CN114113435A - Method for constructing characteristic spectrum of dried orange peel medicinal material and preparation - Google Patents
Method for constructing characteristic spectrum of dried orange peel medicinal material and preparation Download PDFInfo
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Abstract
The invention relates to the technical field of quality analysis and control of medicinal materials and preparations thereof, in particular to a method for constructing a characteristic map of a dried orange peel medicinal material and a preparation thereof. The construction method of the characteristic map of the dried orange peel medicinal material and the preparation comprises the following steps: detecting the test solution by using an HPLC method to obtain a characteristic spectrum; the detection conditions of the HPLC method include: and acetonitrile is used as a mobile phase A, and water is used as a mobile phase B for gradient elution. The invention establishes the determination method of the characteristic spectrum of the dried orange peel medicinal material and the preparation, and realizes the good separation of chromatographic peaks; according to the construction method of the characteristic spectrum of the dried orange peel medicinal material and the preparation, the obtained characteristic spectrum contains 8 characteristic peaks of the dried orange peel medicinal material and 7 characteristic peaks of a dried orange peel preparation such as dried orange peel formula granules, so that the main components of the dried orange peel medicinal material or the preparation can be comprehensively represented, and the method has very important significance for comprehensively evaluating the quality of the dried orange peel medicinal material or the preparation.
Description
Technical Field
The invention relates to the technical field of quality analysis and control of medicinal materials and preparations thereof, in particular to a method for constructing a characteristic map of a dried orange peel medicinal material and a preparation thereof.
Background
The pericarpium Citri Tangerinae is dried mature pericarp of Rutaceae plant Citrus reticulata Blanco and its cultivar. Has effects of regulating qi-flowing, lowering adverse qi, regulating middle warmer, stimulating appetite, eliminating dampness, and eliminating phlegm. At present, the dried orange peel is only used for controlling the content of hesperidin in the 2020 edition of pharmacopoeia of the people's republic of China, an HPLC characteristic spectrum is not established for medicinal materials, and the aim of comprehensively evaluating the quality of the medicinal materials and preparations thereof cannot be fulfilled.
The characteristic spectrum of the traditional Chinese medicine refers to the spectrum or chromatogram of a certain kind or several kinds of components which are common in a certain traditional Chinese medicine and a preparation thereof and have characteristics, and the establishment of the characteristic spectrum has very important significance for effectively controlling the quality of the medicinal material and the preparation thereof.
In view of the above, the present invention is particularly proposed.
Disclosure of Invention
The invention aims to provide a method for constructing a characteristic spectrum of a dried orange peel medicinal material and a preparation, and aims to solve the technical problem that the quality of the dried orange peel medicinal material and the preparation can not be comprehensively evaluated in the prior art.
In order to achieve the above purpose of the present invention, the following technical solutions are adopted:
the construction method of the characteristic map of the dried orange peel medicinal material and the preparation comprises the following steps:
detecting the test solution by using an HPLC method to obtain a characteristic spectrum;
the detection conditions of the HPLC method include: taking acetonitrile as a mobile phase A and water as a mobile phase B for gradient elution; the gradient elution comprises: changing the volume fraction of the mobile phase A from 21-23% to 24-26% in 0-15 min; changing the volume fraction of the mobile phase A from 24-26% to 29-31% in 15-25 min; changing the volume fraction of the mobile phase A from 29-31% to 49-51% in 25-30 min; changing the volume fraction of the mobile phase A from 49-51% to 59-61% in 30-35 min; changing the volume fraction of the mobile phase A from 59-61% to 62-63% in 35-45 min; and (4) changing the volume fraction of the mobile phase A from 62-63% to 67-69% in 45-50 min.
The method adopts a certain detection method, chromatographic conditions and the like to establish and obtain the characteristic spectrum of the dried orange peel medicinal material, integrates 0.5 percent of the area of the hesperidin to obtain 8 characteristic peaks in the characteristic spectrum of the dried orange peel medicinal material, integrates 3.0 percent of the area of the dried orange peel preparation such as dried orange peel formula granules to obtain 7 characteristic peaks in the dried orange peel formula granules, and can evaluate the quality of the dried orange peel medicinal material through the overall surface of various components so as to realize the quality control of the dried orange peel medicinal material and the preparation.
In a specific embodiment of the present invention, the method further comprises: detecting the reference substance solution by adopting the HPLC method; the reference substance comprises one or more of narirutin, hesperidin, nobiletin and hesperetin. Further, the control comprises narirutin, hesperidin, nobiletin and hesperetin.
