CN113759045B - Characteristic spectrum of notopterygium root decoction pieces or formula granules, construction method of characteristic spectrum, notopterygium root formula granules, preparation method of notopterygium root formula granules and quality control method of notopterygium root formula granules - Google Patents
Characteristic spectrum of notopterygium root decoction pieces or formula granules, construction method of characteristic spectrum, notopterygium root formula granules, preparation method of notopterygium root formula granules and quality control method of notopterygium root formula granules Download PDFInfo
- Publication number
- CN113759045B CN113759045B CN202111070341.2A CN202111070341A CN113759045B CN 113759045 B CN113759045 B CN 113759045B CN 202111070341 A CN202111070341 A CN 202111070341A CN 113759045 B CN113759045 B CN 113759045B
- Authority
- CN
- China
- Prior art keywords
- peak
- solution
- notopterygium root
- notopterygium
- decursin
- 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
- G01N30/02—Column chromatography
- G01N30/50—Conditioning of the sorbent material or stationary liquid
- G01N30/52—Physical parameters
- G01N30/54—Temperature
-
- 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
- G01N30/62—Detectors specially adapted therefor
- G01N30/74—Optical detectors
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)
- Spectroscopy & Molecular Physics (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention belongs to the technical field of preparation of notopterygium root formula particles, and particularly relates to a characteristic spectrum of notopterygium root decoction pieces or formula particles and a construction method thereof, and a notopterygium root formula particles and a preparation method and a quality control method thereof. The method takes octadecylsilane chemically bonded silica as a filling agent, the mobile phase is acetonitrile-phosphoric acid solution, a specific elution program is adopted, 8 common characteristic peaks can be obtained under the elution condition, the effective separation of the common characteristic peaks is realized, the peak shape is good and free of interference, the quality of notopterygium incisum formula particles can be effectively judged, and the defect that the notopterygium incisum formula particles cannot be comprehensively, clearly and effectively detected due to interference caused by complex chemical components is overcome; in addition, in the construction method provided by the invention, the elution program time is short, and the construction time of the characteristic map is further shortened.
Description
Technical Field
The invention belongs to the technical field of preparation of notopterygium root formula particles, and particularly relates to a characteristic spectrum of notopterygium root decoction pieces or formula particles and a construction method thereof, and a notopterygium root formula particles and a preparation method and a quality control method thereof.
Background
Notopterygii Rhizoma et Radix Notopterygii, derived from dried rhizome and root of Notopterygium incisum Ting ex H.T. Chang or Notopterygium forbesii franchetii H.de Boiss. Collected in spring and autumn, removed fibrous root and silt, dried in the sun, the notopterygium root as a medicinal material is recorded in Shennong Ben Cao Jing (Shennong's herbal medicine), named after Qiang's land, has been used for thousands of years in China, is a common traditional Chinese medicine, and is mainly distributed in Qinghai, gansu, shaanxi, sichuan and Yunnan. Notopterygii rhizoma is pungent in flavor and warm in nature, has effects of relieving exterior syndrome, dispelling cold, dispelling pathogenic wind, removing dampness, and relieving pain, and is mainly used for treating wind-cold type common cold, headache, neck stiffness, rheumatism, paralysis, and soreness of shoulder and back.
Notopterygium incisum mainly contains chemical components such as volatile oil, coumarin, amino acid, organic acid, sterol and the like, and the pharmacological action and clinical application of the Notopterygium incisum are reflected in multiple aspects. Recent pharmacological studies show that notopterygium root medicinal materials have obvious effects of relieving pain and inflammation, improving blood circulation and enhancing the immunologic function of organisms, and the main active ingredients of the notopterygium root medicinal materials are coumarin compounds represented by notopterygium alcohol, decursin and isoimperatorin.
In recent years, with the continuous development of medical treatment and science and technology, a novel traditional Chinese medicine preparation, namely traditional Chinese medicine formula granules, which combines the unique traditional Chinese medicine of China with the modern science and technology appears, and the traditional Chinese medicine decoction pieces are prepared into the decoction-free granules, so that the decoction-free traditional Chinese medicine granule is convenient and simple for patients to take and is convenient to carry. Chinese patent document CN101474224A discloses a preparation method of notopterygium root formula granules, which adopts a straight-through steam method to extract volatile oil in decoction pieces, then the decoction pieces are decocted, and are subjected to reduced pressure concentration and then are included by beta-cyclodextrin, so that the process is more complicated, and individual heat-unstable components in the decoction pieces are degraded or changed in the process, so that the difference between the components of the formula granules and the components of the notopterygium root standard decoction is larger, and the quality of the notopterygium root formula granules is difficult to judge. In addition, in the prior art, when the quality of notopterygium root is detected, the separation degree of characteristic peaks in an obtained characteristic spectrum is poor, and the detection time is long.
Disclosure of Invention
Therefore, the technical problem to be solved by the invention is to overcome the defects that the quality of notopterygium root decoction pieces or formula granules is difficult to effectively judge, the separation degree of characteristic spectrums of preparations such as notopterygium root decoction pieces, formula granules and the like is poor and the like in the prior art, so that the characteristic spectrums of the notopterygium root decoction pieces or formula granules and the construction method thereof are provided.
Further, aiming at the problems that part of effective components are lost when preparing notopterygium root formula particles, the drug effect of the notopterygium root formula particles is difficult to keep consistent with that of traditional decoction, the process is complex and the like, the characteristic map of notopterygium root decoction pieces or formula particles and the construction method thereof, the notopterygium root formula particles and the preparation method and the quality control method thereof are provided.
Therefore, the invention provides the following technical scheme.
The invention provides a method for constructing a characteristic spectrum of notopterygium root decoction pieces or formula granules, which comprises the following steps,
(1) Preparing a test solution;
(2) Detecting the sample solution by high performance liquid chromatography, taking octadecylsilane chemically bonded silica as a filler, acetonitrile as a mobile phase A, and a solution containing phosphoric acid as a mobile phase B, and performing gradient elution, wherein the gradient elution procedure comprises the following steps: 0-6min, the volume percentage of the mobile phase A is 10-15%, and the volume percentage of the mobile phase B is 90-85%;6-9min, the volume percentage of the mobile phase A is 15-16%, and the volume percentage of the mobile phase B is 85-84%;9-14min, the volume percentage of the mobile phase A is 16-47%, and the volume percentage of the mobile phase B is 84-53%;14-18min, wherein the volume percent of the mobile phase A is 47-53%, and the volume percent of the mobile phase B is 53-47%;18-21min, the volume percentage of the mobile phase A is 53-90%, and the volume percentage of the mobile phase B is 47-10%.
The construction method also comprises a step of preparing a reference substance solution by adopting at least one of notopterygium alcohol, isoimperatorin, ferulic acid, chlorogenic acid, decursin and ferulic acid phenethyl alcohol ester, and a step of detecting the reference substance solution by high performance liquid chromatography in the construction method of claim 4 to obtain a reference substance characteristic map;
preferably, the preparation method of the control solution comprises the following steps: taking notopterygium alcohol, adding solvent, and making into notopterygium alcohol control solution A containing 10-20 μ g per 1 ml;
adding solvent into isoimperatorin to obtain isoimperatorin reference solution B containing 5-15 μ g of isoimperatorin per 1 ml;
adding solvent into ferulic acid, and making into ferulic acid control solution C containing 30-50 μ g per 1 ml;
adding solvent into chlorogenic acid, and making into chlorogenic acid reference solution D containing 50-70 μ g of chlorogenic acid per 1 ml;
taking decursin, adding solvent, and making into reference solution E containing decursin 150-250 μ g per 1 ml;
taking the ferulic acid phenethyl alcohol ester, adding a solvent to prepare a reference substance solution F containing 30-80 mu g of ferulic acid phenethyl alcohol ester per 1 ml;
more preferably, the solvent is selected from aqueous methanol, aqueous ethanol or water; the volume fraction of methanol in the methanol aqueous solution is 60-70%.
The preparation of the test solution comprises the following steps,
extracting Notopterygii rhizoma decoction pieces powder or Notopterygii rhizoma granule to obtain sample solution;
preferably, the construction method further comprises the step of extracting a notopterygium root reference medicinal material to obtain a notopterygium root reference medicinal material reference solution.
When preparing the notopterygium root decoction piece test sample solution, firstly, the notopterygium root decoction pieces are beaten into powder, then, methanol is added into the notopterygium root decoction piece powder, and the extraction and the filtration are carried out to obtain the notopterygium root decoction piece test sample solution; when preparing the notopterygium root formula particle test solution, adding methanol into the notopterygium root formula particle, extracting, and filtering to obtain the notopterygium root formula particle test solution.
The chromatographic conditions of the high performance liquid chromatography further comprise: the detection wavelength is 280-300nm; the flow rate is 0.2-0.6ml/min; the column temperature is 38-42 ℃; the sample amount is 1-5 muL;
preferably, 0.05-0.15% by volume of phosphoric acid in water is used as mobile phase B.
The invention provides a characteristic spectrum of notopterygium root formula granules obtained by the construction method.
The invention also provides a characteristic map of the notopterygium root formula particle, wherein the characteristic map of the notopterygium root formula particle is selected from any one of the following items (1) - (2);
(1) The characteristic map at least comprises 5 characteristic peaks, wherein the peak 2 is chlorogenic acid, the peak 3 is ferulic acid, the peak 5 is decursin, and the relative retention time of each characteristic peak is within +/-10% of a specified value;
taking peak No. 2 as a reference peak, and the relative retention time of peak No. 1 is 0.79;
taking the No. 3 peak as a reference peak, and the relative retention time of the No. 4 peak is 1.08;
(2) The characteristic map at least comprises 8 characteristic peaks, wherein the peak 2 is chlorogenic acid, the peak 3 is ferulic acid, the peak 5 is decursin, the peak 6 is notopterygium alcohol, the peak 7 is phenethyl ferulate, and the peak 8 is isoimperatorin; the relative retention time of each characteristic peak is within +/-10% of a specified value;
taking the No. 2 peak as a reference peak, and the relative retention time of the No. 1 peak is 0.79;
the relative retention time of peak No. 4 was 1.08 with peak No. 3 as the reference peak. .
