CN107153098B - Method for measuring fingerprint spectrum of traditional Chinese medicine composition - Google Patents
Method for measuring fingerprint spectrum of traditional Chinese medicine composition Download PDFInfo
- Publication number
- CN107153098B CN107153098B CN201610119793.8A CN201610119793A CN107153098B CN 107153098 B CN107153098 B CN 107153098B CN 201610119793 A CN201610119793 A CN 201610119793A CN 107153098 B CN107153098 B CN 107153098B
- Authority
- CN
- China
- Prior art keywords
- parts
- acid
- mug
- traditional chinese
- chinese medicine
- 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
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/027—Liquid 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)
- Cosmetics (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention claims a high performance liquid chromatography fingerprint spectrum measuring method of a traditional Chinese medicine composition, and the traditional Chinese medicine composition is composed of the following medicinal materials: the fingerprint spectrum measuring method comprises the steps of calibrating 15 main chromatographic peaks by using forsythia, honeysuckle, isatis root, bitter apricot seed, menthol, houttuynia, rheum officinale, patchouli, male fern rhizome, rhodiola rosea, ephedra, liquorice and gypsum, and identifying 9 of the main chromatographic peaks as neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, iso-forsythiaside A, forsythoside A, phillyrin, quercetin, 4, 5-di-O-caffeoylquinic acid and glycyrrhizic acid.
Description
Technical Field
The invention relates to a method for measuring a fingerprint spectrum of a traditional Chinese medicine composition.
Background
The traditional Chinese medicine fingerprint refers to a chromatogram or a spectrogram which can mark chemical characteristics of certain traditional Chinese medicinal materials or traditional Chinese medicine preparations by adopting a certain analysis means after the traditional Chinese medicinal materials or the traditional Chinese medicine preparations are properly processed. The traditional Chinese medicine fingerprint spectrum is a comprehensive and quantifiable identification means, is established on the basis of the systematic research of the chemical components of the traditional Chinese medicine, and is mainly used for evaluating the authenticity, the excellence and the stability of the quality of the semi-finished products of the traditional Chinese medicine and the traditional Chinese medicine preparation. "integrity" and "fuzziness" are its distinguishing features. The fingerprint is taken as the quality control method of the traditional Chinese medicine (natural medicine) extract and the preparation thereof, which is the current international consensus, and various fingerprint control technical systems conforming to the characteristics of the traditional Chinese medicine (natural medicine) are researched and established. The Food and Drug Administration (FDA) allows the provision of chromatographic fingerprints in herbal health product declaration documentation; the World Health Organization (WHO) also stipulates in the 1996 herbal evaluation guidelines that if the active ingredients of herbs are not defined, a chromatographic fingerprint can be provided to demonstrate consistency in product quality; the European Community is also known in the herbal quality guide, and it is not sufficient to examine the stability of the quality by measuring a certain active ingredient alone, because herbs and their preparations are active substances as a whole. Chromatographic fingerprints, especially sharp fingerprints of thin layer chromatography, are useful. The application of foreign fingerprint aims at solving the problems of quality detection of plant medicines with complex components and uncertain effective components and quality difference among product batches.
High Performance Liquid Chromatography (HPLC) fingerprint has high separation degree, and can separate complex chemical components to form different peaks to form a chromatogram, and the heights and peak areas of the chromatogram peaks respectively represent various chemical components and the contents thereof. Therefore, the traditional Chinese medicine fingerprint is further developed than the DNA fingerprint: not only can the characteristic embodiment (the number and relative position of various chemical components-retention time) be used for qualitative identification, but also the concept of quantity is embodied. The height and area of the peak represent the content of a certain chemical component, and the ratio of the peak height (or peak area) of each peak represents the relative content of various chemical components; the introduction of the concept of quantity, the combination of qualitative and quantitative endows the fingerprint of the traditional Chinese medicine with greater efficacy; the traditional Chinese medicine fingerprint can not only identify the uniqueness of an individual and a certain species, but also hook the quantity characteristics of the individual and the species with other systems. Therefore, the traditional Chinese medicine fingerprint is not only a traditional Chinese medicine quality control mode and technology, but also can be developed into a research system and a research mode for developing traditional Chinese medicine theories (complex systems) and new drugs by adopting various fingerprints. The fingerprint spectrum should have fingerprint properties, namely: (1) the specificity is strong. The finger print prepared should be unique to the traditional Chinese medicine, can be distinguished from other traditional Chinese medicines, and reflects chemical information with high selectivity; (2) the stability is good. The fingerprint spectrum of the traditional Chinese medicine is the commonness induced from a plurality of batches of the traditional Chinese medicine, and the common peak or the characteristic peak in the spectrum is relatively stable; (3) the reproducibility is good. The fingerprint map is prepared to reproduce fingerprint characteristics (such as the number, the size, the position and the like of common peaks) under specified conditions, and the error of the fingerprint map is within an allowable range. Only then, the prepared fingerprint has practical value, and the quality of the medicine can be effectively controlled. The UPLC fingerprint method has the characteristics of sensitivity, rapidness, simplicity, convenience, accuracy and the like, and the fingerprint of the pharmaceutical preparation is determined by optimizing chromatographic conditions, so that the quality of the pharmaceutical can be well controlled.
The traditional Chinese medicine fingerprint spectrum can be used in various stages of research and production processes of traditional Chinese medicine preparations, is classified according to application objects, and can be divided into traditional Chinese medicine (raw medicinal materials) fingerprint spectrums, traditional Chinese medicine raw material medicine (including decoction pieces and compatible granules) fingerprint spectrums and traditional Chinese medicine preparation fingerprint spectrums. If the product is more finely divided, the fingerprint of the intermediate product in the process production process can also be included. It is worth noting that the establishment of the chemical fingerprint of Chinese medicinal materials must take into account the influence of many factors. The quality control is caused by the factors of the mixture of Chinese medicinal varieties, the history formation of homonymous foreign matters and heteronymous homonymous substances, the production place environment of the same species, the large fluctuation of components contained in different effective parts and the like. Therefore, for the fingerprint of a certain traditional Chinese medicinal material, a large amount of data needs to be accumulated, and a reasonable similarity standard can be formulated. The traditional Chinese medicine and the preparation thereof are all multi-component complex systems, so that the quality of the traditional Chinese medicine and the preparation thereof is evaluated by adopting a detection method which is adaptive to the traditional Chinese medicine and can provide rich identification information, but the existing methods such as microscopic identification, physicochemical identification, content measurement and the like are not enough to solve the problem, the establishment of the traditional Chinese medicine fingerprint spectrum can more comprehensively reflect the types and the quantities of chemical components contained in the traditional Chinese medicine and the preparation thereof, and further the overall description and evaluation of the quality of the medicine are carried out. This also corresponds to the holistic theory of traditional Chinese medicine. On the basis, if the research on the spectrum effect is further carried out, the quality of the traditional Chinese medicine and the drug effect thereof can be really combined, which is helpful for clarifying the action mechanism of the traditional Chinese medicine. In a word, the research and establishment of the traditional Chinese medicine fingerprint spectrum have important significance for improving the quality of the traditional Chinese medicine and promoting the modernization of the traditional Chinese medicine.
The patent application number is 201210450660.0, and the invention name is: a process for preparing antiviral Chinese-medicinal composition includes such steps as ultrasonic reflux extracting forsythia fruit, rhubarb, ephedra herb, houttuynia, calibrating 27 common peaks, and identifying 4 common peaks including forsythoside A, forsythin, pinoresinol-4-O-glc and rhein. The invention claims a high performance liquid chromatography fingerprint spectrum measuring method of a traditional Chinese medicine composition, and the traditional Chinese medicine composition is composed of the following medicinal materials: the fingerprint spectrum measuring method comprises the steps of calibrating 15 main chromatographic peaks by using forsythia, honeysuckle, isatis root, bitter apricot seed, menthol, houttuynia, rhubarb, patchouli, male fern rhizome, rhodiola rosea, ephedra, liquorice and gypsum, and identifying 9 of the 15 main chromatographic peaks as neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, iso-forsythiaside A, phillyrin, quercetin, 4, 5-di-O-caffeoylquinic acid, phillyrin and glycyrrhizic acid, wherein the technical content is not disclosed in the prior art.
Disclosure of Invention
The invention provides a fingerprint spectrum measuring method of a traditional Chinese medicine composition.
The traditional Chinese medicine composition is prepared from the following raw material medicines in parts by weight: 200-300 parts of fructus forsythiae, 60-100 parts of ephedra, 40-60 parts of rheum officinale, 200-300 parts of houttuynia cordata, 200-300 parts of honeysuckle, 200-300 parts of radix isatidis, 60-100 parts of pogostemon cablin, 200-300 parts of rhizoma dryopteris crassirhizomae, 60-100 parts of rhodiola rosea, 5-9 parts of menthol, 60-100 parts of bitter almond, 60-100 parts of liquorice and 200-300 parts of gypsum, and the fingerprint spectrum determination method is as follows:
preparing a test solution: precisely weighing 0.1-0.5 g of the traditional Chinese medicine composition, placing the traditional Chinese medicine composition in a conical flask with a plug, precisely adding 10-50mL of 40-100% methanol, weighing, ultrasonically treating for 20-40 minutes, cooling, weighing again, complementing the loss weight with 40-100% methanol, shaking up, filtering, and taking the subsequent filtrate to obtain the traditional Chinese medicine composition;
preparation of control solutions: respectively weighing appropriate amounts of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, iso-forsythiaside A, forsythiaside, 4, 5-di-O-caffeoylquinic acid, quercetin and ammonium glycyrrhizinate as control substances, and preparing a mixed control solution containing 18.4 mug/ml neochlorogenic acid, 27.2 mug/ml chlorogenic acid, 21.2 mug/ml cryptochlorogenic acid, 27.2 mug/ml iso-forsythiaside A, 21.6 mug/ml forsythiaside A, 16.0 mug/ml forsythiaside, 14.4 mug/ml 4, 5-di-O-caffeoylquinic acid, 9.0 mug/ml quercetin and 24.4 mug/ml ammonium glycyrrhizinate respectively to obtain the control substance solution;
preparation of reference solutions: taking a proper amount of forsythoside A reference substance, precisely weighing, and adding 50% methanol to prepare a solution containing 101.2 mug per 1ml to obtain the final product.