In a specific embodiment of the present invention, in the HPLC method, the column uses octadecylsilane bonded silica as a packing material. Further, the column may be a 5 μm, 4.6 × 250mm column.
In a specific embodiment of the invention, the detection wavelength in the HPLC method is 280-290 nm.
In a specific embodiment of the invention, in the HPLC method, the flow rate of the mobile phase is 0.8-1.2 mL/min.
In a specific embodiment of the invention, in the HPLC method, the column temperature of the chromatographic column is 25-35 ℃.
In a specific embodiment of the invention, the gradient elution is:
in a particular embodiment of the invention, the number of theoretical plates should not be lower than 2000 calculated on the hesperidin peak.
In a specific embodiment of the present invention, the sample volume of the sample solution is 5 to 10 μ L.
In a specific embodiment of the present invention, the method for preparing the test solution comprises:
when the substance to be detected is a dried orange peel medicinal material, a solid or semisolid dried orange peel preparation, performing ultrasonic extraction on the dried orange peel medicinal material or the dried orange peel preparation by using methanol as an extraction solvent, supplementing the loss weight by using the extraction solvent, filtering, and taking a subsequent filtrate as a test solution;
when the substance to be detected is a liquid dried orange peel preparation, the liquid can be directly taken as a test solution for analysis.
In a specific embodiment of the invention, the preparation is a dried orange peel single-flavor preparation. Furthermore, the pericarpium citri reticulatae single-ingredient preparation is pericarpium citri reticulatae formula granules.
In a specific embodiment of the invention, when the substance to be detected is a dried orange peel drug, the ratio of the dried orange peel drug to the extraction solvent is (0.1-0.3) g: 25mL, such as 0.2 g: 25 mL.
In a specific embodiment of the invention, when the dried orange peel preparation is in a solid or semisolid dosage form of the substance to be tested, the ratio of the dried orange peel preparation to the extraction solvent is (0.1-0.3) g: 50mL, such as 0.2 g: 50 mL.
In a specific embodiment of the invention, the ultrasonic extraction time is 30-60 min; the power of the ultrasonic wave is 250-350W, and the frequency of the ultrasonic wave is 35-45 kHz.
In actual operation, dried orange peel crude powder is adopted for ultrasonic extraction. Further, the coarse powder is powder passing through a second sieve.
In a specific embodiment of the present invention, the method for preparing the control solution comprises: precisely weighing narirutin, hesperidin, nobiletin and hesperetin, and dissolving with methanol.
The invention compares the characteristic peaks in the characteristic map with a reference substance and identifies the chemical components corresponding to 4 characteristic peaks in the characteristic map.
In the specific implementation mode of the invention, more than 10 batches of characteristic maps of the pericarpium citri reticulatae medicinal materials or the preparations are constructed according to the HPLC method, and a traditional Chinese medicine chromatogram fingerprint similarity evaluation system (2.0 version) is adopted to synthesize the control maps of the pericarpium citri reticulatae medicinal materials or the preparations.
In a specific embodiment of the invention, the control map of the pericarpium citri reticulatae medicinal material has 8 characteristic peaks, wherein the peak 1#, the peak 2#, the peak 6# and the peak 8# respectively correspond to peaks of a narirutin control, a hesperidin control, a nobiletin control and an hesperetin control, the peak 2# is taken as a reference peak S, relative retention times of the peak 3#, the peak 4#, the peak 5# and the peak 7# and the reference peak S are calculated, and relative peak areas of the peak 1#, the peak 6# and the peak 8# and the reference peak S are calculated as follows:
the relative retention time of the peak 3 is 2.10-2.18; the relative retention time of the 4# peak is 2.95-3.07; the relative retention time of the 5# peak is 3.10-3.22; the relative retention time of the 7# peak is 3.32-3.46;
1#the relative peak area of the peak is not lower than 0.013; the relative peak area of the No. 6 peak is not lower than 0.022; the relative peak area of the 8# peak was not less than 0.007.
In a specific embodiment of the present invention, the control map of the tangerine peel formula granule has 7 characteristic peaks, wherein the peak 1#, the peak 2#, the peak 5# and the peak 7# respectively correspond to the peaks of a narirutin control, a hesperidin control, a nobiletin control and an hesperetin control, the peak 2# is used as a reference peak S, the relative retention time of the peak 3#, the peak 4# and the peak 6# and the reference peak S is calculated, and the relative peak areas of the peak 1#, the peak 5# and the peak 7# and the reference peak S are respectively as follows:
3#the relative retention time of the peak is 2.95-3.07; 4#The relative retention time of the peaks is 3.10-3.22; 6#The relative retention time of the peaks is 3.32-3.46;
1#the relative peak area of the peak is not lower than 0.06; 5#The relative peak area of the peak is not lower than 0.23; 7#The relative peak area of the peak is not less than 0.05.