In addition, the invention provides a preparation method of notopterygium root formula particles, which comprises the following steps,
(1) Processing Notopterygii rhizoma to obtain Notopterygii rhizoma decoction pieces;
(2) Decocting Notopterygii rhizoma decoction pieces to obtain Notopterygii rhizoma extractive solution, and concentrating to obtain fluid extract with relative density of 1.05-1.15 g/ml;
(3) Directly spray-drying the clear paste to obtain dry powder;
(4) And mixing the dry powder with auxiliary materials, and granulating to obtain the notopterygium root formula granules.
The inlet air temperature of the spray drying is 165-185 ℃.
The decocting method comprises adding water 8-12 times of decoction pieces into the first decoction, boiling for 30-60min, adding water 8-10 times of decoction pieces into the second decoction, boiling for 30-60min, and mixing filtrates to obtain Notopterygii rhizoma extract;
preferably, the cream yield of the notopterygium root extracting solution is 16-25%;
the processing steps comprise taking notopterygium root medicinal materials, spraying water, moistening for at least 12h, and drying at low temperature to obtain notopterygium root decoction pieces.
In the processing step, the specific step of moistening comprises the steps of taking notopterygium root medicinal materials, spraying water, turning over every 1 hour, observing whether the water is completely absorbed, and spraying the water again if the water is completely absorbed, wherein the spraying amount of the water is 10% of the amount of the medicinal materials, when the water adding amount is 40% of the amount of the medicinal materials, the moistening time is controlled to be at least 12 hours, the medicinal materials are basically moistened to reach the cutting standard, therefore, in the step of moistening the medicinal materials, the adding amount of the water is controlled to be 30-50% of the amount of the medicinal materials, and the moistening time is at least 12 hours.
In addition, the invention also provides notopterygium root formula particles prepared by the method.
Further, the invention provides a quality detection method of notopterygium root formula particles, which comprises the following steps,
(1) Preparing a reference substance solution E by adopting a decursin reference substance;
(2) Preparing a test solution;
(3) Performing high performance liquid chromatography, eluting with methanol and water as mobile phase at equal rate to obtain sample solution containing decursin 2-32mg;
preferably, the chromatographic conditions further comprise a detection wavelength of 330-340nm; the flow rate is 0.8-1.2ml/min; the column temperature is 28-32 ℃; the sample injection amount is 8-12 mu L;
more preferably, the control solution E contains 150-250 μ g of decursin per 1 ml.
The technical scheme of the invention has the following advantages:
1. the invention provides a method for constructing a characteristic spectrum of notopterygium root decoction pieces or formula particles, which takes octadecylsilane chemically bonded silica as a filling agent, takes acetonitrile-phosphoric acid solution as a mobile phase, adopts a specific elution program, can obtain 8 common characteristic peaks under the elution condition, realizes effective separation of the common characteristic peaks, has good peak shape without interference, can effectively judge the quality of the notopterygium root formula particles, and overcomes the defect that the quality of the notopterygium root formula particles cannot be comprehensively, clearly and effectively detected due to interference caused by complex chemical components; in addition, in the construction method provided by the invention, the elution procedure time is short, so that the construction time of the characteristic spectrum is shortened, and meanwhile, the method also has the advantages of good stability, high precision and good repeatability. By controlling the elution gradient, chlorogenic acid, ferulic acid, notopterygium alcohol and isoimperatorin can be effectively separated, and the reliability of a characteristic map is improved.
The construction method of the characteristic spectrum of the notopterygium root formula particle provided by the invention is also suitable for performing quality control on notopterygium root decoction pieces.
2. According to the method for constructing the characteristic map of the notopterygium root medicinal material, the decoction pieces or the formula granules, the characteristic map constructed by the method identifies at least one of four effective components of chlorogenic acid, ferulic acid, notopterygium root alcohol and isoimperatorin, so that the characteristic map is more accurate, stable and reliable, and the reliability of the characteristic map is improved.
3. According to the preparation method of the notopterygium root formula particle, auxiliary materials are not required to be added in the spray drying process, the content of the effective components of the midbody of the notopterygium root formula particle can be ensured, the problems that the effective components are degraded in the extraction process to cause the loss of the effective components and the like are solved, meanwhile, the obtained spray drying powder is fine and smooth in powder quality, good in dissolubility and fluidity and easy to collect, the powder yield at least reaches 85%, and the drying time and the using amount of the auxiliary materials are greatly saved.
The notopterygium root formula particle provided by the invention can keep consistent with the effective components of a notopterygium root standard decoction, so that the drug effect of the formula particle is ensured; compared with the prior art, the volatile oil in the notopterygium root decoction pieces is not required to be extracted and included when the notopterygium root formula particles are prepared, the preparation process flow is simplified, the energy is saved, the efficiency is high, the stability is good, and the preparation method is suitable for large-scale industrial production.
By controlling the extraction process parameters of the notopterygium root formula particles, the invention can effectively ensure that the effective components of decursin, chlorogenic acid, ferulic acid and the like in the notopterygium root formula particles are fully extracted, and reduce the loss of the effective components.
4. According to the quality detection method of the notopterygium root formula particles, the quality of the notopterygium root formula particles is detected according to the content of decursin, so that the quality of the notopterygium root formula particles can be effectively evaluated.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without creative efforts.
FIG. 1 is a characteristic spectrum of lyophilized powder obtained from each batch of decoction pieces in example 1 of the present invention;
FIG. 2 is a characteristic spectrum of a Notopterygium incisum reference drug substance in example 7 of the present invention;
FIG. 3 is a characteristic spectrum of a control in example 7 of the present invention;
FIG. 4 is a characteristic diagram of a sample solution obtained by extracting different solvents in section 1.1 of Experimental example 1 of the present invention;
FIG. 5 is a characteristic map of a test solution in section 2.1 of Experimental example 1 of the present invention;
FIG. 6 is a characteristic map of a test solution in section 2.2 of Experimental example 1 of the present invention;
FIG. 7 is a characteristic diagram of a sample solution under different column temperatures in section 2.3 of Experimental example 1 of the present invention;
FIG. 8 is a characteristic diagram of the sample solution at different flow rates in section 2.4 of Experimental example 1 of the present invention;
FIG. 9 is a characteristic diagram of a sample solution of the present invention in Experimental example 1, at different concentrations of mobile phase B in section 2.5;
FIG. 10 is a characteristic diagram of the sample solution under different chromatographic columns of section 2.6 in Experimental example 1 of the present invention;
FIG. 11 is a characteristic map of a decursin reference solution in section 2.1 of Experimental example 2 of the present invention;
FIG. 12 is a characteristic diagram of the Notopterygium incisum sample solution in section 2.1 of Experimental example 2 of the present invention;
FIG. 13 is a characteristic diagram of a sample solution at different column temperatures in section 2.3 of Experimental example 2 of the present invention;
FIG. 14 is a characteristic diagram of a sample solution of section 2.4 at different flow rates in Experimental example 2 of the present invention;
FIG. 15 is a graph of the regression equation of section 3.5 in Experimental example 2 of the present invention;
FIG. 16 is a characteristic diagram of a test solution of Notopterygium incisum decoction pieces in example 9 of the present invention.
Detailed Description
The following examples are provided to better understand the present invention, not to limit the best mode, and not to limit the content and protection scope of the present invention, and any product that is the same or similar to the present invention and is obtained by combining the present invention with other features of the prior art and the present invention falls within the protection scope of the present invention.
The examples do not show the specific experimental steps or conditions, and can be performed according to the conventional experimental steps described in the literature in the field. The reagents or instruments used are conventional reagent products which are commercially available, and manufacturers are not indicated.
Example 1
The present embodiment considers the cream yield, and specifically includes the following steps,
taking 100g of each of 16 batches of notopterygium root decoction pieces (see table 1 in batches and manufacturers), adding 7 times of water, soaking for 30min, heating to boil, decocting for 20min, filtering while hot by 150-mesh filter cloth, adding 5 times of water into decoction dregs, heating to boil, decocting for 15min, filtering while hot by 150-mesh filter cloth, combining the two filtrates to obtain notopterygium root extract, cooling to room temperature in water bath, and weighing the total mass of the notopterygium root extract. Weighing 1/10 of the total mass of Notopterygii rhizoma extract, and placing in an evaporation pan, and recording as M 1 Drying in water bath, drying at 105 deg.C for 5 hr, cooling for 30min, repeating the steps until constant weight is reached, and recording as M 2 Calculating the cream yield according to the formula 1;
wherein M is 1 Represents 1/10 of the total mass of the notopterygium root extract, and the unit is g; m is a group of 2 The total mass of the dried liquid medicine and the evaporating dish is represented as g; m is a group of 0 Represents the mass of the evaporating dish in g; n represents the total mass of the liquid medicine, and the unit is g; w represents the mass of notopterygium root decoction pieces in g.
