CN110455973B - Method for establishing fingerprint spectrum of Tou Tang and method for controlling quality of Tou Tang - Google Patents
Method for establishing fingerprint spectrum of Tou Tang and method for controlling quality of Tou Tang Download PDFInfo
- Publication number
- CN110455973B CN110455973B CN201910772761.1A CN201910772761A CN110455973B CN 110455973 B CN110455973 B CN 110455973B CN 201910772761 A CN201910772761 A CN 201910772761A CN 110455973 B CN110455973 B CN 110455973B
- Authority
- CN
- China
- Prior art keywords
- soup
- fingerprint
- sample
- parts
- acetonitrile
- 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
- 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
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/34—Control of physical parameters of the fluid carrier of fluid composition, e.g. gradient
-
- 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
-
- 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
- G01N30/86—Signal analysis
- G01N30/8675—Evaluation, i.e. decoding of the signal into analytical information
- G01N30/8686—Fingerprinting, e.g. without prior knowledge of the sample components
Landscapes
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Engineering & Computer Science (AREA)
- Library & Information Science (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
Abstract
A method for establishing finger print of Tou Zong Tang and a method for controlling quality of Tou Zong Tang belong to the technical field of liquid chromatography. A method for establishing the fingerprint of the Zodiac soup comprises the steps of detecting the fingerprint of a Zodiac soup sample by using a high performance liquid chromatograph, selecting acetonitrile and water as mobile phases during detection, and performing gradient elution, wherein the flow rate of the mobile phases is 0.6-1.1 mL/min, and the volume ratio of the acetonitrile in the mobile phases is 2% -40%. According to the method, acetonitrile and water are used as mobile phases to carry out gradient elution on the soup sample, so that various substance components in the soup sample can be accurately separated, and the complete fingerprint with an obvious characteristic peak pattern is obtained. The method for controlling the quality of the tape finishing soup determines whether the tape finishing soup is changed or not by comparing the fingerprint spectrums of the extracting solution, the concentrated solution, the dry paste and the finished product particle sample of different batches of the tape finishing soup in the preparation process, thereby achieving the purpose of controlling the quality of the tape finishing soup in the preparation process.
Description
Technical Field
The application relates to the technical field of liquid chromatography, in particular to a method for establishing a fingerprint of a Wangtiantang and a quality control method of the Wangtiantang.
Background
The Belt eliminating decoction is a dampness eliminating agent and has the effects of tonifying spleen, soothing liver, eliminating dampness and stopping leukorrhagia. It is mainly used for treating spleen deficiency, stagnation of liver-qi, damp turbidity and leukorrhagia. A white leucorrhea, thin and clear nasal discharge and a pale complexion
White, lassitude, loose stool, pale tongue with white coating, and slow or weak pulse.
The components of the traditional Chinese medicine compound are very complex, the components are mutually influenced, the water extract of the traditional Chinese medicine compound has high polarity, and a plurality of substances can be effectively extracted, so that the sufficient separation of the substances is difficult to ensure, and the quality control difficulty is increased.
Disclosure of Invention
The application provides a method for establishing a fingerprint of a Wangtian soup and a method for controlling the quality of the Wangtian soup, which can effectively separate various effective components in the Wangtian soup by using a liquid chromatograph to obtain a complete fingerprint so as to achieve the purpose of controlling the quality.
The embodiment of the application is realized as follows:
in a first aspect, the present examples provide a finishing soup fingerprint spectrum establishment method, which includes detecting a fingerprint spectrum of a finishing soup sample by using a high performance liquid chromatograph;
during detection, acetonitrile and water are selected as mobile phases, gradient elution is adopted, and the flow rate of the mobile phases is 0.6-1.1 mL/min;
the volume proportion of acetonitrile in the mobile phase is 2-40%;
the TAIBAI decoction is prepared from Atractylodis rhizoma, rhizoma Dioscoreae, Ginseng radix, radix Paeoniae alba, semen plantaginis, rhizoma Atractylodis, Glycyrrhrizae radix, pericarpium Citri Tangerinae, herba Schizonepetae and bupleuri radix.
In the technical scheme, the fingerprint of the sample of the Belt finalization soup is detected by a high performance liquid chromatograph, the Belt finalization soup is prepared from raw materials including rhizoma atractylodis macrocephalae, Chinese yam, ginseng, radix paeoniae alba, semen plantaginis, rhizoma atractylodis, liquorice, dried orange peel, black schizonepeta spike and radix bupleuri, and the belt finalization soup contains various active substance components. According to the method, acetonitrile and water are used as mobile phases to carry out gradient elution on the soup sample, so that various substance components in the soup sample can be accurately separated, and the complete fingerprint with an obvious characteristic peak pattern is obtained.