In actual operation, the characteristic spectrum of a sample to be detected can be introduced into traditional Chinese medicine fingerprint spectrum similarity evaluation software, the similarity between the characteristic spectrum and a reference spectrum is calculated, and the quality control of the dried orange peel medicinal material or the preparation is carried out through the similarity.
Compared with the prior art, the invention has the beneficial effects that:
(1) the invention establishes the determination method of the characteristic spectrum of the dried orange peel medicinal material and the preparation, and realizes the good separation of chromatographic peaks; according to the construction method of the characteristic spectrum of the dried orange peel medicinal material and the preparation, the obtained characteristic spectrum contains 8 characteristic peaks of the dried orange peel medicinal material and 7 characteristic peaks of a dried orange peel preparation such as dried orange peel formula granules, so that the main components of the dried orange peel medicinal material or the preparation can be comprehensively represented, and the method has very important significance for comprehensively evaluating the quality of the dried orange peel medicinal material or the preparation;
(2) the invention provides a standard control map of dried orange peel medicinal materials and preparations, which can be used for analyzing and determining the relative contents of main components in the dried orange peel medicinal materials and the preparations, and can effectively and reliably predict the product quality through the contents.
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 a dried orange peel medicinal material provided in embodiment 1 of the present invention;
fig. 2 is a characteristic spectrum of the tangerine peel formula particle provided in example 2 of the present invention;
fig. 3 is a feature spectrum of 10 batches of pericarpium citri reticulatae medicinal materials provided in embodiment 3 of the present invention;
FIG. 4 is a comparison feature map of the synthesis of 10 batches of pericarpium Citri Tangerinae drugs provided in example 3 of the present invention;
fig. 5 is a feature map of a 10-batch tangerine peel formula particle provided in example 4 of the present invention;
FIG. 6 is a comparison feature map of the synthesis of 10 batches of tangerine peel granules provided in example 4 of the present invention;
fig. 7 is a characteristic map of the dried orange peel medicinal material provided in comparative example 1.
Detailed Description
The technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings and the detailed description, but those skilled in the art will understand that the following described embodiments are some, not all, of the embodiments of the present invention, and are only used for illustrating the present invention, and should not be construed as limiting the scope of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention. 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.
The construction method of the characteristic map of the dried orange peel medicinal material and the preparation comprises the following steps:
detecting the test solution by using an HPLC method to obtain a characteristic spectrum;
the detection conditions of the HPLC method include: taking acetonitrile as a mobile phase A and water as a mobile phase B for gradient elution; the gradient elution comprises: changing the volume fraction of the mobile phase A from 21-23% to 24-26% in 0-15 min; changing the volume fraction of the mobile phase A from 24-26% to 29-31% in 15-25 min; changing the volume fraction of the mobile phase A from 29-31% to 49-51% in 25-30 min; changing the volume fraction of the mobile phase A from 49-51% to 59-61% in 30-35 min; changing the volume fraction of the mobile phase A from 59-61% to 62-63% in 35-45 min; and (4) changing the volume fraction of the mobile phase A from 62-63% to 67-69% in 45-50 min.
The method adopts a certain detection method, chromatographic conditions and the like to establish and obtain the characteristic spectrum of the dried orange peel medicinal material, integrates 0.5 percent of the area of the hesperidin to obtain 8 characteristic peaks in the characteristic spectrum of the dried orange peel medicinal material, integrates 3.0 percent of the area of the dried orange peel preparation such as dried orange peel formula granules to obtain 7 characteristic peaks in the dried orange peel formula granules, and can evaluate the quality of the dried orange peel medicinal material through the overall surface of various components so as to realize the quality control of the dried orange peel medicinal material and the preparation.
In a specific embodiment of the present invention, the method further comprises: detecting the reference substance solution by adopting the HPLC method; the reference substance comprises one or more of narirutin, hesperidin, nobiletin and hesperetin. Further, the control comprises narirutin, hesperidin, nobiletin and hesperetin.
In a specific embodiment of the present invention, in the HPLC method, the column uses octadecylsilane bonded silica as a packing material. Further, the column may be a 5 μm, 4.6 × 250mm column.