Concentrating the residual extractive solution at 65 deg.C under reduced pressure until the ratio of fluid extract to decoction pieces is about 1, transferring to freeze drying tray, vacuum freeze drying (specifically comprises drying at-50 deg.C for 4 hr, drying at-35 deg.C for 4 hr, drying at-30 deg.C for 6 hr, drying at-25 deg.C for 6 hr, drying at-20 deg.C for 3 hr, drying at-15 deg.C for 3h, drying at 0 deg.C for 3 hr, drying at 15 deg.C for 3 hr, drying at 35 deg.C for 8 hr, and vacuum degree of 10 Pa), collecting lyophilized powder, making into sample solution, and measuring content of decursin in lyophilized powder. Meanwhile, the notopterygium root decoction pieces are ground into powder to be used as a test solution for measuring the content of decursin in the decoction pieces. The determination method of the content of decursin comprises the following steps:
preparation of a test solution: taking 0.2g of a test sample (freeze-dried powder or notopterygium root decoction piece powder), accurately weighing, placing in a conical flask with a plug, accurately adding 20ml of 70% methanol, weighing, carrying out ultrasonic treatment (power 250W and frequency 40 kHZ) for 30min, cooling, weighing again, complementing the weight loss with 70% methanol, shaking up, filtering, and taking the obtained filtrate as a test sample solution.
Preparation of control solutions: taking a proper amount of decursin reference substance, precisely weighing, and adding methanol to obtain reference substance solution E containing 200 μ g per 1 ml.
According to high performance liquid chromatography, octadecylsilane chemically bonded silica is used as a filler, methanol-water (40. Accurately sucking 10 μ l of the decursin reference solution E and the test solution respectively, injecting into a liquid chromatograph, and measuring to obtain content of decursin in lyophilized powder and decoction pieces according to the standard 0512 of the 2020 version of Chinese pharmacopoeia.
The content (mg/g) of decursin in the lyophilized powder prepared from the decoction pieces of 16 batches and the content (mg/g) of decursin in the decoction pieces of notopterygium root of 16 batches were calculated according to the formula 2, and the results are shown in table 1.
TABLE 1 extraction rate of Notopterygii rhizoma extract, notopterygii rhizoma lyophilized powder and herba Philippici glycoside content of decoction pieces
According to the above experiment results, the average cream yield of 16 batches is 20.69%, and + -20% is taken as the cream yield range of the notopterygium root extract, namely 16-25%, and when the average cream yield exceeds the range, the quality of the notopterygium root decoction pieces is unqualified.
Determining lyophilized powder obtained from each batch of Notopterygii rhizoma decoction pieces by high performance liquid chromatography to obtain characteristic chromatogram of each batch, which comprises:
preparation of a test solution: weighing Notopterygii rhizoma lyophilized powder 0.2g, precisely weighing, placing in conical flask with stopper, precisely adding 70% methanol 20ml, weighing, ultrasonic treating (power 250W, frequency 40 kHZ) for 30min, cooling, weighing again, supplementing lost weight with 70% methanol, shaking, filtering, and collecting filtrate as sample solution.
Chromatographic conditions and system applicability test: octadecylsilane chemically bonded silica is used as filler (Waters ACUITY)
BEH C18.1X 100mm,1.7 μm) chromatography column; acetonitrile is used as a mobile phase A, a phosphoric acid aqueous solution with the volume fraction of 0.1% is used as a mobile phase B, and the time is 0-6min, wherein the volume percentage of the mobile phase A is 10-15%, and the volume percentage of the mobile phase B is 90-85%;6-9min, wherein the volume percent of the mobile phase A is 15-16%, and the volume percent of the mobile phase B is 85-84%;9-14min, the volume percentage of the mobile phase A is 16-47%, and the volume percentage of the mobile phase B is 84-53%;14-18min, wherein the volume percent of the mobile phase A is 47-53%, and the volume percent of the mobile phase B is 53-47%;18-21min, wherein the volume percent of the mobile phase A is 53-90%, and the volume percent of the mobile phase B is 47-10%;21-23min, wherein the volume percent of the mobile phase A is 90-10%, and the volume percent of the mobile phase B is 10-90%; the flow rate was 0.4ml per minute; the detection wavelength is 290nm; the column temperature was 40 ℃.
The characteristic spectrum of lyophilized powder obtained from each batch is shown in figure 1, S1-S16 in figure 1 is compared with characteristic spectrum of lyophilized powder obtained from decoction pieces of each batch of YP-QH-1 to YP-QH-16, and R is standard decoction of Notopterygii rhizoma. The chromatogram corresponding to each batch of notopterygium root freeze-dried powder is analyzed by software of a traditional Chinese medicine chromatogram fingerprint similarity evaluation system (2012 edition), the time window is set to be 0.1, 16 batches of sample superposition spectrums and comparison characteristic spectrums thereof are obtained by automatic matching after multipoint correction by adopting an average number method, and at least 5 common peaks are determined. The similarity evaluation results are shown in table 1, and the similarity value of the 16 batches of notopterygium root freeze-dried powder test samples is in the range of 0.953-0.998.
Example 2
The content of volatile oil in notopterygium root decoction pieces is examined in the embodiment, specifically as follows,
the volatile oil is one of the main drug effect components of notopterygium root, and the content of the volatile oil is taken as one of the quality control indexes in Chinese pharmacopoeia, so that the content of the volatile oil in notopterygium root decoction pieces and notopterygium root standard decoction is measured by taking the extraction rate of the volatile oil as an evaluation index. The method comprises the following specific steps:
respectively placing 16 batches of notopterygium root decoction pieces in a round-bottom flask, adding 500ml of water and a plurality of glass beads, shaking and mixing, connecting a volatile oil tester and a reflux condenser tube, heating and refluxing to extract volatile oil, distilling and extracting the volatile oil by the extraction method according to a determination method (A method) of the volatile oil in appendix of China pharmacopoeia of 2020 edition, and calculating the extraction rate of the volatile oil according to a formula 3.
Detecting volatile oil content in Notopterygii rhizoma standard decoction, respectively taking 16 batches of Notopterygii rhizoma decoction pieces each 100g, soaking in 7 times of water for 30min, heating to boil, decocting for 20min, filtering with 150 mesh filter cloth while it is hot, adding 5 times of water into residue, heating to boil, decocting for 15min, filtering with 150 mesh filter cloth while it is hot, mixing the filtrates, and cooling Notopterygii rhizoma extract to room temperature in water bath to obtain Notopterygii rhizoma standard decoction; then respectively placing the Notopterygii rhizoma standard decoction in a round-bottom flask, adding several glass beads, connecting with volatile oil detector and reflux condenser tube, and heating to extract volatile oil by the same method; the contents of volatile oil in the notopterygium root decoction pieces and the notopterygium root standard decoction are shown in table 2.
TABLE 2 content of volatile oil in Notopterygii rhizoma decoction pieces and Notopterygii rhizoma standard decoction
| Batch number | Extraction rate (%) of volatile oil in decoction pieces | Extraction rate (%) of volatile oil in standard decoction |
| YP-QH-01 | 2.2 | -- |
| YP-QH-02 | 4.0 | -- |
| YP-QH-03 | 2.0 | -- |
| YP-QH-04 | 2.2 | -- |
| YP-QH-05 | 2.6 | -- |
| YP-QH-06 | 2.5 | -- |
| YP-QH-07 | 1.6 | -- |
| YP-QH-08 | 1.4 | -- |
| YP-QH-09 | 1.6 | -- |
| YP-QH-10 | 1.8 | -- |
| YP-QH-11 | 2.7 | -- |
| YP-QH-12 | 1.6 | -- |
| YP-QH-13 | 1.8 | -- |
| YP-QH-14 | 3.0 | -- |
| YP-QH-15 | 2.6 | -- |
| YP-QH-16 | 2.8 | -- |
Note: in the table, "- -" indicates no reading and the volatile oil content is too low.
The experimental results in table 2 show that the volatile oil content in the notopterygium root standard decoction is low, and the volatile oil content in notopterygium root decoction pieces conforms to the range (not less than 1.4%) specified in 2020 version of Chinese pharmacopoeia, so that the volatile oil does not need to be added when preparing notopterygium root formula particles, the step of extracting the volatile oil and then performing inclusion when preparing the notopterygium root formula particles in the prior art is overcome, the process flow is simplified, the cost is reduced, and the production efficiency is improved while the drug effect consistency of the notopterygium root formula particles and the notopterygium root standard decoction is ensured.
Example 3
This example examined the effect of concentration temperature on the content of decursin in the test solution
Taking 200g of Notopterygii rhizoma decoction pieces (YP-QH-14), decocting with 7 times of water, soaking for 30min, heating to boil, and decocting for 60min; adding water 5 times the amount of decoction pieces, heating to boil, decocting for 40min, filtering the extractive solution with 150 mesh filter cloth, and mixing filtrates; the liquid medicine is cooled to room temperature in water bath, and the total mass of the liquid medicine is weighed. Dividing the liquid medicine into 32 equal parts, respectively placing the 32 equal parts in a centrifuge tube, weighing and sealing, placing 2 parts in room temperature, preparing the liquid medicine into a test solution for detection, placing the remaining 30 parts in a water bath heating condition at the temperature of 65 ℃, 75 ℃ and 85 ℃, placing 10 parts in each temperature, heating to 3 hours, 6 hours, 9 hours, 15 hours and 24 hours, respectively taking 2 parts (parallel samples) in each temperature condition, then placing the samples in the room temperature, supplementing the weight, and obtaining 32 parts of notopterygium root concentrated temperature investigation samples in total. Preparing a test solution according to a content determination method under the quality standard item of the notopterygium root intermediate, detecting, taking an average value of the same sample, and obtaining a test result shown in table 3.
TABLE 3 concentration temperature of Notopterygium incisum to examine the content (mg/g) of decursin in the sample
Note: the results in the table show that the content of decursin in each gram of notopterygium root decoction pieces is mg/g.
