CN113759023A - Preparation process and evaluation method of angelica sinensis Sini decoction - Google Patents

Preparation process and evaluation method of angelica sinensis Sini decoction Download PDF

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CN113759023A
CN113759023A CN202110267096.8A CN202110267096A CN113759023A CN 113759023 A CN113759023 A CN 113759023A CN 202110267096 A CN202110267096 A CN 202110267096A CN 113759023 A CN113759023 A CN 113759023A
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张志强
高晓燕
罗赣
韩妮娜
王颖
梁勇满
付静
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Beijing Tcmages Pharmaceutical Co Ltd
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Abstract

The invention provides a preparation process and an evaluation method of angelica sinensis Sini decoction, wherein the preparation process comprises the following steps: using at least the key quality attribute similarity coefficient S of the characteristic spectrum or fingerprint spectrumcqaEvaluating the process parameters as evaluation indexes; the evaluation process is as follows: s by the characteristic spectrum or fingerprint spectrumcqaScreening out the process parameters corresponding to the characteristic spectrum or the fingerprint spectrum with the maximum value closer to the numerical value 1 as the optimal process parameters; wherein S iscqaThe value is the relative peak area of each characteristic peak in the test sample corresponding to the S peak/the relative peak area of each characteristic peak in the standard decoction corresponding to the S peak. The invention can more simply, conveniently and accurately screen out the process corresponding to the product with more consistent material basis with the clinical standard decoction, and the preparation process is evaluated, so that the process can be effectively ensured to pass throughThe substance reference substance or compound preparation prepared by the process can keep consistent with the substance basis of clinical standard decoction, thereby ensuring the consistency of curative effect.

Description

Preparation process and evaluation method of angelica sinensis Sini decoction
Technical Field
The invention relates to the field of traditional Chinese medicines, and in particular relates to a preparation process and an evaluation method of angelica sinensis Sini decoction.
Background
The angelica Sini decoction comes from Shang Zhong Jing Shang Han Lun of the Han Dynasty, and the decoction method comprises the following steps: three or two angelica, three or two cassia twig (peeled), three or two peony, three or two asarum, two ricepaper pith, two liquorice (roasted), twenty five Chinese dates () are added with eight liters of water, three liters are boiled, dregs are removed, one liter is taken warmly, and three days are taken. The original text records "those with cold hands and feet and thready and faint pulse, and the one with Dang Gui Si reverse decoction. The Dang Gui Siqi Tang is a classic prescription for treating blood deficiency and cold syncope. The formula is a common formula for warming channels and dispelling cold, nourishing blood and dredging collaterals, and is mainly used for treating cold hands and feet, pale tongue with white fur and extremely thready pulse.
In the "material for declaration in the ancient classic famous Chinese medicinal compound preparation material standard" (request for comments), which is published by the national drug administration, the following provisions are explicitly described: except for the molding process, the other preparation methods should be basically consistent with the records of ancient medical books. The classic famous prescription material standard refers to the standard of the traditional Chinese medicine medicinal material prepared according to the ancient classic famous prescription preparation method recorded in the ancient medical books.
However, the existing pharmaceutical research thinking of the angelica sinica rehd soup is not consistent with the clinical practical application, the existing preparation process of the angelica sinica rehd soup is generally guided by the cream yield, the content of effective index components and the maximum value of the transfer rate of the effective index components, the preparation process is researched by the guidance, and the comprehensive comparative research on the specific components of the clinical standard soup is not carried out. Therefore, the material basis of the product obtained by the existing preparation process of the angelica Sini decoction is usually not very consistent with that of the clinical standard decoction, and the consistency of the clinical curative effect cannot be further ensured.
Disclosure of Invention
Therefore, the technical problem to be solved by the invention is to overcome the defect that the preparation process of the angelica Sini decoction in the prior art is usually guided by the cream yield, the content of effective index components and the maximum value of the transfer rate of the effective index components, so that the product is inconsistent with the material basis contained in the clinical standard decoction; therefore, the process evaluation method for screening the product which is more consistent with the material basis contained in the clinical standard decoction can be more simply, conveniently and accurately provided.
An evaluation method of a preparation process of angelica sinensis Sini decoction comprises the following steps:
using at least the key quality attribute similarity coefficient S of the characteristic spectrum or fingerprint spectrumcqaEvaluating the process parameters as evaluation indexes; the evaluation process is as follows: s by the characteristic spectrum or fingerprint spectrumcqaScreening out the process parameters corresponding to the characteristic spectrum or the fingerprint spectrum with the maximum value closer to the numerical value 1 as the optimal process parameters;
wherein, the key quality attribute similarity coefficient S of the characteristic map or the fingerprint mapcqaThe acquisition process comprises the following steps: obtaining standard decoction and test sample of Angelicae Gigantis radix Sini decoction; the standard decoction is an ancient method decoction, and the sample is a substance reference substance or compound preparation of the Angelica sinensis Sini decoction prepared by adopting corresponding process parameters; detecting standard decoction and sample to obtain characteristic spectrum or fingerprint, using one component of the standard decoction as index component, using characteristic peak corresponding to the index component as S peak, obtaining relative peak area of each characteristic peak of the characteristic spectrum or fingerprint of the sample and standard decoction relative to the S peak, calculating S according to the relative peak area of each characteristic peakcqaA value; scqaThe value is the relative peak area of each characteristic peak in the test sample corresponding to the S peak/the relative peak area of each characteristic peak in the standard decoction corresponding to the S peak.
S of characteristic or fingerprintcqaThe number of the values is the same as the number of characteristic peaks in the characteristic spectrum or the fingerprint spectrum, and one characteristic peak corresponds to one ScqaThe value is obtained. In particular, S of a characteristic or fingerprint spectrumcqaIn the characteristics ofS of Peak 1cqaThe value is the relative peak area of characteristic peak 1 corresponding to S peak in the test sample/the relative peak area of characteristic peak 1 corresponding to S peak in the standard decoction. The material reference object in the invention refers to an intermediate obtained in the production process, such as: intermediate preparations such as extract, concentrated solution, extract and dry powder, and the compound preparation refers to finished preparations such as granules and powder containing auxiliary materials.
The detection method of the characteristic spectrum or the fingerprint spectrum is a high performance liquid chromatography, and the chromatographic conditions of the high performance liquid chromatography are as follows:
a chromatographic column: a chromatographic column using octadecylsilane chemically bonded silica as a filler; detection wavelength: 230nm to 280 nm; taking acetonitrile as a mobile phase A, taking a phosphoric acid aqueous solution with the volume concentration of 0.04-0.12% as a mobile phase B, and eluting according to the following gradient elution procedure:
Figure BDA0002970434920000021
in the gradient elution procedure, A11 is 3-8%, A12 is 7-13%, A13 is 10-15%, A14 is 12-17%, A15 is 15-20%, A16 is 17-25%, A17 is 22-35%, A18 is 42-50%, A19 is 75-85%, A20 is 90-98%, A12 < A13 < A14.
In the detection method of the characteristic spectrum or the fingerprint spectrum, the column length of a chromatographic column is 150 mm-250 mm, the inner diameter is 4.6mm, the granularity is 3 mu m-5 mu m, and the column temperature is 28-32 ℃; the flow rate is 0.8ml/min to 1.2 ml/min; the number of theoretical plates is not less than 2000 calculated according to the paeoniflorin peak.