Chromatographic conditions are as follows: the mobile phase was methanol-acetonitrile-0.1% phosphoric acid, gradient elution, elution ratios such as the following table:
the chromatographic column is C18A chromatographic column; the detection wavelength is 239nm, and the flow rate is 0.1-0.5 ml/min;
the determination method comprises the following steps: precisely sucking 1-10 μ L of each of the reference solution and the sample solution, injecting into a ultra-high liquid chromatograph, measuring, and recording chromatogram.
The traditional Chinese medicine composition disclosed by the invention comprises the following raw material medicines in parts by weight: 200 parts of fructus forsythiae, 300 parts of honeysuckle, 200 parts of isatis root, 40 parts of rhubarb, 60 parts of cablin potchouli herb, 300 parts of male fern rhizome, 100 parts of rhodiola rosea, 9 parts of menthol, 60 parts of ephedra, 100 parts of bitter apricot seed, 200 parts of cordate houttuynia, 100 parts of liquorice and 200 parts of gypsum.
The traditional Chinese medicine composition disclosed by the invention comprises the following raw material medicines in parts by weight: 300 parts of fructus forsythiae, 200 parts of honeysuckle, 300 parts of isatis root, 60 parts of rheum officinale, 100 parts of pogostemon cablin, 200 parts of male fern rhizome, 60 parts of rhodiola rosea, 5 parts of menthol, 100 parts of ephedra, 60 parts of bitter apricot seed, 300 parts of houttuynia cordata, 60 parts of liquorice and 300 parts of gypsum.
The traditional Chinese medicine composition disclosed by the invention comprises the following raw material medicines in parts by weight: fructus forsythiae 278, honeysuckle 294, isatis root 285, rhubarb 55, patchouli 95, male fern 290, rhodiola rosea 87, menthol 8.5, ephedra 88, bitter apricot kernel 80, cordate houttuynia 284, liquorice 95 and gypsum 277.
The traditional Chinese medicine composition disclosed by the invention comprises the following raw material medicines in parts by weight: fructus forsythiae 255, honeysuckle 255, isatis root 255, rhubarb 51, patchouli 85, male fern 255, rhodiola rosea 85, menthol 7.5, ephedra 85, bitter apricot seed 85, cordate houttuynia 255, liquorice 85 and gypsum 255.
The preparation method of the traditional Chinese medicine composition preparation comprises the following steps:
(1) weighing the traditional Chinese medicinal materials according to the weight proportion of the raw materials, cleaning, and cutting off the traditional Chinese medicinal materials as appropriate;
(2) crushing herba Agastaches, extracting volatile oil with 10 times of water for 8 hr, and collecting volatile oil; filtering the extractive solution, discarding residue, and collecting filtrate;
(3) extracting fructus forsythiae, herba Ephedrae, herba Houttuyniae, and radix et rhizoma Rhei with 12 times of 70% ethanol for 3 times, each for 2.5 hr, mixing extractive solutions, filtering, and recovering ethanol to obtain filtrate;
(4) adding 12 times of water into honeysuckle, gypsum, isatis root, male fern rhizome, liquorice and rhodiola rosea, decocting until boiling, adding bitter apricot seed, decocting for 2 times, each time for 1 hour, combining extracting solutions, filtering, combining obtained filtrate with filtrate obtained after oil extraction of patchouli in the step (2), concentrating into clear paste with the relative density of 1.10-1.15 measured at the temperature of 60 ℃, adding ethanol, adjusting the alcohol concentration to 70%, refrigerating, standing, filtering, and recovering the ethanol until no alcohol smell exists, thus obtaining the clear paste for later use;
(5) mixing the fluid extract obtained in step (4) with the ethanol extract obtained in step (3), concentrating to obtain fluid extract with relative density of 1.15-1.20 at 60 deg.C, and drying to obtain dry extract powder;
(6) adding a proper amount of pharmaceutically acceptable auxiliary materials into the dry paste powder obtained in the step (5) for granulation;
(7) and (3) adding menthol and the volatile oil obtained in the step (2) into ethanol for dissolving, spraying the particles obtained in the step (6), sealing, uniformly mixing, and tabletting or encapsulating or bagging.
The fingerprint spectrum measuring method of the invention is preferably as follows:
preparing a test solution: taking 0.25g of the traditional Chinese medicine composition, precisely weighing, placing in a conical flask with a plug, precisely adding 25mL of 50% methanol, weighing, carrying out ultrasonic treatment (power 500W and frequency 40kHz) for 30 minutes, cooling, weighing again, supplementing the lost weight with 50% methanol, shaking up, filtering, and taking the subsequent filtrate to obtain the traditional Chinese medicine composition;
preparation of control solutions: respectively weighing appropriate amounts of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, iso-forsythiaside A, forsythiaside, 4, 5-di-O-caffeoylquinic acid, quercetin and ammonium glycyrrhizinate as control substances, and preparing a mixed control solution containing 18.4 mug/ml neochlorogenic acid, 27.2 mug/ml chlorogenic acid, 21.2 mug/ml cryptochlorogenic acid, 27.2 mug/ml iso-forsythiaside A, 21.6 mug/ml forsythiaside A, 16.0 mug/ml forsythiaside, 14.4 mug/ml 4, 5-di-O-caffeoylquinic acid, 9.0 mug/ml quercetin and 24.4 mug/ml ammonium glycyrrhizinate respectively to obtain the control substance solution;
preparation of reference solutions: taking a proper amount of forsythoside A reference substance, precisely weighing, and adding 50% methanol to prepare a solution containing 101.2 mug per 1ml to obtain the final product.
Chromatographic conditions are as follows: the mobile phase was methanol-acetonitrile-0.1% phosphoric acid, gradient elution, elution ratios such as the following table:
the chromatographic column is a Waters Acquity UPLC HSS T3 chromatographic column, the column length is 100mm, the inner diameter is 2.1mm, and the particle size is 1.8 mu m; the detection wavelength is 239nm, and the flow rate is 0.3 ml/min;
the determination method comprises the following steps: precisely sucking 2 mu L of each of the reference solution and the sample solution, injecting into an ultra-high liquid chromatograph, measuring, and recording a chromatogram.
The fingerprint spectrum measuring method of the invention is also preferably as follows:
preparing a test solution: taking 0.1 g of the traditional Chinese medicine composition, precisely weighing, placing in a conical flask with a plug, precisely adding 50mL of methanol, weighing, ultrasonically treating for 40 minutes, cooling, weighing again, supplementing the weight loss with methanol, shaking up, filtering, and taking the subsequent filtrate to obtain the traditional Chinese medicine composition;
preparation of control solutions: respectively weighing appropriate amounts of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, iso-forsythiaside A, forsythiaside, 4, 5-di-O-caffeoylquinic acid, quercetin and ammonium glycyrrhizinate as control substances, and preparing a mixed control solution containing 18.4 mug/ml neochlorogenic acid, 27.2 mug/ml chlorogenic acid, 21.2 mug/ml cryptochlorogenic acid, 27.2 mug/ml iso-forsythiaside A, 21.6 mug/ml forsythiaside A, 16.0 mug/ml forsythiaside, 14.4 mug/ml 4, 5-di-O-caffeoylquinic acid, 9.0 mug/ml quercetin and 24.4 mug/ml ammonium glycyrrhizinate respectively to obtain the control substance solution;
preparation of reference solutions: taking a proper amount of forsythoside A reference substance, precisely weighing, and adding 50% methanol to prepare a solution containing 101.2 mug per 1ml to obtain the final product.
Chromatographic conditions are as follows: the mobile phase was methanol-acetonitrile-0.1% phosphoric acid, gradient elution, elution ratios such as the following table:
the chromatographic column is a Waters Acquity UPLC HSS T3 chromatographic column; the detection wavelength is 239nm, and the flow rate is 0.1 ml/min;
the determination method comprises the following steps: precisely sucking 1 mu L of each of the reference solution and the sample solution, injecting into an ultra-high liquid chromatograph, measuring, and recording a chromatogram.
The fingerprint spectrum measuring method of the invention is also preferably as follows:
preparing a test solution: taking 0.5 g of the traditional Chinese medicine composition, precisely weighing, placing in a conical flask with a plug, precisely adding 10mL of 40% methanol, weighing, ultrasonically treating for 20 minutes, cooling, weighing again, supplementing the lost weight with 40% methanol, shaking uniformly, filtering, and taking the subsequent filtrate to obtain the traditional Chinese medicine composition;
preparation of control solutions: respectively weighing appropriate amounts of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, iso-forsythiaside A, forsythiaside, 4, 5-di-O-caffeoylquinic acid, quercetin and ammonium glycyrrhizinate as control substances, and preparing a mixed control solution containing 18.4 mug/ml neochlorogenic acid, 27.2 mug/ml chlorogenic acid, 21.2 mug/ml cryptochlorogenic acid, 27.2 mug/ml iso-forsythiaside A, 21.6 mug/ml forsythiaside A, 16.0 mug/ml forsythiaside, 14.4 mug/ml 4, 5-di-O-caffeoylquinic acid, 9.0 mug/ml quercetin and 24.4 mug/ml ammonium glycyrrhizinate respectively to obtain the control substance solution;
preparation of reference solutions: taking a proper amount of forsythoside A reference substance, precisely weighing, and adding 50% methanol to prepare a solution containing 101.2 mug per 1ml to obtain the final product.
Chromatographic conditions are as follows: the mobile phase was methanol-acetonitrile-0.1% phosphoric acid, gradient elution, elution ratios such as the following table:
the chromatographic column is Waters XbridgeTMA Shield RP18 column; the detection wavelength is 239nm, and the flow rate is 0.5ml/min；
The determination method comprises the following steps: precisely sucking 1 mu L of each of the reference solution and the sample solution, injecting into an ultra-high liquid chromatograph, measuring, and recording a chromatogram.