In a first possible example of the first aspect of the present application in combination with the first aspect, the chromatography column used for the above detection is a C18 water-resistant chromatography column.
In the above example, the chromatographic column facilitates the high performance liquid chromatograph to accurately and completely separate out various substance components in the tape-out soup, and the various substance components are displayed in a map.
In a second possible example of the first aspect of the present application in combination with the first aspect, the column temperature of the column during the detection is 30 to 40 ℃.
In the above example, the column temperature detection of the chromatographic column is beneficial for the high performance liquid chromatograph to accurately and completely separate out various substance components in the strip soup, and the various substance components are displayed in a map.
With reference to the first aspect, in a third possible example of the first aspect of the present application, the detection wavelength of the ultraviolet detector is set to 240 to 250nm when the liquid-phase detection is performed.
In the above example, the set detection wavelength of the ultraviolet detector obtained through analysis and experiments is adapted to various active substance components in the tape finishing soup, that is, the absorption wavelength of the active substance components in the tape finishing soup is 240-250 nm, which can ensure that the active substance components in the tape finishing soup can present specific peaks in the spectrum.
In a fourth possible example of the first aspect of the present application in combination with the first aspect, when performing the gradient elution described above, the elution is performed in the following manner:
in 0-10 min, the volume proportion of acetonitrile in the mobile phase is increased from 2% to 6%;
in 10-40 min, the volume proportion of acetonitrile in the mobile phase is increased from 6% to 15%;
in 40-60 min, the volume ratio of acetonitrile in the mobile phase is 15%;
increasing the volume proportion of acetonitrile in the mobile phase from 15% to 20% in 60-80 min;
and in 80-100 min, the volume proportion of the acetonitrile in the mobile phase is increased from 20% to 40%.
In the above example, the mobile phase is eluted in the above manner, which facilitates the separation of various active substance components in the finished soup, so that they are completely and clearly displayed in the subsequently presented map.
In a fifth possible example of the first aspect of the present application, in combination with the first aspect, the gradient elution is performed at a flow rate of the mobile phase of 0.6 to 0.8mL/min for 0 to 10min and at a flow rate of 0.9 to 1.1mL/min for 10 to 100 min.
In the above example, the elution of the mobile phase at the above flow rate is advantageous for separating various active substance components in the complete strip soup, and the mobile phase is not wasted.
In a sixth possible example of the first aspect of the present application, in combination with the first aspect, when the gradient elution is performed, the flow rate of the mobile phase is 0.7mL/min in 0 to 10min, and the flow rate of the mobile phase is 1.0mL/min in 10 to 100 min.
In a second aspect, the present application provides a method for quality control of a take-up soup, which includes taking an extract sample, a concentrated solution sample, a dry paste sample and a finished product particle sample of different batches of the take-up soup during a preparation process respectively, and detecting fingerprint spectrums by using the above-mentioned method for establishing a take-up soup fingerprint spectrum, wherein all fingerprint spectrums at least include 13 common chromatographic peaks, and the similarity of all fingerprint spectrums is greater than or equal to 97%.
In the technical scheme, the fingerprint spectrums of the extracting solution, the concentrated solution, the dry paste and the finished product particle sample in the preparation process of different batches of the soup for finishing the carrying are compared, and the number and the height of the common chromatographic peaks are compared to determine whether the effective substance components in the extracting solution sample, the concentrated solution sample, the dry paste sample and the finished product particle sample in the preparation process of the soup for finishing the carrying are the same or not, and whether the effective substance components in the soup for finishing the carrying and the different batches are changed or not, so that the quality of the soup for finishing the carrying in the preparation process is controlled.
In a first possible example of the first aspect of the present application, in combination with the second aspect, the above-mentioned tanga soup is prepared in the following manner:
according to parts by weight, 30-40 parts of soil-fried bighead atractylodes rhizome, 30-40 parts of fried Chinese yam, 5-10 parts of ginseng, 15-20 parts of wine-fried white paeony root and 10-15 parts of wine-fried plantain seed are added; 10-15 parts of rhizoma atractylodis; 3-5 parts of liquorice; adding water into 1-2 parts of pericarpium citri reticulatae, 1-2 parts of black schizonepeta spike and 2-3 parts of radix bupleuri, heating and extracting for 2-3 times, each time for 0.5-2 hours, combining extracting solutions, filtering, concentrating to obtain a concentrated solution, drying to obtain a dry paste, and adding auxiliary materials to prepare a finished granular traditional Chinese medicine preparation.