Processing with standard decoction to obtain standard solution, processing with sample to obtain sample solution, and detecting with standard solution and sample solution for characteristic spectrum or fingerprint spectrum;
the standard solution was obtained by the following procedure: taking a proper amount of standard decoction at 3-5 ℃ and 8000 r-min-1～9000r·min-1Centrifuging at high speed for 15-20 min under the condition, taking supernatant at 3-5 ℃ for 12000 r.min-1～13000r·min-1High-speed centrifugation for 15 min-20 m under the conditionin, taking supernatant, filtering, and taking subsequent filtrate to obtain standard solution;
when the sample is an extract or a concentrate, precisely measuring an appropriate amount of the concentrate, and measuring at 3-5 deg.C and 8000-9000 r/min-1Centrifuging at high speed for 15-20 min under the condition, taking supernatant at 3-5 ℃ for 12000 r.min-1～13000r·min-1Centrifuging at high speed for 15-20 min under the condition, taking supernatant, filtering, and taking subsequent filtrate to obtain the final product;
when the test sample is solid, the obtaining process of the test sample solution is as follows: taking a sample, precisely weighing, performing constant volume with purified water, performing ultrasonic treatment until the freeze-dried powder is completely dissolved, taking the solution at 3-5 ℃ and 12000 r.min-1～13000r·min-1Centrifuging at high speed for 15-20 min under the condition, taking supernatant, filtering, and taking subsequent filtrate to obtain the product.
The characteristic spectrum or fingerprint spectrum of the standard decoction has not less than 19 characteristic peaks, and the characteristic peak corresponding to paeoniflorin is used as an S peak;
acquiring the relative retention time of all characteristic peaks in the characteristic spectrum or the fingerprint relative to the S peak, and setting the relative retention time of all characteristic peaks in the characteristic spectrum or the fingerprint of the standard decoction as a specified value, wherein the relative retention time of each characteristic peak in the characteristic spectrum or the fingerprint of the test sample is within +/-8% of the specified value;
the prescribed values include: 0.849, 0.881, 1.000, 1.307, 1.428, 1.472, 1.615, 1.738, 1.785, 1.833, 1.901, 2.027, 2.067, 2.125, 2.194, 2.359, 2.371, 2.399, 2.405;
s of the characteristic spectrum or fingerprint spectrumcqaThe value is in the range of 0.70 to 1.30. Specifically, S, which is closer to the value 1cqaThe value is S in the range of 0.70 to 1.30cqaA value of S in the range of 0.90 to 1.10cqaThe value is obtained.
The evaluation index also comprises at least one of the content of effective index components, the transfer rate of the effective index components, the cream yield, or the similarity of a characteristic spectrum or a fingerprint spectrum;
when the evaluation index is the content of the effective index components, the transfer rate or the cream yield of the effective index components, the evaluation index of the test sample is within +/-30% of the evaluation index corresponding to the standard decoction;
when the evaluation index is the similarity of the characteristic spectrum or the fingerprint spectrum, the similarity range is as follows: 0.900 to 1.000.
The content of the effective index components is detected by adopting a high performance liquid chromatography, and the chromatographic conditions of the high performance liquid chromatography are as follows:
octadecylsilane chemically bonded silica is used as a filler for the chromatographic column; taking acetonitrile as a mobile phase A, taking a 0.04-0.12% phosphoric acid solution as a mobile phase B, and carrying out gradient elution according to the specification in the following table; the detection wavelength is 230 nm-280 nm, and the number of theoretical plates is not less than 2000 calculated according to paeoniflorin;
Figure BDA0002970434920000041
in the gradient elution procedure, A21 is 14-18%, A22 is 45-55%, and A23 is 95-100%.
In the content determination and detection method, the length of a chromatographic column is 150 mm-250 mm, the inner diameter is 4.6mm, the granularity is 3 mu m-5 mu m, and the column temperature is 28-32 ℃; the flow rate is 0.8ml/min to 1.2 ml/min; the number of theoretical plates is not less than 2000 calculated according to the paeoniflorin peak.
When the content of the effective index components is detected by adopting a high performance liquid chromatography, the effective index components in the reference substance solution are as follows: paeoniflorin, ammonium glycyrrhizinate, asaricin, ligustilide, and cinnamaldehyde, wherein the solvent in the reference solution is alcohol solution;
the standard solution was obtained by the following procedure: taking standard decoction at 3-5 ℃ and 12000 r.min-1～13000r·min-1Centrifuging at high speed for 15-20 min under the condition, taking supernatant, filtering, and taking subsequent filtrate to obtain standard solution;
when the sample is an extract or a concentrated solution, precisely measuring the concentrated solution at 3-5 deg.C and 12000 r.min-1～13000r·min-1Centrifuging at high speed for 15-20 min under the condition, and takingFiltering the clear liquid, and taking the subsequent filtrate to obtain the final product;
when the test sample is solid, the obtaining process of the test sample solution is as follows: taking a sample, precisely weighing, performing constant volume with purified water, performing ultrasonic treatment until the freeze-dried powder is completely dissolved, taking the solution at 3-5 ℃ and 12000 r.min-1～13000r·min-1Centrifuging at high speed for 15-20 min under the condition, taking supernatant, filtering, and taking subsequent filtrate to obtain the product.
The cream yield range is 13.78-33.78%, the powder collection rate range is 15.79-21.79%, the paeoniflorin transfer rate range is 32.40-48.50%, the ammonium glycyrrhizinate transfer rate range is 28.41-53.72%, the cinnamic aldehyde transfer rate range is 0.32-3.17%, the ligustilide transfer rate range is 11.53-204.83%, and the asarone transfer rate range is 86.06-181.70%.
The preparation process of the angelica Sini decoction comprises the following steps: adding 20 times of water into the raw materials of the Angelica sinensis Sini decoction, soaking for 30min before decocting, decocting with a cover, boiling with strong fire, decocting with slow fire for 45min, decocting once with 300 meshes, filtering while hot to obtain an extract, concentrating at 60 deg.C for 1-1.5h, and freeze-drying.
The technical scheme of the invention has the following advantages:
1. the invention provides a new quality control index ScqaValue, through ScqaThe value can effectively control whether the material basis contained in the products prepared by different production process steps is consistent with the standard decoction or not, so that the method can be used for evaluating the material basis consistency of the material reference substance or the compound preparation prepared by different preparation processes and the standard decoction, and can screen out the optimal preparation process capable of ensuring the material basis consistency. In particular, can be represented by ScqaComparing the similarity of the intermediate obtained in each step with the standard decoction, wherein the closer the value is to the value 1, the more consistent the key quality attribute index of the intermediate is with the standard decoction, the smaller the material change caused in the process is; thereby effectively promoting the reduction of the difference of the material basis material between the material basis material object or the compound preparation of the meridian preparation angelica sinensis Sini-Rei decoction and the traditional standard decoction, and further avoiding the phenomenon that the meridian preparation angelica sinensis Sini-Rei decoctionThe clinical curative effect of the traditional decoction is greatly different from that of the traditional decoction, so that the consistency of the clinical curative effect is ensured;
at the same time, by the ScqaThe value is set, so that the process corresponding to the material reference substance or the compound preparation which is most similar to the material basis of the traditional standard decoction can be selected more simply, conveniently and efficiently as the optimal process, a more simple, convenient and effective process quality control method for process screening is provided, the analysis time is effectively shortened, and the cost is saved.