The fingerprint spectrum measuring method of the invention is also preferably as follows:
preparing a test solution: taking 0.2 g of the traditional Chinese medicine composition, precisely weighing, placing in a conical flask with a plug, precisely adding 20mL of 90% methanol, weighing, ultrasonically treating for 40 minutes, cooling, weighing again, supplementing the lost weight with 90% methanol, shaking uniformly, filtering, and taking the subsequent filtrate to obtain the traditional Chinese medicine composition;
preparation of control solutions: respectively weighing appropriate amounts of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, iso-forsythiaside A, forsythiaside, 4, 5-di-O-caffeoylquinic acid, quercetin and ammonium glycyrrhizinate as control substances, and preparing a mixed control solution containing 18.4 mug/ml neochlorogenic acid, 27.2 mug/ml chlorogenic acid, 21.2 mug/ml cryptochlorogenic acid, 27.2 mug/ml iso-forsythiaside A, 21.6 mug/ml forsythiaside A, 16.0 mug/ml forsythiaside, 14.4 mug/ml 4, 5-di-O-caffeoylquinic acid, 9.0 mug/ml quercetin and 24.4 mug/ml ammonium glycyrrhizinate respectively to obtain the control substance solution;
preparation of reference solutions: taking a proper amount of forsythoside A reference substance, precisely weighing, and adding 50% methanol to prepare a solution containing 101.2 mug per 1ml to obtain the final product.
Chromatographic conditions are as follows: the mobile phase was methanol-acetonitrile-0.1% phosphoric acid, gradient elution, elution ratios such as the following table:
the chromatographic column is Phenomenex Kinetx XB-C18; the detection wavelength is 239nm, and the flow rate is 0.4 ml/min;
the determination method comprises the following steps: precisely sucking 6 muL of each of the reference solution and the sample solution, injecting into an ultra-high liquid chromatograph, measuring, and recording a chromatogram.
The feasibility of the fingerprint spectrum determination method of the traditional Chinese medicine composition is evaluated from multiple aspects by using the sample prepared in example 1, and the evaluation method is as follows:
1. instruments, reagents and reagents
1.1 instruments
Waters UPLC H-CLASS ultra high performance liquid chromatograph (PDA detector, Empower 3.0 chromatographic workstation), circulating multi-purpose vacuum pump (Zheng Changchengchang department industry and trade Co., Ltd.), KQ250DB type ultrasonic cleaner (Kunshan ultrasonic instruments Co., Ltd.), Switzerland METTLER TOLEDOAL204 type electronic analytical balance, Switzerland METTLER TOLEDOAB135-S type electronic analytical balance, Shanghai Jing Ke JA2603B electronic balance.
1.2 reagents
Forsythin (China food and drug testing institute, lot No. 110821-201213, purity 95.3%), forsythoside A (China food and drug testing institute, lot No. 111810-201304, purity 94.3%), neochlorogenic acid (Chengdu Purui method science and technology development Co., Ltd., lot No. 13112712, purity 98%), chlorogenic acid (China food and drug testing institute, lot No. 110753-201314, purity 96.6%), cryptochlorogenic acid (Shanghai-sourced leaf Biotechnology Co., Ltd., lot No. ZF0226BA14, purity 98%), forsythoside A (Shanghai-sourced leaf Biotechnology Co., Ltd., No. YA0817HB14, purity 98%), 4, 5-di-O-caffeoylquinic acid (China food and drug testing institute, lot No. 111894-201102, purity 94.1%), quercetin (HWI ANALYTIK solutions, HW 12), purity, 91.7%), ammonium glycyrrhizinate (Council of Europe-EDQM, lot number: batch: 1.1, purity not less than 98%), methanol (analytically pure, Tianjin, Kogyo-Tech, batch No. 2014 1/13 days), phosphoric acid (analytically pure, Tianjin, Kogyo-Tech, batch No. 2014 3/26 days), acetonitrile (chromatographically pure, Fisher, USA, LOT 106246).
1.3 reagent
The Chinese medicinal composition of the invention (Shijiazhuang Shaling Ling pharmaceutical products Co., Ltd.)
2. Chromatographic condition optimization
2.1. Selection of detection wavelength
The chromatograms of the test solution under different detection wavelengths are examined at 210 nm-400 nm, the information content under 239nm is rich, so that the detection wavelength is optimally selected to be 239nm, and the specific information is shown in the following figures 1-4.
2.2 selection of mobile phase System
This study compared three mobile phase systems of methanol-0.1% phosphoric acid, acetonitrile-0.1% phosphoric acid and methanol-acetonitrile-0.1% phosphoric acid. The results show that the chromatographic peak of the methanol-acetonitrile-0.1% phosphoric acid system has the best resolution, and the specific information is shown in the attached figures 5-7, therefore, the mobile phase system is determined to be methanol-acetonitrile-0.1% phosphoric acid, the gradient elution is shown in the table 1, and the flow rate is 0.3 ml/min.
TABLE 1 mobile phase gradient elution Table
3. Preparation of test solution and control solution
3.1 examination of extraction solvent
The extraction effects of methanol, 90% methanol, 70% methanol and 50% methanol are respectively compared, and the result shows that the chromatographic peak in the fingerprint obtained by 50% methanol extraction is the most, and the specific information is shown in the attached figures 8-11, so that 50% methanol is selected as the extraction solvent.
3.2 examination of extraction method
The ultrasonic extraction method and the reflux extraction method are compared respectively, the number and the intensity of the chromatographic peaks of the fingerprint obtained by the ultrasonic extraction method and the reflux extraction method are not obviously different, the specific information is shown in figures 12-13, and the ultrasonic extraction method is selected because the ultrasonic extraction method is simple and convenient.
3.3 ultrasonic temporal investigation
The results of 20 minutes, 30 minutes and 40 minutes of ultrasound are respectively compared, and the results show that the chromatographic peak information amount in the fingerprint obtained by 30 minutes and 40 minutes of ultrasound is large and has no obvious difference, and the specific information is shown in figures 14-16, so that the ultrasound time is finally determined to be 30 minutes.
3.4 determination of the preparation method of the test solution: taking a proper amount of the content of the traditional Chinese medicine composition, grinding, taking 0.25g, precisely weighing, placing in a conical flask with a plug, precisely adding 25mL of 50% methanol, weighing, carrying out ultrasonic treatment (power 500W and frequency 40kHz) for 30 minutes, cooling, weighing again, complementing the weight loss by 50% methanol, shaking up, filtering, and taking the subsequent filtrate to obtain the traditional Chinese medicine composition.
3.5 preparation of control solutions: respectively weighing appropriate amounts of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, iso-forsythiaside A, forsythiaside, 4, 5-di-O-caffeoylquinic acid, quercetin and ammonium glycyrrhizinate as control substances, and preparing a mixed control solution containing 18.4 mug/ml neochlorogenic acid, 27.2 mug/ml chlorogenic acid, 21.2 mug/ml cryptochlorogenic acid, 27.2 mug/ml iso-forsythiaside A, 21.6 mug/ml forsythiaside A, 16.0 mug/ml forsythiaside, 14.4 mug/ml 4, 5-di-O-caffeoylquinic acid, 9.0 mug/ml quercetin and 24.4 mug/ml ammonium glycyrrhizinate.
3.6 preparation of reference solutions: taking a proper amount of forsythoside A reference substance, precisely weighing, and adding 50% methanol to prepare a solution containing 101.2 mug per 1ml to obtain the final product.
Spectral characterization and identification of primary chromatographic peaks
Under the determined chromatographic condition, sample introduction is respectively carried out on the reference solution and the test solution, and 16 main chromatographic peaks are determined in the chromatogram of the test solution. According to the retention time and spectral characteristics of chromatographic peaks, the peak 2 in the fingerprint is determined to be neochlorogenic acid, the peak 3 is chlorogenic acid, the peak 5 is cryptochlorogenic acid, the peak 8 is iso-forsythoside A, the peak 11 is forsythoside A, the peak 13 is quercitrin, the peak 14 is 4, 5-di-O-caffeoylquinic acid, the peak 15 is forsythin, and the peak 16 is glycyrrhizic acid, and the chromatogram and the spectrogram of the reference substance and the test sample are shown in attached figures 17-28.
Precision degree
Sample introduction is continuously carried out on the sample solution for 6 times according to the determined chromatographic conditions, the RSD of the peak area of each main chromatographic peak is less than 5 percent, the sample introduction precision is good, and the result is shown in table 2.
Repeatability of
6 parts of the content of the traditional Chinese medicine composition are taken, prepared according to the determined preparation method of the test solution, injected into a liquid chromatograph for detection, and the RSD of the peak area of each main chromatographic peak is less than 5 percent, which indicates that the method has good repeatability, and the result is shown in Table 3.
Stability of
Sample introduction is carried out on the test solution for 0h, 2h, 4h, 8h, 12h and 24h respectively, and the RSD of the peak area of each main chromatographic peak is determined to be less than 10%, which shows that the test solution is stable in 24h, and the results are shown in Table 4.
Establishment of fingerprint and similarity analysis
8.1 establishing a fingerprint: taking 10 batches of samples of the traditional Chinese medicine composition, preparing a test solution according to the requirements of 3.4 items, carrying out sample injection analysis under the chromatographic conditions to obtain the fingerprints of the 10 batches of the traditional Chinese medicine composition, adopting software 2004A version of a traditional Chinese medicine chromatogram fingerprint similarity evaluation system developed by the State pharmacopoeia Committee to analyze the fingerprints of the 10 batches of samples, carrying out multi-point correction and automatic matching on each fingerprint chromatogram peak by adopting a median method, calculating the similarity and generating a comparison fingerprint, and referring to the attached drawings 29-30, wherein the similarity result is shown in Table 5, 16 common peaks in the fingerprint are identified by standard products, and 9 common peaks in the fingerprint are identified by the standard products: neochlorogenic acid (peak 2), chlorogenic acid (peak 3), cryptochlorogenic acid (peak 5), iso-forsythoside a (peak 8), forsythoside a (peak 11), quercetin (peak 13), 4, 5-di-O-caffeoylquinic acid (peak 14), phillyrin (peak 15), glycyrrhizic acid (peak 16). Wherein, the No. 11 chromatographic peak forsythoside A with stable peak appearance and larger peak area is taken as a reference peak:
the invention establishes a liquid phase fingerprint method of the traditional Chinese medicine composition, the total chromatographic peaks are 16, and 9 common peaks are identified by a standard substance: neochlorogenic acid (peak 2), chlorogenic acid (peak 3), cryptochlorogenic acid (peak 5), iso-forsythoside a (peak 8), forsythoside a (peak 11), quercetin (peak 13), 4, 5-di-O-caffeoylquinic acid (peak 14), phillyrin (peak 15), glycyrrhizic acid (peak 16). Wherein, the 11 # chromatographic peak forsythoside A with stable peak appearance and larger peak area is taken as a reference peak, and the similarity of fingerprint spectrums of 10 batches of the traditional Chinese medicine composition is higher. The experimental results show that the method has good precision, stability and repeatability, and provides a new method for improving the quality control of the traditional Chinese medicine composition.