In the above examples, the Tokuai decoction is prepared by the above method, wherein the main active ingredients in the medicinal materials can be effectively preserved without change, and the prepared Tokuai decoction has good curative effect.
In a second possible example of the first aspect of the present application in combination with the second aspect, the above-mentioned extract liquid sample, concentrated liquid sample, dried paste sample and finished granule sample are obtained by:
filtering the extracting solution to obtain an extracting solution sample;
mixing 4g of concentrated solution with water, fixing the volume to 50mL, and filtering to obtain a concentrated solution sample;
mixing 1g of dry paste with water, diluting to 50mL, and filtering to obtain a dry paste sample;
and (3) mixing 1g of finished product particles with water, metering the volume to 50mL, and filtering to obtain a finished product particle sample.
In the above example, the extract sample, the concentrated solution sample, the dry paste sample and the finished product particle sample are prepared by the above method, so that the concentration difference of the extract sample, the concentrated solution sample, the dry paste sample and the finished product particle sample is ensured to be small, and the subsequent comparison of the characteristic maps is facilitated.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings that are required to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present application and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained from the drawings without inventive effort.
FIG. 1 shows HPLC finger prints obtained in example 1 and comparative examples 1 and 2 of the present application;
FIG. 2 is a HPLC fingerprint obtained in example 2 of the present application;
FIG. 3 shows the HPLC fingerprint obtained in example 3 of the present application.
Detailed Description
Embodiments of the present application will be described in detail below with reference to examples, but those skilled in the art will appreciate that the following examples are only illustrative of the present application and should not be construed as limiting the scope of the present application. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
High Performance Liquid Chromatography (High Performance Liquid Chromatography, HPLC) is an important branch of Chromatography, in which Liquid is used as a mobile phase, a High-pressure infusion system is adopted, mobile phases such as single solvents with different polarities or mixed solvents, buffers and the like with different proportions are pumped into a chromatographic column filled with a stationary phase, and after components in the column are separated, the mobile phases enter a detector for detection, so that analysis of a sample is realized.
However, the method is different when HPLC is used to detect different mixed liquids. When the detection method is selected unreasonably, the liquid separation is not thorough, and the peak appearance of the detected spectrum is not obvious.
The following description specifically describes a finishing soup fingerprint spectrum establishing method and a finishing soup quality control method according to embodiments of the present application:
the application provides a method for establishing a fingerprint of a Wantangtang, which is used for detecting a Wantangtang sample by an HPLC method.
The Wangbai decoction is prepared from raw materials including white atractylodes rhizome, Chinese yam, ginseng, white paeony root, plantain seed, rhizoma atractylodis, liquorice, dried orange peel, black schizonepeta spike and Chinese thorowax root.
According to the effective substance ingredients in the traditional Chinese medicine ingredients, the interference situation among different substances needs to be analyzed and eliminated. The selection of a proper chromatographic column and a proper mobile phase and the control of the flow rate of the mobile phase are favorable for separating various effective substance components in the finished soup, so that the various effective substance components can present specific peaks in a map to obtain accurate map data.
According to the method, 10-20 mu L of sample is sucked for HPLC detection, acetonitrile and water are selected as mobile phases, the volume ratio of the acetonitrile in the mobile phases is 2% -40%, gradient elution is adopted, and the flow rate of the mobile phases is 0.6-1.1 mL/min.
Optionally, the measurement wavelength of the ultraviolet detector is set to 240-250 nm. The wavelength can be used for various active substance components in the tape finishing soup, namely the absorption wavelength of the active substance components in the tape finishing soup is 240-250 nm, and the active substance components in the tape finishing soup can be ensured to present specific peaks in a map.
The ultraviolet detector can be self-contained or externally connected with the liquid chromatograph.
Optionally, the chromatographic column selected for the application is a C18 water-resistant chromatographic column, and the column temperature of the chromatographic column during detection is 30-40 ℃. The chromatographic column is selected according to the effective components and impurities in the sample, and the type structure, polarity, acidity and alkalinity and molecular weight of the sample. The correct selection of the chromatographic column and the setting of the detection column temperature are beneficial to accurately and completely separating various substance components in the finished soup and displaying the substance components in a map.
