
the content determination method of the notoginsenoside R1 comprises the following steps:
the method comprises the following steps of: acetonitrile is used as a mobile phase A, water is used as a mobile phase B, and gradient elution is carried out according to the following elution proportion: time, 0-20 minutes, mobile phase a: 21 → 50%, mobile phase B: 79 → 50%;
preparing a control solution: precisely weighing appropriate amount of notoginsenoside R1, ginsenoside Rg1 and ginsenoside Rb1, adding methanol to obtain mixed solution containing 10.1-0.3 mg of notoginsenoside R, 10.4-0.6 mg of ginsenoside Rg and 10.3-0.5 mg of ginsenoside Rb per 1ml,
preparing a test sample solution: mixing the Chinese medicinal composition, grinding, accurately weighing, adding methanol, weighing, refluxing in water bath for 1-3 hr, cooling, weighing, adding methanol to balance weight, shaking, filtering, collecting filtrate,
fourth, measurement method: precisely sucking the reference solution and the sample solution, respectively, injecting into a liquid chromatograph, and measuring.
2. The method for detecting the traditional Chinese medicine composition as claimed in claim 1, wherein the thin layer identification method of ligusticum wallichii, angelica sinensis, panax notoginseng and rhizoma alismatis in the detection method is any one or combination of more of the following items (1) to (4):
(1) the identification method of the ligusticum wallichii comprises the following steps: grinding the Chinese medicinal composition, adding methanol, filtering, evaporating filtrate to dryness, dissolving residue in water, extracting with diethyl ether, evaporating to dryness, and dissolving residue in methanol to obtain sample solution; taking a ligusticum wallichii reference medicinal material, grinding, adding methanol, filtering, evaporating filtrate to dryness, dissolving residue by adding water, adding diethyl ether for extraction, evaporating to dryness, dissolving residue by adding methanol to prepare a reference medicinal material solution, performing a thin-layer chromatography test, sucking and identifying a sample solution and the reference medicinal material solution, respectively dropping the sample solution and the reference medicinal material solution on the same silica gel thin-layer plate, and adding n-hexane-ethyl acetate 0.8-1.2: 0.8-1.2 of developing agent, developing, taking out, drying, placing under an ultraviolet lamp at 365nm for inspection, and displaying fluorescent spots with the same color in the chromatogram of the test solution at the position corresponding to the chromatogram of the reference solution;
(2) the identification method of the angelica comprises the following steps: taking ferulic acid reference substance, adding methanol to prepare solution, taking the solution as reference substance solution, performing thin-layer chromatography test, sucking the sample solution and the reference substance solution under the item (1) for identification, respectively dropping the sample solution and the reference substance solution on the same silica gel thin-layer plate, and adding 3.5-4.5 parts of cyclohexane-trichloromethane-ethyl acetate-formic acid: 0.8-1.2: 3.5-4.5: 0.05-0.15 of developing agent, developing, taking out, airing, and placing under an ultraviolet lamp at 254nm for inspection, wherein spots with the same color appear in the chromatogram of the test solution at the positions corresponding to the chromatograms of the reference solution;
(3) the identification method of the pseudo-ginseng comprises the following steps: taking the traditional Chinese medicine composition, grinding, adding methanol, filtering, evaporating filtrate to dryness, dissolving residues in water, adding water saturated n-butanol for extraction, washing with ammonia test solution, evaporating n-butanol solution to dryness, dissolving residues in methanol to obtain a test solution, taking a ginsenoside Rb1 reference substance, a ginsenoside Re reference substance, a notoginsenoside R1 reference substance and a ginsenoside Rg1 reference substance, adding methanol to prepare a mixed solution containing 0.5mg of ginsenoside Rb1 in each 1ml, taking the mixed solution as a reference solution, performing a thin-layer chromatography test, sucking the two solutions, respectively dropping the two solutions on a same silica gel thin-layer plate, and dissolving the mixed solution in chloroform-methanol-water by 60-70: 32-37: 8-12, taking out, airing, spraying a 10% sulfuric acid ethanol solution, heating at 105 ℃ until spots are clear, wherein spots with the same color appear in the chromatogram of the test sample at the position corresponding to the chromatogram of the reference sample;
(4) the identification method of the rhizoma alismatis comprises the following steps: grinding the Chinese medicinal composition, adding methanol, ultrasonic treating, filtering, evaporating filtrate to dryness, dissolving residue in water, extracting with ethyl acetate, evaporating to dryness, and dissolving residue in methanol to obtain sample solution; adding ethyl acetate into rhizoma alismatis reference medicinal material, performing ultrasonic treatment, filtering, evaporating filtrate to dryness, dissolving residue by adding methanol to obtain reference medicinal material solution, performing thin-layer chromatography test, sucking the two solutions, respectively dropping the two solutions on the same silica gel thin-layer plate, and adding cyclohexane-ethyl acetate 0.8-1.2: 0.8-1.2 of developing agent, developing, taking out, drying, spraying 5% silicotungstic acid ethanol solution, heating at 105 ℃ until the spots are clear, and showing spots with the same color in the chromatogram of the test solution at the position corresponding to the chromatogram of the reference medicinal material.