Drawings
FIG. 1: chromatogram of test solution with detection wavelength of 320nm
FIG. 2: chromatogram of test solution with detection wavelength of 290nm
FIG. 3: chromatogram of test solution with detection wavelength of 254nm
FIG. 4: chromatogram of test solution with detection wavelength of 230nm
FIG. 5: chromatogram of methanol-0.1% phosphoric acid sample solution
FIG. 6: chromatogram of acetonitrile-0.1% phosphoric acid sample solution
FIG. 7: chromatogram map of methanol-acetonitrile-0.1% phosphoric acid sample solution
FIG. 8: chromatogram of sample solution extracted from 50% methanol
FIG. 9: chromatogram of 70% methanol extraction test solution
FIG. 10: chromatogram of sample solution extracted from 90% methanol
FIG. 11: chromatogram of methanol extraction test solution
FIG. 12: ultrasonic 30-minute chromatogram of test solution
FIG. 13: chromatogram of test solution after refluxing for 1 hour
FIG. 14: ultrasonic 20-minute chromatogram of test solution
FIG. 15: ultrasonic 30-minute chromatogram of test solution
FIG. 16: ultrasonic 40-minute chromatogram of test solution
FIG. 17: chromatogram of control solution
FIG. 18: chromatogram of forsythoside A reference solution
FIG. 19: chromatogram of test solution
FIG. 20: comparing the new chlorogenic acid spectrogram in the reference solution with the new chlorogenic acid spectrogram in the test solution, wherein the dotted line is the test solution spectrogram, and the solid line is the reference spectrogram
FIG. 21: comparing the chlorogenic acid spectrograms of the reference solution and the test solution, the dotted line is the spectrogram of the test solution, and the solid line is the spectrogram of the reference solution
FIG. 22: comparing the spectra of the chlorogenic acid in the reference solution and the sample solution, wherein the dotted line is the spectrum of the sample, and the solid line is the spectrum of the reference
FIG. 23: comparing the reference solution with the reference solution to obtain a spectrogram of isoforsythoside A, wherein the dotted line is the spectrogram of the sample, and the solid line is the spectrogram of the reference
FIG. 24: comparing the reference solution with the sample solution to obtain forsythoside A spectrogram, wherein the dotted line is the sample spectrogram, and the solid line is the reference spectrogram
FIG. 25: comparing the spectra of quercetin in the reference solution and the test solution, wherein the dotted line is the spectrum of the test solution, and the solid line is the spectrum of the reference solution
FIG. 26: comparing the 4, 5-di-O-caffeoylquinic acid spectrograms in the reference solution and the test solution, the dotted line is the test solution spectrogram, and the solid line is the reference spectrogram
FIG. 27 is a schematic view showing: comparing the spectrograms of phillyrin in the reference solution and the test solution, the dotted line is the spectrogram of the test solution, and the solid line is the spectrogram of the reference solution
FIG. 28: comparing the glycyrrhizic acid spectrograms of the reference solution and the test solution, wherein the dotted line is the spectrogram of the test solution, and the solid line is the spectrogram of the reference solution
FIG. 29: 10 batches of the traditional Chinese medicine composition of the invention have chromatographic fingerprint
FIG. 30: the traditional Chinese medicine composition of the invention is compared with fingerprint
FIG. 31: example 1 fingerprint, wherein peak 1 in the a-diagram is neochlorogenic acid, 2 is chlorogenic acid, 3 is cryptochlorogenic acid, 4 is iso-forsythoside a, 5 is forsythoside a, 6 is quercetin, 7 is 4, 5-di-O-caffeoylquinic acid, 8 is forsythin, and 9 is glycyrrhizic acid; b is chromatogram of forsythoside A reference solution; c is the fingerprint chromatogram of the test solution
FIG. 32: example 2 fingerprint, wherein peak 1 in the a-diagram is neochlorogenic acid, 2 is chlorogenic acid, 3 is cryptochlorogenic acid, 4 is forsythiaside a, 5 is forsythiaside a, 6 is quercetin, 7 is 4, 5-di-O-caffeoylquinic acid, 8 is forsythin, and 9 is glycyrrhizic acid; b is chromatogram of forsythoside A reference solution; c is the fingerprint chromatogram of the test solution
FIG. 33: example 3 fingerprint, wherein peak 1 in the a-diagram is neochlorogenic acid, 2 is chlorogenic acid, 3 is cryptochlorogenic acid, 4 is forsythiaside a, 5 is forsythiaside a, 6 is quercetin, 7 is 4, 5-di-O-caffeoylquinic acid, 8 is forsythin, 9 is glycyrrhizic acid; b is chromatogram of forsythoside A reference solution; c is the fingerprint chromatogram of the test solution
FIG. 34: example 4 fingerprint, wherein peak 1 in the a-diagram is neochlorogenic acid, 2 is chlorogenic acid, 3 is cryptochlorogenic acid, 4 is iso-forsythoside a, 5 is forsythoside a, 6 is quercetin, 7 is 4, 5-di-O-caffeoylquinic acid, 8 is forsythin, and 9 is glycyrrhizic acid; b is chromatogram of forsythoside A reference solution; c is the fingerprint chromatogram of the test solution.
Detailed Description
Example 1
Weighing in proportion: 200g of fructus forsythiae, 300g of honeysuckle, 200g of isatis root, 40g of rhubarb, 60g of patchouli, 300g of male fern rhizome, 100g of rhodiola rosea, 9g of menthol, 60g of ephedra, 100g of bitter almond, 200g of houttuynia cordata, 100g of liquorice and 200g of gypsum, and the extraction is carried out according to the following processes:
(1) weighing the traditional Chinese medicinal materials according to the weight proportion of the raw materials, cleaning, and cutting off the traditional Chinese medicinal materials as appropriate;
(2) crushing herba Agastaches, extracting volatile oil with 10 times of water for 8 hr, and collecting volatile oil; filtering the extractive solution, discarding residue, and collecting filtrate;
(3) extracting fructus forsythiae, herba Ephedrae, herba Houttuyniae, and radix et rhizoma Rhei with 12 times of 70% ethanol for 3 times, each for 2.5 hr, mixing extractive solutions, filtering, and recovering ethanol to obtain filtrate;
(4) adding 12 times of water into honeysuckle, gypsum, isatis root, male fern rhizome, liquorice and rhodiola rosea, decocting until boiling, adding bitter apricot seed, decocting for 2 times, each time for 1 hour, combining extracting solutions, filtering, combining obtained filtrate with filtrate obtained after oil extraction of patchouli in the step (2), concentrating into clear paste with the relative density of 1.10-1.15 measured at the temperature of 60 ℃, adding ethanol, adjusting the alcohol concentration to 70%, refrigerating, standing, filtering, and recovering the ethanol until no alcohol smell exists, thus obtaining the clear paste for later use;
(5) mixing the fluid extract obtained in step (4) with the ethanol extract obtained in step (3), concentrating to obtain fluid extract with relative density of 1.15-1.20 at 60 deg.C, and drying to obtain dry extract powder;
(6) adding a proper amount of pharmaceutically acceptable auxiliary materials into the dry paste powder obtained in the step (5) for granulation;
(7) and (3) adding the menthol and the volatile oil obtained in the step (2) into ethanol for dissolving, spraying the particles obtained in the step (6), and filling into capsules.
The fingerprint spectrum measuring method comprises the following steps:
preparing a test solution: taking 0.25g of the traditional Chinese medicine composition, precisely weighing, placing in a conical flask with a plug, precisely adding 25mL of 50% methanol, weighing, ultrasonic power 500W, frequency 40kHz, ultrasonic for 30 minutes, cooling, weighing again, supplementing the lost weight with 50% methanol, shaking up, filtering, and taking the subsequent filtrate to obtain the traditional Chinese medicine composition;
preparation of control solutions: respectively weighing appropriate amounts of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, iso-forsythiaside A, forsythiaside, 4, 5-di-O-caffeoylquinic acid, quercetin and ammonium glycyrrhizinate as control substances, and preparing a mixed control solution containing 18.4 mug/ml neochlorogenic acid, 27.2 mug/ml chlorogenic acid, 21.2 mug/ml cryptochlorogenic acid, 27.2 mug/ml iso-forsythiaside A, 21.6 mug/ml forsythiaside A, 16.0 mug/ml forsythiaside, 14.4 mug/ml 4, 5-di-O-caffeoylquinic acid, 9.0 mug/ml quercetin and 24.4 mug/ml ammonium glycyrrhizinate respectively to obtain the control substance solution;
preparation of reference solutions: taking a proper amount of forsythoside A reference substance, precisely weighing, and adding 50% methanol to prepare a solution containing 101.2 mug per 1ml to obtain the final product.
Chromatographic conditions are as follows: the mobile phase was methanol-acetonitrile-0.1% phosphoric acid, gradient elution, elution ratios such as the following table:
the chromatographic column is a Waters Acquity UPLC HSS T3 chromatographic column, the column length is 100mm, the inner diameter is 2.1mm, and the particle size is 1.8 mu m; the detection wavelength is 239nm, and the flow rate is 0.3 ml/min;
the determination method comprises the following steps: precisely sucking 2 mu L of each of the reference solution and the sample solution, injecting into an ultra-high liquid chromatograph, measuring, and recording a chromatogram.
And (4) conclusion: the fingerprint spectrum is shown in figure 31, and the result is satisfactory and can be used for controlling the quality of the traditional Chinese medicine composition.