2. S in the evaluation method provided by the inventioncqaThe method can be used for screening out the process corresponding to the substance reference substance or compound preparation which is more consistent with the traditional standard decoction and has the closest substance basis, and can also effectively monitor the quality of the substance reference substance or compound preparation, and ensure the stability of the quality of finished products between batches, thereby ensuring the safety and effectiveness of the prepared angelica Sini-Rev decoction substance reference substance, compound preparation and compound preparation to the maximum extent.
3. The standard decoction is prepared according to the method specification recorded by the ancient books, the preparation method is different from that of the Japanese Hanfang standard decoction, the prescription amount of the angelica Sini decoction is determined through the ancient and modern conversion and clinical application documents, and the standard decoction prepared by the invention is more in line with the traditional Chinese medicine theory, can represent the material basis of the decoction described by the ancient books and can reflect the clinical curative effect; in addition, whether the decoction is covered or not, the soaking time, the decoction times, the filtration pore diameter and the like are considered in the extraction process in the previous period, so that the process of process research can be more comprehensively embodied.
4. The evaluation method provided by the invention does not only comprise ScqaThe value also includes the content of effective index components, the transfer rate of the effective index components, the cream yield, the fingerprint spectrum or the similarity of characteristic spectrum, and the like, and the multi-dimensional substance comparison is carried out through the further optimization and limitation of the evaluation indexes, for example, the cream yield range of the sample is limited to 13.78-33.78%, the powder collection rate range is limited to 15.79-21.79%, the transfer rate range of paeoniflorin is 32.40-48.50%, the transfer rate range of ammonium glycyrrhizinate is 28.41-53.72%, the transfer rate range of cinnamaldehyde is 0.32-3.17%, and the transfer rate of ligusticum is 0.32-3.17%The transfer rate range of the lactone is 11.53-204.83%, and the transfer rate range of the asarone is 86.06-181.70%; within the parameter condition range, the material basis of the intermediate or the finished product prepared by the preparation process is better ensured to be more consistent with that of the standard decoction, and the consistency of clinical curative effect is better ensured.
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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 shows the fingerprint of the standard decoction of the present invention.
FIG. 2 is S of the concentrates prepared at different concentration temperatures according to the present inventioncqaThe resulting line graph.
FIG. 3 is S of the concentrates prepared at different concentration times in the present inventioncqaThe resulting line graph.
FIG. 4 shows S of dry powders prepared by different drying methods in the present inventioncqaThe resulting line graph.
Detailed Description
Example 1
The present example performs stable research and determination of standard decoction process, and also performs investigation of influence of concentration step on material basis in angelica sinensis sini decoction, specifically as follows:
1. research on process stability of standard decoction
1.1 chromatographic conditions
1.1.1 high performance liquid fingerprint/characteristic spectrum method of SINITANG decoction
Chromatographic conditions and system adaptability are realized by taking octadecylsilane chemically bonded silica as a filler (the column length is 250mm, the inner diameter is 4.6mm, and the particle size is 5 mu m) as a chromatographic column; detection wavelength: 230 nm; acetonitrile is taken as a mobile phase A, a phosphoric acid solution with the volume concentration of 0.05 percent is taken as a mobile phase B, and the column temperature is 30 ℃; the flow rate was 1.0ml/min, and elution was carried out according to the gradient elution procedure shown in Table 1 below, and the number of theoretical plates was not less than 2000 calculated from the paeoniflorin peak.
TABLE 1 gradient elution Table
Figure BDA0002970434920000061
Figure BDA0002970434920000071
Preparation of control solutions: taking appropriate amount of ferulic acid, paeoniflorin, liquiritin, cinnamaldehyde, ammonium glycyrrhizinate, and asaricin reference, precisely weighing, and adding methanol to obtain a solution containing 20, 34.3, 23.6, 20, 68.8, and 19.1 μ g/ml of the reference per 1ml-1And (4) mixing the solution to obtain the finished product.
Preparation of standard solution: collecting appropriate amount of standard decoction at 4 deg.C and 9000r min-1Centrifuging at high speed for 20min under the condition, collecting 1mL of supernatant at 4 deg.C and 13000r min-1Centrifuging at high speed for 20min, collecting supernatant, filtering, and collecting filtrate to obtain standard solution.
Preparation of a test solution:
when the sample is extractive solution, precisely measuring appropriate amount of extractive solution at 4 deg.C and 9000 r/min-1Centrifuging at high speed for 20min under the condition, collecting 1mL of supernatant at 4 deg.C and 13000r min-1Centrifuging at high speed for 20min, collecting supernatant, filtering, and collecting filtrate.
When the sample is concentrated solution, precisely measuring appropriate amount of concentrated solution, and processing at 4 deg.C and 9000 r/min-1Centrifuging at high speed for 20min under the condition, collecting 1mL of supernatant at 4 deg.C and 13000r min-1Centrifuging at high speed for 20min, collecting supernatant, filtering, and collecting filtrate.
When the test sample is freeze-dried powder, taking about 0.2g of the test sample, precisely weighing, placing in a 10mL volumetric flask, carrying out volume fixing by deionized water, carrying out ultrasonic treatment for 3-5 min until the freeze-dried powder is completely dissolved, and taking the above1mL of the solution was dissolved at 4 ℃ and 13000 r.min-1Centrifuging at high speed for 20min, collecting supernatant, filtering, and collecting filtrate.
The determination method comprises precisely sucking 10 μ l of each of the reference solution and the sample solution, injecting into liquid chromatograph, and determining.
19 characteristic peaks are presented in the characteristic map of the test sample, wherein the retention time of the 19 characteristic peaks is the same as that of the corresponding characteristic peak of the reference substance, the peak corresponding to the paeoniflorin reference substance is an S peak, the relative retention time of each characteristic peak and the S peak is calculated, and the relative retention time is within +/-8% of a specified value. The specified values are: 0.849 (peak 1), 0.881 (peak 2), 1.000 (peak 3), 1.307 (peak 4), 1.428 (peak 5), 1.472 (peak 6), 1.615 (peak 7), 1.738 (peak 8), 1.785 (peak 9), 1.833 (peak 10), 1.901 (peak 11), 2.027 (peak 12), 2.067 (peak 13), 2.125 (peak 14), 2.194 (peak 15), 2.359 (peak 16), 2.371 (peak 17), 2.399 (peak 18), 2.405 (peak 19). Wherein, peak 3: paeoniflorin, peak 4: liquiritin, peak 10: cinnamaldehyde, peak 11: ammonium glycyrrhizinate, peak 17: asarone; peak 3 is the S peak.
ScqaThe value: controlling the relative retention time of 19 characteristic peaks calculated in the characteristic spectrum or fingerprint spectrum of the test sample to be within +/-8% of a specified value; among the 19 characteristic peaks, S of characteristic spectrum or fingerprint spectrumcqaAt least 14 of the values are in the range of 0.70 to 1.30.
1.1.2 method for determining standard content of Angelica sinensis Sini decoction
Chromatographic conditions and system applicability test with octadecylsilane chemically bonded silica as filler (column length 250mm, inner diameter 4.6mm, particle size 5 μm); acetonitrile is used as a mobile phase A, 0.05 percent phosphoric acid solution is used as a mobile phase B, and the column temperature is 30 ℃; the flow rate was 1.0ml/min, and gradient elution was performed as specified in the following table; the detection wavelength was 280 nm. The number of theoretical plates is not less than 2000 calculated by paeoniflorin.