When gradient elution is performed, elution may be performed in the following manner:
in 0-10 min, the volume proportion of acetonitrile in the mobile phase is increased from 2% to 6%, the volume proportion of water is reduced from 98% to 94%, and the flow rate of the mobile phase is 0.6-0.8 mL/min;
optionally, the flow rate of the mobile phase is 0.7 mL/min;
in 10-40 min, the volume proportion of acetonitrile in the mobile phase is increased from 6% to 15%, the volume proportion of water is reduced from 94% to 85%, and the flow rate of the mobile phase is 0.9-1.1 mL/min;
optionally, the flow rate of the mobile phase is 1.0 mL/min;
in 40-60 min, the volume ratio of acetonitrile in the mobile phase is 15%, the volume ratio of water is 85%, and the flow rate of the mobile phase is 0.9-1.1 mL/min;
optionally, the flow rate of the mobile phase is 1.0 mL/min;
in 60-80 min, the volume proportion of acetonitrile in the mobile phase is increased from 15% to 20%, the volume proportion of water is decreased from 85% to 80%, and the flow rate of the mobile phase is 0.9-1.1 mL/min;
optionally, the flow rate of the mobile phase is 1.0 mL/min;
in 80-100 min, the volume proportion of acetonitrile in the mobile phase is increased from 20% to 40%, the volume proportion of water is decreased from 80% to 60%, and the flow rate of the mobile phase is 0.9-1.1 mL/min;
alternatively, the flow rate of the mobile phase is 1.0 mL/min.
The application also provides a method for controlling the quality of the Wangban soup, which comprises the step of respectively taking an extracting solution, a concentrated solution, a dry paste and a finished product particle sample in the preparation process of the Wangban soup by adopting the method for establishing the Wangban soup fingerprint to detect fingerprints, wherein all the fingerprints at least comprise 13 common chromatographic peaks, and the similarity of all the fingerprints is more than or equal to 97 percent.
The method has the advantages that the number and the height of the common chromatographic peaks of the fingerprint of the extract, the concentrated solution, the dry paste and the finished product particle sample in the preparation process of the tape finishing soup and the similarity of the spectra are reflected by the difference of the spectra, so that the difference of various effective substance components of the tape finishing soup in the preparation process and in different batches is detected, and the quality of the tape finishing soup in the preparation process is controlled.
It should be noted that all fingerprint similarity degrees are obtained according to traditional Chinese medicine chromatogram fingerprint similarity degree evaluation system software (2012.130723 version), all fingerprint similarities needing to be compared are led into the traditional Chinese medicine chromatogram fingerprint similarity degree evaluation system software (2012.130723 version), HPLC fingerprints are established by a median method, a comparison characteristic spectrum formed by 13 common chromatographic peaks is generated, and the spectrum similarity degree is more than or equal to 97%.
The Wandai soup is prepared by the following steps:
1. raw material preparation
Weighing 30-40 parts of soil-fried bighead atractylodes rhizome, 30-40 parts of fried Chinese yam, 5-10 parts of ginseng, 15-20 parts of wine-fried white paeony root and 10-15 parts of wine-fried plantain seed according to parts by weight; 10-15 parts of rhizoma atractylodis; 3-5 parts of liquorice; 1-2 parts of dried orange peel, 1-2 parts of black schizonepeta spike and 2-3 parts of radix bupleuri.
The soil-fried bighead atractylodes rhizome is prepared by frying fine powder of ledum pith, hanging soil on the surface of the fine powder and screening excessive soil, wherein the mass ratio of bighead atractylodes rhizome tablets to the fine powder of ledum pith is 10: (1-3);
the semen plantaginis fried with wine is prepared by putting semen plantaginis into a frying container, heating with slow fire, frying until the semen plantaginis is slightly popped, spraying yellow wine, frying to dry, and cooling, wherein the mass ratio of semen plantaginis to yellow wine is 10: (0.08-0.12);
the black schizonepeta spike is prepared by cutting schizonepeta into segments, frying the schizonepeta into black brown by strong fire, storing the schizonepeta spike, spraying a small amount of clear water, taking out and drying the schizonepeta spike;
the processing method of other medicinal materials is described in 2015 edition of Chinese pharmacopoeia.
Optionally, weighing 37.5 parts of rhizoma atractylodis macrocephalae stir-fried with soil, 37.5 parts of rhizoma dioscoreae stir-fried with soil, 7.5 parts of ginseng, 18.75 parts of radix paeoniae alba stir-fried with wine and 11.25 parts of semen plantaginis stir-fried with wine according to parts by weight; 11.25 parts of prepared rhizoma atractylodis; 3.75 parts of liquorice; 1.875 parts of dried orange peel, 1.875 parts of black schizonepeta spike and 2.25 parts of radix bupleuri.