From the results in table 3, it can be seen that the content of decursin is less affected by the temperature, and when the concentration temperature is 65 ℃ and 75 ℃, the content of decursin is substantially stable with the increase of time and temperature; when the concentration temperature is 85 deg.C, the content of decursin is reduced after the temperature is maintained for more than 15 hr. Therefore, the concentration temperature of the notopterygium root extract is controlled below 75 ℃, and the concentration time is controlled within 24 hours.
Example 4
This example investigated the effect of different extraction process parameters on the cream yield and the nodakenin content
Performing L with the paste yield and the content of decursin as indexes by orthogonal experiment method 9 (3 4 ) And (4) performing orthogonal experiment, investigating water addition quantity, extraction times and extraction time which influence the extraction effect, and determining the optimal extraction process condition. The factor levels are shown in Table 4, the experimental protocol is shown in Table 5, and the experimental results are shown in Table 6.
TABLE 4 orthogonal experiment factor horizon
Note: the water addition in the table refers to the water addition (A) of the first decoction, the extraction frequency is the decoction frequency, the water addition of the second decoction is A-2 times, and the water addition of the third decoction is A-2 times.
TABLE 5 orthogonal Experimental protocols
The method comprises the following specific steps: taking 9 parts of notopterygium root decoction pieces (with the batch number of YP-QH-14), placing 100g of notopterygium root decoction pieces in a round bottom flask, extracting according to the method in the table 5, filtering an extracting solution through 150-mesh filter cloth, combining filtrates, cooling to room temperature in a water bath, weighing the total mass of the liquid medicine, determining the paste yield according to the method in the example 1, concentrating the residual extracting solution at 65 ℃ under reduced pressure until the ratio of the clear paste to the decoction pieces is about 1, transferring to a freeze-drying tray, carrying out vacuum freeze drying (the method is shown in the example 1), collecting freeze-dried powder, detecting the content of decursin in the freeze-dried powder according to the method in the example 1, and calculating the decursin transfer rate.
TABLE 6 orthogonal experimental results table
| Test No. | Percentage of cream discharged (%) | Philippine violet root picroside content (mg/g) in freeze-dried powder | Transfer Rate (%) |
| 1 | 14.6 | 39.8 | 49.5 |
| 2 | 23.9 | 37.0 | 75.5 |
| 3 | 27.8 | 37.1 | 87.8 |
| 4 | 18.0 | 37.8 | 58.0 |
| 5 | 25.9 | 38.5 | 85.0 |
| 6 | 27.5 | 42.1 | 98.6 |
| 7 | 21.0 | 39.3 | 70.3 |
| 8 | 24.9 | 39.3 | 83.4 |
| 9 | 28.5 | 35.3 | 85.9 |
Note: the content of decursin in 1g decoction pieces is 11.7mg.
Inputting the results of the paste yield and the decursin transfer rate of the orthogonal experiment into Design-Expert10.0.4.0 for processing, comprehensively evaluating the results of the notopterygium root standard decoction by taking factors such as losses in the comprehensive production process as target values, and preferably selecting the extraction process composition closest to the standard decoction as follows: adding water 8-12 times of decoction pieces into the first decoction, boiling and extracting for 30-60min, adding water 8-10 times of decoction pieces into the second decoction, boiling and extracting for 30-60min, and mixing filtrates to obtain Notopterygii rhizoma extract; preferably, water with the amount 11 times of the decoction pieces is added into the first decoction, and boiling extraction is carried out for 40min; decocting with 9 times of water, boiling and extracting for 40min.
Example 5
This example examined the influence of the temperature of the inlet air and the type of the auxiliary materials on the yield and the content of decursin
Performing L with the paste yield and the content of decursin as indexes by orthogonal experiment method 9 (3 4 ) And (4) performing orthogonal experiments, inspecting the relative density of the clear paste, the air inlet temperature, the additive amount of the auxiliary materials and the types of the auxiliary materials, and determining the optimal extraction process conditions. The factor levels are shown in Table 7, the experimental protocol is shown in Table 8, and the experimental results are shown in Table 9. In table 7, the ratio of the auxiliary materials is the percentage of the auxiliary materials in the amount of the decoction pieces.
TABLE 7 orthogonal experiment factor horizon
Table 8 orthogonal experimental protocol table
The method comprises the following specific steps: taking 2000g of Notopterygii rhizoma decoction pieces (batch number YP-QH-14), adding water 7 times of the decoction pieces in the first decoction, soaking for 30min, heating to boil, and decocting for 20min; adding water 5 times the amount of the decoction pieces, boiling, decocting for 15min, filtering the extractive solution with 150 mesh filter cloth, mixing filtrates, concentrating under reduced pressure at 65 deg.C to density set on the table, and performing experiment according to the parameters set in table 8. And after drying, collecting spray-dried powder, weighing the powder, calculating the powder yield according to a formula 4, and sampling to determine the water content, the dissolution rate and the content of decursin in the dried powder. The method for testing the moisture content of the dry powder comprises the following steps of weighing part of the dry powder, placing the weighed dry powder in a drying oven to be dried to constant weight, and calculating the moisture content of the dry powder; the melting rate was measured by taking a sample of 0.5g and precisely weighing W 1 Is arranged atDried in a 50ml centrifuge tube of constant weight, weighed as W 0 Adding hot water 10ml precisely, stirring and shaking for 5min, centrifuging at 3000r/min for 15min, discarding supernatant, oven drying residue at 80 deg.C to constant weight (the difference between the two previous weighing is less than 5 mg), precisely weighing W 2 The melting rate was calculated by referring to equation 5.
TABLE 9 orthogonal Experimental results Table
Note: the blowing frequency is 35Hz; peristaltic pump speed 15rpm.
The experimental results in table 9 show that the experimental groups 1, 5 and 9 without the auxiliary materials all can obtain higher powder yield, which is more than 85%, and the powder is slightly sticky to the wall in the spraying process, dry and loose, and easy to collect. The spray-dried powder obtained in experiments 1, 5 and 9 has moderate water content, high dissolution rate, and insignificant difference of effective component content and slightly higher than the effective component content in the standard decoction, thereby ensuring the quality of the intermediate of the notopterygium root formula granule. Thus, the parameters chosen are as follows: the density of the material is 1.05-1.15g/ml (measured at 60 ℃); the air inlet temperature is 165-185 ℃, and no auxiliary materials are added in the spray drying process.
Example 6
This example provides a method for preparing notopterygium root formula particles, which comprises the following steps,
(1) Respectively taking 9kg of notopterygium root medicinal materials (with the batch numbers of YP-QH-14, YP-QH-15 and YP-QH-16 respectively), removing impurities, cleaning silt, spraying water for 4 times, wherein the spraying amount of water is 10% of the amount of the notopterygium root medicinal materials, the interval time between every two adjacent water spraying is 1h, then moistening for 12h, thoroughly permeating the medicinal materials (the medicinal materials are soaked thoroughly, the section has no white core, and no water remains), cutting into thick pieces (the thickness is 2-4 mm), and drying at the temperature lower than 50 ℃ to obtain the notopterygium root medicinal materials.
(2) Placing the above Notopterygii rhizoma decoction pieces in an extraction pot, adding 11 times of water into the first decoction, heating to boil, extracting for 40min, adding 9 times of water into the second decoction, heating to boil, extracting for 40min, filtering the extractive solution with 150 mesh filter cloth, and mixing the filtrates to obtain Notopterygii rhizoma extract; then concentrating the Notopterygii rhizoma extract at 75 deg.C to obtain fluid extract with relative density of 1.05-115g/ml (measured at 60 deg.C).
(3) Heating the above fluid extract to 60 deg.C, and spray drying directly to obtain dry powder, wherein the air inlet temperature of spray drying is 175 deg.C, the frequency of atomizer is 60Hz, and the feeding speed is 24Hz.
(4) Adding appropriate amount of maltodextrin into Notopterygii rhizoma dried powder, mixing, and controlling the compression roller gap of granulator to 0.6-2.5mm; the rotating speed of the compression roller is 30rpm; and (4) granulating under the pressure of 80-100bar by using a 16-mesh sieve and a 60-mesh sieve, and packaging with a medicinal composite membrane to obtain the notopterygium root formula granules with the batch numbers of KL-QH-14, KL-QH-15 and KL-QH-16.
Example 7
The embodiment provides a method for constructing a characteristic map of notopterygium root formula particles, which comprises the following steps,
(1) Preparation of control solutions:
taking a proper amount of notopterygium alcohol reference substance, precisely weighing, and respectively adding 70% methanol to obtain 15 μ g of notopterygium alcohol reference substance solution A per 1 ml;
precisely weighing an appropriate amount of isoimperatorin reference substance, and adding 70% methanol to obtain 5 μ g of isoimperatorin reference substance solution B per 1 ml;
taking a proper amount of ferulic acid reference substance, precisely weighing, and adding 70% methanol to prepare ferulic acid reference substance solution C containing 40 μ g per 1 ml;
precisely weighing appropriate amount of chlorogenic acid reference substance, and adding 70% methanol to obtain chlorogenic acid reference substance solution D containing 60 μ g of chlorogenic acid per 1 ml;
taking a proper amount of decursin reference substance, precisely weighing, and respectively adding 70% methanol to prepare decursin reference substance solution E containing 200 μ g of decursin per 1 ml;
taking a proper amount of the reference substance of the ferulic acid phenethyl alcohol ester, precisely weighing, and adding 70% methanol to obtain a reference substance solution F containing 50 mu g of the ferulic acid phenethyl alcohol ester per 1 ml.