TABLE 2 gradient elution Table
Figure BDA0002970434920000081
Preparation of control solutions: paeoniflorin, ammonium glycyrrhizinate, asafetida, ligustilide and cinnamaldehyde are used as reference substances, and methanol is adopted to prepare into a concentration of 20 μ g-1、30.5μg.ml-1、30μg.ml-1、23μg.ml-1、30μg.ml-1The control solution of (4).
Preparation of standard solution: collecting appropriate amount of standard decoction, and processing at 4 deg.C and 13000 r.min-1Centrifuging at high speed for 20min, collecting supernatant, filtering, and collecting filtrate to obtain standard solution
Preparation of a test solution:
when the sample is extractive solution, precisely measuring the extractive solution at 4 deg.C and 13000 r.min-1Centrifuging at high speed for 20min, collecting supernatant, filtering, and collecting filtrate to obtain standard solution.
Precisely measuring appropriate amount of concentrated solution when the sample is concentrated solution, centrifuging the decoction at 4 deg.C and 13000r min-1 for 20min, collecting supernatant, filtering, and collecting filtrate.
When the test sample is lyophilized powder, taking about 0.5g of the test sample, precisely weighing, placing in a 10mL volumetric flask, fixing the volume with deionized water, performing ultrasonic treatment for 3-5 min until the lyophilized powder is completely dissolved, taking 1mL of the above solution, dissolving at 4 ℃, 13000 r.min-1Centrifuging at high speed for 20min, filtering, and collecting the filtrate.
The determination method comprises precisely sucking 10 μ l of each of the reference solution and the sample solution, injecting into liquid chromatograph, and determining.
1.2 Process stability confirmation of Standard decoction
Weighing 3 parts of prescription dosage of decoction pieces of angelica sinensis Sini decoction with the same batch number according to the prescription proportion, wherein the prescription dosage is as follows: angelica sinensis three and two, cassia twig three and two (peeled), peony three and two, asarum herb three and two, ricepaper pith two, licorice root two (roasted), and Chinese date twenty-five (). Adding 1600ml of water in parallel, covering and soaking for 30min, setting control parameters, starting equipment, boiling with strong fire (500W), keeping with slow fire (300W) for 45min, decocting once, filtering with 200-mesh pharmacopoeia sieve while hot, rapidly cooling to room temperature, measuring the volume of the liquid medicine, wherein the volume range is as follows: 540ml to 660ml, and refrigerating the filtrate for later use. Taking the original liquid medicine to perform index component content and fingerprint detection according to the chromatographic conditions in the step 1.1, calculating the cream yield, the transfer rate and the similarity of the fingerprint, and the detection results are shown in tables 3-6.
Table 3 Process verification of the cream yield of three batches of extract
Figure BDA0002970434920000091
Table 4 Process verification of index component content of three batches of extract
Figure BDA0002970434920000092
Table 5 Process verification three batches of extract relative retention time
Figure BDA0002970434920000093
Table 6 Process verification three batches of extract fingerprint similarity comparison
Figure BDA0002970434920000101
The reference fingerprint refers to the fingerprint generated by the three batches of extracting solutions for extraction and verification through fingerprint similarity software. In summary, the transfer rates of paeoniflorin, glycyrrhizic acid, cinnamaldehyde, ligustilide and asarone verified by 3 batches of decoction and filtration processes are all within the specified range of the standard decoction of angelica Sini decoction; the 3-batch decocting and filtering process verifies that the paste rate is also in the specified range of the standard angelica Sini decoction; in 3 batches of decocting and filtering processes, the components of the decoction are consistent with the components of the standard decoction of the angelica Sini decoction by contrasting the fingerprint, which shows that the types of the substances are consistent in the extraction and filtering stages, thereby proving that the decocting and filtering process is stable and feasible.
2. Influence of concentration step on material basis in Angelica sinensis Sini decoction
2.1 examination of concentration temperature
And (4) decocting by adopting the determined optimal process parameters to obtain an extracting solution, wherein the extracting solution is a standard decoction. Obtaining a concentrated solution: collecting 1200ml extractive solution, dividing into four parts, and concentrating under reduced pressure at four temperatures of 50 deg.C, 60 deg.C, 70 deg.C and 80 deg.C to 100ml respectively to obtain radix Angelicae sinensis Sini decoction concentrate.
Preparing the concentrated solution into sample solution, preparing standard decoction of the same batch into standard solution, detecting fingerprint and transfer rate of paeoniflorin, glycyrrhizic acid, cinnamaldehyde, ligustilide, and asaricin, and calculating to obtain S of each concentrated solutioncqaThe value is obtained. The results of the measurements are shown in FIGS. 1 to 2 and tables 7 to 9. Wherein, figure 1 is the fingerprint of the standard decoction.
TABLE 7 comparison of transfer rates of effective index components at different concentration temperatures
Figure BDA0002970434920000102
TABLE 8 comparison of fingerprint similarity for different concentration temperatures
Figure BDA0002970434920000103
Figure BDA0002970434920000111
TABLE 9 analysis of different concentration temperature fingerprint results
Figure BDA0002970434920000112
The contrast fingerprint spectrum refers to the fingerprint spectrum of the standard decoction of accompanying contrast, and the following results show that: the transfer rates of paeoniflorin, glycyrrhizic acid, cinnamaldehyde, ligustilide and asarone are less than 10% when the concentration temperature is 50 deg.C, 60 deg.C, 70 deg.C and 80 deg.C, the concentration temperature is 50 deg.C, 60 deg.C, 70 deg.C,The similarity between the fingerprint at 80 deg.C and standard decoction of Angelicae Gigantis radix SINITANG decoction is greater than 0.9, which indicates that the species of the concentrated solution is identical to that of the standard decoction, and the fingerprint S is examined at concentration temperaturecqaThe results of the values and the line graphs show that the number of 19 characteristic peaks in the range of 0.70-1.30 in the samples at 50 ℃, 70 ℃ and 80 ℃ is less than 14, the number of peaks in the range of 0.70-1.30 in the samples at 60 ℃ is more than 14, and the concentration temperature of the tentative angelica sinica decoction is 60 ℃ in comprehensive consideration.
2.2 investigation of the concentration time
Extracting according to the selected optimal extraction conditions, collecting 600ml extractive solution of the other batch of Chinese medicinal decoction pieces, and concentrating under reduced pressure at 60 deg.C for 1.5 hr and 3 hr respectively to obtain radix Angelicae sinensis Sini decoction concentrate. Taking the extractive solution of the same batch as standard decoction, preparing into standard solution, diluting the concentrated solution, detecting the transfer rate and fingerprint of paeoniflorin, glycyrrhizic acid, cinnamaldehyde, ligustilide, and asaricin, and calculating to obtain S of each concentrated solutioncqaThe values, the results of the measurements are shown in FIG. 3 and tables 10-12.