Example 2
Weighing in proportion: 300g of fructus forsythiae, 200g of honeysuckle, 300g of radix isatidis, 60g of rheum officinale, 100g of pogostemon cablin, 200g of male fern rhizome, 60g of rhodiola rosea, 5g of menthol, 100g of ephedra, 60g of bitter apricot seed, 300g of houttuynia cordata, 60g of liquorice and 300g of gypsum, and the extraction is carried out according to the following processes:
(1) weighing the traditional Chinese medicinal materials according to the weight proportion of the raw materials, cleaning, and cutting off the traditional Chinese medicinal materials as appropriate;
(2) crushing herba Agastaches, extracting volatile oil with 10 times of water for 8 hr, and collecting volatile oil; filtering the extractive solution, discarding residue, and collecting filtrate;
(3) extracting fructus forsythiae, herba Ephedrae, herba Houttuyniae, and radix et rhizoma Rhei with 12 times of 70% ethanol for 3 times, each for 2.5 hr, mixing extractive solutions, filtering, and recovering ethanol to obtain filtrate;
(4) adding 12 times of water into honeysuckle, gypsum, isatis root, male fern rhizome, liquorice and rhodiola rosea, decocting until boiling, adding bitter apricot seed, decocting for 2 times, each time for 1 hour, combining extracting solutions, filtering, combining obtained filtrate with filtrate obtained after oil extraction of patchouli in the step (2), concentrating into clear paste with the relative density of 1.10-1.15 measured at the temperature of 60 ℃, adding ethanol, adjusting the alcohol concentration to 70%, refrigerating, standing, filtering, and recovering the ethanol until no alcohol smell exists, thus obtaining the clear paste for later use;
(5) mixing the fluid extract obtained in step (4) with the ethanol extract obtained in step (3), concentrating to obtain fluid extract with relative density of 1.15-1.20 at 60 deg.C, and drying to obtain dry extract powder;
(6) adding a proper amount of pharmaceutically acceptable auxiliary materials into the dry paste powder obtained in the step (5) for granulation;
(7) and (3) adding the menthol and the volatile oil obtained in the step (2) into ethanol for dissolving, spraying the granules obtained in the step (6), and tabletting to obtain the menthol-containing tablet.
The fingerprint spectrum measuring method comprises the following steps:
preparing a test solution: taking 0.1 g of the traditional Chinese medicine composition, precisely weighing, placing in a conical flask with a plug, precisely adding 50mL of 100% methanol, weighing, ultrasonically treating for 40 minutes, cooling, weighing again, supplementing the lost weight with 100% methanol, shaking uniformly, filtering, and taking the subsequent filtrate to obtain the traditional Chinese medicine composition;
preparation of control solutions: respectively weighing appropriate amounts of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, iso-forsythiaside A, forsythiaside, 4, 5-di-O-caffeoylquinic acid, quercetin and ammonium glycyrrhizinate as control substances, and preparing a mixed control solution containing 18.4 mug/ml neochlorogenic acid, 27.2 mug/ml chlorogenic acid, 21.2 mug/ml cryptochlorogenic acid, 27.2 mug/ml iso-forsythiaside A, 21.6 mug/ml forsythiaside A, 16.0 mug/ml forsythiaside, 14.4 mug/ml 4, 5-di-O-caffeoylquinic acid, 9.0 mug/ml quercetin and 24.4 mug/ml ammonium glycyrrhizinate respectively to obtain the control substance solution;
preparation of reference solutions: taking a proper amount of forsythoside A reference substance, precisely weighing, and adding 50% methanol to prepare a solution containing 101.2 mug per 1ml to obtain the final product.
Chromatographic conditions are as follows: the mobile phase was methanol-acetonitrile-0.1% phosphoric acid, gradient elution, elution ratios such as the following table:
the chromatographic column is a Waters Acquity UPLC HSS T3 (column length is 100mm, inner diameter is 2.1mm, particle size is 1.8 mu m) chromatographic column; the detection wavelength is 239nm, and the flow rate is 0.1 ml/min;
the determination method comprises the following steps: precisely sucking 1 mu L of each of the reference solution and the sample solution, injecting into an ultra-high liquid chromatograph, measuring, and recording a chromatogram.
And (4) conclusion: the fingerprint spectrum is shown in figure 32, and the result is satisfactory and can be used for controlling the quality of the traditional Chinese medicine composition.
Example 3
The formula of the raw material medicine is as follows: 278g of fructus forsythiae, 294g of honeysuckle, 285g of isatis root, 55g of rhubarb, 95g of patchouli, 290g of male fern rhizome, 87g of rhodiola rosea, 8.5g of menthol, 88g of ephedra, 80g of bitter almond, 284g of houttuynia cordata, 95g of liquorice and 277g of gypsum, and the extraction is carried out according to the following processes:
(1) weighing the traditional Chinese medicinal materials according to the weight proportion of the raw materials, cleaning, and cutting off the traditional Chinese medicinal materials as appropriate;
(2) crushing herba Agastaches, extracting volatile oil with 10 times of water for 8 hr, and collecting volatile oil; filtering the extractive solution, discarding residue, and collecting filtrate;
(3) extracting fructus forsythiae, herba Ephedrae, herba Houttuyniae, and radix et rhizoma Rhei with 12 times of 70% ethanol for 3 times, each for 2.5 hr, mixing extractive solutions, filtering, and recovering ethanol to obtain filtrate;
(4) adding 12 times of water into honeysuckle, gypsum, isatis root, male fern rhizome, liquorice and rhodiola rosea, decocting until boiling, adding bitter apricot seed, decocting for 2 times, each time for 1 hour, combining extracting solutions, filtering, combining obtained filtrate with filtrate obtained after oil extraction of patchouli in the step (2), concentrating into clear paste with the relative density of 1.10-1.15 measured at the temperature of 60 ℃, adding ethanol, adjusting the alcohol concentration to 70%, refrigerating, standing, filtering, and recovering the ethanol until no alcohol smell exists, thus obtaining the clear paste for later use;
(5) mixing the fluid extract obtained in step (4) with the ethanol extract obtained in step (3), concentrating to obtain fluid extract with relative density of 1.15-1.20 at 60 deg.C, and drying to obtain dry extract powder;
(6) adding a proper amount of pharmaceutically acceptable auxiliary materials into the dry paste powder obtained in the step (5) for granulation;
(7) and (3) adding the menthol and the volatile oil obtained in the step (2) into ethanol for dissolving, spraying the granules obtained in the step (6), and bagging.
The fingerprint spectrum measuring method comprises the following steps:
preparing a test solution: taking 0.5 g of the traditional Chinese medicine composition, precisely weighing, placing in a conical flask with a plug, precisely adding 10mL of 40% methanol, weighing, ultrasonically treating for 20 minutes, cooling, weighing again, supplementing the lost weight with 40% methanol, shaking uniformly, filtering, and taking the subsequent filtrate to obtain the traditional Chinese medicine composition;
preparation of control solutions: respectively weighing appropriate amounts of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, iso-forsythiaside A, forsythiaside, 4, 5-di-O-caffeoylquinic acid, quercetin and ammonium glycyrrhizinate as control substances, and preparing a mixed control solution containing 18.4 mug/ml neochlorogenic acid, 27.2 mug/ml chlorogenic acid, 21.2 mug/ml cryptochlorogenic acid, 27.2 mug/ml iso-forsythiaside A, 21.6 mug/ml forsythiaside A, 16.0 mug/ml forsythiaside, 14.4 mug/ml 4, 5-di-O-caffeoylquinic acid, 9.0 mug/ml quercetin and 24.4 mug/ml ammonium glycyrrhizinate respectively to obtain the control substance solution;
preparation of reference solutions: taking a proper amount of forsythoside A reference substance, precisely weighing, and adding 50% methanol to prepare a solution containing 101.2 mug per 1ml to obtain the final product.
Chromatographic conditions are as follows: the mobile phase was methanol-acetonitrile-0.1% phosphoric acid, gradient elution, elution ratios such as the following table:
the chromatographic column is Waters XbridgeTMShield RP18 (5 μm, 4.6 mm × 250 mm) chromatography column; the detection wavelength is 239nm, and the flow rate is 0.5 ml/min;
the determination method comprises the following steps: precisely sucking 1 mu L of each of the reference solution and the sample solution, injecting into an ultra-high liquid chromatograph, measuring, and recording a chromatogram.
As a result: the fingerprint spectrum is shown in figure 33, and the result is satisfactory and can be used for controlling the quality of the traditional Chinese medicine composition.
Example 4
The formula of the raw material medicine is as follows: weighing in proportion: 255g of fructus forsythiae, 255g of honeysuckle, 255g of isatis root, 51g of rhubarb, 85g of patchouli, 255g of male fern rhizome, 85g of rhodiola rosea, 7.5g of menthol, 85g of ephedra, 85g of bitter almond, 255g of cordate houttuynia, 85g of liquorice and 255g of gypsum, and the extraction is carried out according to the following processes:
(1) weighing the traditional Chinese medicinal materials according to the weight proportion of the raw materials, cleaning, and cutting off the traditional Chinese medicinal materials as appropriate;
(2) crushing herba Agastaches, extracting volatile oil with 10 times of water for 8 hr, and collecting volatile oil; filtering the extractive solution, discarding residue, and collecting filtrate;
(3) extracting fructus forsythiae, herba Ephedrae, herba Houttuyniae, and radix et rhizoma Rhei with 12 times of 70% ethanol for 3 times, each for 2.5 hr, mixing extractive solutions, filtering, and recovering ethanol to obtain filtrate;
(4) adding 12 times of water into honeysuckle, gypsum, isatis root, male fern rhizome, liquorice and rhodiola rosea, decocting until boiling, adding bitter apricot seed, decocting for 2 times, each time for 1 hour, combining extracting solutions, filtering, combining obtained filtrate with filtrate obtained after oil extraction of patchouli in the step (2), concentrating into clear paste with the relative density of 1.10-1.15 measured at the temperature of 60 ℃, adding ethanol, adjusting the alcohol concentration to 70%, refrigerating, standing, filtering, and recovering the ethanol until no alcohol smell exists, thus obtaining the clear paste for later use;
(5) mixing the fluid extract obtained in step (4) with the ethanol extract obtained in step (3), concentrating to obtain fluid extract with relative density of 1.15-1.20 at 60 deg.C, and drying to obtain dry extract powder;
(6) adding a proper amount of pharmaceutically acceptable auxiliary materials into the dry paste powder obtained in the step (5) for granulation;
(7) and (3) adding the menthol and the volatile oil obtained in the step (2) into ethanol for dissolving, spraying the particles obtained in the step (6), and filling into capsules.