(2) Preparation of reference solution of reference drug: weighing 1g of Notopterygii rhizoma reference medicinal material (batch No. 121405-200401), precisely weighing, placing in a conical flask with a plug, adding 30ml of water, decocting for 30min, filtering, evaporating to dryness in water bath, precisely adding 20ml of 70% methanol, ultrasonic treating (power 250W, frequency 40 kHZ) for 30min, shaking, filtering, and collecting the filtrate as Notopterygii rhizoma reference medicinal material reference solution.
(3) Preparation of a test solution: taking 0.2g of each of three batches of notopterygium root formula particles (batch numbers: KL-QH-14, KL-QH-15 and KL-QH-16), precisely weighing, placing in a conical flask with a stopper, precisely adding 20ml of 70% methanol, weighing, ultrasonically treating (power 250W and frequency 40 kHZ) for 30min, cooling, weighing again, complementing the lost weight with 70% methanol, shaking up, filtering, and taking the obtained filtrate as a test solution.
(4) Precisely absorbing 2 μ L of each of the test solution, notopterygii rhizoma reference solution and reference solution, injecting into high performance liquid chromatograph, and measuring under chromatographic conditions: octadecylsilane chemically bonded silica (Waters ACQUITY UPLC BEH C18, 2.1X 100mm,1.7 μm) as filler, acetonitrile as mobile phase A, and phosphoric acid aqueous solution with volume fraction of 0.1% as mobile phase B, and eluting according to the following proportion gradient: 0-6min, the volume percentage of the mobile phase A is 10-15%, and the volume percentage of the mobile phase B is 90-85%;6-9min, the volume percentage of the mobile phase A is 15-16%, and the volume percentage of the mobile phase B is 85-84%;9-14min, the volume percentage of the mobile phase A is 16-47%, and the volume percentage of the mobile phase B is 84-53%;14-18min, wherein the volume percent of the mobile phase A is 47-53%, and the volume percent of the mobile phase B is 53-47%;18-21min, wherein the volume percent of the mobile phase A is 53-90%, and the volume percent of the mobile phase B is 47-10%; (ii) a The detection wavelength is 290nm; the flow rate is 0.4ml/min, the column temperature is 40 deg.C, and characteristic patterns of Notopterygii rhizoma granule sample solution, notopterygii rhizoma reference medicinal material solution and reference solution are respectively obtained, wherein the reference solution characteristic pattern of Notopterygii rhizoma reference medicinal material is shown in figure 2, and the reference solution characteristic pattern is shown in figure 3.
According to the principle of good stability, separation degree and peak shape of relative retention time, 8 common characteristic peaks are selected as reference substance solution of notopterygium root reference medicinal materials, wherein the peak 2 is chlorogenic acid, the peak 3 is ferulic acid, the peak 5 is decursin, the peak 6 is notopterygium alcohol, the peak 7 is phenethyl ferulate, and the peak 8 is isoimperatorin; taking the No. 2 peak and the No. 3 peak as reference peaks, wherein the relative retention time of each characteristic peak is within +/-10% of a specified value; taking peak No. 2 as a reference peak, and the relative retention time of peak No. 1 is 0.79; the relative retention time of peak No. 4 was 1.08 with peak No. 3 as the reference peak.
Comparing the characteristic maps of three batches of Notopterygii rhizoma formulation granule test solution (KL-QH-14, KL-QH-15, KL-QH-16) and Notopterygii rhizoma reference medicinal material reference solution, taking peak No. 2 as reference peak, and keeping the relative retention time of peak No. 1 at 0.79; the relative retention time of peak No. 4 was 1.08 with peak No. 3 as the reference peak, and was within ± 10% of the specified value.
Experimental example 1
1. Examination of preparation of test solution
1.1 selection of extraction solvent
The extraction solvent is water, methanol, 70% methanol, 50% methanol, 30% methanol, ethanol, 70% ethanol, 50% ethanol, and 30% ethanol. The preparation method of the test solution comprises the following steps: grinding a plurality of same test samples (KL-QH-14), precisely weighing about 0.2g, placing in a conical flask with a plug, adding 20ml of extraction solvent, sealing the plug, weighing, carrying out ultrasonic treatment for 30min (power 250W and frequency 40 kHz), taking out, cooling, weighing again, adding the extraction solvent to complement weight loss, shaking up, and filtering to obtain a test sample solution. Precisely sucking 2 μ L of the sample solution, injecting into an ultra high performance liquid chromatograph, and measuring according to the chromatographic conditions of the example 7, wherein the results are shown in Table 10 and FIG. 4;
TABLE 10 characteristic profiles of test solutions obtained with different extraction solvents
From the above results, it is understood that when the sample solution is prepared by using water, methanol and ethanol as the extraction solvent, the extraction ability is weak and the extraction is incomplete, that when 70% ethanol and 50% ethanol are used as the extraction solvent, the whole peak shape of the characteristic spectrum of the sample solution is poor, that when 30% ethanol is used as the extraction solvent, the peak shape of the No. 2 peak is poor, and that when 70% methanol, 50% methanol and 30% methanol are used as the extraction solvent, the peak shape of the characteristic spectrum is good. Considering the factors of pattern peak shape, convenient operation and the like, 70 percent methanol is preferably used as an extraction solvent.
1.2 selection of extraction method
Taking an extraction mode as a variable, respectively extracting and preparing a sample solution by adopting two methods of ultrasonic extraction and heating reflux extraction, wherein the specific method comprises the following steps: taking a proper amount of the same test sample (KL-QH-14), grinding, taking 0.2g, precisely weighing, placing in a conical flask with a plug, precisely adding 20ml of 70% methanol, sealing the plug, weighing, respectively heating and refluxing (85 ℃, heating in a water bath) and treating with ultrasound (power 250W, frequency 40 kHZ) for 30min, taking out, cooling, complementing the lost weight, shaking uniformly, filtering, taking a subsequent filtrate, and determining the result according to the method of example 7, wherein the result is shown in Table 11.
TABLE 11 characteristic maps of test solutions obtained by different extraction methods
From the above results, the peak area difference of each characteristic peak in the characteristic spectrum obtained by using the two extraction methods of ultrasound and reflux is not large, and ultrasound extraction is preferred in consideration of convenience of ultrasound extraction operation.
1.3 selection of the amount of extraction solvent added
With the addition amount of the extraction solvent as a variable, 10ml, 20ml and 50ml of the extraction solvent were added, and a sample solution was prepared according to section 1.1 in this experimental example 1, and a characteristic spectrum was obtained by measuring according to the chromatographic conditions of example 7, and the results are shown in table 12.
TABLE 12 characteristic spectra of test solutions obtained with different amounts of extraction solvent
From the above results, it can be seen that after the same solvent amount is converted, the peak area difference of each solvent amount of the three groups is not large, which indicates that the three groups are completely extracted, and considering the factors of moderate height, solvent saving, etc., the solvent extraction amount is 20ml as the optimal addition amount, i.e., 20ml of 70% methanol is added for each 0.2g of the sample.
1.4 examination of extraction time
With the ultrasonic extraction time as a variable, the ultrasonic extraction time is 20min, 30min and 40min respectively, the test sample solution prepared according to the method in section 1.1 of the experimental example is measured according to the chromatographic conditions of the example 7, and the results are shown in table 13.
TABLE 13 characteristic spectra of test solutions obtained at different extraction times
From the above results, it was found that the peak areas of the characteristic peaks obtained at different extraction times were not very different, and that the area of peak 1 at 20min was slightly lower, and the extraction time selected was preferably 30min for more complete extraction.
2. Selection of chromatographic conditions
2.1 selection of detection wavelength
The test solution was prepared according to the method of example 7, and the notopterygium root test solution was subjected to 3D full scan, showing that the peak around 290nm was abundant and the peak shape was better than other wavelengths. 290nm is therefore preferred as the detection wavelength, and the profile is shown in FIG. 5.
2.2 delay test
A test solution was prepared according to the method of example 7 and measured with reference to the chromatographic conditions of example 7, and the results of 2-fold elution time of the mobile phase were recorded, as shown in FIG. 6, indicating that no other chromatographic peak was present after 21 min.
2.3 selection of column temperature
Taking the column temperatures as variables, the column temperatures are respectively 38 ℃, 40 ℃ and 42 ℃, and the rest is the same as example 7, taking the same notopterygium root sample solution, measuring at different column temperatures to obtain a characteristic map, as shown in figure 7, the relative retention times of No. 1 peak, no. 2 peak, no. 3 peak and No. 4 peak are shown in Table 14.
TABLE 14 relative retention times and float ranges of characteristic peaks at different column temperatures
From the above results, it can be seen that the relative retention times of the characteristic peaks at different column temperatures are within the range of + -10% of the specified values, indicating that the method has good durability for different column temperatures.
2.4 selection of flow Rate
Taking the flow rates as variables, the flow rates are respectively 0.38ml/min, 0.40ml/min and 0.42ml/min, the rest is the same as the example 7, the same notopterygium root sample solution is taken and measured under different flow rates to obtain a characteristic spectrum, the relative retention time of the No. 1 peak, the No. 2 peak, the No. 3 peak and the No. 4 peak is shown in the table 15.
TABLE 15 relative retention times and float ranges of characteristic peaks at different flow rates
From the above results, it can be seen that the relative retention times of the characteristic peaks at different flow rates are within ± 10% of the specified values, indicating that the method is robust to different flow rates.
2.5 selection of the concentration of the Mobile phase
Taking the concentration of the mobile phase B (phosphoric acid aqueous solution) as a variable, the volume fractions of the mobile phase B are respectively 0.08%, 0.10% and 0.12%, and the rest is the same as example 7, the same notopterygium root sample solution is taken and measured under different mobile phase B concentrations to obtain a characteristic spectrum, which is shown in Table 16 as the relative retention time of the No. 1 peak, the No. 2 peak, the No. 3 peak and the No. 4 peak.