TABLE 10 comparison of transfer rates of effective index components at different concentration times
Figure BDA0002970434920000121
TABLE 11 analysis of fingerprint results for different concentration times
Figure BDA0002970434920000122
Figure BDA0002970434920000131
TABLE 12 comparison of fingerprint similarity at different concentration times
Figure BDA0002970434920000132
The control fingerprint is fingerprint of standard decoction of accompanying control, and the result shows that, concentration time is 1.5 hr and 3.0 hr, transfer rates RSD of paeoniflorin, glycyrrhizic acid, cinnamaldehyde, ligustilide and asaricin are all less than 3.0%, and the similarity between fingerprint of concentration time is 1.5 hr and 3.0 hr and fingerprint of standard decoction of Angelica sinensis Sini-Ding decoction is more than 0.9, which indicates that the species of substances in the concentrated solution is consistent with that of standard decoction, and the concentration time examines fingerprint ScqaThe results of the values and the line graphs show that 19 characteristic peaks of the sample of 1.5h are all within the range of 0.70-1.30, less than 14 characteristic peaks of the sample of 3.0h are within the range of 0.70-1.30, and the concentration time of the tentative angelica sinica decoction is 1.5h in comprehensive consideration.
2.3 three batches of verifications
Weighing the standard decoction of the angelica Sini decoction of the same batch number, and preparing three batches of concentrated solution according to the concentration temperature of 60 ℃ and the concentration time of 1.5 h. Taking the concentrated solution to perform index component content and fingerprint detection, and calculating cream yield, transfer rate and ScqaThe values and the measurement results are shown in tables 13 to 15.
Table 13 Process verification of index component transfer rates of three batches of concentrates
Figure BDA0002970434920000133
Figure BDA0002970434920000141
TABLE 14 Process verification three batches of concentrate fingerprint similarity comparison
Figure BDA0002970434920000142
TABLE 15 analysis of three lots of concentration verification fingerprint results
Figure BDA0002970434920000143
Figure BDA0002970434920000151
The comparison fingerprint spectrum refers to the fingerprint spectrum of the standard decoction of accompanying comparison, and the results in the tables 14 to 16 show that: the transfer rates of paeoniflorin, glycyrrhizic acid, cinnamaldehyde, ligustilide and asafetida verified by 3 batches of three-batch concentration process are all within the specified range of standard decoction of angelica sinica decoction; the paste yield of the 3 batches of the three batches of concentration verification processes is also within the specified range of the standard decoction of the angelica Sini decoction; the components of the angelica sinensis Sini decoction are verified to be consistent with the components of the reference fingerprint spectrum of the standard decoction of the angelica sinensis Sini decoction by 3 batches of concentration processes; fingerprint spectrum S verified by three batches of concentrationcqaThe results show that 19 characteristic peaks in the concentrated sample are all in the range of 0.70-1.30, which indicates that the substance types are consistent with those of the standard decoction in the concentration stage. In conclusion: the concentration process of the invention is stable and feasible.
2.4 summary
The concentration process is determined by the above investigation of the factors such as concentration temperature, volume, time and the like as follows: the concentration temperature is 60 ℃, and the concentration time is 1.5 h.
Example 2
The present example performs the investigation of the influence of the drying step on the material basis in the angelica sini decoction, and the specific investigation process is as follows:
1.1 examination of different drying modes
Extracting and concentrating another batch of Chinese medicinal decoction pieces by the optimal process in example 1, and drying 3 parts of the obtained concentrated solution in 60 deg.C electrothermal blowing drying oven, reduced pressure drying oven, and vacuum freeze dryer respectively to obtain final product. The obtained dry powder is subjected to index component content detection and fingerprint spectrum detection. The detection method and conditions were the same as in example 1. Detecting the content and S of the effective index componentcqaThe results and fingerprint similarity are shown in tables 16-18 and figure 4.
TABLE 16 examination of effective index component transfer rate in drying mode
Figure BDA0002970434920000152
Figure BDA0002970434920000161
TABLE 17 comparison of fingerprint similarity for drying
Figure BDA0002970434920000162
TABLE 18 analysis of finger-print results for different drying modes
Figure BDA0002970434920000163
The control fingerprint is fingerprint of standard decoction of accompanying control, and the result shows that reduced pressure drying, atmospheric pressure drying, and freeze drying, except glycyrrhizic acid, transfer rates RSD of paeoniflorin, cinnamaldehyde, ligustilide, and asaricin are all less than 3.0%, similarity of normal pressure and reduced pressure drying characteristic patterns and standard decoction characteristic patterns in different drying modes is less than 0.900, similarity of freeze drying and standard decoction characteristic patterns is greater than 0.900, and S of normal pressure and reduced pressure dryingcqaThe trend of the value line graph is inconsistent with that of vacuum freeze drying, and S of 19 characteristic peaks is freeze driedcqaThe number of peaks with the value of 0.70-1.30 is more than or equal to 14, and the S with the characteristic peak less than 14 is obtained by drying under reduced pressure and normal pressurecqaThe value is in the range of 0.70 to 1.30. Comprehensively considering, the tentative angelica Sini decoction drying mode is freeze drying.
1.2 verification of the optimal drying regime for three batches
Taking the concentrated solution of the same batch used in the drying, preparing three batches of dry powder by adopting the optimal drying mode, and performing content detection and fingerprint similarity detection on index components by adopting the dry powder as a test sample, wherein the detection results are shown in tables 19-20.
Table 19 Process verification of index component transfer rates of three batches of lyophilized powder
Figure BDA0002970434920000171
Table 20 Process verification three-batch lyophilized powder fingerprint similarity comparison
Figure BDA0002970434920000172
The comparison fingerprint spectrum refers to a fingerprint spectrum generated by the three batches of freeze-dried powder through fingerprint spectrum similarity software. The above results show that: the transfer rates of paeoniflorin, glycyrrhizic acid, cinnamaldehyde, ligustilide and asafetida verified by 3 batches of three-batch freeze drying process are all within the specified range of standard decoction of angelica sinica decoction; the cream yield of the 3 batches of the three batches of the freeze drying verification process is also in the specified range of the standard decoction of the angelica Sini decoction; the components of the freeze-drying process of 3 batches are verified to be consistent with the components of the standard decoction of the angelica Sini decoction by contrasting the fingerprint, which shows that the substance types are consistent with the standard decoction in the drying stage, and the freeze-drying process is stable and feasible.
1.3 summary
The optimal drying process is determined by the investigation of different drying modes as follows: and (5) vacuum freeze drying.
Example 3
The embodiment provides a method for detecting the standard material object of the angelica sinica rehd prepared under different preparation process conditions, including examples 1-6, and according to the detection result, the evaluation of the angelica sinica rehd preparation process can be simply and effectively realized, so that the optimal preparation process conditions are screened out.
Wherein, S of the characteristic spectrum or fingerprint spectrumcqaThe detection method and the detection method of the content of the effective index components are the same as the method described in example 1, and the detection is carried out by adopting a high performance liquid chromatography fingerprint detection method, and the detection conditions and implementation are the sameExample 1 is also identical except that the test article is different.
Example 1: weighing angelica Sini decoction pieces according to the proportion of the prescription, adding 1600ml of water, soaking for 1 hour, decocting until the liquid medicine amount is 540ml, filtering the liquid medicine, concentrating at 60 ℃, concentrating for 1.5 hours, and drying under normal pressure to obtain an angelica Sini decoction substance reference sample 1.