The fingerprint spectrum measuring method comprises the following steps:
preparing a test solution: taking 0.2 g of the traditional Chinese medicine composition, precisely weighing, placing in a conical flask with a plug, precisely adding 20mL of 90% methanol, weighing, ultrasonically treating for 40 minutes, cooling, weighing again, supplementing the lost weight with 90% methanol, shaking uniformly, filtering, and taking the subsequent filtrate to obtain the traditional Chinese medicine composition;
preparation of control solutions: respectively weighing appropriate amounts of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, iso-forsythiaside A, forsythiaside, 4, 5-di-O-caffeoylquinic acid, quercetin and ammonium glycyrrhizinate as control substances, and preparing a mixed control solution containing 18.4 mug/ml neochlorogenic acid, 27.2 mug/ml chlorogenic acid, 21.2 mug/ml cryptochlorogenic acid, 27.2 mug/ml iso-forsythiaside A, 21.6 mug/ml forsythiaside A, 16.0 mug/ml forsythiaside, 14.4 mug/ml 4, 5-di-O-caffeoylquinic acid, 9.0 mug/ml quercetin and 24.4 mug/ml ammonium glycyrrhizinate respectively to obtain the control substance solution;
preparation of reference solutions: taking a proper amount of forsythoside A reference substance, precisely weighing, and adding 50% methanol to prepare a solution containing 101.2 mug per 1ml to obtain the final product.
Chromatographic conditions are as follows: the mobile phase was methanol-acetonitrile-0.1% phosphoric acid, gradient elution, elution ratios such as the following table:
the chromatographic column is a Phenomenex Kinetx XB-C18 chromatographic column; the detection wavelength is 239nm, and the flow rate is 0.4 ml/min;
the determination method comprises the following steps: precisely sucking 6 muL of each of the reference solution and the sample solution, injecting into an ultra-high liquid chromatograph, measuring, and recording a chromatogram.
And (4) conclusion: the fingerprint spectrum is shown in figure 34, and the result is satisfactory and can be used for controlling the quality of the traditional Chinese medicine composition.
Claims (6)
1. A fingerprint spectrum measuring method of a traditional Chinese medicine composition is disclosed, the traditional Chinese medicine composition is prepared from the following raw material medicines in parts by weight: 200-300 parts of fructus forsythiae, 60-100 parts of ephedra, 40-60 parts of rheum officinale, 200-300 parts of houttuynia cordata, 200-300 parts of honeysuckle, 200-300 parts of radix isatidis, 60-100 parts of pogostemon cablin, 200-300 parts of rhizoma dryopteris crassirhizomae, 60-100 parts of rhodiola rosea, 5-9 parts of menthol, 60-100 parts of bitter almond, 60-100 parts of liquorice and 200-300 parts of gypsum, and is characterized in that the fingerprint spectrum determination method is as follows:
preparing a test solution: taking 0.25g of the traditional Chinese medicine composition, precisely weighing, placing in a conical flask with a plug, precisely adding 25mL of 50% methanol, weighing, ultrasonic power 500W, frequency 40kHz, ultrasonic for 30 minutes, cooling, weighing again, supplementing the lost weight with 50% methanol, shaking up, filtering, and taking the subsequent filtrate to obtain the traditional Chinese medicine composition;
preparation of control solutions: respectively weighing appropriate amounts of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, iso-forsythiaside A, forsythiaside, 4, 5-di-O-caffeoylquinic acid, quercetin and ammonium glycyrrhizinate as control substances, and preparing a mixed control solution containing 18.4 mug/ml neochlorogenic acid, 27.2 mug/ml chlorogenic acid, 21.2 mug/ml cryptochlorogenic acid, 27.2 mug/ml iso-forsythiaside A, 21.6 mug/ml forsythiaside A, 16.0 mug/ml forsythiaside, 14.4 mug/ml 4, 5-di-O-caffeoylquinic acid, 9.0 mug/ml quercetin and 24.4 mug/ml ammonium glycyrrhizinate respectively to obtain the control substance solution;
preparation of reference solutions: taking a proper amount of forsythiaside A reference substance, precisely weighing, and adding 50% methanol to prepare a solution containing 101.2 microgram per 1ml to obtain the forsythiaside A;
the mobile phase was methanol-acetonitrile-0.1% phosphoric acid, gradient elution, elution ratios such as the following table:
the chromatographic column is a Waters Acquity UPLC HSS T3 chromatographic column, the column length is 100mm, the inner diameter is 2.1mm, and the particle size is 1.8 mu m; the detection wavelength is 239nm, and the flow rate is 0.3 ml/min;
the determination method comprises the following steps: precisely sucking 2 mu L of each of the reference solution and the sample solution, injecting into an ultra-high liquid chromatograph, measuring, and recording a chromatogram.
2. The fingerprint spectrum measuring method according to claim 1, wherein the traditional Chinese medicine composition is prepared from the following raw material medicines in parts by weight:
200 parts of fructus forsythiae, 300 parts of honeysuckle, 200 parts of isatis root, 40 parts of rhubarb, 60 parts of cablin potchouli herb, 300 parts of male fern rhizome, 100 parts of rhodiola rosea, 9 parts of menthol, 60 parts of ephedra, 100 parts of bitter apricot seed, 200 parts of cordate houttuynia, 100 parts of liquorice and 200 parts of gypsum.
3. The fingerprint spectrum measuring method according to claim 1, wherein the Chinese medicinal composition is prepared from the following raw material medicines in parts by weight:
300 parts of fructus forsythiae, 200 parts of honeysuckle, 300 parts of isatis root, 60 parts of rheum officinale, 100 parts of pogostemon cablin, 200 parts of male fern rhizome, 60 parts of rhodiola rosea, 5 parts of menthol, 100 parts of ephedra, 60 parts of bitter apricot seed, 300 parts of houttuynia cordata, 60 parts of liquorice and 300 parts of gypsum.
4. The fingerprint spectrum measuring method according to claim 1, wherein the Chinese medicinal composition is prepared from the following raw material medicines in parts by weight:
fructus forsythiae 278, honeysuckle 294, isatis root 285, rhubarb 55, patchouli 95, male fern 290, rhodiola rosea 87, menthol 8.5, ephedra 88, bitter apricot kernel 80, cordate houttuynia 284, liquorice 95 and gypsum 277.
5. The fingerprint spectrum measuring method according to claim 1, wherein the Chinese medicinal composition is prepared from the following raw material medicines in parts by weight:
fructus forsythiae 255, honeysuckle 255, isatis root 255, rhubarb 51, patchouli 85, male fern 255, rhodiola rosea 85, menthol 7.5, ephedra 85, bitter apricot seed 85, cordate houttuynia 255, liquorice 85 and gypsum 255.
6. The fingerprint determination method according to any one of claims 1 to 5, wherein the preparation method of the Chinese medicinal composition comprises:
(1) weighing the traditional Chinese medicinal materials according to the weight proportion of the raw materials, cleaning, and cutting off the traditional Chinese medicinal materials as appropriate;
(2) crushing herba Agastaches, extracting volatile oil with 10 times of water for 8 hr, and collecting volatile oil; filtering the extractive solution, discarding residue, and collecting filtrate;
(3) extracting fructus forsythiae, herba Ephedrae, herba Houttuyniae, and radix et rhizoma Rhei with 12 times of 70% ethanol for 3 times, each for 2.5 hr, mixing extractive solutions, filtering, and recovering ethanol to obtain filtrate;
(4) adding 12 times of water into honeysuckle, gypsum, isatis root, male fern rhizome, liquorice and rhodiola rosea, decocting until boiling, adding bitter apricot seed, decocting for 2 times, each time for 1 hour, combining extracting solutions, filtering, combining obtained filtrate with filtrate obtained after oil extraction of patchouli in the step (2), concentrating into clear paste with the relative density of 1.10-1.15 measured at the temperature of 60 ℃, adding ethanol, adjusting the alcohol concentration to 70%, refrigerating, standing, filtering, and recovering the ethanol until no alcohol smell exists, thus obtaining the clear paste for later use;
(5) mixing the fluid extract obtained in step (4) with the ethanol extract obtained in step (3), concentrating to obtain fluid extract with relative density of 1.15-1.20 at 60 deg.C, and drying to obtain dry extract powder;
(6) adding a proper amount of pharmaceutically acceptable auxiliary materials into the dry paste powder obtained in the step (5) for granulation;
(7) and (3) dissolving the menthol and the volatile oil obtained in the step (2) in ethanol, spraying the granules obtained in the step (6), sealing, uniformly mixing, tabletting, and encapsulating or bagging.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610119793.8A CN107153098B (en) | 2016-03-03 | 2016-03-03 | Method for measuring fingerprint spectrum of traditional Chinese medicine composition |
| PCT/CN2017/075513 WO2017148426A1 (en) | 2016-03-03 | 2017-03-03 | Method for determining fingerprint of chinese medicine composition |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610119793.8A CN107153098B (en) | 2016-03-03 | 2016-03-03 | Method for measuring fingerprint spectrum of traditional Chinese medicine composition |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107153098A CN107153098A (en) | 2017-09-12 |
| CN107153098B true CN107153098B (en) | 2021-05-04 |
Family
ID=59743593
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610119793.8A Active CN107153098B (en) | 2016-03-03 | 2016-03-03 | Method for measuring fingerprint spectrum of traditional Chinese medicine composition |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN107153098B (en) |
| WO (1) | WO2017148426A1 (en) |