TABLE 16 relative retention times and float ranges of characteristic peaks at different mobile phase B concentrations
From the above results, it can be seen that the relative retention times of the characteristic peaks at different acid concentrations were within the specified range of ± 5%, indicating that the method is more robust to different mobile phase B concentrations.
2.6 selection of chromatography columns
Taking chromatographic columns as variables, and taking the same notopterygium root test sample solution as in example 7 for the rest, and measuring under different chromatographic columns to obtain characteristic maps, wherein each chromatographic column is as follows as shown in figure 10; the relative retention times of peak 1, peak 2, peak 3, and peak 4 are shown in Table 17.
A chromatographic column 1: waters ACUITY
BEH C18(2.1×100mm，1.7μm)
And (3) chromatographic column 2: waters
T3(2.1×100mm，1.6μm)
A chromatographic column 3: waters ACUITY
HSS T3(2.1×100mm，1.8μm)
TABLE 17 investigation of relative retention times and fluctuation ranges of characteristic peaks for different chromatographic columns
From the above results, it can be seen that the chromatogram peak times of different chromatographic columns are slightly different, but the relative retention times are within ± 10% of the specified values, when the chromatographic column 3 is used, the peak 7 appears near the end of the gradient, and when similar situations occur, the time can be appropriately prolonged to ensure that the obtained chromatogram is complete.
3. Methodology investigation
3.1 repeatability
Taking 6 parts (KL-QH-14) of the same Notopterygii rhizoma sample, preparing a sample solution according to the method of example 7, determining to obtain a characteristic spectrum, calculating relative peak area and relative retention time by using the No. 2 peak (S1) and the No. 4 peak (S2) as reference peaks, and calculating RSD value, the result is shown in Table 18.
TABLE 18 relative retention time of characteristic peaks
According to the repeatability inspection result, the relative retention time RSD value of each characteristic peak is in the range of 0.0% -0.1%, and the characteristic spectrum is good in repeatability.
3.2 intermediate precision
Different analysts adopt Waters UPLC H-Class and PDA detector at different time, take notopterygium root sample (KL-QH-14), prepare sample solution according to the method of example 7, obtain characteristic spectrum, take No. 2 peak (S1) and No. 3 peak (S2) as reference peak, calculate relative retention time of characteristic peak, the result is shown in Table 19.
Table 19 examines the relative retention times of characteristic peaks
From the results, the RSD value of the relative retention time of each characteristic peak of the sample between different instruments is in the range of 0.0-0.1%, which shows that the intermediate precision of the characteristic spectrum method is good
3.3 stability
A Notopterygii rhizoma sample (KL-QH-14) is taken, a sample solution is prepared according to the method of example 7, the characteristic spectrum is obtained by measuring according to the method of example 7 at 0, 2, 4, 6, 8, 10, 12 and 24h respectively, and the RSD is calculated, and the result is shown in Table 20.
TABLE 20 relative retention time and float Range of characteristic peaks
By investigating the stability of the solution for 24 hours, the relative retention time of each characteristic peak is within +/-10% of a specified value, so that the stability of the characteristic spectrum analysis method of the notopterygium root standard decoction adopted by the invention is better.
Example 8
The embodiment provides a quality detection method of notopterygium root formula particles, which comprises the following steps,
(1) Preparation of control solutions: taking an appropriate amount of decursin reference substance, precisely weighing, and adding methanol to obtain reference substance solution E containing 200 μ g per 1 ml;
(2) Preparation of a test solution: taking 0.2g of each of three batches of notopterygium root formula particles (batch numbers: KL-QH-14, KL-QH-15 and KL-QH-16), precisely weighing, placing in a conical flask with a stopper, precisely adding 20ml of 70% methanol, weighing, ultrasonically treating (power 250W and frequency 40 kHZ) for 30min, cooling, weighing again, complementing the lost weight with 70% methanol, shaking up, filtering, and taking the obtained filtrate as a test solution.
(3) Performing high performance liquid chromatography with octadecylsilane chemically bonded silica as filler (Agilent ZORBAX Eclipse XDB-C18.6 × 250mm,5 μm), methanol-water (40) as mobile phase, detection wavelength of 334nm, column temperature of 30 deg.C, and flow rate of 1.0ml/min; the sample injection amount is 10 mu L;
through detection, the contents of decursin in the three batches of notopterygium root formula granules (KL-QH-14, KL-QH-15 and KL-QH-16) are respectively 41.13mg, 6.69mg and 24.61mg, and the quality of the notopterygium root formula granules prepared by the method meets the requirements within the specified value of 4-35 mg.
Wherein, the calculation formula of the content of decursin is as follows:
wherein, W Test article The peak area, C, of decursin in the characteristic map of the sample Reference substance Represents the concentration of the control solution, V Reference substance Represents the volume of the control solution, W Reference substance Peak area, C, representing decursin control Test article Represents the concentration of the test solution, V Test article Represents the volume of the control solution.
Experimental example 2
1. Examination of preparation of test solution
1.1 selection of extraction solvent
The extraction solvent is water, methanol, 70% methanol, 50% methanol, 30% methanol, ethanol, 70% ethanol, 50% ethanol, and 30% ethanol. The preparation method of the test solution comprises the following steps: grinding a plurality of same test samples (KL-QH-14), precisely weighing about 0.2g, placing in a conical flask with a plug, adding 20ml of extraction solvent, sealing the plug, weighing, carrying out ultrasonic treatment for 30min (power 250W and frequency 40 kHz), taking out, cooling, weighing again, adding the extraction solvent to complement weight loss, shaking up, and filtering to obtain a test sample solution. Precisely sucking 2 mu L of test solution, injecting into an ultra-high performance liquid chromatograph, measuring according to the chromatographic conditions of the embodiment 8, obtaining results shown in a table 21, measuring each sample twice, and taking an average value;
TABLE 21 content of decursin in test samples obtained by different extraction solvents
The extraction rate of 70% methanol as extraction solvent is high, the content of decursin is highest, the result is consistent with the characteristic map construction method, and 70% methanol is preferably used as extraction solvent.
1.2 selection of extraction mode
Taking an extraction mode as a variable, respectively extracting and preparing a sample solution by adopting two methods of ultrasonic extraction and heating reflux extraction, wherein the specific method comprises the following steps: taking a proper amount of the same test sample (KL-QH-14), grinding, taking 0.2g, precisely weighing, placing in a conical flask with a plug, precisely adding 20ml of 70% methanol, sealing the plug, weighing, respectively heating and refluxing (85 ℃, heating in a water bath) and treating with ultrasound (power 250W and frequency 40 kHZ) for 30min, taking out, cooling, complementing the lost weight, shaking up, filtering, taking out the subsequent filtrate, obtaining the content of decursin according to the method of the embodiment 8, measuring each sample twice, and taking the average value, wherein the results are shown in a table 22.
TABLE 22 different extraction methods to obtain the content of decursin in the test solution
The content difference of decursin measured by ultrasonic extraction and reflux extraction is not big, the ultrasonic extraction operation is convenient, the energy is saved, and the ultrasonic extraction is preferred.
1.3 selection of the amount of extraction solvent added
Taking the addition amount of the extraction solvent as a variable, the addition amounts of the extraction solvent are respectively 10ml, 20ml and 50ml, the test sample solution is prepared according to the method in section 1.1 of the embodiment, the content of decursin is obtained according to the determination of the chromatographic conditions in embodiment 8, each sample is determined twice, and the average value is obtained, and the result is shown in table 23.
TABLE 23 the content of decursin in the test solutions obtained by adding different extraction solvents
When the solvent amount is 10ml, the extraction of the decursin is incomplete, and when the solvent amount is 20ml and 50ml, the extraction content of the decursin is not greatly different, which indicates that the extraction is complete. In order to save solvent, the amount of extraction solvent was selected to be 20ml.
1.4 examination of extraction time
Taking the ultrasonic extraction time as a variable, wherein the ultrasonic extraction time is respectively 20min, 30min and 40min, preparing a test solution according to the section 1.1 of the experimental example, measuring according to the chromatographic conditions of the example 8 to obtain the content of decursin, measuring each sample twice, and taking an average value, wherein the result is shown in a table 24.
TABLE 24 determination of the content of decursin in the test solutions obtained at different extraction times
The extraction time is 30min, the content of decursin is higher than that of 20min and 40min, which indicates that the extraction time is 30min, the decursin can be completely extracted, therefore, the extraction time is preferably 30min.
In summary, the preparation method of the test solution comprises the following steps: taking a proper amount of a test sample, grinding, taking about 0.2g, precisely weighing, placing in a conical flask with a plug, precisely adding 20ml of 70% methanol, sealing the plug, weighing, carrying out ultrasonic treatment (power 250W, frequency 40 kHz) for 30 minutes, taking out, cooling, supplementing the weight loss with 70% methanol, shaking up, filtering, and taking a subsequent filtrate to obtain the product.
2. Selection of chromatographic conditions
2.1 selection of detection wavelength
The decursin reference solution prepared according to the method of example 8 was detected by a diode array detector, and the maximum absorption wavelength of decursin was 334nm, as shown in fig. 11.
2.2 Peak purity experiment
Notopterygium incisum sample solution is taken, the preparation method is the same as that of example 8, and the diode array detector is utilized to carry out peak purity inspection on decursin, as shown in figure 12 and table 25.
TABLE 25 purity test of decursin
| | Purity | 1 | Purity | 1 threshold |
| 8.040 | 0.648 | 7.740 |
In the chromatogram of the test sample, the peak purity angle of decursin is less than the purity threshold value of 0.648 and less than 7.740, which indicates that the purity of decursin obtained by the method meets the analysis requirement.