The detection result of the sample 1 is as follows: the cream yield is 33.77%, the powder recovery rate is 20.89%, the paeoniflorin transfer rate is 37.80%, the ammonium glycyrrhizinate transfer rate is 40.98%, the cinnamaldehyde transfer rate is 2.15%, the ligustilide transfer rate is 90.11%, the asarone transfer rate is 98.67%, and the similarity coefficient of a high performance liquid chromatography fingerprint is 0.980; s of sample 1cqaThe values are given in table 21 below.
Example 2: weighing angelica Sini decoction pieces according to the proportion of the prescription, adding 1600ml of water, soaking for 0.5 hour, decocting until the liquid medicine amount is 600ml, filtering the liquid medicine, concentrating at 60 ℃, concentrating for 1.5 hours, and drying under reduced pressure to obtain an angelica Sini decoction substance reference sample 2.
The detection result of sample 2 is: the cream yield is 31.89%, the powder recovery rate is 20.79%, the paeoniflorin transfer rate is 40.87%, the ammonium glycyrrhizinate transfer rate is 50.35%, the cinnamaldehyde transfer rate is 2.56%, the ligustilide transfer rate is 87.35%, the asarone transfer rate is 89.45%, and the similarity coefficient of a high performance liquid fingerprint is 0.975; s of sample 2cqaThe values are given in table 21 below.
Example 3: weighing angelica Sini decoction pieces according to the proportion of the prescription, adding 1600ml of water, not soaking, decocting until the liquid medicine amount is 660ml, filtering the liquid medicine, concentrating at 50 ℃, concentrating for 1.0h, freeze-drying at the pre-freezing temperature of-27 ℃, wherein the freeze-drying time is 35h, and obtaining the angelica Sini decoction reference sample 3.
The detection result of sample 3 is: the cream yield is 29.09%, the powder recovery rate is 25.79%, the paeoniflorin transfer rate is 42.87%, the ammonium glycyrrhizinate transfer rate is 49.05%, the cinnamaldehyde transfer rate is 3.09%, the ligustilide transfer rate is 88.78%, the asarone transfer rate is 87.42%, and the high performance liquid fingerprint similarity coefficient is 0.964; s of sample 3cqaThe values are given in table 21 below.
Example 4: weighing angelica Sini decoction pieces according to the proportion of the prescription, adding 1600ml of water, soaking for 0.5 h, decocting until the liquid medicine amount is 620ml, filtering, concentrating the filtered liquid medicine at 80 ℃, concentrating for 1.0h, and freeze-drying at the pre-freezing temperature of-25 ℃ for 37h to obtain an angelica Sini decoction material reference sample 4.
The detection result of sample 4 is: the cream yield is 32.90%, the powder recovery rate is 20.69%, the paeoniflorin transfer rate is 42.89%, the ammonium glycyrrhizinate transfer rate is 54.89%, the cinnamaldehyde transfer rate is 3.16%, the ligustilide transfer rate is 98.56%, the asarone transfer rate is 99.67%, and the high performance liquid fingerprint similarity coefficient is 0.967; s of sample 4cqaThe values are given in table 21 below.
Example 5: weighing angelica Sini decoction pieces according to the proportion of the prescription, adding 1600ml of water, soaking for 1.0 hour, decocting until the liquid medicine amount is 600ml, filtering, concentrating the filtered liquid medicine at 70 ℃, concentrating for 2 hours, and freeze-drying at the pre-freezing temperature of-25 ℃ for 39 hours to obtain an angelica Sini decoction reference sample 5.
The detection result of the sample 5 is as follows: the cream yield is 25.89%, the powder recovery rate is 19.67%, the paeoniflorin transfer rate is 39.87%, the ammonium glycyrrhizinate transfer rate is 53.35%, the cinnamaldehyde transfer rate is 3.11%, the ligustilide transfer rate is 90.56%, the asarone transfer rate is 90.00%, and the high performance liquid fingerprint similarity coefficient is 0.987; s of sample 5cqaThe values are given in table 21 below.
Example 6: weighing angelica Sini decoction pieces according to the proportion of the prescription, adding 1600ml of water, soaking for 1.0 hour, decocting until the liquid medicine amount is 600ml, filtering, concentrating the filtered liquid medicine at 60 ℃, concentrating for 1.5 hours, and freeze-drying at the pre-freezing temperature of-27 ℃ for 35 hours to obtain an angelica Sini decoction material reference sample 6.
The detection result of sample 6 is: the cream yield is 28.54%, the powder recovery rate is 20.04%, the paeoniflorin transfer rate is 40.15%, the ammonium glycyrrhizinate transfer rate is 51.45%, the cinnamaldehyde transfer rate is 3.22%, the ligustilide transfer rate is 89.48%, the asarone transfer rate is 91.04%, and the high performance liquid fingerprint similarity coefficient is 0.975; s of sample 6cqaThe values are given in table 21 below.
The above samples 1 to 6 were tested by the test method described in example 1, and the test results are shown in table 21 below.
TABLE 21
Figure BDA0002970434920000191
Figure BDA0002970434920000201
From the above table, it can be seen that: s of 19 chromatographic peaks in samples 1-5cqaThe number of peaks having a value within a predetermined range of 0.70 to 1.30 is less than 14, and S of 19 chromatographic peaks in sample 6cqaThe number of peaks having a value within a predetermined range of 0.70 to 1.30 is > 14.
In conclusion: the invention can evaluate the consistency of the preparation process of the examples 1-6 and the key quality attribute of the standard decoction, and the display result shows that: the number of 19 peaks in the sample 6 within the range of 0.7-1.3 is more than 14, and the quality control requirement is met. Therefore, the preferred preparation processes of example 6 are all optimized processes.
Example 4
The difference between this example and example 1 is that the chromatographic conditions for detection are different, and the specific settings are as follows:
the following characteristic spectrum or fingerprint detection is respectively carried out by using the substance reference material object of the angelica sinica decoction of two batches in example 6, and the detection method is as follows:
detecting one,
The detection method of the characteristic spectrum or the fingerprint spectrum is a high performance liquid chromatography, and the chromatographic conditions of the high performance liquid chromatography are as follows:
a chromatographic column: a chromatographic column using octadecylsilane chemically bonded silica as a filler; detection wavelength: 280 nm; acetonitrile was used as the mobile phase A, and a 0.04% by volume phosphoric acid aqueous solution was used as the mobile phase B, and the column temperature was 28 ℃, the flow rate was 1.2ml/min, the column length was 150mm, the inner diameter was 4.6 μm, and the particle size was 3.0 μm, and elution was carried out according to the gradient elution procedure shown in Table 22 below: the number of theoretical plates is not less than 2000 calculated according to paeoniflorin;
TABLE 22
Figure BDA0002970434920000202
Figure BDA0002970434920000211
The process of obtaining the test solution comprises the following steps: taking a sample, precisely weighing, adding purified water to constant volume, ultrasonic treating until the lyophilized powder is completely dissolved, taking the above solution at 5 deg.C and 12500 r.min-1Centrifuging at high speed for 18min under the condition, collecting supernatant, filtering, and collecting filtrate to obtain sample solution for detecting characteristic spectrum or fingerprint of sample 1.