2. Preparing the Tanbaitang
Boiling the traditional Chinese medicine raw materials in water for 2-3 times, extracting for 0.5-2 h each time, wherein the water amount added each time is 6-8 times of the mass of the traditional Chinese medicine raw materials, combining extracting solutions, filtering by more than 100 meshes, centrifuging, concentrating under reduced pressure at 60-80 ℃ to obtain concentrated solution with the relative density of 1.05-1.20 (60-80 ℃), drying to obtain dry paste, wherein the paste yield is 20-30%, and adding auxiliary materials to prepare the finished product granular traditional Chinese medicine preparation.
The adjuvants include, but are not limited to, any one or more of maltodextrin, dextrin, powdered sugar, povidone, polyethylene glycol, xylitol, and beta-cyclodextrin.
Taking the extract sample as the above extract;
the concentrated solution sample is obtained by mixing 4g of concentrated solution with water, adopting a 50mL volumetric flask for constant volume, shaking up and filtering;
the dry paste sample is obtained by mixing 1g of dry paste with water, fixing the volume by adopting a 50mL volumetric flask, shaking up and filtering;
the finished product particle sample is obtained by mixing 1g of finished product particles with water, adopting a 50mL volumetric flask for constant volume, shaking up and filtering.
The concentration differences of the extract sample, the concentrated solution sample, the dry paste sample and the finished product particle sample are not large, and the subsequent characteristic map comparison is facilitated.
The fingerprint spectrum establishing method and the quality control method of the Wantangtang according to the present application will be further described in detail with reference to the following embodiments.
Example 1
The embodiment of the application provides a method for establishing a fingerprint of the Zodiac soup.
1. Preparing the Tanbaitang
Weighing 37.5 parts of soil-fried bighead atractylodes rhizome, 37.5 parts of fried Chinese yam, 7.5 parts of ginseng, 18.75 parts of wine-fried white paeony root and 11.25 parts of wine-fried plantain seed according to parts by weight; 11.25 parts of prepared rhizoma atractylodis; 3.75 parts of liquorice; boiling and extracting 1.875 parts of pericarpium citri reticulatae, 1.875 parts of black schizonepeta spike and 2.25 parts of radix bupleuri with water for 2 times, wherein the water addition amount is 6 times of the mass of the traditional Chinese medicinal materials each time, combining the extracting solutions, filtering, centrifuging, concentrating under reduced pressure at 70 ℃ to obtain a concentrated solution, drying to obtain a dry paste, adding auxiliary materials, and preparing to obtain a finished granular traditional Chinese medicine preparation;
2. preparation of a Dry paste sample
Mixing the prepared 1g of dry paste with water, fixing the volume by adopting a 50mL volumetric flask, shaking up, and filtering to obtain a dry paste sample;
3. establishing the characteristic fingerprint of Toanba Tang
In the embodiment of the application, 20 mu L of dry paste sample is absorbed for HPLC detection, and the measurement wavelength of an ultraviolet detector is set to be 240-250 nm; selecting a C18 water-resistant chromatographic column as a chromatographic column; the temperature of the chromatographic column is 30-40 ℃ during detection; acetonitrile and water were used as mobile phases, which were eluted in the following manner, as shown in table 1:
table 1 example 1HPLC determination of mobile phase and flow rate
The obtained HPLC fingerprint is shown in FIG. 1, wherein the peaks of various substance components are obvious, and the separation degree and the signal-to-noise ratio are high.
Example 2
The embodiment of the application provides a method for controlling the quality of a tao-chun soup.
1. Preparation of samples
Preparing 10 batches of the tape-out soup according to the method of example 1, and taking 10 batches of dry paste to prepare dry paste samples according to the method of example 1;
2. establishing quality control of Toanba decoction
HPLC fingerprints of 10 batches of dry paste samples of the soup with complete strip are respectively measured according to the HPLC detection method of example 1, all the fingerprints are introduced into traditional Chinese medicine chromatographic fingerprint similarity evaluation system software (2012.130723 version), and the HPLC fingerprint is established by a median method to generate a control characteristic spectrum consisting of 13 common chromatographic peaks, as shown in figure 2.
Example 3
The embodiment of the application provides a method for controlling the quality of a tao-chun soup.