2.3 selection of column temperature
The contents of decursin in the test solutions were determined twice for each sample by measuring the column temperature as a variable at 28 ℃,30 ℃ and 32 ℃ according to the method of example 8, and the results are shown in table 26 and fig. 13.
TABLE 26 content of decursin in test solutions at different column temperatures
From the above results, it can be seen that there is no significant difference in the measured results at different column temperatures, and the RSD is 1.7%, so that the durability of the method to the column temperature is satisfactory.
2.4 selection of flow Rate
Taking the flow rates as variables, the flow rates are respectively 0.8ml/min, 1.0ml/min and 1.2ml/min, the rest is the same as example 8, the same notopterygium root test sample solution is taken and measured at different flow rates, the content of decursin in the test sample solution is obtained, each sample is measured twice, and the average value is taken, which is shown in figure 14 and table 27.
TABLE 27 content of decursin in test solutions at different flow rates
From the above results, the results measured at different flow rates were not significantly different, and RSD was 1.6%, so the durability of the method to column temperature was satisfactory.
2.5 investigation of the column
Taking the chromatographic columns as variables, and taking the same notopterygium root test sample solution as in example 8 for the rest, and measuring under different chromatographic columns to obtain the content of decursin in the test sample solution, table 28.
A chromatographic column 1: agilent ZORBAX Eclipse XDB-C18 (4.6X 250mm,5 μm)
And (3) chromatographic column 2: agilent ZORBAX extended C18 (4.6X 250mm,5 μm)
And (3) chromatographic column: waters Xbridge C18 (4.6X 250mm,5 μm)
TABLE 28 content of decursin in test solutions measured by different chromatographic columns
From the results, the RSD of the decursin content result under the chromatographic column conditions of different manufacturers and types is 2.0 percent, and the RSD meets the requirement of system applicability. The method is shown to have better durability to different chromatographic columns.
3. Methodology investigation
3.1 repeatability test
The content of decursin in the test sample solution was determined by the method of example 8 using 6 parts (KL-QH-14) of the same Notopterygii rhizoma test sample, as shown in Table 29.
TABLE 29 results of repeated experiments
According to the determination results, the average content of the decursin in the sample is 26.0mg/g, the RSD is 1.4%, the repeatability is good, and the analysis requirements are met.
3.2 intermediate precision
Different analysts performed intermediate precision tests at different times using another Agilent 1260 definition high performance liquid chromatograph (TUV detector), column: agilent ZORBAX Eclipse XDB-C18 (4.6X 250mm,5 μm), test solution of Notopterygii rhizoma (KL-QH-14) was prepared according to example 8, and the content of decursin was determined, as shown in Table 30.
TABLE 30 intermediate precision results
From the above results, the content of decursin in the test sample is 1.1%, and the content of decursin measured by different instruments is 1.7%, which meets the analysis requirements.
3.3 stability
A test solution of Notopterygii rhizoma (KL-QH-14) was prepared according to the method of example 8, and the sample was measured at 0, 2, 4, 6, 8, 10, 12, and 24h, respectively, with a sample size of 10 μ L, to obtain the peak area of the decursin peak in the test solution, and the RSD was calculated, and the results are shown in Table 31.
TABLE 31 stability test results
Within 24h, the RSD of the area of decursin is 0.9%, which indicates that the test solution has good stability and meets the analysis requirement.
3.4 accuracy
Accurately weighing appropriate amount of decursin reference substance, placing in 100ml volumetric flask, adding 75% methanol to dissolve, fixing volume to scale, shaking, and making into reference substance solution containing 0.2640mg of decursin per 1ml for use. 9 parts of test solution with known content of decursin under item 3.2, each part is about 0.1g, and the mass is precisely weighed respectively as shown in table 32. Precisely adding 5ml of decursin reference solution into 3 parts of test solution, and then adding 15ml of 75% methanol to obtain samples a-c; precisely adding 10ml of decursin reference solution into another 3 parts of test solution, and then adding 10ml of 75% methanol to obtain samples d-f; taking 3 parts of test solution, respectively and precisely adding 15ml of decursin reference solution, then adding 5ml of 75% methanol to obtain samples g-i, respectively, measuring the content of decursin in the samples a-i according to the example 8, calculating the recovery rate, and referring to the following formula for the calculation formula of the recovery rate, the result is shown in table 32.
TABLE 32 experiment on recovery of decursin
According to the recovery rate test result, the content of the decursin is measured, the recovery rate ranges from 99.41% to 105.10%, the average recovery rate is 102.7%, the RSD value is 1.8%, and the analysis requirement is met.
3.5 determination of the Linear equation
Preparing a decursin reference solution: accurately weighing 4.125mg of decursin reference substance, placing in a 10ml volumetric flask, adding 70% methanol for dissolving, fixing the volume to scale, shaking up, and marking as reference substance solution A;
precisely sucking 5ml of the reference substance solution A, placing the reference substance solution A into a 10ml volumetric flask, adding 70% methanol to a constant volume to a scale, shaking up, filtering, and marking as a reference substance solution B;
precisely sucking 5ml of the reference solution B, placing in a 10ml volumetric flask, adding 70% methanol to constant volume to scale, shaking up, filtering, and marking as decursin C;
precisely sucking 1ml of decursin C, placing in a 10ml volumetric flask, adding 70% methanol to a constant volume to a scale, shaking up, filtering, and marking as decursin D;
precisely sucking 5ml of decursin D, placing into a 10ml volumetric flask, adding 70% methanol to a constant volume to a scale, shaking up, filtering, and marking as decursin E.
Precisely absorbing 10 mu l of each of the decursin reference substance solutions A-E, injecting into a high performance liquid chromatograph, measuring the peak area of the decursin chromatographic peak according to the conditions and methods under the content measurement item, taking the peak area of the decursin chromatographic peak as a vertical coordinate and the concentration of the decursin as a horizontal coordinate, performing linear regression, and obtaining a regression equation (including an origin) of y =3E +07x +29259 and R2=1, wherein the linear range is 0.005-0.40mg/ml. Quantification can be done using the external standard one-point method. See table 33 and fig. 15.
TABLE 33 Philippine Swingoside reference solution concentration and peak area
| Number of | E | D | C | B | A |
| Decursin concentration (mg/ml) | 0.005022 | 0.010044 | 0.100444 | 0.200888 | 0.4018 |
| Area of nod of decursin | 146017 | 291493.5 | 2848772 | 5608439 | 11135692 |
Example 9
The embodiment provides a method for constructing a characteristic spectrum of notopterygium root decoction pieces, which comprises the following steps,
(1) Preparation of control solutions: notopterygium incisum alcohol reference substance solution A, isoimperatorin reference substance solution B, ferulic acid reference substance solution C and chlorogenic acid reference substance solution D are prepared according to the method provided by the embodiment 7.
(2) Preparation of reference solution of reference drug: notopterygium incisum reference drug solution is prepared according to the method provided in example 7.
(3) Preparation of a test solution: pulverizing Notopterygii rhizoma decoction pieces (lot number: YP-QH-14) into powder, collecting Notopterygii rhizoma decoction pieces 0.4g, precisely weighing, placing in a conical flask with a plug, precisely adding 70% methanol 50ml, weighing, ultrasonic treating (power 250W, frequency 40 kHZ) for 30min, cooling, weighing again, supplementing the weight loss with 70% methanol, shaking, filtering, and collecting the filtrate as sample solution.
(4) The method provided in example 7 is followed to obtain the characteristic maps of the notopterygium root decoction pieces test solution, the notopterygium root reference medicinal material solution and the reference solution, respectively, and the characteristic maps are shown in FIG. 16.
It should be understood that the above examples are only for clarity of illustration and are not intended to limit the embodiments. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. And obvious variations or modifications of the invention may be made without departing from the spirit or scope of the invention.
Claims (14)
1. A method for constructing a characteristic spectrum of notopterygium root decoction pieces or formula granules is characterized by comprising the following steps of,
(1) Preparing a test solution;
(2) Detecting a sample solution by adopting ultra-high performance liquid chromatography, taking octadecylsilane chemically bonded silica as a filling agent, and adopting the following specifications: 2.1X 100mm,1.7 μm, acetonitrile as mobile phase A, phosphoric acid containing solution as mobile phase B, gradient elution, the gradient elution program includes: 0-6min, the volume percentage of the mobile phase A is 10-15%, and the volume percentage of the mobile phase B is 90-85%;6-9min, the volume percentage of the mobile phase A is 15-16%, and the volume percentage of the mobile phase B is 85-84%;9-14min, the volume percentage of the mobile phase A is 16-47%, and the volume percentage of the mobile phase B is 84-53%;14-18min, wherein the volume percent of the mobile phase A is 47-53%, and the volume percent of the mobile phase B is 53-47%;18-21min, wherein the volume percent of the mobile phase A is 53-90%, and the volume percent of the mobile phase B is 47-10%;
wherein the extraction solvent for preparing the test solution is 70% methanol, 50% methanol or 30% methanol.
2. The method according to claim 1, further comprising a step of preparing a reference solution from at least one of notopterygium alcohol, isoimperatorin, ferulic acid, chlorogenic acid, decursin and phenethyl ferulate, and a step of detecting the reference solution by ultra-high performance liquid chromatography in the method according to claim 1 to obtain a reference characteristic map.