Families Citing this family (94)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109668970B (en) * | 2017-10-13 | 2020-11-10 | 石家庄以岭药业股份有限公司 | Ultra-high performance liquid chromatography detection method for traditional Chinese medicine composition |
| CN109776635B (en) | 2017-11-10 | 2021-08-17 | 北京以岭药业有限公司 | Method for separating eight components in traditional Chinese medicine composition |
| CN108645921B (en) * | 2018-01-24 | 2022-08-26 | 通药制药集团股份有限公司 | Method for constructing HPLC fingerprint of Chinese patent medicine' Xiaomi Shuan |
| CN108459110A (en) * | 2018-05-09 | 2018-08-28 | 国珍健康科技（北京）有限公司 | A method of chlorogenic acid is measured by high performance liquid chromatography |
| CN109187820B (en) * | 2018-11-09 | 2021-07-23 | 湖南国华制药有限公司 | Method for establishing UPLC fingerprint of compound formula chinaroot greenbrier granules |
| CN109633043A (en) * | 2019-02-27 | 2019-04-16 | 通化玉圣药业有限公司 | A kind of method for building up and its standard diagram of gadol injection HPLC finger-print |
| CN111766308B (en) * | 2019-03-28 | 2023-11-10 | 石家庄以岭药业股份有限公司 | Fingerprint spectrum measuring method for traditional Chinese medicine composition preparation |
| CN110208400B (en) * | 2019-05-21 | 2023-09-19 | 新疆农业科学院农产品贮藏加工研究所 | Method for determining phenol content of various monomers in red dates |
| CN112014481B (en) * | 2019-05-29 | 2022-10-25 | 湖北梦阳药业股份有限公司 | Detection method of raw white oral liquid fingerprint spectrum |
| CN112326806B (en) * | 2019-07-20 | 2022-11-04 | 神威药业集团有限公司 | Albizzia julibrissin formula particle HPLC characteristic spectrum and construction method and application thereof |
| CN110376305A (en) * | 2019-07-26 | 2019-10-25 | 淮阴工学院 | A kind of content assaying method of wormwood artemisia tea Content of Chlorogenic Acid class |
| CN112557563A (en) * | 2019-09-10 | 2021-03-26 | 泰州医药城国科化物生物医药科技有限公司 | Method for identifying medicinal material fructus aurantii in western and Jiangxi regions |
| CN112578029B (en) * | 2019-09-29 | 2022-10-04 | 天津同仁堂集团股份有限公司 | HPLC detection method for lung clearing decoction preparation and application thereof |
| CN110632232B (en) * | 2019-10-22 | 2021-09-24 | 山东省中医药研究院 | Method for establishing fingerprint spectrum of fructus forsythiae medicinal material |
| CN112710765A (en) * | 2019-10-24 | 2021-04-27 | 泰州医药城国科化物生物医药科技有限公司 | Fingerprint detection method of gardenia medicinal material and application thereof |
| CN110850019B (en) * | 2019-11-29 | 2022-02-18 | 南京南医堂健康产业发展有限公司 | Method for detecting fingerprint of Minggu Shuqing drink |
| CN110716001B (en) * | 2019-11-29 | 2020-07-07 | 山东汉方制药有限公司 | Method for detecting characteristic spectrum of compound phellodendron amurense liquid preparation |
| CN111323504B (en) * | 2020-03-16 | 2022-09-06 | 浙江凯润药业股份有限公司 | Yiqing granule fingerprint spectrum quality control method |
| CN113588800B (en) * | 2020-04-30 | 2024-05-17 | 河北凯盛医药科技有限公司 | Preparation and detection method of classical famous prescription Xiebai powder material standard |
| CN111983121A (en) * | 2020-07-03 | 2020-11-24 | 湖南旻一生物科技有限公司 | Quality control method and quality control spectrogram of non-volatile components of Chinese medicinal sachet, and construction method thereof |
| CN111830170A (en) * | 2020-07-24 | 2020-10-27 | 陕西康城药业股份有限公司 | Method for measuring content of antiviral mixture in four seasons |
| CN112076151B (en) * | 2020-08-28 | 2023-04-18 | 乐泰药业有限公司 | A Chinese medicinal oral liquid for treating diabetes due to deficiency of both qi and yin, and its preparation method and quality control method |
| CN112268976B (en) * | 2020-09-11 | 2022-05-20 | 浙江磐康药业有限公司 | Establishment method of HPLC fingerprint for distinguishing rubus chingii and rubus suavissimus and fingerprint |
| CN112098556B (en) * | 2020-09-21 | 2022-08-30 | 山东宏济堂制药集团股份有限公司 | Detection method of angelica sinensis Liuhuang decoction |
| CN114252511B (en) * | 2020-09-21 | 2023-05-12 | 厦门中药厂有限公司 | Method for detecting eight-treasure pill fingerprint and eight-treasure pill standard fingerprint and application |
| CN112268968B (en) * | 2020-10-16 | 2022-05-10 | 山东步长神州制药有限公司 | Detection method of fingerprint spectrum of medicinal preparation |
| CN112505170B (en) * | 2020-11-11 | 2022-03-08 | 成都中医药大学 | Qinglong white tiger soup and quality detection method thereof |
| CN112557526B (en) * | 2020-11-16 | 2022-07-19 | 成都市药品检验研究院 | Quality-controllable isatis root extract and method for identifying isatis root containing raw material medicine in Chinese patent medicine |
| CN114515301B (en) * | 2020-11-19 | 2024-03-29 | 国药集团同济堂（贵州）制药有限公司 | Semen astragali Complanati standard decoction, and preparation method and detection method thereof |
| CN112394126A (en) * | 2020-11-30 | 2021-02-23 | 天津中新药业集团股份有限公司隆顺榕制药厂 | Extraction and detection method and application of chlorogenic acid in solanum dulcamara |
| CN114636760B (en) * | 2020-12-15 | 2024-01-02 | 云南雷允上理想药业有限公司 | Fingerprint spectrum establishment method of traditional Chinese medicine composition for treating kidney diseases and application of fingerprint spectrum establishment method |
| CN114720569A (en) * | 2020-12-22 | 2022-07-08 | 云南雷允上理想药业有限公司 | Fingerprint spectrum establishment method of achyranthes bidentata medicinal material and application thereof |
| CN114689780B (en) * | 2020-12-29 | 2023-10-03 | 四川新绿色药业科技发展有限公司 | Multi-component quality detection method for Baoyin decoction extract |
| CN114689712B (en) * | 2020-12-29 | 2023-10-03 | 四川新绿色药业科技发展有限公司 | Multi-component quality detection method for channel warming soup extract |
| CN114689710B (en) * | 2020-12-29 | 2023-10-03 | 四川新绿色药业科技发展有限公司 | Multi-component quality detection method for loquat lung-heat-clearing drink extract |
| CN112834672B (en) * | 2020-12-31 | 2023-09-08 | 四川新绿色药业科技发展有限公司 | Construction and detection method of feature map of rangooncreeper fruit formula granule |
| CN112903844B (en) * | 2021-01-19 | 2022-12-16 | 皖西学院 | Detection method for content of pteridonin in pteridium aquilinum |
| CN112870376B (en) * | 2021-01-27 | 2023-07-11 | 上海硕方医药科技有限公司 | Traditional Chinese medicine extracting solution concentration defoaming method and application thereof |
| CN112946113B (en) * | 2021-02-01 | 2022-09-30 | 湖北医药学院 | Method for identifying genuine medicinal material Shiweir fructus forsythiae basal source |
| CN113960184A (en) * | 2021-02-05 | 2022-01-21 | 株洲千金药业股份有限公司 | Establishment and quality detection method of blood-replenishing and motherwort-tonifying pill fingerprint spectrum |
| CN113125628A (en) * | 2021-03-12 | 2021-07-16 | 河北锦坤动物药业有限公司 | Thin-layer chromatography identification method for radix sophorae flavescentis and radix sophorae flavescentis in three-flavor bistort oral liquid |
| CN112858529B (en) * | 2021-03-12 | 2023-07-18 | 内蒙古奥特奇蒙药股份有限公司 | Detection method of three-ingredient rheum officinale preparation |
| CN113138238A (en) * | 2021-04-15 | 2021-07-20 | 河南省纳普生物技术有限公司 | Method for detecting content of 8 flavone and phenolic acid components in camelina sativa seeds |
| CN113156017B (en) * | 2021-04-26 | 2023-02-24 | 陕西汉王药业股份有限公司 | Method for simultaneously determining contents of 12 chemical components in strong dizzy-stop tablet by adopting HPLC (high performance liquid chromatography) |
| CN113156019B (en) * | 2021-04-28 | 2022-09-30 | 重庆市畜牧科学院 | Method for detecting multi-component content of liquorice heart-fire-purging decoction |
| CN113176366A (en) * | 2021-05-08 | 2021-07-27 | 宁夏医科大学 | Evaluation method of secondary refined polysaccharide and flavonoid components based on spectral efficiency relationship |
| CN113418996A (en) * | 2021-05-13 | 2021-09-21 | 江西普正制药股份有限公司 | Callicarpa nudiflora antibacterial action substance evaluation method based on spectrum effect correlation |
| CN113376273B (en) * | 2021-06-02 | 2023-03-17 | 国药集团精方（安徽）药业股份有限公司 | Detection method of HPLC (high performance liquid chromatography) characteristic spectrum of Qingxin lotus seed drink and application of characteristic spectrum |
| CN113466361A (en) * | 2021-06-18 | 2021-10-01 | 广西壮族自治区中医药研究院 | Method for evaluating quality of gastrointestinal powder |
| CN113834883B (en) * | 2021-06-22 | 2023-05-26 | 海南葫芦娃药业集团股份有限公司 | Method for measuring content of forsythin in children's lung heat cough and asthma granule |
| CN113588808B (en) * | 2021-06-25 | 2023-02-28 | 西安千禾药业股份有限公司 | Content determination method for simultaneously detecting multiple active ingredients in Xueping capsules |
| CN113341031A (en) * | 2021-07-21 | 2021-09-03 | 青海省中医院 | Method for identifying quality goods, counterfeit goods and substitutes of rhodiola root medicinal materials |
| CN115718145A (en) * | 2021-08-24 | 2023-02-28 | 河北以岭医药研究院有限公司 | Method for measuring fingerprint spectrum of traditional Chinese medicine composition |
| CN113759038B (en) * | 2021-08-26 | 2023-04-07 | 北京康仁堂药业有限公司 | Trachelospermi caulis characteristic map, construction method and quality control method thereof, trachelospermi caulis formula granules and preparation method thereof |
| CN113791165B (en) * | 2021-09-13 | 2023-09-26 | 湖南新汇制药股份有限公司 | Quality detection method for stir-fried fructus viticis standard decoction |
| CN114414670B (en) * | 2021-10-29 | 2024-05-07 | 柳州市妇幼保健院 | Establishment method of horseshoe gold fingerprint and fingerprint thereof |
| CN114166958B (en) * | 2021-10-29 | 2024-03-29 | 合肥创新医药技术有限公司 | Detection method of fingerprint of traditional Chinese medicine compound herba xanthil stomach-calming particles and application thereof |
| CN113960210A (en) * | 2021-10-31 | 2022-01-21 | 江苏海洋大学 | Detection method of high performance liquid chromatography fingerprint of pharbitis seed |
| CN113917041B (en) * | 2021-11-01 | 2024-03-15 | 湖南新汇制药股份有限公司 | Quality detection method for cortex moutan standard decoction |
| CN114235974B (en) * | 2021-11-05 | 2024-05-31 | 新疆维吾尔药业有限责任公司 | Method for detecting rutin, isoquercitrin and rosmarinic acid content in herba Hedyotidis Diffusae or extract thereof |