3. The method for detecting the traditional Chinese medicine composition of claim 1, wherein the method for detecting the content of formononetin in the traditional Chinese medicine composition comprises the following steps:
the method comprises the following steps of: 55-60% of methanol-water: 40-45 is a mobile phase; the detection wavelength is 250nm, and the column temperature is 25-35 ℃;
preparing a control solution: taking a proper amount of formononetin reference substances, precisely weighing, and adding methanol to prepare a solution containing 6-10 mu g of formononetin per 1ml to obtain the formononetin reference substances;
preparing a test sample solution: mixing the Chinese medicinal composition, grinding, accurately weighing, placing in a conical flask, adding methanol, refluxing in water bath, filtering, placing the residue and filter paper in the conical flask, adding methanol, refluxing, mixing the filtrates, washing the filter paper with methanol, concentrating under reduced pressure, metering the volume of the concentrated solution to a measuring flask, shaking, filtering, and collecting the filtrate.
4. The method for detecting a traditional Chinese medicine composition according to claim 3, wherein the chromatographic conditions in the step are as follows: with methanol-water 58:42 is a mobile phase; the detection wavelength is 250nm, and the column temperature is 30 ℃.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810328487.4A CN110412198B (en) | 2018-04-28 | 2018-04-28 | Detection method of traditional Chinese medicine composition for treating qi deficiency and blood stasis syndrome |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810328487.4A CN110412198B (en) | 2018-04-28 | 2018-04-28 | Detection method of traditional Chinese medicine composition for treating qi deficiency and blood stasis syndrome |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110412198A CN110412198A (en) | 2019-11-05 |
| CN110412198B true CN110412198B (en) | 2021-08-31 |
Family
ID=68345611
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810328487.4A Active CN110412198B (en) | 2018-04-28 | 2018-04-28 | Detection method of traditional Chinese medicine composition for treating qi deficiency and blood stasis syndrome |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110412198B (en) |
Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1511577A (en) * | 2002-12-30 | 2004-07-14 | 红 张 | Medicine for treating primary and secondary headache |
| CN1806846A (en) * | 2005-01-18 | 2006-07-26 | 北京凯瑞创新医药科技有限公司 | Chinese medicinal composition, its preparation process and quality control method |
| CN101690752A (en) * | 2008-06-06 | 2010-04-07 | 广西灵峰药业有限公司 | Quality control method of medicine composition |
| CN101829232A (en) * | 2010-05-27 | 2010-09-15 | 王喜军 | Traditional Chinese medicine composition for treating thromboangiitis obliterans and preparation method thereof |
| CN102133377A (en) * | 2011-03-22 | 2011-07-27 | 唐秋海 | Quality detection method of capsule for reducing blood fat and activating collaterals |
| CN104922587A (en) * | 2015-02-12 | 2015-09-23 | 贵阳中医学院第二附属医院 | Preparation method of swelling-removing and pain-relieving paste and quality-detecting method of swelling-removing and pain-relieving paste |
| CN105259295A (en) * | 2015-11-17 | 2016-01-20 | 山东沃华医药科技股份有限公司 | Quality detection method for ginseng, cassia twig and poria cocos oral solution |
| CN105477166A (en) * | 2015-12-29 | 2016-04-13 | 广东一方制药有限公司 | Preparation method of pubescent angelica and mistletoe decoction formula granules and quality control method thereof |
| CN105535237A (en) * | 2015-12-29 | 2016-05-04 | 广东一方制药有限公司 | Preparation method and quality control method for peach-kernel-safflower decoction-of-four-ingredients formula granules |
| CN106290697A (en) * | 2016-08-29 | 2017-01-04 | 贵州信邦制药股份有限公司 | The discrimination method of Rhizoma Chuanxiong in SHIQUAN DABU JIU |
| CN107753724A (en) * | 2016-08-17 | 2018-03-06 | 江苏康缘药业股份有限公司 | A kind of preparation method and detection method of the tablet for treating urarthritis |
-
2018
- 2018-04-28 CN CN201810328487.4A patent/CN110412198B/en active Active
Patent Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1511577A (en) * | 2002-12-30 | 2004-07-14 | 红 张 | Medicine for treating primary and secondary headache |
| CN1806846A (en) * | 2005-01-18 | 2006-07-26 | 北京凯瑞创新医药科技有限公司 | Chinese medicinal composition, its preparation process and quality control method |
| CN101690752A (en) * | 2008-06-06 | 2010-04-07 | 广西灵峰药业有限公司 | Quality control method of medicine composition |