3. The construction method according to claim 2, wherein the preparation method of the reference solution comprises the following steps: taking notopterygium alcohol, adding solvent, and making into notopterygium alcohol control solution A containing 10-20 μ g of notopterygium alcohol per 1 ml;
adding solvent into isoimperatorin to obtain isoimperatorin reference solution B containing 5-15 μ g of isoimperatorin per 1 ml;
adding solvent into ferulic acid, and making into ferulic acid control solution C containing 30-50 μ g per 1 ml;
adding solvent into chlorogenic acid, and making into chlorogenic acid reference solution D containing 50-70 μ g of chlorogenic acid per 1 ml;
collecting decursin, adding solvent, and making into decursin control solution E containing 150-250 μ g per 1 ml;
taking the ferulic acid phenethyl alcohol ester, adding a solvent, and preparing a reference substance solution F containing 30-80 mu g of ferulic acid phenethyl alcohol ester per 1 ml;
the solvent is selected from methanol aqueous solution, ethanol aqueous solution or water; the volume fraction of methanol in the methanol aqueous solution is 60-70%.
4. The constructing method according to claim 1 or 2, wherein the preparation of the test solution comprises the steps of,
extracting Notopterygii rhizoma decoction pieces powder or Notopterygii rhizoma granule to obtain sample solution.
5. The method according to claim 4, further comprising extracting a reference solution of Notopterygium incisum from a Notopterygium incisum reference drug.
6. The construction method according to claim 1 or 2, wherein the chromatographic conditions of the ultra high performance liquid chromatography further comprise: the detection wavelength is 280-300nm; the flow rate is 0.2-0.6ml/min; the column temperature is 38-42 ℃; the sample amount is 1-5 muL;
taking phosphoric acid water solution with volume fraction of 0.05-0.15% as mobile phase B.
7. The constructing method according to claim 1 or 2, wherein the Notopterygium incisum formula particle feature map is selected from any one of the following (1) to (2);
(1) The characteristic map at least comprises 5 characteristic peaks, wherein the peak 2 is chlorogenic acid, the peak 3 is ferulic acid, the peak 5 is decursin, and the relative retention time of each characteristic peak is within +/-10% of a specified value;
taking peak No. 2 as a reference peak, and the relative retention time of peak No. 1 is 0.79;
taking the peak No. 3 as a reference peak, and the relative retention time of the peak No. 4 is 1.08;
(2) The characteristic map at least comprises 8 characteristic peaks, wherein the peak 2 is chlorogenic acid, the peak 3 is ferulic acid, the peak 5 is decursin, the peak 6 is notopterygium alcohol, the peak 7 is phenethyl ferulate, and the peak 8 is isoimperatorin; the relative retention time of each characteristic peak is within +/-10% of a specified value;
taking peak No. 2 as a reference peak, and the relative retention time of peak No. 1 is 0.79;
the relative retention time of peak No. 4 was 1.08 with peak No. 3 as the reference peak.
8. The method of claim 1 or 2, wherein the Notopterygium incisum formula particles are prepared by a process comprising the steps of,
(1) Processing Notopterygii rhizoma to obtain Notopterygii rhizoma decoction pieces;
(2) Decocting Notopterygii rhizoma decoction pieces to obtain Notopterygii rhizoma extractive solution, and concentrating to obtain fluid extract with relative density of 1.05-1.15 g/ml;
(3) Directly spray-drying the clear paste to obtain dry powder;
(4) And mixing the dry powder with auxiliary materials, and granulating to obtain the notopterygium root formula granules.
9. The method of construction according to claim 8, wherein the inlet air temperature for spray drying is 165-185 ℃.
10. The constructing method of claim 8, wherein the decocting comprises adding water 8-12 times the amount of decoction pieces to the first decoction, boiling for 30-60min, adding water 8-10 times the amount of decoction pieces to the second decoction, boiling for 30-60min, and mixing the filtrates to obtain the Notopterygii rhizoma extract.
11. The constructing method according to claim 10, wherein the extract yield of the Notopterygium incisum extract is 16-25%;
the processing steps comprise taking notopterygium root medicinal materials, spraying water, moistening for at least 12h, and drying at low temperature to obtain notopterygium root decoction pieces.
12. The constructing method according to claim 1 or 2, wherein the quality testing method of Notopterygium incisum formula particles comprises,
(1) Preparing a reference substance solution E by adopting a decursin reference substance;
(2) Preparing a test solution;
(3) Performing high performance liquid chromatography, eluting with methanol and water as mobile phase at equal rate to obtain test solution containing decursin 2-32mg.
13. The method of claim 12, wherein the chromatographic conditions further comprise a detection wavelength of 330-340nm; the flow rate is 0.8-1.2ml/min; the column temperature is 28-32 ℃; the sample amount is 8-12 μ L.
14. The method for constructing a Chinese medicinal composition according to claim 12, wherein the control solution E contains about 150-250 μ g of decursin per 1ml of the control solution E.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111070341.2A CN113759045B (en) | 2021-09-13 | 2021-09-13 | Characteristic spectrum of notopterygium root decoction pieces or formula granules, construction method of characteristic spectrum, notopterygium root formula granules, preparation method of notopterygium root formula granules and quality control method of notopterygium root formula granules |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111070341.2A CN113759045B (en) | 2021-09-13 | 2021-09-13 | Characteristic spectrum of notopterygium root decoction pieces or formula granules, construction method of characteristic spectrum, notopterygium root formula granules, preparation method of notopterygium root formula granules and quality control method of notopterygium root formula granules |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113759045A CN113759045A (en) | 2021-12-07 |
| CN113759045B true CN113759045B (en) | 2023-03-31 |
Family
ID=78795308
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111070341.2A Active CN113759045B (en) | 2021-09-13 | 2021-09-13 | Characteristic spectrum of notopterygium root decoction pieces or formula granules, construction method of characteristic spectrum, notopterygium root formula granules, preparation method of notopterygium root formula granules and quality control method of notopterygium root formula granules |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113759045B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114636779B (en) * | 2022-03-29 | 2024-05-24 | 陕西盘龙药业集团股份有限公司 | Construction method of three-conversion soup reference sample freeze-dried powder fingerprint and fingerprint thereof |
| CN116242935B (en) * | 2023-02-03 | 2024-04-26 | 广东一方制药有限公司 | Construction method of characteristic patterns of notopterygium root or notopterygium latifolium and identification method thereof |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101474290B (en) * | 2007-12-18 | 2012-07-25 | 北京康仁堂药业有限公司 | Detection method of figwort root dispensing granule |
| CN108771692A (en) * | 2018-07-06 | 2018-11-09 | 徐漫 | Rhizoma Et Radix Notopterygii is fresh to carry spray-dried powders medicine materical crude slice |
| CN108524563A (en) * | 2018-07-06 | 2018-09-14 | 徐漫 | Rhizoma Et Radix Notopterygii is fresh to carry vacuum drying powder decoction pieces |
-
2021
- 2021-09-13 CN CN202111070341.2A patent/CN113759045B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN113759045A (en) | 2021-12-07 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109828059B (en) | Detection method of cassia twig, peony and rhizoma anemarrhenae decoction | |
| CN101850070B (en) | Detection method for Chinese medicament Tangcao tablets | |
| CN113759045B (en) | Characteristic spectrum of notopterygium root decoction pieces or formula granules, construction method of characteristic spectrum, notopterygium root formula granules, preparation method of notopterygium root formula granules and quality control method of notopterygium root formula granules | |
| CN112587642B (en) | Preparation method and detection method of vitality-maintaining pharmaceutical composition | |
| CN109342631B (en) | Method for constructing HPLC fingerprint of Chinese medicinal composition and method for detecting quality of Chinese medicinal composition | |
| CN108362788A (en) | A kind of steamed sealwort Quality Detection analysis method | |
| CN108459128B (en) | Quality control method of angelica sinensis Sini decoction composition | |
| CN113533614B (en) | Method for establishing material standard of Xiaoqi decoction | |
| CN113777183A (en) | Method for constructing characteristic spectrum of glossy privet fruit medicinal material and processed product thereof and method for detecting content of multi-index components | |
| CN112798701A (en) | Preparation method and detection method of angelica sinensis blood-enriching pharmaceutical composition | |
| CN115575551B (en) | Bletilla striata detection method | |
| CN110464825A (en) | A kind of dihuang drink pharmaceutical composition, preparation method, detection method | |
| CN107764924B (en) | Detection method of effective components in asthma granules | |
| CN113759038B (en) | Trachelospermi caulis characteristic map, construction method and quality control method thereof, trachelospermi caulis formula granules and preparation method thereof | |
| CN113759011B (en) | Method for establishing characteristic spectrum of starwort root and preparation thereof | |
| CN115508463A (en) | Detection method and quality control method of Tanggute radix Et rhizoma Rhei product | |
| CN114994220A (en) | Construction method of fingerprint of Qiqing toxin-vanquishing granules, determination method of component content of Qiqing toxin-vanquishing granules and application of Qiqing toxin-vanquishing granules | |
| CN112098569B (en) | Preparation method and quality determination method of radix clematidis extract | |
| CN112763639A (en) | Preparation process and quality control method of radix Acanthopanacis Senticosi reference extract | |
| CN113759043B (en) | Characteristic spectrum of motherwort fruit and construction method thereof, method for measuring content of stachydrine hydrochloride, motherwort fruit formula granules and preparation method thereof | |
| CN113759040B (en) | Cat's claw grass and preparation characteristic map and construction method thereof, and method for measuring content of cat's claw grass and preparation thereof | |
| CN115266961B (en) | Construction method of characteristic spectrum of perilla stem medicinal preparation | |
| CN113759036B (en) | Method for measuring content of protodioscin in rhizoma Dioscoreae Septemlobae | |
| CN109709250B (en) | Method for detecting fingerprint of ginseng and pilose antler wine | |
| CN113759026B (en) | Common clubmoss herb and preparation characteristic map and construction method thereof |
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 |