1. Preparation of samples
The extract, the concentrate, the dry paste and the finished granules of the tape finishing soup prepared in example 1 were respectively used to prepare samples according to the following methods:
taking the extract sample as the above extract;
the concentrated solution sample is obtained by mixing 4g of concentrated solution with water, adopting a 50mL volumetric flask for constant volume, shaking up and filtering;
the dry paste sample is obtained by mixing 1g of dry paste with water, fixing the volume by adopting a 50mL volumetric flask, shaking up and filtering;
the finished product particle sample is prepared by mixing 1g of finished product particles with water, adopting a 50mL volumetric flask for constant volume, shaking up and filtering;
2. establishing quality control of Toanba decoction
HPLC fingerprints of the extract sample, the concentrated solution sample, the dry paste sample and the finished product granule sample are respectively measured according to the HPLC detection method of example 1, as shown in FIG. 3, corresponding positions of each fingerprint have the same 13 chromatographic peaks, and the similarity is calculated and is shown in Table 2:
TABLE 2 fingerprint similarity calculation Table
As can be seen from Table 2, the similarity of each chromatogram was not less than 97%.
Comparative example 1
The application provides a method for establishing the fingerprint of the Zongtang.
Respectively taking the dry paste samples prepared in the embodiment 1 to carry out HPLC detection, and setting the measurement wavelength of an ultraviolet detector to be 240-250 nm; selecting a C18 water-resistant chromatographic column as a chromatographic column; the temperature of the chromatographic column is 30-40 ℃ during detection; acetonitrile and water were used as mobile phases, which were eluted in the following manner, as shown in table 3:
table 3 example 1HPLC determination of mobile phase and flow rate
The obtained HPLC fingerprint is shown in FIG. 1.
Comparative example 2
The application provides a method for establishing the fingerprint of the Zongtang.
Respectively taking the dry paste samples prepared in the embodiment 1 to carry out HPLC detection, and setting the measurement wavelength of an ultraviolet detector to be 240-250 nm; selecting a C18 water-resistant chromatographic column as a chromatographic column; the temperature of the chromatographic column is 30-40 ℃ during detection; acetonitrile and water were used as mobile phases, which were eluted in the following manner, as shown in table 4:
table 4 example 1HPLC determination of mobile phase and flow rate
The obtained HPLC fingerprint is shown in FIG. 1.
Comparing the three spectra in FIG. 1, the spectra obtained in example 1 have obvious peak separation of various substance components and high separation degree and signal-to-noise ratio; the spectrum obtained in comparative example 1, due to poor sample separation, shows almost all substances coming out of the chromatographic column together and does not well show the characteristic peak of each substance component; although the spectrum obtained in comparative example 2 is similar to that of the example, the separation degree and the signal-to-noise ratio are not good as in example 1, and the chromatographic column is damaged greatly by eluting with pure water in comparative example 2.
In summary, according to the method for establishing the finger print of the tape finishing soup provided by the embodiment of the application, the finger print of the tape finishing soup sample is detected by a high performance liquid chromatograph, the tape finishing soup is prepared from raw materials including rhizoma atractylodis macrocephalae, rhizoma dioscoreae, ginseng, radix paeoniae alba, semen plantaginis, rhizoma atractylodis, liquorice, pericarpium citri reticulatae, black schizonepeta spike and radix bupleuri, and the tape finishing soup contains various effective substance components. According to the method, acetonitrile and water are used as mobile phases to carry out gradient elution on the soup sample, so that various substance components in the soup sample can be accurately separated, and the complete fingerprint with an obvious characteristic peak pattern is obtained.
A quality control method of the tape finishing soup comprises the steps of comparing fingerprint spectrums of extracting solution, concentrated solution, dry paste and finished product particle samples of different batches of tape finishing soup in the preparation process, and comparing the number and the height of common chromatographic peaks to determine whether various effective substance components in the extracting solution, the concentrated solution, the dry paste and the finished product particle samples of the tape finishing soup in the preparation process are the same or not, and whether various effective substance components and different batches of the tape finishing soup in the preparation process and different batches of the tape finishing soup are changed or not, so that the quality of the tape finishing soup in the preparation process is controlled.
The foregoing is illustrative of the present application and is not to be construed as limiting thereof, as numerous modifications and variations will be apparent to those skilled in the art. Any modification, equivalent replacement, improvement and the like made within the spirit and principle of the present application shall be included in the protection scope of the present application.