In a specific embodiment of the invention, in the HPLC method, the detection wavelength is 280-290 nm, preferably 280-285 nm, and more preferably 283 nm.
As in the different embodiments, the detection wavelength may be 280nm, 281nm, 282nm, 283nm, 284nm, 285nm, 286nm, 287nm, 288nm, 289nm, 290nm, and so forth.
In a specific embodiment of the present invention, the flow rate of the mobile phase in the HPLC method is 0.8-1.2 mL/min, preferably 0.9-1.1 mL/min, and more preferably 1 mL/min.
In a specific embodiment of the present invention, in the HPLC method, the column temperature of the chromatographic column is 25 to 35 ℃, preferably 28 to 32 ℃, and more preferably 30 ℃.
As in the different embodiments, the column temperature of the chromatographic column can be 25 ℃, 26 ℃, 27 ℃, 28 ℃, 29 ℃, 30 ℃, 31 ℃, 32 ℃, 33 ℃, 34 ℃, 35 ℃ and so on.
In a specific embodiment of the invention, the gradient elution is:
| time: min | Mobile phase A: is based on | Mobile phase B: is based on |
| 0～15 | 22→25 | 78→75 |
| 15～25 | 25→30 | 75→70 |
| 25～30 | 30→50 | 70→50 |
| 30～35 | 50→60 | 50→40 |
| 35～45 | 60→62 | 40→38 |
| 45～50 | 62→68 | 38→32 |
。
In a particular embodiment of the invention, the number of theoretical plates should not be lower than 2000 calculated on the hesperidin peak.
In a specific embodiment of the present invention, the sample volume of the sample solution is 5 to 10 μ L.
In a specific embodiment of the present invention, the detection conditions of the HPLC method include:
octadecylsilane chemically bonded silica is used as a filler for the chromatographic column; the column specification is 5 μm, 4.6 × 250 mm; taking acetonitrile as a mobile phase A and water as a mobile phase B for gradient elution;
the detection wavelength is 283 nm; the flow rate is 1 mL/min; the column temperature is 30 ℃; the sample injection amount is 5-10 mu L; the number of theoretical plates is not less than 2000 calculated according to the hesperidin peak;
the gradient elution is:
| time: min | Mobile phase A: is based on | Mobile phase B: is based on |
| 0～15 | 22→25 | 78→75 |
| 15～25 | 25→30 | 75→70 |
| 25～30 | 30→50 | 70→50 |
| 30～35 | 50→60 | 50→40 |
| 35～45 | 60→62 | 40→38 |
| 45～50 | 62→68 | 38→32 |
。
In a specific embodiment of the present invention, the method for preparing the test solution comprises:
when the substance to be detected is a dried orange peel medicinal material, a solid or semisolid dried orange peel preparation, performing ultrasonic extraction on the dried orange peel medicinal material or the dried orange peel preparation by using methanol as an extraction solvent, supplementing the loss weight by using the extraction solvent, filtering, and taking a subsequent filtrate as a test solution;
when the substance to be detected is a liquid dried orange peel preparation, the liquid can be directly taken as a test solution for analysis.
In a specific embodiment of the invention, the preparation is a dried orange peel single-flavor preparation. Furthermore, the pericarpium citri reticulatae single-ingredient preparation is pericarpium citri reticulatae formula granules.
In a specific embodiment of the invention, when the substance to be detected is a dried orange peel drug, the ratio of the dried orange peel drug to the extraction solvent is (0.1-0.3) g: 25mL, such as 0.2 g: 25 mL.
In various embodiments, the ratio of dried citrus peel material to extraction solvent can be 0.1 g/25 mL, 0.15 g/25 mL, 0.2 g/25 mL, 0.25 g/25 mL, 0.3 g/25 mL, and the like.
In a specific embodiment of the invention, when the dried orange peel preparation is in a solid or semisolid dosage form of the substance to be tested, the ratio of the dried orange peel preparation to the extraction solvent is (0.1-0.3) g: 50mL, such as 0.2 g: 50 mL.
In various embodiments, the ratio of the preparation to the extraction solvent can be 0.1 g: 50mL, 0.15 g: 50mL, 0.2 g: 50mL, 0.25 g: 50mL, 0.3 g: 50mL, and the like.
In a specific embodiment of the invention, the ultrasonic extraction time is 30-60 min; the power of the ultrasonic wave is 250-350W, and the frequency of the ultrasonic wave is 35-45 kHz.
As in different embodiments, the time of the ultrasonic extraction may be 30min, 35min, 40min, 45min, 50min, 55min, 60min, and so on; in actual operation, the ultrasonic extraction time can be properly prolonged for the dried orange peel medicinal material, for example, 40-50 min; for the dried orange peel preparation, such as dried orange peel formula granules, the ultrasonic extraction time can be properly shortened, such as 30-35 min.
In actual operation, dried orange peel crude powder is adopted for ultrasonic extraction. Further, the coarse powder is powder passing through a second sieve.
In a specific embodiment of the present invention, the method for preparing the control solution comprises: precisely weighing narirutin, hesperidin, nobiletin and hesperetin, and dissolving with methanol. Further, in the control solution, the concentrations of the narirutin, the hesperidin, the nobiletin and the hesperetin are 100 mug/mL, 100 mug/mL and 100 mug/mL respectively.
The invention compares the characteristic peaks in the characteristic map with a reference substance, identifies the chemical components corresponding to 4 characteristic peaks in the characteristic map, and the identified 4 characteristic peaks are narirutin, hesperidin, nobiletin and hesperetin respectively.
In the specific implementation mode of the invention, more than 10 batches of characteristic maps of the pericarpium citri reticulatae medicinal materials or the preparations are constructed according to the HPLC method, and a traditional Chinese medicine chromatogram fingerprint similarity evaluation system (2.0 version) is adopted to synthesize the control maps of the pericarpium citri reticulatae medicinal materials or the preparations.
In a specific embodiment of the invention, the control map of the dried orange peel medicinal material has 8 characteristic peaks, wherein the peak 1#, the peak 2#, the peak 6# and the peak 8# respectively correspond to the peaks of a narirutin control, a hesperidin control, a nobiletin control and an hesperetin control, and the peaks are respectively expressed by 2#The peak is a reference peak S, the relative retention time of the 3# peak, the 4# peak, the 5# peak and the 7# peak with the reference peak S is calculated, and the relative peak areas of the 1# peak, the 6# peak and the 8# peak with the reference peak S are calculated, respectively as follows:
the relative retention time of the peak 3 is 2.10-2.18; the relative retention time of the 4# peak is 2.95-3.07; the relative retention time of the 5# peak is 3.10-3.22; the relative retention time of the 7# peak is 3.32-3.46;
1#the relative peak area of the peak is not lower than 0.013; the relative peak area of the No. 6 peak is not lower than 0.022; the relative peak area of the 8# peak was not less than 0.007.
In a specific embodiment of the invention, the control spectrum of the tangerine peel formula particle has 7 characteristic peaks, wherein the peak 1#, the peak 2#, the peak 5#, and the peak 7# respectively correspond to the peaks of a narirutin control, a hesperidin control, a nobiletin control, and an hesperetin control, and are respectively expressed by 2#The peak is a reference peak S, the relative retention time of the 3# peak, the 4# peak and the 6# peak with the reference peak S is calculated, and the relative peak areas of the 1# peak, the 5# peak and the 7# peak with the reference peak S are calculated, respectively as follows:
3#the relative retention time of the peak is 2.95-3.07; 4#The relative retention time of the peaks is 3.10-3.22; 6#The relative retention time of the peaks is 3.32-3.46;
1#the relative peak area of the peak is not lower than 0.06; 5#The relative peak area of the peak is not lower than 0.23; 7#The relative peak area of the peak is not less than 0.05.
In actual operation, the characteristic spectrum of a sample to be detected can be introduced into traditional Chinese medicine fingerprint spectrum similarity evaluation software, the similarity between the characteristic spectrum and a reference spectrum is calculated, and the quality control of the dried orange peel medicinal material or the preparation is carried out through the similarity.
In actual operation, the quality of the pericarpium citri reticulatae medicinal material or the preparation is evaluated according to the numerical value of the similarity, and the numerical boundary of the similarity can be defined according to actual requirements so as to judge whether the quality of the pericarpium citri reticulatae medicinal material or the preparation is qualified.
Example 1
The embodiment provides a method for constructing a characteristic spectrum of a dried orange peel medicinal material, which comprises the following steps:
(1) preparation of test solution
Precisely weighing 0.2g of coarse powder of dried orange peel medicinal materials (sieved by a second sieve), placing the coarse powder in a conical flask with a plug, precisely adding 25mL of methanol, sealing the plug, weighing, carrying out ultrasonic treatment (power 300W and frequency 40kHz) for 45min, cooling, weighing again, complementing the loss weight with methanol, shaking up, filtering, and taking a subsequent filtrate to obtain a test solution.
(2) Preparation of control solutions
Precisely weighing appropriate amount of narirutin control, hesperidin control, nobiletin control and hesperetin control, and adding methanol to obtain mixed solution containing 100 μ g of control per 1mL to obtain control solution.
(3) Detection conditions
The chromatographic column takes octadecylsilane chemically bonded silica as a filler (Phenomenex Gemini, 5 mu m, 4.6 multiplied by 250 mm); taking acetonitrile as a mobile phase A and water as a mobile phase B, and carrying out gradient elution, wherein the specific gradient elution procedure is shown in table 1; detection wavelength: 283 nm; flow rate: 1 mL/min; column temperature: 30 ℃; sample introduction amount: 10 mu L of the solution; the number of theoretical plates is not less than 2000 calculated according to hesperidin peak.
TABLE 1 gradient elution procedure (volume fraction)
| Time: min | Mobile phase A: is based on | Mobile phase B: is based on |
| 0～15 | 22→25 | 78→75 |
| 15～25 | 25→30 | 75→70 |
| 25～30 | 30→50 | 70→50 |
| 30～35 | 50→60 | 50→40 |
| 35～45 | 60→62 | 40→38 |
| 45～50 | 62→68 | 38→32 |
(4) Detection step
Precisely absorbing 10 μ L of reference solution and sample solution, respectively, injecting into high performance liquid chromatograph for determination, and recording chromatogram, as shown in FIG. 1, which is the characteristic spectrum (two batches) of pericarpium Citri Tangerinae. Wherein 1 is#Peak corresponds to narirutin, 2#The peak is a reference peak corresponding to hesperidin, 6#Peak corresponds to nobiletin, 8#The peak corresponds to hesperetin.
Example 2
The embodiment provides a method for constructing a characteristic spectrum of tangerine peel formula particles, which specifically refers to the method in embodiment 1, and the differences are only that: the preparation methods of the test solution in the step (1) are different.
The method for preparing the test solution of this example includes:
taking a proper amount of dried orange peel formula particles, grinding, accurately weighing 0.2g, placing in a conical flask with a plug, accurately adding 50mL of methanol, weighing, carrying out ultrasonic treatment (power 300W and frequency 40kHz) for 30min, cooling, weighing again, supplementing the lost weight with methanol, shaking uniformly, filtering, and taking a subsequent filtrate to obtain a test solution.
As shown in fig. 2, is a characteristic map of the tangerine peel formula granules (two batches). Wherein 1 is#Peak corresponds to narirutin, 2#The peak is a reference peak corresponding to hesperidin, 5#Peak corresponds to nobiletin, 7#The peak corresponds to hesperetin.
Example 3
The embodiment provides a method for constructing a control characteristic spectrum of a dried orange peel medicinal material, which comprises the following steps:
detecting 10 batches of dried orange peel medicinal materials in different producing areas in table 2 according to the method in the embodiment 1 to obtain 10 characteristic maps; the specific characteristic spectrum of 10 batches of pericarpium Citri Tangerinae is shown in figure 3.
Introducing the 10 obtained characteristic maps into a Chinese medicine chromatogram fingerprint similarity evaluation system, and synthesizing a reference characteristic map, as shown in figure 4. Wherein, the comparison characteristic map has 8 common characteristic peaks, wherein the 1# peak, the 2# peak, the 6# peak and the 8# peak respectively correspond to the peaks of a narirutin control, a hesperidin control, a nobiletin control and an hesperetin control, the 2# peak is taken as a reference peak S, the relative retention time of the 3# peak, the 4# peak, the 5# peak and the 7# peak with the reference peak S is calculated, and the relative peak areas of the 1# peak, the 6# peak and the 8# peak with the reference peak S are respectively as follows:
the relative retention time of the peak 3 is 2.10-2.18; the relative retention time of the 4# peak is 2.95-3.07; the relative retention time of the 5# peak is 3.10-3.22; the relative retention time of the 7# peak is 3.32-3.46;
1#the relative peak area of the peak is not lower than 0.013; the relative peak area of the No. 6 peak is not lower than 0.022; the relative peak area of the 8# peak was not less than 0.007.
TABLE 2 dried orange peel medicinal material sample source information table
| Medicinal material numbering | Producing area |
| S1 | (Hubei) |
| S2 | (Hubei) |
| S3 | Sichuan Jintang |
| S4 | Sichuan Jintang |
| S5 | Yunnan province |
| S6 | Yunnan province |
| S7 | Sichuan charger |
| S8 | Sichuan charger |
| S9 | Four-river eyebrow mountain |
| S10 | Four-river eyebrow mountain |
Example 4
The embodiment provides a method for constructing a control characteristic spectrum of dried orange peel formula particles, which comprises the following steps:
detecting 10 batches of dried orange peel formula granules according to the method in the embodiment 2 to obtain 10 characteristic maps; the specific characteristic spectrum of the dried orange peel granules of 10 batches is shown in figure 5.
Introducing the 10 obtained characteristic maps into a Chinese medicine chromatogram fingerprint similarity evaluation system, and synthesizing a reference characteristic map, as shown in figure 6. Wherein, the comparison characteristic map has 7 common characteristic peaks, the 1# peak, the 2# peak, the 5# peak and the 7# peak respectively correspond to the peaks of a narirutin reference substance, a hesperidin reference substance, a nobiletin reference substance and an hesperetin reference substance, the 2# peak is taken as a reference peak S, the relative retention time of the 3# peak, the 4# peak and the 6# peak with the reference peak S is calculated, and the relative peak areas of the 1# peak, the 5# peak and the 7# peak with the reference peak S are calculated as follows:
3#the relative retention time of the peak is 2.95-3.07; 4#The relative retention time of the peaks is 3.10-3.22; 6#Relative retention of peaksThe interval is 3.32-3.46;
1#the relative peak area of the peak is not lower than 0.06; 5#The relative peak area of the peak is not lower than 0.23; 7#The relative peak area of the peak is not less than 0.05.
Example 5
Methodology investigation of construction method of dried orange peel medicinal material characteristic spectrum
The methodology of the method is verified according to the relevant guiding principle in the current pharmacopoeia, and the construction method of the invention is proved to have good specificity, stability, repeatability and the like.
Comparative example 1
Comparative example 1 reference example 1 is made with the difference that: the mobile phase has different gradient elution, and the obtained characteristic spectrum of pericarpium Citri Tangerinae is shown in FIG. 7.
Specifically, the gradient elution of comparative example 1 was as follows:
| time: min | Mobile phase A: is based on | Mobile phase B: is based on |
| 0～15 | 25 | 75 |
| 15～25 | 25→30 | 75→70 |
| 25～30 | 30→50 | 70→50 |
| 30～35 | 50→60 | 50→40 |
| 35～45 | 60→65 | 40→35 |
| 45～50 | 65→68 | 35→32 |
As can be seen from fig. 7, the mobile phase gradient elution procedure of comparative example 1 gave a non-flat baseline profile and poor resolution.
Finally, it should be noted that: the above embodiments are only used to illustrate the technical solution of the present invention, and not to limit the same; while the invention has been described in detail and with reference to the foregoing embodiments, it will be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; and the modifications or the substitutions do not make the essence of the corresponding technical solutions depart from the scope of the technical solutions of the embodiments of the present invention.
Claims (10)
1. The method for constructing the characteristic map of the dried orange peel medicinal material and the preparation is characterized by comprising the following steps:
detecting the test solution by using an HPLC method to obtain a characteristic spectrum;
the detection conditions of the HPLC method include: taking acetonitrile as a mobile phase A and water as a mobile phase B for gradient elution; the gradient elution comprises: changing the volume fraction of the mobile phase A from 21-23% to 24-26% in 0-15 min; changing the volume fraction of the mobile phase A from 24-26% to 29-31% in 15-25 min; changing the volume fraction of the mobile phase A from 29-31% to 49-51% in 25-30 min; changing the volume fraction of the mobile phase A from 49-51% to 59-61% in 30-35 min; changing the volume fraction of the mobile phase A from 59-61% to 62-63% in 35-45 min; and (4) changing the volume fraction of the mobile phase A from 62-63% to 67-69% in 45-50 min.
2. The method for constructing the characteristic spectrum of the dried orange peel medicinal material and the preparation according to claim 1, which is characterized by further comprising the following steps: detecting the reference substance solution by adopting the HPLC method; the reference substance comprises one or more of narirutin, hesperidin, nobiletin and hesperetin;
preferably, the control comprises narirutin, hesperidin, nobiletin and hesperetin.
3. The method for constructing the characteristic spectrum of the dried orange peel medicinal material and the preparation as claimed in claim 1, wherein in the HPLC method, octadecylsilane chemically bonded silica is used as a filler for a chromatographic column;
and/or in the HPLC method, the detection wavelength is 280-290 nm;
and/or in the HPLC method, the flow rate of the mobile phase is 0.8-1.2 mL/min;
and/or in the HPLC method, the column temperature of a chromatographic column is 25-35 ℃.
4. The method for constructing the characteristic spectrum of the pericarpium citri reticulatae medicinal material and the preparation according to claim 1, wherein the gradient elution is as follows:
5. the method for constructing the characteristic spectrum of the dried orange peel medicinal material and the preparation according to claim 1, wherein the preparation method of the test solution comprises the following steps:
ultrasonically extracting the dried orange peel medicinal material or preparation by using methanol as an extraction solvent, supplementing the loss weight with the extraction solvent, filtering, and taking a subsequent filtrate as a test solution;
preferably, the ultrasonic extraction time is 30-60 min; the power of the ultrasonic wave is 250-350W, and the frequency of the ultrasonic wave is 35-45 kHz.
6. The method for constructing the characteristic spectrum of the pericarpium citri reticulatae medicinal material and the preparation according to claim 5, wherein the material-liquid ratio of the pericarpium citri reticulatae medicinal material to the extraction solvent is (0.1-0.3) g: 25 mL;
and/or the feed-liquid ratio of the preparation to the extraction solvent is (0.1-0.3) g: 50 mL.
7. The method for constructing the characteristic spectrum of the pericarpium citri reticulatae medicinal material and the preparation according to claim 1, wherein the preparation is a pericarpium citri reticulatae single-ingredient preparation;
preferably, the preparation is a dried orange peel formula granule.
8. The method for constructing the characteristic spectrum of the pericarpium citri reticulatae medicinal material and the preparation according to claim 1, wherein the characteristic spectrum of more than 10 batches of pericarpium citri reticulatae medicinal materials or the preparation is constructed according to the HPLC method, and a traditional Chinese medicine chromatogram fingerprint similarity evaluation system is adopted to synthesize the control spectrum of the pericarpium citri reticulatae medicinal material or the preparation.
9. The method for constructing the characteristic spectrum of the pericarpium citri reticulatae medicinal material and the preparation thereof according to claim 8, wherein the control spectrum of the pericarpium citri reticulatae medicinal material has 8 characteristic peaks, wherein the peak 1#, the peak 2#, the peak 6# and the peak 8# respectively correspond to the peaks of a narirutin control, a hesperidin control, a nobiletin control and an hesperetin control, the peak 2# is taken as a reference peak S, the relative retention time of the peak 3#, the peak 4#, the peak 5# and the peak 7# with respect to the reference peak S is calculated, and the relative peak areas of the peak 1#, the peak 6# and the peak 8# with respect to the reference peak S are calculated as follows:
the relative retention time of the peak 3 is 2.10-2.18; the relative retention time of the 4# peak is 2.95-3.07; the relative retention time of the 5# peak is 3.10-3.22; the relative retention time of the 7# peak is 3.32-3.46;
1#the relative peak area of the peak is not lower than 0.013; the relative peak area of the No. 6 peak is not lower than 0.022; the relative peak area of the 8# peak was not less than 0.007.
10. The method for constructing the characteristic spectrum of the pericarpium citri reticulatae medicinal material and the preparation according to claim 8, wherein the reference spectrum of the pericarpium citri reticulatae formula granules has 7 characteristic peaks, wherein the peak 1#, the peak 2#, the peak 5# and the peak 7# respectively correspond to the peaks of a narirutin reference, a hesperidin reference, a nobiletin reference and an hesperetin reference, the peak 2# is taken as a reference peak S, the relative retention time of the peak 3#, the peak 4# and the peak 6# and the reference peak S is calculated, and the relative peak areas of the peak 1#, the peak 5# and the peak 7# and the reference peak S are calculated as follows:
3#the relative retention time of the peak is 2.95-3.07; 4#The relative retention time of the peaks is 3.10-3.22; 6#The relative retention time of the peaks is 3.32-3.46;
1#the relative peak area of the peak is not lower than 0.06; 5#The relative peak area of the peak is not lower than 0.23; 7#The relative peak area of the peak is not less than 0.05.
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| CN101396458A (en) * | 2007-09-28 | 2009-04-01 | 未名天人中药有限公司 | Tangerine peel dispensing granule and preparation method and quality control method thereof |
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| CN101396458A (en) * | 2007-09-28 | 2009-04-01 | 未名天人中药有限公司 | Tangerine peel dispensing granule and preparation method and quality control method thereof |
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