| CN113960214A (en) * | 2021-11-09 | 2022-01-21 | 上海海虹实业(集团)巢湖今辰药业有限公司 | Rhizoma atractylodis macrocephalae component selection method based on rhizoma alismatis decoction |
| CN113960213A (en) * | 2021-11-09 | 2022-01-21 | 上海海虹实业(集团)巢湖今辰药业有限公司 | Alisma orientale component selection method based on alisma orientale decoction |
| CN114152686B (en) * | 2021-11-10 | 2023-11-03 | 湖南易能生物医药有限公司 | Fingerprint construction method of traditional Chinese medicine compound containing cinnamon and application of fingerprint construction method |
| CN114062553B (en) * | 2021-11-17 | 2024-05-07 | 荣昌制药(淄博)有限公司 | HPLC fingerprint determination method of children six-ingredient food-enhancing ointment |
| CN114235993B (en) * | 2021-12-02 | 2024-04-26 | 魏秀丽 | Method for detecting 13 effective components in baphicacanthus cusia antiphlogistic oral liquid by adopting HPLC-PDA (high Performance liquid chromatography-personal digital Assistant) |
| CN114184700B (en) * | 2021-12-06 | 2023-08-22 | 内蒙古民族大学 | Method for measuring chemical component content of Mongolian medicine costa |
| CN113945674B (en) * | 2021-12-20 | 2023-01-13 | 江西省药品检验检测研究院 | Characteristic spectrum and analysis method of processed rehmannia root product |
| CN114354828B (en) * | 2021-12-29 | 2023-11-03 | 甘肃奇正藏药有限公司 | Characteristic spectrum of herba Lagotis Brevibacterium, construction method thereof and quality detection method of herba Lagotis Brevibacterium |
| CN114460191A (en) * | 2022-01-20 | 2022-05-10 | 广西壮族自治区食品药品检验所 | Method for measuring content of digitoxin D in Tianwang heart tonifying preparation |
| CN114441685B (en) * | 2022-02-08 | 2023-06-27 | 湖南新汇制药股份有限公司 | Paris polyphylla standard decoction quality detection method |
| CN114609291B (en) * | 2022-03-22 | 2023-12-26 | 国药集团精方（安徽）药业股份有限公司 | Fingerprint detection method for traditional Chinese medicine compound consistent decoction |
| CN114894917B (en) * | 2022-04-06 | 2023-10-20 | 山东步长制药股份有限公司 | Method for detecting volatile components of rhizoma nardostachyos and constructing fingerprint |
| CN114755338B (en) * | 2022-04-19 | 2023-10-20 | 四川新绿色药业科技发展有限公司 | Detection method of wrinkled giant hyssop and preparation thereof |
| CN114942291B (en) * | 2022-06-20 | 2024-03-12 | 江苏省中医院 | Quality detection method of yang-suppressing yin-nourishing particles |
| CN115184493A (en) * | 2022-07-06 | 2022-10-14 | 株洲市食品药品检验所 | Fingerprint detection method for active ingredients in taxus mairei medicinal material and traditional Chinese medicine decoction pieces thereof |
| CN115032311B (en) * | 2022-07-11 | 2024-03-22 | 景忠山国药（唐山）有限公司 | Method for measuring content of Xinnaoxin medicine components based on one-measurement-multiple-evaluation method |
| CN115201376A (en) * | 2022-07-18 | 2022-10-18 | 国药集团精方（安徽）药业股份有限公司 | Quality detection method of loquat cough relieving granules |
| CN114990054B (en) * | 2022-07-22 | 2023-07-21 | 成都艾伟孚生物科技有限公司 | Sperm washing fertilization liquid for auxiliary reproduction |
| CN115372501B (en) * | 2022-07-27 | 2024-03-26 | 广州科曼生物科技有限公司 | Control extract of fritillaria thunbergii or fritillaria hubei, preparation method and application thereof |
| CN115389654B (en) * | 2022-08-04 | 2023-10-27 | 华润三九现代中药制药有限公司 | Construction method and content testing method of fingerprint of centipeda minima medicinal preparation |
| CN115343388B (en) * | 2022-08-04 | 2023-09-08 | 中山市中医院 | Method for evaluating quality of pagodatree flower particles by using one-test-multiple-evaluation method |
| CN115372510B (en) * | 2022-08-16 | 2024-03-15 | 云南植物药业有限公司 | Hypocrellin fingerprint and establishment method and application thereof |
| CN115452971A (en) * | 2022-08-17 | 2022-12-09 | 神威药业集团有限公司 | Method for analyzing chemical components of Shengyu decoction based on UPLC-Q-TOF-MS technology and identification |
| CN115327007A (en) * | 2022-08-17 | 2022-11-11 | 浙江双石药业有限公司 | Method for detecting fingerprint spectrum of ephedra decoction |
| CN115326981B (en) * | 2022-08-22 | 2024-04-30 | 贵州医科大学 | Method for measuring content of various components in garden balsam medicinal material |
| CN115343395B (en) * | 2022-08-26 | 2023-12-26 | 华颐药业有限公司 | Establishment method of UPLC fingerprint of scattered decoction and standard fingerprint and application thereof |
| CN115494173A (en) * | 2022-09-21 | 2022-12-20 | 江西济民可信药业有限公司 | Fingerprint detection method of infantile heat cough oral liquid |
| CN115452996B (en) * | 2022-09-23 | 2024-02-02 | 江苏省中医院 | Quality detection method of traditional Chinese medicine compound preparation for treating viral upper respiratory tract infection |
| CN115494181B (en) * | 2022-10-17 | 2024-05-17 | 山东省食品药品检验研究院 | Detection method for simultaneously identifying five raw materials in capsule for dredging collaterals and resolving phlegm |
| CN115684425A (en) * | 2022-11-11 | 2023-02-03 | 厦门中药厂有限公司 | Kidney tonifying and essence strengthening pill fingerprint detection method, comparison fingerprint and application |
| CN116106452A (en) * | 2023-02-27 | 2023-05-12 | 浙江景岳堂药业有限公司 | Construction method of stevia rebaudiana formula granule characteristic spectrum, characteristic spectrum constructed by method and application of characteristic spectrum |
| CN117191984A (en) * | 2023-09-06 | 2023-12-08 | 嘉亨(珠海横琴)医药科技有限公司 | Quantitative detection method of Ganweikang tablets and application thereof |
| CN117233306B (en) * | 2023-11-16 | 2024-01-30 | 江西省药品检验检测研究院 | Fingerprint establishment and component content determination method for jindan Fuyan particles |
| CN118393052A (en) * | 2024-06-24 | 2024-07-26 | 山东则正医药技术有限公司 | Detection method and application of related substances in hydroxychloroquine sulfate |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1194752C (en) * | 2003-07-01 | 2005-03-30 | 河北以岭医药研究院有限公司 | Anti-virus Chinese medicine composition and preparation process thereof |
| CN101683421B (en) * | 2008-09-26 | 2013-02-27 | 北京以岭药业有限公司 | Applications of traditional Chinese medicine composition in preparation of medicament for treating repetitive respiratory tract infection |
| CN103800523B (en) * | 2012-11-13 | 2018-09-25 | 河北以岭医药研究院有限公司 | A kind of preparation method of anti virus herb composition and the assay method of finger-print |
| CN104345111B (en) * | 2013-08-08 | 2017-04-05 | 石家庄以岭药业股份有限公司 | The assay method of various active component content in a kind of Chinese medicinal composition preparation |
-
2016
- 2016-03-03 CN CN201610119793.8A patent/CN107153098B/en active Active
-
2017
- 2017-03-03 WO PCT/CN2017/075513 patent/WO2017148426A1/en active Application Filing
Also Published As
| Publication number | Publication date |
|---|---|
| WO2017148426A1 (en) | 2017-09-08 |
| CN107153098A (en) | 2017-09-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107153098B (en) | Method for measuring fingerprint spectrum of traditional Chinese medicine composition | |
| CN107149623B (en) | Content determination method of traditional Chinese medicine composition | |
| WO2023024322A1 (en) | Method for determining fingerprint of traditional chinese medicine composition | |
| CN101991661B (en) | Method for detecting Chinese patent drug containing at least two of white paeony root, ginseng, salvia miltiorrhiza, sweet wormwood, liquorice and angelica sinensis | |
| CN104914199B (en) | The content assaying method of 12 kinds of compositions in a kind of Chinese medicinal composition preparation | |
| CN109374764B (en) | HPLC fingerprint spectrum and main component content determination method for eight-ingredient Longzui granules | |
| CN101028388B (en) | Quality inspection of Chinese-medicinal preparation for treating shortsighness and asthenopia | |
| CN104569252B (en) | A kind of method for building up of the finger printing of Chinese medicine composition | |
| CN106290599B (en) | Content determination method of traditional Chinese medicine composition | |
| CN103285306B (en) | Preparation method and detection method of traditional Chinese medicine composition for benefiting Qi and tonifying kidney | |
| CN109668970B (en) | Ultra-high performance liquid chromatography detection method for traditional Chinese medicine composition | |
| CN113466355A (en) | Construction method of high performance liquid phase characteristic spectrum of caulis sinomenii | |
| CN101966223A (en) | Fingerprint detection method for compound wintercreeper preparation | |
| CN113063885B (en) | Composition for preparing Baoyuan decoction, baoyuan decoction product and fingerprint spectrum measuring and quality detecting method thereof | |
| CN101513467A (en) | Method for controlling quality of dermatosis toxemia preparation | |
| CN100437112C (en) | Method for inspecting Chinese medicinal preparation quality in treatment of old man eyes dieases | |
| CN108459128B (en) | Quality control method of angelica sinensis Sini decoction composition | |
| CN101926889B (en) | Method for detecting white paeony root-medlar particles | |
| CN102293827B (en) | Quality detection method for camphor tree root and preparation containing camphor tree root | |
| CN108020613B (en) | Method for measuring content of menthol in traditional Chinese medicine composition | |
| CN109633006B (en) | Construction method and application of fingerprint of Weiyang ning | |
| CN109142563A (en) | A kind of construction method of guilingji capsules UPLC finger-print and its application | |
| CN101028474B (en) | Method for inspecting the quality of Chinese preparation with Yang-and kidney tonifying functions | |
| CN114942291A (en) | Method for detecting quality of 'Zhenyang Yangyin' granule | |
| CN107607665A (en) | One seedling medicine subprostrate sophora and its Chinese medicine preparation method of quality control |
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 |