The content of the effective index components is detected by adopting a high performance liquid chromatography, and the chromatographic conditions are as follows:
a chromatographic column: octadecylsilane chemically bonded silica is used as a filling agent; acetonitrile is taken as a mobile phase A, 0.06 percent phosphoric acid solution is taken as a mobile phase B, the detection wavelength is 280nm, the column temperature is 29 ℃, and the flow rate is 1.0 ml/min; the column length of 250mm, the inner diameter of 4.6 μm, the particle size of 5.0 μm, according to the following table 23 in the specification of gradient elution; the number of theoretical plates is not less than 2000 calculated according to paeoniflorin;
TABLE 23
Figure BDA0002970434920000212
The process of obtaining the test solution comprises the following steps: taking a sample, precisely weighing, adding purified water to constant volume, ultrasonic treating until the lyophilized powder is completely dissolved, collecting the above solution at 3 deg.C and 13000 r.min-1Centrifuging at high speed for 20min under the condition, collecting supernatant, filtering, and collecting filtrate to obtain sample solution for detecting effective index component of sample 1.
The detection result of the sample 1 is as follows: the cream yield is 28.35%, the paeoniflorin transfer rate is 38.23%, the ammonium glycyrrhetate transfer rate is 50.11%, the cinnamaldehyde transfer rate is 3.10%, the ligustilide transfer rate is 89.20%, the asarone transfer rate is 90.11%, and the high performance liquid fingerprint similarity coefficient is 0.982; s of sample 1cqaThe values are given in Table 26 below.
Detecting II,
The detection method of the characteristic spectrum or the fingerprint spectrum is a high performance liquid chromatography, and the chromatographic conditions of the high performance liquid chromatography are as follows:
a chromatographic column: a chromatographic column using octadecylsilane chemically bonded silica as a filler; detection wavelength: 270 nm; acetonitrile is taken as a mobile phase A, phosphoric acid aqueous solution with the volume concentration of 0.1 percent is taken as a mobile phase B, the column temperature is 32 ℃, and the flow rate is 0.8 ml/min; : the column length is 250mm, the inner diameter is 4.6 μm, the particle size is 5.0 μm, the elution is carried out according to the gradient elution program shown in the following table 24, and the number of theoretical plates is not less than 2000 calculated according to paeoniflorin;
watch 24
Figure BDA0002970434920000221
The process of obtaining the test solution comprises the following steps: taking a sample, precisely weighing, adding purified water to desired volume, ultrasonic treating to dissolve the lyophilized powder completely, collecting the above solution at 4 deg.C and 12000 r.min-1Centrifuging at high speed for 15min under the condition, collecting supernatant, filtering, and collecting filtrate to obtain sample solution for detecting characteristic spectrum or fingerprint of sample 2.
The content of the effective index components is detected by adopting a high performance liquid chromatography, and the chromatographic conditions are as follows:
a chromatographic column: octadecylsilane chemically bonded silica is used as a filling agent; acetonitrile is taken as a mobile phase A, 0.05 percent phosphoric acid solution is taken as a mobile phase B, the detection wavelength is 270nm, the column temperature is 28 ℃, and the flow rate is 1.2 ml/min; the column length was 250mm, the inner diameter was 4.6 μm, and the particle size was 5.0 μm, and gradient elution was carried out as specified in Table 25 below; the number of theoretical plates is not less than 2000 calculated according to paeoniflorin;
TABLE 25
Figure BDA0002970434920000222
The process of obtaining the test solution comprises the following steps: taking a test sample, precisely weighing, performing constant volume with purified water, performing ultrasonic treatment until the freeze-dried powder is completely dissolved,collecting the above solution at 3 deg.C and 12000r min-1Centrifuging at high speed for 15min under the condition, collecting supernatant, filtering, and collecting filtrate to obtain sample solution for detecting effective index component of sample 2.
The detection results of sample 2 were: the cream yield is 28.19%, the paeoniflorin transfer rate is 36.90%, the ammonium glycyrrhetate transfer rate is 52.09%, the cinnamaldehyde transfer rate is 3.11%, the ligustilide transfer rate is 89.35%, the asarone transfer rate is 91.91%, and the high performance liquid phase fingerprint similarity coefficient is 0.992; s of sample 2cqaThe values are given in Table 26 below.
Watch 26
Figure BDA0002970434920000231
The samples 1 and 2 are the same as the standard sample of the angelica sinensis Sini-Rev soup substance of example 6 in example 3, and the comparison of the detection results with those of example 6 shows that: s obtained by respectively detecting two batches of angelica Sini soup substance reference substancescqaThe difference between the value and the result of example 6 in Table 21 was within 5%, and the difference between the cream yield and the effective index component transfer rate of the two batches of the Angelica sinensis Sini decoction reference substance and the result of example 6 was within 5%. The detection conditions of the above detection method of the present invention are also proved to be applicable.
Example 5
In the embodiment, a compound obtained by compounding different batches of traditional Chinese medicine decoction pieces meeting requirements is adopted, at least 20 batches of extracting solution are prepared by adopting an ancient decoction preparation method, the paste yield, the powder collection rate and the transfer rate of effective index components of the extracting solution are detected, and the detection results are as follows:
the cream yield range is 13.78-33.78%, the powder collection rate range is 15.79-21.79%, the paeoniflorin transfer rate range is 32.40-48.50%, the ammonium glycyrrhizinate transfer rate range is 28.41-53.72%, the cinnamic aldehyde transfer rate range is 0.32-3.17%, the ligustilide transfer rate range is 11.53-204.83%, and the asarone transfer rate range is 86.06-181.70%.
From this, it is understood that when the satellite extract is used as the standard solution, the process parameters can be further evaluated using the evaluation indexes.
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 therefrom are within the scope of the invention.

Claims (10)

1. An evaluation method of a preparation process of angelica Sini decoction is characterized by comprising the following steps: using at least the key quality attribute similarity coefficient S of the characteristic spectrum or fingerprint spectrumcqaEvaluating the process parameters as evaluation indexes; the evaluation process is as follows: s by the characteristic spectrum or fingerprint spectrumcqaScreening out the process parameters corresponding to the characteristic spectrum or the fingerprint spectrum with the maximum value closer to the numerical value 1 as the optimal process parameters;
wherein, the key quality attribute similarity coefficient S of the characteristic map or the fingerprint mapcqaThe acquisition process comprises the following steps: obtaining standard decoction and test sample of Angelicae Gigantis radix Sini decoction; the standard decoction is an ancient method decoction, and the sample is a substance reference substance or compound preparation of the Angelica sinensis Sini decoction prepared by adopting corresponding process parameters; detecting standard decoction and sample to obtain characteristic spectrum or fingerprint, using one component of the standard decoction as index component, using characteristic peak corresponding to the index component as S peak, obtaining relative peak area of each characteristic peak of the characteristic spectrum or fingerprint of the sample and standard decoction relative to the S peak, calculating S according to the relative peak area of each characteristic peakcqaA value; scqaThe value is the relative peak area of each characteristic peak in the test sample corresponding to the S peak/the relative peak area of each characteristic peak in the standard decoction corresponding to the S peak.
2. The method for evaluating the preparation process of the angelica sinica decoction according to claim 1, wherein the detection method of the characteristic spectrum or the fingerprint spectrum is a high performance liquid chromatography, and the chromatographic conditions of the high performance liquid chromatography are as follows:
a chromatographic column: a chromatographic column using octadecylsilane chemically bonded silica as a filler; detection wavelength: 230nm to 280 nm; taking acetonitrile as a mobile phase A, taking a phosphoric acid aqueous solution with the volume concentration of 0.04-0.12% as a mobile phase B, and eluting according to the following gradient elution procedure:
Figure FDA0002970434910000011
in the gradient elution procedure, A11 is 3-8%, A12 is 7-13%, A13 is 10-15%, A14 is 12-17%, A15 is 15-20%, A16 is 17-25%, A17 is 22-35%, A18 is 42-50%, A19 is 75-85%, A20 is 90-98%, A12 < A13 < A14.