| CN101829232A (en) * | 2010-05-27 | 2010-09-15 | 王喜军 | Traditional Chinese medicine composition for treating thromboangiitis obliterans and preparation method thereof |
| CN102133377A (en) * | 2011-03-22 | 2011-07-27 | 唐秋海 | Quality detection method of capsule for reducing blood fat and activating collaterals |
| CN104922587A (en) * | 2015-02-12 | 2015-09-23 | 贵阳中医学院第二附属医院 | Preparation method of swelling-removing and pain-relieving paste and quality-detecting method of swelling-removing and pain-relieving paste |
| CN105259295A (en) * | 2015-11-17 | 2016-01-20 | 山东沃华医药科技股份有限公司 | Quality detection method for ginseng, cassia twig and poria cocos oral solution |
| CN105477166A (en) * | 2015-12-29 | 2016-04-13 | 广东一方制药有限公司 | Preparation method of pubescent angelica and mistletoe decoction formula granules and quality control method thereof |
| CN105535237A (en) * | 2015-12-29 | 2016-05-04 | 广东一方制药有限公司 | Preparation method and quality control method for peach-kernel-safflower decoction-of-four-ingredients formula granules |
| CN107753724A (en) * | 2016-08-17 | 2018-03-06 | 江苏康缘药业股份有限公司 | A kind of preparation method and detection method of the tablet for treating urarthritis |
| CN106290697A (en) * | 2016-08-29 | 2017-01-04 | 贵州信邦制药股份有限公司 | The discrimination method of Rhizoma Chuanxiong in SHIQUAN DABU JIU |
Non-Patent Citations (6)
| Title |
|---|
| HPLC 法测定活血止痛胶囊中三七皂苷R1、人参皂苷Rg1、人参皂苷Rb1及阿魏酸;韩杰 等;《中成药》;20110731;第33卷(第7期);1186-1189 * |
| HPLC测定新疆红芪中芒柄花素的含量;海力茜 等;《华西药学杂志》;20051231;第20卷(第5期);433-434 * |
| 心可舒丸中三七的薄层色谱鉴别和6种物质的含量测定;赵振霞 等;《华西药学杂志》;20161231;第31卷(第6期);656-659 * |
| 泽泻薄层色谱鉴别研究;巩丽萍 等;《中国中药杂志》;20061031(第19期);1628-1629 * |
| 赵振霞 等.心可舒丸中三七的薄层色谱鉴别和6种物质的含量测定.《华西药学杂志》.2016,第31卷(第6期), * |
| 龙胆泻肝丸(大蜜丸)薄层色谱鉴别方法的建立;赵顺 等;《中国药师》;20151231;第18卷(第1期);151-153 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110412198A (en) | 2019-11-05 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111044624B (en) | Quality detection method of Chinese medicinal preparation | |
| CN101850070B (en) | Detection method for Chinese medicament Tangcao tablets | |
| CN101167788B (en) | Quality control method of traditional Chinese medicine 'zhenqi fuzheng' containing glossy privet fruit and radix astragali for strengthening the body resistance for aeipathia deficiency damage and qi | |
| CN107991425B (en) | Detection method of traditional Chinese medicine composition for treating traumatic injury | |
| CN102145041B (en) | Pharmaceutical composition with function of treating polycystic ovary syndrome | |
| CN106822203B (en) | Radix angelicae pubescentis granules and preparation method and quality control method thereof | |
| CN113063885B (en) | Composition for preparing Baoyuan decoction, baoyuan decoction product and fingerprint spectrum measuring and quality detecting method thereof | |
| CN101028388B (en) | Quality inspection of Chinese-medicinal preparation for treating shortsighness and asthenopia | |
| CN102426205B (en) | Detection method of thrombus sumalin capsules | |
| CN105628851B (en) | A kind of detection method of Chinese medicine preparation | |
| CN100388940C (en) | Quality control method of Chinese medicinal preparation for treating child hyperpyrexia | |
| CN102091297A (en) | Quality control method for liver health care medicine | |
| CN110412198B (en) | Detection method of traditional Chinese medicine composition for treating qi deficiency and blood stasis syndrome | |
| CN108414667B (en) | Method for detecting quality standard of Shengui Yixin granules | |
| CN101700306A (en) | Quality control method of Rupixiao preparation | |
| CN113759011B (en) | Method for establishing characteristic spectrum of starwort root and preparation thereof | |
| CN112526045B (en) | Method for simultaneously detecting or identifying effective components in heart-soothing and lipid-lowering tablets | |
| CN107966505A (en) | The method for measuring Multiple components in Fufang Danshen Pian at the same time | |
| CN108614066A (en) | A kind of Traditional Chinese medicine composition detection method for treating coronary heart disease | |
| CN102809626A (en) | Quality control method of Qianshan Huoxue plaster | |
| CN106018661A (en) | Quality control method of raw astragalus membranaceus wasting-thirst capsules | |
| CN115166066B (en) | Quality evaluation method of Qizhu oral liquid for improving white blood cells | |
| CN114384165B (en) | Quality control method of osmanthus dragon ointment | |
| CN117110449A (en) | HPLC method for determining stilbene glucoside content in TONGBIHUOLUO pill | |
| CN112843110A (en) | Preparation method and quality standard of pseudo-ginseng powder formula granules |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |