CN113759011A - Method for establishing characteristic spectrum of starwort root and preparation thereof - Google Patents
Method for establishing characteristic spectrum of starwort root and preparation thereof Download PDFInfo
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Abstract
The invention belongs to the technical field of pharmaceutical analysis, and particularly relates to starwort root and a method for establishing a characteristic spectrum of a preparation of the starwort root. The method adopts characteristic chromatogram of radix Stellariae and its preparation established by high performance liquid chromatography, and is suitable for radix Stellariae medicinal material, radix Stellariae decoction pieces, radix Stellariae standard decoction lyophilized powder and radix Stellariae formula granule. The invention can ensure good peak shape and separation degree of characteristic peaks on the characteristic map by controlling the elution gradient program, especially at 9.0-10.5min and 10.5-13.5min, and is helpful for controlling the quality of the dichotoma formula particles. By controlling the elution gradient program, the method can greatly reduce the elution time while ensuring the quality of the characteristic spectrum, improve the analysis speed and save the reagent.
Description
Technical Field
The invention belongs to the technical field of pharmaceutical analysis, and particularly relates to starwort root and a method for establishing a characteristic spectrum of a preparation of the starwort root.
Background
The bupleurum falcatum is dry root of Stellaria dichotoma L.var.lancelata Bge, belonging to Caryophyllaceae, is sweet in taste and slightly cold, enters liver and stomach channels, has the effects of clearing deficiency heat and removing infantile malnutrition heat, is used for treating symptoms such as fever due to yin deficiency, tuberculosis heat due to bone steaming, infantile malnutrition heat and the like, and is one of main medicaments of famous Chinese patent medicines such as black-bone chicken white phoenix pills and the like.
At present, the application forms of Stellaria dichotoma are mainly Stellaria dichotoma medicinal materials, Stellaria dichotoma decoction pieces, Stellaria dichotoma standard decoction freeze-dried powder, Stellaria dichotoma formula granules and a part of compound medicines containing Stellaria dichotoma. The starwort root standard decoction freeze-dried powder is a freeze-dried powder obtained by carrying out freeze drying on starwort root decoction pieces after vacuum concentration according to the decoction requirement of the traditional decoction, and the components of the freeze-dried powder are consistent with those of the standard decoction; the traditional Chinese medicine formula particle is a novel medicine which is prepared by taking traditional Chinese medicine decoction pieces as raw materials through production processes of extraction, concentration, drying, granulation and the like and is prepared into a certain specification and used for clinical formula of medical institutions. It is based on the standard decoction of traditional Chinese medicine and needs to be used under the guidance of the theory of traditional Chinese medicine. Compared with traditional Chinese medicine decoction, the decoction has the advantages of no decoction, easy administration, easy storage, convenient carrying, controllable quality, etc.
2015 edition of Chinese pharmacopoeia records the quality standard of Stellaria delavayi Diels and decoction pieces, but has no index component and characteristic spectrum detection item. There is no standard Bupleurum scorzonerifolium reference medicinal material on the market, which increases the difficulty of controlling the quality of Bupleurum scorzonerifolium. The bupleurum root formula granules are lack of uniform quality standards, and the research reports are less.
The prior art discloses two identification methods of ultraviolet spectrophotometry and thin-layer chromatography of the starwort root formula granules, but the prior art is not enough to comprehensively control the quality of the starwort root formula granules. The characteristic spectrum is one of effective methods widely used for evaluating the quality of the traditional Chinese medicine in recent years, and the authenticity and the consistency and the stability of the quality of the traditional Chinese medicine and the preparation thereof can be effectively controlled by adopting a comprehensive and quantifiable identification mode. The preliminary research on the characteristic spectrum of the starwort root high performance liquid chromatography disclosed by Liu Jiang hong et al discloses that the characteristic spectrum of the starwort root is established by adopting the high performance liquid chromatography, but the preparation steps of the sample solution in the research are complicated, the Soxhlet extraction time by adding methanol is 24h, the extraction is carried out for 10 times by using ether, the extraction time is longer, toxic reagents are adopted, the polarity of the extracted components is smaller, and the preparation method and the chromatographic detection conditions of the sample solution in the research are not suitable for detecting the starwort root preparation extracted by water decoction.
Disclosure of Invention
Therefore, the invention aims to overcome the defects that the prior art does not have a method for establishing a liquid phase characteristic spectrum of a starwort root water extract preparation such as a starwort root formula particle and the like, and provides a method for establishing a characteristic spectrum of the starwort root and the preparation thereof.
Therefore, the invention provides the following technical scheme.
The invention provides a method for establishing a characteristic spectrum of Stellaria dichotoma and a preparation thereof, which comprises the steps of obtaining the characteristic spectrum of Stellaria dichotoma and the preparation thereof, wherein the characteristic spectrum comprises 4 common characteristic peaks which are respectively as follows: peak No. 1, peak No. 2, peak No. 3, peak No. 4; taking No. 1 common characteristic peak dichotomamine B as a reference peak, wherein the relative retention time of the rest characteristic peaks is within +/-10% of a specified value, and the specified values of the relative retention time of the characteristic peaks are respectively as follows:
peak No. 4 had a relative retention time of 2.02.
The establishing method comprises the following steps of,
preparation of a test solution: taking a sample to be detected of starwort root to prepare a test solution;
chromatographic conditions are as follows: performing high performance liquid chromatography with octadecylsilane chemically bonded silica as filler; using acetonitrile as a mobile phase A and acetic acid as a mobile phase B, and performing gradient elution, wherein the gradient elution procedure comprises the following steps: 0-1.5min, mobile phase A: the mobile phase B is 0.7-1.5%: 99.3 to 98.5 percent; 1.5-3.5min, mobile phase A: 1.5-5.0% of mobile phase B: 98.5-95.0%; 3.5-4.0min, mobile phase A: 5.0-10.0% of mobile phase B: 95.0 to 90.0 percent; 4.0-9.0min, mobile phase A: 10.0-13.5% of mobile phase B: 90.0-86.5%; 9.0-10.5min, mobile phase A: 13.5-12.0% of mobile phase B: 86.5 to 88.0 percent; 10.5-13.5min, mobile phase A: the mobile phase B is 12.0-26.0%: 88.0 to 74.0 percent; 13.5-18.5min, mobile phase A: 26.0-43.0% of mobile phase B: 74.0-57.0%; the detection wavelength is 250-290 nm; the flow rate is 0.2-0.7 ml/min;
and (3) determination: and sucking the test solution, injecting into an ultra-high performance liquid chromatograph, and measuring to obtain the test solution.
The chromatographic conditions are as follows: performing high performance liquid chromatography with octadecylsilane chemically bonded silica as filler and detection wavelength of 260-280 nm; the flow rate is 0.3-0.5 ml/min; the column temperature is 29-31 ℃; using acetonitrile as mobile phase A and using volume fraction of 0.01-0.1% acetic acid as mobile phase B, and carrying out gradient elution.
The preparation of the test solution comprises the following specific steps,
grinding radix Stellariae sample to be tested, precisely weighing, extracting with water for 10-30min, filtering, and collecting the filtrate.
The concentration of the test solution is 40-75 mg/ml.
The extraction method is an ultrasonic extraction method or a reflux extraction method.
The Stellaria dichotoma sample to be detected is Stellaria dichotoma medicinal material, Stellaria dichotoma decoction pieces, Stellaria dichotoma standard decoction freeze-dried powder or Stellaria dichotoma formula granules.
The technical scheme of the invention has the following advantages:
1. the method adopts the characteristic spectrum of the starwort root and the preparation thereof established by the high performance liquid chromatography, and is suitable for establishing the characteristic spectrum of the starwort root water extract preparation such as formula granules; the characteristic spectrum obtained by using the starwort root amine B as a reference peak can carry out multi-component and integral quality control on starwort root medicinal materials, starwort root decoction pieces, starwort root standard decoction freeze-dried powder and starwort root formula granules, and the method is verified by methodological experiments to have high accuracy and good repeatability.
The invention can ensure good peak shape and separation degree of characteristic peaks on a characteristic map by controlling an elution gradient program, especially at 9.0-10.5min and 10.5-13.5min, and is beneficial to controlling the quality of Stellaria dichotoma and preparations thereof. By controlling the elution gradient program, the method can greatly reduce the elution time while ensuring the quality of the characteristic spectrum, improve the analysis speed and save the reagent.
The method is established based on standard decoction material, ensures consistency of radix Stellariae aqueous extract preparation and standard decoction, and has objective and accurate quality control.
2. According to the method for establishing the characteristic spectrum of the dichotoma and the preparation thereof, the preparation method of the test solution is simple and rapid, water is used as a solvent, the use of toxic reagents is avoided, and the safety and the environmental friendliness in the characteristic spectrum establishing process are improved. The method has the advantages that the overall analysis time is controlled within 1h, the analysis time is greatly shortened, the working efficiency is improved, and the labor and material cost is saved.
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 18 batches of Standard decoction lyophilized powder of Stellaria dichotoma in example 1 of the invention;
FIG. 2 is a comparison feature map generated from 18 batches of Standard decoction of Bupleurum scorzonerifolium freeze-dried powder in example 1 of the present invention;
FIG. 3 is a 3D full scan of a test solution in Experimental example 5 of the present invention;
FIG. 4 is a characteristic map of a test solution of test group 1 in Experimental example 6 of the present invention;
FIG. 5 is a characteristic map of a test solution of the control group 1 in Experimental example 6 of the present invention;
FIG. 6 is a characteristic map of a test solution of test group 1 in Experimental example 7 of the present invention;
FIG. 7 is a characteristic map of a test solution of control group 1 in Experimental example 7 of the present invention;
FIG. 8 is a characteristic map of a test solution of control group 2 in Experimental example 7 of the present invention;
FIG. 9 is a characteristic map of a test solution of the control group 1 in Experimental example 8 of the present invention;
FIG. 10 is a characteristic map of a test solution of control group 2 in Experimental example 8 of the present invention;
FIG. 11 is a characteristic map of a test solution in Experimental example 9 of the present invention;
FIG. 12 is a characteristic diagram of a Scorzonera dichotoma sample to be tested in Experimental example 10 of the present invention.
Detailed Description
The following examples are provided to further understand the present invention, not to limit the scope of the present invention, but to provide the best mode, not to limit the content and the protection scope of the present invention, and any product similar or similar to the present invention, which is obtained by combining the present invention with other prior art features, 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 not indicated by manufacturers, and are all conventional reagent products which can be obtained commercially.
The invention relates to a reagent and an instrument:
reagent testing:
the preparation method of the starwort standard decoction freeze-dried powder comprises the following steps: soaking radix Stellariae decoction pieces in casserole for 30min, decocting twice, adding water 9 times the amount of decoction pieces in one decoction, boiling with strong fire, decocting with slow fire for 20min, filtering while hot, and rapidly cooling; adding water 7 times the amount of decoction pieces, boiling with strong fire, decocting with slow fire for 15min, filtering, mixing filtrates, concentrating, and freeze drying to obtain the final product; the invention uses 18 batches of starwort root standard decoction freeze-dried powder which are respectively marked as S1-S1, the 18 batches of freeze-dried powder are only different in producing areas of starwort root decoction pieces, and the producing areas of the decoction pieces corresponding to the 18 batches of starwort root standard decoction freeze-dried powder are as follows:
170122 origin 748411-01: gansu province regaining county ten miles town;
170122 origin 748103-02: shansu Longxi county, Ma river Town;
170122 origin of 748106-03: gansu province Longxi county double-spring village gift sheep mouth village;
170327 seed 743000-04 origin: dingxi city, gansu province;
170327 origin of 748100-05: longxi county, Dengxi city, Gansu province;
170327 and 756000-06 production places: ningxia Hui nationality Guyuan City;
170327 seed 741000-07 production area: tianshui city of Gansu province;
170406 + 756300-08 origin: the Shen Lin Xiang of Longde county, Guyuan city, Hui nationality of Ningxia;
170406 + 756500-09 origin: city, yang, county, city, county, time, village, etc., in the city of origin, guoyang, national province of the Ningxia Hui nationality;
170406 and 756500-10 production places: city, yang, county, etc. of the city of the national treasures of the national province of the Ningxia Hui;
170421 + 751300-11 production areas: wuzhou concentric county in Ningxia Hui nationality autonomous region;
170421 and 751300-12 production areas: wuzhou concentric county in Ningxia Hui nationality autonomous region;
170421 and 751300-13 production areas: wuzhou concentric county in Ningxia Hui nationality autonomous region;
170525 onset 753204-14 origin: xixia Shizu mountain city Wen Yongguxiang;
170525 and 753400-15 production places: ningxia Pingluo county, Liuzhongxiang;
170525 region of origin 750200-16: ningxia Helan county Tongxiang;
170525 region of origin 750200-17: ningxia Helan county Jingui Zhen;
170525 and 753400-18 production places: ningxia Pinluo county at the canal opening village.
The preparation method of the starwort root formula particle comprises the following steps: decocting radix Stellariae decoction pieces (produced area is Min county and Ten Li town of Gansu province) with 12 times of water for 1.5h, and filtering to obtain filtrate; decocting the two decoctions in 10 times of water for 1 hr, filtering, mixing filtrates, concentrating the filtrate under reduced pressure to obtain extract, drying, adding dextrin, granulating, and packaging. The decoction pieces for preparing the starwort root formula particle of the invention have the batch number of 170122-.
Reagent:
formic acid (national pharmaceutical group chemical reagents ltd), chromatographically pure;
phosphoric acid (national drug group chemical reagents limited), chromatographically pure;
acetonitrile (merck), chromatographically pure;
methanol (national chemical group chemical reagent, ltd), analytically pure;
ethanol (national chemical group chemical reagent, ltd), analytically pure;
acetic acid (national chemical group, chemical Co., Ltd.), analytically pure;
the water is distilled water (drochen).
The instrument comprises the following steps:
ACQUITY
an H-Class ultra-high performance liquid chromatograph, a PDA Detector, a TUV Detector and an Empower 3 chromatographic workstation;
ME104E electronic balance (mettler-toledo);
KQ-300DB ultrasonic cleaner (Kunshan ultrasonic instruments Co., Ltd.).
Example 1
The embodiment provides a method for establishing a characteristic spectrum of Stellaria dichotoma and a preparation thereof, which comprises the following steps,
preparation of a test solution: taking a sample to be tested of Stellaria dichotoma, grinding, taking 1.0g, precisely weighing, placing in a conical flask with a plug, precisely adding 20ml of water, sealing the plug, carrying out ultrasonic treatment for 20min, wherein the ultrasonic power is 250W, the frequency is 40kHz, taking out, cooling, shaking up uniformly, filtering, and taking a subsequent filtrate to obtain the final product, wherein the concentration of the sample solution is 50 mg/ml.
Chromatographic conditions are as follows: performing high performance liquid chromatography with octadecylsilane chemically bonded silica as filler, and ACQUITY UPLC HSS T3 (column length of 7.5cm, column inner diameter of 2.1mm, and particle diameter of 1.8 μm) as chromatographic column; using acetonitrile as a mobile phase A and acetic acid with the volume fraction of 0.05% as a mobile phase B, and performing gradient elution, wherein the gradient elution procedure comprises the following steps: 0-1.5min, mobile phase A: the mobile phase B is 0.7-1.5%: 99.3 to 98.5 percent; 1.5-3.5min, mobile phase A: 1.5-5.0% of mobile phase B: 98.5-95%; 3.5-4.0min, mobile phase A: 5.0-10.0% of mobile phase B: 95.0 to 90.0 percent; 4.0-9.0min, mobile phase A: 10.0-13.5% of mobile phase B: 90.0-86.5%; 9.0-10.5min, mobile phase A: 13.5-12.0% of mobile phase B: 86.5 to 88.0 percent; 10.5-13.5min, mobile phase A: the mobile phase B is 12.0-26.0%: 88.0 to 74.0 percent; 13.5-18.5min, mobile phase A: 26.0-43.0% of mobile phase B: 74.0-57.0%; the detection wavelength is 270 nm; the flow rate is 0.4 ml/min; the column temperature was 30 ℃.
And (3) determination: sucking the sample solution, injecting into ultra high performance liquid chromatograph with injection amount of 2 μ L, and measuring.
Identification of characteristic peaks
Respectively taking 18 batches of starwort root standard decoction freeze-dried powder, preparing the test solution of the 18 batches of starwort root standard decoction freeze-dried powder according to the item of 'preparation of the test solution', and then obtaining the characteristic map of each batch of test solution according to the item of 'chromatographic conditions' and 'determination', wherein the figure is 1; selecting 4 peaks with better repeatability as common peaks, namely a common characteristic peak No. 1, a common characteristic peak No. 2, a common characteristic peak No. 3 and a common characteristic peak No. 4, calculating the relative retention time and the relative peak area of other characteristic peaks by taking the common characteristic peak No. 1 as a reference peak (S peak), and referring to tables 1 and 2. As can be seen from tables 1 and 2, the relative retention time differences of the characteristic peaks are small and are within the range of +/-10%, the quality control requirements are met, and the average value of the relative retention time is selected as a measured value; the measured values were: 1 (peak 1S), 1.24 (peak 2), 1.28 (peak 3), 2.02 (peak 4), error allowance: 10% of the total weight of the composition. The difference of the relative peak areas is large, the RSD value is 8.8-26.0%, and the quality control requirement is not met, so that the relative peak areas are not specified.
The characteristic map of the starwort root and the preparation thereof comprises 4 common characteristic peaks, wherein the No. 1 peak is taken as a reference peak (S peak), and the relative retention time of other characteristic peaks is within +/-10% of a specified value, which are respectively as follows:
peak 2 had a relative retention time of 1.24;
peak 3 had a relative retention time of 1.28;
peak No. 4 had a relative retention time of 2.02.
TABLE 118 batches of Standard decoction of Stellaria dichotoma freeze-dried powder characteristic peak retention time and relative retention time
TABLE 218 peak area and relative peak area of characteristic peak of Stellaria dichotoma standard decoction lyophilized powder
Using a traditional Chinese medicine chromatogram fingerprint similarity evaluation system (version 2012.1) to generate a starwort root standard decoction freeze-dried powder comparison characteristic spectrum, wherein the comparison characteristic spectrum is shown in figure 2, the similarity result is shown in table 3, and R in figure 1 is a comparison characteristic spectrum, so that comparison with the spectrum of 18 batches of starwort root standard decoction freeze-dried powder is facilitated.
TABLE 3 results of similarity calculation
| Numbering | Batch number | Similarity to control |
| 1 | 170122-748411-01 | 1.000 |
| 2 | 170122-748103-02 | 1.000 |
| 3 | 170122-748106-03 | 1.000 |
| 4 | 170327-743000-04 | 0.996 |
| 5 | 170327-748100-05 | 0.989 |
| 6 | 170327-756000-06 | 0.998 |
| 7 | 170327-741000-07 | 1.000 |
| 8 | 170406-756300-08 | 0.966 |
| 9 | 170406-756500-09 | 0.961 |
| 10 | 170406-756500-10 | 1.000 |
| 11 | 170421-751300-11 | 1.000 |
| 12 | 170421-751300-12 | 0.998 |
| 13 | 170421-751300-13 | 1.000 |
| 14 | 170525-753204-14 | 1.000 |
| 15 | 170525-753400-15 | 0.984 |
| 16 | 170525-750200-16 | 1.000 |
| 17 | 170525-750200-17 | 0.991 |
| 18 | 170525-753400-18 | 0.989 |
| - | R | 1.000 |
As can be seen in Table 3, the similarity between the characteristic spectrum of 18 batches of starwort root standard decoction freeze-dried powder and the comparison characteristic spectrum is 0.961-1 and is higher than 0.95, which indicates that the difference of the characteristic spectrums of each batch of starwort root standard decoction freeze-dried powder is small.
Identification of characteristic peaks
Taking starwort root standard decoction freeze-dried powder (batch number is S1), preparing the test solution according to the item of the preparation of the test solution, then obtaining a mass spectrogram of the test solution by using a high performance liquid chromatography-tandem high resolution time-of-flight mass spectrometer according to the chromatographic condition, and carrying out structural determination on 4 characteristic peaks, wherein the molecular formula of each characteristic peak is shown in Table 4.
Table 4 results of mass spectrum component estimation
Experimental example 1 examination of extraction solvent in preparation of test sample solution
Taking the same batch of starwort root standard decoction freeze-dried powder (batch number is S1), and taking 3 parts in total.
The preparation method of the test solution in the experimental group 1 comprises taking starwort root standard decoction lyophilized powder, grinding, taking 1.0g, precisely weighing, placing in a conical flask with a plug, precisely adding 20ml of water, sealing the plug, performing ultrasonic treatment for 20min with ultrasonic power of 250W and frequency of 40kHz, taking out, cooling, shaking up, filtering, and taking out the subsequent filtrate; then, the characteristic spectrum of the test solution was obtained under the terms "chromatographic conditions" and "measurement" in example 1.
The control group 1 was different from the experimental group 1 only in that methanol was used as an extraction solvent instead of water in the experimental group 1.
The control group 2 was different from the experimental group 1 only in that ethanol was used as an extraction solvent instead of water in the experimental group 1.
The peak areas of the characteristic peaks in the characteristic patterns obtained from the experimental group 1, the control group 1 and the control group 2 are shown in table 5.
TABLE 5 Peak areas of characteristic peaks of characteristic spectra of experimental group and control group
| Examples of the invention | Peak to peak area of No. 1 | Peak to peak area of No. 2 | Peak to peak area of No. 3 | Peak to peak area of No. 4 |
| |
1523707 | 123707 | 106908 | 214198 |
| |
457239 | 36286 | 9464 | 48767 |
| |
2878412 | 212192 | 199248 | 346088 |
| RSD(％) | 74.9 | 70.9 | 90.2 | 73.4 |
Table 5, substance information represented by the characteristic patterns obtained by different extraction solvents is substantially the same, and it can be known from RSD of peak areas of the characteristic peaks that the characteristic peaks are greatly influenced by the solvents, where the peak areas of the characteristic peaks obtained by using water as a solvent are higher and water is non-toxic, and water is selected as an extraction solvent for preparing a sample solution according to the principle that the sample preparation toxicity is the lowest.
Experimental example 2 examination of extraction method in preparation of test solution
Taking the same batch of starwort standard decoction freeze-dried powder (batch number is S1), and taking 2 parts in total.
The preparation method of the test solution in the experimental group 1 comprises taking starwort root standard decoction lyophilized powder, grinding, taking 1.0g, precisely weighing, placing in a conical flask with a plug, precisely adding 20ml of water, sealing the plug, performing ultrasonic treatment for 30min with ultrasonic power of 250W and frequency of 40kHz, taking out, cooling, shaking up, filtering, and taking out the subsequent filtrate; then, the characteristic spectrum of the test solution was obtained under the terms "chromatographic conditions" and "measurement" in example 1.
The preparation method of the test solution in control group 1 comprises collecting starwort root standard decoction lyophilized powder, grinding, collecting 1.0g, precisely weighing, placing in a conical flask with a plug, precisely adding 20ml of water, sealing the plug, heating under reflux for 30min, taking out, cooling, shaking, filtering, and collecting the filtrate; then, the characteristic spectrum of the test solution was obtained under the terms "chromatographic conditions" and "measurement" in example 1.
The peak areas of the characteristic patterns of the experimental group 1 and the control group 1 are shown in Table 6.
TABLE 6 Peak areas of characteristic peaks of characteristic spectra of experiment group 1 and control group 1
| Examples of the invention | Peak to peak area of No. 1 | Peak to peak area of No. 2 | Peak to peak area of No. 3 | Peak to peak area of No. 4 |
| |
2841589 | 187760 | 187708 | 349746 |
| |
2891265 | 193657 | 200348 | 346474 |
| RSD(％) | 1.2 | 2.2 | 4.6 | 0.7 |
Table 6 shows that the peak areas RSD of the peak 3 in the feature maps obtained by the two extraction methods are greatly different, the peak area of the peak 3 of the feature map obtained by ultrasonic extraction is higher than that of the reflow method, the peak areas RSD of the other feature peaks are slightly different, and the ultrasonic is selected as the extraction method in consideration of the convenience of experimental operation.
Experimental example 3 examination of concentration of sample solution
Taking 6 parts of starwort root standard decoction freeze-dried powder (batch number is S1) of the same batch. Experimental group 1 a sample solution was prepared according to the method under the item "preparation of sample solution" in example 1, and then a characteristic map was created according to the conditions of the item "chromatographic conditions" and the method of the item "determination" in example 1.
The control group 1 is different from the experimental group 1 only in the concentration of the test solution, and the concentration of the test solution of the control group 1 is 10 mg/ml.
The control group 2 is different from the experimental group 1 only in the concentration of the test solution, and the concentration of the test solution of the control group 2 is 25 mg/ml.
The control group 3 is different from the experimental group 1 only in the concentration of the test solution, and the concentration of the test solution of the control group 3 is 35 mg/ml.
The control group 4 was different from the experimental group only in the concentration of the test solution, and the concentration of the test solution of the control group 4 was 75 mg/ml.
The control group 5 is different from the experimental group only in the concentration of the test solution, and the concentration of the test solution of the control group 5 is 100 mg/ml.
The peak areas of the characteristic patterns obtained from the experimental group 1 and the control groups 1 to 5 are shown in table 7, and the peak area of the No. 1 peak in the characteristic peaks is higher, so that the heights of the No. 1 peaks of the sample solutions with different concentrations are compared, and the result is shown in table 8.
TABLE 7 examination of the concentration of the test solution
TABLE 8 comparison of peak heights in feature Profile No. 1
According to the principle that the peak height range of the ultra-high performance liquid chromatogram is 0.5-1.0AU, the optimal height is 0.5AU, so that the optimal concentration is 50mg/ml of the solution of the test sample.
Experimental example 4 examination of extraction time
Taking 3 parts of starwort root standard decoction freeze-dried powder (batch number is S1) of the same batch, preparing the test solution by the experimental group 1 according to the method under the item of 'preparation of test solution' in the embodiment, and then establishing a characteristic map according to the condition of the item of 'chromatographic condition' and the method of the item of 'determination' in the embodiment 1.
The control group 1 is different from the experimental group 1 only in the ultrasonic extraction time, and the ultrasonic extraction time is 30min when the test solution is prepared in the control group 1.
The control group 2 is different from the experimental group 1 only in the ultrasonic extraction time, and the ultrasonic extraction time of the control group 2 is 50min when the test solution is prepared.
The peak areas of the characteristic patterns of the experimental group 1, the control group 1 and the control group 2 are shown in Table 9.
TABLE 9 extraction of time profiles
The results in Table 9 show that the extraction time has a small influence on the peak area, and the extraction time of the present invention is determined to be 20min in order to save time.
Experimental example 5 examination of detection wavelength
Taking starwort root standard decoction freeze-dried powder (batch number S1), preparing the test solution according to the item of 'preparation of the test solution' in the embodiment 1, and then carrying out 3D full scanning on the test solution, wherein the result shown in figure 3 shows that the peak around 270nm is rich, the peak shape is better and is better than other wavelengths, so 270nm is selected as the detection wavelength.
Experimental example 6 examination of column
Taking the same batch of starwort root standard decoction freeze-dried powder (batch number is S1), and preparing the test solution according to the item of the preparation of the test solution in the embodiment 1 in total 2 parts.
Chromatographic conditions for experimental group 1: performing high performance liquid chromatography with octadecylsilane chemically bonded silica as filler, and ACQUITY UPLC HSS T3 (column length of 7.5cm, column inner diameter of 2.1mm, and particle diameter of 1.8 μm) as chromatographic column; using acetonitrile as a mobile phase A and acetic acid with the volume fraction of 0.05% as a mobile phase B, and performing gradient elution, wherein the gradient elution procedure comprises the following steps: 0-4min, mobile phase A: the mobile phase B is 2%: 98 percent; 4-19min, mobile phase A: 2-19% of mobile phase B: 98-81%; 19-21min, mobile phase A: the mobile phase B is 19-30%: 81-70 percent; 21-25min, mobile phase A: 30-46% of mobile phase B: 70-54%; 25-30min, mobile phase A: 46-75% of mobile phase B: 54 to 25 percent; the detection wavelength is 270 nm; the flow rate is 0.4 ml/min; the column temperature is 30 ℃, then the sample solution is absorbed and injected into the ultra-high performance color phase chromatograph, the injection amount is 2 mu L, and the characteristic spectrum is obtained by determination.
The characteristic maps of the experimental group 1 and the control group 1 are respectively shown in fig. 4 and fig. 5, and the results show that the acquired UPLC HSS T3 chromatographic column can tolerate 100% of water phase and is favorable for gradient adjustment of elution gradient, so the chromatographic column is selected.
Experimental example 7 examination of Mobile phase
Taking the same batch of starwort root standard decoction freeze-dried powder (batch number is S1), preparing the test solution according to the item of the preparation of the test solution in the embodiment 1, and obtaining 3 parts in total.
Experimental group 1A characteristic map of a test solution was obtained under the terms "chromatographic conditions" and "assay" in example 1.
The control 1 differs from the experimental group 1 only in that the control 1 has a volume fraction of 0.05% formic acid as the mobile phase B.
The control group 2 was different from the experimental group 1 only in that the control group 2 had a volume fraction of 0.05% phosphoric acid as the mobile phase B.
The characteristic maps of the experimental group 1, the control group 1 and the control group 2 are shown in fig. 6-8, and the results show that the peak shapes of the chromatographic peaks of the obtained characteristic maps are most symmetrical and the separation degree is the best and is obviously better than that of 0.05% formic acid and 0.05% phosphoric acid when 0.05% acetic acid is used as the mobile phase B, so that 0.05% acetic acid is selected as the mobile phase B.
Experimental example 8 examination of elution gradient
Taking 3 parts of starwort root standard decoction freeze-dried powder (batch number is S1) of the same batch, and preparing the test solution according to the item of the preparation of the test solution in the embodiment 1.
Experimental group 1A characteristic map of a test solution was obtained under the terms "chromatographic conditions" and "assay" in example 1.
Chromatographic conditions for control 1: performing high performance liquid chromatography with octadecylsilane chemically bonded silica as filler, and ACQUITY UPLCHSS T3 (column length of 7.5cm, column inner diameter of 2.1mm, and particle diameter of 1.8 μm) as chromatographic column; using acetonitrile as a mobile phase A and acetic acid with volume fraction of 0.01-0.1% as a mobile phase B, and performing gradient elution, wherein the gradient elution procedure comprises the following steps: 0-4min, mobile phase A: mobile phase B is 0%: 100 percent; 4-22min, mobile phase A: the mobile phase B is 0-15%: 100 to 85 percent; 22-24min, mobile phase A: 15-30% of mobile phase B: 85 to 70 percent; 24-28min, mobile phase A: 30-43% of mobile phase B: 70-57%; 28-35min, mobile phase A: the mobile phase B is 43-75%: 57-25%; the detection wavelength is 270 nm; the flow rate is 0.4 ml/min; the column temperature is 30 ℃; then obtaining the characteristic map of the test solution according to the item of 'determination'.
Chromatographic conditions for control 2: performing high performance liquid chromatography with octadecylsilane chemically bonded silica as filler; using acetonitrile as a mobile phase A and acetic acid as a mobile phase B, and performing gradient elution, wherein the gradient elution procedure comprises the following steps: 0-4min, mobile phase A: mobile phase B is 0%: 100 percent; 4-6.5min, mobile phase A: the mobile phase B is 0-1.5%: 100 to 98.5 percent; 6.5-7.0min, mobile phase A: 1.5-4.0% of mobile phase B: 98.5 to 96 percent; 7.0-8.0min, mobile phase A: the mobile phase B is 4.0-5.0%: 96.0 to 95.0 percent; 8.0-8.5min, mobile phase A: 5.0-10.0% of mobile phase B: 95.0 to 90.0 percent; 8.5-14.0min, mobile phase A: 10.0-14.0% of mobile phase B: 90.0-86.0%; 14.0-16.0min, mobile phase A: the mobile phase B is 14.0-30.0%: 86.0 to 70.0 percent; 16.0-20.0min, mobile phase A: 30.0-43.0% of mobile phase B: 70.0 to 57.0 percent; the detection wavelength is 270 nm; the flow rate is 0.4 ml/min; the column temperature is 30 ℃; then obtaining the characteristic map of the test solution according to the item of 'determination'.
The characteristic map of the experimental group 1 is as shown in FIG. 6, the characteristic map of the control group 1 is shown in FIG. 9, and the characteristic map of the control group 2 is shown in FIG. 10.
The results show that the peak shape separation degree of the characteristic spectrum obtained by the elution gradient in the control group 1 is poor between 20 and 28min, and the fluctuation of the base line is large. The control group 2 has fewer peaks in 0-9min, the peak shape separation degree in 14.5-19min is poor, and the characteristic spectrum obtained by the experimental group 1 has better peak shape and separation degree, so the gradient elution program in the experimental group 1 is selected.
Experimental example 9 examination of delay Property
Taking radix bupleuri standard decoction lyophilized powder (batch number is S1) as a test solution, preparing the test solution according to the item of 'preparation of test solution' in example 1, then injecting sample according to the 'chromatographic condition' in example 1, and recording the characteristic spectrum of 2 times of gradient elution time, as shown in figure 11.
The results show that: after 18.5min no significant lag peaks appeared and the chromatographic method met the analytical requirements.
Experimental example 10 examination of Bupleurum scorzonerifolium Linn samples to be tested
Experimental group 1: taking a sample to be tested of the starwort root medicinal material, grinding, taking 1.0g, precisely weighing, placing in a conical flask with a plug, precisely adding 20ml of water, sealing the plug, carrying out ultrasonic treatment for 20min, wherein the ultrasonic power is 250W, the frequency is 40kHz, taking out, cooling, shaking up, filtering, and taking a subsequent filtrate to obtain the sample solution with the concentration of 50 mg/ml.
Experimental group 2: taking a sample to be tested of the starwort decoction pieces, grinding, taking 1.0g, precisely weighing, placing in a conical flask with a plug, precisely adding 20ml of water, sealing the plug, carrying out ultrasonic treatment for 20min, wherein the ultrasonic power is 250W, the frequency is 40kHz, taking out, cooling, shaking up, filtering, and taking a subsequent filtrate to obtain the sample solution with the concentration of 50 mg/ml.
Experimental group 3: taking a sample to be tested of the starwort root formula particles, grinding, taking 1.0g, precisely weighing, placing in a conical flask with a plug, precisely adding 20ml of water, sealing the plug, carrying out ultrasonic treatment for 20min, wherein the ultrasonic power is 250W, the frequency is 40kHz, taking out, cooling, shaking up, filtering, and taking a subsequent filtrate to obtain the sample solution with the concentration of 50 mg/ml.
The sample solutions prepared in the experimental groups 1 to 3 were injected according to the "chromatographic conditions" of example 1, the injection amount was 2. mu.L, and a characteristic spectrum, as shown in FIG. 12, was obtained by measurement, and the relative retention times were shown in Table 10.
TABLE 10 relative retention time tables of Bupleurum scorzonerifolium herbs, decoction pieces, and granules
| Name (R) | Peak 1(S peak) | |
|
|
| Bupleurum scorzonerifolium medicinal material 170122- | 1 | 1.245 | 1.278 | 2.020 |
| Bupleurum scorzonerifolium decoction pieces 170122- | 1 | 1.245 | 1.276 | 2.020 |
| Radix Stellariae granules170122-748411-01 | 1 | 1.245 | 1.276 | 2.022 |
| Standard of |
1 | 1.24 | 1.28 | 2.02 |
As can be seen from fig. 12 and table 10, the relative retention time of the starwort root medicinal material, the starwort root decoction pieces and the starwort root formula granules is within ± 10% of the specified value, which meets the requirement, and the establishment method of the characteristic spectrum of the starwort root and the preparation thereof provided by the invention is suitable for the starwort root medicinal material, the starwort root decoction pieces, the starwort root standard decoction freeze-dried powder and the starwort root formula granules.
Example 2
This example provides a methodological review of the method for establishing the characteristic spectrum of Stellaria dichotoma and its preparation in example 1.
(1) Precision survey
Repeatability survey
Taking 6 parts of starwort root standard decoction freeze-dried powder (batch number: S1) of the same batch, preparing the test solution according to the item of 'preparation of the test solution' in the example 1, then obtaining a characteristic map of the test solution according to the item of 'chromatographic conditions' and 'determination' in the example 1, calculating the relative peak area and the relative retention time of the peak with the peak number 1 as a reference peak, and calculating RSD, wherein the results are shown in tables 11 and 12.
TABLE 11 relative retention time of characteristic peaks
TABLE 12 relative peak area of characteristic peaks
From the results of tables 11 and 12, the relative retention time RSD of each characteristic peak was in the range of 0.1% to 0.2%, and the relative peak area RSD was in the range of 0.7% to 1.9%, indicating that the reproducibility of the characteristic pattern was good.
Intermediate precision investigation
Taking 6 parts of starwort root standard decoction freeze-dried powder (batch number is S1) of the same batch, preparing a sample solution according to the item of 'preparation of the sample solution' in the example 1, then obtaining a characteristic map of the sample solution under a Waters UPLC H-Class and TUV detector according to the 'chromatographic condition' in the example 1, calculating the relative peak area and the relative retention time of the sample solution by taking the peak 1 as a reference peak, and calculating RSD, wherein the result is shown in a table 13 and a table 14.
TABLE 13 relative retention time of characteristic peaks
TABLE 14 relative peak area of characteristic peaks
In the characteristic spectrum obtained by using a Waters UPLC H-Class TUV detector, the relative retention time RSD of 4 characteristic peaks is in the range of 0.1-0.2%, and the relative peak area RSD is in the range of 0.8-3.4%. The RSD range of the relative retention time between different instruments is 0.6-2.4%, and the relative peak area range is 3.2-4.2%, which indicates that only the relative retention time is investigated between different instruments, and the relative retention time meets the analysis requirements.
(2) Stability survey
Taking 8 parts of starwort root standard decoction freeze-dried powder (batch number is S1) of the same batch, preparing the test solution according to the item of 'preparation of the test solution' in the embodiment 1, respectively placing the test solution for 0, 2, 4, 6, 8, 10, 12 and 24 hours at room temperature, then obtaining the characteristic spectrum of the test solution according to the item of 'chromatographic condition' and 'determination' in the embodiment 1, taking the peak No. 1 as a reference peak, calculating the relative peak area and the relative retention time, and calculating RSD, wherein the results are shown in a table 15 and a table 16.
TABLE 15 relative retention time of characteristic peaks
TABLE 16 relative peak area of characteristic peaks
As can be seen from the results of the stability tests in tables 15 and 16, the RSD of the 4 characteristic peaks is in the range of 0.2% to 0.5% and the RSD of the relative peak area is in the range of 0.3% to 1.6% when the solution stability is examined for 24 hours. The characteristic spectrum analysis method adopted by the invention is stable, reliable and good in reproducibility.
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 (7)
1. A method for establishing a characteristic spectrum of Stellaria dichotoma and a preparation thereof is characterized by comprising the step of obtaining the characteristic spectrum of Stellaria dichotoma and the preparation thereof, wherein the characteristic spectrum comprises 4 common characteristic peaks which are respectively: peak No. 1, peak No. 2, peak No. 3, peak No. 4; taking No. 1 peak dichotomamine B as a reference peak, wherein the relative retention time of the rest characteristic peaks is within +/-10% of a specified value, and the specified values of the relative retention time of the characteristic peaks are respectively as follows:
peak 2 had a relative retention time of 1.24;
peak 3 had a relative retention time of 1.28;
peak No. 4 had a relative retention time of 2.02.
2. The method of building according to claim 1, comprising the steps of,
preparation of a test solution: taking a sample to be detected of starwort root to prepare a test solution;
chromatographic conditions are as follows: performing high performance liquid chromatography with octadecylsilane chemically bonded silica as filler; using acetonitrile as a mobile phase A and acetic acid as a mobile phase B, and performing gradient elution, wherein the gradient elution procedure comprises the following steps: 0-1.5min, mobile phase A: the mobile phase B is 0.7-1.5%: 99.3 to 98.5 percent; 1.5-3.5min, mobile phase A: 1.5-5.0% of mobile phase B: 98.5-95.0%; 3.5-4.0min, mobile phase A: 5.0-10.0% of mobile phase B: 95.0 to 90.0 percent; 4.0-9.0min, mobile phase A: 10.0-13.5% of mobile phase B: 90.0-86.5%; 9.0-10.5min, mobile phase A: 13.5-12.0% of mobile phase B: 86.5 to 88.0 percent; 10.5-13.5min, mobile phase A: the mobile phase B is 12.0-26.0%: 88.0 to 74.0 percent; 13.5-18.5min, mobile phase A: 26.0-43.0% of mobile phase B: 74.0-57.0%; the detection wavelength is 250-290 nm; the flow rate is 0.2-0.7 ml/min;
and (3) determination: and sucking the test solution, injecting into an ultra-high performance liquid chromatograph, and measuring to obtain the test solution.
3. Method of establishing according to claim 1 or 2, characterized in that the chromatographic conditions are: performing high performance liquid chromatography with octadecylsilane chemically bonded silica as filler and detection wavelength of 260-280 nm; the flow rate is 0.3-0.5 ml/min; the column temperature is 29-31 ℃; using acetonitrile as mobile phase A and using volume fraction of 0.01-0.1% acetic acid as mobile phase B, and carrying out gradient elution.
4. The method according to claim 2 or 3, wherein the specific steps of preparing the test solution comprise,
grinding radix Stellariae sample to be tested, precisely weighing, extracting with water for 10-30min, filtering, and collecting the filtrate.
5. The method of any one of claims 2-4, wherein the concentration of the test solution is 40-75 mg/ml.
6. The method of establishing according to claim 4 or 5, characterized in that the method of extraction is an ultrasonic extraction method or a reflux extraction method.
7. The establishing method according to any one of claims 2-6, wherein the Bupleurum falcatum sample to be tested is Bupleurum falcatum medicinal material, Bupleurum falcatum decoction pieces, Bupleurum falcatum standard decoction lyophilized powder or Bupleurum falcatum formula granules.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102329369A (en) * | 2011-05-31 | 2012-01-25 | 苏州派腾生物医药科技有限公司 | Preparation method of dichotomin H |
| WO2012113113A1 (en) * | 2011-02-25 | 2012-08-30 | Mei Chunyong | Herbal composition and use thereof |
| CN102928547A (en) * | 2012-09-07 | 2013-02-13 | 广东省中医研究所 | Quality detection method of starwort root formula granule |
| CN105181625A (en) * | 2015-09-17 | 2015-12-23 | 东莞理工学院 | Method for detecting full-ingredient granule Chinese herbal medicine starwort roots based on terahertz spectrum technology |
-
2020
- 2020-08-26 CN CN202010872945.8A patent/CN113759011B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2012113113A1 (en) * | 2011-02-25 | 2012-08-30 | Mei Chunyong | Herbal composition and use thereof |
| CN102329369A (en) * | 2011-05-31 | 2012-01-25 | 苏州派腾生物医药科技有限公司 | Preparation method of dichotomin H |
| CN102928547A (en) * | 2012-09-07 | 2013-02-13 | 广东省中医研究所 | Quality detection method of starwort root formula granule |
| CN105181625A (en) * | 2015-09-17 | 2015-12-23 | 东莞理工学院 | Method for detecting full-ingredient granule Chinese herbal medicine starwort roots based on terahertz spectrum technology |
Non-Patent Citations (4)
| Title |
|---|
| BOHANG SUN 等: "Structures of New β-Carboline-Type Alkaloids with Antiallergic Effects from Stellaria dichotoma" * |
| XIAOSONG YANG 等: "Study on the Fingerprint of Stellarlae Radix by UPLC" * |
| 刘训红 等: "银柴胡高效液相色谱指纹图谱的初步研究" * |
| 叶方 等: "银柴胡的研究进展" * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114216980A (en) * | 2021-12-14 | 2022-03-22 | 宁夏大学 | Method for establishing HPLC-ELSD (high Performance liquid chromatography-evaporative light scattering) fingerprint spectrum of starwort root |
| CN114216980B (en) * | 2021-12-14 | 2023-12-29 | 宁夏大学 | Method for establishing HPLC-ELSD fingerprint of starwort root |
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