3. The method for evaluating the process for preparing angelica Sini decoction according to claim 2, wherein the step of preparing the angelica Sini decoction comprises the steps of,
in the detection method of the characteristic spectrum or the fingerprint spectrum, the column length of a chromatographic column is 150 mm-250 mm, the inner diameter is 4.6mm, the granularity is 3 mu m-5 mu m, and the column temperature is 28-32 ℃; the flow rate is 0.8ml/min to 1.2 ml/min; the number of theoretical plates is not less than 2000 calculated according to the paeoniflorin peak.
4. The method for evaluating the preparation process of the angelica sinica exsiccata according to claim 2 or 3, wherein the standard decoction is treated to obtain a standard solution, the test sample is treated to obtain a test sample solution, and the standard solution and the test sample solution are used for detecting a characteristic spectrum or a fingerprint;
the standard solution was obtained by the following procedure: taking standard decoction at 3-5 deg.C and 8000r min-1～9000r·min-1Centrifuging at high speed for 15-20 min under the condition, taking supernatant at 3-5 ℃ for 12000 r.min-1～13000r·min-1Centrifuging at high speed for 15-20 min under the condition, taking supernatant, filtering, taking continuous filteringObtaining a standard solution;
when the sample is an extract or a concentrated solution, precisely measuring an appropriate amount of the concentrated solution at 3-5 deg.C and 8000r min-1～9000r·min-1Centrifuging at high speed for 15-20 min under the condition, taking supernatant at 3-5 ℃ for 12000 r.min-1～13000r·min-1Centrifuging at high speed for 15-20 min under the condition, taking supernatant, filtering, and taking subsequent filtrate to obtain the final product;
when the test sample is solid, the obtaining process of the test sample solution is as follows: taking a sample, precisely weighing, performing constant volume with purified water, performing ultrasonic treatment until the freeze-dried powder is completely dissolved, taking the solution at 3-5 ℃ and 12000 r.min-1～13000r·min-1Centrifuging at high speed for 15-20 min under the condition, taking supernatant, filtering, and taking subsequent filtrate to obtain the product.
5. The method for evaluating the preparation process of angelica Sini decoction according to any one of claims 2 to 4, wherein the standard decoction has not less than 19 characteristic peaks in the characteristic spectrum or fingerprint, and the characteristic peak corresponding to paeoniflorin is used as an S peak; s of the characteristic spectrum or fingerprint spectrumcqaAt least 14 of the values are in the range of 0.70-1.30;
preferably, the relative retention time of all characteristic peaks in the characteristic spectrum or the fingerprint spectrum relative to the S peak is obtained, and if the relative retention time of all characteristic peaks in the characteristic spectrum or the fingerprint spectrum of the standard decoction is a specified value, the relative retention time of each characteristic peak in the characteristic spectrum or the fingerprint spectrum of the test sample is within +/-8% of the specified value;
the prescribed values include: 0.849,0.881,1.000,1.307,1.428,1.472,1.615,1.738,1.785,1.833,1.901,2.027,2.067,2.125,2.194,2.359,2.371,2.399,2.405.
6. The method for evaluating the preparation process of the angelica Sini decoction according to any one of claims 1 to 5, wherein the evaluation index further comprises at least one of the content of effective index components, the transfer rate of the effective index components, the cream yield, or the similarity of a characteristic spectrum or a fingerprint spectrum;
when the evaluation index is the content of the effective index components, the transfer rate or the cream yield of the effective index components, the evaluation index of the test sample is within +/-30% of the evaluation index corresponding to the standard decoction;
when the evaluation index is the similarity of the characteristic spectrum or the fingerprint spectrum, the similarity range is as follows: 0.900 to 1.000.
7. The method for evaluating the preparation process of angelica sinica rehd as claimed in claim 6, wherein the content of the effective index components is detected by high performance liquid chromatography, and the chromatographic conditions of the high performance liquid chromatography are as follows:
octadecylsilane chemically bonded silica is used as a filler for the chromatographic column; acetonitrile is taken as a mobile phase A, 0.04-0.1% phosphoric acid solution is taken as a mobile phase B, and gradient elution is carried out according to the specification in the following table; the detection wavelength is 230 nm-280 nm, and the number of theoretical plates is not less than 2000 calculated according to paeoniflorin;
Figure FDA0002970434910000031
in the gradient elution procedure, A11 is 14-18%, A12 is 45-55%, A13 is 95-100%, A11 is more than A12 is more than A13;
in the content determination and detection method, the length of a chromatographic column is 150 mm-250 mm, the inner diameter is 4.6mm, the granularity is 3 mu m-5 mu m, and the column temperature is 28-32 ℃; the flow rate is 0.8ml/min to 1.2 ml/min; the number of theoretical plates is not less than 2000 calculated according to the paeoniflorin peak.
8. The method for evaluating the process for preparing angelica sinica rehd according to claim 7, wherein when the content of the effective index components is detected by a high performance liquid chromatography, the effective index components in the reference solution are as follows: paeoniflorin, ammonium glycyrrhizinate, asaricin, ligustilide, and cinnamaldehyde, wherein the solvent in the reference solution is alcohol solution;
the standard solution was obtained by the following procedure: taking standard decoction at 3-5 ℃ and 12000 r.min-1～13000r·min-1Centrifuging at high speed for 15-20 min under the condition, taking supernatant, filtering, and taking subsequent filtrate to obtain standard solution;
when the sample is an extract or a concentrated solution, precisely measuring the concentrated solution at 3-5 deg.C and 12000 r.min-1～13000r·min-1Centrifuging at high speed for 15-20 min under the condition, taking supernatant, filtering, and taking subsequent filtrate to obtain the final product;
when the test sample is solid, the obtaining process of the test sample solution is as follows: taking a sample, precisely weighing, diluting with deionized water to desired volume, ultrasonic treating to dissolve the lyophilized powder, and dissolving the solution at 3-5 deg.C and 12000 r.min-1～13000r·min-1Centrifuging at high speed for 5-20 min under the condition, taking supernatant, filtering, and taking subsequent filtrate to obtain the product.
9. The method for evaluating the process for preparing Dang Gui Sini Tang according to claim 6, wherein the step of preparing the Dang Gui Sini Tang is carried out,
the cream yield range is 13.78-33.78%, the powder collection rate range is 15.79-21.79%, the paeoniflorin transfer rate range is 32.40-48.50%, the ammonium glycyrrhizinate transfer rate range is 28.41-53.72%, the cinnamic aldehyde transfer rate range is 0.32-3.17%, the ligustilide transfer rate range is 11.53-204.83%, and the asarone transfer rate range is 86.06-181.70%.
10. The process for preparing angelica sinica rehd soup evaluated by the evaluation method according to any one of claims 1 to 9, comprising: soaking radix Angelicae sinensis Sini decoction in 1600ml water for 30min before decocting, decocting with cover, boiling with strong fire, decocting with slow fire for 45min, decocting once, filtering with 200 mesh filter to obtain extractive solution, concentrating at 60 deg.C for 1-1.5 hr, and freeze drying.
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