Claims (8)
1. The method for establishing the fingerprint of the Zodiac soup is characterized by comprising the steps of detecting the fingerprint of a sample of the Zodiac soup by using a high performance liquid chromatograph;
during detection, acetonitrile and water are selected as mobile phases, gradient elution is adopted, and the flow rate of the mobile phases is 0.6-1.1 mL/min;
the volume proportion of the acetonitrile in the mobile phase is 2-40%;
when the gradient elution is carried out, the following elution is adopted:
in 0-10 min, the volume proportion of acetonitrile in the mobile phase is increased from 2% to 6%;
in 10-40 min, the volume proportion of acetonitrile in the mobile phase is increased from 6% to 15%;
in 40-60 min, the volume ratio of acetonitrile in the mobile phase is 15%;
increasing the volume proportion of acetonitrile in the mobile phase from 15% to 20% in 60-80 min;
in 80-100 min, the volume proportion of acetonitrile in the mobile phase is increased from 20% to 40%;
the chromatographic column used for detection is a C18 water-resistant chromatographic column;
the TAO TANG is prepared from Atractylodis rhizoma, rhizoma Dioscoreae, Ginseng radix, radix Paeoniae alba, semen plantaginis, rhizoma Atractylodis, Glycyrrhrizae radix, pericarpium Citri Tangerinae, herba Schizonepetae and bupleuri radix.
2. The method for establishing fingerprint of TAI TANG according to claim 1, wherein the temperature of the chromatographic column is 30-40 ℃ during detection.
3. The method for establishing a fingerprint of a quan zhi tang according to claim 1, wherein the detection wavelength of the ultraviolet detector is set to 240 to 250nm when the liquid phase detection is performed.
4. The method for establishing a fingerprint of a Wanyi decoction according to claim 1, wherein the gradient elution is performed at a flow rate of 0.6-0.8 mL/min for 0-10 min and at a flow rate of 0.9-1.1 mL/min for 10-100 min.
5. The method for establishing a fingerprint of a Wanyi decoction according to claim 4, wherein the gradient elution is performed at a flow rate of 0.7mL/min to 10min and at a flow rate of 1.0mL/min to 100 min.
6. A quality control method of the Tou Belt decoction is characterized in that extract samples, concentrated solution samples, dry paste samples and finished product particle samples of different batches of Tou Belt decoction in the preparation process are respectively taken to detect fingerprint spectrums by adopting the fingerprint spectrum establishment method of the Tou Belt decoction according to any one of claims 1 to 5, all the fingerprint spectrums at least comprise 13 common chromatographic peaks, and the similarity of all the fingerprint spectrums is more than or equal to 97 percent.
7. The method for controlling quality of Wangbai Tang according to claim 6, wherein the Wangbai Tang is prepared by:
according to parts by weight, 30-40 parts of soil-fried bighead atractylodes rhizome, 30-40 parts of fried Chinese yam, 5-10 parts of ginseng, 15-20 parts of wine-fried white paeony root and 10-15 parts of wine-fried plantain seed are added; 10-15 parts of rhizoma atractylodis; 3-5 parts of liquorice; adding water into 1-2 parts of pericarpium citri reticulatae, 1-2 parts of black schizonepeta spike and 2-3 parts of radix bupleuri, heating and extracting for 2-3 times, each time for 0.5-2 hours, combining extracting solutions, filtering, concentrating to obtain a concentrated solution, drying to obtain a dry paste, and adding auxiliary materials to prepare a finished granular traditional Chinese medicine preparation.
8. The quality control method of the Wanyi decoction according to claim 6, wherein the extract solution sample, the concentrated solution sample, the dry paste sample and the finished pellet sample are obtained by:
filtering the extracting solution to obtain the extracting solution sample;
mixing 4g of the concentrated solution with water, fixing the volume to 50mL, and filtering to obtain a concentrated solution sample;
mixing 1g of the dry paste with water, diluting to 50mL, and filtering to obtain a dry paste sample;
and (3) mixing 1g of the finished product particles with water, metering the volume to 50mL, and filtering to obtain the finished product particle sample.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910772761.1A CN110455973B (en) | 2019-08-21 | 2019-08-21 | Method for establishing fingerprint spectrum of Tou Tang and method for controlling quality of Tou Tang |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910772761.1A CN110455973B (en) | 2019-08-21 | 2019-08-21 | Method for establishing fingerprint spectrum of Tou Tang and method for controlling quality of Tou Tang |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110455973A CN110455973A (en) | 2019-11-15 |
| CN110455973B true CN110455973B (en) | 2021-08-17 |
Family
ID=68488172
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910772761.1A Active CN110455973B (en) | 2019-08-21 | 2019-08-21 | Method for establishing fingerprint spectrum of Tou Tang and method for controlling quality of Tou Tang |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110455973B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114563510B (en) * | 2022-03-01 | 2022-10-14 | 山东大学 | Establishing method of fingerprint spectrum of minor radix bupleuri decoction formula based on shaoyang disease formula |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1248721C (en) * | 2003-05-19 | 2006-04-05 | 广东明珠药业有限公司 | Chinese medicine composition for treating consumptive disease and its quality control method |
| CN107064320B (en) * | 2016-10-21 | 2019-09-24 | 广东省中医院 | A kind of method of quality control and its finger-print of reinforcing spleen and kidney side |
| CN109828069A (en) * | 2019-03-15 | 2019-05-31 | 合肥创新医药技术有限公司 | A kind of measuring method of Chinese medicine compound prescription yihuang decoction finger-print |
| CN109917048B (en) * | 2019-04-18 | 2022-03-08 | 江阴天江药业有限公司 | Construction method and application of UPLC characteristic spectrum of cortex fraxini medicinal material |
-
2019
- 2019-08-21 CN CN201910772761.1A patent/CN110455973B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN110455973A (en) | 2019-11-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107153098A (en) | A kind of assay method of the finger-print of Chinese medicine composition | |
| CN105486771B (en) | The fingerprint atlas detection method of Xiao Chai Hu granules compound preparation | |
| CN101062294B (en) | Method for detecting the quality of the medicine combination for treating cerebral apoplexy and vasculitis | |
| CN105588885A (en) | Salvianolic acid extract fingerprint spectrum and content measurement method of related components | |
| CN112697948B (en) | Quality detection method of lung-clearing and toxin-expelling soup established based on fingerprint model | |
| CN106645535A (en) | Method for measuring content of five saponin components in red ginseng roots | |
| CN110455973B (en) | Method for establishing fingerprint spectrum of Tou Tang and method for controlling quality of Tou Tang | |
| CN108459090B (en) | Quality control method of Jichuan decoction composition | |
| CN107340348A (en) | A kind of method for building up of Bletilla striata medicinal materials HPLC finger-prints | |
| CN108355041A (en) | A kind of preparation process of Gualou Xiebai Banxia Tang solid pharmaceutical preparation | |
| CN103115984A (en) | Quality control method of medicament for treating leukopenia | |
| CN103344738B (en) | Detection method of nine-component heart-calming particle | |
| CN100558386C (en) | A kind of ginseng glucose injection | |
| CN106918673B (en) | A kind of method for building up of the finger-print of Chinese medicine composition | |
| CN112666277A (en) | Method for constructing and detecting HPLC (high performance liquid chromatography) characteristic spectrum of rhizoma cyperi medicinal materials, decoction pieces, standard decoction and formula granules | |
| CN108982738B (en) | Construction method and detection method of intermediate characteristic spectrum of Zhenqi Fuzheng granule or Zhenqi Fuzheng granule fingerprint | |
| CN108061762A (en) | The content assaying method of three kinds of flavone components in a kind of kidney tonifying eliminating toxic particle | |
| CN112666281B (en) | Production method of lung clearing and toxin expelling soup established based on fingerprint model | |
| CN115078608B (en) | Method for identifying UPLC characteristic patterns of herba abri and herba abri | |
| CN115541756A (en) | Fingerprint spectrum construction method, detection method and identification method of plantago Chinese medicinal material or preparation | |
| CN106526000B (en) | The method for building up and finger-print of Shengma Gegen Tang composition finger-print | |
| CN103463211A (en) | Sugar-free prepared multiflower knotweed tuber granules, preparation method and quality control method thereof | |
| CN107389827A (en) | The assay method of stachydrine hydrochloride content in motherwort or its compound preparation | |
| CN103592390A (en) | Quality detection method of chlorogenic acid in Yinmu liver-relieving granules | |
| CN107607653A (en) | The method for determining Radix Scrophulariae extract finger-print |
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 | ||
| CP03 | Change of name, title or address | ||
| CP03 | Change of name, title or address |
Address after: 435000 9 Shengming Road, Jinshan street, Huangshi economic and Technological Development Zone, Hubei Province Patentee after: Jingpai Zhengtang Pharmaceutical Co.,Ltd. Address before: No.9, Shengming Road, Jinshan street, Huangshi economic and Technological Development Zone, Huangshi City, Hubei Province Patentee before: JING BRAND BIO-MEDICINE Co.,Ltd. |